Epigenetic reprogramming of H3K4me3 in adipose-derived stem cells by HFS diet consumption leads to a disturbed transcriptomic profile in adipocytes.

Adipose tissue metabolism is actively involved in the regulation of energy balance. Adipose-derived stem cells (ASCs) play a critical role in maintaining adipose tissue function through their differentiation into mature adipocytes. This study aimed to investigate the impact of an obesogenic environment on the epigenetic landscape of ASCs and its impact on adipocyte differentiation and its metabolic consequences. Our results showed that ASCs from rats on a high-fat sucrose (HFS) diet displayed reduced adipogenic capacity, increased fat accumulation, and formed larger adipocytes compared to the control (C) group. Mitochondrial analysis revealed heightened activity in undifferentiated ASC-HFS, but decreased respiratory and glycolytic capacity in mature adipocytes. The HFS diet significantly altered the H3K4me3 profile in ASCs on genes related to adipogenesis, mitochondrial function, inflammation, and immunomodulation. After differentiation, adipocytes retained H3K4me3 alterations, confirming upregulation of genes associated with inflammatory and immunomodulatory pathways. RNA-seq confirmed the upregulation of genes associated with inflammatory and immunomodulatory pathways in adipocytes. Overall, the HFS diet induced significant epigenetic and transcriptomic changes in ASCs, impairing differentiation and causing dysfunctional adipocyte formation.

Coronavirus Disease 2019: Past, Present, and Future.

Severe acute respiratory syndrome coronavirus 2 is one of the most impactful diseases experienced in the past century. While the official national health emergency concluded in May of 2023, coronavirus disease 2019 (COVID-19) continues to mutate. As the summer of 2023, all countries were experiencing a new surge of cases from the EG.5 Omicron variant. Additionally, a new genetically distinct Omicron descendant BA2.86 had been detected in multiple countries including the United States. This article seeks to offer lessons learned from the pandemic, summarize best evidence for current management of patients with COVID-19, and give insights into future directions with this disease.

Evolutionary and Structural Analysis of PP16 in Viridiplantae.

Members of the phloem protein 16 (PP16) gene family are induced by elicitors in rice and the corresponding proteins from cucurbits, which display RNA binding and intercellular transport activities, are accumulated in phloem sap. These proteins facilitate the movement of protein complexes through the phloem translocation flow and may be involved in the response to water deficit, among other functions. However, there is scant information regarding their function in other plants, including the identification of paralog genes in non-vascular plants and chlorophytes. In the present work, an evolutionary and structural analysis of the PP16 family in green plants (Viridiplantae) was carried out. Data mining in different databases indicated that PP16 likely originated from a larger gene present in an ancestral lineage that gave rise to chlorophytes and multicellular plants. This gene encodes a protein related to synaptotagmin, which is involved in vesicular transport in animal systems, although other members of this family play a role in lipid turnover in endomembranes and organelles. These proteins contain a membrane-binding C2 domain shared with PP16 proteins in vascular plants. In silico analysis of the predicted structure of the PP16 protein family identified several ß-sheets, one α-helix, and intrinsically disordered regions. PP16 may have been originally involved in vesicular trafficking and/or membrane maintenance but specialized in long-distance signaling during the emergence of the plant vascular system.

The Characterization of Melanaphis sacchari Microbiota and Antibiotic Treatment Effect on Insects.

Insects are under constant selective pressure, which has resulted in adaptations to novel niches such as crops. This is the case of the pest Melanaphis sacchari, the sugarcane aphid, native to Africa and currently spreading worldwide. The aphid undergoes successful parthenogenesis, causing important damage to a variety of crops and leading to important economic losses for farmers. A natural M. sacchari population grown in sorghum was studied to identify its microbiome through the sequencing of its 16S rDNA metagenome. A high proportion of Proteobacteria, followed by Firmicutes, Bacteroidetes, and Actinobacteria, was observed. We also detected Wolbachia, which correlates with the asexual reproduction of its host. M. sacchari was challenged in a bioassay with the antibiotics oxytetracycline and streptomycin, resulting in a dose-dependent decay of its survival rate. The possibility of controlling this pest by altering its microbiota is proposed.

