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Abstract Rifaximin is FDA-approved for treatment of irritable bowel syndrome with diarrhea (IBS-D), but poor solubility may limit its efficacy against microbes in the mucus layer, e.g. Escherichia coli. Here we evaluate adding the mucolytic N-acetylcysteine (NAC) to improve rifaximin efficacy. In a resazurin checkerboard assay, combining rifaximin with NAC had significant synergistic effects in reducing E. coli levels. The optimal rifaximin + NAC combination was then tested in a validated rat model of IBS-D (induced by cytolethal distending toxin [CdtB] inoculation). Rats were inoculated with vehicle and treated with placebo (Control-PBS) or rifaximin + NAC (Control-Rif + NAC, safety), or inoculated with CdtB and treated with placebo (CdtB-PBS), rifaximin (CdtB-Rifaximin), or rifaximin + NAC (CdtB-Rif + NAC) for 10 days. CdtB-inoculated rats (CdtB-PBS) developed wide variability in stool consistency (P = 0.0014) vs. controls (Control-PBS). Stool variability normalized in rats treated with rifaximin + NAC (CdtB-Rif + NAC) but not rifaximin alone (CdtB-Rifaximin). Small bowel bacterial levels were elevated in CdtB-PBS rats but normalized in CdtB-Rif + NAC but not CdtB-Rifaximin rats. E. coli and Desulfovibrio spp levels (each associated with different IBS-D microtypes) were also elevated in CdtB-inoculated (CdtB-PBS) but normalized in CdtB-Rif + NAC rats. Cytokine levels normalized only in CdtB-Rif + NAC rats, in a manner predicted to be associated with reduced diarrhea driven by reduced E. coli. These findings suggest that combining rifaximin with NAC may improve the percentage of IBS-D patients responding to treatment.

ABSTRACTDiarrhea-predominant irritable bowel syndrome (IBS-D), associated with increased intestinal permeability, inflammation, and small intestinal bacterial overgrowth, can be triggered by acute gastroenteritis. Cytolethal distending toxin B (CdtB) is produced by gastroenteritis-causing pathogens and may underlie IBS-D development, through molecular mimicry with vinculin. Here, we examine the effects of exposure to CdtB alone on gut microbiome composition, host intestinal gene expression, and IBS-D-like phenotypes in a rat model. CdtB-inoculated rats exhibited increased anti-CdtB levels, which correlated with increased stool wet weights, pro-inflammatory cytokines (TNFα, IL2) and predicted microbial metabolic pathways including inflammatory responses, TNF responses, and diarrhea. Three distinct ileal microbiome profiles (microtypes) were identified in CdtB-inoculated rats. The first microtype (most like controls) had altered relative abundance (RA) of genera Bifidobacterium, Lactococcus, and Rothia. The second had lower microbial diversity, higher Escherichia-Shigella RA, higher absolute E. coli abundance, and altered host ileal tissue expression of immune-response and TNF-response genes compared to controls. The third microtype had higher microbial diversity, higher RA of hydrogen sulfide (H2S)-producer Desulfovibrio, and increased expression of H2S-associated pain/serotonin response genes. All CdtB-inoculated rats exhibited decreased ileal expression of cell junction component mRNAs, including vinculin-associated proteins. Significantly, cluster-specific microRNA-mRNA interactions controlling intestinal permeability, visceral hypersensitivity/pain, and gastrointestinal motility genes, including several previously associated with IBS were seen. These findings demonstrate that exposure to CdtB toxin alone results in IBS-like phenotypes including inflammation and diarrhea-like stool, decreased expression of intestinal barrier components, and altered ileal microtypes that influenced changes in microRNA-modulated gene expression and predicted metabolic pathways consistent with specific IBS-D symptoms.

Egg production is a significant source of animal protein for human consumption. Feed costs significantly impact the profitability of egg production, representing more than 70% of the variable costs. This study evaluated the effect of dietary intakes of three essential amino acids (EAAs) on the egg cost for H&N Brown second-cycle laying hens. The hens were fed for 20 weeks with 23 diets that varied in their lysine, methionine + cystine, and threonine contents. These amino acids were derived from both dietary and synthetic sources. Zootechnical results were used to calculate the feed cost per kilogram of egg (FCK), considering the cost of raw materials and the diet composition. Multivariate polynomial models and artificial neural networks (ANNs) were validated to predict FCK as a function of the EAAs and time. The EAA intakes that minimize FCK over time were optimized using the best model, a cascade-forward ANN with a softmax transfer function. The optimal scenario for FCK (0.873 USD/kg egg) at 20 weeks was achieved at 943.7 mg lysine/hen-day, 858.3 mg methionine + cystine/hen-day, and 876.8 mg threonine/hen-day. ANNs could be a valuable tool for predicting the egg cost of laying hens based on the nutritional requirements. This could help improve economic efficiency and reduce the feed costs in poultry companies.

