The gut, often referred to as the "second brain," has significant contributions in the maintaining of the good bacteria contribute enormous role such as digestion, produce essential vitamins, support the immune system, and protect against harmful bacteria. The beneficial flora including Akkermansia muciniphila, Adlercreutzia equolifasciens, Barnesiella, Christensenella minuta, and Oxalobacter formigenes, along with their derived bioactive metabolites emerged as a key player in maintaining host metabolic and immune health. Dietary choices such as blending of prebiotic, fermented, symbiotic, anti-inflammatory foods, and secondary metabolites from a wide variety of plants and fruits promotes the diversity, composition, and stability of beneficial intestinal microbes. The colourful plant foods rich in phytochemicals bioactive compounds such as carotenoids, flavonoids, polyphenols, alkaloids, anthocyanins, and capsaicin offer a wide array of unique properties such as analgesics, antioxidants, anti-inflammatory, antimicrobials effect and promoting the abundance of beneficial gut bacteria and their bioactive metabolites confer numerous health-promoting effects. Here, we present knowledge about most beneficial gut bacteria and their derived metabolites in terms of their sources and health benefits. Finally, we discuss best foods that skew towards promoting healthy intestinal microbes.

The present study investigated the photoprotective effect of the ultrasonic-assisted ethanol extract (USHE) from Sargassum horneri, a brown seaweed containing fucosterol (6.22 ± 0.06 mg/g) and sulfoquinovosyl glycerolipids (C23H43O11S, C25H45O11S, C25H47O11S, C27H49O11S), against oxidative damage in ultraviolet B (UVB)-exposed HaCaT keratinocytes. USHE indicated the antioxidant activity with increasing ferric-reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging. Moreover, USHE increased cell viability by markedly reducing the production of intracellular reactive oxygen species (ROS) in UVB-exposed HaCaT keratinocytes. Additionally, USHE reduced the apoptosis, and sub-G1 cell population, and increased the mitochondrial membrane potential. Moreover, USHE modulated the protein expression levels of anti-apoptotic molecules (Bcl-xL, Bcl-2, and PARP), and pro-apoptotic molecules (Bax, cleaved caspase-3, p53, cleaved PARP, and cytochrome C). This modulation accorded with the upregulation of cytosolic heme oxygenase (HO)-1, NAD(P)H quinone oxidoreductase 1 (NQO 1), and nuclear factor erythroid-2-related factor 2 (Nrf2), collectively known as components of the antioxidant system. These findings suggest that USHE has a photoprotective effect in UVB-exposed HaCaT keratinocytes and can be utilized to develop cosmeceuticals for UVB protection.

The serious impact of Alternaria alternata on jujube black spot disease has seriously affected the quality and yield of jujube, constraining the sustainable development of the jujube industry. The purpose of this study was to isolate and screen highly effective biocontrol strains of jujube black spot disease from jujube rhizosphere soil. Thirty-three soil samples were collected from 4 regions in southern Xinjiang. The strains with antagonistic effects were isolated and screened by the dilution spread method and plate confrontation method and identified by morphological, physiological, and biochemical characteristics, as well as 16S rDNA, gyrB, and rpoB gene sequences. Indoor and field efficacy experiments were conducted to determine their biocontrol effect. A total of 110 strains with antibacterial activity were selected, and one strain, Bacillus velezensis 26-8, with a stable antagonistic effect was further tested. Biological characteristic experiments showed that strain 26-8 could grow at NaCl concentrations of 0.5%-10% and pH 4.0-9.0. The biocontrol experiment results showed that Bacillus velezensis 26-8 could achieve an 89.83% control effect against black spot disease. In conclusion, strain 26-8 has good salt and alkali tolerance, exerts a good control effect on jujube black spot disease, and is worthy of further study.

Microfluidic systems offer controlled microenvironments for cell-to-cell and cell-to-stromainteractions, which have precise physiological, biochemical, and mechanical features. The opti-mization of their conditions to best resemble tumor microenvironments constitutes an experimentalmodeling challenge, particularly regarding carcinogenesis in the central nervous system (CNS), giventhe specific features of the blood–brain barrier (BBB). Gel-free 3D microfluidic cell culture systems(gel-free 3D-mFCCSs), including features such as self-production of extracellular matrices, providesignificant benefits, including promoting cell–cell communication, interaction, and cell polarity. Theproposed microfluidic system consisted of a gel-free culture device inoculated with human brainmicrovascular endothelial cells (HBEC5i), glioblastoma multiforme cells (U87MG), and astrocytes(ScienCell 1800). The gel-free 3D-mFCCS showed a diffusion coefficient of 4.06 × 10−9 m2·s−1, and itreconstructed several features and functional properties that occur at the BBB, such as the vasculo-genic ability of HBEC5i and the high duplication rate of U87MG. The optimized conditions of thegel-free 3D-mFCCS allowed for the determination of cellular proliferation, invasion, and migration,with evidence of both physical and biochemical cellular interactions, as well as the production ofpro-inflammatory cytokines. In conclusion, the proposed gel-free 3D-mFCCSs represent a versatileand suitable alternative to microfluidic systems, replicating several features that occur within tumormicroenvironments in the CNS. This research contributes to the characterization of microfluidicapproaches and could lead to a better understanding of tumor biology and the eventual developmentof personalized therapies. (PDF) A Versatile Microfluidic Device System that Lacks a Synthetic Extracellular Matrix Recapitulates the Blood-Brain Barrier and Dynamic Tumor Cell Interaction. Available from: https://www.researchgate.net/publication/384802118_A_Versatile_Microfluidic_Device_System_that_Lacks_a_Synthetic_Extracellular_Matrix_Recapitulates_the_Blood-Brain_Barrier_and_Dynamic_Tumor_Cell_Interaction#fullTextFileContent [accessed Oct 10 2024].

Background: Avaram Amrit, derived from a traditional medicinal plant, has shown potential in diabetes management. This study aimed to evaluate its clinical efficacy in glycemic control. Methods: A 12-week, open-label clinical trial was conducted with 9 type 2 diabetes patients. Participants received Avaram Amrit extract daily. Fasting and postprandial glucose levels were measured bi-weekly, with HbA1c assessed at baseline and week 12. Results: Fasting glucose levels decreased significantly from 252.54 ± 71.99 mg/dL at baseline to 195.50 ± 55.01 mg/dL at week 12 (p<0.001), representing a 22.6% reduction. Mean HbA1c decreased by 1.25 percentage points. Postprandial glucose levels showed consistent improvement across the study period. A strong positive correlation was observed between initial HbA1c and fasting glucose (r=0.89). Conclusion: Avaram Amrit demonstrated significant improvement in glycemic control over 12 weeks. The consistent decrease in both short-term (glucose levels) and long-term (HbA1c) measures indicates a positive response to the intervention. While promising, larger randomized controlled trials are necessary to confirm these findings and establish Avaram Amrit's role in diabetes management.

One of the most common nutritional problems in athletes is iron deficiency. Although a lot of research has been done, the problem is still not solved. It is believed that iron deficiency is basically a lack of iron intake due to low dietary intake. Therefore, athletes who are found to be iron deficient are required to increase their iron intake (supplementation or intravenous administration). However, there are many supplements available on the market today, and if the problem is simply a lack of iron intake, it is a nutritional problem that should have been solved. In other words, there are reasons why the problem cannot be solved by simply taking iron, and it is important to conduct further research on the relationship between iron status and physical activity in the future. 　In this paper, we summarize our recent research, focusing on the studies we have done so far.

The drug discovery process is a complex and time-consuming endeavor that can greatly benefit from advancements in computational biology. This study explores the implementation of GPU-accelerated computational methods to enhance molecular docking simulations and virtual screening techniques, two crucial components of drug discovery. By leveraging the parallel processing capabilities of Graphics Processing Units (GPUs), we significantly accelerate the analysis of protein-ligand interactions and the identification of potential lead compounds. Our approach utilizes optimized algorithms and software frameworks to achieve substantial speedups over traditional CPU-based methods. Results demonstrate that GPU-accelerated molecular docking simulations and virtual screening can reduce computation times by up to 10-fold, enabling rapid and accurate identification of promising drug candidates. This accelerated workflow has the potential to revolutionize the drug discovery pipeline, facilitating the development of novel therapeutics and reducing the time and cost associated with bringing new treatments to market.

Chronic wounds have a significant impact on patients' quality of life, necessitating the management of pain, infection, bleeding, and emotional challenges. Debridement, which involves the removal of nonviable tissue, is crucial for promoting wound healing. In addition to surgical methods, cost-effective alternatives such as local solutions and ointments with biological properties have been explored. The use of natural compounds with anti-inflammatory, antibacterial, and collagen-synthesizing abilities holds promise for wound healing. This study aimed to assess the safety and effectiveness of a wound-healing ointment containing bioactive ingredients derived from medicinal plants (extracts, essential oils, and vegetable oils). Pre-clinical tests were conducted following standardized protocols. The chemical composition of the ointment was characterized using Fourier transform infrared (FTIR) spectroscopy to gain insights into its synergistic action. FTIR analysis revealed similarities between the product's spectrum and that of bioactive compounds. The in vitro tests demonstrated that all formulations of the ointment induced no cell death, DNA damage, or acute toxicity in cell cultures. No lethal dose was observed, indicating the safety of the ointment at all concentrations. In vivo preclinical analyses demonstrated no adverse responses being effective in the healing process, similar to the gold standard (Silver Sulfadiazine). The regenerative ointment displayed excellent biocompatibility and bioactivity in vitro and in vivo studies, contributing to the development of innovative and sustainable wound management therapies.

Plasmonic-based biosensors have gained prominence as potent optical biosensing platforms in both scientific and medical research, attributable to their enhanced sensitivity and precision in detecting biomolecular and chemical interactions. However, the detection of low molecular weight analytes with high sensitivity and specificity remains a complex and unresolved issue, posing significant limitations for the advancement of clinical diagnostic tools and medical device technologies. Notably, abnormal cholesterol levels are a well-established indicator of various pathological conditions; yet, the quantitative detection of its free form is complicated by its small molecular size, pronounced hydrophobicity, and the necessity for mediator molecules to achieve efficient sensing. In the present study, a novel strategy for cholesterol quantification was developed, leveraging a plasmonic optical readout in conjunction with a highly specific cholesterol-binding peptide (C-pept) as a biorecognition element, anchored on a functionalized silica substrate. The resulting biosensor exhibited an exceptionally low detection limit of 21.95 µM and demonstrated a linear response in the 10-200 µM range. This peptide-integrated plasmonic sensor introduces a novel one-step competitive method for cholesterol quantification, positioning itself as a highly sensitive biosensing modality for implementation within the AVAC platform, which operates using reflective dark-field microscopy.

Diagnostics often require specialized equipment and trained personnel in laboratory settings, creating a growing need for point-of-care tests (POCT). Among genetic testing methods, Loop-mediated Isothermal Amplification (LAMP) offers a viable solution for developing genetic POCT due to its compatibility with simplified devices. This study aimed to create a genetic test that integrates all steps from sample processing to results while minimizing the complexity, handling, equipment, and time required. Several challenges were addressed to achieve this goal: 1) Developing a buffer for bacterial DNA extraction that is compatible with both LAMP and immunochromatographic tests; 2) Adapting the LAMP protocol for its use with the device; and 3) Optimizing the detection protocol for specific test conditions, with a lateral flow immunoassay format selected for its POCT compatibility. Following these developments, the test was validated using Escherichia coli and non-E. coli strains. A portable heating station was also developed to enable amplification without costly equipment. The resulting genetic POCT achieved 100% sensitivity and 85% specificity, with results available in 60 to 75 minutes. This study demonstrated that our closed-method POCT efficiently performs DNA extraction, amplification, and detection for bacterial identification. The test’s simplicity and cost-effectiveness will support its implementation in various settings.

Microorganisms play a pivotal role in ecology, medicine, agriculture, and biotechnology. To comprehend their diversity, physiology, and functions, isolation and characterization are imperative. This review provides a concise overview of prevalent molecular biotechniques for microbial isolation. It describes the advantages, constraints, and practical applications of both culture-dependent and culture-independent approaches. Discussed methods encompass selective/differential media, 16S rRNA gene sequencing, metagenomics, fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR)-based techniques, and single-cell genomics. Emerging high-throughput sequencing and single-cell isolation methods exemplify cutting-edge procedures explored herein. Integration of these molecular tools offers comprehensive insights into microbial-diversity, community structure, metabolic capabilities, and functional interplays.

This perspective describes the current status and future prospects of developing long- to ultralong-acting anti-obesity peptides. First, we review the current status of lipidation, PEGylation, and Fc fusion technologies to obtain long-acting peptides administered once weekly. Next, we describe the approach and current results of using macromolecular peptide prodrugs with pre-programmed releasable linkers to achieve longer-acting peptides that can be administered weekly or monthly. Finally, we posit novel modifications of the latter technology that could provide ultralong half-lives of over one month by advantageously exploiting the clearance rates of released peptides. As examples, we posit that prodrugs of low-clearance GLP-1 agonists derived from peptides already modified by lipidation, PEGylation, and Fc fusion could produce ultralong-acting agonists with dosing intervals reaching 3 to even 6 months.

Since the discovery that Histone deacetylase inhibitors (HDCAi) could enhance radiation response, a number of HDACi, mainly pan-HDAC inhibitors, have been studied either as monotherapy or in combination with photon irradiation or chemotherapeutic drugs in the management of breast cancer. Studies on combination of HDACi and particle type of radiation remain limited. CUDC-101 is a multitarget inhibitor of Histone deacetylases (HDACs), epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2(HER-2). In this paper, the effectiveness of CUDC-101 in enhancing radiation response to protons irradiation was studied in MCF-7, MDA-MB-231 and MCF-10A cell lines, using X-rays as reference radiation. γ-H2AX foci assays showed increased sensitivity to CUDC-101 in MDA-MB-231 cell line compared to the MCF-7 cell line. In both cell lines, induction of apoptosis was enhanced in CUDC-101 pre-treated cells compared to radiation (protons or X-rays) alone. Increased apoptosis was also noted in CUDC-101 pre-treated cells in the MCF-10A cell line. Cell cycle analysis showed increased G2/M arrest by CUDC-101 mono-treatment as well as combination of CUDC-101 and X-irradiation in the MDA-MB-231 cell line. Collectively, the findings indicate that CUDC-101 effectively enhances response to both X-radiation and proton irradiation, in the triple negative MDA-MB-231 cell line. The study highlights that CUDC-101 holds potential in the management of triple negative breast cancer as monotherapy or in combination with X-irradiation.