Long-Distance Movement of Solanum tuberosum Translationally Controlled Tumor Protein (StTCTP) mRNA.

Long-distance signaling molecules in plants, including different RNA species, play a crucial role in the development and environmental responses. Among these mobile signals, the Translationally Controlled Tumor Protein (TCTP) mRNA is one of the most abundant. TCTP regulates cell-cycle progression and programmed cell death and is involved in responses to abiotic and biotic stress as well as plant regeneration, among other functions. Considering that the ability to induce plant regeneration is linked to a possible role of TCTP in vegetative propagation and asexual reproduction, we analyzed TCTP overexpression in a solanaceous plant model that can reproduce asexually by regeneration from stolons and tubers. Therefore, in this study, the effect of transient expression of Solanum tuberosum TCTP (StTCTP) on tuber development and vegetative propagation was described. StTCTP mRNA was shown to be transported long-distance. Additionally, transient overexpression of StTCTP resulted in sprouts with a greater diameter compared to control plants. Furthermore, the early stages of tuberization were induced compared to control plants, in which only mature tubers were observed. These results suggest a role of TCTP in vegetative propagation and asexual reproduction.

Starting from scratch: Step-by-step development of diagnostic tests for SARS-CoV-2 detection by RT-LAMP.

The pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has affected millions of people worldwide. Public health strategies to reduce viral transmission are based on widespread diagnostic testing to detect and isolate contagious patients. Several reverse transcription (RT)-PCR tests, along with other SARS-CoV-2 diagnostic assays, are available to attempt to cover the global demand. Loop-mediated isothermal amplification (LAMP) based methods have been established as rapid, accurate, point of care diagnostic tests for viral infections; hence, they represent an excellent alternative for SARS-CoV-2 detection. The aim of this study was to develop and describe molecular detection systems for SARS-CoV-2 based on RT-LAMP. Recombinant DNA polymerase from Bacillus stearothermophilus and thermostable engineered reverse transcriptase from Moloney Murine Leukemia Virus were expressed using a prokaryotic system and purified by fast protein liquid chromatography. These enzymes were used to set up fluorometric real time and colorimetric end-point RT-LAMP assays. Several reaction conditions were optimized such as reaction temperature, Tris-HCl concentration, and pH of the diagnostic tests. The key enzymes for RT-LAMP were purified and their enzymatic activity was determined. Standardized reaction conditions for both RT-LAMP assays were 65°C and a Tris-HCl-free buffer at pH 8.8. Colorimetric end-point RT-LAMP assay was successfully used for viral detection from clinical saliva samples with 100% sensitivity and 100% specificity compared to the results obtained by RT-qPCR based diagnostic protocols with Ct values until 30. The developed RT-LAMP diagnostic tests based on purified recombinant enzymes allowed a sensitive and specific detection of the nucleocapsid gene of SARS-CoV-2.

Bioinformatic-based approach for mutagenesis of plant immune Tm-22 receptor to confer resistance against tomato brown rugose fruit virus (ToBRFV).

Nucleotide-binding leucine-rich repeat (NLR) plant immune receptors mediate the recognition and activation of defense signaling pathways in response to intra- and extracellular pathogens. Several NLR such as Tm-2 and Tm-22 have been introgressed into commercial solanaceous varieties to confer protection against different tobamoviruses. Particularly, Tm-22 was used during recent decades to confer resistance against tobacco mosaic virus, tomato mottle mosaic virus and tomato mosaic virus, which recognizes the viral movement protein (MP). However, tomato brown rugose fruit virus(ToBRFV), a novel tobamovirus, can avoid the protection conferred by Tm-22 due to the presence of key substitutions in the MP. The aim of this work was to identify the key amino acid residues involved in the interaction between Tm-22 and ToBRFV MP through bioinformatic analyses, and to identify potential Tm-22 mutations that could generate greater binding affinity. In silico 3D structure prediction, molecular docking, and computational affinity methods were performed. We predicted that R350, H384 and K385 Tm-22 residues are relevant for the interaction with MP, and two mutations (H384W and K385L) were identified as putative sites to increase the affinity of Tm-22 to the MP with the potential elicitation of resistance against ToBRFV.