In a global context, where the construction industry is a major source of CO2 emissions and resource use, is dependent on concrete and its risks, and lags behind in digitalization, a clear need arises to direct architecture towards more practical, efficient, and sustainable practices. This research introduces an alternative technique for building timber space structures, aiming to expand its applications in areas with limited access to advanced technologies such as CNCs with more than five axes and industrial robotic arms. This involves reconfiguring economic and ecological constraints to maximize the structural and architectural advantages of these systems. The method develops a parametric tool that integrates computational design and manufacturing based on two-axis laser cutting for shells with segmented hexagonal plywood plates. It uses a modified ‘half-lap joint’ mechanical joint, also made of plywood and without additional fasteners, ensuring a precise and robust connection. The results demonstrate the compatibility of the geometry with two-axis CNC machines, which simplifies manufacturing and reduces the cuts required, thus increasing economic efficiency. The prototype, with a span of 1.5 m and composed of 63 plywood panels and 163 connectors, each 6 mm thick, supported a point load of 0.8 kN with a maximum displacement of 5 mm, weighing 15.1 kg. Assembly and disassembly, carried out by two students, took 5 h and 1.45 h, respectively, highlighting the practicality and accessibility of the method. In conclusion, the technique for building timber shells based on two-axis CNC is feasible and effective, proven by practical experimentation and finite element analysis.

Summary: Studies using stool samples suggest that non-sugar sweetener (NSS) consumption affects gut microbiome composition. However, stool does not represent the entire gut. We analyzed the duodenal luminal microbiome in subjects consuming non-aspartame non-sugar sweeteners (NANS, N = 35), aspartame only (ASP, N = 9), and controls (CON, N = 55) and the stool microbiome in a subset (N = 40). Duodenal alpha diversity was decreased in NANS vs. CON. Duodenal relative abundance (RA) of Escherichia, Klebsiella, and Salmonella (all phylum Proteobacteria) was lower in both NANS and ASP vs. CON, whereas stool RA of Escherichia, Klebsiella, and Salmonella was increased in both NANS and ASP vs. CON. Predicted duodenal microbial metabolic pathways altered in NANS vs. CON included polysaccharides biosynthesis and D-galactose degradation, whereas cylindrospermopsin biosynthesis was significantly enriched in ASP vs. CON. These findings suggest that consuming non-sugar sweeteners may significantly alter microbiome composition and function in the metabolically active small bowel, with different alterations seen in stool.

The purpose of this research is to propose an architectural design for an Interpretation Center aimed at the revaluation of the flora and fauna of Cusco, Peru, in 2023. Due to the lack of awareness spaces, deforestation, water pollution, and the planting of introduced species in the area, there is a loss of endemic flora and fauna. The methodologies used in this study included climatic analysis of the location, characterization of the flora and fauna, and the application of bioclimatic design strategies supported by the use of software (AutoCAD, Revit, and 3D Sun-path). Additionally, urban design strategies were employed based on the Inca worldview and the Sustainable Development Goals. The feasibility of implementing this urban and architectural proposal was compared with other projects such as the Sydney Water Restoration Project and the Ataria Nature Interpretation Center. As a result, an ecological network was designed for the preservation of endemic species. The project also includes green spaces, which account for 51% of the total area. These green spaces will facilitate carbon dioxide absorption and contribute to the revaluation of the Huantanay River and the creation of microclimates in the area.

ABSTRACT An experiment was conducted to evaluate the effect of the intake of a mixture of fish and sacha inchi oils (iOM), organic selenium (iSe), and organic chromium (iCr) on egg production (EP) and feed conversion ratio (FCR) of Isa Brown second-cycle laying hens (SCLH) for 16 weeks (91-106 weeks old). Egg production and FCR were evaluated using multivariate models that included conventional equations and artificial neural networks (ANN) to study multiple nutritional interactions as alternatives to univariate dose-response models. Based on the best models, iOM, iSe, and iCr levels were optimized, and a global sensitivity analysis was implemented to quantify their influence on EP and FCR. The modified logistic model was selected as the best strategy to represent EP. In the case of FCR, an ANN model with a feed-forward architecture and softmax transfer function was selected as the best alternative. One of the scenarios to simultaneously optimize EP (89.1%) and FCR (1.94 kg feed/kg egg) at 16 weeks of production was established with 3.3 g/hen·day of iOM, 0.132 mg/ hen·day of iSe, and 0.176 mg/hen·day of iCr. However, optimization considering only FCR results in much lower optimal iCr levels (between 0.083 and 0.105 mg/hen·day) with a slight decrease in EP (87.9%). The global sensitivity analysis showed that iSe is an essential factor associated with the increase in EP, and iCr is the most influential factor for the decrease in FCR. When both criteria were taken into account simultaneously from a desirability function, iSe was the most critical factor.

