The crayfish plague is a devastating disease of European freshwater crayfish and is caused by the oomycete Aphanomyces astaci (Ap. astaci), which was introduced to Europe around 1860. All European species of freshwater crayfish are susceptible to the disease, including the white-clawed crayfish Austropotamobius pallipes. The aquatic mould is primarily spread by North American crayfish species and can also disperse rapidly through contaminated wet gear moved between water bodies. This spread, coupled with competition from non-indigenous crayfish, has drastically reduced and fragmented native crayfish populations across Europe. Remarkably, the island of Ireland remained free from the crayfish plague pathogen for over 100 years, providing a refuge for A. pallipes. However, this changed in 1987 when a mass mortality event was linked to the pathogen marking the pathogen's introduction to the region. Fortunately, the plague did not establish and was not detected again in Ireland until 2015 when a molecular analysis linked a mass mortality event in the Erne catchment to Ap. astaci. Since then, the pathogen has appeared across the island. Between 2015 and 2023, Ap. astaci was detected in 18 Irish water catchments, revealing multiple genotypes. Intriguingly, the pathogen in Ireland exists without its natural host species. The uneven distribution of various genetic lineages strongly suggests human-mediated transport of zoospores via contaminated water equipment as a primary cause of spread. This review details the timeline of these events, Ap. astaci’s introductions into Ireland, its rapid spread, the genotypes documented, the issue of non-indigenous crayfish species in Ireland and management efforts.

Ureaplasma species (Ureaplasma spp.) are commonly found as commensals in the human urogenital tracts , but their overgrowth can lead to infection in the urogenital tract and at distal sites. Treatment of Ureaplasma spp. infections is difficult. The lack of a cell wall confers resistance to all β-lactam and glycopeptide antibiotics whereas the lack of de novo synthesis of folic acid makes them resistant to sulfonamides and diaminopyrimidines. Thus, there is a need to search for new drugs effective against these bacteria. In the presented studies, the effect of N,N-diethyldithiocarbamate (DDC) at a concentration of 2 µg/ml, 20 µg/ml and 200 µg/ml on the growth and survival of Ureaplasma urealyticum and Ureaplasma parvum was tested. Our results indicated that all DDC concentrations used showed both bacteriostatic and bactericidal activity against both tested strains In brief, for the first time we showed the bacteriostatic and bactericidal activity of DDC against Ureplasma urealyticum and Ureaplasma parvum. Based on literature data, we conclude that this DDC activity is related to the inhibition of urease activity, which is an indispensable enzyme for these bacteria. Further research is needed and the results obtained in the present study should be verified in in vivo models of infection.

O26 enteropathogenic Escherichia coli (EPEC), possesses a unique pseudocapsule composed of polysaccharides identical to the O antigen in their lipopolysaccharide (LPS) membrane. Therefore, the impact of O26 polysaccharides on the immune evasion mechanisms of capsulated O26 EPEC was investigated. Our findings reveal that the concentration of GlcNAc, an immunodominant unit of O26 polysaccharides capable of activating the MBL (Mannose Biding Lecting) pathway of the complement system, was found to be lower in capsulated EPEC O26 compared to naked shiga toxin-producing E. coli (STEC) O26 strain. It was noted that, unlike the naked STEC strain, the capsulated EPEC O26 strain was protected against lysis by the alternative pathway of the com-plement system by inhibiting C3b adhesion to its bacterial surface. Capsulated EPEC O26 strains were also able to avoid phagocytosis by macrophages. However, the immune evasion provided by the capsule was overcome in the presence of anti-O26 polysaccharide antibodies. Additionally, these antibodies were able to recognize O26 EPEC in the presence of biofilm and inhibit their adhesion to human epithelial cells. Overall, the results indicate that the O26 polysaccharides can generate an effective humoral immune response, making them promising antigens for the de-velopment of a vaccine against capsulated O26 E. coli

Thanks to several Vitis vinifera backcrosses with an initial V. vinifera L. × V. rotundifolia (previously Muscadinia rotundifolia) interspecific cross, the MrRUN1/MrRPV1 locus (resistance to downy and powdery mildews) was introgressed in genotypes phenotypically close to V. vinifera varieties. To check the consequences of introgressing parts of the V. rotundifolia genome on gene expression during fruit development, we conducted a comparative RNA-seq study on single berries from different V. vinifera cultivars and V. vinifera × V. rotundifolia hybrids, including ‘G5’ and two derivative microvine lines, ‘MV102’ (resistant) and ‘MV32’ (susceptible) segregating for the MrRUN1/RPV1 locus. RNA-Seq profiles were analyzed on a comprehensive set of single berries from the end of the herbaceous plateau to the ripe stage. Pair-end reads were aligned both on V. vinifera PN40024.v4 reference genome, V. rotundifolia cv ‘Trayshed’ and cv ‘Carlos’, and to the few resistance genes from the original V. rotundifolia cv ‘52’ parent available at NCBI. Weighted Gene Co-expression Network Analysis (WGCNA) led to classifying the differentially expressed genes into 15 modules either preferentially correlated with resistance or berry phenology and composition. Resistance positively correlated transcripts predominantly mapped on the 4-5 Mb distal region of V. rotundifolia chromosome 12 beginning with the MrRUN1/MrRPV1 locus, while the negatively correlated ones mapped on the orthologous V. vinifera region, showing this large extremity of LG12 remained recalcitrant to internal recombination during the successive backcrosses. Some constitutively expressed V. rotundifolia genes were also observed at lower density outside this region. Genes overexpressed in developing berries from resistant accessions, either introgressed from V. rotundifolia, or triggered by these in the vinifera genome, spanned various functional groups, encompassing calcium signal transduction, hormone signaling, transcription factors, plant–pathogen-associated interactions, disease resistance proteins, ROS and phenylpropanoid biosynthesis. This transcriptomic insight provides a foundation for understanding the disease resistance inherent in these hybrid cultivars and suggests a constitutive expression of NIR NBS LRR triggering calcium signaling. Moreover, these results illustrate the magnitude of transcriptomic changes caused by the introgressed V. rotundifolia background in backcrossed hybrids, on a large number of functions largely exceeding the ones constitutively expressed in single resistant gene transformants.

Ethanolic extracts of propolis collected from Apis mellifera hives have been shown to reduce Nosema ceranae infection in honey bee workers. This study aimed to determine the propolis concentration that optimizes the control of N. ceranae infection in honey bee workers. Newly emerged worker bees were individually fed 2 μL of 50% sucrose solution (w/v) containing 105 N. ceranae spores per bee, and treated with 0%, 25%, 50%, and 75% ethanolic extracted propolis in 50% sucrose solution (v/v). The treated bees were maintained in cages at 34 ± 2˚C and 55 ± 5% RH throughout the study. The number of dead bees was counted and removed from the cages daily for 30 days to generate survival curves. The number of N. ceranae spores per bee (infectivity) was quantified using a hemocytometer, and the number of infected bees per hundred bees (infection rate) was determined. The survival rates of N. ceranae infected bees fed 25%, 50%, and 75% propolis (NP1, NP2, NP3, respectively) were significantly higher than those of infected bees not fed propolis (NO). Furthermore, the infectivity and infection rates of NP2 and NP3 groups were significantly lower than those of the NO group. The infectivity and infection rates of the NP1 group were not different than those of the NO group. Our data suggest that 50% and 75% propolis extracts are appropriate concentrations for treating N. ceranae infection in honey bees. Ethanolic extracted propolis can be an alternative treatment for N. ceranae infection in Western honey bees.

This study developed Makgeolli (Korean traditional alcoholic beverage) by adding raw 20% Codium fragile (Cf), a green seaweed, to a mixed base consisting of rice and oats (1:1 ratio) (COM) and analyzed its quality characteristics and antioxidant activity. The rice based Makgeolli supplemented with raw 20% Cf was designated as control (CRM). The physicochemical (pH, acidity), microbiological (yeast, lactic acid bacteria), antioxidant (DPPH, ABTS), and sensory properties (7-henodonic scale) of CRM and COM were investigated during 7 days of fermentation. The pH/acidity levels of CRM and COM reached 3.66/0.68 and 3.69/0.95 after 7 days of fermentation, respectively. Compared to CRM, COM had lower alcohol and sugar contents (P < 0.05) (CRM: 14.5%/9.8 Brix, COM: 11.2%/7.4 Brix). However, COM contained more yeast and lactic acid bacteria (P < 0.05) (CRM: 1.30/4.28 log, COM: 6.04/6.09 log) compared to CRM. The antioxidant activity of DPPH/ABTS (P < 0.05) in COM was approximately 1.5 times that of CRM (CRM: 40.00/51.57%, COM: 56.76/70.91%). In addition, COM had an excellent sensory preference score (P < 0.05) due to its overall fresh flavor and savory taste (CRM: 4.93, COM: 6.06). Therefore, COM was shown to be Makgeolli with enhanced antioxidant function, with yeast and lactic acid bacteria, and improved flavor, suggesting the possibility of commercializing this Makgeolli.

Megaselia scalaris and Senotainia tricuspis are two parasitoid flies of the honeybee Apis mellifera and are known to infest apiaries of different European and Mediterranean countries. However, their prevalence is still little known and no systematic control of their presence and countermeasures are currently adopted. To achieve a better knowledge about their distribution in apiaries, quantitative PCR (qPCR) based methods for their rapid detection directly in hive matrices were developed in this study. These were applied to samples of adult honeybees and hive debris collected in the summer season in Central Italy and Kosovo. In total 16 samples per type per region were analyzed. Both flies were detected in the two geographical contexts but more frequently in Central Italy. The positive apiaries in Central Italy were re-sampled one year later by capturing viable honeybees and isolating emerging flies after differentiation into adults. These were genotyped by sequencing regions of the mitochondrial cytochrome oxidase subunit I gene COI for M. scalaris and cytochrome B gene cytB for S. tricuspis amplified with specific primers designed in this study. Different genotypes of M. scalaris were detected and these were related to biotypes from distant nations. Some fly isolates were shown to harbor DNA or RNA of honeybee microbial pathogens. Results suggested to implement efficient control against these parasites in apiaries to limit their detrimental effects. The qPCR tests that have been developed can facilitate larger screening studies and early detection.

Bacterial infection of the lower respiratory tract frequently occurs in mechanically ventilated pa-tients and may develop into life-threatening conditions. Yet existing diagnostic methods have moderate sensitivity and specificity, which results in the overuse of broad-spectrum antibiotics administrated prophylactically. This study aims to evaluate the suitability of volatile bacterial me-tabolites for breath-based test diagnosing Ventilation-Associated Pneumonia (VAP). The in vitro experiments with pathogenic microorganisms most prevalent in VAP patients (i.e., Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa) were performed to identify bacteria-derived metabolites using specially designed cultivation system enabling headspace sam-pling for GC-MS analysis. Thirty-nine compounds were found to be significantly metabolized by tested species and, therefore, selected for monitoring in the exhaled breath of critically ill, me-chanically ventilated (MV) patients. The emission of volatiles from medical respiratory devices was investigated to estimate the risk of spoiling breath results with exogenous pollutants. Bacterial metabolites were then evaluated to differentiate VAP patients from non-infected MV controls using receiver operating characteristic (ROC) analysis, with AUC, sensitivity, and specificity calculated. Nine bacterial metabolites that passed verification through a non-parametric ANOVA test for significance and LASSO penalization were identified as key discriminators between VAP and non-VAP patients. The diagnostic model achieved an AUC of 0.893, with sensitivity and specificity values of 87% and 82.4%, respectively, being competitive with traditional methods. Further vali-dation could solidify its clinical utility in critical care settings.

Finding, safe and reliable alternatives to fungicides is one of the most actual challenges in agriculture. In this regard, our experiment investigated the effectiveness of different elicitors, botanical extracts and essential oils to control grey mould (Botrytis cinerea) and powdery mildew (Podosphaera aphanis) on strawberry plants. Furthermore, the defence mechanisms activated in plants after treatments were investigated. The trial was conducted in field conditions under plastic tunnel with strawberry plants ‘Elsanta’. In the first experiment, a group of strawberry plants was treated before flowering with elicitors [acibenzolar-S-Methyl – (BTH), chitosan], botanical extracts (seaweed extract, alfalfa hydrolysate) and essential oils (thyme and juniper) to evaluate their ability to reduce the incidence of grey mould infection on flowers. In the second experiment, another group of plants was treated before and after experimental inoculation with P. aphanis. Results indicated that the incidence of flower infected by B. cinerea was significantly reduced by thyme and juniper essential oils, showing values that were not significantly different from the commercial fungicide penconazole (positive control). As a consequence, final yield of essential oil-treated plants was +27% higher as compared to control. No significant differences emerged for other tested products against grey mould. However, gene expression analysis showed an up-regulation of FaEDS1, FaLOX and PR gene expression (FaPR1, FaPR5, FaPR10) in leaves treated with BTH. The other tested natural substances also elicited defence-related genes, however at a lower level than BTH. In the experiment, all treatments applied prior inoculation significantly reduced the incidence and severity of powdery mildew as compared to control. At 28 days after inoculation, chitosan and thyme essential oil applications performed similarly to their positive controls (BTH and penconazole, respectively), showing a significant reduction of disease incidence (by -84 and -92%) as compared to control. Post-inoculum application of essential oils showed similar effective action to fungicide (penconazole) in reducing incidence and severity of powdery mildew. These results indicate that the tested substances could be used as alternatives to fungicides for the control of grey mould and powdery mildew in strawberry, therefore representing a valuable tool for the control of these fungal diseases under the framework of the sustainable agriculture.

The production of recombinant proteins in Escherichia coli is often challenged by cytoplasmic expression due to proteolytic degradation and inclusion body formation. Extracellular expression can overcome these problems by simplifying downstream processing and improving protein yields. This study aims to compare the efficiency of two Bacillus subtilis chitosanase signal peptides in mediating extracellular secretion in E. coli. We identified a naturally occurring mutant signal peptide (mCsn2-SP) from B. subtilis CH2 chitosanase (CH2CSN), which is characterized by a deletion of six amino acids in the N-region relative to the signal peptide (Csn1-SP) from B. subtilis CH1 chitosanase (CH1CSN). The CH1CSN and CH2CSN genes were cloned into the pET-11a vector and protein secretion was evaluated in E. coli BL21(DE3) host cells. Expression was induced with 0.1 mM and 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) at 30°C for one and three days. CH2CSN showed higher secretion levels compared to CH1CSN under all experimental conditions, especially with 0.1 mM IPTG induction for 3 days, which resulted in a 2.37-fold increase in secretion. Furthermore, it was demonstrated that mCsn2-SP is capable of secreting human Cu,Zn-superoxide dismutase (hSOD) in E. coli BL21(DE3) and successfully translocating it to the periplasmic region. This study represents the inaugural investigation into the utilisation of a naturally modified signal peptide, thereby corroborating the assertion that signal peptide deletion variants can influence protein secretion efficiency. Furthermore, the findings substantiate the proposition that such variants can serve as a viable alternative for the secretion of heterologous proteins in E. coli.

