Protein Synthesis Research Articles

Construction of Bacillus subtilis chassis strain with enhanced α-amylase expression capability based on CRISPRi screening

Bacillus subtilis has been widely used in the expression of recombinant proteins due to its food safe and powerful secretion characteristic, but the current production level cannot meet the increasing industrial needs. To enhance the production of recombinant protein, we first screened target key genes that are directly or indirectly involved in protein synthesis, using CRISPRi technology targeting the whole genome, with industrial valuable Bacillus stearothermophilus α-amylase as the model protein. Then the screened key genes were combined, yielding a chassis strain that owning enhanced protein expression capability. Following overlaying molecular chaperone GroES/L and peptidoglycan glycosyltransferase PonA, α-amylase activity reached 102,893 U/mL in a 3-L fermenter, the highest level reported till now. Finally, transcriptome analysis showed that the enhanced recombinant expression may be due to more rational allocation of energy and resources. These strategies can be well implicated in engineering other microbial cell factories for higher industrial production.

Characterization of high protein soybean using mass spectrometry based proteomic and metabolomic analyses.

We have created several high protein (HPBL) and low protein breeding lines (LPBL) by crossing between NC-Raleigh and PI407228. The NC-Raleigh is a low protein cultivar widely grown in North Carolina with excellent yield potential and a high protein wild soybean (Glycine soja) with low yield. We analyzed the F6 generation of this cross for protein and oil compositions using near-infrared spectroscopy (NIR). Two soybean lines HPBL and LPBL were selected for biochemical analysis using mass spectrometry-based proteomic and metabolomic techniques. We identified 4,538 proteins and 267 metabolites in these lines. The HPBL showed the enrichment of proteins associated with protein synthesis, processing, and storage in addition to higher content of primary and secondary metabolites. The mapping of metabolites on the global metabolic pathways revealed the increased protein content in the HPBL that may have resulted from increased arginine content routed from citrulline-argininosuccinic acid. This information will be useful for breeders and biotechnologists to produce a value-added soybean trait.

Transcriptome and molecular evidence of HvMORF8 conferring drought-tolerance in barley

Drought is one of the most devastating abiotic stresses worldwide, which severely limits crop yield. Tibetan wild barley is a treasure trove of useful genes for crop improvement including drought tolerance. Here, we detected large-scale changes of gene expression in response to drought stress with a substantial difference among contrasting Tibetan barley genotypes XZ5 (drought-tolerant), XZ54 (drought-sensitive) and cv. Tadmor (drought-tolerant). Drought stress led to upregulations of 142 genes involved in transcription, metabolism, protein synthesis, stress defense, transport and signal transduction in XZ5, but those genes were down-regulated or unchanged in XZ54 and Tadmor. We identified and functionally characterized a novel multiple organellar RNA editing factors 8 (HvMORF8), which was up-regulated by drought stress in XZ5, but unchanged in XZ54 and Tadmor under drought stress. Phylogenetic analysis showed that orthologues of HvMORF8 can be traced back to the closest gymnosperm species such as Cycas micholitzii, implicating a potential evolutionary origin for MORF8 from a common ancestor in early seed plants. Virus-induced HvMORF8 silencing in XZ5 led to hypersensitivity to drought stress, demonstrating it is a positive regulator of drought tolerance in barley. RNA sequencing of BSMV:HvMORF8 and control plants reveals that silencing of HvMORF8 suppresses genes involved in osmolytes transport, cell wall modification and antioxidants, resulting in water metabolism disorder and overaccumulation of reactive oxygen species (ROS) under drought stress. Therefore, we propose HvMORF8-mediated regulatory drought tolerance mechanisms at transcriptomic level in XZ5, providing new insight into the genetic basis of plastid RNA editing function of HvMORF8 for barley drought tolerance.

At what cost? The impactof bacteriophage resistance on the growth kinetics and protein synthesis of Escherichia coli.