Jararhagin-C (JarC) is a protein from the venom of Bothrops jararaca consisting of disintegrin-like and cysteine-rich domains. JarC shows a modulating effect on angiogenesis and remodeling of extracellular matrix constituents improving wound healing in mice experimental model. JarC is purified from crude venom and the yield is less than 1 %. The aim of this work was to obtain the recombinant form of JarC and to test its biological activity. For this purpose, the bicistronic vector pSUMOUlp1 was used. This vector allowed the expression of the recombinant toxin JarC (rJarC) in fusion with the small ubiquitin-related modifier (SUMO) as well as the SUMO protease Ulp1. After expression, this protease was able to efficiently remove SUMO from rJarC inside the bacteria. rJarC free from SUMO was purified at the expected molecular mass, and recognized by polyclonal anti-Jararhagin antibodies. In terms of biological activity, both the native and recombinant forms showed no toxicity to the HUVEC cell line CRL1730 and were effective in modulating cell migration activity in experimental in vitro model. These results demonstrate the successful production of rJarC and the preservation of its biological activity, which may facilitate further investigations into the therapeutic potential of this snake venom-derived protein.

The salivary microbiota of Italian and sub-Saharan African individuals was investigated using Nanopore Sequencing Technology (ONT- Oxford Nanopore Technologies). We detected variations in community composition in relation to endogenous (ethnicity, sex and diplotypic variants of the TRPV1 gene) and exogenous (sensitivity to capsaicin) factors. The results showed that, Prevotella, Haemophilus, Neisseria, Streptococcus, Veillonella and Rothia, are the most abundant genera, in accordance with the literature. However, alpha diversity and frequency spectra differed significantly between DNA pools. The microbiota in African, male, TRPV1 bb/ab diplotype and capsaicin low sensitive DNA pools was more diverse than Italian, female, TRPV1 aa diplotype and capsaicin high sensitive DNA pools. Relative abundance differed at phylum, genus and species level.

Background/Objectives: The increasing incidence and mortality rates of liver cancer have heightened the demand for the development of effective anticancer drugs with minimal side effects. In this study, the roles of exosomes derived from liver cancer stem cells (LCSCs) with PRELI (Protein of Relevant Evolutionary and Lymphoid Interest) modulation, and their miRNAs were investigated to explore the therapeutic for liver cancer. The purpose of this study is to identify a more effective and less toxic anticancer agent beyond the well-known Notch, p51, and Hippo signaling pathways, and to specifically propose the potential of PRELI-induced exosomes as an anticancer therapeutic candidate. Methods: Various techniques, such as miRNA profiling, flow cytometry, western blotting, and immunocytochemistry, were used to evaluate the effects of ex-osomes under PRELI up- and downregulation. Results: PRELI upregulation altered expression levels of miRNAs, including hsa-miR-378a-3p (involved in stem-like properties), hsa-miR-25-3p (contributing to cell proliferation), and hsa-miR-423-3p (driving invasiveness). Exosomes with downregulated PRELI inhibited AKT/mTORC1 signaling, while this signaling pathway was ac-tivated in LCSCs transfected with the candidate miRNAs. Under PRELI up-regulation, exosomes showed increased surface marker expression, promoting cancer progression. Conclusions: The modulation of PRELI in LCSCs affects miRNA expression significantly, revealing candidate miRNA targets for liver cancer treatment. Exosomes with PRELI down-regulation show potential as a novel therapeutic strategy.

Objective: To analyze and investigate the impact of early rehabilitation training based on developmental sequencing on the growth and development of late preterm infants. Methods: This study focused on preterm infants admitted to our hospital from June 2021 to November 2021. Strict screening was applied to select 54 cases for inclusion. Clinical data were collected for all study subjects, and based on different intervention plans, they were divided into a control group and an experimental group, with 27 cases in each group. The experimental group received intervention based on a developmental sequencing early rehabilitation model in addition to routine care, while the control group only received routine care intervention. The growth and development of the two groups of preterm infants were compared during the study, including physical indicators, intellectual-motor skills, neurobehavioral assessments, and developmental monitoring. Results: The clinical data of the two groups of preterm infants were comparable, with no significant differences (P>0.05). The experimental group exhibited significantly higher values in physical indicators such as weight, head circumference, and height compared to the control group (P<0.05). Intellectual-motor scores (MDI and PDI) of the experimental group were higher than those of the control group (P<0.05). The NBNA scale scores for various dimensions were higher in the experimental group compared to the control group (P<0.05). The DQ values of the experimental group were higher than those of the control group (P<0.05). Conclusion: Early rehabilitation training based on developmental sequencing has a positive impact on the growth and development of late preterm infants. Through systematic and sequenced rehabilitation interventions, various aspects of development in late preterm infants can be promoted effectively. This approach not only improves physical development but also enhances intellectual-motor skills, neurobehavioral development, and overall developmental levels. The results are significant, indicating the worthiness of widespread application and promotion.

Plant-associated yeasts can also be part of soils’ and pollinating-insects’ microbiomes, where they play significant ecological roles. Indeed, in previous studies, we have shown that yeasts may help pollinators find nectar, which is crucial for their nutrition and for the reproduction of many angiosperms, and, in the soil, can act as plant-growth promoters. Given the importance of yeasts for plant development, in this review, we first delve deep into the biochemical processes behind the ecological role of these microorganisms in soil, insects, and in direct association with plants. Based on this premise, we discuss the influence of this relationship on agricultural production and the possible effects of pesticide use on the yeast microbiota. Finally, we address the most relevant studies in the literature that support the potential of these microorganisms (either indigenous or genetically engineered) as bioremediation agents for soils and foods contaminated by pesticides. Our review indicates that yeasts can be satisfactorily employed in organic agriculture to increase plant growth or bioremediate contaminated soil or food.

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.

Research in seaweed cultivation technologies aims to increase production and reduce costs, leading to more efficient and sustainable processes. In this study, we analyzed the outdoor production of Ulva compressa cultured in summertime at different stocking densities of 0.6, 0.8 and 1.0 kg Fresh weight (FW) m-2 in a raceway photobioreactor with 30 m2 surface (3000L) and its relation to pho-tosynthetic activity. Under the experimental conditions of high temperature (> 28-30 oC) and pH > 9 in culture water, higher seaweed density resulted in lower specific growth rate. The biomass pro-duction has been related to photosynthetic activity by using in vivo chlorophyll a fluorescence. Dynamic photoinhibition was observed at noon, which was less severe in cultures with higher algal densities. However, photosynthesis recovered in the afternoon. Seaweeds that were acclimatized for a week to the conditions of 1.0 kg FW m-2 stocking density showed an increase in biomass growth and absence of photoinhibition compared to non-acclimatized thalli. In conclusion, by using high densities (1.0 kg FW m-2), U. compressa presented the highest growth rate in a mid-scale raceway photobioreactor under stressed conditions related to the increase of photosynthetic capacity.

Interferons (IFNs) are cytokines involved in the immune response with synergistic regulatory action. They are therapeutics for various viral and proliferative conditions, with proven safety and efficacy. Their clinical application presents difficulties due to the molecules' size, degradation, and pharmacokinetics. We developed a controlled release system for viral respiratory tract infections. A core-shell nanoparticle was prototyped, which hydrolyzed the shell (polyvinylpyrrolidone), releasing the active ingredients IFN-α and IFN-γ. The core (chitosan) degraded slowly, with a controlled release of IFN-α. The primary and rapid effect of the combination of interferons ensured an antiviral and immunoregulatory response from day one, induced by IFN-α and enhanced by IFN-γ. The multilayer design demonstrated an optimal toxicity profile. This formulation is an inhaled dry powder targeted for the non-invasive intranasal route. This prototype would ensure greater bioavailability, controlled release, fewer adverse effects, and robust biological action thanks to the simultaneous impact of both molecules.

The use of probiotics has been gaining popularity in terms of inclusion into human diets over recent years. Based on properties exerted by these living organisms, several benefits have been elucidated and conferred to the host. Bacteria has been more commonly used in probiotic preparations, in comparison to yeast candidates, however, yeast exhibit several beneficial properties such as the prevention and treatment of diarrhoea, production of antimicrobial agents, prevention of pathogen adherence to intestinal sites, maintain microbial balance, modulation of the immune system, resistant to some antibiotics, amongst others. This review details the use of yeast organisms as biotherapeutics and has a special focus on production considerations and their formulation into different delivery formats.

This study used a completely randomized design with a bifactorial arrangement in triplicate to evaluate dif-ferent concentrations of flavonoid extracts from orange (Citrus sinensis), onion (Allium cepa), tamarillo (So-lanum betaceum) and cocoa (Theobroma cacao) peels in combination with the yeast Saccharomyces cerevisiae to inhibit the growth of Listeria monocytogenes in fruit juices. The shells were dehydrated at 50-57 °C for 8 hours and pulverized. The dehydration yields were: orange (32 %), onion (13 %), tamarillo (14 %) and cocoa (34 %). Flavonoids were extracted with Soxhlet using 70% ethanol for 5 hours, concentrating with a rotary evaporator. They were quantified by UV spectrophotometry and the Folin-Ciocalteu method, being the highest in orange (300,75-372,60 mg Q/L) and onion (205,70-534,96 mg Q/L), while tamarillo presented the lowest, with possible errors in the measurement. Soluble solids losses ranged from 2,30 g (cocoa) to 5,27 g (orange). Cocoa and orange extracts showed greater potential as natural preservatives, inhibiting the growth of Listeria monocytogenes in fruit juices, with a minimum inhibitory concentration of 300 µg/mL and 400 µg/mL, re-spectively.

No data on wild mammals run over along the roads in Osijek-Baranja County, in the north-eastern part of Croatia, have been available so far. Mammals run over from 2016 to 2024 on the Zmajevac-Osijek road, 33.4-km long, were recorded using the road sampling method while travelling back and forth from Zmajevac to Osijek. A total of 86 run-over specimens of wild mammals, classified into eight species and one subspecies, were recorded. The northern white-breasted hedgehog, Erinaceus roumanicus, topped the list with 38 individuals, followed by the European badger, Meles meles, with 23 individuals. Together, these two species amounted to 70.93% of the run-over mammal fauna. Other species and subspecies were recorded in lower numbers. The number of wild mammals run over per kilometer of the observed road section was on the rise from 0.09 to 0.20 in the period from 2016 to 2022, while in 2023 it increased to 0.77 and in 2024 to 0.86. The high proportion of run over wildlife per kilometer is likely due to the high volume of traffic, which disrupts wildlife activities during both day and night.

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.

Retinal diseases are an important focus of in vivo gene therapy, for which adeno-associated virus is a preferred vector. Intravitreal injection is a highly desirable delivery route because it is much simpler, safer and inexpensive than the main alternative subretinal injection. However, existing capsids have low efficiency, require high doses and are susceptible to neutralization by antibodies that are present in the vitreous. Using AAV5 as starting material, we applied a combination of directed evolution and rational design to develop PT1, a novel AAV capsid capable of efficiently transducing the retina after intravitreal injection. The most striking property of PT1 is its remarkable ability to evade preexisting neutralizing antibodies, thanks to its multiple mutations spread over large areas of the capsid surface, disrupting several epitopes. Our results suggest that PT1 has a strong potential as a useful candidate capsid for human retinal gene therapy.

Bacterial structures formed from the outer membrane and the periplasm components carry biomolecules to expel cellular material and interact with other cells. These outer membrane vesicles (OMVs) can encapsulate bioactive content, which gives OMVs a high potential as alternative drug delivery vehicles or as a platform for novel vaccine development. Single-gene mutants of the hypervesiculating strain Escherichia coli JC8031 were engineered to further enhance OMV production based on metabolic network modelling and in silico gene knockout design (ΔpoxB, ΔsgbE, ΔgmhA, and ΔallD). Mutants were experimentally obtained by genome editing using CRISPR-Cas9 and tested for OMVs recovery observing an enhanced OMV production in all of them. Lipidomic analysis through LC-ESI-QTOF-MS was performed for OMVs obtained from each engineered strain and compared to the wild type E. coli JC8031 strain. The lipid profile of OMVs from the wild type E. coli JC8031 did not change significantly when compared to the mutant strains. The obtained results suggest that the vesicle production can be further improved while the obtained vesicles are not altered in their composition, allowing further study for stability and integrity for use in therapeutic settings.

This study evaluated the post-thaw viability and in vitro fertility of ejaculated and epididymal semen from Pantaneiro bulls and characterized cell-free DNA (cfDNA) in fresh seminal plasma. Semen from five bulls was collected via electroejaculation or post-mortem epididymal extraction. Fresh semen parameters and cfDNA concentrations were assessed before cryopreservation. Post-thaw sperm kinetics were evaluated using CASA at 0 and 6 hours of incubation, and in vitro embryo development was analyzed following IVF. Data were assessed using ANOVA and logistic regression. Ejaculate samples exhibited more morphological defects than epididymal samples (15.8% vs. 1.8%, P≤0.05). Post-thaw, epididymal semen showed higher total (87.2% vs. 32.4%) and progressive (67.1% vs. 14.4%) motility at 0 h (P≤0.05), and higher motility at 6 h (38.9% vs. 11.0%, P≤0.05). In vitro fertility did not differ significantly between ejaculated (n=525 oocytes) and epididymal (n=500 oocytes) semen groups in terms of cleavage (49.6% vs. 44.2%) and blastocyst formation on D7 (26.1% vs. 22.2%, P&gt;0.05). cfDNA concentration in fresh semen ranged from 11.4 to 50.9 ng/µL. These findings indicate that epididymal sperm from Pantaneiro bulls retain high post-thaw viability and fertility. Additionally, cfDNA characterization in seminal plasma contributes to indigenous cattle preservation and advances in male fertility research.