Biofilm-associated infections caused by Candida species represent a significant clinical challenge due to their inherent resistance to antifungal therapies. This study evaluates the antifungal efficacy of trans-Cinnamaldehyde, a natural compound derived from Cinnamomum cassia, in comparison to the widely used antifungal agent Nystatin, against both planktonic and biofilm forms of Candida albicans, Candida glabrata, and Candida tropicalis. Biofilms were established in 96-well polystyrene plates and treated with various concentrations of trans-Cinnamaldehyde and Nystatin. The Minimum Biofilm Inhibitory Concentration (MBIC) and planktonic minimum inhibitory concentration (PMIC) were determined using the MTT assay and Scanning Electron Microscopy (SEM) was employed to assess structural changes in biofilms. The results demonstrated that Nystatin exhibited superior antifungal efficacy across all tested species, particularly in biofilm inhibition, with MBIC50 values as low as 0.0015 mg/mL for C. albicans and C. glabrata. In contrast, trans-Cinnamaldehyde showed moderate antifungal activity with MBIC50 values of 0.16 mg/mL for C. albicans and C. glabrata and higher resistance observed in C. tropicalis (MBIC50 of 0.32 mg/mL). SEM analysis revealed significant biofilm disruption in C. albicans following exposure to trans-Cinnamaldehyde, but C. tropicalis exhibited more resilient biofilm structures. Overall, while Nystatin remains the gold standard for antifungal treatment, trans-Cinnamaldehyde presents potential as an adjunctive treatment, particularly for biofilm-related infections in C. albicans and C. glabrata. Further research is required to explore synergistic effects between trans-Cinnamaldehyde and conventional antifungal agents and to optimize its therapeutic application in clinical settings. This study reused measurement results data from our previous publication to include in tables and charts.

Euler sums are alternating (or level two) extension of multiple zeta values (MZVs). Kaneko and Tsumura initiated the study of multiple T-values (MTVs), another level two generalization, by restricting the summation indices in the definition of MZVs to a fixed parity pattern. In this paper, we shall study finite MTVs and their alternating versions which are level two and level four variations of finite MZVs, respectively. We conjecture that all finite MZVs are in the Q-span of finite MTVs which in turn apparently lie in the span of finite Euler sums, and the inclusions are both proper. We shall first provide some structural results for Euler sums of small weights, guided by the author's previous conjecture that the finite Euler sum space of weight w is isomorphic to a quotient Euler sum space of weight w. Then, by utilizing some well-known properties of the classical alternating MTVs, we shall derive a few important Q-linear relations among the finite alternating MTVs, including the reversal, linear shuffle and sum relations. We then compute the upper bound for the dimension of the Q-span of weight w finite (alternating) MTVs for w<9, both rigorously using the newly discovered relations and numerically aided by computers.

Bacterial wilt (BW), caused by Ralstonia solanacearum, is one of the devastating diseases of tomatoes (Solanum lycopersicum L.). The use of resistant cultivars and breeding for genetic resistance is the most effective, economical, and environmentally friendly management strategy for this disease. It is necessary to screen diverse germplasm and cultivated genotypes to identify resistant resources and to develop resistant cultivars in tomatoes to combat the changing pathogen isolates. This study evaluated 40 United States Department of Agriculture (USDA) tomato accessions for their BW resistance to R. solanacearum isolate P822 under greenhouse conditions. The tomato plants were inoculated and visually assessed to observe symptoms and disease severity was scored on a scale of 0 to 4 (0 = no leaf wilted, 1 = 25% of leaves wilted, 2 = 50% leaves wilted, 3 = 75% of leaves wilted, 4 = 100% leaves wilted). Five accessions PI 645370, PI 647306, PI 600993, PI 355110, and PI 270210 were observed as BW resistance with PI 645370 showing the greatest resistance. The broad-sense heritability was estimated as 59.9% based on a 0-4 scale of disease incidence and 42.8% based on the disease severity index for BW resistance. Two distinct clusters (sub-populations) were detected among 39 of the 40 accessions, consisting of 3 and 2 BW resistant accessions in each cluster, respectively, suggesting differences in the genetic base in the five resistance accessions. The resistant accessions will contribute significantly to the tomato breeding program to develop new cultivars with BW resistance.

Tomato (Solanum lycopersicum L.) has almost 190 million tonnes produced in 2023. Tomato fruit suffer losses of up to 50% during harvest and transport, which causes financial hardship, reduces the amount of food available and causes environmental harm. Calcium plays an important role in cell wall strength. This work assessed the use of a calcium transport stimulants (MCAS) to increase the firmness of tomato fruit in the laboratory and the foliar application to tomato plants. Then handling damage was simulated. In the laboratory, the calcium with MCAS significantly increased the fruit firmness compared to the untreated and calcium without MCAS, which were not significantly different. When calcium with MCAS was applied to tomato plants foliarly before harvest, the calcium with MCAS treated fruit were significantly firmer than the untreated or calcium without MCAS treated fruit for up to 10 weeks after harvest, and this was achieved by applying only 0.91 kg ha-1 calcium. Finally, when fruit were exposed to a simulated drop, the calcium with MCAS treated fruit remained firmer than the undamaged fruit or dropped fruit with calcium without MCAS. Calcium with MCAS could significantly increase the fruit firmness and increase the shelf-life of tomatoes while applying less calcium.

Tomato is considered the most important vegetable crop worldwide. Improving the nutritional value of fruits must be based on sustainable production in terms of varieties and fertilization management. This study aimed to improve the nutritional value (total soluble solids, acidity, lycopene, β-carotene, polyphenols, macro and microelements) of two tomato varieties (‘Cristal’ and ‘Siriana’) under three fertilization types (NPK chemical fertilizer, chicken manure and biological fertilizer with microorganisms) for the greenhouse. Primary metabolism compounds do not vary significantly according to the type of fertilizer used. The results for the antioxidant compounds showed a better effect of biological fertilization compared to chemical fertilizer and control unfertilized. The tomato fruits from the local cultivar (‘Siriana’) are richer in nutritional compounds such as rutin, regardless of the type of fertilization, which denotes a good ability to adapt to crop conditions. Tomato cultivars reacted positively to microbiological fertilization compare to chemical, thus producing nutritious fruits under sustainable management. Tomato fruits were richer in the quality of microelement contents.

Background/Objectives: The Perilla frutescens plant has historically been used to protect against inflammation and redox stress. This has been attributed in part to the high content of various polyphenols. Despite this, the complement of polyphenolic components in Perilla extract is not yet fully defined. The aim of this study was to characterise the polyphenolic composition in Perilla extract, and its effect on the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), which during inflammation and oxidative stress binds to the antioxidant response element (ARE) found on the promoter of phase II antioxidant enzymes. Methods: Following extraction using four solvents of different properties, the total phenolic, flavonoid and ortho-diphenolic content of Perilla extract were determined. Individual phenolic compounds were identified and quantified by RP-HPLC. Dual Luciferase assay using a reporter plasmid containing a human NQO1 ARE-luciferase fusion was employed to determine the potential protective effect of the Perilla fractions against redox stress. Results: HPLC analysis revealed that the highest phenolic content was present in the polar extracts. A total of 35 distinct polyphenolic compounds were identified, with rosmarinic acid being the major constituent. On cells in vitro, the methanol Perilla fraction showed protection against redox stress, with the luciferase assay displaying up to 1.5 fold increase in the induction of the human NQO1 ARE-luciferase reporter. Conclusions: While Perilla extract seems to be mainly protecting against inflammation and redox stress through the action of rosmarinic acid, further work is required on the synergystic effects between different polyphenols.

This study aimed to find alternative feeds or supplementary feeds that can perform similarly to the existing commercial feeds thereby reducing farmer expense and cutting down their cost of production and increasing their profitability. The formulations for the study included four grow-out treatments mainly: control (commercial feed), taro+control (powdered Colocasia esculenta+commercial fish feed), bloodmeal (formulated fish feed based on bloodmeal), banana stem+control (chopped banana stem+commercial fish feed). A total of 12,000 fingerlings initially weighing 16 g (TL=10 cm) individually were assigned randomly to the four treatments (3000 in each cage measuring 6 x 6 m). The result of the study showed no significant differences between feed treatments from 6^th week onwards, whether with weight [MS=0.130, df=3, F=1.14, P=0.333] or total length of the treatments [MS=0.008, df=3, F=0.40, P=0.75]. The FCR ratio showed bloodmeal (1.60) as lowest, followed by the control (1.65), and taro+commercial (1.71) and then banana stem+commercial (2.18). In terms of weight gain, fish from the bloodmeal treatment gained the highest (236.97 g), followed by control (223.33 g), then taro (217.67 g) and banana stem (196.99 g). As a result, we find that any of the tested treatments can serve as an alternative feed to using purely commercial fish feeds. Cost-benefit analysis showed that a better gross profit margin of 42% and cost-benefit ratio of 1.37 was observed in taro+commercial feed diet.

Developing recombinant Saccharomyces cerevisiae strains capable of transporting and fermenting cellobiose directly is a promising strategy for second-generation ethanol production from lignocellulosic biomass. In this study we cloned and expressed in the CEN.PK2-1C strain an intracellular βglucosidase (SpBGL7) from Spathaspora passalidarum, and co-expressed the cellobiose transporter SiHXT2.4 from Scheffersomyces illinoinensis, and two putative transporters from Candida tropicalis (CtCBT1) and Meyerozyma guilliermondii (MgCBT2). While all three transporters allowed cell growth on cellobiose, only the MgCBT2 permease allowed cellobiose fermentation, although cellobiose consumption was stuck (incomplete). The analysis of the βglucosidase and transport activities revealed that the cells stopped to consume cellobiose due to a drop in the transport activity. Since ubiquitinylation of lysine residues at the N- or C-terminal domains of the permease are involved in the endocytosis and degradation of sugar transporters, we constructed truncated versions of the permease lacking lysine residues at the C-terminal domain (MgCBT2ΔC), and at both the C- and N-terminal domain (MgCBT2ΔNΔC), and co-expressed these permeases with the SpBGL7 βglucosidase in an industrial strain. While the strain harboring the MgCBT2ΔC transporter continued to produce stuck cellobiose fermentations as the wild-type MgCBT2 permease, the strain with the MgCBT2ΔNΔC permease was able to consume and ferment all the cellobiose present in the medium. Thus, our results highlight the importance of expressing cellobiose transporters lacking lysine at the N- and C-terminal domains for efficient cellobiose fermentation by recombinant S. cerevisiae.

We studied yeast hand carriage of 260 students from medicine (78), nursing (89), and medical laboratory scientist (93). We isolated yeasts in 27 students (10.4%), with no differences between careers or gender. A significant lower prevalence of carriage was shown in the clinical cycle (2.7%) compared to the basic cycle (13.5%) (p=0.022) and the preclinical cycle (13.5%) (p=0.014).Increased handwashing frequency and alcohol gel use decreased yeast carriage. Students who applied moisturizing hand cream two or more times a day had a lower frequency of yeast carriage (3.4%) compared to those who did not use it or used it only once a day (16.5%), showing a significant difference (p=0.016). The most prevalent species was C. parapsilosis sensu stricto (81.5%), followed by Meyerozyma guilliermondii (C. guillermondii) (7.4%), Trichosporon mucoides (7.4%), and R. mucilagenosa (3.7%). And one case showed mixed carriage of C. parapsilosis and C. albicans. All strains were sensitive to voriconazole, caspofungin, and anidulafungin. This study shows the yeast carriage on the hands of health students, especially of C. parapsilosis, and its reduction with the frequent use of infection control measures and moisturizing hand cream.

Amphibians are widely known as a prolific source of bioactive metabolites. In this work we isolated and characterized compounds with antiparasitic activity from the oocytes of the toad Rhinella alata collected in Panama. Bio-guided isolation and structural elucidation was carried out using chromatographic and spectroscopic techniques. The organic crude extract was subjected to solid phase extraction followed by HPLC purification of the fraction with in vitro activity against Trypanosoma cruzi trypomastigotes. Seven steroids (1-7) of the bufadienolide family were isolated and their structures determined using NMR and MS analyses; of these 19-formyl-dyscinobufotalin (3) is reported as a new natural product. Compounds 1, 3-7, resulted with a good anti-trypanosomal activity profile, among these 16β-hydroxyl-hellebrigenin (1) and bufalin (7) showed significant selectivity for T. cruzi. Furthermore, molecular docking analysis showed compounds 1, 3 and 7 interact through H-bonds with the amino acid residues GLN-19, ASP-158, HIS-159 and TRP-177 from cruzipain, at the catalytic site. Given the lack of therapeutic options to currently treat the American trypanosomiasis, this work can serve as the basis for further studies that aim the development of bufadienolides or their derivatives as drugs against Chagas disease.

Cytokinesis, the process by which the cytoplasm divides to generate two daughter cells after mitosis, is a crucial stage of the cell cycle. Successful cytokinesis must be coordinated with chromosome segregation and requires the fine orchestration of several processes, such as constriction of the actomyosin ring, membrane reorganisation, and, in fungi, cell wall deposition. In Saccharomyces cerevisiae, commonly known as budding yeast, septins play a pivotal role in the control of cytokinesis by assisting the assembly of the cytokinetic machinery at the division site and controlling its activity. Yeast septins form a collar at the division site that undergoes major dynamic transitions during the cell cycle. This review will discuss the functions of septins in yeast cytokinesis, their regulation and the implications of their dynamic remodelling for cell division.

Glucosinolates, the main secondary metabolites accumulated in cruciferous flora, have a major impact on fortifying plant immunity against diverse pathogens. Although Botrytis cinerea exhibits varying sensitivity to these compounds, current research has yet to fully understand the intricate mechanisms governing its response to glucosinolates. Different species of the genus Botrytis were exposed to glucosinolate-breakdown products revealing that B. fabae, B. deweyae and B. convoluta, species lacking the mfsG transporter gene (Bcin06g00026), were more sensitive to glucosin-olate-breakdown products than Botrytis species containing that gene such as B. cinerea, B. pseudo-cinerea and B. byssoidea. This finding was further corroborated by the inability of species lacking the mfsG gene to infect plants with a high concentration of glucosinolate breakdown products. These results challenge established correlations, revealing varying aggressiveness on different plant substrates. Expression analysis highlighted the gene's induction in the presence of isothiocyanate, and bioinformatic investigation identified homologous genes in other Botrytis species. Our study underscored the importance of advanced biotechnology to help understand these proteins and thus offer innovative solutions for agriculture

Leishmania infantum is an hemopathogen of importance for the health of domestic dogs (Canis lupus familiaris), causing canine leishmaniasis (CanL), as well as it is the etiological agent of human visceral leishmaniasis (HVL). This parasite was not reported in southern Brazil until the early 2000s, but CanL and HVL were increasingly reported in the last 15 years, mainly in cities bordering Argentina. The present study aimed to detect L. infantum in urban dogs and to determine the main clinical manifestations in the infected animals from Uruguaiana, a city with a high incidence of CanL. Fifty-one dogs suspected of having CanL in the urban perimeter of the city were clinically examined by veterinarians and investigated for the occurrence of L. infantum with two immunoassays (rapid chromatography test and ELISA) and real-time PCR. Clinical signs were compared in positive and negative L. infantum animals. A total of 31 dogs (60.8%) were infected with L. infantum. The main clinical manifestations associated to CanL dogs were onychogryphosis and peeling (p&lt;0.05). L. infantum was frequently detected in urban dogs from Uruguaiana, highlighting the health concerning situation in this city. The occurrence of some clinical signs (onychogryphosis/peeling) could help to detect more frequently CanL in the canine population.

This paper aimed to evaluate the growth, gas exchange and Chl a fluorescence in ipê-rosa seedlings, subjected to levels of 100, 75, 50 and 25% of water replacement (LWR), in a greenhouse. The morphometric variables of plant height, diameter at stem height, number of leaves and leaflets, root length and volume, plant dry mass and leaf area were evaluated. The potential evapotranspiration of seedlings (ETc) was obtained using direct weighing, considering as a reference the water replacement of 100% of the mass variation between subsequent days; the cultivation coefficients (kc) were obtained by the ratio between ETc and the reference evapotranspiration (ETo) obtained by the Penman-Montheit FAO-56 method. Biomass and evapotranspiration data were combined to determine water sensitivity. Diurnal fluxes of gas exchange (net photosynthesis rate, transpiration rate, stomatal conductance, internal and atmospheric carbon ratio, water use efficiency and leaf temperature) and Chl a fluorescence (Fv/Fm, ΦPSII, ETR, Fv’/Fm’, NPQ and qL) were evaluated. Water restriction caused reductions of 90.9 and 84.7% in the increase in height and diameter of seedlings subjected to 25% water replacement when compared to seedlings with 100% water replacement, while biomass accumulation reduced by 96.9%. The kc values increased throughout the seedling production cycle, ranging from 0.59 to 2.86. Maximum water sensitivity occurred at 50% water replacement, with Ky = 1.62. Maximum carbon assimilation rates occurred in the morning, ranging from 6.11 to 12.50 µmol m-2 s-1. Ipê-rosa seedlings regulate the physiology of growth, gas exchange, and Chl a fluorescence depending on the amount of water available, and only 25% of the water re-placement in the substrate allows the seedlings to survive.