Cost of bacteriophage resistance (COR) is important in explaining processes of diversification and coexistence in microbial communities. COR can be expressed in different traits, and the lack of universally applicable methods to measure fitness trade-offs makes COR challenging to study. Due to its fundamental role in growth, we explored protein synthesis as a target for quantifying COR. In this study, the growth kinetics of three genome-sequenced strains of phage-resistant Escherichia coli, along with the phage-susceptible wild-type, were characterized over a range of glucose concentrations. Bioorthogonal non-canonical amino acid tagging (BONCAT) was used to track differences in protein synthetic activity between the wild-type and phage-resistant E. coli. Two of the resistant strains, with different levels of phage susceptibility, showed mucoid phenotypes corresponding with mutations in genes associated with the Rcs phosphorelay. These mucoid isolates, however, had reduced growth rates and potentially lower protein synthetic activity. Another resistant isolate with a different mutational profile maintained the same growth rate as the wild-type and showed increased BONCAT fluorescence, but its yield was lower. Together, these findings present different patterns of trade-offs resulting from the phage-induced mutations and demonstrate the potential applicability of BONCAT as a tool for measuring COR.

New mutation in the β-spectrin gene in hereditary spherocytosis: A case report

In patients with recurrent anemia, jaundice, and splenomegaly, a thorough assessment of family history and peripheral blood smears is crucial for diagnosing hereditary spherocytosis (HS). Furthermore, gene sequencing can enhance diagnostic accuracy and facilitate the investigation of disease mechanisms at the molecular level. In this report, we present a case of HS caused by a heterozygous nonsense mutation in the SPTB gene, along with a family history of this specific mutation. A 35-year-old man was evaluated for jaundice and splenomegaly, which he had experienced since childhood. Blood tests revealed anemia, reticulocytosis, elevated indirect bilirubin levels, and an increased percentage of spherical red blood cells in the peripheral blood. His family history indicated that both his father and daughter exhibited similar clinical manifestations. Subsequently, genetic sequencing confirmed that the patient, along with his father and daughter, shared the heterozygous missense mutation c.155G > A (p.Arg52Gln) in the SPTB gene, which is absent in public population and animal sequence databases. Structural prediction analysis of the protein suggests that this mutation may lead to instability of SPTB mRNA, thereby affecting the synthesis of the spectrin protein and the integrity of the red blood cell membrane skeleton. Further research is needed to clarify the exact relationship between this mutation and the occurrence of HS.

Designing a Secretory form of RTX-A as an Anticancer Toxin: An In Silico Approach.

Cancer is a leading cause of death and a significant public health issue worldwide. Standard treatment methods such as chemotherapy, radiotherapy, and surgery are only sometimes effective. Therefore, new therapeutic approaches are needed for cancer treatment. Sea anemone actinoporins are pore-forming toxins (PFTs) with membranolytic activities. RTX-A is a type of PFT that interacts with membrane phospholipids, resulting in pore formation. The synthesis of recombinant proteins in a secretory form has several advantages, including protein solubility and easy purification. In this study, we aimed to discover suitable signal peptides for producing RTX-A in Bacillus subtilis in a secretory form. Signal peptides were selected from the Signal Peptide Web Server. The probability and secretion pathways of the selected signal peptides were evaluated using the SignalP server. ProtParam and Protein-sol were used to predict the physico-chemical properties and solubility. AlgPred was used to predict the allergenicity of RTX-A linked to suitable signal peptides. Non-allergenic, stable, and soluble signal peptides fused to proteins were chosen, and their secondary and tertiary structures were predicted using GOR IV and I-TASSER, respectively. The PROCHECK server performed the validation of 3D structures. According to bioinformatics analysis, the fusion forms of OSMY_ECOLI and MALE_ECOLI linked to RTX-A were identified as suitable signal peptides. The final proteins with signal peptides were stable, soluble, and non-allergenic for the human body. Moreover, they had appropriate secondary and tertiary structures. The signal above peptides appears ideal for rationalizing secretory and soluble RTX-A. Therefore, the signal peptides found in this study should be further investigated through experimental researches and patents.

Advances in the study of LNPs for mRNA delivery and clinical applications.