Background/Objectives: Listeria monocytogenes is a foodborne pathogen that can infect humans and animals, causing non-invasive gastrointestinal listeriosis or invasive listeriosis. The objectives of this study were to determine: the genetic diversity of L. monocytogenes; the genes associated with the resistance to antibiotics, benzalkonium chloride (BC), and CdCl₂, and biofilm formation. Methods: Seventeen strains of L. monocytogenes isolated fresh fruits and vegetables were selected for this study. Genetic diversity and the genes associated with antibiotic resistance were determined by PCR amplification. Susceptibility to antibiotics were determined using the agar diffusion method. Resistance to BC and CdCl₂ was determined using the minimum inhibitory concentration method. Capacity for biofilm formation was evaluated using the crystal violet staining method. Results: This study demonstrates that the isolates of L. monocytogenes belong to serotypes 1/2a (29.4%) and 1/2b (70.5%). Potential hypervirulent strains containing 70.5% Listeria pathogenicity island 1 (LIPI-1-2), 17.6% (LIPI-1-3), and 11.7% (LIPI-1-4) were also identified. Antibiotic susceptibility tests show that L. monocytogenes exhibit six different multiresistant patterns with a MAR index of ³ 0.46 (70.5%); besides, the gen Ide, tetM, and msrA were detected in 52.9%, 29.4%, and 17.6%, respectively, associated with efflux pump Lde, tetracycline and ciprofloxacin resistance. Phenotypic test shows that 58.8% cadmium-resistant L. monocytogenes have a co-resistance to BC of 23.5%. Subsequently, all strains of L. monocytogenes exhibited moderate biofilm. Conclusions: This study contributes to the persistence and genetic diversity of strains L. monocytogenes isolated fresh fruits and vegetables; in addition, resistance to CdCl₂ and correlation with co-resistance to BC disinfectant used in the food industry.

Genetic engineering has revolutionized our ability to modify microorganisms for various applications in agriculture, medicine, and industry. This review examines recent advances in genetic engineering techniques for bacteria, fungi, and oomycetes, with a focus on CRISPR-Cas systems. In bacteria, CRISPR-Cas9 has enabled precise genome editing, enhancing applications in antibiotic production and metabolic engineering. For fungi, despite challenges associated with their complex cell structures, CRISPR/Cas9 has advanced the production of enzymes and secondary metabolites. In oomycetes, significant plant pathogens, modified Agrobacterium-mediated transformation and CRISPR/Cas12a have contributed to developing disease-resistant crops. The review provides a comparative analysis of genetic engineering efficiencies across these microorganisms and addresses ethical and regulatory considerations. Future research directions include refining genetic tools to improve efficiency and expand applicability in non-model organisms. This comprehensive overview highlights the transformative potential of genetic engineering in microbiology and its implications for addressing global challenges in agriculture, medicine, and biotechnology.

Chronic wounds remain a major challenge in healthcare, with conventional dressings often leading to infections and lacking advanced healing capabilities. This study evaluated the wound healing potential of NIMO-CH, a cellulose hydrogel derived from nipa fronds and enriched with indigenous microorganisms, particularly Lacticaseibacillus paracasei BCRC-16100. Using ICR mice with excisional wounds, three treatments were compared: no treatment, NIMO-CH, and DuoDERM. Results showed that NIMO-CH and DuoDERM both achieved 100% wound closure, with NIMO-CH-treated wounds showing complete healing with hair growth by day 18-20, while untreated wounds healed by day 20. Minimal scarring was observed in both NIMO-CH and DuoDERM groups. Histological analysis confirmed similar healing processes, including granulation tissue formation and moderate inflammatory responses, with no significant differences in collagen fiber orientation. Furthermore, genome sequencing revealed that Lacticaseibacillus paracasei BCRC-16100 contained genes sodA and gsiC, responsible for coding enzymes necessary for the wound healing process. These results suggest that NIMO-CH could be an effective alternative for wound care applications.

Organisms from the genus Streptomyces feature actinobacteria with complex developmental cycle and great ability to produce a variety of natural products, which is possible due to complicated crosstalk between primary and secondary metabolism. These soil bacteria produce more than 2/3 of antibiotics used in medicine, and a large variety of bioactive compounds for industry and agricultural use. Although Streptomyces spp. have been studied for decades, the engineering of these bacteria remains challenging, and available genetic tools rather limited. Recent advancements in genetic manipulation of Streptomyces involving proposal of CRISPR/Cas9-based workflows as well as synthetic components (e.g. promoters, ribosome-binding sites, terminators, reporter genes) allowed to facilitate the turnaround time of strain engineering, but still has strain-specific limitations. However, a new perspective offered by synthetic biology to exploit the potential of existing and novel pathways in primary and secondary metabolism allows combining of different biosynthetic steps originating from diverse bacteria using a limited toolbox. Manipulation of interplay between primary and secondary metabolism proposes strategies to overcome strain-specific difficulties in engineering, leveraging insights in Streptomyces-specific physiological features. In this review, developments of these approaches for Streptomyces engineering are discussed and an overview of the synthetic biology developments is provided.

Indoor air quality (IAQ) is a critical determinant of human health, with fungal contamination posing significant risks. In 2023, a total of 160 samples were collected from 40 selected rooms. These samples were cultured on Sabouraud Dextrose Agar (SDA) medium using a portable air sampler and incubated at 28°C for 7 days. Initial identification of fungal isolates was performed based on their morphology. Suspected isolates underwent further analysis using internal transcribed spacer (ITS) rDNA analysis. The results revealed 11 species across 8 genera in various rooms. The most prevalent fungi were Aspergillus niger (183 CFU) , Penicillium chrysogenum (139 CFU) and Cladosporium halotolerans (135 CFU). On the other hand, Curvularia hawaiiensis and A. ustus were rarely isolated (10 and 19 respectively). Effective treatment involves a combination of preventive measures, such as controlling humidity and ensuring proper ventilation, alongside targeted interventions like antifungal treatments and thorough cleaning. Continuous monitoring and management of indoor environments are essential to reduce fungal contamination, enhance air quality, and protect human health from the harmful effects of airborne fungi. This abstract underscore the importance of addressing indoor fungal contamination to promote safer, healthier living and working spaces.

Adeno-associated viruses (AAVs) have been used for liver gene therapy. Hemgenix and Roctavian are AAV-based treatments for hemophilia B and A, respectively. They cost $3 million and $2.8 million per dose, respectively. While AAV vectors may eventually be cheaper to mass produce, a bacteria-based DNA delivery system might be much cheaper for patients. Also, “bactofection” would allow for the delivery of much larger DNA packages. Such a bacterial system may now be possible, and a prototype for the liver could possibly be developed immediately.

The genetic transformation of Coffea arabica L. is an alternative strategy for obtaining plants with agronomic traits of interest that is less time-consuming than conventional breeding methods. Given the importance of coffee cultivation in Colombia, this study evaluated the main factors interfering with the genetic transformation of C. arabica using Agrobacterium tumefaciens. An effi-cient and reproducible method was accordingly established that involved the propagation of “early” embryogenic tissue in a liquid proliferation medium, supplemented with 3 mg l−1 BAP for eight months, which was subsequently sonicated for 300 sec in a suspension of LBA4405 OD600 of 0.5 harboring pC1301 and incubated in this same suspension for 1 hr. This plasmid contained the uidA gene under control of the 35S promoter. The Agrobacterium suspen-sion was subsequently removed from the embryogenic tissue using a micropipette, after which the tissue was deposited on filter paper to remove the remaining Agrobacterium suspension. The embryogenic tissue was co-cultured for four days in a solid differentiation medium supple-mented with 100 µM acetosyringone on filter paper. Subsequently, the tissue was post-cultured for four days in liquid differentiation medium under orbital shaking at 100 rpm with 300 mg l−1 Claforan (Hoechst), followed by selection with 50 mg l-1 hygromycin at 26 °C in the dark, with subcultures at 20-day intervals until somatic embryos were formed for subsequent culturing in germination medium. Molecular analysis confirmed the presence of the uidA gene in coffee seedlings transformed with strains LBA4405 and EHA105 and vectors pC1301 and pC2301. This method successfully enables the stable integration of genes of interest in the coffee plant ge-nome

The rat osteosarcoma cell line UMR-106 is widely used for the study of bone cancer biology. We have characterized UMR-106 with a combination of optical genome mapping (OGM), long-read sequencing (for identification of variants and 5mC methylation), and short-read RNA sequencing for expression. Genome sequence was compared to a Sprague-Dawley control animal, the strain from which UMR-106 was derived, and expression data were compared to a public rat osteoblast dataset. Using the COSMIC database to identify the most affected genes in human osteosarcomas (OS) we found somatic mutations in Tp53 and H3f3a in UMR-106. OGM identified a relatively small number of differences between the cell line and a strain-matched control animal but did detect a ~45 Mb block of amplification that included Myc on chromosome 7 which was confirmed by long-read sequencing. The amplified region showed several blocks of non-contiguous rearranged sequence implying complex rearrangements during their formation and included 14 genes reported as biomarkers in human osteosarcoma, many of which also showed increased transcription. A comparison of 5mC methylation from the nanopore reads of tumor and control identified several genes with distinct differences including the OS marker Cdkn2a. This dataset illustrates the value of long DNA methods for the characterization of cell lines and how inter-species analysis can inform about the genetic nature underlying mutations that underpin specific tumor types. The data should be a valuable resource for investigators studying osteosarcoma, in general, and specifically the UMR-106 model.

Tussilago farfara L. is a traditional medicinal plant valued for its potentially health-promoting metabolites. Its herbal raw material has been recognized and used since ancient times and continues to be widely used in traditional medicine. Introducing these plants to in vitro cultivation is a challenging task, but once the protocol is developed, such cultures can provide an abundant and inexhaustible source of plant material. In this study, we report the successful induction and growth of vigorous T. farfara L. callus in vitro. Callus induction was achieved on solid media in darkness, whereas it appeared inefficient under light conditions and in suspension culture. We present a detailed description of callus growth kinetics, morphological analysis, photosynthetic activity and biochemical parameters (including protein content and photosynthetic pigments) supported by histological studies. Furthermore, we observed potential for organogenesis and somatic embryogenesis. This method for in vitro propagation of T. farfara, along with callus culture maintenance, offers a wide range of applications in pharmacy and could benefit the environment by minimizing the depletion of natural populations of this species.

Gene therapy is a medical approach capable of correcting genetic and epigenetic alterations in pathological conditions, including cancer. Different strategies have been pursued for cancer gene therapy, aiming to restore the function of tumor suppressor genes or inhibit oncogenes. These comprise the use of CRISPR/Cas9 gene editing technology and therapeutic oligonucleotides, such as microRNAs, anti-microRNAs, small interfering RNAs, DNAzymes, and epigenetic modifiers. However, despite great potential, their broad clinical use is hindered due to the lack of safe and efficient systems for targeted delivery to cancer cells. To this purpose, aptamers could represent an ideal tool. They are short single-stranded oligonucleotides that bind receptors selectively present or overexpressed on cancer cell membranes, often undergoing cell-specific internalization. Thanks to this last feature, numerous internalizing aptamers have been investigated as targeting moiety to deliver therapeutic oligonucleotides, viral vectors, and nanoparticle-based systems in preclinical studies. Here, we summarize the state-of-the-art of aptamer applications in cancer gene therapy, providing key examples and discussing their advantages, limitations, and clinical perspectives.

Biofilm formation in unicellular eukaryotes, such as Candida albicans and Saccharomyces cerevisiae, represents a significant challenge in various sectors, including healthcare, agriculture, and industry. These biofilms are resistant to conventional antimicrobial treatments due to their protective extracellular matrix and complex regulatory mechanisms, such as quorum sensing (QS) and cyclic AMP (cAMP) signaling. Recent advances in the understanding of biofilm formation pathways have led to the identification of novel compounds that target these regulatory mechanisms. This review highlights breakthroughs from 2024 research, focusing on the inhibition of quorum sensing, disruption of extracellular matrix synthesis, and transcriptional/post-transcriptional regulation of biofilm-related genes. The application of these discoveries in Vietnam is particularly promising, given the country's challenges with biofilm-related infections in healthcare and productivity losses in agriculture and aquaculture. Natural product-based approaches, especially those derived from Vietnam's marine biodiversity, offer environmentally sustainable solutions for biofilm control. However, translating these laboratory findings into scalable, cost-effective applications faces challenges such as resistance development, environmental sustainability, and regulatory constraints. This review provides a comprehensive overview of compounds targeting biofilm regulatory mechanisms, evaluates their potential applications in Vietnam, and discusses future research directions, emphasizing the need for interdisciplinary collaboration to address the complexities of biofilm control.

Côte d’Ivoire has the main cashew tree culture area worldwide, but the cashew apple produced is still underutilized although its potential for industry. In the present study, the aim was to isolate and identify acetic acid bacterial strains, and to assess their potential use for cashew apple-based vinegar production. Vinegar mother from fermented juice was used to isolate acetic acid bacteria on a standard glucose-based medium. Physiological and biochemical tests followed by 16S rRNA gene analysis and phylogeny were used for isolate characterization. Moreover, their acetic acid production capacity was assessed. As results, five (05) strains of the Acetobacter genus were isolated. Phenotypic and phylogenetic analysis revealed that four of them, namely OYA2, OYA6, OYA9, and OYA10, belong to A. tropicalis / A. senegalensis species complex with 99,7% or 100% similarity. The last strain, OYA7, match to A. syzygii (99,7%). All the isolates were resistant to 15° alcohol (v/v) and grew well between pH 5.0 and 6.5. Their optimal growth temperatures varied between 27°C and 37°C, and only isolate OYA6 grows up to 40°C. They produced vinegar with yield (Yp/s , g/g) varying from 0.82 to 0.92, and acetic acid content (g/L) of 80.67 ± 2.1, 70.26 ± 0.9, 70.11 ± 1.7, 68.70 ± 1.5 and 67.22 ± 0.40 were obtained with OYA6, OYA7, OYA2, OYA10, and OYA9, respectively. Thus, the isolate OYA6 appeared as the best candidate for vinegar production, owing to its superior yield and thermotolerance ability that need to be further explored for industry use purpose.