Harmful algal blooms are a serious problem for humanity, so the development of tools and methods for their timely detection is an urgent task. The use of optical methods often allows scientists to solve problems of remote and automated monitoring. Temperature spectra of fluorescence can be used as distinctive features of algae, and the fluorescence parameters obtained from them can be used for identification. The paper presents a description of the fluorescence parameters of Alexandrium catenella, and shows their stability during cumulative cultivation. In addition, the cytometric properties of the culture and its pigment composition were studied. In addition, the article compares all the studied properties of this microalga with the alga Prorocentrum cordatum. We demonstrated clear differences in their fluorescence parameters, which makes it possible to distinguish these microalgae by optical methods, for example, during their dominant blooms.

Background: Helicobacter pylori (H. pylori) is a prevalent bacterial pathogen implicated in various gastroduodenal diseases, including chronic gastritis, peptic ulcer disease, and gastric cancer. The COVID-19 pandemic has significantly influenced antibiotic prescription practices, potentially exacerbating antibiotic resistance in H. pylori. Objective: This study aims to investigate antibiotic prescription during COVID-19 pandemic in Croatia and its possible impact on H. pylori antibiotic resistance, with a focus on Croatia. Methods: A comprehensive literature search was conducted to identify relevant studies from March 2020 to June 2024. The search strategy included terms related to COVID-19, antibiotic prescription, and Croatia. Studies were selected based on predefined eligibility criteria, focusing on observational research addressing antibiotic use during the pandemic. Results: The COVID-19 pandemic leads to significant fluctuations in community antibiotic consumption within the European Union/European Economic Area. In Croatia, antibiotic use in 2022 surpassed 2019 levels, driven by increased consumption of macrolides and other antibiotics. This surge, influenced by early pandemic treatment claims and penicillin shortages, contributed to rising antibiotic resistance in H. pylori. Resistance rates to clarithromycin and levofloxacin were notably high, driven by mutations in the 23S rRNA, gyrA, and gyrB genes. Discussion: The increased antibiotic use during the COVID-19 pandemic has highly likely complicated H. pylori eradication efforts, highlighting the necessity of judicious antibiotic use and robust antimicrobial stewardship. The pandemic underscored the need for new therapeutic strategies, optimized eradication regimens, and advanced diagnostic methods to manage bacterial infections effectively. Conclusions: The COVID-19 pandemic has significantly impacted antibiotic use and resistance patterns, posing new challenges for H. pylori eradication. Addressing these challenges requires a multifaceted approach, including the development of new drugs and advanced diagnostics, coupled with sustained efforts in antimicrobial stewardship to combat emerging resistance threats.

Candida albicans is present in oral health and has been linked with dental caries. So far, the behavior of the yeast in the presence of Streptococcus sanguinis has not been tested. The aim was to determine the effect of excess sugar as a cariogenic environment on the biological and physico-chemical parameters of single-species (C. albicans) and dual-species biofilms of C. albicans and S. sanguinis. An in vitro caries model was used. The biofilms were exposed to 10% sucrose or 0.9% NaCl for 5 min 3x/day/5 days. Viable cells, hyphal quantification, SEM, biofilm pH, H2O2 quantification and surface microhardness of enamel slabs were determined. Microbial viability and the number of hyphae were higher for sucrose exposure, in the dual-species biofilms induced by excess sugar and coexistence between the two microorganisms. Sucrose-exposed biofilms showed a progressive pH drop over the time course. H2O2 production was about 6 times higher for the dual-species biofilms. Enamel demineralization was higher in the single-species, suggesting an inhibitory effect due to S. sanguinis. Excess sugar increases the pathogenic potential of C. albicans biofilm. In dual-species biofilms, S. sanguinis prevents pronounced drops in pH, decreasing demineralization and stimulates the development of hyphae in the yeast.

BReast CAncer gene 1 (BRCA1) and BReast CAncer gene 2 (BRCA2) encode for tumor suppressor proteins which are critical regulators of the homologous recombination (HR) pathway, the most precise and important DNA damage response mechanism. Dysfunctional HR proteins cannot repair double-stranded DNA breaks in mammalian cells, a situation called HR deficiency. Since their identification, pathogenic variants and other alterations of BRCA1 and BRCA2 genes have been associated with an increased risk of developing mainly breast and ovarian cancer. Interestingly, HR deficiency is also detected in tumors not carrying BRCA1/2 mutations, a condition termed “BRCAness”. One of the main mechanisms causing the BRCAness phenotype is the methylation of the BRCA1/2 promoters and this epigenetic modification is associated with carcinogenesis and poor prognosis mainly among patients with breast and ovarian cancer. BRCA1 promoter methylation has been suggested as an emerging biomarker of great predictive significance, especially concerning Poly (ADP-ribose) Polymerase inhibitors (PARP inhibitor-PARPi) responsiveness, along with or beyond BRCA1/2 mutations. However, as its clinical exploitation is still insufficient, the impact of BRCA1/2 promoter methylation status needs to be further evaluated. The current review aims to gather the latest findings about the mechanisms that underline BRCA1/2 function as well as the molecular characteristics of tumors associated with BRCA1/2 defects, by focusing on DNA methylation. Furthermore, we critically analyze their translational meaning and the validity of BRCA methylation biomarkers in predicting treatment response and we suggest a diagnostic pipeline that could be implemented in liquid biopsy to aid precision pharmacotherapy in BRCA-associated tumors.

Inula viscosa is a widely used plant in traditional Mediterranean and Middle Eastern medicine for various illnesses. I. viscosa has been shown to have anticancerous effects against various cancers, but its effects against lung cancer have been under limited investigation. At the same time, I. viscosa is rich in terpenoids whose anti-lung cancer effects have been poorly investigated. This study aimed to examine the potential anticancerous properties of methanolic and aqueous extracts of stems and leaves of I. viscosa and its terpenoid-rich fraction against human lung cancer A549 cells. Results showed a high radical scavenging capacity and polyphenol and flavonoid contents of I. viscosa methanolic rather than aqueous extracts. In addition, leaves methanolic extracts (IVLM) showed the highest reduction in viability of A549 cells among all the extracts. IVLM also reduced the viability of human ovarian SK-OV-3, breast MCF-7, liver HepG2, and colorectal HCT116 cancer cells. A terpenoid-rich I. viscosa fraction (IVL DCM), prepared by liquid-liquid separation of IVLM in dichloromethane (DCM), displayed a substantial reduction in the viability of A549 cells (IC₅₀= 27.8 ± 1.5 µg/mL at 48 h) and the panel of tested cancerous cell lines but was not cytotoxic to normal human embryonic fibroblasts (HDFn). The assessment of IVL DCM phytochemical constituents using GC-MS analysis revealed 21 metabolites, highlighting an enrichment in terpenoids, such as lupeol and its derivatives, caryophyllene oxide, betulin, and isopulegol, known to exhibit proapoptotic, and antimetastatic functions. IVL DCM also showed robust antioxidant capacity and decent polyphenol and flavonoid contents. Furthermore, Western blotting analysis indicated that IVL DCM reduced proliferation (reduction of proliferation marker Ki67 and induction of proliferation inhibitor proteins P21 and P27), contaminant with P38 MAP kinase activation, and induced the intrinsic apoptotic pathway (P53/BCL2/BAX/ Caspase3/PARP) in A549 cells. IVL DCM also reduced the migration of A549 cells, potentially by reducing FAK activation. Future identification of anticancer metabolites of IVL DCM, especially terpenoids, is recommended. These data place I. viscosa as a new resource of herbal anticancer agents.

A new quinazolinone alkaloid, named peniquinazolinone A (1), as well as eleven known com-pounds, 2-(2-hydroxy-3-phenylpropionamido)-N-methylbenzamide (2), viridicatin (3), viridicatol (4), (±)-cyclopeptin (5a/5b), dehydrocyclopeptin (6), cyclopenin (7), cyclopenol (8), methyl-indole-3-carboxylate (9), 2,5-dihydroxyphenyl acetate (10), methyl m-hydroxyphenylacetate (11), conidiogenone B (12), was isolated from the endophytic Penicillium sp. HJT-A-6. The chemical structures of all compounds were elucidated by comprehensive spectroscopic analysis, including 1D, 2D NMR, and HRESIMS. The absolute configuration at C-13 of peniquinazolinone A (1) was established by applying the modified Mosher’s method. Compounds 2, 3, and 7 exhibited an op-timal promoting effect on the seed germination of Rhodiola tibetica at a concentration of 0.01 mg/mL, while the optimal concentration for compounds 4 and 9 to promote Rhodiola tibetica seed germination was 0.001 mg/mL. Compound 12 showed optimal seed-germination-promoting activity at a concentration of 0.1 mg/mL. Compared with the positive drug 6-benzyladenine (6-BA), compounds 2, 3, 4, 7, 9, and 12 could extend the seed-germination period of Rhodiola tibetica up to the 11th day.

Investigation of oomycete diversity in rice paddies of Fars Province in Iran led to the identification of two new Pythium sensu lato (s.l.) species as Globisporangium izadpanahii sp. nov. and Pythium banihashemianum sp. nov. The identification was based on morphological and physiological features as well as the phylogenetic analysis of nuclear (ITS and βtub) and mitochondrial (cox1 and cox2) loci using Bayesian inference and Maximum Likelihood. A major characteristic of G. izadpanahii was the production of globose hyphal swellings; this species did not produce vesicles and zoospores. The present paper describes formally these two new species and defines their phylogenetic relationships with other congeneric species. According to multiple gene genealogy analysis, G. izadpanahii grouped with other species of Globisporangium in the former clade G of Pythium s.l. and was closely related to both G. nagaii and the recently described G. coniferarum. The second species, designated P. banihashemianum, grouped with other species of Pythium sensu stricto in the former clade B of Pythium s.l. and according to the phylogenetic analysis shared an ancestor with P. plurisporium. In pathogenicity tests on rice seedlings, P. banihashemianum isolates were highly pathogenic causing severe root and crown rot, while G. izadpanahii isolates were not pathogenic.

Pollutants accumulate in aquatic habitats as a result of mining activities. Lemna gibba and Lemna minor were used in this study to examine the accumulation capacities of Mo+, Pb+, and Cu++ in the acid fluids of the copper mining. Duckweed family includes water plants like L. minor and L. gibba. These are tiny, delicate, free-floating aquatic plants. Two reactors were assigned to L. gibba and L. minor, respectively. Throughout the course of eight days, these plants and reactor water were gathered every day. The acid mine water's pH, temperature, and electric conductivity were also tested every day. The L. gibba and L. minor were cleaned, dehydrated, and burned in a drying oven for a whole day at 300 oC. ICP-MS (Inductively Coupled Plasma Mass Spectroscopy) was used to determine the Mo+, Pb+, and Cu++ content of the plant and water samples. The concentrations of Mo, Pb, and Cu in the acid mine fluids of the copper mining were 30±4, 260±12, and 15535±322 μg L−1, respectively.in that order. When it came to extracting Mo, Pb, and Cu from the acidic fluids of the copper mining, L. gibba and L. minor performed well and efficiently compared to control samples; they did so 29 and 177 times for Mo, 30 and 109 times for Pb, and 495 and 1150 times for Cu. Considering all these findings, it has been shown that L. gibba and L. minor have the ability to efficiently remove Mo, Pb and Cu in acid mine fluids and can be good rehabilitation plants for polluted waters.

Primary congenital hypothyroidism is readily diagnosed based on elevated plasma TSH levels. In contrast, in the rare disorders of thyroid hormone resistance, TSH and often also thyroid hormone levels are within the normal range. Thyroid hormone resistance is caused by defects in hormone metabolism, transport, or receptor activation and has the same severe consequences for childhood development as congenital hypothyroidism. A total of 23,522 data points from a large cohort of children and young adults were used to generate normal values and sex-specific percentiles for the ratio of free T3 to free T4 (fT3/fT4 ratio). The aim was to determine whether cases with genetically confirmed thyroid hormone resistance (MCT8, THRα, and SECSIBP2 defects) had an abnormal fT3/fT4 ratio. Indeed, we were able to demonstrate a clear separation of patient values for the fT3/fT4 ratio from normal and pathological controls (e.g., children with severe cerebral palsy). We therefore recommend the use of the fT3/fT4 ratio as a readily available screening parameter in children with developmental delay for the identification of thyroid hormone resistance syndromes. The fT3/fT4 ratio can be plotted on centile charts using our free online tool at http://www.thyroid-hormone-ratio.org that accepts various SI and non-SI units.

Advances in biological sciences have been applied in medicine, agriculture and industry with great benefit. Some recent trends in society, governance structures and communications, however, appear to be hampering the use of new scientific findings for controlling infectious diseases. These limitations are illustrated with examples related to (i) the adaptation of fresh water mosquito vectors of major arboviral diseases to salinity in coastal areas with consequences for disease transmission, (ii) understanding the implications of reduced dengue transmission during the COVID-19 lockdown for dengue control, (iii) causes underlying the rapid spread of the malaria vector Anopheles stephensi in South Asia and Africa, (iv) the application of serodiagnostic techniques for Lyme disease and tick-borne relapsing fever caused by tick-borne bacteria of the genus Borrelia, and (v) COVID-19 prevention.

Abstract Octreotide is the first synthetic peptide hormone, consisted of eight amino acids, that mimics the activity of somatostatin, a natural hormone in the body. During the past decades, advanced instrumentation and crystallographic software established X-ray Powder Diffraction (XRPD) as a valuable tool for extracting structural information of biological macromolecules. The latter was demonstrated by the successful structural determination of octreotide at a remarkably high d-spacing resolution (1.87 Å) (PDB code: 6vc1). This study focuses on the response of octreotide to different humidity levels and temperatures, with a particular focus on the stability of the polycrystalline sample. XRPD measurements were accomplished employing an Anton Paar MHC-trans humidity-temperature chamber installed within a laboratory X’Pert Pro diffractometer (Malvern Panalytical). The chamber is employed to control and maintain precise humidity and temperature levels of samples during XRPD data collection. Pawley analysis of the collected data sets revealed that the octreotide polycrystalline sample is remarkably stable and no structural transitions were observed. The compound retains its orthorhombic symmetry (space group: P212121, a = 18.57744(4) Å, b = 30.17338(6) Å, c = 39.70590(9) Å, d~ 2.35 Å). However, a characteristic structural evolution in terms of lattice parameters and volume of the unit-cell is reported mainly upon controlled relative humidity variation. In addition, an improvement of signal-to-noise ratio in the XRPD data under a cycle of dehydration/rehydration is reported. These results underline the importance of considering the impact of environmental factors, as humidity and temperature, in the context of structure-based drug design, thereby contributing to the development of more effective and stable pharmaceutical products.

Tick-borne encephalitis virus (TBEV) is a RNA-containing enveloped virus, a member of the Flaviviridae family. Here we describe a detailed analysis of the size and structure of inactivated TBEV (the Sofyin-Chumakov TBEV strain, used in vaccines). Four analytical methods were used to analyze individual TBEV particles—negative staining TEM, cryo-EM, AFM, and NTA. All methods confirmed that the particles were monodisperse, and their mean size was ~50 nm. Cryo-EM data were used to obtain a 3D electron density model of the virus with clearly distinguishable E-proteins. STEM-EELS analysis detected phosphorous in the particles, which was interpreted as the RNA signal. Altogether, the described analytical procedures can be valuable for the further analysis of vaccine inactivated virus samples.