Messenger ribonucleic acid (mRNA) was discovered in 1961 as an intermediary for transferring genetic information from DNA to ribosomes for protein synthesis. The COVID-19 pandemic brought worldwide attention to mRNA vaccines. The emergency use authorization of two COVID-19 mRNA vaccines, BNT162b2 and mRNA-1273, were major achievements in the history of vaccine development. Lipid nanoparticles (LNPs), one of the most superior non-viral delivery vectors available, have made many exciting advances in clinical translation as part of the COVID-19 vaccine and therefore has the potential to accelerate the clinical translation of many gene drugs. In addition, due to these small size, biocompatibility and excellent biodegradability, LNPs can efficiently deliver nucleic acids into cells, which is particularly important for current mRNA therapeutic regimens. LNPs are composed cationic or pH-dependent ionizable lipid bilayer, polyethylene glycol (PEG), phospholipids, and cholesterol, represents an advanced system for the delivery of mRNA vaccines. Furthermore, optimization of these four components constituting the LNPs have demonstrated enhanced vaccine efficacy and diminished adverse effects. The incorporation of biodegradable lipids enhance the biocompatibility of LNPs, thereby improving its potential as an efficacious therapeutic approach for a wide range of challenging and intricate diseases, encompassing infectious diseases, liver disorders, cancer, cardiovascular diseases, cerebrovascular conditions, among others. Consequently, this review aims to furnish the scientific community with the most up-to-date information regarding mRNA vaccines and LNP delivery systems.

Delayed application of water and fertilizer increased wheat yield but did not improve quality parameters

Context or problemWheat is an important food crop for mankind, simultaneous improvement of wheat yield and quality is human most important goal, but they often cannot increase at the same time. Objective or research questionThe effects of irrigation mode, nitrogen application rate and topdressing time on yield and quality of two wheat varieties were studied, and the possibility of synergistic improvement of yield and quality was also explored. MethodsTwo irrigation treatments (conventional border irrigation and micro-sprinkling irrigation) were set up, and three nitrogen application rate treatments (120, 210 and 300kgN ha−1) were set up under each irrigation treatment. The yield formation and quality parameters of wheat were investigated. ResultsThe grain yield, key enzyme activities of protein synthesis, protein content and dough processing quality of the two cultivars increased with the increase of nitrogen application rate. However, when the nitrogen level was higher than 210kgN ha−1, the grain yield and processing quality of winter wheat did not increase further. Under the same nitrogen application rate, compared with conventional border irrigation (CBI), micro-sprinkling irrigation (MSI) significantly increased the grain yield and total protein content, but did not improve the ratio of glutenin to gliadin and dough processing quality parameters. ConclusionsUnder the experimental conditions, the synergistic improvement of wheat yield and quality parameters could be achieved by optimizing nitrogen application rate, and the delayed application of water and fertilizer could further increase wheat grain yield, but did not improve grain quality parameters. Implications or significanceThis study provided theoretical guidance for further improving the grain yield and quality of winter wheat in the North China Plain.

Altered purine and pentose phosphate pathway metabolism in uteroplacental insufficiency-induced intrauterine growth restriction offspring rats impairs intestinal function

BackgroundThis study aimed to identify metabolic alterations in the small intestine of newborn rats with intrauterine growth restriction (IUGR), a condition linked to intestinal dysfunction. MethodsPregnant Sprague Dawley rats underwent bilateral uterine artery ligation on gestational day 17 to induce intrauterine growth restriction or sham surgery. Rat pups were delivered spontaneously on gestational day 22. Small intestine tissues were collected on postnatal days 0 and 7 from offspring. Liquid chromatography-mass spectrometry analysis was performed to investigate untargeted metabolomic profiles. Western blot analysis assessed protein expression of key regulators. ResultsNewborn rats with intrauterine growth restriction exhibited distinct small intestine metabolic profiles compared to controls on postnatal day 0. Notably, significant alterations were observed in purine metabolism, the pentose phosphate pathway, and related pathways. Western blot analysis revealed a decrease expression in transketolase, a key enzyme of the pentose phosphate pathway, suggesting impaired activity of the pentose phosphate pathway. Additionally, decreased expression of tight junction proteins ZO-1 and occludin indicated compromised intestinal barrier function in rats with intrauterine growth restriction. Similar metabolic disruptions persisted on postnatal day 7, with further reductions in tricarboxylic acid cycle intermediates and folate biosynthesis precursors. Interestingly, lysyl-glycine, a protein synthesis marker, was elevated in rats with intrauterine growth restriction. ConclusionsOur findings reveal a distinct metabolic signature in the small intestine of neonatal rats with intrauterine growth restriction, characterized by disruptions in the pentose phosphate pathway, purine metabolism, and energy production pathways. These novel insights suggest potential mechanisms underlying IUGR-associated intestinal dysfunction and impaired growth.