Reaction Kinetics in the Vermicomposting Process of Peach Waste.

Peach is a fruit cultivated in temperate regions and its use generates waste composed of seeds and skin. Inadequate disposal of this waste generates an environmental impact; therefore, an alternative is to apply a vermicomposting degradation process. In this research, these four laboratory-scale reactors were used: RC (no earthworms), R1, R2, and R3 (50 earthworms each) to get mixtures in the following proportions of peach waste and load material (vegetable waste and eggshell): RC (50%-50%), R1 (50%-50%), R2 (60%-40%), and R3 (40%-60%). In addition, during this process, physicochemical parameters were analyzed (temperature, pH, humidity, total organic carbon (TOC), total nitrogen (TN), and carbon/nitrogen ratio (C/N)). For each mixture, the reaction order and rate constants were determined using mathematical models. After analysis of the reaction kinetics, the results showed that zero- and first-order reactions were best suited for the degradation of this waste in the vermicomposting process. The highest rates of degradation in the mixtures were for RC and R1, which means faster completion of the process, and consequently, smaller dimensions of the facilities necessary for vermicomposting. Thus, this research provides important information for the design of reactors that use similar substrates.

Observing pedestrian-vehicle traffic conflicts in school zones to evaluate the effectiveness of road safety interventions and reduce injuries in Ghana, Vietnam, and Mexico, 2019-2021.

BACKGROUND: Daily more than 3,000 children are injured or killed on the road, often along the school route. Road traffic crashes and resulting injuries are preventable. More can be done to reduce injuries and save lives. Traffic Conflict Techniques (TCTs) are simple methods of collecting observational data to evaluate the effectiveness of road safety interventions through counting and analyzing traffic conflicts. A TCT Toolkit was developed and piloted to analyze pedestrian-vehicle traffic conflicts in school zones in low- and middle-income countries. METHODS: Three non-governmental organizations in Ghana, Vietnam, and Mexico applied three TCTs from the TCT Toolkit to collect traffic conflict data before (pre-intervention) and after (post-intervention) road safety intervention implementation. As the number of traffic conflicts was often less than 100, confidence intervals (CIs) based on gamma distributions were calculated for the traffic conflict rate. Using the calculated traffic conflict rate, the difference between pre- and post-intervention rates was assessed by determining overlap of the CIs. When CIs did not overlap, the difference was said to be statistically significant at the 0.05 level. RESULTS: For each method, results indicated a decrease in traffic conflicts between pre- and post-intervention data collection periods. Pre- and post-intervention traffic conflict rates with non-overlapping CIs demonstrated the results were statistically significant, providing evidence that the road safety interventions were effective. CONCLUSIONS: TCTs are relatively low-cost and simple techniques that provide an opportunity to base road safety improvement decisions on real-world data. TCTs are effective in objectively evaluating road safety interventions and can help decision-makers evaluate strategies for improving road safety, preventing injuries and saving lives.

The NAC Transcription Factor ANAC087 Induces Aerial Rosette Development and Leaf Senescence in Arabidopsis.

CmNACP1 mRNA has been shown to move long distance through the phloem in Cucurbita maxima (pumpkin) and through a graft junction. Whereas the phloem transport of several different mRNAs has been documented in other systems as well, its function remains, for most of these RNAs, largely unknown. To gain insight into the possible role of these RNAs, we searched for the closest homologs of CmNACP1 in Arabidopsis, a model plant much more amenable for analysis. A phylogenetic approach using the predicted NAC domain indicated that ANAC059, ANAC092, ANAC079, ANAC100, ANAC046, and ANAC087 form a single clade with CmNACP1. In the present work, we analyzed the possible function of the ANAC087 gene in more detail. The promoter region of this gene directed expression in the vasculature, and also in trichomes, stem, apexes, and developing flowers which supports the notion that ANAC087 and CmNACP1 are orthologs. Overexpression of the ANAC087 gene induced increased branching in inflorescence stem, and also development of ectopic or aerial rosettes in T1 and T2 plants. Furthermore, overexpression of ANAC087 leads to accelerated leaf senescence in 44 days post-germination (dpg). Interestingly, a similar phenotype was observed in plants expressing the ANAC087 gene upstream region, also showing an increase in ANAC087 transcript levels. Finally, the results shown in this work indicate a role for ANAC087 in leaf senescence and also in rosette development.