Since the mid-1990s probiotics have been proven to support the treatment of several gastrointestinal diseases. Additionally, probiotics have been taken as a common panacea for civilization diseases. To date, the only probiotic yeasts with evidence of health-promoting effects are Saccharomyces cerevisiae var. boulardii. The probiotic properties are strain-dependent, whereby it is crucial to develop methods to verify the probiotic-enriched food authentication including the growing number of dietary supplements with simplified commercialisation procedures. Due to the significant genetic similarity of S. cerevisiae var. boulardii to S. cerevisiae, qPCR-HRM analysis was tested, which has a very high sensitivity for polymorphism detection and enables simultaneous identification of the microorganism in the presence of the reference sample. The effectiveness of interspecies primer pairs and intragenus primer pairs designed to amplify heterogeneous regions was examined. HRM analysis using primers for 18SrRNA and ITS allowed for identification of S. cerevisiae var. boulardii at the species level. S. cerevisiae var. boulardii was efficiently identified at the variety level with amplification of HO and RPB2 in the single- and multi-component DNA extracts. The RPB2 amplicon showed the highest intraspecies differentiation power. The predominance of S. cerevisiae var. boulardii in the mixtures tested was crucial for identification using qPCR-HRM analysis and quantitatively corresponded to the strain content in the probiotic-enriched food matrices.

Excess nitrogen in agricultural drainage poses a serious threat to the water quality safety of the Yellow River basin. Utilizing aquatic plants to modify the rhizosphere microbial community structure and facilitate nitrogen transformation is a crucial strategy for mitigating regional water eutrophication. Our results revealed that the denitrification rate (DR), dissimilatory nitrate reduction to ammonium rate (DNRAR) and nitrogen fixation rate (NFR) of rhizosphere sediments exhibited a significant increase in artificially cultivated ditch. Compared with those in natural ditches, the richness of the bacterial community and the relative abundances of Bacteroidota, Verrucomicrobiota, Firmicutes, Anaeromyxobacter and Geobacter, which are involved mainly in the nitrogen cycle, in the rhizosphere sediments of artificially cultivated ditch were significantly greater due to improvements in environmental conditions. In artificially cultivated ditch, the dominant functional genes affecting DRNARs in the rhizosphere sediments of P. australis were nrfC and nrfA, while DRs were driven mainly by norB and napA, which were influenced by the nitrogen and carbon levels; and the dominant functional genes affecting NFRs in the rhizosphere sediments of T. orientalis were nifD, nifK and nifH. Our results provide a scientific basis for the use of aquatic plants for mitigating excess nitrogen levels in agricultural drainage.

Strong-aroma Baijiu is a traditional Chinese spirit distilled from grains that undergo solid-state fermentation. At the end of each fermentation cycle, a significant quantity of Huangshui accumulates at the bottom of the fermentation pit, constituting one of the primary by-products in the brewing process of strong-flavor liquor. In this report, the chemical component analysis of Huangshui obtained from a strong-flavor liquor factory revealed a diverse array of organic acids, alcohols, esters, starch, reducing sugars, amino acids, and other compounds present in Huangshui. Additionally, two specialized solid-state fermentation models were developed to investigate the formation process of Huangshui. Our findings demonstrate a positive correlation between the yield of Huangshui and the initial moisture content of fermented grains. It is concluded that Huangshui is the percolate produced during the solid-state brewing process of strong-flavor liquor, with its water content originating from the initial moisture of fermented grains.

Bats are natural hosts for a wide variety of viruses and could therefore be the reservoir, source and spillover for several mammalian viruses, including adenoviruses. We examined faecal samples from 12 bats (five species of the family Vespertilionidae) from central Russia (Moscow Region) and found that two of them (16%) were positive for mastadenoviruses. One of the identified viruses was isolated from faeces of Nyctalis noctule and named BatAdV/MOW15-Nn19. The genome of the BatAdV/MOW15-Nn19 was 37,915 bp in length. Comparative studies have shown significant divergence of the nucleotide sequence of the BatAdV/MOW15-Nn19 genome from European mastadenoviruses, the highest genome sequence similarity was to isolate WA3301 from an Australian bat. Phylogenetic analysis, as well as a comparative analysis of the protein coding genes to homologous available from public database, showed evidence of novel species within Mastadenovirus genus (preliminary named «K»). Analysis of the short part of the DNA polymerase gene of BatAdV/MOW15-Nn19 reveals few NCBI records from Europe with high identity to the new virus, i.e. mastadenoviruses related to the new species are circulating in European bats. The partial genome of another mastadenovirus from Pipistrellus nathusii was identified, it represented bat adenovirus species B, which is widely distributed in European Vespertilionidae bats. New data on the genetic diversity of bat adenoviruses may help to better understand how they evolved in mammals and spread across Europe.

(S)-Atenolol ((S)-2-(4-(2-Hydroxy-3-(isopropylamino)propoxy)phenyl)acetamide) has been synthesized in >99% ee with use of Candida antarctica lipase B from Syncozymes (Shanghai, China), in a kinetic resolution of the corresponding racemic chlorohydrin. Deprotonation of the starting phenol building block was base catalyzed. The enantiopurity of the chlorohydrin building block remained unchanged upon subsequent amination. All four steps in the synthesis protocol have been optimized compared to previously reported methods, which makes this new protocol more sustainable and in accordance with green chemistry principles compared to previously reported protocols.

An extensive thermodynamic study of N-methylformamide (CAS RN: 123-39-7) and N,N-dimethylformamide (CAS RN: 68-12-2), is presented in this work. The liquid heat capacities of N-methylformamide were measured by Tian–Calvet calorimetry in the temperature interval (250 – 300) K. The vapor pressures for N-methylformamide and N,N-dimethylformamide were measured using static method in the temperature range 238 K to 308 K. The ideal-gas thermo-dynamic properties were calculated using a combination of the density functional theory (DFT) and statistical thermodynamics. A consistent thermodynamic description was developed using the method of simultaneous correlation, where the experimental and selected titerature data for vapor pressures, vaporization enthalpies, and liquid phase heat capacities and calculated ide-al-gas heat capacities are treated together to ensure overall thermodynamic consistency of the results. Resulting vapor pressure equation is valid from the triple point to the normal boiling point temperature.

This study aimed to determine gene pool structure in the tomato (Solanum lycopersicum) along with the relationships of populations in different regions. To do so, it leveraged 276 diverse cultivated accessions representing 35 countries, obtained via the USDA-GRIN tomato germplasm collection. Single nucleotide polymorphisms (SNPs) in these accessions were detected by sequencing, and genetic diversity, population structure, and phylogenetic relationships were estimated from the SNP alleles. The results of model-based ancestry analysis, phylogenetic tree construction, and principal component analysis supported these accessions as representing three well-differentiated genetic populations. Phylogenetic analyses traced the accessions back to their regions of origin, highlighting Central and South America as the primary center of domestication, with secondary centers in Europe and North America. This study provides a comprehensive analysis of the genetic variation in and relationships among cultivated tomato genotypes. The findings are of benefit to curators, researchers, and breeders in understanding, utilizing, conserving, and managing the collection, and in thereby contributing more efficiently to international tomato research.

Tomato (Solanum lycopersicum L.) is one of the most commercially essential vegetable crops cultivated worldwide. In addition to the nutritional value, tomato is an excellent model for studying climacteric fruits' ripening process. Despite this, the available natural pool of genes that allows expanding phenotypic diversity is limited, and the difficulties of crossing using classical selection methods when stacking traits increase proportionally with each additional feature. Genetic engineering makes it possible to introduce changes in genes of interest without changing the allelic combinations characteristic of successful commercial varieties, to enhance the expression of existing genes, to create and transfer artificial and heterologous genes, and much more. However, this requires understanding the fundamental principles of the molecular interaction. Although the leading candidate genes for these components have been identified, a complete picture of their relationship has yet to be formed. The review summarizes the latest (2017-2023) achievements related to studying the ripening processes of tomato fruits. In this work, an attempt is made to systematize the results of various research and display the interaction pattern of genes regulating the process of tomato fruit ripening.

Microbial-based biostimulants containing arbuscular mycorrhizal fungi (AMF), Trichoderma fungi and plant growth-promoting rhizobacteria (PGPR) have been applied in an open-field tomato cultivation. A two-years field experiment (2020-2021) was performed in Southern Italy on “Heinz 1534” processing tomato hybrid, using three commercial formulations characterized by different microbial consortia (MIC: Glomus spp., Rhizophagus spp., Bacillus spp., Streptomyces spp., Pichia spp., Trichoderma spp.; EKO: Glomus spp., Bacillus spp., Streptomyces spp., Pseudomonas spp., Arthrobotrys spp., Monacrosporium spp., Paecilomyces spp., Myrothecium spp., Trichoderma spp.; FID: Glomus spp., Bacillus spp., Trichoderma spp.) and comparing them to untreated control (CTRL). The effect of growing season and microorganism-based treatments on yield, technological traits and functional quality of tomato fruits was assessed. The year of cultivation (Y) affected yield (with lower fruit weight, higher marketable to total yield ratio and higher % of total defected fruits in 2020), and technological components (higher dry matter, total acidity, total soluble solids in 2020). During the first year of the trial, all evaluated treatments (MIC, EKO and FID) enhanced soluble solids content by 10%, on average, compared to CTRL. Sucrose and lycopene contents were influenced both by the microbial-based treatments and the growing season (greater values found in 2021 with respect to the first year). Y factor also significantly affected all evaluated metabolites contents, except for tyrosine, essential (EAA) and branched-chain (BCAAs) amino acids. Over two years of field trial, FID biostimulant enhanced the contents of proteins (+53.71%), alanine (+16.55%), aspartic acid (+31.13%), γ-aminobutyric acid (+76.51%), glutamine (+55.17%), glycine (+28.13%), monoethano-lamine (+19.57%), total amino acids (+33.55), essential amino acids (+32.56%) and branched-chain amino acids (+45.10%) compared to the untreated control. Our findings highlighted the valuable effect of FID microbial inoculant in boosting several primary metabolites (proteins and amino acids) in the fruits of processing tomato crop grown under Southern Italy environmental condi-tions, although no effect on yield and its components was appreciated.

Argulus is an ectoparasite that frequently infects goldfish, with severe effects and a high death rate. The use of plant extracts has been widely reported to have antiparasitic activities, including one of the solasodine compounds in sour eggplant extracts. This research evaluated the antiparasitic activities of Compositions A (Solanum ferox and Zingiber zerumbet extract at 400 and 200 ppm, respectively), B (SFE and Boesenbergia pandurata at 900 ppm), and C (single Solanum ferox at 400 ppm). The research stages comprised 1) isolation and analysis of solasodine content in sour eggplant extracts and its extract composition, 2) in vitro antiparasitic activities on Argulus sp. at doses of 50 and 100 ppm with a testing time of 60–240 minutes, and 3) in vivo antiparasitic activities on Argulus sp. on goldfish at a dose of 100 ppm and observation for 2–12 hours of medication. The results showed that sour eggplant extracts had a solasodine concentration of 7.151 mg/L, whereas the derivative compositions A, B, and C were 656, 485, and 295 ppm, respectively. The in vitro testing demonstrated that composition A was effective for killing approximately 80–100% of Argulus, whereas compositions B and C killed 80–90% and 60–70%, respectively. Viewed from the effectiveness for killing parasites, the result was excellent (above 50%). The in vivo medication test was continued using three extract compositions at a dose of 100 ppm. Compositions A, B, and C were found to be capable of releasing 81.33, 75.67, and 71.00 arguli, respectively, per fish. We concluded that the single SFE extract and a concoction with BPE and ZZE had reasonable antiparasitic activity, whereas the concoction of SFE and ZZE killed more Argulus parasites at a higher rate.

The goal of germplasm enhancement is to introgress traits from wild crop relatives into cultivated material and eventually cultivars. It seeks to restore genetic diversity that has been lost over time or to augment cultivated material with novel alleles that improve parents in breeding programs. This paper discusses potato germplasm enhancement efforts in the past, focusing on effective examples such as disease resistance and processing quality. In addition, it outlines new strategies for enhancement efforts, shifting the focus from evaluating phenotypes to tracking and manipulating specific DNA sequences. In the genomics era, germplasm enhancement will increasingly be focused on identifying and introgressing alleles rather than traits. Alleles will come from a broad pool of genetic resources that includes wild species relatives of potato, landraces, cultivated potato itself, and distantly related species. Genomics tools will greatly increase the efficiency of introgressing multigenic traits, and will make it possible to identify rare alleles and utilize recessive alleles.

Plants grown under fluctuating light impact plant developments compared with those grown under non-fluctuating light conditions. However, our knowledge on the underlying regulatory mechanisms is still quite limited, particularly from the transcriptional perspective. In order to investigate the influence of different light intensity distributions on tomato plant development, we designed three fluctuating light intensity distributions with the non-fluctuating light intensity as control and compared the transcriptional differences after five weeks of treatment. We found plant height and aerial/root weight were significantly reduced under all fluctuating light treatments. Transcriptome analysis revealed that the number of up and down regulated genes had a distinct distribution pattern between different treatments and control. The largest difference between the numbers of down and up regulated genes was found between treatment 1 and 3, reaching to a total of 416 genes. The number and type of the top 20 enriched pathways differed between treatments and control. The largest number of genes enriched was involved in the biosynthesis of secondary metabolites. These results provide insights into the transcriptional regulations of tomato under different light intensity distributions.

Variations in liver metabolism as a result of hepatitis C virus have been established by numerous clinical trials. The use of antioxidants supplements has been reported to minimize the implication of this disease. In this regard, we examined the suitability of Solanum fruit juice, a natural source of vitamin C and citrus flavoniod as a precursor for the treatment of patients with chronic hepatitis C. Forty adult patients who were diagnosed with chronic hepatitis C and were under antiviral therapy were divided into two equal groups. Group 1 patients received their antiviral therapy with normal food and water and served as the control group while patients in group 2 were supplemented with Solanum fruit juice for eight consecutive weeks. Measurements for Anthropometric data, C reactive protein (CRP), atherogenic indices, biochemical parameters and activities of liver marker enzymes were recorded before and after eight weeks. No alterations were found in waist circumference, body mass and body fat following regular use of Solanum fruit juice. The serum levels of oxidative stress markers, LDL-cholesterol, total cholesterol, CRP and atherogenic indices decreased in the Solanum fruit juice group when compared to the control group. Moreover, the activities of the liver marker enzyme AST decreased in those who had high levels before the intervention. These results underscore the benefits of Solanum fruit juice in the diet of patients with HCV as a result of decreased cholesterol in blood serum, decreased inflammation, and increase in antioxidant capacity as well as maintaining body mass index. This clinical trial is registered at Pan African Clinical Trial Registry (www.pactr.org) with unique identification number PACTR201802003092138.

Anxiety is a multifactorial pathology associated with oxidative stress and changes in different neurotransmitters in the central nervous system (CNS). Sarcomphalus joazeiro (Rhamnaceae) has a predicted effect on the CNS, being considered an alternative for pre-clinical investigations of anxiolytic drugs. The objective of this study is to evaluate the chemical composition, analyze the antioxidant capacity and the anxiolytic effect of the ethanolic extract of S. joazeiro leaves (EEFSJ) in zebrafish. The chemical profile was analyzed by liquid chromatography coupled to mass spectrometry (UPLC-PDA-ESI-QDA). Antioxidant action by DPPH• elimination and ABTS•⁺ capture assays. The 96 h acute toxicity and open field and light and dark tests, for sedative and anxiolytic evaluation, respectively, were applied in vivo, using zebrafish. It was possible to identify 11 compounds from the flavonoid and saponin class. EEFSJ exhibited a mean inhibitory concentration (IC50) of 185.2 ± 2.2 µg/mL and 74.17 ± 1.5 µg/mL against the DPPH• and ABTS•⁺ radicals, respectively. In vivo, EEFSJ demonstrated reduced locomotor activity and an anxiolytic effect similar to diazepam via GABAergic systems, without presenting toxicity within 96 h. These results suggest the safety of using EEFSJ for the development of clinical trials for the production of new anxiolytic drugs.

