bluuebunny/crossref_metadata_2025 huggingface dataset:
White tepary bean shows higher in vitro iron bioavailability than brown tepary or common bean
Authors: Bries Amanda E. , Reddy Manju B. , Winham Donna M.
Published: November 2023

Iron deficiency anemia is the most prevalent global micronutrient disorder, especially in populations who consume mainly plant‐based diets. Although beans contain a greater amount of iron in comparison to wheat, corn or other staple crops, the bioavailability is often low due to inhibitory factors like polyphenols and phytic acid. Tepary beans (Phaseolus acutifolius) thrive better in arid climates than other beans, making them a suitable crop for expansion into marginal ecological zones with climate change. Iron bioavailability is unknown for many tepary varieties, but proximate analysis shows higher iron content than in most common bean (Phaseolus vulgaris) varieties. The study purpose was to determine in vitro iron bioavailability, proximate analysis, polyphenol, and phytic acid content of tepary and common bean varieties. Increased iron bioavailability in a bean with arid climate resiliency can offer a sustainable solution towards reducing iron deficiency anemia in developing countries. Whole seeds of 5 tepary white, 3 tepary brown, 4 black, and 2 pinto bean varieties were analyzed. Polyphenol (PP) and phytic acid (PA) extractions were assessed colorimetrically. Iron concentrations were determined using ICP‐mass spectrometry following a microwave digestion. Iron solubility analysis was completed through in vitro digestion of the cooked beans. Supernatants from the in vitro digestion were used in determining the iron bioavailability through a CaCo‐2 cell culture model. For statistical analysis, results were pooled by tepary white, tepary brown, black bean, and pinto bean. Both tepary white and tepary brown groups showed significant differences in PP content in comparison to the black and pinto (p≤0.05), whereas PA, amongst all groups, showed no significant differences (p>0.05). Iron content ranged from 29.8 μg/g to 78.47 μg/g, with pinto beans having significantly lower iron concentration (mean = 33.64 μg/g). Significant differences in percent solubility were found between the pinto bean and tepary varieties (p≤0.05), but not the black. Iron bioavailability of the tepary white showed a negative correlation with PP content (high iron, low PP). The tepary white showed significantly higher iron bioavailability (p≤0.02) in comparison to both pinto and black beans. Our data suggests that the low PP and PA contributes to higher iron bioavailability in the white varieties of Phaseolus acutifolius. Further studies to replicate these findings are needed, followed by clinical testing of tepary white iron bioavailability in humans.
Effects of Selenium in Comparison to Exendin‐4 on the Expression of GLP‐1R, IRS‐1 and Preproinsulin in the Pancreas of Diabetic Rats
Authors: Rizk Sandra , Barakat Ghinwa , Moustafa Mohamed , Khalifeh Ibrahim , Hodroj Mohammad H , Bikhazi Anwar
Published: November 2023

Antioxidant supplements are considered as being favorable for the treatment of diabetes mellitus. There is a debate regarding the effect of selenium on the incidence of diabetes mellitus: even though some clinical trials studies suggested that high selenium supplementation resulted in an increased risk of incidence of diabetes mellitus, other studies showed that the level of serum selenium in diabetic patients was lower than its level in nondiabetic subjects, and high selenium status may exert a protective action against the incidence of diabetes mellitus. Exendin‐4 is a GLP‐1 agonist with antidiabetic properties. It binds and activates GLP‐1R with greater potency than GLP‐1 and is used in the treatment of diabetes mellitus. The mechanisms by which selenium and exendin‐4 exert their antidiabetic actions are still unclear. In this study, we investigated the effects of selenium or exendin‐4 administration on the expression of glucagon‐like peptide‐1 receptor (GLP‐1R), insulin receptor substrate‐1 (IRS‐1), and preproinsulin in the pancreas of diabetic rats. Diabetes was induced by streptozotocin administration. Diabetic rats were injected intraperitonially with 0.03 μg exendin‐4/kg body weight/daily or treated with 5 ppm selenium in drinking water for a period of 4 weeks. Results obtained indicate that GLP‐1R and IRS‐1 levels decreased while the level of preproinsulin mRNA increased in the pancreas of diabetic untreated rats, as compared to control rats. Treatment of diabetic rats with exendin‐4 increased protein and mRNA levels of GLP‐1R, and IRS‐1, as well as the mRNA level of preproinsulin in the pancreas, as compared to their levels in diabetic untreated rats. Selenium treatment of diabetic rats increased the pancreatic mRNA levels of GLP‐1R, IRS‐1, and preproinsulin. The numbers of pancreatic islets and GLP‐1R molecules in the pancreas of diabetic rats was also increased upon treatment with selenium or Exendin‐4. Therefore, exendin‐4 and selenium may exert their antidiabetic effects by increasing GLP‐1R, IRS‐1, and preproinsulin expression in the pancreas and by increasing the number of pancreatic islets.
The Breath Carbon Isotope Ratio is a Promising Biomarker of Added Sugar Intake
Authors: O'Brien Diane M , Black Jynene A , Niles Kristine , Schoeller Dale
Published: November 2023

