Role of rs2282679 in 25-hydroxyvitamin D levels and insulin resistance after a sleeve gastrectomy.

BACKGROUND & AIMS: Some studies have reported links between 25-hydroxyvitamin D levels and the presence of obesity and some genetic variants. The aim of our design was to evaluate the effects of rs2282679 genetic variant of CG gene on 25-hydroxyvitamin D levels, weight loss and metabolic parameters after a robotic sleeve gastrectomy in premenopausal females with obesity. METHODS: 76 participants were enrolled. 25-hydroxyvitamin D levels, biochemical evaluation and anthropometric parameters were registered before surgery and after 3, 6 and 12 months follow up. Genotype of rs2282679 CG gene was evaluated. RESULTS: The improvements in anthropometric parameters, blood pressure and lipid profile were similar in both genotypes (TT vs TG + GG). Basal insulin levels and HOMA-IR were greater in G allele carriers than non-carriers (Delta: 6.7 ± 1.2 mUI/L; p = 0.01) and (Delta: 1.3 ± 0.1 units; p = 0.02). 25-hydroxyvitamin D levels were lower in G allele carriers than non-carriers (Delta: 8.1 ± 1.1 ng/dl; p = 0.03). The levels of insulin and HOMA-IR remained greater in G allele carriers than non-carriers throughout all the visits. The levels of 25-hydroxyvitamin D remained lower in G allele carriers than non-G allele. The average level of 25-hydroxyvitamin D at 12 months in non-G allele carriers were above 30 ng/dl (36.0 ± 3.1 ng/dl) and the level in G allele carriers were below (24.9 ± 4.9 ng/dl). CONCLUSIONS: rs 2282679 (GC) was related with low 25 hydroxyvitamin D levels and insulin resistance. In addition, the presence of G allele produced a decrease in the improvement of 25-hydroxyvitamin D levels and insulin resistance after weight loss during 12 months.

SOX9 switch links regeneration to fibrosis at the single-cell level in mammalian kidneys.

The steps governing healing with or without fibrosis within the same microenvironment are unclear. After acute kidney injury (AKI), injured proximal tubular epithelial cells activate SOX9 for self-restoration. Using a multimodal approach for a head-to-head comparison of injury-induced SOX9 lineages, we identified a dynamic SOX9 switch in repairing epithelia. Lineages that regenerated epithelia silenced SOX9 and healed without fibrosis (SOX9on-off). By contrast, lineages with unrestored apicobasal polarity maintained SOX9 activity in sustained efforts to regenerate, which were identified as a SOX9on-on Cadherin6pos cell state. These reprogrammed cells generated substantial single-cell WNT activity to provoke a fibroproliferative response in adjacent fibroblasts, driving AKI to chronic kidney disease. Transplanted human kidneys displayed similar SOX9/CDH6/WNT2B responses. Thus, we have uncovered a sensor of epithelial repair status, the activity of which determines regeneration with or without fibrosis.

Breaching septal veins while attempting left bundle branch area pacing. If necessary, septography contrast should be injected in a stepwise approach.

Permanent transseptal left bundle branch area pacing (LBBAP) is a promising technique developed to avoid the detrimental effects of pacing-induced dyssynchrony with right ventricular (RV) pacing, by offering more physiologic activation of the heart. Lesions to tributary veins of the coronary sinus have been increasingly reported, mostly associated with venous fistula or venous septal system infringement. Despite being mostly benign, venous complications may be related to the maneuver of contrast injection through the sheath and failure to follow simple but essential steps.

[Diet as a source of hydrogen sulfide and its effects on health and disease]. / La dieta como fuente de sulfuro de hidrógeno y sus efectos en la salud y la enfermedad.