Signaling by neutrophil G protein-coupled receptors that regulate the release of superoxide anions.

In human peripheral blood, the neutrophil granulocytes (neutrophils) are the most abundant white blood cells. These professional phagocytes are rapidly recruited from the bloodstream to inflamed tissues by chemotactic factors that signal danger. Neutrophils, which express many receptors that are members of the large family of G protein-coupled receptors (GPCRs), are critical for the elimination of pathogens and inflammatory insults, as well as for the resolution of inflammation leading to tissue repair. Danger signaling molecular patterns such as the N-formylated peptides that are formed during bacterial and mitochondrial protein synthesis and recognized by formyl peptide receptors (FPRs) and free fatty acids recognized by free fatty acid receptors (FFARs) regulate neutrophil functions. Short peptides and short-chain fatty acids activate FPR1 and FFA2R, respectively, while longer peptides and fatty acids activate FPR2 and GPR84, respectively. The activation profiles of these receptors include the release of reactive oxygen species (ROS) generated by the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Activation of the oxidase and the production of ROS are processes that are regulated by proinflammatory mediators, including tumor necrosis factor α and granulocyte/macrophage colony-stimulating factor. The receptors have signaling and functional similarities, although there are also important differences, not only between the two closely related neutrophil FPRs, but also between the FPRs and the FFARs. In neutrophils, these receptors never walk alone, and additional mechanistic insights into the regulation of the GPCRs and the novel regulatory mechanisms underlying the activation of NADPH oxidase advance our understanding of the role of receptor transactivation in the regulation of inflammatory reactions.

Exploring the dynamics of messenger ribonucleoprotein-mediated translation repression.

Translational control is crucial for well-balanced cellular function and viability of organisms. Different mechanisms have evolved to up- and down-regulate protein synthesis, including 3' untranslated region (UTR)-mediated translation repression. RNA binding proteins or microRNAs interact with regulatory sequence elements located in the 3' UTR and interfere most often with the rate-limiting initiation step of translation. Dysregulation of post-transcriptional gene expression leads to various kinds of diseases, emphasizing the significance of understanding the mechanisms of these processes. So far, only limited mechanistic details about kinetics and dynamics of translation regulation are understood. This mini-review focuses on 3' UTR-mediated translational regulation mechanisms and demonstrates the potential of using single-molecule fluorescence-microscopy for kinetic and dynamic studies of translation regulation in vivo and in vitro.

AtLEC2-Mediated Enhancement of Endoplasmic Reticulum-Targeted Foreign Protein Synthesis in Nicotiana benthamiana Leaves: Insights From Transcriptomic Analysis.

Many proteins used in industrial and pharmaceutical applications are typically synthesized within the secretory pathway. While yeast and mammalian cells have been engineered to enhance the production of endomembrane-targeted proteins, similar strategies in plant cells remain underexplored. This study investigates the potential of arabidopsis leafy cotyledon 2 (AtLEC2), a key regulator of seed development, to enhance the production of proteins targeted to the endoplasmic reticulum (ER) in Nicotiana benthamiana leaves. Through transient expression experiments, we demonstrate that AtLEC2 selectively increases the production of ER-targeted GUS without affecting its cytosolic variant. Moreover, leaves agroinfiltrated with AtLEC2 show a significant increase in ER-GFP accumulation compared to controls lacking AtLEC2. Transcriptomic analysis reveals that AtLEC2 promotes ribosome and chloroplast biogenesis, along with the upregulation of genes involved in photosynthesis, translation, and membrane synthesis. Notably, seed-specific poly(A) binding proteins involved in RNA stability and translation initiation, as well as 3-hydroxy-3-methylglutaryl coenzyme A reductase-linked to ER hypertrophy-are highly upregulated. This study establishes a novel connection between AtLEC2 and the enhancement of ER-targeted foreign protein synthesis, paving the way for innovative strategies in plant cellular engineering.