High consumption of added sugars in the US is a significant public health concern. Added sugar intake is widely misreported, hampering efforts to understand its association with health. The carbon stable isotope ratio has been proposed as a biomarker of added sugar intake, because it is elevated in corn and sugar cane, the source of most added sugars consumed in the US. The objective of this study was to investigate the carbon isotope ratio of breath as a biomarker of short term (same day) added sugar consumption. Breath can be collected quickly and non‐invasively, and breath carbon isotope ratios can be rapidly measured using cavity ring‐down spectroscopy (CRDS). If shown to be a valid measure of short term added sugar intake, the breath carbon isotope ratio could be feasibly collected and measured multiple times, then averaged to estimate usual intake. Our objective was to test whether breath carbon isotope ratios could distinguish low (0 g/day), medium (75 g/day) and high (150 g/day) added sugar intakes, using a randomized cross‐over study of 12 adults (6 female, 6 male). Diets were 50% carbohydrate, 30% fat, and 20% protein, and included breakfast and lunch only (delivering 60% of daily energy and added sugar target values). On each study day, participants provided breath samples by blowing into a gas sampling bag at 8 am (fasted), 10 am, 12 pm, 2 pm and 4 pm, and were served breakfast and lunch following the 8 am and 12 pm sampling times, respectively. There were 1–2 weeks between each treatment day. The effect of added sugar level on breath carbon isotope ratios was analyzed for each sampling time point using Analysis of Covariance, including sex, age, and fasting breath isotope ratios as covariates. Breath carbon isotope ratios increased significantly with added sugar intake, and differed significantly between all three treatments (low, medium and high added sugar, all P < 0.0001) at all four post‐feeding time points (10 am, 12 pm, 2 pm and 4 pm). Fasting breath carbon isotope ratios were a significant covariate, especially for the 10 am and 12 pm measurements. This preliminary work shows that breath carbon isotope ratios are promising measures of short term sugar intake that are non‐invasive and easily‐collected. If collected over multiple days and averaged, these measures could potentially provide objective biomarkers of usual added sugar intake for epidemiological research.

Support or Funding Information

P30 GM 103325‐03
Energy Deficiency, but not Aerobic Exercise, is Associated with Increased Stable Nitrogen Isotope Ratio of Urinary Urea
Authors: Koehler Karsten , Flenker Ulrich , Schaenzer Wilhelm , Huelsemann Frank
Published: November 2023

Energy deficiency (ED) is typically associated with the loss of lean mass, which occurs secondary to increased body protein breakdown. Exercise, which stimulates both protein synthesis and breakdown, has the capacity to attenuate the loss of lean mass during ED, but the contribution of protein sources, i.e. body protein vs. dietary protein, to protein turnover during ED and/or exercise remains only poorly understood. Analysis of the stable nitrogen isotope ratio (δ15N) of urea, the primary end‐product of protein metabolism, could provide useful information about the source of protein breakdown and, subsequently, for dietary protein requirements to preserve lean mass during ED.

PURPOSE

To determine the effects of ED with and without exercise on the stable nitrogen ratio of urinary urea (δ15Nurea).

METHODS

Healthy, trained, young men (n=6, 25.2 ± 2.4 y, 79.7 ± 7.7 kg; 9.6 ± 1.5% body fat) were included in this 4‐way crossover study. Participants underwent two 4‐day ED conditions during which energy availability (EA) was reduced to 15 kcal/kg fat free mass (FFM) per day, once while conducting exercise to expend 15 kcal/kg FFM (ED+EX) and once while not exercising (ED‐EX). Participants also underwent two 4‐day control conditions in energy balance (EA of 40 kcal/kg FFM), once with exercise (C+EX) and once without exercise (C‐EX). δ15Nurea was assessed from urine samples using elemental analysis‐isotope ratio mass spectrometry (EA‐IRMS, Eurovektor EA 3000) and was adjusted for dietary nitrogen isotope ratio (δ15Ndiet), which was computed as previously described [1].

RESULTS

Weight loss was similar in both ED conditions (ED‐EX: −2.4±0.7 kg, p<0.001; ED+EX: −1.8±1.0 kg, p=0.004). δ15Nurea adjusted for δ15Ndiet (δ15Nurea ‐δ15Ndiet) was significantly elevated in ED‐EX when compared with C‐EX (0.3±0.4‰ vs. −1.1±0.4‰, p=0.01) and in ED+EX when compared with C+EX (−0.6±1.0‰ vs. −1.1±0.4‰, p=0.01); whereas, there was no significant difference in δ15Nurea ‐δ15Ndiet between ED‐EX and ED+EX and between C‐EX and C+EX. Multiple linear regression identified EA (p=0.015), dietary protein intake (p=0.001), and the interaction between EA and protein intake (p=0.03) as significant predictors of δ15Nurea ‐δ15Ndiet.