Transcriptomic Analysis of the Host Response to Mild and Severe CTV Strains in Naturally Infected Citrus sinensis Orchards.

Citrus tristeza virus (CTV) is an important threat to the global citrus industry, causing severe economic losses worldwide. The disease management strategies are focused on vector control, tree culling, and the use of resistant varieties and rootstocks. Sweet orange (Citrus sinensis) trees showing either severe or mild CTV symptoms have been observed in orchards in Veracruz, Mexico, and were probably caused by different virus strains. To understand these symptomatic differences, transcriptomic analyses were conducted using asymptomatic trees. CTV was confirmed to be associated with infected plants, and mild and severe strains were successfully identified by a polymorphism in the coat protein (CP) encoding gene. RNA-Seq analysis revealed more than 900 significantly differentially expressed genes in response to mild and severe strains, with some overlapping genes. Importantly, multiple sequence reads corresponding to Citrus exocortis viroid and Hop stunt viroid were found in severe symptomatic and asymptomatic trees, but not in plants with mild symptoms. The differential gene expression profiling obtained in this work provides an overview of molecular behavior in naturally CTV-infected trees. This work may contribute to our understanding of citrus-virus interaction in more natural settings, which can help develop strategies for integrated crop management.

Genistein Stimulation of White Adipose Tissue Thermogenesis Is Partially Dependent on GPR30 in Mice.

SCOPE: Genistein increases whole body energy expenditure by stimulating white adipose tissue (WAT) browning and thermogenesis. G-Coupled receptor GPR30 can mediate some actions of genistein, however, it is not known whether it is involved in the activation of WAT-thermogenesis. Thus, the aim of the study is to determine whether genistein activates thermogenesis coupled to an increase in WAT browning and mitochondrial activity, in GPR30+/+ and GPR30-/- mice. METHODS AND RESULTS: GPR30+/+ and GPR30-/- mice are fed control or high fat sucrose diets containing or not genistein for 8 weeks. Body weight and composition, energy expenditure, glucose tolerance, and browning markers in WAT, and oxygen consumption rate, 3', 5'-cyclic adenosine monophosphate (cAMP) concentration and browning markers in adipocytes are evaluated. Genistein consumption reduces body weight and fat mass gain in a different extent in both genotypes, however, energy expenditure is lower in GPR30-/- compared to GPR30+/+ mice, accompanied by a reduction in browning markers, maximal mitochondrial respiration, cAMP concentration, and browning markers in cultured adipocytes from GPR30-/- mice. Genistein improves glucose tolerance in GPR30+/+ , but this is partially observed in GPR30-/- mice. CONCLUSION: The results show that GPR30 partially mediates genistein stimulation of WAT thermogenesis and the improvement of glucose tolerance.

Engineering Macromolecular Trafficking Into the Citrus Vasculature.