Chemotherapeutic agents and leucine deprivation induce codon-biased aberrant protein production in cancer.

Messenger RNA (mRNA) translation is a tightly controlled process frequently deregulated in cancer. Key to this deregulation are transfer RNAs (tRNAs), whose expression, processing and post-transcriptional modifications are often altered in cancer to support cellular transformation. In conditions of limiting levels of amino acids, this deregulated control of protein synthesis leads to aberrant protein production in the form of ribosomal frameshifting or misincorporation of non-cognate amino acids. Here, we studied leucine, an essential amino acid coded by six different codons. Surprisingly, we found that leucine deprivation leads to ribosomal stalling and aberrant protein production in various cancer cell types, predominantly at one codon, UUA. Similar effects were observed after treatment with chemotherapeutic agents, implying a shared mechanism controlling the downstream effects on mRNA translation. In both conditions, a limitation in the availability of tRNALeu(UAA) for protein production was shown to be the cause for this dominant effect on UUA codons. The induced aberrant proteins can be processed and immune-presented as neoepitopes and can direct T-cell killing. Altogether, we uncovered a novel mode of interplay between DNA damage, regulation of tRNA availability for mRNA translation and aberrant protein production in cancer that could be exploited for anti-cancer therapy.

Physiological and ecological responses of flue-cured tobacco to field chilling stress: insights from metabolomics and proteomics

IntroductionCurrently, research on tobacco's response to chilling stress is mostly limited to laboratory simulations, where temperature is controlled to study physiological and molecular responses. However, laboratory conditions cannot fully replicate the complex environment of field chilling stress, so conducting research under field conditions is crucial for understanding the multi-level adaptive mechanisms of tobacco to chilling stress in natural environments. MethodsThis study aims to use field trials, starting from physiological responses, combined with proteomics and untargeted metabolomics, to systematically reveal the physiological and biochemical characteristics and key molecular mechanisms of tobacco leaves under chilling stress. It provides new insights into tobacco's adaptation strategies under chilling stress. ResultsThe results showed that (1) chilling stress damages the appearance of tobacco leaves, reduces the chlorophyll content, increases H2O2 and malondialdehyde (MDA) levels in cold-injured tobacco leaves, and damages the plasma membrane system. Although catalase (CAT) activity increases to cope with the accumulation of reactive oxygen species (ROS), the activities of key antioxidant enzymes superoxide dismutase (SOD) and peroxidase (POD) significantly decrease, indicating that the antioxidant system of tobacco leaves fails in environments with sudden temperature drops. (2) Proteomics analysis indicated that 410 differentially expressed proteins were identified in cold-stressed tobacco leaves, with 176 upregulated and 234 downregulated. Tobacco leaves under chilling stress attempt to maintain energy supply and physiological stability by enhancing glycolysis, starch, and sucrose metabolism pathways. Concurrently, chilling stress triggers the expression of proteins related to cell wall reinforcement and antioxidant defense. However, due to impaired ribosomal function, protein synthesis is significantly inhibited, which aggravates damage to photosynthesis and cellular functions. (3) Metabolomics analysis revealed that the differential metabolites in cold-stressed tobacco leaves were mainly enriched in tyrosine metabolism, isoquinoline alkaloid biosynthesis, and fatty acid degradation pathways. This indicates that under chilling stress, tobacco leaves enhance adaptability by regulating energy metabolism, increasing antioxidant capacity, and stabilizing cell membrane structure. ConclusionsTherefore, under chilling stress, tobacco leaves exhibit complex physiological adaptability through multiple regulatory mechanisms involving proteins and metabolites. The research results provide important insights into the metabolic regulatory mechanisms of tobacco in response to extreme environments and also enhance the theoretical foundation for addressing low-temperature stress in practical production.