CONCLUSION

The increase in δ15Nurea during both ED conditions confirms that endogenous protein breakdown is increased in the energy‐deficient state. However, our results do not indicate that exercise per se is associated with changes in endogenous protein breakdown. Future studies with controlled dietary protein intake are needed to verify that changes in δ15Nurea are reflective of increased body protein breakdown during ED.

Support or Funding Information

The study was supported by a University seed grant.
Determinants of Developmental Milestones in Infants and Young Children in Bihar, India
Authors: Larson Leila , Mehta Rukshan , Kekre Priya , Verma Pankaj , Young Melissa , Girard Amy Webb , Ramakrishnan Usha , Chaudhuri Indrajit , Srikantiah Sridhar , Martorell Reynaldo
Published: November 2023

Approximately 200 million children globally fail to fulfill their developmental potential. Children in Bihar, India may be at particularly high risk as the extensive malnutrition in the region could have important implications for brain development. The objectives of this study are to use baseline data from an ongoing cluster randomized effectiveness trial of home fortification with micronutrient powders in Bihar to 1) validate a modified version of the Developmental Milestones Checklist II (DMC‐II) and 2) determine nutritional and socioeconomic risk factors for mental and motor development of infants and young children.

Cross‐sectional data from 4360 children 6–18 months of age living in West Champaran district of Bihar, India, were used to determine the predictive validity of the modified DMC‐II by comparing scores by age, nutritional status, and maternal education. Multivariate regression was used to examine determinants of child development score.

The mean score on the modified DMC‐II was 40.9 (95%CI 40.6, 41.3) out of 82.0. The overall score and scores for each subscale (motor, language, personal‐social, and cognitive) were significantly associated with child age and maternal education. After adjustment for age, overall and all subscale scores were each significantly associated with height‐for‐age, weight‐for‐age, and weight‐for‐height z‐scores. A multivariate linear model for the child's overall modified DMC‐II scores (R2=0.60) revealed significant associations (P<0.05) with age (coefficient (SE): 2.49 (0.05)), gender (−0.84 (0.28)), height‐for‐age z‐score (0.57 (0.15)) weight‐for‐age z‐score (1.10 (0.17)), dietary diversity score (1.00 (0.14)), timely initiation of complementary foods (1.00 (0.29)), no use of prelacteal feeds (0.67 (0.29)), Family Care Indicators or home stimulation (0.56 (0.04)), and maternal education (0.97 (0.30)). Recent morbidity (fever, cough, or diarrhea) was not significantly associated with DMC‐II scores in the multivariate model.

Predictive validity of the DMC‐II was established in this population. Comparison of standardised regression coefficients identified weight‐for‐age z‐score, Family Care Indicators, and dietary diversity as the strongest determinants of the child's overall score on the developmental milestones; length‐for‐age z‐score, weight‐for‐age z‐score, and weight‐for‐length z‐score were the strongest predictors of score on the motor development subscale. Strategies to improve child development should simultaneously address these issues.

Support or Funding Information

Funding support provided by a BMGF grant through a subcontract with CARE‐India
Maternal Vitamin D Status and Adverse Pregnancy and Neonatal Outcomes in India
Authors: Finkelstein Julia L. , Duggan Christopher , Mehta Saurabh , Thomas Tinku , Srinivasan Krishnamachari , Kurpad Anura V.
Published: November 2023

Objective

To examine the prevalence of vitamin D deficiency and its association with risk of adverse pregnancy and neonatal outcomes in Southern India.

Methods

A total of 366 women participating in a vitamin B12 supplementation trial in India were monitored to assess plasma 25‐hydroxyvitamin D (25OHD) concentrations in each trimester during pregnancy. All women received daily iron and folic acid supplementation during pregnancy. Binomial and linear regression models were used to evaluate the associations of maternal vitamin D status and pregnancy and neonatal outcomes.

Results

Vitamin D insufficiency was common in pregnancy: 85% of mothers were vitamin D deficient (25OHD <20.0 ng/mL), and 40% had vitamin D concentrations less than 10.0 ng/mL at their first prenatal visit. Maternal vitamin D concentrations increased significantly during pregnancy (p<0.05). However, 75% of mothers were vitamin D deficient (25OHD <20.0 ng/mL) and 31% had vitamin D concentrations less than 10.0 ng/mL in the third trimester. Maternal vitamin D status was not significantly associated with risk of adverse pregnancy outcomes, including infant birth weight, low birth weight (<2,500 g), intrauterine growth restriction, gestational age at delivery, or preterm delivery (<37 weeks). However, maternal vitamin D concentrations at baseline were associated with increased neonatal head circumference (p=0.02) in multivariate analyses, after adjusting for gestational age at sample collection, vitamin B12 intervention, maternal body mass index (BMI), socioeconomic status, and C‐reactive protein concentrations. Maternal vitamin D deficiency at enrollment was associated with significantly lower neonatal mid‐upper arm circumference (p=0.02) and biceps skinfold thickness (p=0.04) in multivariate analyses, after adjusting for gestational age at sample collection, vitamin B12 intervention, maternal BMI, socioeconomic status, and C‐reactive protein concentrations.