Gut microbiome composition is different in males and females, but sex is rarely considered when prescribing antibiotics, and sex-based differences in gut microbiome recovery following antibiotic treatment are poorly understood. Here, we compared the effects of broad-spectrum antibiotics on both the stool and small bowel microbiomes in male and female rats. Adult male and female Sprague Dawley rats were exposed to a multi-drug antibiotic cocktail for 8 days, or remained unexposed as controls. Following cessation of antibiotics, rats were monitored for an additional 13-day recovery period prior to euthanasia. Baseline stool microbiome composition was similar in males and females. By antibiotic exposure day 8 (AbxD8), exposed male rats exhibited greater loss of stool microbial diversity compared to exposed females, and the relative abundance (RA) of numerous taxa were significantly different in exposed males vs. exposed females. Specifically, RA of phylum Proteobacteria and genera Lactobacillus, Sutterella, Akkermansia, and Serratia were higher in exposed males vs. exposed females, whereas RA of phyla Firmicutes and Actinobacteria and genera Turicibacter and Enterococcus were lower. By 13 days post antibiotics cessation (PAbxD13), the stool RA of these and other taxa remained significantly different from baseline, and also remained significantly different between exposed males and exposed females. RA of phyla Firmicutes and Actinobacteria and genus Enterococcus remained lower in exposed males vs. exposed females, and genus Sutterella remained higher. However, RA of phylum Proteobacteria and genus Akkermansia were now also lower in exposed males vs. females, whereas RA of phylum Bacteroidetes and genus Turicibacter were now higher in exposed males. Further, the small bowel microbiome of exposed rats on PAbxD13 was also significantly different from unexposed controls, with higher RA of Firmicutes, Turicibacter and Parabacteroides in exposed males vs. females, and lower RA of Bacteroidetes, Proteobacteria, Actinobacteria, Oscillospira, Sutterella, and Akkermansia in exposed males vs. females. These findings indicate that broad-spectrum antibiotics have significant and sex-specific effects on gut microbial populations in both stool and the small bowel, and that the recovery of gut microbial populations following exposure to broad-spectrum antibiotics also differs between sexes. These findings may have clinical implications for the way antibiotics are prescribed.

ABSTRACT An experiment with 23 diets was performed to evaluate the effect of digestible lysine (Lys), digestible methionine + cysteine (Met+Cys), and digestible threonine (Thr) on egg production of H&N Brown second-cycle laying hens (SCLH) for 20 weeks (92-111 weeks of age) in cages under environmental conditions. Body weight (BW), feed intake (FI), feed conversion ratio (FCR), egg weight (EW), number of hen-housed eggs, and livability were also evaluated during the experiment. Diets were formulated from a central composite design that combined five levels of Lys, Met+Cys, and Thr ranging from 727 to 1159, 662 to 1055, and 552 to 882 mg/kg, respectively. Egg production (EP) data were evaluated through three different modeling strategies: egg production models, multivariate polynomial models, and artificial neural networks (ANN). A cascade-forward neural network with log-sigmoid transfer function was selected as the best model according to goodness-of-fit statistics in both identification and validation data. One of the best scenarios for EP of H&N Brown SCLH under specific outdoor conditions was established at Lys, Met+Cys, and Thr levels of 1138, 1031, and 717 mg/hen·day, respectively. The ANN model may be an appropriate tool to study and predict EP of H&N Brown SCLH based on the combination of three different levels of essential digestible amino acids. The strategies included in this work may contribute to improving poultry performance based on modeling techniques to study other production parameters in terms of different nutritional requirements and productive conditions.