Introduction: Initially known for its deleterious health effects, hydrogen sulfide (H2S) has recently been recognized as a biologically important gas carrier, like nitric oxide and carbon monoxide. H2S is produced endogenously in mammalian cells by enzymatic and non-enzymatic pathways. When it is produced by the enzymatic pathway, its synthesis is carried out from the amino acid L-cysteine through the transsulfuration pathway. It can also be produced endogenously from exogenous compounds that function as H2S donors as, for example, the naturally occurring organic donors found in some plants. Currently, the role of S2H is well known as brain and cardiac protector, and its research as a therapeutic adjuvant in metabolic diseases such as obesity and type-2 diabetes is becoming increasingly important. The objective of this review is to examine how the contribution of donors and precursors of hydrogen sulfide by the diet impacts health and disease.

Introducción: Conocido originalmente por sus efectos deletéreos en la salud, recientemente se ha reconocido al sulfuro de hidrógeno (H2S) como un gasotransmisor de importancia biológica, al igual que el óxido nítrico y el monóxido de carbono. El H2S puede producirse de forma endógena en las células de mamíferos por dos vías: la vía enzimática y la vía no enzimática. Cuando se produce por la vía enzimática, su síntesis se lleva a cabo a partir de los aminoácidos L-cisteína o metionina mediante transulfuración y transmetilación. También se puede producir el H2S a partir donadores de grupos sulfuro como, por ejemplo, compuestos orgánicos que se encuentran presentes en algunos vegetales. Actualmente es bien conocido el papel del H2S como protector a nivel cerebral y cardiaco, y cada vez adquiere mayor relevancia su estudio como coadyuvante terapéutico en padecimientos metabólicos como la obesidad y la diabetes mellitus de tipo 2. El objetivo de esta revisión es examinar cómo impacta el aporte de donadores y precursores del sulfuro de hidrogeno por la dieta en la salud y la enfermedad.

The significance of N-carbamoylglutamate in ruminant production.

Improving the efficiency and production of grazing ruminants to support food and fiber production, while reducing the environmental footprint and meeting the welfare needs of the animals, is important for sustainable livestock production systems. Development of new technologies that can improve the efficiency of nitrogen (N) utilization in ruminants, and that are effective and safe, has important implications for ruminant livestock production. N-carbomoylglutamate (NCG) is a functional micronutrient that stimulates endogenous synthesis of arginine, which can improve survival, growth, lactation, reproductive performance, and feed efficiency in mammals. There is a growing body of evidence to support the potential of dietary NCG supplementation to improve the productive capacity and N utilization efficiency of ruminants. This review summarizes the current literature on the effects of dietary supplementation with NCG in ruminants and impacts on production and potential to reduce the environmental footprint of farmed ruminant livestock. The current literature highlights the potential for commercial application in ruminant livestock to improve productivity and N utilization efficiency.

Inhibition of ATP synthase reverse activity restores energy homeostasis in mitochondrial pathologies.

The maintenance of cellular function relies on the close regulation of adenosine triphosphate (ATP) synthesis and hydrolysis. ATP hydrolysis by mitochondrial ATP Synthase (CV) is induced by loss of proton motive force and inhibited by the mitochondrial protein ATPase inhibitor (ATPIF1). The extent of CV hydrolytic activity and its impact on cellular energetics remains unknown due to the lack of selective hydrolysis inhibitors of CV. We find that CV hydrolytic activity takes place in coupled intact mitochondria and is increased by respiratory chain defects. We identified (+)-Epicatechin as a selective inhibitor of ATP hydrolysis that binds CV while preventing the binding of ATPIF1. In cells with Complex-III deficiency, we show that inhibition of CV hydrolytic activity by (+)-Epichatechin is sufficient to restore ATP content without restoring respiratory function. Inhibition of CV-ATP hydrolysis in a mouse model of Duchenne Muscular Dystrophy is sufficient to improve muscle force without any increase in mitochondrial content. We conclude that the impact of compromised mitochondrial respiration can be lessened using hydrolysis-selective inhibitors of CV.

Murine-Derived Glioma Organoids and Cell Line Culture Systems.