Quorum sensing (QS) is a sophisticated bacterial cell-to-cell communication mechanism that allows bacteria to sense population density through the secretion and detection of diffusible small molecular signals. This process leads to the coordinated expression of specific genes at the transcriptional level. Over time, continuous research has elucidated the genetic elements and regulatory principles of QS. Recently, synthetic biology has leveraged these insights to construct genetic circuits incorporating QS components, enabling both intra-species and inter-species artificial communication. These QS-based genetic circuits hold significant potential for applications in biotechnology and biomedicine. This paper reviews several well-characterized microbial QS systems and their functional roles, while also introducing the application of QS-based genetic circuits in cellular communication across species. The paper further discusses the role of QS in the development of biological computing tools, population density regulation, and metabolic flux control, offering a forward-looking perspective on future advancements. For intra-species communication, the focus is on the use of QS systems in constructing biological computing tools, including toggle switches, biosensors, and logic gates in synthetic biology. These tools, designed on the QS mechanism, can more precisely coordinate cellular behavior by integrating biological control circuits to achieve spatial, temporal, and population-level regulation. In the context of inter-species communication, the introduction of QS systems plays a pivotal role in population density control and metabolic flow regulation. By recombining metabolic networks, QS enables the redistribution of metabolic flux in desired pathways, facilitating the regulation of population density and the co-culture of mixed strains. Moreover, combining QS with oscillator models has shown great potential in synchronizing microbial communities. In summary, in-depth research into QS mechanisms and their applications not only lays a solid foundation for understanding microbial ecological competition and dynamic balance, but also offers promising avenues for regulating pathogenic bacteria and developing innovative disease control strategies.

Zingiber officinale rhizome is widely cultivated in the central region of Sudan (Gezira) and data on the biological properties of this variety grown in the Sudan climate is scarce. The study aims to comprehensively analyze the antibacterial, antioxidant, phytochemical, and GC-MS properties of Zingiber officinale (Ginger rhizome) to explore its potential applications. The in vitro antibacterial assessment of the aqueous extract of the Sudanese ginger revealed moderate activity against Staphylococcus aureus, Salmonella typhi, Pseudomonas aeruginosa, Escherichia coli , and Klebsiella pneumonia, as resulted from the disc-diffusion which ranged between 12.87±0.11 mm to 14.5±0.12 mm at 30 µg/disc. The minimum inhibition concentration ranged between 6.25 to 25 µg/ml, the MBC were between 25 and 50 µg/ml, and the MBC/MIC exhibited its bactericidal effect against all tested bacteria. The phytochemical screening revealed the presence of various chemical constituents such as saponins, flavonoids, glycosides, alkaloids, steroids, terpenoids, and absence of tannins in the Sudanese ginger rhizome. Furthermore, the GC-MS study of ginger rhizome identified 22 chemical compounds with retention times ranging from 7.564 to 17.023 minutes. The identification of 22 chemical compounds through GC-MS analysis further underscores the prospect of harnessing ginger rhizome for the development of novel medications. The computational analyses showed that ginger compounds bind the 1JIJ and 2QZW with high binding affinities, which reach –9.5 kcal/mol. Ginger compounds also established promising molecular interactions with some key residues, that satisfactorily explain the in vitro results and support the pharmacokinetic and experimental findings. This study lays the groundwork for future research and pharmaceutical exploration aimed at harnessing the beneficial properties of ginger rhizome for medicinal and therapeutic purposes, particularly its antimicrobial potential.

Engineered viral vectors designed to deliver genetic material to specific targets offer significant potential for disease treatment, safer vaccine development, and the creation of novel biochemical research tools. Viral tropism, the specificity of a virus for infecting a particular host, is often modified in recombinant viruses to achieve precise delivery, minimize off-target effects, enhance transduction efficiency, and improve safety. Key factors influencing tropism include surface protein interactions between the virus and host cell, the availability of host cell machinery for viral replication, and the host immune response. This review explores current strategies for modifying the tropism of recombinant viruses by altering their surface proteins. We provide an overview of recent advancements in targeting non-enveloped viruses (Adenovirus and Adeno-associated vi-rus) and enveloped viruses (Retro/Lentivirus, Rabies, Vesicular stomatitis virus, and Herpesvirus) to specific cell types. Additionally, we discuss approaches such as rational design, directed evo-lution, in silico and machine learning-based methods for generating novel AAV variants with desired tropism, and the use of chimeric envelope proteins for pseudotyping enveloped viruses. Finally, we highlight the applications of these advancements, and discuss the challenges and future directions in engineering viral tropism.

Osteoarthritis (OA) is a degenerative joint disease characterized by cartilage degradation and inflammation. This study investigates the therapeutic potential of secretome derived from adipose tissue mesenchymal stem cells (ASCs) in mitigating inflammation and promoting cartilage repair in an in vitro model of OA. Our in vitro model comprised chondrocytes inflamed with TNF. To assess the therapeutic potential of secretome, inflamed chondrocytes were treated with it and concentrations of pro-inflammatory cytokines, metalloproteinases (MMPs) and extracellular matrix markers were measured. In addition, secretome-treated chondrocytes were subject to a microarray analysis to determine which genes were upregulated and which were downregulated. Treating TNF-inflamed chondrocytes with secretome in vitro inhibits the NF-κB pathway, thereby mediating anti-inflammatory and anti-catabolic effects. Additional protective effects of secretome on cartilage are revealed in the inhibition of hypertrophy markers such as RUNX2, COLX and increases in Col2a1 and ACAN production as well as the upregulation of Sox-9. These findings suggest that ASC-derived secretome can effectively reduce inflammation, promote cartilage repair, and maintain chondrocyte phenotype. This study highlights the potential of ASC-derived secretome as a novel, non-cell-based therapeutic approach for OA, offering a promising alternative to current treatments by targeting inflammation and cartilage repair mechanisms.

Several degraded ecosystems worldwide need to attain a stable vegetal cover after mandatory active restoration; thus, plants soil, and soil biodiverse microbes can be taken into consideration for a successful mission. Among environmental and biological variables, interactions with soil and its microorganisms are critical for permanent plant establishment. Thus, advances in the use of microbial management such as inoculation of mycorrhizas and bacteria become a modern aspect of active restoration programs. New strengths have increased worldwide since most plant species, including grasses, associate to mycorrhizae, Thus, a high percent of restored areas, which were degraded due to the increasing mining or conventional agriculture, rest to be monitored. This study shows current reports on active restoration of degraded landscapes as well as the methodology required for a successful ecosystem restoration. Active restoration has been recognized as the best method to restore tropical forests; however, the higher cost of active restoration, which requires planting seedlings, direct seeding, root inoculation of microorganisms or mitigation of disturbances, which aid the recovery process despite monitoring. On the other hand, the spontaneous recovery of trees that colonize and establish in abandoned fields or natural disturbances, named natural regeneration, is less targeted and can require more time to achieve results. We conducted a web of science search of recent published articles on restoration and analyzed them. We confirmed the important role of AMF on root colonization and soil AMF spores as drivers of restoration of main ecosystems according to previous results from restored sites in Brazil. In the restored sites AMF (more than 8 species) was generally found, while in preserved sites dominated Gigaspora, Scutellospora and Acaulospora, restored sites presented relevant AMF richness as well as the preserved sites. Root colonization by AMF in experimental mined sites was high. Vesicles were frequent in restored sites.; however, colonization intensity was generally the same despite treatments. In general, in the control treatments, values of root colonization remain high.

Phosphatidylserine (PS) has significant applications in various sectors, such as the medical and food industries. However, its production relies heavily on phospholipase D (PLD), a crucial tool that is hindered by issues like poor stability and irrecoverability. Immobilization presents itself as an effective solution to overcome these limitations. In this study, magnetic graphene oxide (MGO) modified with amino (-NH₂) were synthesized and utilized for PLD immobilization. The activity of the immobilized PLD (MGO-PLD) reached 3062 U/g_MGO, with a specific activity of 33.9 U/mg_PLD, virtually identical to that of the free PLD. MGO-PLD was utilized to synthesize PS efficiently in a biphasic system. Under optimal conditions, PS yield reached 18.66 g/L, with a conversion rate of 92.8% and a productivity of 3.11 g/L/h. Notably, MGO-PLD retained an impressive PS conversion rate of 77.4% even after seven repetitive usages. Moreover, MGO-PLD displayed enhanced thermal and pH resistance properties compared to free PLD, alongside an augmented storage stability. Post a 8-week preservation at 4°C, its residual activity was able to maintain at 76.3%. This study provides a sustainable and highly efficient pathway for biocatalytic synthesis of PS.

Melanoma, a deadly form of skin cancer, has seen improved survival rates due to advances in diagnosis and treatment, yet the need for further improvement remains critical. Tumor-associated antigens, such as PRAME (Preferentially Expressed Antigen in Melanoma), offer promising avenues for enhanced diagnostic precision, prognostic assessment, and targeted immunotherapy. PRAME, a cancer-testis antigen, is selectively expressed in various cancers, including melanoma, and plays a key role in promoting tumorigenesis through inhibition of retinoic acid signaling, epithelial-to-mesenchymal transition, and immune evasion. This review explores the diagnostic utility of PRAME in distinguishing melanoma from benign nevi, its prognostic value in aggressive melanoma subtypes, and its potential as a therapeutic target in cancer vaccines and adoptive T-cell therapies. While PRAME-targeted therapies face challenges such as tumor heterogeneity and immune suppression, ongoing research aims to overcome these barriers, offering hope for more effective melanoma treatments.

Microalgae and their bioproducts hold significant potential across various applications, for instance, in wastewater remediation, CO2 fixation, and the synthesis of nutraceuticals, pharmaceuticals, and biofuels. The production of these organisms heavily relies upon environmental conditions, including factors like temperature, light, and nutrient supply, which can significantly impact growth. Furthermore, the limitations in nutrients or certain extreme environments can result in productivity increases. As such, microalgae growth systems have become a critical consideration for both research and industry. Both open and closed systems, such as raceway ponds and photobioreactors, respectively, have their own advantages and drawbacks in microalgae cultivation. However, for microalgae growth, photobioreactors may address most concerns, as the system has a lower risk of contamination and can deliver specific growth factors in a controlled manner. This review will summarize the growth and incubation factors of microalgae and the uses of bioreactors in both research and industrial capacities.

Superficial acral fibromyxoma (SAF) is a benign fibromyxoid tumor that presents as a slow-growing nodule on the subungual and periungual areas of the hands and feet. Superficial acral fibromyxoma needs to be better documented in the literature, and thus, increased research and education are required to aid clinicians in recognizing its appearance. To review the current literature on superficial acral fibromyxoma and present its epidemiology, pathogenesis, clinical presentation, diagnosis, treatment, and common differential diagnosis. A literature review was performed on Pubmed and Scopus for case reports, case series, clinical studies, and reviews that described epidemiology, clinical presentation, pathogenesis, histological examination, and treatment of superficial acral fibromyxoma. The gathered literature was analyzed to extract concise information on superficial acral fibromyxoma. Results: Superficial acral fibromyxoma is associated with Rb1 gene deletion and grows as a 0.5 to 5.0-centimeter gelatinous-to-firm nodular mass on periungual or subungual regions of middle-aged males. It is also noted to disrupt nail architecture and cause lytic or erosive lesions of bone. Histopathologically, superficial acral fibromyxoma comprises stellate and spindle cells surrounded by a myxo-collagenous matrix and is strongly positive for CD34, CD99, nestin, and vimentin. Complete surgical excision is the first-line treatment for superficial acral fibromyxoma; however, the tumor has a post-operative recurrence rate of 20-25%. Mohs surgery has been attempted to decrease the rate of recurrence. After excluding other cutaneous neoplasms, superficial acral fibromyxoma should be considered in patients with a periungual or subungual slow-growing mass and other soft tissue. Complete surgical excision can lead to recurrence; therefore, patient follow-up is necessary.

Background: Regular moderate-vigorous intensity exercise is recommended for adults as it can improve longevity and reduce health risks associated with a sedentary lifestyle. However, there are barriers to achieving intense exercise that may be addressed using virtual reality (VR) as a tool to promote exercise intensity and adherence, particularly through visual feedback and competition. The purpose of this work is to compare visual feedback and competition within fully-immersive VR to enhance exercise intensity and user experience of exercise for young adults; and to describe and compare visual attention during each of the conditions. Methods: Young adults (21- 34 years old) bicycled in three 5-minute VR conditions (visual feedback, self-competition and competition against others). Exercise intensity (cycling cadence and % of maximum heart rate) and visual attention (derived from a wearable eye tracking sensor) were measured continuously. User experience was measured by an intrinsic motivation questionnaire, perceived effort and participant preference. A repeated measures ANOVA with paired t-test post-hoc tests was conducted to detect differences between conditions. Results: Participants exercised at a higher intensity and had higher intrinsic motivation in the two competitive conditions compared to visual feedback. Further, participants preferred the competitive conditions and only reached a vigorous exercise intensity during self-competition. Visual exploration was higher in visual feedback compared to self-competition. Conclusion: For young adults bicycling in VR, competition promoted higher exercise intensity and motivation compared to visual feedback.

Plastic pollution is a pressing environmental challenge, necessitating the development of biodegradable alternatives like polyhydroxybutyrate (PHB). This study focuses on optimizing PHB production by Neobacillus niacinii GS1, a bacterium isolated from a municipal dumping site. By utilizing agricultural residues such as corn flour, wheat bran, and peptone as substrates, we aimed to establish an eco-friendly method for biopolymer production, contributing to sustainable waste management and bioplastic innovation. The bacterium was identified using morphological, biochemical, and molecular techniques. The optimization process involved adjusting variables such as inoculum age, inoculum size, incubation time, agitation rate, incubation temperature, pH of the medium, carbon sources and nitrogen sources. Response surface methodology (RSM) was employed to identify optimal conditions, with the highest PHB yield of 61.1% achieved under specific conditions: 37°C, pH 7, and an agitation rate of 150 rpm. These findings underscore the potential of Neobacillus niacinii GS1 in converting agro-industrial residues into valuable biopolymers, promoting sustainable bioplastic production, and advancing waste valorization efforts through the use of eco-friendly materials.