Abstract Background The human small intestine plays a central role in the processes of digestion and nutrient absorption. However, characterizations of the human gut microbiome have largely relied on stool samples, and the associated methodologies are ill-suited for the viscosity and low microbial biomass of small intestine samples. As part of the REIMAGINE study to examine the specific roles of the small bowel microbiome in human health and disease, this study aimed to develop and validate methodologies to optimize microbial analysis of the small intestine. Results Subjects undergoing esophagogastroduodenoscopy without colon preparation for standard of care were prospectively recruited, and ~ 2 ml samples of luminal fluid were obtained from the duodenum using a custom sterile aspiration catheter. Samples of duodenal aspirates were either untreated (DA-U, N = 127) or pretreated with dithiothreitol (DA-DTT, N = 101), then cultured on MacConkey agar for quantitation of aerobic gram-negative bacteria, typically from the class Gammaproteobacteria, and on blood agar for quantitation of anaerobic microorganisms. DA-DTT exhibited 2.86-fold greater anaerobic bacterial counts compared to DA-U (P = 0.0101), but were not statistically different on MacConkey agar. DNA isolation from DA-U (N = 112) and DA-DTT (N = 43) samples and library preparation for 16S rRNA gene sequencing were also performed using modified protocols. DA-DTT samples exhibited 3.81-fold higher DNA concentrations (P = 0.0014) and 4.18-fold higher 16S library concentrations (P

Objectives To compare the appendiceal microbiomes and examine the prevalence of Campylobacter species in the appendices of adult subjects with confirmed acute non-perforated appendicitis and controls with healthy appendices.Design Archived samples of formalin-fixed paraffin-embedded appendiceal tissues were obtained from 50 consecutive female subjects who underwent appendectomy for acute, non-perforated appendicitis, and 35 consecutive female controls who underwent incidental appendectomy during gynaecological surgery.Results 16S rRNA gene sequencing revealed that the relative abundances (RAs) of the major phyla in appendiceal tissues (Firmicutes, Proteobacteria, Bacteroidetes, and Actinobacteria) were similar in both groups. Beta diversity was significantly different due to differences in Bacteroidetes and Proteobacteria (p0.05) were increased in acute appendicitis samples. RAs of unknown genera from families Burkholderiaceae and Enterobacteriaceae were decreased in appendicitis samples, and 14 genera were increased, including Neisseria, Acinetobacter and Campylobacter. Quantitative PCR revealed that levels of Campylobacter jejuni DNA, but not other Campylobacter species or pathogens tested, were significantly higher in appendicitis samples than in controls (p=0.013). Using a cut-off of 0.31 pg/µL, 40% of appendicitis cases and 6% of controls were positive for C. jejuni, indicating specificity of 93.7% (95% Cl 79.2 to 99.2), sensitivity of 40.9% (95% Cl 24.7 to 54.5), and OR of 10.38 (Fisher’s p value=0.0006, 95% Cl 2.3 to 47.4).Conclusions Our findings indicate that Campylobacter jejuni may be a significant cause of acute appendicitis. This supports earlier studies and suggests that targeted antibiotic therapies could be an alternative treatment for a subset of non-complicated acute appendicitis cases.

Antimicrobial-resistant and novel pathogens continue to emerge, outpacing efforts to contain and treat them. Therefore, there is a crucial need for safe and effective therapies. Ultraviolet-A (UVA) phototherapy is FDA-approved for several dermatological diseases but not for internal applications. We investigated UVA effects on human cells in vitro, mouse colonic tissue in vivo, and UVA efficacy against bacteria, yeast, coxsackievirus group B and coronavirus-229E. Several pathogens and virally transfected human cells were exposed to a series of specific UVA exposure regimens. HeLa, alveolar and primary human tracheal epithelial cell viability was assessed after UVA exposure, and 8-Oxo-2'-deoxyguanosine was measured as an oxidative DNA damage marker. Furthermore, wild-type mice were exposed to intracolonic UVA as an in vivo model to assess safety of internal UVA exposure. Controlled UVA exposure yielded significant reductions in Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Clostridioides difficile, Streptococcus pyogenes, Staphylococcus epidermidis, Proteus mirabilis and Candida albicans. UVA-treated coxsackievirus-transfected HeLa cells exhibited significantly increased cell survival compared to controls. UVA-treated coronavirus-229E-transfected tracheal cells exhibited significant coronavirus spike protein reduction, increased mitochondrial antiviral-signaling protein and decreased coronavirus-229E-induced cell death. Specific controlled UVA exposure had no significant effect on growth or 8-Oxo-2'-deoxyguanosine levels in three types of human cells. Single or repeated in vivo intraluminal UVA exposure produced no discernible endoscopic, histologic or dysplastic changes in mice. These findings suggest that, under specific conditions, UVA reduces various pathogens including coronavirus-229E, and may provide a safe and effective treatment for infectious diseases of internal viscera. Clinical studies are warranted to further elucidate the safety and efficacy of UVA in humans.