Conclusions

Vitamin D deficiency is common in pregnant Indian women, and is associated with lower neonatal arm anthropometric and fat measures.

Support or Funding Information

National Institutes of Health, Indian Council of Medical Research, Harvard University
Alternation of Nephron Morphology Associated with Sustained Delivery of Danazol Using Adult Male Rodents as a Model
Authors: Benghuzzi Hamed A , Tucci Michelle A
Published: November 2023

Hormonal replacement therapy (HRT) has shown to be efficacious in treatment and preventing of heart disease, osteoporosis and reducing mortality in hypogonadal males. Several attempts to utilize the native testosterone and its analog such as danazol have shown different physiological responses. In addition, the route of administration and its mode of action is lacking in the literature. The specific objective of this study was to investigate the role of sustained delivery of danazol on the functional and structural capacity of the kidney using adult male rats as a model. A total of 24 adult male rats were subdivided into four equal groups. Groups I and II were castrated by following standard laboratory surgical procedures. Each rat in groups II and III (intact) were implanted with tricalcium phosphate lysine (TCPL) drug delivery system loaded with 40 mg of danazol. Rats in group IV were unimplanted and untreated to be served as a control group. At the end of 90 days post treatment the animals were sacrificed by using overdose of isoflurane and assured by cervical dislocation. Vital and reproductive organs were retrieved, weighed and subjected to H&E staining procedure. The results of this investigation suggest: (i) TCPL delivery system released danazol at a sustained level for 60 days without any untoward response, (ii) the wet weights of kidneys (normalized to body weight) were increased (p<0.05) in intact rats treated with danazol compared to control, (iii) sustained delivery of danazol resulted in a maintenance of kidney weights compared to the control level, however, the lack of danazol treatment resulted in a remarkable regression in the kidney weights of castrated rats, (iv) histopathological evolutions of renal tubules revealed tubular epithelial damage in intact rats received danazol treatment compared to the control and other groups. In conclusion, this study suggests that exogenous danazol therapy could lead to profound tubular damage and consequently major renal insufficiency.
Maternal Obesity, 25(OH)D Concentration and Bone Mineral Density in Breastfeeding Dyads
Authors: Sen Sarbattama , Penfield‐Cyr Annie , Lu Zhigang , Ebeling Myla , Hollis Bruce , Howard Cindy , Wagner Carol
Published: November 2023

Background

Previous studies have shown inverse relationships between Body Mass Index (BMI) and serum levels of vitamin D. Among other sequelae, vitamin D deficiency is related to bone health risks including rickets in children and osteomalacia in adults. The impact of maternal BMI on vitamin D levels and bone mineral density (BMD) in breastfeeding mother‐infant dyads has not been studied.

Objective

To examine the association between maternal BMI and vitamin 25(OH)D concentration and BMD in exclusively breastfeeding mother‐infant pairs.

Methods

We conducted a secondary analysis of a cohort of 278 breastfeeding mother‐infant dyads who were recruited in the first month post‐partum. Using ANOVA and linear regression, we examined associations between maternal BMI continuously and categorically (Lean (L), BMI 18.5–25kg/m2; overweight (Ow), BMI 25–30kg/m2; and Obese (Ob), BMI≥30kg/m2) with four outcomes: Maternal and infant 25(OH)D concentration, maternal and infant BMD using DXA.

Results

Mean maternal BMI was 28.5 kg/m2. Maternal BMI negatively associated with maternal serum concentration of 25(OH)D (β=−0.29 nmol/L decreased in 25(OH)D concentration for each increase in BMI point, p<0.0001), with Ob mothers having significantly lower mean levels than L or Ow (mean±SD nmol/L: L: 38.4±14.6, Ow: 34.2±14.3, Ob: 29.4±9.9; p<0.05 for Ob vs. L and Ob vs. Ow). Despite lower serum 25(OH)D concentrations, Ob women had higher BMD than either L or Ow women (L: 0.97±0.14g/cm3, Ow 1.01±0.11g/cm3, Ob 1.05±0.11g/cm3; p<0.05 for Ob vs. L and Ob vs. Ow). There was no association between maternal BMI and infant serum 25(OH)D concentration or infant BMD (p=0.5 and 0.98 respectively).

Conclusions

Maternal obesity was associated with lower serum 25(OH)D concentration in breastfeeding women, but not infant serum 25(OH)D concentration or infant BMD. There seems to be a protective effect of obesity on maternal bone mineralization at this early time point in lactation. The effect of prolonged 25(OH)D deficit throughout lactation on maternal and infant bone mineralization is currently being analyzed.

