Artisanal mezcal is produced by the natural fermentation of maguey juice, which frequently results in a process that becomes stuck or is sluggish. Using selected indigenous starter inoculums of Saccharomyces and non-Saccharomyces yeasts is considered beneficial in overcoming these problems and thereby preserving the essence of the artisanal process. In this work, three hundred and four yeast isolates were recovered from 17 distilleries and then grouped by the ARDRA analysis, their restriction profiles were clustered in 15 groups. Four of them included 90% of all isolates, and these were identified using the sequence of the D1/D2 domain of the large-subunit rDNA. Pichia kudriavzevii, Pichia manshurica, Saccharomyces cerevisiae, and Kluyveromyces marxianus were detected as predominant species. Both species belonging to the Pichia genus were detected in 88% of the distilleries, followed by S. cerevisiae (70%) and K. marxianus (50%). In order to evaluate the fermentative capacity, one strain of each species was assessed in a pure and mixed culture in two culture media, filtered maguey juice (MJ) and maguey juice including its bagasse (MJB). Findings demonstrated that non-Saccharomyces yeast presented better growth than that of S. cerevisiae. K. marxianus PA16 was more efficient for ethanol production than S. cerevisiae DI14. It produced 32 g/L of ethanol with a yield of 0.47 g/g and efficient of 90%. While, P. kudriavzevii produced more ethyl acetate (280 mg/L) than the others species. All fermentations were characterized by the presence of isobutyl and isoamyl alcohol. The presence of K. marxianus in a mixed culture, improved the ethanol production and volatile compounds increased using co-cultures.

Clostridioides difficile is the causative bacterium in 15-20% of all antibiotic associated diarrheas. The symptoms associated with C. difficile infection (CDI) are primarily induced by the two large exotoxins TcdA and TcdB. Both toxins enter target cells by receptor-mediated endocytosis. Although different toxin receptors have been identified, it is no valid therapeutic option to prevent receptor endocytosis. Therapeutics, such as neutralizing antibodies, directly targeting both toxins are in development. Interestingly, only the anti-TcdB antibody bezlotoxumab but not the anti-TcdA antibody actoxumab prevented recurrence of CDI in clinical trials. In this work, 31 human antibody fragments against TcdB were selected by antibody phage display from the human naive antibody gene libraries HAL9/10. These antibody fragments were further characterized by in vitro neutralization assays. The epitopes of the neutralizing and non-neutralizing antibody fragments were analyzed by domain mapping, TcdB fragment phage display, and peptide arrays, to identify neutralizing and non-neutralizing epitopes. A new neutralizing epitope within the glucosyltransferase domain of TcdB was identified, providing new insights into the relevance of different toxin regions in respect of neutralization and toxicity.

The objective of this study was to investigate the effects of ohmic heating and water bath heating (WB) on the metabolome of Escherichia coli O157:H7 cells at the same inactivation levels. Compared to low voltage long time ohmic heating (5 V/cm, 8.50 min, LVLT) and WB (5.50 min), the high voltage short time ohmic heating (10 V/cm, 1.75 min, HVST) had much shorter heating time. Compared to the samples of control (CT), there were a total of 213 differential metabolites identified, among them, 73, 78, and 62 were presented in HVST, LVLT, and WB samples, revealing a stronger metabolomic response of E. coli cells to HVST and LVLT than WB. KEGG enrichment analysis indicated that the significantly enriched pathways were biosynthesis and metabolism of amino acids (alanine, arginine, aspartate, and glutamate, etc.), followed by aminoacyl-tRNA biosynthesis among the three treatments. This is the first metabolomic study of E. coli cells in response to ohmic heating and presents an important step toward understanding the mechanism of ohmic heating on microbial inactivation, and can serve as a theoretical basis for better application of ohmic heating in food products.