Small intestinal bacterial overgrowth (SIBO) is highly prevalent and is associated with numerous gastrointestinal disorders, but the microbes involved remain poorly defined. Moreover, existing studies of microbiome alterations in SIBO have utilized stool samples, which are not representative of the entire gastrointestinal tract. Therefore, we aimed to determine and compare the duodenal microbiome composition in SIBO and non-SIBO subjects, using duodenal aspirates from subjects undergoing standard-of-care esophagogastroduodenoscopy without colon preparation. Using the recently-redefined cutoff for SIBO of >103 colony forming units per milliliter (CFU/mL), 42 SIBO and 98 non-SIBO subjects were identified. Duodenal samples from SIBO subjects had 4x103-fold higher counts than non-SIBO subjects when plated on MacConkey agar (P103 CFU/mL cutoff for the definition of SIBO, and also reveal specific overgrowth of Proteobacteria in SIBO vs. non-SIBO subjects, coupled with an altered Proteobacterial profile that correlates with symptom severity. Future research may elucidate host-microbiome interactions underlying these symptoms in SIBO patients.

Autonomous vehicles are expected to display human-like behavior, at least to the extent that their decisions can be intuitively understood by other road users. If this is not the case, the coexistence of manual and autonomous vehicles in a mixed environment might affect road user interactions negatively and might jeopardize road safety. To this end, it is highly important to design algorithms that are capable of analyzing human decision-making processes and of reproducing them. In this context, lane-change maneuvers have been studied extensively. However, not all potential scenarios have been considered, since most works have focused on highway rather than urban scenarios. We contribute to the field of research by investigating a particular urban traffic scenario in which an autonomous vehicle needs to determine the level of cooperation of the vehicles in the adjacent lane in order to proceed with a lane change. To this end, we present a game theory-based decision-making model for lane changing in congested urban intersections. The model takes as input driving-related parameters related to vehicles in the intersection before they come to a complete stop. We validated the model by relying on the Co-AutoSim simulator. We compared the prediction model outcomes with actual participant decisions, i.e., whether they allowed the autonomous vehicle to drive in front of them. The results are promising, with the prediction accuracy being 100% in all of the cases in which the participants allowed the lane change and 83.3% in the other cases. The false predictions were due to delays in resuming driving after the traffic light turned green.

Diarrhea-predominant irritable bowel syndrome (IBS) is diagnosed through clinical criteria after excluding "organic" conditions, and can be precipitated by acute gastroenteritis. Cytolethal distending toxin B (CdtB) is produced by bacteria that cause acute gastroenteritis, and a post-infectious animal model demonstrates that host antibodies to CdtB cross-react with vinculin in the host gut, producing an IBS-like phenotype. Therefore, we assessed circulating anti-CdtB and anti-vinculin antibodies as biomarkers for D-IBS in human subjects. Subjects with D-IBS based on Rome criteria (n=2375) were recruited from a large-scale multicenter clinical trial for D-IBS (TARGET 3). Subjects with inflammatory bowel disease (IBD) (n=142), subjects with celiac disease (n=121), and healthy controls (n=43) were obtained for comparison. Subjects with IBD and celiac disease were recruited based on the presence of intestinal complaints and histologic confirmation of chronic inflammatory changes in the colon or small intestine. Subjects with celiac disease were also required to have an elevated tTG and biopsy. All subjects were aged between 18 and 65 years. Plasma levels of anti-CdtB and anti-vinculin antibodies were determined by ELISA, and compared between groups. Anti-CdtB titers were significantly higher in D-IBS subjects compared to IBD, healthy controls and celiac disease (P

El propósito de esta investigación consistió en evaluar los efectos de dos colutorios orales (un colutorio con fluoruro de sodio al 0,05% y un colutorio de agua con sal al 5%) sobre la placa bacteriana y el pH salival en una población de soldados del ejercito. La muestra estuvo conformada por tres grupos: un grupo uso un colutorio de fluoruro de sodio al 0,05%, otro uso un colutorio de agua con sal y un tercer grupo control utiltzó agua destilada durante 40 días. Las características de la población permitió controlar importantes variables intervinientes tales como la dieta, higiene bucal de los soldados, momentos de higiene y cumplimiento del protocolo señalado. Se usó un pHmetro calibrado y el índice simplificado de Green y Vermillon, para medir el pH salival y el índice de placa bacteriana respectivamente. Se utilizó el análisis de varianza, la prueba F y la prueba T de student para muestras apareadas y establecer diferencias. Los resultados del trabajo de investigación indicaron que tanto el colutorio con fluoruro de sodio al 0,05% como e1 colutorio con agua con sal al 5% se comportan de manera similar sobre la placa bacteriana, disminuyéndola al cabo de 40 días; mientras que ninguno de los colutorios fue capaz de alterar el pH salival de forma significativa.