The problem can be stated as over 3 billion choices to improve 14 disease states with 9 optimization goals (some of the optimization goals are diametrically opposed) to improve dyspnea, shortness of breath, fatigability, exercise intolerance, edema, swelling, fluid retention, and arrhythmias. The goal is to increase Left Ventricular Outflow Integral, reduce mitral regurgitation, increase longitudinal conduction velocities and restore synchrony of the septum to the ventricle that needs it the most. A method to accomplish this task for the spectrum of cardiac disease is presented.

Mesenchymal stem cells (MSCs) hold immense therapeutic potential due to their regenerative and immunomodulatory properties. However, to utilize this potential, it is crucial to optimize their in vitro cultivation conditions. Three-dimensional (3D) culture methods using cell-laden hydrogels aim to mimic the physiological microenvironment in vitro, thus preserving MSC biological functionalities. Cellulosic hydrogels are particularly promising due to their biocompatibility, sustainability, and tunability in terms of chemical, morphological, and mechanical properties. This study investigated the impact of (1) two physical crosslinking scenarios for hydrogels derived from anionic cellulose nanofibers (CNF) used to encapsulate adipose-derived MSCs (adMSCs) and (2) physiological culture conditions on the in vitro proliferation, differentiation, and extracellular vesicle (EV) production of these adMSCs. Our results revealed that additional Ca2+-mediated crosslinking, intended to complement self-assembly and gelation of aqueous to-CNF in the adMSC cultivation medium, adversely affected both the mechanical properties of the hydrogel spheres and the growth of the encapsulated cells. However, cultivation under dynamic and hypoxic conditions significantly improved the proliferation and differentiation of the encapsulated adMSCs. Furthermore, we demonstrated that adMSCs in CNF hydrogel spheres exhibited potential for scalable EV production with potent immunosuppressive capacities in a bioreactor system. These findings underscore the importance of physiological culture conditions and the suitability of cellulosic materials for enhancing the therapeutic potential of MSCs. Overall, our study provides valuable insights for optimizing the in vitro cultivation of MSCs for various applications, including tissue engineering, drug testing, and EV-based therapies.

Inherited retinal diseases (IRDs) are clinically complex and genetically heterogeneous visual impairment disorders with varying penetrance and severity. Disease-causing variants in at least 289 nuclear and mitochondrial genes have been implicated in their pathogenesis. In the current study, we performed exome sequencing on a 51 years-old Ashkenazi Jewish patient with non-syndromic retinitis pigmentosa (RP) and identified compound heterozygous variants in the CLRN1 gene: a known pathogenic missense [p.(N48K)] and a novel deep intronic variant c.254-643G&gt;T. A minigene splicing assay that was performed aiming to study the effect of the c.254-643G&gt;T variant on CLRN1 pre mRNA splicing revealed the inclusion of a pseudo-exon that was also reported to be included in the transcript due to an adjacent variant, c.254-649T&gt;G. However, unlike the reported c.254-649T&gt;G variant, c.254-643G&gt;T showed aberrant splicing in a leaky manner, implying that the identified variant is not totally penetrant. The non-syndromic phenotype observed in this index case may be attributed to the leaky nature of this variant, which is causing some normal transcripts to be produced. To conclude, we report on a novel deep intronic variant in CLRN1 causing non-syndromic RP.

The integration of genetic technologies into neurological diagnostics has significantly enhanced our ability to understand and manage neurological disorders. This article discusses the functional analysis of key genetic tests, including microarray analysis, variant sequencing, exome sequencing, RNA sequencing, and whole genome sequencing. Each of these technologies offers unique advantages and challenges, providing a comprehensive toolkit for diagnosing and understanding the genetic basis of neurological conditions. The functional insights gained from these tests not only detect genetic abnormalities but also elucidate their functional consequences, aiding in the diagnosis, prognosis, and management of neurological disorders. As these technologies continue to evolve, they are expected to play an increasingly important role in the field of neurological diagnostics, paving the way for personalized medicine and improved patient outcomes.

Candida albicans, a prominent constituent of the human microbiota, poses a significant health threat due to its opportunistic pathogenicity and adept biofilm-forming capabilities. This research delves into the antifungal attributes and biofilm inhibition potential of Tea Tree Oil (Melaleuca alternifolia) cultivated in Vietnam. Renowned for its bioactive compounds, including terpinen-4-ol, 1,8-cineole, and terpinolene, the essential oil of Melaleuca alternifolia, commonly referred to as tea tree oil (TTO), is scrutinized for its impact on Candida albicans. The study meticulously determines the Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC) of TTO against Candida albicans in both planktonic and biofilm states. Additionally, it assesses the biofilm-forming capacity of Candida 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 Candida 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 results underscore the promising potential of TTO as a potent antifungal agent and biofilm inhibitor against Candida albicans, offering valuable insights for the innovation of novel therapeutic strategies in the realm of fungal infections.

This article delves into the historical development and ramifications of divergent theories that have emerged since the publication of Darwin's seminal work, "On the Origin of Species" in 1856. It traces the evolution of thought from Darwin's revolutionary ideas through the controversial era of eugenics, the groundbreaking discovery of DNA, the elucidation of Mendelian genetics, and the subsequent development of modern genetic theories. The article also explores the critical role of mathematical models, statistics, and data science in advancing genomics research over the last decade. This comprehensive exploration highlights how these theories and methods have fundamentally shaped our understanding of biology, evolution, human genetics, and complex biological systems.

Background: Having a laboratory renal profile for medical imaging examinations requiring contrast media (CM) administration is strongly advised. Creatinine helps identify patients at risk of contrast-induced nephropathy (CIN). GEM® Premier™ ChemSTAT (Werfen) is a point-of-care (POC) analyzer with 12 emergency parameters including creatinine assay. The aim of this study consists to compare ChemSTAT with the central analytical solution of the University Hospital of Clermont-Ferrand and to evaluate the interest of the POC creatinine in the emergency department (ED) in optimizing the flow of patients, especially if CM used. Methods: More than 200 whole blood (WB) samples from the ED were evaluated on the ChemSTAT analyzer. As comparative methods, the plasma aliquots from the same samples were assayed on an Atellica® CH (Siemens Healthineers). The clinical concordance was assessed ac-cording the decision cut-offs of the French Society of Radiology for the risk of CIN. The availability times of biological results between ChemSTAT and the central laboratory were studied. Results: WB results from ChemSTAT correlated well with those from Atellica® CH, except for tCO2 (known bias between Siemens and Cobas Roche methods predicting ChemSTAT values). The results of creatinine assay allow to have identical medical decisions compared to the renal risk cut-offs. The availability of biological results is reduced by 50 min on average with ChemSTAT vs the central laboratory. Computed tomography was performed for 44.7% of patients including the injection of the CM in 68% of cases. For these patients, the availability of creatinine results relative to imaging time is faster with the ChemSTAT on average of 45.2 min. Conclusions: Great analytical and clinical correlations for creatinine assay allow safe iden-tification of patients at risk of CIN and improve patient flow in ED especially for those requiring computed tomography with use of CM.

Biological pre-treatment holds much promise as an innovative and sustainable strategy to improve biogas production from crop residues. In the present study, a novel hot spring cellulolytic microbial consortium (HSCMC) was used to pre-treat crop residues, such as maize stover, wheat straw and soybean straw to improve biogas production by anaerobic digestion (AD). Results indicated that the ash and volatile solids in maize stover were significantly reduced by maximum values of 21.3 % and 69.2 % respectively, after 7 days of pre-treatment over non-treated residues. The maximum decline of 83.9 % in total solid content was reported in wheat straw. The total reducing sugar content was increased by maximum yields of 60.9 %, 96.3 % and 84.7 % in maize stover, wheat straw and soybean straw, respectively. Furthermore, the HSCMC consortium enhanced the cumulative methane yield of wheat straw, maize stover and soybean straw by 50.6 %, 50.2 % and 56.6 %, respectively. Increased methane yield from pre-treated crop residues implies that this method could be an economically viable option for large-scale valorization of the AD process.

The use of C-type natriuretic peptide (CNP) in the interaction with the oocyte and in the temporary postponement of spontaneous meiosis resumption has already been well described. However, its action in pre-implantation developmental stage embryos remains to be understood. Thus, our study aimed to detect the presence of the canonical CNP receptor (natriuretic peptide receptor, NPR2) in germinal vesicle (GV), metaphase II (MII), presumptive zygotes (PZ), morula (MO), and blastocyst (BL) stage embryos and, later, to observe possible modulations on the embryos when co-cultured with CNP. In the Experiment I, we detected and quantified the NPR2 on the above-mentioned embryo stages. Following, Experiment II, intended to test different concentrations (100, 200, or 400 nM of CNP) at different times of inclusion in the in vitro culture (IVC; inclusion from the beginning, i.e. day 1, or from day 5). In Experiment III, it was used 400 nM of CNP on day 1 (D1) in the IVC, which did no demonstrate to be embryotoxic, and it showed potentially promising results in blastocyst production rate, when compared to the control. Thus, we analyzed the embryonic development rates of bovine embryos (D7) and hatching kinetics (D7, D8, and D9). Subsequently, morula and blastocyst were collected and evaluated for transcript abundance to competence and quality (apoptosis, oxidative stress, proliferation and differentiation) and lipid metabolism. Differences with probabilities less than P < 0.05, and/or Foldchange (FC) ˃1.5 were considered significant. We demonstrate the presence of NPR2 until the blastocyst development stage when there was a significant decrease in membrane receptors. There was no statistical difference in production rate after co-culture with 400 nM CNP. However, when we evaluated the abundance of morula transcripts, there was an upregulated transcription in ADCY6 (P=0.057), and downregulated transcripts, in BMP15 (P= 0.013), ACAT1 (P= 0.040), CASP3 (P=0.082). In addition, a total of 12 transcriptions in morula presented variation FC ˃1.5. In blastocysts, the treatment with CNP induced upregulation in BID, CASP3, SOX2, HSPA5 transcripts, and downregulation in BDNF, NLRP5, ELOVL1, ELOVL4, IGFBP4, and FDX1 transcripts (FC >1.5). Thus, our study identified and quantified the presence of NPR2 in bovine pre-implantation embryos. Furthermore, 400 nM of CNP in IVC, a concentration not previously described in the literature, modulated some transcripts related to embryonic metabolism and it was not embryotoxic morphologically.

Malta produces approximately 15,000 tons of bluefin tuna every year, by-products are estimated to count for 55% of the harvest. Maltese tuna farmers established a company that processes these by-products into fish meal and oil that are sold to the pet food industry. In this study the logistic of tuna farming and management in Malta was analysed and the environmental impact of the activity was estimated. The biomass quality and the preservation state of tuna head and liver was assessed. The head had the highest ash content (1.68±0.02%) while the liver had the highest protein content (16.18±0.02%) and the highest crude fat, (34.87±0.76%). The highest concentration of oil was found in the head’s skin (50.07±0.84%). The predominant saturated fatty acids (SFAs) were c14:0, c16:0 and c18:0. Monounsaturated FAs (MUFAs) were the major fatty acids present in all samples, with the highest values in the tuna head. The oxidation levels found in the by-products upon arrival at the processing plant doesn’t compromise their quality. No histamine formation was predicted. Some oxidation was observed in the oil after extraction. The Life Cycle Assessment estimated that the environmental score of the activity is 105pts per ton of fishmeal and 25kg of oil produced (2.82 tCO2 eq). It is concluded that the processing of tuna by-products in Malta is a sustainable alternative to traditional disposal methods and their quality is high enough to be used as a source of proteins and lipids in farmed-animals feed.

This comprehensive overview of the historical milestones in cell culture underscores key breakthroughs that have shaped the field over time. It begins with Wilhelm Roux's seminal experiments in the 1880s, followed by the pioneering efforts of Ross Granville Harrison, who initiated groundbreaking experiments that fundamentally shaped the landscape of cell culture in the early 20th century. Carrel's influential contributions, notably the immortalization of chicken heart cells, mark a significant advancement in cell culture techniques. Holtfreter, Moscona, and Leighton introduced innovations such as 3D cell culture, laying the groundwork for further exploration. The advent of induced pluripotent stem cells by Takahashi and Yamanaka in 2006 was revolutionary, enabling the reprogramming of differentiated cells into a pluripotent state. Since then, recent innovations include spheroids, organoids, and organ-on-a-chip technologies, aiming to mimic the structure and function of tissues and organs in vitro, pushing the boundaries of biological modeling and disease understanding. In this review, we overview the history of cell culture shedding light on the main discoveries, pitfalls and hurdles that were overcome during the transition from 2D to 3D cell culture techniques. Finally, we discussed the future directions for cell culture research that may accelerate the development of more effective and personalized treatments.

Myeloperoxidase (MPO) is an enzyme that contains a heme group, found mostly in neutrophils and in small amounts in monocytes and plays a major role in their anti-microbial activity. However, excessive levels of MPO have been linked to various disorders and identified as a major cause of tissue destruction. Inhibiting its activity can reduce the severity and extent of tissue damage. Over activity of MPO during chronic inflammation has been shown to be involved in tumorigenesis by inducing a hyper-mutagenic environment through oxidant interaction with DNA, causing DNA modification. Vascular endothelium is one of the most important targets of MPO and high levels have been associated with increased rates of cardiomyopathy, ischemic stroke, heart failure, myocardial infarction, and atrial fibrillation. Therefore, it may be considered a therapeutic target in the treatment of cardiovascular disorders. MPO also participates in the pathogenesis of neurodegenerative diseases. For example, an increase in MPO levels has been observed in the brain tissue of patients with Alzheimer's, Multiple sclerosis (MS), and Parkinson's diseases. In Alzheimer's disease, active MPO is mostly found in the location of beta amyloids and microglia. Therefore, targeting MPO may be a potential treatment and prevention strategy for neurological disorders. This review will discuss MPO's physiological and pathological role in cancer, cardiovascular, and neurological disorders.