Support or Funding Information

Funded in part by National Institutes of Health (NIH) 5R01HD043921, NIH RR01070, Medical University of South Carolina Department of Pediatrics, and by the South Carolina Clinical & Translational Research (SCTR) Institute, with an academic home at the Medical University of South Carolina, NIH/National Center for Advancing Translational Sciences grant UL1 TR000062, and a K23 grant from the NIH (ref. no. K23HD074648).
TMPRSS4: a novel serine protease involved in IPF development?
Authors: Valero Ana Mayela , Cisneros José , Ramírez Remedios , Gaxiola Miguel , Becerril Carina , Pardo Annie , Selman Moisés
Published: November 2023

Background

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive and lethal lung disease characterized by the expansion of the fibroblast population and excessive extracellular matrix accumulation with the subsequent destruction of the lung architecture. Its pathogenesis is incompletely understood but evidence suggests an epithelial pathway in which IPF may result from epithelial microinjuries and activation and abnormal wound healing. TMPRSS4 is a novel type II transmembrane serine protease found at the cell surface. Although their biological functions are unknown it has been suggested that TMPRSS4 promotes migration and facilitates the epithelial to mesenchymal transition (EMT), two critical processes in the pathogenesis of IPF. Thus, we hypothesized that overexpression of TMPRSS4 in the lung could promote the initiation and/or progression of IPF.

Aim

To determine the role of TMPRSS4 in IPF.

Methods

We first examined the lung expression and localization of TMPRSS4 in IPF and normal lungs by qPCR and immunohistochemistry. Additionally, we measured the expression of TMPRSS4 in human lung fibroblasts and one alveolar and one bronchial epithelial cell lines. Then we explored the role of this protease in an experimental model of lung fibrosis. TMPRSS4 haplodeficient and wild type mice received an intratracheal instillation of bleomycin and were studied at 28 days post‐instillation.

Results

We found that this protease is not expressed in normal human lungs while variable amounts of the mRNA were found in IPF lungs (2−DCT 1.1×10−2 ± 9.4×10−5). TMPRSS4 was primarily expressed by epithelial and mast cells. The protease was not found in lung fibroblasts but different levels of mRNA were revealed in alveolar (2−DCT 7.5×10−7 ± 2×10−7) and bronchial (2−DCT 4.2×10−5 ± 3.6×10−6) epithelial cells. The lung fibrotic response, measured by hydroxyproline ( 178.9 ± 12.4 versus 110.7 ± 13.8 mg/lung; p<0.05) and morphology (40% versus 15% extent of the lesions) was significantly attenuated in the haplodeficient mice.

Conclusions

Our findings indicate that TMPRSS4 may have a profibrotic role in IPF and other fibrotic lung disorders.

Support or Funding Information

Posgrado en Ciencias Biológicas, UNAM; CONACYT SALUD‐2014‐01‐234088
Digital Determination of Chemotherapeutic Drug‐Induced Damage Sites in Single DNA Molecules using Nanofluidic Channels
Authors: Kaufman David Gordon , Chastain Paul David , Soper Steven Allan
Published: November 2023

We have shown that DNA can be extended on glass slides permitting the observation of fluorescently labeled DNA fibers and facilitating the quantitation of their length. When the stretched DNA fibers are also immunostained for the presence of apurinic/apyrimidinic (AP) sites, it was possible to quantitate the number of AP sites per unit length of DNA. Measurements of AP site frequency that are determined in this manner are comparable in accuracy to the “gold standard” method of AP site quantitation, but have a lower variance and the ability to analyze even single DNA molecules. Using this method, it has been possible to detect the increased frequency of AP site formation in DNA of cells exposed to hydrogen peroxide in cell culture. Furthermore, it was possible to show that DNA located at replication sites were more intensely damaged than the remainder of the DNA. Similar increases in the quantity of DNA damage sites have also been found in the DNA of cells treated with the cancer chemotherapeutic drug doxycycline. While this technique appears to be a powerful method to detect DNA damage in cells exposed to damaging agents in culture and in vivo, it has the liability of being extremely labor intensive and requires an extended time (weeks to days) for completion. To remedy these shortcomings and to allow clinical translation of this technology, we have developed an alternate approach, which involves the extraction of DNA from circulating tumor cells isolated from whole blood using a unique microfluidic device, immunostaining the DNA and the associated DNA AP sites in micro‐scale reaction chambers and the alignment and deposition of individual DNA fibers in nanochannels (<100 nm in width and depth). This enables single DNA molecules to be stretched reproducibly in nanochannels to near their full contour length which facilitates quantification of labeled AP sites. The DNA fibers positioned within the nanochannels are fluorescently imaged and the images subjected to a unique image analysis technique to provide highly quantitative information on AP site frequency. When using this image analysis approach, it is possible to obtain statistically significant results from even single cells. The combination of these microfluidic and nanofluidic devices into a single analysis platform for quantitating rates of DNA damage reduces the time required for this measurement to an hour or less, reduces technical time to that needed to load samples and reagents, and should permit these analyses to be made in the clinical setting by clinical laboratory personnel.