No-tillage agriculture can sustain productivity and protect the environment. A comprehensive understanding of soil arbuscular mycorrhizal (AM) fungal diversity and soil carbon distribution within aggregate fractions is essential to the evaluation of no-tillage agriculture. The long-term field experiment included two tillage treatments (1) no tillage with straw returned to the soil (NTS), and (2) conventional mouldboard-plowing tillage without straw (CT), and was conducted on the Loess Plateau, north-western China, from October 2009. The soil samples were collected from the surface layer (0-20 cm depth) at the maturation stage of the summer maize (Zea mays L.) for analyzing aggregates separated by the dry-sieving method. The organic carbon content in the bulk soil and different particle size aggregates were measured using the dichromate oxidization method. The species compositions of soil AM fungi were compared by applying high-throughput sequencing of 18S rRNA. The results showed that the NTS had 9.1-12.2% higher percentage of soil macro-aggregates, resulting in 9.8% increase in mean weight diameter and 10.0% increase in bulk soil organic carbon content as compared with CT treatment. In addition, the NTS treatment had significantly higher percentages of Septoglomus and Glomus than the CT treatment. We also found some significant differences in the fungal communities of the soils of the two treatments. There was a strong positive relationship between bulk soil organic carbon and the percentages of Septoglomus and Glomus. Our results suggested that the NTS treatment had a protective effect on AM fungal community structures, which might play a key role in the development of agricultural sustainability in the Loess Plateau of China.

The twin-arginine translocation (Tat) system is a specialized secretion pathway required for bacteria to export fully folded proteins through the cytoplasmic membrane. This system is crucial during Salmonella infection of animal hosts. In this study, we show that Salmonella enterica serovar Typhimurium (S. Typhimurium) requires the Tat system to survive and proliferate intracellularly in the social amoeba Dictyostelium discoideum. To achieve this, we developed a new infection assay to assess intracellular bacterial loads in amoeba by direct enumeration of colony forming units (CFU) at different times of infection. Using this assay we observed that a ΔtatABC mutant was internalized in higher numbers than the wild type, and was defective for intracellular survival in the amoeba at all times post infection evaluated. In addition, we assessed the effect of the ΔtatABC mutant in the social development of D. discoideum. In contrast to the wild-type strain, we observed that the mutant was unable to delay the social development of the amoeba at 2 days of co-incubation. This phenotype correlated with defects in intracellular proliferation presented by the ΔtatABC mutant in D. discoideum after 24 h of infection. All phenotypes described for the mutant were reverted by the presence of a plasmid carrying tatABC genes, indicating that abrogation of Tat system attenuates S. Typhimurium in this model organism. Overall, our results indicate that the Tat system is crucial for S. Typhimurium to survive and proliferate intracellularly in D. discoideum and for virulence in this host. To the best of our knowledge, this is the first report on the relevance of the Tat system in the interaction of any bacterial pathogen with the social amoeba D. discoideum.

The molecular mechanisms of microbial adaptation to repeated anoxic-aerobic cycling were investigated by integrating whole community gene expression (metatranscriptomics) and physiological responses, including the production of nitric (NO) and nitrous (N2O) oxides. Anoxic-aerobic cycling was imposed for 17 days in a lab-scale full-nitrification mixed culture system. Prior to cycling, NO and N2O levels were sustained at 0.097 ± 0.006 and 0.054 ± 0.019 ppmv, respectively. Once the anoxic-aerobic cycling was initiated, peak emissions were highest on the first day (9.8 and 1.3 ppmv, respectively). By the end of day 17, NO production returned to pre-cycling levels (a peak of 0.12 ± 0.007 ppmv), while N2O production reached a new baseline (a peak of 0.32 ± 0.05 ppmv), one order of magnitude higher than steady-state conditions. Concurrently, post-cycling transcription of norBQ and nosZ returned to pre-cycling levels after an initial 5.7- and 9.5-fold increase, while nirK remained significantly expressed (1.6-fold) for the duration of and after cycling conditions. The imbalance in nirK and nosZ mRNA abundance coupled with continuous conversion of NO to N2O might explain the elevated post-cycling baseline for N2O. Metatranscriptomic investigation notably indicated possible NO production by NOB under anoxic-aerobic cycling through a significant increase in nirK expression. Opposing effects on AOB (down-regulation) and NOB (up-regulation) CO2 fixation were observed, suggesting that nitrifying bacteria are differently impacted by anoxic-aerobic cycling. Genes encoding the terminal oxidase of the electron transport chain (ccoNP, coxBC) were the most significantly transcribed, highlighting a hitherto unexplored pathway to manage high electron fluxes resulting from increased ammonia oxidation rates, and leading to overall, increased NO and N2O production. In sum, this study identified underlying metabolic processes and mechanisms contributing to NO and N2O production through a systems-level interrogation, which revealed the differential ability of specific microbial groups to adapt to sustained operational conditions in engineered biological nitrogen removal processes.