The landscape of clinical management for metastatic melanoma (MM) and other solid tumors has been modernized by the advent of immune checkpoint inhibitors (ICI), including programmed cell death-1 (PD-1), programmed cell death-ligand 1 (PD-L1), and cytotoxic T lymphocyte antigen 4 (CTLA-4) inhibitors. While these agents demonstrate efficacy in suppressing tumor growth, they also lead to immune-related adverse events (irAEs), resulting in exacerbation of autoimmune diseases such as Rheumatoid arthritis (RA), Ulcerative colitis (UC), and Crohn’s disease (CD). The immune checkpoint inhibitors offer promising advancements in the treatment of melanoma and other cancers, but they also present significant challenges related to irAEs and autoimmune diseases. Ongoing research is crucial to better understand these challenges and develop strategies for mitigating adverse effects while maximizing therapeutic benefits. In this manuscript, we addressed this challenge using network-based approaches by constructing and analyzing molecular and signaling networks associated with tumor-immune cross-talk.. Our analysis revealed that IL6 is the key regulator responsible for irAEs during ICI therapies. Furthermore, we conducted an integrative network and molecular-level analysis, including virtual screening, of drug libraries, such as the Collection of Open Natural Products (COCONUT) and the Zinc15 FDA-approved library, to identify potential IL-6 inhibitors.. Subsequently, the compound amprenavir was identified as the best molecule that may disrupt essential interactions between IL6 and IL6R responsible for initiating signaling cascades underlying irAEs in ICI therapies.

This study investigates the potential inhibitory role of Amentoflavone against the Ebola virus, with a focus on the viral protein VP24 structure. Molecular docking simulations were conducted to assess the binding affinity and interaction between Amentoflavone and the VP24 protein. The results revealed promising docking scores, suggesting favorable binding of Amentoflavone to the VP24 structure. These findings provide valuable insights into the potential therapeutic efficacy of Amentoflavone against Ebola virus infection by targeting specific viral proteins. Further experimental validation is warranted to confirm these docking results and elucidate the mechanism of action underlying Amentoflavone's antiviral activity. This study contributes to the ongoing efforts to develop novel therapeutic strategies for combating Ebola virus disease.

We have previously demonstrated that receptor-interacting serine threonine kinase 1 (RIPK1) is expressed in hair follicle and regulates hair cycle. RIPK1 inhibitors also accelerated the telogen-to-anagen transition and elongated the anagen period in the mouse model. Here, we first investigated the involvement of RIPK1 in alopecia areata (AA). mRNA and protein expression of RIPK1 was increased in the skin of AA mouse model. Single cell RNA sequencing and immunohistochemistry showed that RIPK1 is highly increased in dendritic cells (DCs) and CD8+ T cells. RIPK1 inhibitors (i.e., Necrostatin-1s and GSK2982772) delayed the onset of AA in mouse model, and reduced the DCs and CD8+ T cells in AA skin. RIPK1 inhibitors also increased the hair length in the mouse hair organ culture mimicking AA. Collectively, these results suggest that RIPK1 is involved in AA onset via modulating immune cells, and RIPK1 inhibitors could prevent AA onset.

Several studies have shown promising treatments for type 1 diabetes mellitus (T1DM) using developing gene therapy methods. This review article focuses on summarizing possible new treatment options and outlining their pros and cons, since there is no cure for T1DM as of yet. New treatments involve islet and stem cell transplantation, genetic vaccination, and immunological precursor cell-mediated therapy. Since most of these methods are still in their early stages, efficacy in human trials of this type of treatment is still largely unknown, however, researchers are hopeful after numerous successes in trials with non-obese diabetic (NOD) mice.

Antibiotic resistance of cyanobacteria has been a global threat to public health. The widespread presence of cyanobacteria in aquatic environments exposes them to antibiotic contamination. The cyanobacteria are also in direct contact with pathogenic bacteria containing Antibiotic-Resistance Genes (ARGs) that gives them these characteristics. The study aims to the presence of some ARGs in locally isolated cyanobacteria species, Spirulina laxa, Chroococcus minutes, Oscillatoria princeps, Oscillatoria proteus, Oscillatoria terebriformis, Lyngbya epiphytica, and to compare the presence of these genes in two pathogenic bacteria, Escherichia coli and Klebsiella pneumoniae. The results revealed the presence of The ampicillin (Ap) and erythromycin (Em) resistance genes were detected in five algal samples. Meanwhile, the chloramphenicol (Cm) and gentamicin (Gm) resistance genes were apparent in only two species. Genes encoding resistance towards kanamycin (Km) and spectinomycin (Sp) were recorded in three specimens. The results also documented that E. coli possessed the resistance genes for four antibiotics: Ampicillin (Ap), Erythromycin (Em), Gentamicin (Gm), and Kanamycin (Km), whereas K. pneumoniae was resistant towards three anti-biotics: Ampicillin (Ap), Gentamicin (Gm), and Kanamycin (Km). The results show that there are a match in antibiotic resistance genes in both cyanobacteria and pathogenic bacteria. suggesting the possibility that cyanobacteria could acquire ARGs from the environment through horizontal gene transfer. Thus, freshwater cyanobacteria may play an important role in the prevalence of ARGs in their environment.

The field of neurology has undergone a profound transformation over the past two decades, shifting from traditional clinical and pathological approaches to a more genetically informed perspective. This essay explores the drastic turn in neurology from classical disciplines to neurogenetics, examining the factors that have contributed to this shift and the implications for clinical practice and research. The traditional approach to neurology, grounded in clinical examination, neuroanatomy, and neuropathology, has been effective in diagnosing and treating many conditions but has had limitations in understanding the etiology of complex and rare neurological disorders. The emergence of neurogenetics has been driven by advancements in genetic technologies, such as next-generation sequencing (NGS) and genome-wide association studies (GWAS), as well as the recognition of the genetic basis of many neurological disorders. This shift has led to more accurate diagnoses, targeted treatments, and a deeper understanding of the molecular mechanisms underlying neurological disorders. However, challenges remain, including the complexity of genetic architecture and the ethical implications of genetic testing. The future of neurogenetics holds great promise for improving the diagnosis, treatment, and prevention of neurological conditions, with new technologies and interdisciplinary approaches poised to further advance the field.

Mosquitoes are responsible for the transmission of numerous pathogens, including Plasmodium parasites, arboviruses and filarial worms. They pose a significant risk to public health with over 200 million cases of malaria per annum and approximately four billion people at risk of arthropod-borne viruses (arboviruses). Mosquito populations are geographically expanding into temperate regions and their distribution is predicted to continue increasing. Mosquito symbionts, including fungi, bacteria and viruses, have desirable traits for mosquito disease control including spreading horizontally and vertically through mosquito populations and potentially colonising multiple important vector species. Paratransgenesis, genetic modification of mosquito symbionts with effectors to target the pathogen rather than the vector, is a promising strategy to prevent the spread of mosquito-borne diseases. A variety of effectors can be expressed but venom toxins are excellent effector candidates because they are target specific, potent, and, stable. However, the only toxins to be explored in mosquito paratransgenesis to date are Scorpine and mutated phospholipase A2. To enhance the scope, effectiveness, and durability of paratransgenesis, an expanded arsenal of effectors is required. This review discusses other potential toxins effectors for future paratransgenesis studies based on prior in vitro and in vivo antiparasitic and antiviral studies and highlights the need for further research and investment in this area. In terms of mosquito-borne diseases, paratransgenesis strategies have been developed to target Plasmodium. We postulate the potential to apply this principle to target arboviruses using antiviral toxin effectors.

Mitochondria, crucial intracellular organelles, are central to energy metabolism, signal transduc-tion, apoptosis, calcium homeostasis, and a myriad of other biological processes, making them a focal point in diverse research fields. The capacity to fluorescently labeling and visually track mitochondria is crucial for understanding their biological roles. We present Mulberrin-Cy3, a novel small molecule fluorescent probe that selectively labels mitochondria in animal cells, in-cluding cancer cells, with relative ease. This protocol details the synthesis of Mulberrin-Cy3 and its use for visualizing mitochondria in living cells. The synthesis is straightforward and time-efficient, and the labeling method is more accessible than traditional approaches, providing a cost-effective option for mitochondrial visualization at room temperature. The labeling is rapid, with effective labeling achieved within 5 minutes of incubation. The fluorescent signal is stable and brighter, offering a significant advantage over existing methods. Mulberrin-Cy3 represents a promising mitochondrial labeling compound, providing researchers with a novel experimental tool to explore the complex biological functions of mitochondria. This innovation has the potential to significantly advance our comprehension of mitochondrial dynamics and their role in cellular health and disease.

Background: Existing human cell immortalization methods made the cells obtain oncogenesis phenotype and/or caused the cells gain and/or lose chromosomes. Immortalized normal human T cells lines provide critical in vitro models for basic research and therapeutic products development.Methods: We utilized a CRISPR/Cas9 system to replace a single copy of the exon 2 of the cell cycle inhibitor gene CDKN2A (encoding p16 and p14 proteins) with a single copy of human telomerase reverse transcriptase (hTERT) to immortalize human primary CD8⁺ T cells (hCD8⁺T-TERT). By using Cas9 protein and low donor DNA copies/cell, we successfully immortalized hCD8⁺T cells with a single copy of hTERT transgene, which also avoided uncontrolled insertion of Cas9 gene and guide RNA vector.Results: Human primary CD8+ cells were immortalized and expanded more than 2.6 × 10⁷ times. Characterization of the cells revealed that the immortalized CD8⁺ T-TERT cells retained most of the cell surface markers and normal karyotype. The CD8⁺ T-TERT cells also retained the dependence of IL-2 and CD3/CD28 activator for survival and expansion.Conclusion: we established a stable immortalized CD8⁺ T cell line that had a phenotype consistent with T cells and will be a useful research model.

Somatic embryogenesis (SE) provides alternative methodologies for the propagation of grapevine (Vitis spp.) cultivars, conservation of its germplasm resources and for crop improvement. In this review, the current state of knowledge regarding grapevine SE as applied to these technologies is presented, with a focus on the benefits, challenges, and limitations of this method. The paper provides a comprehensive overview of the different steps involved in the grapevine SE process, including callus induction, maintenance of embryogenic cultures, and the production of plantlets. Additionally, the review explores development of high-health plant material through SE, the molecular and biochemical mechanisms underlying SE, including the regulation of gene expression, hormone signalling pathways, and metabolic pathways as well as its use in crop improvement programmes. The review concludes by highlighting the future directions for grapevine SE research, including the development of new and improved protocols, the integration of SE with other plant tissue culture techniques, and the application of SE for the production of elite grapevine cultivars, for the conservation of endangered grapevine species as well as for cultivars with unique traits that are valuable for breeding programs.

Ethiopia is a country that is in the process of development. Unfortunately, the management and utilization of watershed resources in this region are inadequate. This has resulted in a decrease in the quality of essential resources like soil, water, plants, and animals. Our research delved into the intricate biophysical conditions, conservation practices, and key issues surrounding watersheds along with their root causes and current strategies. The study was particularly focused on understanding the reality and different approaches employed in watershed management practices at the Gullele Botanical Garden located in Addis Ababa. We gathered information through interviews with experts and technicians, direct on-site observations, and official records from the garden's administration office. The gathered data was meticulously analyzed both qualitatively and quantitatively using Arc Map 10.3 to gain a comprehensive understanding of the area. Our findings revealed that the main objective of watershed management in areas surrounding restricted forest sites, Dam A, and Dam B was centered on conserving soil and water resources. The study emphasized the importance of selecting appropriate technologies for effective water and soil conservation practices. Data collection was done using purposive and random sampling methods, drawing information from a variety of sources such as the Gullele Botanical Garden office, literature, research reports, student theses, and both published and unpublished organizational documents. Despite these efforts, our study highlighted inefficiencies in the institutions responsible for watershed management. In particular, the maintenance of stone bunds, gullies, and gabions was not conducted adequately, leading to project deficits. Moving forward, we recommend raising awareness, enhancing training programs, and strengthening law enforcement and watershed policies to transform the knowledge, skills, and attitudes of technicians and staff involved in watershed management techniques for the better.

AI integration is becoming more important in today’s world and could potentially be very useful in the life sciences and biotechnology. This study evaluates the effectiveness of AI systems in teaching molecular cloning techniques to newcomers in molecular biology, focusing on a comparison between Gemini Advanced (powered by Gemini Ultra 1.0) and ChatGPT (GPT-4o). Molecular or DNA cloning is a lab technique for creating multiple copies of a specific DNA segment. It involves isolating the DNA fragment, inserting it into a vector, and introducing it into a host cell, typically bacteria, which then generates numerous copies of the DNA. This essential technique underpins genetic engineering, allowing for gene study, protein expression, and the creation of genetically modified organisms. The primary objective of this study was to assess whether individuals with limited laboratory experience could effectively acquire a specific molecular biology skill set solely through an AI system, rather than relying on the more traditional ‘learning by shadowing’ approach. Participants with limited lab experience completed these tasks using both Gemini and GPT-4o, and evaluated on quality of instructions, conciseness, scientific accuracy, and user satisfaction. Results indicated that both AI systems demonstrated strengths in delivering clear and comprehensive instructions, but in different ways. Gemini, although more interactive with images and diagrams lacked directness and had a harder time keeping past steps in its “memory”. On the other hand, GPT-4o was much more direct and concise in the steps listed and had more accurate results akin to what is performed in the lab. This study underscores the potential of AI-driven tools to simplify complex biotechnological procedures, making them accessible for learners at all levels.

Conventional biochemical methods for studying cellular signaling cascades have relied on destructive cell disruption. In contrast, live cell imaging of fluorescent-tagged transfected proteins offers a non-invasive approach to understanding signal transduction events. One strategy involves monitoring the phosphorylation-dependent shuttling of a fluorescent-labeled kinase between the nucleus and cytoplasm using nuclear localization, export signals, or both. This paper introduces a simple method to visualize intracellular signal transduction in live cells by exploring the translocation properties of PKC from the cytoplasm to the membrane. We fused bait protein to PKC, allowing the bait (RFP-labeled) and target (GFP-labeled) proteins to co-translocate from the cytoplasm to the membrane. However, in non-interacting protein pairs, only the bait protein was translocated to the plasma membrane. To verify our approach, we examined the Raf–MEK–ERK signaling cascade (ERK pathway) and directly observed the Raf1/MEK2 interaction. We also identified ternary complexes containing the KSR1 scaffold protein. Additionally, the MEK/ERK interaction was dependent on the exogenous expression of KSR1.