The plant vasculature is a central organ for long-distance transport of nutrients and signaling molecules that coordinate vegetative and reproductive processes, and adaptation response mechanisms to biotic and abiotic stress. In angiosperms, the sieve elements are devoid of nuclei, thus depending on the companion cells for the synthesis of RNA and proteins, which constitute some of the systemic signals that coordinate these processes. Massive analysis approaches have identified proteins and RNAs that could function as long-range signals in the phloem translocation stream. The selective translocation of such molecules could occur as ribonucleoprotein complexes. A key molecule facilitating this movement in Cucurbitaceae is the phloem protein CmPP16, which can facilitate the movement of RNA and other proteins into the sieve tube. The CmPP16 ortholog in Citrus CsPP16 was characterized in silico to determine its potential capacity to associate with other mobile proteins and its enrichment in the vascular tissue. The systemic nature of CsPP16 was approached by evaluating its capacity to provide phloem-mobile properties to antimicrobial peptides (AMPs), important in the innate immune defense. The engineering of macromolecular trafficking in the vasculature demonstrated the capacity to mobilize translationally fused peptides into the phloem stream for long-distance transport. The translocation into the phloem of AMPs could mitigate the growth of Candidatus Liberibacter asiaticus, with important implications for crop defense; this system also opens the possibility of translocating other molecules to modulate traits, such as plant growth, defense, and plant productivity.

Development of a droplet digital polymerase chain reaction (ddPCR) assay for the detection of Tomato brown rugose fruit virus (ToBRFV) in tomato and pepper seeds.

In the present study, a droplet digital PCR assay was developed for detection of Tomato brown rugose fruit virus, a new Tobamovirus of tomato and other solanaceous plants, which expands the diagnostic strategies for this pathogen. Candidate reference DNA material was also obtained to be employed as positive control in tomato and pepper samples. Recombinant plasmids encode for ToBRFV coat protein (CP-ToBRFV) gene and Solanum lycopersicum GAPDH fragments, and CP-ToBRFV and Capsicum annuum GAPDH. To our knowledge, this is the first report of ToBRFV detection in tomato and pepper seeds using ddPCR.

Recombinant Antigens Based on Non-Glycosylated Regions from RBD SARS-CoV-2 as Potential Vaccine Candidates against COVID-19.

The Receptor-Binding Domain (RBD) of the Spike (S) protein from Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has glycosylation sites which can limit the production of reliable antigens expressed in prokaryotic platforms, due to glycan-mediated evasion of the host immune response. However, protein regions without glycosylated residues capable of inducing neutralizing antibodies could be useful for antigen production in systems that do not carry the glycosylation machinery. To test this hypothesis, the potential antigens NG06 and NG19, located within the non-glycosylated S-RBD region, were selected and expressed in Escherichia coli, purified by FPLC and employed to determine their immunogenic potential through detection of antibodies in serum from immunized rabbits, mice, and COVID-19 patients. IgG antibodies from sera of COVID-19-recovered patients detected the recombinant antigens NG06 and NG19 (A450 nm = 0.80 ± 0.33; 1.13 ± 0.33; and 0.11 ± 0.08 for and negatives controls, respectively). Also, the purified antigens were able to raise polyclonal antibodies in animal models evoking a strong immune response with neutralizing activity in mice model. This research highlights the usefulness of antigens based on the non-N-glycosylated region of RBD from SARS-CoV-2 for candidate vaccine development.

High Rates of COVID-19 Infection Among Indigenous Maya at a US Safety-Net Health System in California.

Coronavirus disease 2019 (COVID-19) has disproportionately and negatively affected communities of color in the United States, especially Black, Latinx, and Indigenous populations. We report a cluster of COVID-19 cases among the Maya in Alameda County, California, most of whom were misclassified in public health data as nonindigenous Spanish-speaking people. We conducted a retrospective cohort study of all COVID-19 tests performed from April 1 through May 31, 2020, at Alameda Health System. A total of 1561 tests from 1533 patients were performed, with an overall test positivity rate of 17.0% (N = 265). We used the language field from the electronic health record to identify 29 patients as speaking an indigenous Mayan language; by medical record review, we identified 52 additional Maya patients. Maya patients had a test positivity rate of 72.8% as compared with 27.1% (P < .001) for nonindigenous Latinx patients and 8.2% (P < .001) for all other patients. In our sample, 39.6% of patients who had a positive test result for COVID-19 were hospitalized, 11.3% required admission to the intensive care unit (ICU), and 4.9% died of COVID-19. Maya patients had lower rates of hospitalization, ICU admission, and 30-day in-hospital mortality than non-Maya patients. We shared our data with the county health department to inform responses for education, testing, and isolation for Maya patients in Alameda County. Ongoing COVID-19 public health efforts should assess the community prevalence of COVID-19 in the Maya community and other indigenous communities and implement interventions that are linguistically and culturally appropriate.