Research models in cancer have greatly evolved in the last decade, with the advent of several new methods both in vitro and in vivo. While in vivo models remain the gold standard for preclinical studies, these methods present a series of disadvantages such as a high cost and long periods of time to produce results compared with in vitro models. We have previously developed a method named Mosaic Analysis by Dual Recombinase-mediated cassette exchange (MADR) that generates autochthonous gliomas in immunocompetent mice through the transgenesis of personalized driver mutations, which highly mimic the spatial and temporal tumor development of their human counterparts. Due to the control of single-copy expression of transgenes, it allows for comparing the visualization of tumor cells and non-tumor cells. Here we describe a method to generate murine-derived glioma organoids (MGOs) and cell line cultures from these murine models by physical and enzymatic methods for in vitro downstream applications. Tumor cells can be readily distinguished from non-tumor cell populations, in both organoids and monolayer cell cultures, and isolated due to the use of personalized fluorescent reporter transgenes. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Generation of 3D murine-derived glioma organoids Basic Protocol 2: Generation of 2D glioma monolayer cell lines.

Meta-Regression to Develop Predictive Equations for Urinary Nitrogen Excretion of Lactating Dairy Cows.

Dairy cows' urinary nitrogen (N) excretion (UN; g/d) represents a significant environmental concern due to their contribution to nitrate leaching, nitrous oxide (a potent greenhouse gas), and ammonia emissions (contributor to N deposition). The first objective of the current study was to determine the adequacy of existing models to predict UN from total mixed ration (TMR)-fed and fresh forage (FF)-fed cows. Next, we aimed to develop equations to predict UN based on animal factors [milk urea nitrogen (MUN; mg/dL) and body weight (BW, kg)] and to explore how these equations are improved when dietary factors, such as diet type, dry matter intake (DMI), or dietary characteristics [neutral detergent fiber (NDF) and crude protein (CP) content], are considered. A dataset was obtained from 51 published experiments composed of 174 treatment means. The whole dataset was used to evaluate the mean and linear biases of three existing equations including diet type as an interaction term; all models had significant linear and mean biases and two of the three models had poor predictive capabilities as indicated by their large relative prediction error (RPE; root mean square error of prediction as a percent of the observed mean). Next, the complete data set was split into training and test sets, which were used to develop and to evaluate new models, respectively. The first model included MUN and BW, and there was a significant interaction between diet type and the coefficients. This model had the worst 1:1 agreement [Lin's concordance correlation coefficient (CCC) = 0.50] and largest RPE (24.7%). Models that included both animal and dietary factors performed the best, and when included in the model, the effect of diet type was no longer significant (p > 0.10). These models all had very good agreement (CCC ≥ 0.86) and relatively low RPE (≤13.1%). This meta-analysis developed precise and accurate equations to predict UN from dairy cows in both confined and pasture-based systems.

A single FTO gene variant rs9939609 is associated with weight change and insulin resistance improvement in response to a robotic sleeve gastrectomy in individuals with severe obesity.

BACKGROUND: Genetic mechanisms have been involved in the weight response secondary to bariatric surgery. OBJECTIVE: The aim of our study was to evaluate the effects of the rs9939609 genetic variant on weight loss and metabolic parameters after sleeve gastrectomy. SETTING: Tertiary hospital. METHODS: A total of 95 participants were enrolled. Co-morbidities, biochemical evaluation, and anthropometric parameters were registered before and after 3-, 6-, and 12-month follow-up. Genotype of the rs9939609 fat mass and obesity-associated (FTO) gene was evaluated. RESULTS: We grouped the participants into 2 groups: carriers of A allele (TA+AA, 69.5%) and noncarriers of A allele (TT, 30.5%). We detected a statistically significant reduction of blood pressure, biochemical, and anthropometric parameters at 3 times during follow-up. After 6 months, changes of some parameters were greater in non-A allele carriers: weight (-39.6 + 4.0 kg versus -24.6 + 2.8 kg; P = .02), waist circumference (-21.1 + 2.1 cm versus -16.2 + 1.8 cm; P = .04), insulin (-12.3 + .9 mUI/L versus -8.9.1 + .2 mUI/L; P = .02), and homeostasis model assessment of insulin resistance (-3.1 + .1 units versus -2.3 + .1 units; P = .02 ). After 12 months, changes of the aforementioned parameters remained greater in non-A allele carriers. The percentage of participants with diabetes diminished earlier in the non-A allele carriers than A allele carriers at 6-month follow-up. The percentage of participants with diabetes at the end of the study was lower in non-A allele carriers (3.4% versus 12.1%; P = .02). CONCLUSIONS: Our data suggest that non-A allele carriers of the genetic variant (rs9939609) of the FTO gene showed a better improvement of anthropometric and insulin levels in non-A allele carriers after a robotic sleeve gastrectomy. Both improvements are associated with a lower percentage of participants with diabetes at 12 months.