Use of a Novel Feeding System to Assess the Survival of a Very Stable Mammalian Virus, Porcine Parvovirus, Within Black Soldier Fly (Hermetia illucens) Larvae: A Comparison with Mealworm (Tenebrio molitor) Larvae

Insect larvae production offers the potential for large-scale synthesis of high-quality protein that can be used as feed or food. However, currently, there are limitations on the source of substrates for the insect larvae to use. One concern is the potential survival of animal pathogens within insect larvae if their feed is contaminated. In this study, the survival of a very stable virus, porcine parvovirus (PPV), within mealworm (Tenebrio molitor) and black soldier fly (BSF) (Hermetia illucens) larvae has been analyzed after oral ingestion of the virus. PPV genomic DNA could be readily detected by PCR in both species of larvae up until 9 days post ingestion (DPI), the end of the study period. Furthermore, infection of susceptible PK15 cells by PPV from homogenized mealworm larvae could be detected until at least 3 DPI, using an immunoperoxidase staining method and, up until 9 DPI, with a more sensitive real time PCR assay. Thus, PPV can remain infectious within mealworm larvae during their main growth phase through to their harvesting. However, it may be considered that PPV is exceptional in this respect since it displays unusual stability, e.g., to heat.

A mortality timer based on nucleolar size triggers nucleolar integrity loss and catastrophic genomic instability.

Genome instability is a hallmark of aging, with the highly repetitive ribosomal DNA (rDNA) within the nucleolus being particularly prone to genome instability. Nucleolar enlargement accompanies aging in organisms ranging from yeast to mammals, and treatment with many antiaging interventions results in small nucleoli. Here, we report that an engineered system to reduce nucleolar size robustly extends budding yeast replicative lifespan in a manner independent of protein synthesis rate or rDNA silencing. Instead, when nucleoli expand beyond a size threshold, their biophysical properties change, allowing entry of proteins normally excluded from the nucleolus, including the homologous recombinational repair protein Rad52. This triggers rDNA instability due to aberrant recombination, catastrophic genome instability and imminent death. These results establish that nucleolar expansion is sufficient to drive aging. Moreover, nucleolar expansion beyond a specific size threshold is a mortality timer, as the accompanying disruption of the nucleolar condensate boundary results in catastrophic genome instability that ends replicative lifespan.

Soybean Oil and Protein: Biosynthesis, Regulation and Strategies for Genetic Improvement.

Soybean (Glycine max [L.] Merr.) is one of the world's most important sources of oil and vegetable protein. Much of the energy required for germination and early growth of soybean seeds is stored in fatty acids, mainly as triacylglycerols (TAGs), and the main seed storage proteins are β-conglycinin (7S) and glycinin (11S). Recent research advances have deepened our understanding of the biosynthetic pathways and transcriptional regulatory networks that control fatty acid and protein synthesis in organelles such as the plastid, ribosome and endoplasmic reticulum. Here, we review the composition and biosynthetic pathways of soybean oils and proteins, summarizing the key enzymes and transcription factors that have recently been shown to regulate oil and protein synthesis/metabolism. We then discuss the newest genomic strategies for manipulating these genes to increase the food value of soybeans, highlighting important priorities for future research and genetic improvement of this staple crop.

Rapid Development of Modified Vaccinia Virus Ankara (MVA)-Based Vaccine Candidates Against Marburg Virus Suitable for Clinical Use in Humans