Phasins are proteins, which are located on the surface of Polyhydroxybutyrate (P3HB) granules and that carry out activities that affect the polymer metabolism. This study aimed to evaluate the effect of inactivating phasins PhbP2 or PhbP3 on the growth rate, production and molecular mass of P3HB in cultures under low or high oxygen transfer rates (OTR). The results reveal that under high OTRₘₐₓ conditions (between 8.1 and 8.9 mmol L⁻¹ h⁻¹ depending on the strain evaluated), the absence of phasins PhbP2 and PhbP3 resulted in a strong negative effect on the growth rate. In contrast, the rates of specific oxygen consumption increased in the case of cultures with strains where phasins were absent. This behavior was not observed under conditions at a low oxygen transfer rate (3.9 ± 0.71 mol L⁻¹ h⁻¹); where both growth and oxygen consumption were the same for the different strains evaluated. It was observed, at high OTRₘₐₓ that the absence of phasin PhbP3 harms the P3HB synthesis, finding a decrease of 30 % in the polymer production at the end of cultivation; however, the molecular weight of the polymer remained constant during the course of cultivation. In summary, it can be concluded that both growth and polymer synthesis were affected when phasin PhbP3 was absent from the P3HB granule, especially at high OTRₘₐₓ.

Inflammation and immune cell dysfunction are critical facilitators of endometriosis pathophysiology. The diverse cellular fractions in the endometriotic microenvironment are represented mainly by glands and stroma, which are infiltrated by immune cells, including macrophages. Macrophages are renowned for stimulating lesion growth, vascularization, and innervation and contributing to pain in the disorder. By combining macrophages and endometriotic cells, we determined if resveratrol and its natural analogs can target the immune dysregulation and oxidative imbalance in endometriosis. We studied the effect of analyzed compounds on the expression of key inflammatory and oxidative stress markers, cytokines release, and ROS production by applying q-PCR, ELISA, Cytometric Beads Array, and multiplexed fluorogenic staining and flow cytometry analysis with bioimaging. The results showed that endometriosis-related macrophages treated with resveratrol or its derivatives have impaired expression of pro-inflammatory markers such as IL6, IL8, IL1B, TNF, CCL2, CXCL10, and PTGS2. This study also reported a pronounced antioxidant effect of the compounds by reducing the accumulation of free radicals in macrophages, which correlated with the transcript activation of detoxification enzymes (SOD, GPx). Considering the harmful macrophage action in endometriosis, we described the prospects for potential therapy based on regulating the immune microenvironment and reducing the accumulation of free radicals.

Managing plant diseases caused by phytopathogenic fungi, like anthracnose caused by Colletotrichum species, is a challenge. Different methods are being sought to identify compounds with antibiotic properties. The Trichoderma strains represent a source of novel molecules with antifungal properties, including Volatile Organic Compounds (VOCs), the production of which is influenced by the medium’s nutrient content. In this study, we assessed the VOCs produced in dual confrontation systems performed in two culture media by Trichoderma strains (T. atroviride IMI206040, T. asperellum T1 and T3, and Trichoderma sp. T2) against Colletotrichum acutatum. We analyze the VOCs profiles using gas chromatography coupled with mass spectrometry. The Luria Bertani medium (LB) stimulated VOCs production with antifungal properties in most systems. We determined the 2-pentyl furan, dimethyl disulfide, and α-phellandrene antifungal activity in vitro. The equimolar mixture of those VOCs (250 µM) generated 14% C. acutatum diametral growth inhibition. The infective ability and the disease severity caused by the mycelium exposed to the VOCs mixture significantly diminished on strawberry leaves. The application of those VOCs as biofumigants could contribute to managing anthracnose. The use of LB represents a feasible strategy to identify VOCs with antifungal properties even between Trichoderma strains belonging to the same species.

Medicago sativa L. (Alfalfa) is a globally recognized forage legume that has recently gained attention for its high protein content, making it suitable for both human and animal consumption. However, due to its perennial nature and autotetraploid genetics, conventional plant breeding requires a longer timeframe compared to other crops. Therefore, genetic engineering offers a faster route for trait modification and improvement. Here, we describe a protocol for achieving efficient transient gene expression in alfalfa through genetic transformation with the Agrobacterium tumefaciens pCAMBIA1304 vector. This vector contains the reporter genes β-glucuronidase (GUS) and green fluorescent protein (GFP), along with a selectable hygromycin B phosphotransferase gene, all driven by the CaMV 35s promoter. Various transformation parameters—such as different explant types, leaf ages, leaf sizes, wounding types, bacterial concentrations (OD600nm), tissue preculture periods, incubation periods, co-cultivation periods, and different concentrations of acetosyringone, silver nitrate, and calcium chloride—were optimized using 3-week-old plantlets raised in vitro. Results were attained from data based on the semi-quantitative observation of the percentage and number of GUS spots on different days of agro-infection in alfalfa explants. The highest percentage (76.2%) of GUS-positive alfalfa leaf explants was observed after 3 days of agroinfection at a bacterial concentration of 0.6, 2 days of preculture, 30 min of infection time, 150 µM acetosyringone, 4 mM calcium chloride, and 75 µM silver nitrate. The transient expression of genes of interest was confirmed via histochemical GUS and GFP assays. The results based on transient reporter gene expression suggest that various factors influence T-DNA delivery in the Agrobacterium-mediated transformation of alfalfa.

A genome represents the complete collection of genetic material in an organism or cell. This genetic information is encoded in nucleic acids, which can be either single or double-stranded and arranged in linear or circular forms. DNA, the most common type of genetic material, consists of just four nucleotides: adenine, guanine, thymine, and cytosine. These nucleotides are organized in a double helix structure, a discovery made by Watson and Crick. The sequence of these nucleotides forms the basic blueprint of a gene or genome. The process of determining this sequence is known as genome sequencing. The field of genome sequencing has evolved dramatically over time, advancing from sequencing short DNA fragments to analyzing millions of base pairs. Initially, efforts were focused on determining the sequence of individual genes, but now whole genome sequencing is both rapid and widely accessible. Recent improvements in sequencing technology emphasize faster, more accurate results, lower costs, and better data analysis. Sanger sequencing, which became the gold standard for DNA sequencing for about thirty years, was instrumental in completing various genome sequences, including the human genome. It laid the groundwork for newer sequencing methods. Today, these newer methods, known collectively as “Next-Generation Sequencing” (NGS), include several generations of technology. Second-generation methods, such as Illumina, Pyrosequencing, ABI/SOLiD, and Ion Torrent Sequencing, and third-generation methods like PacBio and Helicos Sequencing, have advanced the field further. Fourth-generation technology, such as Nanopore sequencing, is also emerging. Most modern research now relies on NGS for analyzing biological sequences, and understanding these technologies offers insight into current methods and future developments.

Advancements in biological and medical science are intricately linked to the biological central dogma. In recent years, gene editing techniques, especially CRISPR/Cas systems, have emerged as powerful tools for modifying genetic information, supplementing the central dogma and holding significant promise for disease diagnosis and treatment. Extensive research has been conducted on the continuously evolving CRISPR/Cas systems, particularly in relation to challenging diseases, such as cancer and HIV infection. Consequently, the integration of CRISPR/Cas-based techniques with contemporary medical approaches and therapies is anticipated to greatly enhance healthcare outcomes for human. This review begins with a brief overview of the discovery of the CRISPR/Cas system. Subsequently, using CRISPR/Cas9 as an example, a clear description of the classical molecular mechanism underlying the CRISPR/Cas system was given. Additionally, the development of the CRISPR/Cas system and its applications in gene therapy and high-sensitivity disease diagnosis were discussed. Furthermore, we address the prospects for clinical applications of CRISPR/Cas-based gene therapy, highlighting the ethical consideration associated with altering genetic information. This brief review aims to enhance understanding of the CRISPR/Cas macromolecular system and provide insight into the potential of genetic macromolecular drugs for therapeutic purposes.

Wheat straw (WS) is a promising substrate for biogas production by anaerobic digestion (AD) due to its high carbohydrate content. An estimated 0.603 million t yr-1 of WS are generated from wheat production systems in South Africa. This is equivalent to an energy potential of 11 PJ. Despite, WS is still undervalued as a bioenergy resource in South Africa due to its structural complexity and low nitrogen content. WS disposal methods, such as use in livestock bedding, burning and burying into the soil, inter alia, are not sustainable and may contribute to global warming and climate change. Commercialization of the AD of WS needs to be further developed and promoted. Pre-treatment (i.e. physical, chemical, biological and hybrid methods) and anaerobic co-digestion (AcoD) are novel strategies that can support the conversion of WS into biogas and other value-added products. Current and future research should focus on optimizing pre-treatment and AcoD conditions towards industrialization of WS into valuable products. This paper focuses on the potential use of WS for biogas production by AD in South Africa. The aim is to create information that will promote research and development, and encourage policy makers and stakeholder participation and investment in WS biogas technology.

Six solvent fractions isolated from flower bud and leaf ethanolic extracts of Cleistocalyx operculatus were analyzed for their phytochemical contents including phenolics, flavonoids, saponins, tannins, and alkaloids. Antioxidant activities were measured using the ABTS, DPPH, and FRAP assays. The results showed that flower bud aqueous fraction (BAF) and leaf aqueous fraction (LAF) rich in phenolic content (768.18 and 490.74 mg GAE/g dry extract, respectively) exhibited significantly higher antioxidant activities than other fractions. Flower bud hexane fraction (BHF) had remarkably high flavonoid and saponin contents (134.77 mg QE/g and 153.33 mg OA/g dry extract, respectively), followed by that of leaf hexane fraction (LHF) (76.54 mg QE/g and 88.25 mg OA/g dry extract, respectively). BHF and LHF were found to have extremely high antibacterial activity against two H. pylori strains ATCC 51932 and 43504 (MICs of 125 µg/ml). Interestingly, DMC (2’,4’-Dihydroxy-6’-methoxy-3’,5’-dimethylchalcone) isolated from BHF displayed greater antibacterial activity against the bacterial strains (MICs of 25–50 µg/ml) than those of the fractions. In addition, DMC presented potent inhibitory effects on H. pylori-urease (IC50 of 3.2 µg/ml) and α-amylase (IC50 of 83.80 µg/ml), but no inhibition against α-glucosidase. It was also demonstrated that DMC showed its pronounced inhibitory effects on urease activity and biofilm formation of H. pylori, and could increase the membrane permeability of the bacterial cells. Scanning electron micrographs depict that BHF and DMC had strong effects on the cell shape and significantly induced distortion and damage of the cell membrane. The fractions and DMC showed no significant toxicity to four tested human cell lines. Efforts to reduce antibiotic use indicate the need for further researches of the flower buds and DMC as potential products to prevent or treat gastric H. pylori infections.

Pigments are chemical compounds that impart color to materials via absorption, reflection, and refraction. Traditional sources of natural pigments include plant and insect tissues; however, microorganisms such as bacteria, yeasts, algae, and filamentous fungi also hold potential for pigment production. In this study, we investigated pigment production in 20 isolates from our in-house Actinobacteria strain collection, obtained from the banks of the Guaviare and Arauca Rivers in Colombia. Isolates were identified using 16S rRNA gene sequencing. We evaluated different culture media and determined their various bioactivities, including antioxidant, antibacterial, and cytotoxic properties. Promising pigmented hydroalcoholic extracts exhibiting the above-mentioned properties were identified and their chemical compositions were examined using LC-MS. Our findings highlighted a growth medium-dependent effect, as pigment production, coloration, and bioactivity outcomes were influenced by growth media. These results demonstrate the significant potential of Streptomyces isolates as sources of bioactive pigments for diverse applications.

We developed a protein to rapidly and accurately diagnose Chagas disease, a life-threatening illness identified by the WHO as a critical worldwide public health risk. Complicating the current health situation are serological test limitations that contribute to most infected persons being unaware of their condition and therefore untreated. To improve diagnostic testing, we developed an immunological mimic of the etiological agent, Trypanosoma cruzi, by combining ten pathogen specific epitopes within the beta-barrel protein structure of Thermal Green Protein. The resulting multi-epitope protein, DxCruziV3, displayed high specificity and sensitivity as the antibody capture reagent in an ELISA platform with an analytical sensitivity that exceeds WHO recommendations. Within an immunochromatographic platform, DxCruziV3 showed excellent performance for the point of application diagnosis in a region endemic for multiple diseases, the municipality of Barcelos in the state of Amazonas, Brazil. In total, 167 individuals were rapidly tested using whole blood from a finger stick. As recommended by the Brazilian Ministry of Health, venous blood samples were laboratory tested by conventional assays for comparison. Test results suggest utilizing DxCruziV3 in different assay platforms can confidently diagnosis chronic infections by T. cruzi. Rapid and more accurate results will benefit everyone but will have the most noticeable impact in resource-limited rural areas where the disease is endemic.

The application of three-dimensional (3D) printing/bioprinting technologies and cell therapies has garnered significant attention due to their potential in the field of regenerative medicine. This paper aims to provide a comprehensive overview of 3D printing/bioprinting technology and cell therapies, highlighting their results in diverse medical applications, while also discussing the capabilities and limitations of their combined use. The synergistic combination of 3D printing and cellular therapies has been recognised as a promising and innovative approach, and it is expected that these technologies will progressively assume a crucial role in the treatment of various diseases and conditions in the foreseeable future. This review concludes with a forward-looking perspective on the future impact of these technologies, highlighting their potential to revolutionize regenerative medicine through enhanced tissue repair and organ replacement strategies.

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.

The medicinal herb Centaurium erythraea is well-known for its natural ability to treat digestive issues like dyspepsia and liver insufficiency by promoting the secretions of the liver and esophagus. This study aimed to analyze the effectiveness of Centaurium erythraea extracts in treating diabetes. Decoction and Soxhlet procedures were used to examine the extracts, with water serving as the solvent. The extracts were tested for antioxidant capacity, antimicrobial effects, and inhibitory effects on alpha-amylase activity. In comparison to the aqueous extract, the decocted extract had higher levels of polyphenols, total tannins, and anthocyanins, as well as more effective antioxidant capacities. Due to its reduction of alpha-amylase activity and enteric absorption of glucose, Centaurium erythraea has also shown considerable anti-hyperglycemic action.