Evaluation of Feed Near-Infrared Reflectance Spectra as Predictors of Methane Emissions from Ruminants.

Feed chemical composition is associated with methane (CH4) formation in the rumen, and thus CH4 yields (Ym; CH4 emitted from per unit of dry matter intake) could be predicted using near-infrared reflectance spectroscopy (NIRS) of feeds fed to ruminants. Two databases of NIRS data were compiled from feeds used in experiments in which CH4 yields had been quantified in respiration chambers. Each record in the databases represented a batch of feed offered to a group of experimental animals and the mean CH4 yield for the group. A near-infrared reflectance spectrum was obtained from each feed, and these spectra were used to generate a predictive equation for Ym. The predictive model generated from brassica crops and pasture fed at a similar feeding level (n = 40 records) explained 53% of the variation in Ym and had a reasonably good agreement (concordance correlation coefficient of 0.77). The predictive ability of the NIRS calibration could be useful for screening purposes, particularly for predicting the potential Ym of multiple feeds or feed samples, rather than measuring Ym in animal experiments at high expenses. It is recommended that the databases for NIRS calibrations are expanded by collecting feed information from future experiments in which methane emissions are measured, using alternative algorithms and combining other techniques, such as terahertz time-domain spectroscopy.

A Review: Plant Carbohydrate Types-The Potential Impact on Ruminant Methane Emissions.

Carbohydrates are the major component of most ruminant feeds. The digestion of carbohydrates in the rumen provides energy to the ruminants but also contributes to enteric methane (CH4) emissions. Fresh forage is the main feed for grazing ruminants in temperate regions. Therefore, this review explored how dietary carbohydrate type and digestion affect ruminant CH4 emissions, with a focus on fresh forage grown in temperate regions. Carbohydrates include monosaccharides, disaccharides, oligosaccharides, and polysaccharides. Rhamnose is the only monosaccharide that results in low CH4 emissions. However, rhamnose is a minor component in most plants. Among polysaccharides, pectic polysaccharides lead to greater CH4 production due to the conversion of methyl groups to methanol and finally to CH4. Thus, the degree of methyl esterification of pectic polysaccharides is an important structural characteristic to better understand CH4 emissions. Apart from pectic polysaccharides, the chemical structure of other polysaccharides per se does not seem to affect CH4 formation. However, rumen physiological parameters and fermentation types resulting from digestion in the rumen of polysaccharides differing in the rate and extent of degradation do affect CH4 emissions. For example, low rumen pH resulting from the rapid degradation of readily fermentable carbohydrates decreases and inhibits the activities of methanogens and further reduces CH4 emissions. When a large quantity of starch is supplemented or the rate of starch degradation is low, some starch may escape from the rumen and the escaped starch will not yield CH4. Similar bypass from rumen digestion applies to other polysaccharides and needs to be quantified to facilitate the interpretation of animal experiments in which CH4 emissions are measured. Rumen bypass carbohydrates may occur in ruminants fed fresh forage, especially when the passage rate is high, which could be a result of high feed intake or high water intake. The type of carbohydrates affects the concentration of dissolved hydrogen, which consequently alters fermentation pathways and finally results in differences in CH4 emissions. We recommend that the degree of methyl esterification of pectic polysaccharides is needed for pectin-rich forage. The fermentation type of carbohydrates and rumen bypass carbohydrates should be determined in the assessment of mitigation potential.