This study was performed to evaluate the efficacy of nicotinic acid against Streptococcus pneumoniae biofilm. The experimental results indicate that S. pneumoniae forms biofilms, particularly at the liquid-air interface, as evidenced by the pellicle assay. Nicotinic acid demonstrated significant inhibition of S. pneumoniae biofilm formation at 6 h and 18 h, with higher concentrations leading to lower biofilm biomass, suggesting a concentration-dependent response. However, minimal biofilm inhibition was observed at 12 h. Treatment with nicotinic acid induced FTIR spectral changes in protein biomolecules of S. pneumoniae biofilm at 6 h and 18 h, evidenced by alterations in spectral peaks at 750 cm-1, 906 cm-1, 1540 cm-1, 1548 cm-1, and 1644 cm-1. Notably, no FTIR spectral changes were detected in 12 h S. pneumoniae biofilm following exposure to nicotinic acid. These findings suggest that nicotinic acid inhibits biofilm formation in a time and concentration-dependent manner, possibly by altering the structure of protein biomolecules within the biofilm.

The occurrence and ability of Salmonella typhimurium to withstand the effects of various antibiotics has been extensively recorded. This highlights the necessity for alternative methods to combat Salmonella infections. The current investigation aimed to ascertain the impact of benzoic acid on the extent of surface coverage and biomass of S. typhimurium biofilm after 48 and 72 hours. The findings indicated that all the different doses of benzoic acid (ranging from 3.125% to 50%) resulted in a decrease in the extent of biofilm surface coverage. Benzoic acid exhibited a substantial inhibitory effect on the biofilm biomass at all concentrations. Higher concentrations of benzoic acid led to a decrease in biofilm biomass, suggesting a concentration-dependent response. This finding implies that benzoic acid is effective in managing Salmonella infections.

Gut colonization by multidrug resistant organisms(MDROs) is a risk factor for bloodstream infections(BSI) in the early phase of Hematopoietic Stem Cell Transplantation(HSCT). We evaluated, in an exploratory study, the impact of Lactobacillus plantarum on modulation of the gut microbiome in HSCT patients colonized by MDROs. Participants were allocated to an intervention group(IG=22) who received capsules of L. plantarum(5x109 CFU) twice a day until the onset of neutropenia and a control group(CG=20). The V4 region of the 16S bacterial rRNA gene in 93 stool samples from a subset of 33 patients was sequenced(IG=20 and CG=13). Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) (PICRUSt2) program was used for prediction of metagenome functions. L. plantarum had an average 86%(±11%) drug-target engagement at 43(±29) days of consumption and it was safe, tolerable and associated with an increase in the abundance of the Lactobacillales(p=0.004). A significant increase of Lactococcus and reduction of Turicibacter(p<0.05) were identified on the second-week of L. plantarum use. Although Enterococcus abundance had a greater rise in the CG (p=0,07), there were no significant differences concerning the gram-negative MDROs. No serious adverse effects was reported in the IG. . We observed greater non-significant pyruvate fermentation to propanoate I (p=0.193) relative abundance in the IG comparing with CG. L. plantarum use was safe and tolerable by HSCT patients. Although it may have impact on Enterococcus abundance and reduction of Turicibacter, it has not showed impact on Gram-Negative MDRO abundance in MDRO gram-negative colonized HSCT patients.

Structurally clear compounds have advantages for quality control in plant biostimulant production and application processes. Humic acid (HA) is a biostimulant that significantly affects plant growth, and soil-dwelling Protaetia brevitarsis larva (PBL) can rapidly convert agricultural waste into HA. In this study, we use PBL as a model to investigate HA formation and screen for HA-related structurally clear plant biostimulant compounds. The dephasing magic angle spinning nuclear magnetic resonance (13C DD–MAS NMR) analysis indicated HA structural changes during PBL digestion; the metabolic profile detected seven HA-related aromatic ring-containing compounds. Plant experiments identified six compounds with stimulating effects, with piperic acid being reported for the first time in this study. Thus, this study not only provides insights into the insect-mediated transformation of HA but also illustrates a new strategy for discovering structurally clear plant biostimulant compounds.

Candida glabrata (Nakaseomyces glabratus), the second most prevalent Candida pathogen globally, has emerged as a major clinical threat due to its ability to develop high-level azole resistance. In this study, two new 5,6-dihydrotetrazolo[1,5-c]quinazoline derivatives (c11 and c12) were synthesized and characterized using IR, LC-MS, 1Н and 13C NMR spectra. Along with 13 previously reported analogues, these compounds underwent in vitro antifungal testing against clinical C. glabrata isolates using a serial dilution method (0.125-64 mg/L). Remarkably, compounds c5 and c1 exhibited potent antifungal activity, with minimum inhibitory concentrations of 0.37 μM and 0.47 μM, respectively – about 20-fold improvement over standard drugs like amphotericin B, caspofungin, and micafungin. A detailed structure-activity relationship analysis revealed crucial molecular features enhancing antifungal potency. Extensive molecular docking studies across 18 protein targets explored potential binding pockets and affinities of the lead compounds. A robust 3D-QSAR model, incorporating molecular descriptors Mor26m and Mor29e, displayed good predictive ability for antifungal activity. In silico predictions indicated an absence of herbicidal effect, negligible environmental toxicity (to honeybees, avian species, and aquatic organisms), and mild human toxicity concerns for these compounds. This comprehensive approach aims to develop novel and effective antifungal compounds against the clinically relevant pathogen C. glabrata.

Helicobacter pylori infection constitutes a silent pandemic of global concern. In the last decades, the alarming increase of multidrug resistance evolved by this pathogen has led to a marked drop in the eradication rates of traditional therapies worldwide, encouraging scientific community to discover and develop new antimicrobial strategies. In the present work, we identified a battery of novel drug-like HsrA inhibitors with MIC values ranging from 0.031 to 4 mg/L against antibiotic-resistant strains of H. pylori. Most of these novel antimicrobials exhibited minor effects against both Gram-negative and Gram-positive species of human microbiota. The most potent anti-H. pylori candidate demonstrated a high therapeutic index, additive effect in combination with metronidazole and clarithromycin, as well as a strong antimicrobial action against Campylobacter jejuni, another clinically relevant pathogen of phylum Campylobacterota. Transcriptomic analysis suggested that in vivo inhibition of HsrA triggered lethal global disturbances in H. pylori physiology, including the arrest of protein biosynthesis, malfunction of respiratory chain, detriment in ATP generation, and oxidative stress. The novel drug-like HsrA inhibitors described here constitute valuable candidates to a new family of narrow-spectrum antibiotics that allow overcoming the current resistome, protecting from dysbiosis, and increasing therapeutic options for novel personalized treatments against H. pylori.

Rare Earth Elements (REE), also known as Lanthanides (Ln3+), are a group of 17 elements showing peculiar physical and chemical properties. Unlike technological applications, very little is known about the physiological role and toxicity of Ln3+ on biological systems, in particular on microorganisms (e.g., bacteria), which represent the most abundant domains on our planet. Up to now, very limited studies have been conducted due to Ln3+ precipitation with some anions commonly present in the culture media. Therefore, the development of a minimal medium is essential to allow the study of Ln3+-microbial interactions, limiting considerably the precipitation as insoluble salts. In this regard, a new minimal culture medium capable of solubilizing large amounts of Ln3+, and allowing the growth of different microbial taxa, was successfully developed. Assays have shown that the medium is capable of solubilizing Ln3+ up to 100 times more than other common culture media and allowed the growth of 63 bacteria and 5 fungi. The kinetic growth of one yeast and one gram-positive bacterium has been defined, proving to support superior growth and biomass compared to other commonly used minimal media. Moreover, the sensitivity and uptake/absorption of a Bacillus stratosphericus strain was tested, highlighting its capability to tolerate concentrations up to 10 mM of either Cerium, Gadolinium or Lanthanum and accumulate different quantities of the three.

Chaga (Inonotus obliquus) is a pathogenic fungus that grows mostly on birch species (Betula pendula and B. pubescens) and has traditionally been used as an anti-cancer medicine. The study aimed to compare the chemical composition and cytotoxic activity of chagas growing on both Betula spp. on various cancer cell lines. The freeze dried extracts contained triterpenes inotodiol, lanosterol betulin and betulinic acid typical to conks growing on Betula species. The cytotoxic activity of chaga growing on Betula pendula and B. pubescens 80% ethanolic extracts against thirty-one human cancer cell lines has been evaluated by a sulforhodamine B assay. The strongest inhibitions were observed with chaga (growing on B. pendula) extract on the HepG2 and CAL-62 cell line, and with chaga (from B. pubescens) extract on HepG2 cell line, with IC50 values of 37.71, 43.30, and 49.99 μg/mL, respectively. The chaga extracts from B. pendula exert somewhat stronger effects on most cancer cell lines studied than B. pubescens extracts. This study highlight the potential of chaga as a source of bioactive compounds with selective anti-cancer properties, encouraging further exploration of its therapeutic mechanisms and potential applications in cancer treatment.

Cucurbitacin phytonematicides consistently suppress population densities of plant nematodes, but have no effect against bacteria, fungi and insects. This study investigates the mechanism through which cucurbitacin phytonematicides consistently exert nematicidal properties on nematodes. Post 72-h exposure of second-stage juveniles (J2) of Meloidogyne enterolobii to geometric concentration of Nemafric-BL phytonematicide, J2 total proteins were measured using TruspecCHNS Macro. Total proteins versus phytonematicide exhibited a negative quadratic relation (R² = 0.97), with the minimum total proteins achieved at x = 4.9% phytonematicide. Beyond the minimum, total proteins increased gradually, depicting the existence of both isoprenylation (breakdown) and farnesylation (biosynthesis) of total proteins. In conclusion, cucurbitacins and nematode cuticles are both chemically structured, which provides the mechanism through which the test chemicals consistently decrease population densities of plant nematodes.

Banana is one of the most important fruits in the world due to its status as a major food source for more than 400 million people. Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) causes substantial losses of banana crops every year, and molecular host resistance mechanisms are currently unknown. We here performed a genome-wide analysis of the autophagy related protein 8 (ATG8) family in a wild banana species. The banana genome was found to contain 10 MaATG8 genes. Four MaATG8s formed a gene cluster in the distal part of chromosome 4. Phylogenetic analysis of ATG8 families in banana, Arabidopsis thaliana, Citrus, rice, and ginger revealed five major phylogenetic clades shared by all of these plant species, demonstrating evolutionary conservation of the MaATG8 families. The transcriptomic analysis of plants infected with Foc TR4 showed that almost all of the MaATG8 genes were more highly induced in resistant cultivars than in susceptible cultivars. Finally, MaATG8F was found to interact with MaATG4B in vitro (with yeast two-hybrid assays), and MaATG8F and MaATG4B all positively regulated banana resistance to Foc TR4. Our study provides novel insights into the structure, distribution, evolution, and expression of the MaATG8 family in bananas. Furthermore, the discovery of interactions between MaATG8F and MaATG4 could facilitate future researches of disease resistance genes for genetic improvement of bananas.

Background: Patients with mutations in the monocarboxylate transporter 8 (MCT8) suffer from Allan-Herndon-Dudley syndrome (AHDS), characterized by developmental delay and a highly disabling movement disorder. Despite the potential of thyroid hormone derivatives to overcome the transporter defect, current trials did not achieve patient-oriented therapeutic goals. Objectives: Since most neurological symptoms are related to the dopaminergic system, we investigated the role of dopamine and its metabolites in MCT8 deficiency with regard to pathophysiology and potential therapeutic strategies in an observational cohort study. Methods: We present longitudinal data from the DEEPTYPE registry of ten patients with video-documentation, standardized phenotyping, cerebrospinal fluid analysis, treatment response to levodopa/carbidopa supplementation, and neuroimaging data. To establish a cell-based model for pathophysiological studies, we differentiated healthy human induced pluripotent stem cells (hiPSCs) into dopaminergic neurons. Results: Children presented with signs of parkinsonism in childhood, including hypokinesia, hypomimia, inability to sit or stand, rigidity, dystonia, and autonomic dysfunction along the classification of Leuzzi and colleagues. Cerebrospinal fluid homovanillic acid concentrations were decreased (n=11), suggesting isolated dopamine pathway impairment. Six out of seven patients responded favorably to levodopa/carbidopa supplementation and we did not see any adverse drug reactions. Our cell-based studies showed that hiPSC-derived dopaminergic neurons expressed MCT8 and produced quantifiable levels of biogenic amines. Conclusions: Parkinsonism is part of AHDS's clinical presentation and may be amenable to treatment. The precise impact of MCT8 deficiency on the dopamine metabolism needs to be further elucidated, e.g. by using patient iPSC-derived dopaminergic neurons in future studies.

Our environment has been impacted by man-made pollutants mainly industries make substantial use of synthetic dyes which exhibit cytotoxicity and have significant environmental consequences. Effective photocatalyst-based approaches for degrading synthetic dyes into less toxic chemical are of great interest. Synthesizing NPs using biological approaches, particularly plant-based approaches offer advantages, decreasing the risk of NPs losing biocompatibility during synthesis, cost-effectiveness, and eco-friendliness. In this study, we employed a green synthesis method to produce manganese oxide nanoparticles (MnO NPs) utilizing leaf extract from the Lathyrus Aphaca plant. The synthesized MnOx NPs were characterized through various techniques; X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), Fourier-transform infrared spectroscopy (FTIR), and UV–visible spectroscopy. XRD analysis showed distinct peaks, indicated the presence crystallographic planes within the MnO₂ nanoparticles, thus confirming their crystalline structure. SEM study highlighted closely packed configurations with average size of approximately ≥ 50-70 nm, with well-defined boundaries. EDX analysis showed the presence of elemental manganese (Mn) in the samples, with peaks corresponding to Mn detected at lower energy levels, predominantly represented by MnO. FTIR, showed the presence of the O-O stretching mode at a frequency of 719 cm^-1, the presence of MnO₆ oxides of manganese, and peak at 548 cm^-1 corresponded to the Mn-O stretching mode. Furthermore, the green-synthesized manganese oxide nanoparticles exhibited promising photocatalytic and adsorption capabilities against Methylene Blue (MB) dye, leading to approximately 93.21% degradation of MB when treated with the green-synthesized MnO nanoparticles derived from plant extract. This highlights the efficacy and potential of these nanoparticles in environmental remediation applications, particularly in the degradation of methylene blue contaminants.

Abstract: Pasteurella spp. is a gram-negative bacterium that is part of the oral and upper respira-tory tract microbiota of many animals such as canids and felines. In humans it can produce pa-thology primarily associated with animal bites or scratches. In addition, members of this genus, and especially Pasteurella multocida, may also be involved in systemic infections, mainly in im-munocompromised patients in close contact with pets. The aim of this study is to analyse the prevalence and clinical characteristics of infections caused by Pasteurella spp. as well as the sensitivity profile of these isolates obtained in the Microbiology laboratory of the Hospital Can Misses (Ibiza, Ibiza and Formentera Health Area, ASEF) in the period from January 2013 to December 2018. The group of islands of Ibiza and Formentera is known as the Pitiusas Islands. Retrospective descriptive study in which all Pasteurella spp. isolates were analysed in the Microbiology Laboratory of Hospital Can Misses from 1 January 2013 to 31 De-cember 2018. Bacterial identification was performed by Vitek 2 automated biochemical test panel (bioMérieux, Spain) and antimicrobial susceptibility by E-test or antibiotic discs in Mueller-Hinton blood agar, interpreted based on European Committee on An-timicrobial Susceptibility Testing (EUCAST) standards, 2022 version. Medical records were systematically reviewed by collecting demographic data of infected patients, comorbidities, epidemiological data and clinical features of the infection. A total of 22 isolates of Pasterurella spp. were obtained from 22 different patients, 62.8% female, from three different species: 18 P. multocida, 2 P. canis and 2 P. pneumo-tropica. Most isolates came from soft tissue infection samples: 7 wound exudates and 5 abscess material. The antibiotics tested with the highest sensitivity profile were gen-tamicin and cefepime (100% sensitivity). The species most commonly isolated in our environment is P. multocida. Close contact with companion animals, especially cats and dogs, is the main risk factor for Pasteurella spp. infections, although in many cases such contact cannot be established. So Pasteurella spp. infection should be considered a zoonosis in possible emergency, especially in rural areas such as Pitiusas Islands.