Support or Funding Information

Supported by NIH Grants R21‐125337 and P41‐EB020594.
Role of Sumoylation in Aldehyde Dehydrogenase 2 (ALDH) function during Alcoholic Liver Disease
Authors: Cossu Carla , Ramani Komal , Spissu Ylenia , Floris Andrea , Tomasi Maria Lauda
Published: November 2023

Background

Chronic alcohol abuse causes liver disease that progress from steatosis through stages of steatohepatitis, fibrosis, cirrhosis and eventually hepatic failure. Acetaldehyde, a major toxic ethanol metabolite, is mutagenic and carcinogenic, playing an important role in the pathogenesis of alcoholic liver disease (ALD) inducing production of reactive oxygen species (ROS) and apoptosis. ALDH2 is the major enzyme that metabolizes acetaldehyde produced from alcohol metabolism. Sumoylation is a posttranslational modification able to modify the activity, localization, and expression of the target protein by a covalent bind where small ubiquitin modifier (SUMO) is a tag. Increased ROS production enhances the global protein SUMO conjugation profile. We previously demonstrated that the level of Ubiquitin‐conjugation enzyme 9 (Ubc9), a sole E2‐conjugating enzyme essential for sumoylation, is induced in the livers of intragastric ethanol‐infusion (EI) treated mice. Our aim was to examine whether the dysregulated sumoylation could regulate the ethanol‐induced ALDH2 function in ALD and elucidate the molecular mechanism(s).

Methods

Studies were done using in vivo binge ethanol‐fed mice and primary mouse hepatocytes. Sumoylation protein profiling and expression were measured by Mass Spectrometry and Western blot analyses, respectively. ALDH2 activity was analyzed using commercial kit.

Results

We found that ALDH2 is sumoylated in binge mice livers analyzing the peptide‐sequencing data from sample purified by SUMO immunoaffinity columns. In addition, we found that both Ubc9 and ALDH2 mRNA and protein levels are increased in the livers of binge mice and ethanol‐treated hepatocytes. Interestingly, Ubc9 silencing prevented ethanol‐mediated induction in ALDH2 protein level. ALDH2 activity assay showed that Ubc9 is required for ALDH2 to be enzymatically active in primary mouse hepatocytes. Finally, we confirmed the peptide‐sequencing data of sumoylated ALDH2 using in vivo binge mice livers finding increased level of SUMO1/2/3‐ALDH2 complex formation following ethanol treatment.

Conclusions

We report for the first time that ethanol‐mediated sumoylation increased ALDH2 protein level and enzymatic activity in livers of binge mice and primary hepatocytes. This novel finding thereby opens a new area of investigation examining the importance of ALDH2 sumoylation in alcoholic liver disease.

Support or Funding Information

NIAAA Grant: 5 K01 AA022372‐04
Role of TLR7 in development of alcoholic fatty liver disease: a new target for prevention of alcoholic fatty liver disease
Authors: Matsumoto Hiroshi , Yang Chipin , Sugimoto Kana
Published: November 2023

TLRs play an important role in development of alcoholic fatty liver disease. Especially, TLR4 contributes to promote alcoholic fatty liver disease via priming via LPS. Recently, it has been reported that the treatment of a TLR7 ligands induces fatty liver disease. It has been recognized circulating microRNAs in experimental alcoholic fatty liver disease. TLR7 recognizes RNA to lead activation of innate immune system. Then we hypothesized that TLR7 contributes to develop alcoholic fatty liver disease. Lieber‐DiCarli alcohol‐diet pair feeding for 4 weeks performed in TLR7 knockout and wild mice. CpG, a ligand of TLR7, was treated in both mice. TLR7 deletion improved fatty liver injuries in histopathological findings, while CpG treatment promoted fatty liver injuries in TLR knockout mice. Therefore, these findings show that TLR7 contributes to develop fatty liver disease, indicating that regulation of TLR7 activation may prevent development of alcoholic fatty liver disease.

Support or Funding Information

A part of Grands‐in‐Aid for Scientific Reserch (B) (Grant #24390175) by Japan Society for the Promotion of Science (JSPS)
Caveolin‐1 regulation of DISC1 as a potential therapeutic target for schizophrenia
Authors: Kassan Adam , Egawa Junji , Zhang Zheng , Lajevardi Yasaman , Kim Kaitlyn , Nguyen Quynh My , Posadas Edmund S. , Sawada Atsushi , Jeste Dilip V. , Roth David M. , Patel Piyush M. , Patel Hemal H. , Head Brian P.
Published: November 2023

Background

Schizophrenia (SZ) is a debilitating neuropsychiatric disorder manifested in early adulthood. Disrupted‐In‐Schizophrenia‐1 (DISC1) is a promising candidate gene for SZ, but the molecular mechanisms underlying its role in the pathogenesis of the disease is still poorly understood. Caveolin‐1 (Cav‐1) is a cholesterol‐binding and plasma membrane scaffolding protein implicated in neuronal signal transduction and plasticity. Here we examined the role of CAV1 in mediating DISC1 expression in neurons in vitro and the hippocampus in vivo.