Substituting ryegrass-based pasture with graded levels of forage rape in the diet of lambs decreases methane emissions and increases propionate, succinate, and primary alcohols in the rumen.

Feeding 100% forage rape to sheep consistently lowers methane emissions per unit of intake (CH4/DMI) compared to those fed 100% ryegrass pasture. However, forage rape is usually supplemented with other feeds, which might impact the mitigation potential provided by forage rape. The objective of this study was to determine the effect of substituting ryegrass with graded levels of forage rape in the diet of lambs on methane emissions and rumen fermentation characteristics. Seventy wether lambs (n = 14/treatment) were fed a ryegrass-based pasture substituted with 0%, 25%, 50%, 75%, and 100% of forage rape (Brassica napus; FR0, FR25, FR50, FR75, and FR100, respectively) on a dry matter basis. Methane emissions and dry matter intake were measured for 48 h in respiration chambers and a rumen fluid sample was collected. CH4/DMI decreased (P < 0.01) with increasing forage rape inclusion in the diet so that sheep fed FR100 and FR75 emitted 34% and 11% less, respectively, than those fed FR0. CH4/DMI differences for lambs fed FR25 and FR50 were much smaller (<6%) relative to FR0. The pH of rumen fluid decreased (P < 0.01) at higher levels of forage rape inclusion in the diet (FR75 and FR100) compared to low levels of inclusion (FR0, F25, and F50). The proportion of ruminal acetate was least in FR100 (30%) followed by FR75 (10%), FR50 (8%), and FR25 (4%) compared with FR0 (P < 0.001). The proportion of propionate plus succinate was greater for FR100 (+40%), FR75 (+28%), and FR50 (+29%) compared with FR0, with FR25 intermediate (P < 0.001). The methanol concentration, and ethanol and propanol proportions in rumen fluid were greater for FR100 compared with any other treatment (P < 0.001). In conclusion, CH4/DMI decreased at high levels of forage rape inclusion in the diet and especially feeding FR100 was associated with a pronounced shift in rumen fermentation profile, with a significant presence of succinate, ethanol, propanol, methanol, valerate, and caproate.

The methane yield (g methane/kg dry matter intake) was 34% lower in sheep fed 100% forage rape and 11% lower in sheep fed 75% forage rape compared to sheep fed 100% ryegrass-based pasture. Sheep fed 25% and 50% forage rape as part of their diet had similar methane yields to sheep fed 100% ryegrass pasture. Sheep fed 100% forage rape had a ruminal fermentation profile with a smaller proportion of acetate and greater proportions of fermentation products like propionate, succinate, and valerate. Acetate formation is associated with hydrogen gas formation, which in turn is converted to methane in the rumen. Propionate, succinate, and valerate are alternatives to acetate plus hydrogen production and so fermentation shifts to them result in less methane formation.

Post-treatment bacterial endocarditis mimicking fungal organisms: a morphologic comparison and tips for avoiding this diagnostic pitfall.

BACKGROUND: Histopathologic differentiation of bacterial endocarditis from yeast-like fungal endocarditis is usually straightforward; however, an underappreciated phenomenon is the effect of antimicrobial therapy on bacterial size, shape and septa (cross-wall) formation resulting in bacterial forms that mimic yeast-like fungi. In this article we illustrate the alterations that occur in antibiotic-treated Staphylococcus aureus endocarditis and compare these changes to histopathologic findings in unaltered S. aureus and Histoplasma endocarditis, respectively. METHODS: Resected valves from three cases of endocarditis were compared based on the type ofinflammatory reaction, organism morphology and culture results. Case 1 was S. aureus endocarditis initially misclassified as Histoplasma due to its atypical morphologic and histopathologic features. The two cases included for comparison were an S. aureus endocarditis with more classic features and an Histoplasma capsulatum endocarditis. Hematoxylin and eosin (H&E), Gram, periodic acid Schiff (PAS), Gomori-Grocott methenamine silver stains (GMS), and culture results were compared in all cases. Molecular and immunohistochemistry tests were used for confirmation of first case. High power oil-immersion was used to visualize organisms' characteristics in all three cases. RESULTS: Case 1 and Case 3 (Histoplasma-infected valves) had fibrinous exudates with scattered macrophages. The microorganisms observed in the first case of methicillin-sensitive S. aureus (MSSA) were ∼ 2-3 µm by GMS stain and had prominent septations. Histoplasma yeast were round to oval, ∼ 3-4 µm in size and demonstrated budding. S. aureus without alterations were round, ∼ 1 µm in size, and lacked prominent septations. Necrotizing purulent inflammation was present in the unaltered case of MSSA. The MSSA case with alterations from antibiotic treatment did not stain well with the Gram stain and organisms were best visualized with the PAS and GMS stains. CONCLUSIONS: Antibiotic therapy for bacterial endocarditis can alter the inflammatory reaction to infection, bacterial size, septa formation, and staining characteristics. Knowledge of these therapy-related effects and use of high-power magnification helps to avoid misclassification as yeast-like fungi.