The immune system is central in controlling acute hepatitis B infection and in patients resolving chronic hepatitis B (CHB). Given that 221 million (75%) of CHB patients reside in low- and middle-income countries, developing a vaccine with therapeutic properties represents a rational and cost-effective approach more than a romantic endeavor. This review systematically analyses the critical variables related to the safety, efficacy, and effectiveness of CHB treatments. HeberNasvac's experience is revisited to address the challenges and potentialities of therapeutic vaccines, as well as the current roadblocks in research and development, registration, and large-scale implementation.

Continuous efforts are required to find ways to protect crop production against the toxicity of petroleum hydrocarbons, such as diesel, contamination of soils. There is a need for identification of candidate plants that are tolerant to diesel toxicity that might also have the potential for remediation of diesel-contaminated soils. In this study, petunia, a popular ornamental plant and a model experimental plant in research on phytoremediation of environmental pollutants, was used to evaluate a novel method for rapidly assessing diesel toxicity based on the tolerance of shoots generated through in vitro plamt cell culture selection. Petunia shoot lines (L1 to L4) regenerated from diesel-treated callus were compared with those from non-diesel-treated callus (control). Significant morphological differences were observed among the tested lines and control, notably with L1 and L4 showing superior growth. In particular, L4 exhibited remarkable adaptability, with increased root development and microbial counts in a diesel-contaminated potting mix, suggesting that the shoots exhibited enhanced tolerance to diesel exposure. Here, this rapid bioassay has been shown to effectively identify plants with varying levels of tolerance to diesel toxicity and could therefore assist accelerated selection of superior plants for phytoremediation. Further research is needed to understand the genetic and physiological mechanisms underlying tolerance traits, with potential applications beyond petunias to other environmentally significant plants.

Soluble inorganic pyrophosphatase (sPPase), a pyrophosphate hydrolase, has been shown to play an important role in several physiological processes such as plant growth and development, metabolic activities, and response to abiotic stresses, but sPPase has been less studied in woody plants. In order to preliminarily verify whether the soluble inorganic pyrophosphatase in Eucommia ulmoides Oliver (E. ulmoides) has a role in resistance to drought stress, we amplified the EuSIP5 (E. ulmoides soluble inorganic pyrophosphatase 5) cDNA sequence from E. ulmoides by RT-PCR. In order to study the drought resistance of the Eucommia ulmoides soluble inorganic pyrophosphatase 5 (EuSIP5) gene, the cDNA sequence of the EuSIP5 gene was amplified from Eucommia ulmoides Oliver by RT-PCR. Bioinformatic analysis indicated that EuSIP5 might be an unstable amphipathic protein, mainly located in the cytoplasm. E. ulmoides, the highest expression of the EuSIP5 gene was found in male leaves and pericarp from April to October, and in leaves of E. ulmoides in July and September. During drought, the expression of EuSIP5 in seedling E. ulmoides leaves was significantly higher than that of the control. An overexpression vector harboring the EuSIP5 was constructed and introduced into the tobacco (Nicotiana tabacum cv. Xanthi). We found that during drought stress, the appearance of wilting was delayed by 4 days in transgenic tobacco plants (OE) compared with wild type plants (WT), and the expression of drought-related genes DET2, CYP85A1, P5CS, ERF1 and F-box was found to be increased in transgenic tobacco leaves, as well as the enzyme activities of the protective enzymes: peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) were significantly higher than those of the WT, and the malondialdehyde (MDA) content was lower than the WT. These results indicated that the introduction of EuSIP5 gene into tobacco increased the expression of drought-related genes, enhanced the anti-membrane oxidation ability of tobacco, and reduced the oxidative stress damage, thus improving the drought resistance of tobacco.

Comparative analysis of mortality rates between public and private healthcare systems remains a critical yet complex area of study. This article synthesizes findings from extensive academic literature and empirical studies to examine the relationship between healthcare system type and patient outcomes across various countries. By focusing on mortality rates as a key indicator, we elucidate the nuanced impacts of public versus private healthcare delivery. Our analysis reveals the multifaceted challenges inherent in both systems and their implications for patient survival. We conclude by identifying crucial gaps in current research and proposing policy recommendations to optimize healthcare outcomes in diverse global contexts.This revised abstract:Starts with a strong opening statement that immediately captures the topic's importance. Clarifies the article's methodology (synthesizing existing literature and studies).Specifies the focus on mortality rates as the primary outcome measure.Hints at the complexity of the findings, avoiding oversimplification.Ends with a clear statement about the article's contributions to both research and policy.

The ovary is the major determinant of female reproductive health. Ovarian functions are mainly related to the primordial follicle pool, which is gradually loss with aging. Ovarian aging and reproductive dysfunctions share oxidative stress as common underlying mechanism. ROS signaling is essential for normal ovarian processes yet can contribute to various ovarian disorders when disrupted. Therefore, balance in the redox system is crucial for proper ovarian functions. In the present study, by focusing on mRNAs and ncRNAs described in the ovary and taking into account only validated ncRNA interactions, we built up an ovarian antioxidant ceRNA network, namely OvAnOx ceRNA, composed of 5 mRNAs (SOD1, SOD2, CAT, PRDX3, GR), 10 miRNAs and 5 lncRNAs (XIST, FGD5-AS1, MALAT1, NEAT1, SNHG1). Our bioinformatic analysis indicated that the components of the OvAnOx ceRNA not only contribute to antioxidant defence, but are also involved in other ovarian functions. Indeed, antioxidant enzymes encoded by mRNA of OvAnOx ceRNA operate within a regulatory network that impacts ovarian reserve, follicle dynamics, and oocyte maturation in normal and pathological conditions. The OvAnOx ceRNA represents a promising tool to unravel the complex dialogue between redox potential and ovarian signalling pathways involved in reproductive health, aging and diseases.

Background; Cell-released extracellular vesicles (EVs) acting as 'metabolically and proteolytically active machines,' show potential in metabolomic and proteomic analysis of serum EVs. Despite diverse challenges, post-isolation omics characterization EVs offers crucial insights for effective analysis; (2) Methods: The research, involved children with Congenital Zika Syndrome, utilizing mass spectrometry for proteomics and GC-MS for metabolite identification. Vesicles were isolated using Izon qEV columns, quantified, and characterized by NTA and TEM. Data analysis employed Cytoescape/String and MetaboAnalyst, revealing variations in metabolic and proteomic profiles among groups through PCA and volcano plots. Proteins and Metabolite set enrichment analysis provided biologically meaningful patterns to enriched metabolites; (3) Results: Using molecular exclusion chromatography, the EVs were characterized, revealing size variations. Protein analysis identified 13 significantly altered proteins, including upregulated (e.g., AOM8Q6 - IGLC7) and downregulated (e.g., Q8TD86 - CALML6) ones. Metabolite analysis indicated involvement in the PI3K-AKT-mTOR pathway and suggested a role in Angiotensin inhibition in CZS+. Upstream of mTOR, Akt is the central signaling molecule in the PI3K pathway and plays critical roles in brain development as well as synaptic plasticity important for Zika Virus. The study provides insights into molecular mechanisms associated with CZS; (4) Conclusions: The study pinpointed valuable possible biomarkers, specifically proteins and metabolites, in Zika virus (ZIKV) infection. It stresses the necessity for broader investigations with advanced techniques to uncover molecular targets, potentially advancing pharmacological strategies.

Three-dimensional (3D) models with improved biomimicry are essential to reduce animal experimentation and drive innovation in tissue engineering. In the current study, we investigate the use of alginate-based materials as polymeric inks for 3D bioprinting of osteogenic models using human bone marrow stem/stromal cells (hBMSCs). A composite bioink incorporating alginate, nano-hydroxyapatite (nHA), type I collagen (Col) and hBMSCs was developed and for extrusion-based printing. Rheological tests perfomed on crosslinked hydrogels confirm the formation of solid-like structures consistently indicating a higher storage modulus compared to the loss modulus. The swelling behaviour analysis showed that the addition of Col and nHA into an alginate matrix can enhance the swelling rate of the resulting composite hydrogels, which maximizes cell proliferation within the structure. The LIVE/DEAD assay outcomes demonstrate that the inclusion of nHA and Col did not detrimentally affect the viability of hBMSCs over seven days post-printing. PrestoBlueTM revealed a higher hBMSCs viability in the alginate-nHA-Col hydrogel compared to the remaining groups. Gene expression analysis revealed that alginate-nHA-col bioink favoured a higher expression of osteogenic markers, including secreted phosphoprotein-1 (SPP1) and collagen type 1 alpha 2 chain (COL1A2), in hBMSCs after 14 days, indicating the pro-osteogenic differentiation potential of the hydrogel. This study demonstrates that the incorporation of nHA and Col into alginate enhances osteogenic potential and provides a bioprinted model to systematically study osteogenesis and the early stages of tissue maturation in vitro.

Aflatoxin contamination (AC) increases as the severity of drought stress increase in peanut. Identifying drought-tolerant (DT) genotypes with resistance to Aspergillus. flavus in semi-arid tropics may aid in development of peanuts that minimizing aflatoxin contamination. The goal is to identify the DT genotypes and resistant to aflatoxin contamination. The experiments curried out at ICRISAT Sahelian Center on fifty-five genotypes assessed in adjacent intermittent water-stressed (WS) from 60th day after sowing to maturity and well-watered (WW) conditions in a randomized complete block design. The yields and components, incidence of A. flavus colonisation, aflatoxin contamination and the seed coat total pthe olyphenol (SCTPP) were estimated. Water deficit reduced pod yield, seed yield, and haulm yield up to 19.49%, 27.24% and 22.07% respectively and increased the immature pods plant-1, (IMPN) and aflatoxin contamination up to 67.16% and 54.95%. The genotypes ICG 2106, ICG 311, ICG 4684, ICG 4543, and ICG 1415 maintained high yield under WS, less of IMPN and the lowest AC variation between WW and WS. The drought tolerant genotypes showed the relationship with aflatoxin resistance and SCTPP (r2=0.80; r2=0.82) under WW and WS. These findings can be used to select the genotype that combine drought tolerance and minimizing aflatoxin contamination.

Ewing sarcoma (ES) is an aggressive bone and soft-tissue pediatric cancer. High vitronectin (VN) expression has been associated with poor prognosis in other cancers, and we aimed to determine the utility of this extracellular matrix glycoprotein as a biomarker of aggressiveness in ES. Silk-fibroin plus gelatin-tyramine hydrogels (HGs) were fabricated with and without cross-linked VN and cultivated with A673 and PDX73 ES cell lines for two and three weeks. VN secretion to culture media was assessed using ELISA. Morphometric analysis was applied for phenotypic characterization. VN release to culture media was higher in 3D-models than in monolayer cultures, and intracellular, intercellular and pericluster presence was also observed. A673-HGs showed lower density of clusters but a proportion of larger clusters than PDX73-HGs, which presented low cluster circularity. The cluster density of A673-HGs without added VN was higher than with added VN, and slightly lower in the case of PDX73-HGs. Furthermore, a culture time of three weeks provided no benefits in cluster growth compared to two weeks, especially in A673 -HGs. These advances in 3D modeling and digital quantification pave the way for future studies in ES and other cancers to deepen understanding about intra- and intercellular heterogeneity and anti-adhesion VN therapies.

Myelination, the process of forming a myelin sheath around nerve fibers, is critical not only during early development but also throughout adulthood. This article explores the multifaceted impacts of myelination on the adult brain, highlighting its importance in cognitive functions, neurological health, and overall brain efficiency. In adults, efficient myelination enhances information processing speed, supports memory consolidation, and contributes to cognitive reserve, potentially delaying the onset of neurodegenerative diseases. Myelination ensures synchronized neural activity and energy-efficient brain function, playing a key role in complex cognitive tasks and coordination. Despite a slower rate of plasticity compared to early development, the adult brain retains the ability to adapt and learn, with myelination facilitating these processes. This article also draws parallels between myelination in the brain and learning curves in deep learning Perceptrons, emphasizing how reduced stimulation and experience can significantly slow down cognitive and learning processes. Additionally, the clinical implications of myelination are discussed, including its role in multiple sclerosis and age-related cognitive decline, along with strategies to maintain and enhance myelin health through lifestyle choices and emerging pharmacological interventions. Understanding the ongoing significance of myelination in the adult brain, and its comparison to artificial neural networks, is crucial for developing interventions to support cognitive health and mitigate neurological disorders.

As shown previously, Rhodamine 6G (R6G) and 4-nitro-2,1,3-benzoxadiazole (NBD) linked through a spacer molecule spermidine (spd), R6G-spd-NBD, produce a fluorescent probe with pH-sensitive FRET effect, that can be useful in a variety of diagnostic applications. Specifically, cancer cells can be spotted due to a local decrease in pH (Warburg effect). Here we extend this approach to infectious diseases – namely, leishmaniasis, brucellosis, and tuberculosis, are difficult to treat largely because of their localization inside macrophages. R6G-spd-NBD offers an opportunity to detect such bacteria and potentially deliver therapeutic target to treat them. The micellar form of the R6G-spd-NBD probe (chitosan or heparin grafted with lipoic acid residues, Chit-LA and Hep-LA, respectively) was obtained to improve the pH sensitivity in the desired range (5.5–7.5) and enable the selective targeting of bacterial cells, thus improving CLSM imaging. According to AFM (atomic force microscopy) data, the micelles containing NBD-spd-R6G have a compact and highly spherical shape, with a diameter ranging from 70 to 110 nm. The micellar form of the R6G-spd-NBD further improves absorption and penetration into bacteria, including those located inside macrophages. Due to negative charge of the surface, bacteria absorb positively charged R6G-spd-NBD, and even more so in the micellar form, when chitosan derivatives are used to form the polymeric micelle. Additionally, macrophages’ lysosomes can be easily distinguished due to their acidic pH. Confocal laser scanning microscopy was used to visualize samples of macrophages cells containing absorbed bacteria. The micellar formulation showed a significant selectivity to model E. coli vs Lactobacillus bacterial cells, and R6G-spd-NBD, being a mild bactericide, clears over 50% E.coli in conditions where Lactobacillus remains almost unaffected. Taken together, our data indicates that R6G-spd-NBD, as well as similar compounds, can have value not only for diagnostic, but also for theranostic applications.