Methods and results

Overexpressing Cav‐1 in primary neonatal rodent neurons using a neuron specific synapsin promoter (SynCav1) isolated from hippocampi resulted in increased DISC1 and synaptic plasticity markers (PSD95, synaptobrevin, synaptophysin, and syntaxin‐1A). Similarly, SynCav1‐transfected differentiated human neurons derived from induced pluripotent stem cells (iPSCs) exhibited increased expression of DISC1 and markers of synaptic plasticity. Furthermore, these markers and DISC1 were significantly reduced in the hippocampi from Cav‐1 knockout (KO) mice. Finally, SynCav1 delivery to the hippocampus of Cav‐1 KO mice and neurons lacking Cav‐1 restored DISC1 protein expression as well as markers of synaptic plasticity.

Conclusion

These findings show an important role for neuronal Cav‐1 as a mediator of DISC1; therefore, SynCav1 might be a potential therapeutic target for SZ.

Support or Funding Information

5 T32 MH 19934‐20
LIPIDIC AGENT AS VASCULAR CONSTRAST‐MEDIUM FOR MRI (3T) IN FETAL AND EMBRYONIC DEVELOPMENT: FEASABILITY‐STUDY ON EX‐VIVO RABBIT FETUS AT 3T
Authors: TONNELET Romain , PEREZ Manuela , GONDIM TEIXEIRA Pedro Augusto , SEWONU Anou , BEAUMONT Marine , HOSSU Gabriela , LABROUSSE Marc , BRAUN Marc
Published: November 2023

Introduction

Understanding some dynamic processes such as the development of the cardiovascular system requires a global and precise anatomic vision of this dynamic and complex process

A variety of imaging tools and reconstructions are available for the evaluation of embryonic development. Micro‐MRI was used as the “standard” technique in large study series1, 2 competing with micro‐CT and techniques requiring the preparation of the sample in serial sections (EFIC method3 / 3D reconstruction from serial histological sections : EmbryoForm Project4).

Material & Method

Embryonic imaging is also restricted by the use of vascular MRI contrast agents such as gadolinium coupled to albumin, which are particularly expensive.

Lipidic agents have high viscosity at ambient temperatures and a T1 shortening effect for vascular rendering. Due to the non‐diffusion phenomenon related to its inherent hydrophobic properties, lipidic agents are restricted to the intra‐vascular compartment. These characteristics make lipidic agents a potentially available and cost effective option for embryonic vascular imaging.

In this study we sought to evaluate the use of a lipidic contrast agent (Type‐B oil / Fisher Scientific) using dedicated coils with a 3Tclinical MR scanner. A qualitative analysis of the injected vascular anatomical structures and a quantitative analysis by the contrast‐to‐noise ratio (CNR) and signal‐to‐noise ratio (SNR) was performed on two rabbit fetuses.

Results

Large and medium vessels structures including thoraco‐obdominal great vessels, intra‐hepatic vasculature and vascular brain structures were identified by two trained operators.

CNR and SNR measures were in agreement with those of the literature.

Such a technique have the potential to increase the availability of embryonic vascular imaging and facilitate research in the field of vasculogenesis.
The Interplay Between Taste and Chemesthesis
Authors: Huang Anthony Y , Wu Sandy Y
Published: November 2023

Chemosensory afferent fibers, which contribute to chemesthesis, the general chemical sensitivity of the mucus membranes in the oronasal cavities and being perceived as pungency, irritation or heat, innervating taste buds store transmitters such as calcitonin gene‐related peptide (CGRP) but little is known about their function. The complex interplay between taste cells and chemosensory neurons, via transmitters, may play an important role during the processing of the complex stimuli involving both spicy and tasty components. In this study, we characterized CGRP receptors present on taste cells. These receptors appear to underlie the postulated efferent transmission. Specifically, we loaded mouse vallate taste cells with Fura 2, conducted functional imaging, and used cellular biosensors to monitor taste‐evoked transmitter (ATP) release. The findings are that a significant fraction of Presynaptic (Type III) taste bud cells (53%) respond to CGRP, but that only few Receptor (Type II) taste cells (4%) are activated by this transmitter. We demonstrated that this effect of CGRP was dependent on phospholipase C activation. Consistent with these results, applying CGRP elicited taste buds to secrete serotonin (5‐HT), a Presynaptic (Type III) taste cell transmitter. Furthermore, CGRP‐evoked 5‐HT release inhibited taste buds and reduced their ability to secrete transmitter (ATP) during gustatory stimulation. We conclude that the net effect of CGRP as an efferent transmitter in taste buds is inhibitory—i.e., CGRP acts via serotonergic mechanisms to shape peripheral taste signals during processing gustatory information in taste buds.