Abdominal normothermic regional perfusion in controlled donation after circulatory determination of death liver transplantation: Outcomes and risk factors for graft loss.

Postmortem normothermic regional perfusion (NRP) is a rising preservation strategy in controlled donation after circulatory determination of death (cDCD). Herein, we present results for cDCD liver transplants performed in Spain 2012-2019, with outcomes evaluated through December 31, 2020. Results were analyzed retrospectively and according to recovery technique (abdominal NRP [A-NRP] or standard rapid recovery [SRR]). During the study period, 545 cDCD liver transplants were performed with A-NRP and 258 with SRR. Median donor age was 59 years (interquartile range 49-67 years). Adjusted risk estimates were improved with A-NRP for overall biliary complications (OR 0.300, 95% CI 0.197-0.459, p < .001), ischemic type biliary lesions (OR 0.112, 95% CI 0.042-0.299, p < .001), graft loss (HR 0.371, 95% CI 0.267-0.516, p < .001), and patient death (HR 0.540, 95% CI 0.373-0.781, p = .001). Cold ischemia time (HR 1.004, 95% CI 1.001-1.007, p = .021) and re-transplantation indication (HR 9.552, 95% CI 3.519-25.930, p < .001) were significant independent predictors for graft loss among cDCD livers with A-NRP. While use of A-NRP helps overcome traditional limitations in cDCD liver transplantation, opportunity for improvement remains for cases with prolonged cold ischemia and/or technically complex recipients, indicating a potential role for complimentary ex situ perfusion preservation techniques.

Effect of dietary inclusion of winter brassica crops on milk production, feeding behavior, rumen fermentation, and plasma fatty acid profile in dairy cows.

This study determined feeding behavior, dry matter (DM) intake (DMI), rumen fermentation, and milk production responses of lactating dairy cows fed with kale (Brassica oleracea) or swede (Brassica napus ssp. napobrassica). Twelve multiparous lactating dairy cows (560 ± 22 kg of body weight, 30 ± 4 kg of milk/d, and 60 ± 11 d in milk at the beginning of the experiment; mean ± standard deviation) were randomly allocated to 3 dietary treatments in a replicated 3 × 3 Latin square design. The control diet comprised 10 kg of grass silage DM/d, 4 kg of ryegrass herbage DM/d, and 8.8 kg of concentrate DM/d. Then, 25% of herbage, silage, and concentrate (DM basis) was replaced with either kale or swede. Cows offered kale had decreased total DMI compared with cows fed the control and swede diets, whereas inclusion of swede increased eating time. Milk production, composition, and energy-corrected milk:DMI ratio were not affected. Cows fed with kale had a greater rumen acetate:propionate ratio, whereas swede inclusion increased the relative percentage of butyrate. Estimated microbial N was not affected by dietary treatments, but N excretion was reduced with inclusion of kale, improving N utilization. Cows fed kale tended to have increased nonesterified fatty acids and showed presence of Heinz-Ehrlich bodies, whereas hepatic enzymes such as aspartate aminotransferase, γ-glutamyl transferase, and glutamate dehydrogenase were not affected by dietary treatments. In plasma, compared with the control, swede and kale reduced total saturated fatty acids and increased total polyunsaturated fatty acids and total n-3 fatty acids. Overall, feeding cows with winter brassicas had no negative effect on production responses. However, mechanisms to maintain milk production were different. Inclusion of swede increased the time spent eating and maintained DMI with a greater relative rumen percentage of butyrate and propionate, whereas kale reduced DMI but increased triacylglycerides mobilization, which can negatively affect reproductive performance. Thus, the inclusion of swede may be more suitable for feeding early-lactating dairy cows during winter.