Antimalarial drug resistance has become a real public health problem despite WHO measures. New sequencing technologies make it possible to investigate genomic variations associated with resistant phenotypes at the genome-wide scale. Based on the use of hemisynthetic nanopores, the PromethION technology from Oxford Nanopore Technologies can produce long-read sequences, in contrast to previous short-read technologies used as gold-standard to sequence Plasmodium. Two clones of P.falciparum (Pf3D7 and PfW2) were sequenced in long-read using the PromethION sequencer from Oxford Nanopore Technologies without genomic amplification. This made it possible to create a processing analysis pipeline for human Plasmodium with ONT Fastq only. De novo assembly revealed N50 lengths of 18,488 kb and 17,502 kb for the Pf3D7 and PfW2, respectively. The genome size was estimated at 23,235,407 base pairs for the Pf3D7 clone and 21,712,038 base pairs for the PfW2 clone. The average genome coverage depth was estimated at 787X and 653X for the Pf3D7 and PfW2 clones, respectively. This study proposes an assembly processing pipeline for the human Plasmodium genome using software’s adapted to large ONT data and the high AT percentage of Plasmodium. This search provides all the parameters, which have been optimized, for use with the software selected in the pipeline.

In an era of increasing interest in the potential health benefits of medicinal foods, the need to assess their safety and potential toxicity remains a critical concern. While these natural remedies have garnered substantial attention for their therapeutic potential, a comprehensive understanding of their effects on living organisms is essential. We examined 316 herbal extracts to determine their potential nematocidal attributes in Caenorhabditis elegans. Approximately 16% of these extracts exhibited the capacity to induce diminished survival rates and larval arrest, establishing a correlation between larval arrest and overall worm viability. Certain extracts led to an unexpected increase in male nematodes, accompanied by a discernible reduction in DAPI-stained bivalent structures and perturbed meiotic advancement, thereby disrupting the conventional developmental processes. Notably, Onobrychis cornuta and Veratrum lobelianum extracts activated a DNA damage checkpoint response via the ATM/ATR and CHK-1 pathways; hindering germline development. Our LC‒MS analysis revealed jervine in V. lobelianum and nine antitumor compounds in O. cornuta. Interestingly, linoleic acid replicated phenotypes induced by O. cornuta exposure, including an increased level of pCHK-1 foci, apoptosis, and the MAPK pathway. We observed that mutants in the MAPK pathway suppressed the reduction in worm survival, emphasizing its role in survivability, meiotic progression, and DNA damage checkpoint-mediated apoptosis. This study reveals complex interactions between herbal extracts and C. elegans processes, shedding light on potential antitumor effects and mechanisms. The findings provide insights into the complex landscape of herbal medicine's impact on a model organism, offering implications for broader applications.

We investigate some information properties of consecutive k-out-of-n:G systems in light of fractional generalized cumulative residual entropy. We firstly derive a formula to compute fractional generalized cumulative residual entropy related to the system's lifetime and explore its preservation properties in terms of established stochastic orders. Additionally, we obtain useful bounds. To aid practical applications, we propose two non parametric estimators for the fractional generalized cumulative residual entropy in these systems. The efficiency and performance of these estimators are illustrated using simulated and real datasets.

Lithraea molleoides fruit gum (LMFG) is a valuable product obtained from the total hydrolysis of the fruit. The hydrolysis process involves three methods: thermal (LMFGT), basic (LMFGB), and acid (LMFGA). Through these methods, the aim is to break bonds and de-esterify polysaccharides, resulting in increased solubility and decreased molecular weight. The resulting hydrolysates are then combined with pectins in a 1:2 ratios to form films. In this study, the focus is on utilizing the hydrolysates of Lithraea molleoides gums for film applications, with an evaluation of their structural and physicochemical characteristics. The films produced exhibit excellent mechanical properties and low water vapor permeability, as well as exceptional thermal stability. These properties make them highly suitable for industrial films in pharmaceutical and food applications. This research highlights the potential of LMFG-based films as a viable solution for various industrial needs due to their outstanding performance across multiple parameters.

Fall armyworm (FAW), Spodoptera frugiperda, has been causing significant crop losses and has now become the most economically important pest of corn globally. With the challenges arising from the extensive use of synthetic chemicals such as reports that FAW gained resistance to various insecticides, researchers are developing environmentally sound control measures to address these issues. An example is the application of the RNA interference in managing FAW populations. RNA interference is a biological process where dsRNA molecules silence specific gene of interest. Here, dsRNA fragments were generated to target Cytochrome P450 monooxygenases CYP321A7 and CYP321A8 in FAW. Mortality and phenotypic effects of RNAi-mediated knockdown of essential genes of target CYP450s through RNAi in FAW were analyzed. Results showed that RNA in vitro transcription yielded ~14 μg/μL of dsCYP321A7 and dsCYP321A8. The mortality rate of dsCYP321A7- and dsCYP321A8-fed larvae following exposure to insecticide with dual active ingredient of Chlorantraniliprole and Thiamethoxam increased by 90% and 80%, respectively as compared to dsGFP-fed larvae with 40% at 2 days post insecticide exposure. This study provided evidence for the potential of RNAi for FAW management strategies particularly for insecticide-resistant populations, including the potential for biopesticide development.

We investigated the potential of Mucuna pruriens and Canavalia ensiformis as phytoremediation candidates for Tebuthiuron (TBT)-contaminated soils. Crotalaria juncea was the bioindicator specie for tebuthiuron and Lactuca sativa used in ecotoxicological bioassays, both analyses aim to verify the herbicide presence in soil samples as TBT is persistent with environmental and health risks due to its tendency to bioaccumulate. We conducted a 140-day greenhouse experiment associated with bioaugmentation by microbial inoculants - solid or liquid. Both C. ensiformis (CE) and M. pruriens (MP) exhibited distinct growth patterns, with MP demonstrating a faster growth rate than CE, evident from its higher growth apex (α = 84.52) and specific growth rate (k = 0.113). While soil samples with microbial inoculants demonstrated significant biomass increase with tebuthiuron absence, the introduction this herbicide notably hindered phytomass production, particularly in MP + TBT treatments. Liquid inoculant notably contributed to biomass accumulation in MP, whereas microbial bioaugmentation did not substantially enhance phytomass in CE under tebuthiuron presence. Furthermore, C. juncea exhibited initially slow development in the reference treatment but displayed accelerated growth and greater height with liquid inoculant compared to solid inoculant. Despite the presence of tebuthiuron, C. juncea in soil with TBT alone reached a maximum height exceeding 100 cm. However, tebuthiuron significantly reduced biomass accumulation in C. juncea, indicating its high sensitivity to this herbicide. Liquid inoculant facilitated the highest dry biomass accumulation in C. juncea, yet the introduction of tebuthiuron led to a considerable reduction in biomass, underscoring the plant's susceptibility to it. While bioaugmentation showed potential in mitigating herbicide phytotoxicity, its efficacy varied among tested inoculants. In bioassays with L. sativa, bioaugmentation with liquid inoculant resulted in a higher final germination index (GI) compared to solid inoculant and the reference treatment. Conversely, soil samples containing only tebuthiuron demonstrated a higher final GI and growth velocity compared to the reference treatment, implying the natural attenuation of toxicity over time. Additionally, previous cultivation of Mucuna pruriens positively influenced the development of test-organism, resulting in a higher GI and growth rate compared to the reference treatment. These findings provide valuable insights for various stakeholders, including the scientific community, industries, and bioengineers. They offer a strong incentive to develop advanced bioremediation strategies to address persistent pesticide contamination, especially in intensive agroecosystems like sugarcane-producing fields. Implementing our biological solution holds the potential to usher in a cleaner and safer era in agriculture, thereby protecting both society and the environment.

Porphyromonas gingivalis is the major pathogenic bacteria found in the subgingival plaque of patients with periodontitis, leading to neuroinflammation. The bacteria destroy periodontal tissue through its virulence factors which are retained in the bacteria’s outer membrane vesicles (OMV). The purpose of this study was to determine the real time effect of intraperitoneal in-jection of P. gingivalis OMV on the production and expression of inflammatory markers as well as histopathological changes in the central nervous system (CNS) of adult zebrafish. Fol-lowing the LD50 (21 µg of OMV), the zebrafish were injected intraperitoneally with 18 µg of OMV and control group with normal saline at 7 different time points. Brains of experimental zebrafish were dissected at desired time points for colorimetric assays, ELISA and histology. This study discovered that nitric oxide and PGE2 were significantly increased at 45 minutes while IL-1β and IL-6 were expressed at subsequent 12- and 24-hours. respectively. Histopatho-logical changes such as blood coagulation, astrocytosis, oedema, spongiosis, and necrosis were also observed between 6- and 24-hours. The two apoptotic enzymes, caspases 3 and 9 were not expressed at any time point. In summary, the OMV induced neuroinflammatory responses and histopathological changes in adult zebrafish were time point dependent

Balanites aegyptiaca del. (Zygophyllaceae) is widely used in traditional medicine, both human and veterinary, throughout Africa for its many properties, including antiparasitic properties. This experimental study aims to optimise the extraction conditions of the seeds of Balanites aegyptiaca del. Aqueous maceration was carried out with mass to volume ratios of 40%, 30%, 20%, 10% and 5% and extraction times of 6, 12, 24, 36 and 48 hours. Extraction yields, phytochemical screening, saponins assay, antioxidant activities (ABTS and FRAP) and antiparasitic tests on Heligmosomoides bakeri were used to compare the different extracts. The pharmaco-chemical study generally showed that aqueous maceration gave the best results, with a mass/volume ratio of 10% after 12 hours of maceration. The yield obtained was 28.03% with a saponins content of 13.81 mg/g. The antioxidant activities were 4.25 ± 0.17µg/mL by the ABTS method and 0.739µg/mL by the FRAP method. The larvicidal activity also showed that the 10%12h extract produced 100% larval mortality from 25ug/mL. These data provide a basis for guiding the choice of extraction process parameters in producing this antiparasitic phytomedicine.

This study was carried out with a low-cost electronic nose prototype based on eight metal oxide sensors (MQ) in order to characterize samples of lemons treated with 0.5% and 0.1% of sodium benzoate. The MQ sensors designed are sensitive to one or more chemicals to detect the presence of a variety of chemicals in the air. The sensor MQ135 detects ammonia, hydrogen sulphide and benzene. Signal data were studied to obtain a pattern recognition of rotten in lemon fruits. Network analysis was used to obtain a calibration of measures among the stage of lemons. In this article, an electronic nose prototype based on 8 MQ metal oxide sensors has been used in order to analyze and characterize different lemon varieties to which different chemical treatments have been applied in pre-harvest. PCA-based data analyzes were used to observe clusters in the data. Through the combined use of the data obtained by the nose and these Sequential Neural Networks (SNNs) a classification tool for lemon varieties and applied treatments has been obtained. It is shown the ability of this device to be used as a reliable discrimination method, in addition to providing low cost and optimization of time and expert resources.

Background: Across the world, antibiotic resistance is a grave problem. To address this issue, it is necessary to investigate new antibiotic sources. Aim of the study: Egypt's diverse soil habitats were used to produce bacterial Myxopyronin B, which was then tested for antimicrobial activity in preclinical animal studies and randomized human clinical trials stages 1/2. Type of the study: Screening experimental study. Methodology: Egypt's various soil conditions were examined for the development of bacterial isolates that produced the antibiotic compound Myxopyronin B. Reversed phase HPLC was used to purify Myxopyronin B. To determine the test antibiotic's minimum inhibitory concentration( MIC) and invitro antibacterial activity, the Broth microdilution method and the Paper disc diffusion assay were utilized. Moreover, in vivo antibacterial spectrum, adverse medication responses, and pharmacokinetics were found in preclinical animal testing stages and phases 1/2 of randomized clinical studies involving volunteer humans. Results: Myxopyronin B was generated from the culture supernatant of Myxococcus SDU36, the main soil bacterial isolate cultured on a Casein yeast peptone plate. With MICs less than 100 mcg/ml, the test antibiotic prevented the growth of several Gram +ve bacteria, whereas, at MICs higher than 100 mcg/ml, it inhibited the development of a small number of Gram -ve bacteria, including Escherichia coli. However, eukaryotic cells—such as those found in fungi and humans—were unaffected. The test antibiotic was seen to inhibit prokaryotic DNA-dependent RNA polymerase( RNLP), indicating its bactericidal activity. Mean Cmax was 9-10 mcg/ ml at mean Tmax 1 hour when 600 mg dose was orally administered in randomized human clinical trials phases 1/2, and T1/2 reached 2.25 hours following first-order kinetics of elimination. Duration of its action was nearly 8 hours after oral administration. Rare toxicity was detected during preclinical and randomized human clinical trials phases 1/2 in the form of mild diarrhea and GI upset in less than 7 % of experimental candidates. It exhibited approximately 90% plasma protein binding, especially with albumin which resulted in prolonged therapeutic action. It showed a concentration dependant antibiotic killing effect. With the concentration-dependent killing pattern, the higher the drug concentration relative to the pathogen minimum inhibitory concentration( MIC), the greater the rate and extent of antimicrobial activity. Conclusion: The present study was promising due to the production of the bactericidal antibiotic Myxopyronin B from Myxococcus sp. SDU36 isolated from different soil environments in Egypt.

Background: Across the world, antibiotic resistance is a critical challenge. To address such a problem, it has to be done to investigate new antibiotic sources. Aim of the study: Study of the antibacterial activity of Corallopyronin A in preclinical animal testing and randomized human clinical trials stages 1/2, as well as investigating the purification of Corallopyronin A from different soil conditions in Egypt. Type of the study: Screening experimental study. Methodology: Egypt's various soil conditions were tested to develop bacterial isolates that produced the antibiotic compound Corallopyronin A. Reversed phase HPLC was used to purify Myxopyronin A. The broth microdilution method and the paper disc diffusion assay determined the test antibiotic's minimum inhibitory concentration( MIC) and in vitro antibacterial activity. Moreover, in vivo antibacterial spectrum, adverse medication responses, and pharmacokinetics were found in phases 1/2 of randomized clinical studies involving humans and animal models. Results: Corallopyronin A was generated from the culture supernatant of the soil bacterial isolate Corallococcus coralloides M2, cultivated on a Casein yeast peptone( CYP) plate. The test antibiotic prevented the growth of several Gram-ve bacteria, including Escherichia coli, at MICs higher than 100 mcg/ ml; while blocking the development of many Gram +ve bacteria, with MICs ranging from 3 to 15 mcg/ ml. However, eukaryotic cells—such as those found in fungi and humans—were unaffected. A bactericidal consequence of the test antibiotic was detected in the inhibition of prokaryotic DNA-dependent RNA polymerase( RNLP). Conclusion: The synthesis of the bactericidal antibiotic Corallopyronin A from Corallococcus coralloides M2 isolated from different soil settings in Egypt intrigued the current investigation.