Racial/ethnic disparities in COVID-19 disease burden & mortality among emergency department patients in a safety net health system.

BACKGROUND: We sought to examine racial and ethnic disparities in test positivity rate and mortality among emergency department (ED) patients tested for COVID-19 within an integrated public health system in Northern California. METHODS: In this retrospective study we analyzed data from patients seen at three EDs and tested for COVID-19 between April 6 through May 4, 2020. The primary outcome was the test positivity rate by race and ethnicity, and the secondary outcome was 30 day in-hospital mortality. We used multivariable logistic regression to examine associations with COVID-19 test positivity. RESULTS: There were 526 patients tested for COVID-19, of whom 95 (18.1%) tested positive. The mean age of patients tested was 54.2 years, 54.7% were male, and 76.1% had at least one medical comorbidity. Black patients accounted for 40.7% of those tested but 16.8% of the positive tests, and Latinx patients accounted for 26.4% of those tested but 58.9% of the positive tests. The test positivity rate among Latinx patients was 40.3% (56/139) compared with 10.1% (39/387) among non-Latinx patients (p < 0.001). Latinx ethnicity was associated with COVID-19 test positivity (adjusted odds ratio 9.6, 95% confidence interval: 3.5-26.0). Mortality among Black patients was higher than non-Black patients (18.7% vs 1.3%, p < 0.001). CONCLUSION: We report a significant disparity in COVID-19 adjusted test positivity rate and crude mortality rate among Latinx and Black patients, respectively. Results from ED-based testing can identify racial and ethnic disparities in COVID-19 testing, test positivity rates, and mortality associated with COVID-19 infection and can be used by health departments to inform policy.

Host-specific loss of sequences of an alfalfa mosaic virus isolate during systemic infection.

Infectivity of an alfalfa mosaic virus (AMV) isolate from Leonotis nepetaefolia in different tomato cultivars was analyzed. Symptoms typical of AMV infection were observed in indicator plants, but not in Flora Dade and Rio Grande tomato cultivars; however, mild symptoms were observed in cv. Rutgers. Furthermore, at least 1 kb of the 3´ segment of RNA 2 and the coat protein gene were missing in systemic leaves of inoculated Rio Grande and Flora Dade plants, while in cv. Rutgers infected with this AMV strain all genomic components were detected. Northern blot analysis of plants infected with the aforementioned AMV isolate confirmed the absence of the CP gene, but suggested rearrangements in both RNA 2 and 3. Factors that may affect differential movement or systemic accumulation of genomic components in multipartite viruses in plants are discussed.

The Diversity Snowball Effect: The Quest to Increase Diversity in Emergency Medicine: A Case Study of Highland's Emergency Medicine Residency Program.

Blacks, Hispanics/Latinos, American Indians, Pacific Islanders, Alaska Natives, and Native Hawaiians make up 33% of the US population. These same groups are underrepresented in medicine. In 2013, the physician workforce was 4.1% black, 4.4% Hispanic/Latino, 0.4% American Indian or Alaska Native, 11.7% Asian, and 48.9% white. Only 9.9% of emergency physicians identify as underrepresented minority (4.5% black, 4.8% Hispanic/Latino, and 0.6% American Indian/Alaska Native). Efforts to increase the number of underrepresented minority physicians are important because previous studies show improved outcomes when the patient and physician share the same racial/ethnic background. Starting in 2006, the faculty at the Highland EM Residency Program in Oakland, CA, began a diversification initiative to increase the number of underrepresented minority residents. The goal was to closely mirror the US population and match 30% underrepresented minorities with each incoming class. After the initiative, there was a 2-fold increase in the number of underrepresented minority residents (from 12% to 27%). This article is a review of the strategies used to diversify the Highland EM Residency Program. Most components can be applied across emergency medicine programs to increase the number of underrepresented minority residents and potentially improve health outcomes for diverse populations.