Factors Related to Weight Gain in Subjects with Sleeve Gastrectomy During Lockdown by the COVID-19 Pandemic.

PURPOSE: The COVID-19 pandemic could exacerbate the risk factors for weight gain in patients with previous bariatric surgery. The aim of this study was to evaluate factors related to weight gain during lockdown in patients with a sleeve gastrectomy. MATERIALS AND METHODS: A group of 48 obese subjects with previous bariatric surgery was enrolled. After a 7-week confinement, a telephone interview was conducted. In this phone call, self-reported body weight gain and different factors were recorded. In order to obtain the basal and pre-surgical data, biochemical and anthropometric parameters were recorded from electronic medical record. RESULTS: The mean age was 45.3±8.0 years (range: 23-61) and the mean body mass index (BMI) was 32.5±7.5 kg/m2 (range: 28.6-34.2). Gender distribution was 38 females (79.2%) and 10 males (20.8%). The increase in self-reported body weight was 3.8±2.1 kg during the 7 weeks of confinement. And the self-reported body weight gain was lower in subjects with regular exercise (4.6±0.9 vs 1.1±0.3 kg; p=0.02). The number of face-to-face visits to the nutrition office that did not attend was 0.61±0.81 (range: 0-4) per patient. In the multiple regression analysis with self-reported body weight gain as a dependent variable, the physical activity (minutes/week) remained as a protective factor with a beta coefficient of -0.09 (95% CI: -0.001 to 0.016; p=0.03) and number of face-to-face appointments in the nutrition consultation missed as a risk factor with a beta coefficient of 9.65 (95% CI: 1.17-18.12; p=0.03). CONCLUSIONS: The increase in self-reported body weight is associated with a decrease in physical activity and the loss of face-to-face visits to the Nutrition Unit.

Preparation, Assembly, and Transduction of Transgenic Elements Using Mosaic Analysis with Dual Recombinases (MADR).

This protocol focuses on the cloning and stable integration of sequences of interest by the use of a mosaic analysis with dual recombinases (MADR) plasmid that includes fusion proteins or independent proteins under the control of 2A peptide or IRES elements. Additionally, we describe how to generate a neural stem cell culture from Gt(ROSA)26Sortm4(ACTB-tdTomato, EGFP)Luo/J mice, and validate the MADR plasmids in vitro and in vivo by neonatal mouse brain electroporation. This protocol can be generalized to analyze any transgenic element using MADR technology. For complete details on the use and execution of this protocol, please refer to Kim et al. (2019).

A circadian rhythm-related MTNR1B genetic variant (rs10830963) modulates glucose metabolism and insulin resistance after body weight loss secondary to biliopancreatic diversion surgery.