Heterologous immunity of virus-specific T cells poses a potential barrier to transplantation tolerance. Cross-reactivity to HLA-A and -B molecules has broadly been described, whereas responses to allo-HLA-C have remained ill defined. In contrast to the transplant setting, HLA-C is the only polymorphic HLA molecule expressed by extravillous trophoblasts at the maternal-fetal interface during pregnancy. Uncontrolled placental viral infections, accompanied by a pro-inflammatory milieu, can alter the activation status and stability of effector T cells. Potential cross-reactivity of maternal decidual virus-specific T cells to fetal allo-HLA-C may thereby have detrimental consequences for the success of pregnancy. To explore the presence of cross-reactivity to HLA-C and the other non-classical HLA antigens expressed by trophoblasts, HLA-A and -B-restricted CD8+ T cells specific for Epstein-Barr virus, Cytomegalovirus, Varicella-Zoster virus, and Influenza virus were tested against target cells expressing HLA-C, -E, and -G molecules. An HLA-B*08:01-restricted EBV-specific T cell clone displayed cross-reactivity against HLA-C*01:02. Furthermore, cross-reactivity of HLA-C-restricted virus-specific CD8+ T cells was observed for HCMV HLA-C*06:02/TRA CD8+ T cell lines and clones against HLA-C*03:02. Collectively, these results demonstrate that cross-reactivity against HLA-C can occur and thereby may affect pregnancy outcome.

N-glycosylation is an essential protein modification that plays roles in many diverse biological processes including protein folding, quality control and protein interactions. Despite recent advances in characterization of the N-glycosylation and N-glycan processing machinery our understanding of N-glycosylation related processes in plant development is limited. In Arabidopsis thaliana, failure of mannose trimming from oligomannosidic N-glycans in the endoplasmic reticulum (ER) and cis/medial-Golgi leads to a defect in root development in the mns123 triple mutant. Here, we show that the severe root phenotype of mns123 is restored in asparagine-linked glycosylation (ALG)-deficient plants with distinct defects in the biosynthesis of the lipid-linked oligosaccharide precursor. The root growth of these ALG-deficient plants is not affected by the α-mannosidase inhibitor kifunensine. Genetic evidence shows that the defect is uncoupled from the glycan-dependent ER-associated degradation (ERAD) pathway that removes misfolded glycoproteins with oligomannosidic N-glycans from the ER. Restoration of mannose trimming using a trans-Golgi targeted α-mannosidase suppresses the defect of mns123 roots. These data suggest that processing of terminal mannose residues from oligomannosidic N-glycans is important for an unknown late-Golgi or post-Golgi process that is implicated in proper root formation.

Multiparent advanced generation intercross (MAGIC) populations were recently developed to allow the high-resolution mapping of quantitative traits. We present a genetic linkage map of an elite but highly diverse eight-founder MAGIC population in common wheat (Triticum aestivum L.). Our MAGIC population is composed of 394 F6:8 recombinant inbred lines lacking significant signatures of population structure. The linkage map included 5435 SNP markers distributed over 2804 loci and spanning 5230 cM. The analysis of population parameters, including genetic structure, kinship, founder probabilities, and linkage disequilibrium and congruency to other maps indicated appropriate construction of both the population and the genetic map. It was shown that eight-founder MAGIC populations exhibit a greater number of loci and higher recombination rates, especially in the pericentromeric regions, compared to four-founder MAGIC, and biparental populations. In addition, our greatly simplified eight-parental MAGIC mating design with an additional eight-way intercross step was found to be equivalent to a MAGIC design with all 210 possible four-way crosses regarding the levels of missing founder assignments and the number of recombination events. Furthermore, the MAGIC population captured 71.7% of the allelic diversity available in the German wheat breeding gene pool. As a proof of principle, we demonstrated the application of the resource for quantitative trait loci mapping analyzing seedling resistance to powdery mildew. As wheat is a crop with many breeding objectives, this resource will allow scientists and breeders to carry out genetic studies for a wide range of breeder-relevant parameters in a single genetic background and reveal possible interactions between traits of economic importance.