Visceral leishmaniasis is a disease caused by protozoa of the species Leishmania (Leishmania) infantum (syn=Leishmania chagasi) and Leishmania (Leishmania) donovani, which are transmitted by hematophagous in-sects of the genera Lutzomyia and Phlebotomus. The domestic dog (Canis familiaris) is considered the main ur-ban reservoir of the parasite due to the high parasite load on its skin, serving as a source of infection for sand-fly vectors and, consequently, perpetuating the disease in the urban environment. Some factors are consid-ered important in the perpetuation and spread of canine visceral leishmaniasis (CVL) in urban areas, such as stray dogs with their errant behavior and houses that have backyards with trees, shade, and organic materi-als, creating an attractive environment for sandfly vectors. CVL is found in approximately 50 countries with the number of infected dogs reaching millions and, due to the difficulty of controlling and diagnosing the disease, the number of infected animals could be even greater. In the four continents endemic for CVL there are reports of disease expansion in endemic countries such as Brazil, Italy, Morocco, and Tunisia, as well as in areas where CVL is not endemic, for example, Uruguay. Socio-environmental factors, such as migration, drought, deforestation, and global warming, have been pointed out as one of the reasons for expansion to ar-eas where it had been absent. Thus, the objective of this review is to address (i) the distribution of CVL in endemic areas (ii) the role of the dog in the visceral leishmaniasis epidemiology and the factors that influence dog infection and spread of the disease, and (iii) the challenges faced in the control of CVL.

Rhodotorula mucilaginosa fungaemia is rare and highly resistance to antifungal therapy. We herein report a case involving a 31-year-old male admitted after a high-velocity road traffic accident. He sustained a grade IV liver injury with right hepatic vein thrombosis, necessitated an urgent laparotomy. Post-operatively, repeated imaging of the abdomen revealed presence of biloma. Percutaneous subdiaphragmatic drainage was done but appeared ineffective, prompting second surgery for a urgent hemi-hepatectomy. Patient then was nursed in the intensive care unit (ICU) however during his stay in the ICU he became more sepsis evident by worsening ventilatory support and rise in septic parameters from the biochemistry parameters. Despite on intravenous piperacillin-tazobactam and fluconazole, his septic parameters did not improve and a full septic workup was done and was found to be positive for Rhodotorula mucilaginosa from the blood cultures. After discussion with the infectious disease physicians and clinical microbiologist, it was decided to initiate a course of intravenous meropenem and amphotericin B based on minimum inhibitory concentration (MIC) values, considering the patient's extended ICU stay and catheter use. Eventually, successful weaning off mechanical ventilation was discharge from ICU care. This case underscores the necessity of individualized approaches, combining timely imaging, appropriate drainage techniques, and tailored treatments to optimize outcomes for such intricate post-traumatic complications.

Several bio-actors are involved in the occurrence of leishmaniasis, which is made too complex for prevention and control options. Currently, all forms of leishmaniasis are being treated with chemical drugs, which have limitations and adverse effects on patients. Searching the bioactive molecules from plant sources allows the development of a novel drug. The essential oil and solvent extract of the roots of E. kebericho showed antileishmanial activity. But looking for biological activities of the isolated active ingredient from tubers of E. kebericho was scant. Thus, the isolation of the leishmanicidal compound from the roots of E. kebericho through a bioassay-guided technique was an aim of this work. Dehydrocostus lactone showed doubled potent than the standard drug miltefosine on leishmanicidal activities against L. major and L. tropica. Besides, the broad-spectrum biological activities of dehydrocostus lactone could be a potential drug candidate for co-infected patients caused by an association of leishmania and another pathogen.

Salmonella typhimurium is the predominant serovar identified in cases of salmonellosis. It is recognised for adhering to various surfaces and creating biofilm, leading to several foodborne outbreaks and antibiotic resistance. Actinomycetes are thought to possess a wide array of antibacterial characteristics that can combat different diseases. Currently, it is unclear whether it has antibiofilm properties against S. typhimurium. This study aimed to assess the antibiofilm efficacy of actinomycetes extracts against S. typhimurium ATCC 14028 by the Crystal Violet assay. Four actinomycetes isolates, EAS5, EAS7, EAA141, and EAA11, were obtained from BRIS soil in Terengganu. The actinomycetes isolates were extracted using ethyl acetate (EA). S. typhimurium was exposed to extracts for 6, 12, and 18 hours. The results showed that all EA extracts effectively prevented the S. typhimurium biofilm at 12 and 18 hours. EAAA11 was the most effective against the 12-hour biofilm, whereas EAS5 was the most effective against the 18-hour biofilm among these extracts. In conclusion, actinomycetes isolated from the BRIS soil in Terengganu show potential for controlling Salmonella infection.

Saliva, as a diagnostic medium, offers a promising alternative to blood by virtue of its non-invasive collection, which enhances patient compliance, especially in paediatric and geriatric populations. In this study, we assessed the utility of saliva as a non-invasive medium for measuring V. cholerae-specific serum antibodies in naturally infected individuals. We tested paired serum and saliva samples obtained from a total of 63 cholera patients enrolled in a cohort study. Vibriocidal antibodies assay (IgM/IgG) as markers for accurate determination was used to determine cholera specific antibody levels. Using receiver operating characteristics (ROC) curve, we found that the best cut-off that maximizes (sensitivity + specificity) is 10 titers. At this saliva titer, the sensitivity is 76.9% (95%CI: 60.9%, 87.7%) and specificity is 80.0% (95%CI: 56.6%, 92.5%). Using Spearman’s correlation coefficient, we also found evidence of a positive correlation between Vibrio Cholerae saliva and serum antibodies (rho=0.66, p&lt;0.001). In conclusion, saliva-based diagnostic cholera tests has high diagnostic accuracy, and would be advantageous, cheaper, and quicker for early diagnosis of severe cholera outcomes.

Barcoding of DNA is a species identification technique that utilizes a small DNA fragment from a specific gene or genes known as "sequencing." The idea behind DNA barcoding is that a single sequence can be used to specifically identify an organism to species by comparison with a reference library of these DNA sections. Herbal adulteration can be monitored using DNA barcodes that utilize medicinal plants such as Atropa belladonna. Because it is challenging to distinguish between species solely based on morphology, other plants belonging to the same genus are replacing it in trade. Such replacement with plants not utilized for medical purposes could have negative side effects. Also, Atropa belladonna's positive effects can be completely lost. Taxonomists also have trouble distinguishing the genuine Atropa belladonna plant from cultivated items. As a result, DNA barcoding can now be used to overcome this challenge. The development of DNA barcodes may help with trade control, which protects and conserves endangered species from export outside of the country due to their economic and biological significance. In this review, we discuss the DNA barcoding of Atropa belladonna plant in the medicinal plants field.

: Background: The polysaccharides are one of the main active ingredients of Polygonum sibiricum (PSP), which is a traditional food and medicine homologues in China. The antidepressant-like effects of PSP and its underlying mechanisms remain to be studied in depth, especially the regulation of microglial polarization. Methods: The chemical composition and structural characteristics of PSP was determined firstly. The CUMS procedure was carried out to the zebrafish for 5 weeks and PSP were immersed administration for 9 days (1 h/d). The body weight of zebrafish was monitored and the behavioral tests, including the novel tank test and light and dark tank test, were performed to evaluate the antidepressant-like effects of PSP. Then the function of HPI axis, the levels of peripheral inflammation, neuronal and blood-brain barrier damage in mesencephalon and telencephalon and the mRNA expression of M1/M2 phenotype genes in brain were examined. Results: PSP samples had the typical structure characteristics of polysaccharide, consisting of glucose, mannose and galactose with the average Mw 20.48 kDa, which owned a porous and agglomerated morphology. Compared with untreated zebrafish, the depression-like behaviors of CUMS-induced zebrafish were significantly attenuated. PSP significantly decreased the levels of cortisol and the pro-inflammatory cytokines and increased the levels of the anti-inflammatory cytokines in the body of CUMS-induced depressive zebrafish. Furthermore, neuronal and blood-brain barrier damage in mesencephalon and telencephalon and the mRNA expression of M1/M2 phenotype genes in brain were remarkably reversed by PSP. Conclusion: These findings indicated that the antidepressant-like effects of PSP were related to alter the HPI axis hyperactivation, suppress peripheral inflammation, inhibit neuroinflammation induced by microglia hyperactivation, and modulate microglial M1/M2 polarization. The current study provides foundation to the future exploitation of PSP in the functional foods of emotional regulation.

Biocides used in antifouling (AF) paints, such as 4,5-dichlorine-2-n-octyl-4-isothiazole-3-one (DCOIT), can gradually leach into the environment. Some AF compounds can persist in the marine environment and cause harmful effects on non-target organisms. Nanoengineered materials, such as mesoporous silica nanocapsules (SiNC) containing AF compounds, have been developed to control their release rate and reduce their toxicity to aquatic organisms. This study aimed to evaluate the acute toxicity of new nanoengineered materials, SiNC-DCOIT and a silver-coated form (SiNC-DCOIT-Ag), as well as the free form of DCOIT and empty nanocapsules (SiNC), on the sun coral Tubastraea coccinea. T. coccinea is an invasive species and can be an alternative test organism for evaluating the risks to native species, as most native corals are currently threatened. The colonies were collected from the Alcatrazes Archipelago, SP, Brazil, and acclimatized to laboratory conditions. They were exposed for 96h to different concentrations of the tested substances: 3.33, 10, 33, and 100 µg L-1 of free DCOIT; 500, 1000, 2000, and 4000 µg L-1 of SiNC; and 74.1, 222.2, 666.7, and 2000 µg L-1 of SiNC-DCOIT and SiNC-DCOIT-Ag. The test chambers consisted of 500 mL flasks containing the test solutions, and the tests were maintained under constant aeration, constant temperature of 23 ± 2°C, and photoperiod of 12h:12h (light/dark). At the end of the experiments, no lethal effect was observed; however, some sublethal effects were noticeable, such as the exposure of the skeleton in most of the concentrations and replicates, except for the controls, and embrittlement at higher concentrations. Adults of T. coccinea were considered little sensitive to the tested substances. This resistance may indicate a greater capacity for proliferation of the species, which is favored in substrates containing antifouling paints, to the detriment of the native species.

This study carried out a qualitative and descriptive relational approach that combined Documentary Analysis and elements of the Analytical Method to investigate the phenomenon of bilingualism in Tuluá, Colombia. The aim of this study was to diagnose the promotion of bilingualism by proposing a series of recommendations to strengthen the educational and linguistic policies. The data was processed through matrices and analyzed in the light of the theoretical references to enable a historical account of the programs or strategies that have been implemented for the promotion of bilingualism. The findings of this study are crucial in developing suggestions for enhancing the effectiveness of bilingualism. By identifying strengths, weaknesses, and patterns in the historical framework and modern performance indicators, the research offers valuable contributions to the current discourse on bilingual education policies in Tuluá, Colombia.

Silybum marianum is a therapeutic plant belonging to the family Asteraceae. It contains silymarin, as its principal component, and has been reported to demonstrate hepatoprotective activity. The study was aimed to investigate the phytochemical constituents, synthesis of nanoparticles, antioxidant activity and antimicrobial effects of the leaf extract. Extracts were synthesized from AgNO3 and characterized via FT-IR, U.V and XRD. The antioxidant activity was determined using DPPH, H2O2 and β-carotene assays. Similarly, all extracts expressed significant DPPH inhibition (IC50) of 5.1, 5.0, 4.7, 2.3, 4.3 and 1.1 (ug/mL), H2O2 of 1.3, 2.2, 4.7, 5.6, 4.0 and 1.0 (ug/mL) from the methanol/water extract, nanoparticles, n-Hexane, ETOAC, n-Butanol and Ascorbic acid (standard) respectively. The extract exerted scavenging activity on β-carotene with a significant IC50 inhibition of 1.5, 0.8, 5.2, 0.8, 2.7, 4.8 at 0 hour and 1.5, 0.8, 5.2, 0.8, 2.7 and 4.8 (ug/mL) after 2 hours water bath incubation at 50°C in the order of the extracts above. Additionally, nanoparticles and plant extracts significantly (p<0.0001) inhibited the growth of bacteria with mean zone of inhibition ranging 7-13 mm and 4-11 mm. This study demonstrated that extracts from different solvents of S. marianum possess significant antioxidant activity, as well as antimicrobial effects which are all concentration dependent. Thus, the result suggests that methanolic/water leaf extract of S. marianum could serve as a potential source of antioxidants and can be explored as a therapeutic agent in free radical induced diseases.

Since the early 2000s, high-risk E. coli clones that presented a highly virulent and multidrug resistance profile have been identified, and their genome is completely sequenced. Molecular techniques make it possible to follow the evolution and spread of those clones in different parts of the world. The results of several studies monitoring the emergence of new high-risk E. coli clones showed an increase in the number of clones and the virulence strategies and in the emergence of clones with the ability to cause intestinal and urinary infections combined with a multidrug resistance profile. We certainly can't stop the emergence of new high-risk clones. The remaining questions are: How can we use the knowledge about E. coli high-risk clones to our advantage in the war against multidrug antibiotic resistance? How can we stop the spread of those clones and improve diagnostic and treatment stewardship? This is a challenge that requires collaboration and shared knowledge.

Thurstonian scaling, i.e., Thurstonian discriminal distance δ or d' can be used as a sensory measurement index to measure and monitor food sensory difference/similarity between test and control samples due to potential food contamination. It can be obtained from any one of the sensory discrimination methods. Thurstonian scaling is theoretically independent of methods or scales used for its estimation. This paper discusses statistical inference including estimations and tests of hypothesis for d'. Ten basic sensory discrimination methods including 6 forced-choice methods and 4 methods with response bias are used in this paper to estimate d' values and their variances. Statistical tests are conducted based on the estimated d' values and their variances. The statistical tests include difference testing and equivalence/similarity testing for individual d' values for test and control samples and for two or multiple d' values for test samples. Application and significance of Thurstonian scaling for sensory discrimination methods is discussed generally. R codes for estimations and tests for d' values are provided in the paper.

Background The use of antimicrobials to treat food animals leaves antimicrobial residues in foodstuffs. The World Health Organization (WHO) defines the acceptable daily intakes (ADIs) of these residues as the dose of these antimicrobials that is safe for an average human to consume on a daily basis. We hypothesized that the low dose of ciprofloxacin classified as safe by the WHO could select for ciprofloxacin resistant strains of Klebsiella pneumoniae in a Galleria mellonella model. Objectives We aimed to evaluate if the consumption of peri-ADI doses of ciprofloxacin could select for ciprofloxacin-resistant (Ser464Phe, GyrB, ciprofloxacin MIC of 4 µg/mL) compared to -susceptible (isogenic, ciprofloxacin MIC of 0,047 µg/mL) strains of K. pneumoniae in a Galleria mellonella model. Methods A model of K. pneumoniae infection in G. mellonella larvae was used for the experiment. The larvae were inoculated with K. pneumoniae followed by 10x ADI, 1x ADI, 1/10^th ADI, 1/100^th ADI, 1/1000^th ADI doses of ciprofloxacin. Isolation of K. pneumoniae colonies was then performed on selective agar plates with and without ciprofloxacin (1 µg/mL) plates. The proportion of colonies with ciprofloxacin resistance was then calculated for each group at 24 and 48 hours. Results A significant increase was seen in the proportion of resistance for the 1x ADI and 1/10^th ADI concentration on day 2 compared to the positive control. Conclusion We found that at 48 hours, there was an enrichment of K. pneumoniae colonies with ciprofloxacin resistance in the larvae receiving 1x ADI and 1/10^th ADI concentrations of ciprofloxacin. These results suggest that the ciprofloxacin MSC_select for K. pneumoniae in this model is 1/10^th of the acceptable daily concentration (ADI) dose of ciprofloxacin, which is equivalent to 0.239 ng/µl.