Background/Objectives: Marburg virus (MARV) is the etiological agent of Marburg Virus Disease (MVD), a rare but severe hemorrhagic fever disease with high case fatality rates in humans. Smaller outbreaks have frequently been reported in countries in Africa over the last few years, and confirmed human cases outside Africa are, so far, exclusively imported by returning travelers. Over the previous years, MARV has also spread to non-endemic African countries, demonstrating its potential to cause epidemics. Although MARV-specific vaccines are evaluated in preclinical and clinical research, none have been approved for human use. Modified Vaccinia virus Ankara (MVA), a well-established viral vector used to generate vaccines against emerging pathogens, can deliver multiple antigens and has a remarkable clinical safety and immunogenicity record, further supporting its evaluation as a vaccine against MARV. The rapid availability of safe and effective MVA-MARV vaccine candidates would expand the possibilities of multi-factored intervention strategies in endemic countries. Methods: We have used an optimized methodology to rapidly generate and characterize recombinant MVA candidate vaccines that meet the quality requirements to proceed to human clinical trials. As a proof-of-concept for the optimized methodology, we generated two recombinant MVAs that deliver either the MARV glycoprotein (MVA-MARV-GP) or the MARV nucleoprotein (MVA-MARV-NP). Results: Infections of human cell cultures with recombinant MVA-MARV-GP and MVA-MARV-NP confirmed the efficient synthesis of MARV-GP and MARV-NP proteins in mammalian cells, which are non-permissive for MVA replication. Prime-boost immunizations in C57BL/6J mice readily induced circulating serum antibodies binding to recombinant MARV-GP and MARV-NP proteins. Moreover, the MVA-MARV-candidate vaccines elicited MARV-specific T-cell responses in C57BL/6J mice. Conclusions: We confirmed the suitability of our two backbone viruses MVA-mCherry and MVA-GFP in a proof-of-concept study to rapidly generate candidate vaccines against MARV. However, further studies are warranted to characterize the protective efficacy of these recombinant MVA-MARV vaccines in other preclinical models and to evaluate them as vaccine candidates in humans.

Bridging the Gap: Supplements Strategies from Experimental Research to Clinical Applications in Sarcopenic Obesity

Obesity causes fat accumulation, and sarcopenia causes loss of muscle mass and strength; together, they worsen insulin resistance and accelerate muscle decline, creating a harmful cycle. Some supplements, along with physical exercise, could be remedies for sarcopenic obesity (SO). In this review, we aim to draw a comparison between supplements studied in experimental research and those evaluated in clinical studies for SO. In experimental studies, Sea Buckthorn—in forms such as oil, freeze-dried powder or pomace—has been shown to enhance muscle cell growth, improve gut microbiota, provide hypoglycemic benefits and increase muscle mass by promoting protein synthesis. Increased consumption of Omega-3 fatty acids may play a protective role against SO in women. Melatonin may positively impact obesity and SO by reducing oxidative stress. Elevated irisin levels, such as those observed with vitamin D supplementation, could prevent muscle wasting and fat gain in SO by improving insulin sensitivity and reducing inflammation. There have been many studies highlighting the potential of vitamin D in preventing age related sarcopenia; however, the effect of vitamin D supplementation in SO is under-researched and appears less promising. Future clinical trials using natural supplements hold promise, as these provide multiple beneficial components that may work synergistically to treat SO.

Changes in rectal temperature as a means of assessing heat tolerance and sensitivity in chickens

High ambient temperature and relative humidity significantly affect growth and production performance in poultry. Ability of poultry to regulate their core body temperature relative to the ambient temperature depends on the relative nutrient/energy expenditure in maintenance and performance requirements. We hypothesized that changes in rectal temperature corrected for surface area can be used as a measure of heat tolerance/sensitivity. Rectal temperatures of one hundred mixed sex Ross 308 broiler chickens were measured hourly from 6 AM to 6 PM at 24 days of age. The ambient temperature and relative humidity were also measured hourly for the same 12-h period. Body weights were measured at day 24 and 38 days of age. The temperature-humidity index (THI) increased from 77.5 at 6.00 AM and peaked at 83.5 at 3.00 PM. The average rectal temperature increased from 39.900C at 6.00 AM to about 41.300C at 9.00 AM. Thereafter, the average rectal temperature remained constant until 3.00 PM when it began to decline. At 6.00 PM, the rectal temperature had declined to about 40.70 °C. Evaporative heat loss is affected by surface areas and as a result, rectal temperature was corrected for surface area. The change in rectal temperature corrected for surface area was negatively correlated with body weight gain indicating variability in the response of individual chickens exposed to similar THI. This variability was attributed to heat tolerance. It was hypothesized that mismatch between nutrient and energy supply and the partition of nutrient/energy between maintenance of core body temperature and protein synthesis could be reflected on the differences in heat-tolerance and body weight gain in the chicken population. The genetic basis of differences in rectal temperature changes corrected for surface area could be elucidated as a means of developing thermo-tolerant chickens.