Changes in Norwegian law and health policy require all health professionals to help safeguard the provision of information and follow-up for the children of parents with mental or physical illness, or substance abuse problems, to decrease their risk of psychosocial problems. There is a lack of knowledge on how the national changes have been received by hospital-based health professionals, and if they have led to an increase in family focused practice.

i-Scan is a digital image enhancement technology, reported to improve diagnostic performance during endoscopy. Previous studies have investigated the accuracy of i-scan for distinguishing between neoplastic and non-neoplastic colonic polyps and suggested diagnostic accuracy close to that required for use in routine clinical practice. The aim of this study was to perform a meta-analysis of the available literature investigating diagnostic accuracy for i-scan optical diagnosis when made in real time for colorectal polyps <10 mm in size.

The scientific world is facing a constant problem of ghostwriters. These ghostwriters are often attached to the medical publishing houses and are involved in writing an article for a pharmaceutical company which may, in turn, use the name of an established or a famous scientist as an author to the article. Often, such articles are published in well-known journals and are circulating widely. Many a time the adverse effects are overlooked in such papers. This will result in a corrupt practice of prescribing a drug which is not scientifically useful or may have life-threatening side effects. In this present article, the authors discuss this evil practice of ghostwriting in the context of the present day scientific publishing.

The current study examined the effect of single nucleotide (SNPs) polymorphisms in the ADIPOQ gene (I146T and G276T) on body mass index (BMI) of young adult women. The women were divided into underweight, normal, overweight and obese according to BMI. The circulating levels of adiponectin were measured using commercially available ELISA kits. Genetic polymorphisms were genotyped using the PCR-RFLP method. G276T and I164T SNPs are common in the examined population as the frequency of G allele of 276 SNP was 54.8% and for the T allele of 164 SNP it was 41.7%. Circulating adiponectin levels were related to BMI and were lowest in the obese versus overweight, normal weight and underweight groups (p<0.01). However, ADIPOQ gene SNPs (I146T and G276T) showed no association with BMI groups. In conclusion, the results may suggest that adiponectin level, but not ADIPOQ gene SNPs, is a good indicator to BMI in young adult women.

The microRNA (miRNA) cargo contained in plasma extracellular vesicles (EVs) offers a relatively little explored source of biomarkers for brain diseases that can be obtained noninvasively. Methods to isolate EVs from plasma, however, are still being developed. For EV isolation, it is important to ensure the removal of vesicle-free miRNAs, which account for approximately two-thirds of plasma miRNAs. Membrane particle precipitation-based EV isolation is an appealing method because of the simple protocol and high yield. Here, we evaluated the performance of a precipitation-based method to obtain enriched EV-specific miRNAs from a small volume of rat plasma. We performed size-exclusion chromatography (SEC) on precipitation-isolated EV pellets and whole plasma. The SEC fractions were analysed using Nanoparticle Tracking Analysis (NTA), protein and miRNA concentration assays, and droplet digital polymerase chain reaction for four miRNAs (miR-142-3p, miR-124-3p, miR-23a, miR-122). Precipitation-isolated EVs and selected SEC fractions from the plasma were also analysed with transmission electron microscopy (TEM). Precipitation-based EV isolation co-precipitated 9% to 15% of plasma proteins and 21% to 99% of vesicle-free miRNAs, depending on the individual miRNAs. In addition, the amount of miR-142-3p, found mainly in EV fractions, was decreased in the EV fractions, indicating that part of it was lost during precipitation-based isolation. Western blot and TEM revealed both protein and lipoprotein contamination in the precipitation-isolated EV-pellets. Our findings indicate that a precipitation-based method is not sufficient for purifying plasma EV-contained miRNA cargo. The particle number measured by NTA is high, but this is mostly due to the contaminating lipoproteins. Although a part of the vesicle-free miRNA is removed, vesicle-free miRNA still dominates in plasma EV pellets isolated by the precipitation-based method.