INTRODUCTION: Objective: the rs10830963 SNP of the MTNR1B gene may be related with biochemical changes after weight loss induced by caloric restriction. We investigated the role of this SNP on biochemical parameters after biliopancreatic diversion (BPD) surgery in morbid obese subjects. Patients and methods: one hundred and fifty-four patients with morbid obesity, without diabetes mellitus type 2, were enrolled. Their biochemical and anthropometric parameters were recorded before the procedure and after one, two, and three years of follow-up. All subjects were genotyped (rs10830963) at baseline. Results: the decrease in fasting insulin levels seen after the first year (delta: -3.9 ± 1.2 mIU/L vs. -1.8 ± 1.1 mIU/L; p = 0.03), the second year (delta: -5.0 ± 0.3 mIU/L vs. -2.3 ± 0.2 mIU/L; p = 0.01) and the third year (delta: -5.1 ± 1.9 mIU/L vs. -2.8 ± 1.1 mIU/L; p = 0.02) was higher in non-G-allele carriers than in G-allele carriers. Additionally, the improvement of HOMA-IR levels at year one (delta: -0.7 ± 0.2 mIU/L vs. -0.2 ± 0.2 mIU/L; p = 0.03), year two (delta: -1.0 ± 0.3 mIU/L vs. -0.5 ± 0.2 mIU/L; p = 0.01) and year three (delta: -1.2 ± 0.3 mIU/L vs. -0.4 ± 0.2 mIU/L; p = 0.03) was also higher in non-G-allele carriers than in G-allele carriers. Finally, basal glucose levels after the first year (delta: -10.1 ± 2.4 mg/dL vs. -3.6 ± 1.8 mg/dL; p = 0.02), the second year (delta: -16.0 ± 2.3 mg/dL vs. -8.4 ± 2.2 mg/dL; p = 0.01) and the third year (delta: -17.4 ± 3.1 mg/dL vs. -8.8 ± 2.9 mg/dL; p = 0.03) were higher in non-G-allele carriers than in G-allele carriers, too. Improvements seen in comorbidities were similar in both genotype groups. Conclusion: our study showed an association of the rs10830963 MTNR1B polymorphism after massive weight loss with lower glucose response, insulin resistance, and fasting insulin levels in G-allele carriers.

INTRODUCCIÓN: Objetivo: la variante SNP rs10830963 del gen MTNR1B podría estar relacionada con cambios bioquímicos tras la pérdida de peso inducida por una restricción calórica. El objetivo de este trabajo es evaluar el papel de este SNP en los parámetros bioquímicos después de la cirugía de derivación biliopancreática (DBP). Pacientes y métodos: se reclutaron un total de 154 pacientes con obesidad mórbida sin diabetes mellitus de tipo 2. La valoración bioquímica y antropométrica se realizó antes de la intervención y tras 1, 2 y 3 años de seguimiento. Todos los sujetos fueron genotipados (rs10830963) en el momento basal. Resultados: la disminución de los niveles de insulina en ayunas después del primer año (delta: -3,9 ± 1,2 mUI/L vs. -1,8 ± 1,1 mUI/L; p = 0,03), el segundo año (delta: -5,0 ± 0,3 mUI/L vs. -2,3 ± 0,2 mUI/L; p = 0,01) y el tercer año (delta: -5,1 ± 1,9 mUI/L vs. -2,8 ± 1,1 mUI/L; p = 0,02) fueron mayores en los no portadores del alelo G que en los portadores. Además, la mejora de los niveles de HOMA-IR en el primer año (delta: -0,7 ± 0,2 mUI/L ± -0,2 ± 0,2 mUI/L; p = 0,03), segundo año (delta: -1,0 ± 0,3 mUI/L vs. -0,5 ± 0,2 mUI/L; p = 0,01) y en el tercer año (delta: -1,2 ± 0,3 mUI/L vs. -0,4 ± 0,2 mUI/L; p = 0,03) también fueron mayores en los no portadores del alelo G. Finalmente, los niveles basales de glucosa después del primer año (delta: -10,1 ± 2,4 mg/dL vs. -3,6 ± 1,8 mg/dL; p = 0,02), el segundo año (delta: -16,0 ± 2,3 mg/dL vs. ­ 8,4 ± 2,2 mg/dL; p = 0,01) y el tercer año (delta: -17,4 ± 3,1 mg/dL vs. -8,8 ± 2,9 mg/dL; p = 0.03) fueron mayores en los no portadores del alelo G. Las comorbilidades mejoraron en ambos genotipos de manera similar. Conclusión: nuestro estudio mostró una asociación del polimorfismo rs10830963 MTNR1B tras una pérdida de peso posquirúrgica con una menor respuesta de los niveles de glucosa, resistencia a la insulina e insulina en ayunas en portadores del alelo G.