Peroxisome proliferator-activated receptors (PPARs) are important targets for cannabidiol (CBD), which mediate many of its biological actions. The hypothesis of the present research assumed that PPARs affect the gut response to different challenge factors in chickens (C. perfringens vs lipopolysaccharides (LPS) from E. coli), and CBD can mediate the pathways of this response. The study proved that CBD and challenge factors significantly affect the expression level of PPARα (P=0.001) and selected genes determining gut barrier function. A positive correlation was demonstrated between PPARs and genes involved in the formation of tight junctions, immune and oxidative stress responses in chickens. Dietary supplementation with CBD actively mediated the expression rate of PPARs, but the mechanism of interaction between CBD and PPARs was different depending on the stress factor used. The addition of CBD to the birds’ diets did not contribute to reducing intestinal permeability under induced stress conditions and did not cause a stress-inducing effect resulting in the increase of cortisol and endotoxin concentration in the blood. CBD also supported the mechanisms of protecting intestinal cells from cytotoxic effects in a C. perfringens challenge. This study indicates the importance of research towards understanding the mechanisms of PPARs as a target for enhancing intestinal barrier function, provides new results on the biological action of CBD in chickens and shows a constant PPARs association with the jejunum mucosa of birds.

This article presents the results of isolating and identifying EPF species and screening thermotolerant and highly virulent EPF strains that are promising for controlling Colorado potato beetle populations. Soil samples were collected from southern Kazakhstan for the isolation of fungi used in the experiments. Using the insect bait method, 41 isolates of EPF were obtained from the soils of southern Kazakhstan. PCR analysis based on ITS and TEF loci (elongation factor Ef1a) showed that all the isolates belong to the species B. bassiana. However, analysis of the ITS locus revealed two cryptic species: B. bassiana and B. pseudobassiana. No other fungal species were detected, likely due to the insufficient informativeness of the ITS and TEF loci in distinguishing species. Further studies with other loci are required. As a result of selecting natural isolates for virulence and thermotolerance, the following strains were identified: B13, B14, B15, B22, B23, B24, B25, B23-23, Bc4, Bc7, and Bc8. These strains showed high virulence (100% on the 11th day after treatment) and tolerance to suboptimal temperatures (+15°C and +30°C). Therefore, they are considered the most promising for use in the context of climate change. The data suggest that, despite the source of the isolates, EPF have great potential for controlling Colorado potato beetle populations, as the majority of isolates lead to fatal outcomes. Their geographical origin primarily influences their temperature preferences, which negatively affect their effectiveness in field conditions.

Onychomycosis is a common fungal nail infection for which new antibiotics are needed to overcome antifungal resistance and other limitations of conventional treatments. This study reports the development of antifungal nail lacquers containing oregano essential oil (OEO), rosemary essential oil (REO), and/or biogenic silver nanoparticles (bioAgNP). The formulations (F) were tested against dermatophytes by agar diffusion, ex vivo nail infection, and scanning electron microscopy techniques. They were evaluated pharmacotechnically and by FTIR-PAS to access permeation across the nail plate. F-OEO and F-OEO/Ag were promising for the final nail lacquer formulation, they permeated through the nail plate and showed antifungal efficacy against dermatophytes-contaminated nails after 5 days of treatment. The treated nails had decreased hyphae and spores compared to untreated control; the hyphae were atypically flattened which indicates loss of cytoplasmic content due to damage to cytoplasmic membrane. Formulations were stable after centrifugation and thermal stress, maintaining organoleptic and physicochemical characteristics. F-OEO and F-OEO/Ag had pH compatible with the nail, and drying time (59-90 s) within the reference for nail lacquer. For the first time, OEO and bioAgNP were incorporated in a nail lacquer, resulting in a natural and nanotechnological antifungal product for onychomycosis that may combat microbial resistance.

- Early and accurate detection of pneumonia from chest X-ray images is crucial for timely treatment and patient care. In study presents a robust computational framework leveraging the CheXnet algorithm, a deep learning model, to identify pneumonia from chest X-rays. Developed a neural network architecture trained on a substantial dataset of chest X- ray images labeled as 'Normal' and 'Pneumonia'. The user- friendly interface of the system allows for seamless uploading of images, with the model subsequently providing predictions, a confusion matrix, accuracy metrics, and layer activations visualizations. The algorithm's performance was rigorously evaluated, with a confusion matrix indicating a high number of true positives and true negatives. Specifically, the model detected pneumonia with an accuracy of 92.47%, as shown by the results displayed in the system's interface. The precision, recall, and F1-score for pneumonia detection were 0.90, 0.96, and 0.93 respectively, with the overall system achieving an accuracy of 0.91 across the test set of 624 images. Visualizations of the neural network layers reveal the complex feature mappings the model uses to differentiate between normal and pathological findings. These insights could be vital for enhancing the interpretability of deep learning models in medical diagnostics. The Results indicate that the CheXnet-based neural network is a promising tool for automating pneumonia detection in clinical settings, which can assist radiologists in the diagnostic process and potentially improve patient outcomes through earlier intervention.

In the introductory part of this review, the relevant results of mostly the latest experimental studies on carotenoid absorption and availability in the human body are briefly presented. The subsequent parts summarise such topics as carotenoid carriers, body transport and tissue delivery, before finally reporting on molecular-level studies of carotenoid binding by membrane receptors. The main message of the review is contained in the section describing computational investigations of carotenoid intercalation and dynamic behaviour in lipid bilayers. The relevance of these computational studies lies in showing the direct link between the microscopic behaviour of molecules and the characteristics of their macroscopic ensembles. Furthermore, studying the interactions between carotenoids and lipid bilayers, and certainly proteins, on the molecular- and atomic-level using computational methods facilitates interpretation and explanation of the macroscopic properties of these systems and, hopefully, helps to better understand the biological functions of carotenoids.

This study utilized phytochemical screening to conduct qualitative analysis of plant extracts, aiming to identify various classes of secondary metabolites. Moreover, the antibacterial activity of different types of O. vulgare and S. triloba extracts was determined. To achieve the aim of the study, aqueous, ethanolic, and enzymatic extracts were prepared and screened for phytochemical capacity and antioxidant activities. Determination of the antibacterial activity included phenotypic screening of antibiotic susceptibility pattern of oral and food pathogenic bacterial strains, determination of Minimum inhibitory concentration and minimum bactericidal concentration - Microdilution broth test and in vitro valuation of antibacterial efficacies - anti-biofilm properties of the studied herbal extractions. Results: Our study evaluated the phytochemical composition and the antioxidant, antibacterial, and anti-biofilm properties of O. vulgare and S. triloba extracts. The analyzed samples contained bioactive compounds, such as phenolics and flavonoids, contributing to the observed strong antioxidant effect. Furthermore, they exhibited notable activity against oral biofilm formation and demonstrated significant antibacterial efficacy against dental caries microorganisms as well as food pathogens. Despite methodological variations, all extracts showed significant antioxidant capacity and promising antibacterial activity against various pathogens, including resistant strains, while also inhibiting biofilm formation. Although limited to two plant species and facing methodological constraints, the study lays a groundwork for future research, indicating the therapeutic potential of O. vulgare and S. triloba extracts. Further exploration is needed to report on underlying mechanisms and validate efficacy through clinical trials.

Tomato is one of the most important vegetable crops grown worldwide. Tomato brown rugose fruit virus (ToBRFV), a seed-borne tobamovirus, poses a serious threat to tomato productions due to its ability to break the resistant genes (Tm-1, Tm-2, Tm-22) in tomato. The objective of this work was to identify new resistance source(s) of tomato germplasm against ToBRFV. To achieve this aim, a total of 476 accessions from 12 Solanum species were tested with the ToBRFV US isolate for their resistance and susceptibility. As a result, a total of 44 asymptomatic accessions were identified as resistance/tolerance, including 31 accessions of S. pimpinellifolium, one accession of S. corneliomulleri, four accessions of S. habrochaites, three accessions of S. peruvianum and five accessions of S. subsection lycopersicon hybrid. Further analysis using serological tests identified four highly resistant S. pimpinellifolium lines, PI 390713, PI 390714, PI 390716 and PI 390717. The inheritance of resistance in the selected lines was verified in next generation and confirmed by RT-qPCR. To our knowledge, this is a first report of high resistance to ToBRFV in S. pimpinellifolium. These new genetic resources will expand the genetic pool available for breeders to develop new resistant cultivars of tomato against ToBRFV.

Tobamovirus species represent a threat to solanaceous crops worldwide, due to their extreme stability and being seed-borne. In particular, recent outbreaks of tomato brown rugose fruit virus in tomato and pepper crops led to the establishment of prompt control measures, and the need for reliable diagnosis was urged. Another member of the genus, tomato mottle mosaic virus, has recently risen attention due to reports in different continents and its common features with tomato brown rugose fruit virus. In this study, a new real-time RT-PCR detection system was developed for tomato brown rugose fruit virus and tomato mottle mosaic virus on tomato leaves and seeds using TaqMan chemistry. This test was designed to detect tomato mottle mosaic virus by amplifying the movement protein gene in a duplex assay with tomato brown rugose fruit virus target on the CP-3’NTR region, which was already validated as a single assay. The performance of this tests was evaluated, displaying analytical sensitivity 10-5-10-6-fold dilution for seeds and leaves, respectively, and good analytical specificity, repeatability, and reproducibility. Using the newly developed and validated test, tomato brown rugose fruit virus detection was 100% concordant with previously performed analyses on 106 official samples collected in 2021 from different continents.

Trichomes are a common morphological defense against pests, in particular, type IV glandular trichomes have been associated with resistance against different invertebrates. Cultivated tomatoes usually lack or have a very low density of type IV trichomes. Thus, specific breeding programs to incorporate these natural defences, that are common within the Solanum genus, might improve a more sustainable management. We have identified a S. pimpinellifolium accession with very high density of this type of trichomes. Two F2 mapping populations using two different parents have been developed, characterized and genotyped using a new genotype methodology, K-seq. We have been able to build an ultra-dense genetic map with 147,326 markers with an average distance between markers of 0.2 cM that has allowed us to perform a detailed mapping. We have used two different families and two different approaches, QTL mapping and QTL-seq, to identify several QTLs implicated in the control of trichome type IV developed in this accession on the chromosomes 5, 6, 9 and 11. The QTL located on chromosome 9 is a major QTL that has not been previously reported in S. pimpinellifolium that increases by a factor of 9 the density of trichomes.

The Maastricht VI/Florence consensus recommends, as one of the measures to enhance the efficacy of Helicobacter pylori (H. pylori) infection eradication, a personalized treatment approach involving the selection of an antimicrobial agent based on the pre-determined resistance of H. pylori. To meet the need to develop test systems for personalized drug selection, we conducted a study of H. pylori molecular resistance in Moscow using a self-developed Sanger sequencing test platform. We validated our test system on 25 pure culture samples of H. pylori with known resistance. Sensitivity and specificity for detecting resistance to clarithromycin was 100%, and to levofloxacin - 93% and 92%, respectively. Then we tested our platform on 112 H. pylori -positive patients who had not previously received proton pump inhibitors (PPI) or antibacterial drugs. Mutations indicating resistance to clarithromycin were found in 27 (24%) samples, and to levofloxacin in 26 (23%) samples. Double resistance was observed in 16 (14%) samples. The most common mutations leading to clarithromycin resistance were 2143G and 2142G, to levofloxacin resistance – 261A and 271A in the gyrA gene, which account for 69% of all identified genetic determinants in levofloxacin-resistant bacteria. Thus, a personalized approach to the selection of H. pylori eradication therapy based on the detection of bacterial resistance before prescribing first-line therapy could help avoid the prescription of ineffective H. pylori eradication therapies and, overall, contribute to the control of antibiotic resistance of H. pylori.

Biotechnology continues to drive innovation in the production of pharmaceuticals, biofuels, and other valuable compounds, leveraging the power of microbial systems for enhanced yield and sustainability. Genome-scale metabolic (GSM) modeling has become an essential approach in this field that enables a guide for target genetic modifications and optimization of metabolic pathways for various industrial applications. While single-species GSM models have traditionally been employed to optimize strains like Escherichia coli and Lactococcus lactis, the integration of these models into community-based approaches is gaining momentum. Constraint-based metabolic community (CBMc) models, which simulate the interactions between multiple microbial species, offer a more comprehensive understanding of metabolic networks and their potential for improvement of biotechnological processes on pilot- and industrial scales. However, there are few platforms that effectively support both single-species GSM and CBMc models, limiting the accessibility and application of these advanced modeling techniques. In this study, we demonstrate the use of the BioUML platform, one of the few tools that provides simultaneous investigation of both single-species and community models. We harnessed the platform to reconstruct and analyze microbial interactions within a synthetic consortium of widely used biotechnology microorganisms: methanotroph, Methylococcus capsulatus Bath and engineered Escherichia coli strains under oxygen- and nitrogen- limited conditions. Our research highlights the potential of E. coli to reduce the accumulation of inhibitory by-products like acetate, thereby enhancing the metabolic efficiency and growth of M. capsulatus. The study underscores the utility of CBMc models in optimization of microbial consortia for biotechnological applications, offering new strategies for the sustainable production of biofuels, single-cell proteins, and other high-value compounds.

Candida albicans, a significant human pathogen due to its biofilm-forming ability, poses a serious health risk. This study investigates the antifungal activity and biofilm inhibition potential of Tea Tree Oil (TTO) from Melaleuca alternifolia cultivated in Vietnam. Renowned for its bioactive compounds, including terpinen-4-ol, 1,8-cineole, and terpinolene, the essential oil of M. alternifolia, commonly referred to as TTO, is scrutinized for its impact on C. albicans. The study meticulously determines the Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC) of TTO against C. albicans in both planktonic and biofilm states. Additionally, it assesses the biofilm-forming capacity of C. albicans and establishes the Minimum Biofilm Inhibitory Concentration (MBIC) and Minimum Biofilm Eradication Concentration (MBEC) of TTO. Employing a liquid culture dilution method within a defined concentration range of 0.1 μL/mL to 10 μL/mL, the experimental approach adheres to established protocols. The outcomes reveal a noteworthy MIC and MIC₈₀ of 0.1 μL/mL and 2 μL/mL, respectively, and an MFC of 10 μL/mL for TTO against C. albicans. Furthermore, TTO exhibits an MBIC of 2 μL/mL, showcasing its efficacy in inhibiting biofilm formation. However, the MBEC of TTO remains uncertain, possibly due to insufficient concentration tested, which may not effectively disrupt over 50% of the pre-formed biofilm mass within 24 hours. These findings highlight TTO’s significant potential as both an antifungal agent and a biofilm inhibitor against C. albicans, providing essential insights that could guide the development of new therapeutic approaches for treating fungal infections.

Tryptophan is an essential aromatic amino acid widely used in the pharmaceutical, agricultural, and food industries. Microbial fermentation, mainly using Escherichia coli, has become the preferred method for its production due to sustainability and lower costs. Optimizing tryptophan production requires careful control of various fermentation parameters, including nutrients, pH, temperature, and dissolved oxygen (DO) levels. Glucose, as the primary carbon source, must be fed at controlled rates to avoid metabolic overflow, which leads to byproduct accumulation and reduced production efficiency. Nitrogen sources, both organic (such as yeast extract) and inorganic (like ammonium), influence biomass growth and tryptophan yield, with ammonium levels requiring careful regulation to avoid toxic accumulation. Phosphate enhances growth but can lead to byproduct formation if used excessively. pH is another critical factor, with an optimal range between 6.5 and 7.2, where enzyme activity is maximized. Temperature control promotes growth and production, particularly between 33°C and 36°C. High DO levels increase tryptophan titers by boosting the pentose phosphate pathway and reducing byproducts like acetate. Furthermore, surfactants and supplements such as betaine monohydrate and citrate help alleviate osmotic stress and enhance precursor availability, improving production efficiency. Careful manipulation of these parameters allows for high-density cell cultures and significant tryptophan accumulation, making microbial fermentation competitive for large-scale production.