Influenza antiviral resistance has been shown to occur in many countries and is commonly found in influenza A(H1N1)pdm09 and A(H3N2). In this study, we monitored and investigated the neuraminidase inhibitor resistance of influenza A(H1N1)pdm09 viruses through the influenza surveillance system in Indonesia.

Our body experience is organized at twofold levels: perceptual and cognitive-emotional. These higher-order processes are clearly different from the primary sensory processing of somatic stimuli (somatosensation). However, most of the available studies have mainly investigated the mechanisms of somatosensation. Moreover, disturbances of our body experience have been documented in some pathological conditions of interest for rehabilitative interventions, but their clinical role and relevance is yet to be clarified. Because in this field we have limited knowledge on perceptual and cognitive body experience, there emerges a need to better clarify this matter. The aim of the present scoping review is to systematically map this topic and to examine the magnitude and the nature of the available evidences.

Physicians who perform colonoscopy should consider appendicitis in the differential diagnosis of post procedure abdominal pain. Diagnostic laparoscopy is a safe adjunct for evaluation in patients with suspected perforation after colonoscopy. It is important that all physicians be aware of this complication to ensure prompt diagnosis and intervention.

The Diet Satisfaction Questionnaire was developed to fill the need for a validated measure to evaluate satisfaction with weight-management diets. This paper further develops the questionnaire, examining the factor structure of the original questionnaire, cross-validating a revised version in a second sample and relating diet satisfaction to weight loss during a 1-year trial.

Obesity is twice as common in people with schizophrenia as the general population and associated with significantly worsened psychiatric and physical health. Despite National Institute for Health and Care Excellence guidelines for the management of psychosis recommending that mental health services offer lifestyle programmes to people with schizophrenia to improve physical health, this is not currently occurring. The aim of the STEPWISE research programme was to develop a lifestyle intervention addressing obesity and preventing weight gain in people with schizophrenia, schizoaffective disorder, or first episode psychosis taking antipsychotic medication, through an approach and fundamental principles drawn from existing diabetes and diabetes prevention interventions. This paper describes the often under-reported process of developing such an intervention from first principles.

With the expansion of the elderly population, age-related osteoporosis and the resulting bone loss have become a significant health and socioeconomic issue. In Triple Energizer (TE)/San Jiao (SJ)/mesenchymal tissue system, mesenchymal stem cell (MSC) senescence, and impaired osteogenesis are thought to contribute to age-related diseases such as osteoporosis. Therefore, comprehending the molecular mechanisms underlying MSC senescence and osteogenesis is essential to improve the treatment of bone metabolic diseases. With the increasing role of miRNAs in MSC aging and osteogenic differentiation, we need to understand further how miRNAs participate in relevant mechanisms. In this study, we observed that the expression of miR-1292 was augmented during cellular senescence and lessened with osteogenesis in human adipose-derived mesenchymal stem cells (hADSCs). miR-1292 expression was positively correlated with senescence markers and negatively associated with bone formation markers in clinical bone samples. Overexpression of miR-1292 notably accelerated hADSC senescence and restrained osteogenesis, whereas its knockdown decreased senescence and enhanced osteogenic differentiation. Furthermore, miR-1292 upregulation inhibited ectopic bone formation in vivo. Mechanistically, FZD4 was identified as a potential target of miR-1292. Downregulation of FZD4 phenocopied the effect of miR-1292 overexpression on hADSC senescence and osteogenic differentiation. Moreover, the impact of miR-1292 suppression on senescence and osteogenesis were reversed by the FZD4 knockdown. Pathway analysis revealed that miR-1292 regulates hADSC senescence and osteogenesis through the Wnt/β-catenin signaling pathway. Thus, TE/SJ/mesenchymal tissue system is the largest organ composed of various functional cells derived from mesoderm, responsible for maintaining homeostasis and regulating cell senescence. miR-1292 might serve as a novel therapeutic target for the prevention and treatment of osteoporosis or other diseases related to bone metabolism and aging.