Support or Funding Information

Supported by SIUSOM Research Seed Grant (AYH).
Middle and Inner Ear 3D Reconstructions using High Resolution X‐ray Micro‐Tomography
Authors: LABROUSSE Marc , DUBERNARD Xavier , SETTEMBRE Nicla , TONNELET Romain , AVISSE Claude , DEVY Jérôme , RENARD Yohann
Published: November 2023

Introduction

The temporal bone is a complex 3D structure that is difficult to teach to ENT residents. Learning about this anatomy from classical textbooks requires considerable intellectual gymnastics to deal with this spatially complex structure. We present a new way for teaching this anatomy with high resolution 3D reconstructions using X‐ray micro‐tomography.

Materials and Methods

10 human petrous parts of temporal bones were fixed with Winckler's solution. CT images were acquired with a CT scanner (Skyscan 1076, Bruker, Kontich, Belgium). The camera was continuously rotated by 180° and the following parameters were used for acquisition: 50 kV, 0.5 mm Al filter, 200 A source current, 35 m isotropic resolution, 180 ms exposure time, and 0.7° rotation step. The projections were reconstructed with a filtered backprojection algorithm in Skyscan software (NRecon, Skyscan). Reconstructed images were analyzed with ORS Visual software.(v1.5, Montreal), which allowed multiple display and post‐processing possibilities (3D VR; orthogonal/curved multiplanar reformation).

Results and Discussion

Three‐dimensional and multiplanar studies of the temporal bone morphology were possible with appropriate thresholding. The whole components of the middle ear (ossicles, tympanic cavity bony reliefs), the inner ear (cochlea, modiolus, cochlear canal, vestibule and semicircular canals) and their respective anatomic relationships are clearly visible. This method seems very effective in teaching this complex anatomy.

Conclusion

3D reconstructions using high resolution X‐ray micro‐tomography aid comprehension of temporal bone complex anatomy. This precise knowledge will help residents to better understand otological surgery.
From The Novice To The Experienced Ultrasound User: 7D Interactive Software Is Your Personal Tutor
Authors: Blandford Ian , Benninger Brion
Published: November 2023

INTRODUCTION

Ultrasound has become a very diverse tool in healthcare today. Ultrasound systems have improved significantly during the past decade. Unfortunately, the demand to learn is greater than available training. The objective of this study was to investigate if an ultrasound education software program could be successfully used by novice healthcare profession students.

METHODS

Literature search was conducted regarding interactive ultrasound educational software applications. Infrastructure of 7D Imaging educational ultrasound software includes text, imaging (static and video), testing integration, and clinical applications. Twenty students (10 baccalaureate (BS), 10 medical (MS)) involved in an anatomy course were exposed to the 7D Imaging ultrasound education software. Each student was asked to navigate through the program to aid them in conducting an ultrasound exam on the supraspinatus tendon, Morrison's pouch, and the femoral triangle. Two weeks later, we examined the students conducting ultrasound imaging and identify structures using Fukuda Denshi UF‐760AG ultrasound machine and Sonivate Soniceye finger probes.

Literature search revealed no interactive ultrasound education software programs. However, a plethora of random, informative demonstration videos exist, but lack comprehensiveness and testing ability. Of the thirty structures to be identified, the BS students successfully demonstrated and identified 25 out of 30 structures, and the MS 27 out of 30. Ultrasound continues to be used in virtually all disciplines of medicine. The vast majority of experts practice their skills with patients and have virtually no time to train novices. Therefore, there is a need for efficient interactive training software which allows self‐assessment.

CONCLUSION

This study revealed that the 7D Imaging interactive educational software could be successfully integrated as part of an anatomy course.3
The Effect of Focused Active Learning Strategies on Student Engagement with Respect to Socioeconomic Class
Authors: Birk Tirzah , Lee Valerie
Published: November 2023

Anatomy and Physiology are gateway courses for students to enter allied health professions. One of the primary ways that students originating from a low socioeconomic class are able to rise to the middle class is through entry into these allied health fields (Hlinka et al 2015). At the same time, those same students of low social and cultural capital are least likely to persist in higher education (Aragon 2000) (Wells et al 2011) (Wells 2008) (Smith et al 2015). The objective of this study was to determine if students of low social and cultural capital will report a higher sense of engagement, satisfaction, perception of group work, and retention of knowledge in introductory Anatomy and Physiology as well as grit, social‐belonging, perceptions of the school's culture and peer‐interactions compared to those of high social and cultural capital when focused active learning is increased. Data were collected by pre and post class surveys of students in a second semester introductory Anatomy and Physiology course using focused active learning exercises in lecture. Control surveys were sent to students in the same course in previous semesters with no active learning exercises during lecture. Pilot study data (n=14) revealed student preference for active learning over traditional lecture and general satisfaction and engagement in the active learning activities. Additionally, lower achieving students performed slightly better on active learning‐specific exam questions compared to higher achieving students (p>0.05). Preliminary data suggest that students of low social and cultural capital have increased perceptions of learning, and an increased sense of belonging due to increased peer and faculty interactions and practical applications inherent to active learning.
Microbiota‐Derived Tryptophan Metabolites Activate Aryl Hydrocarbon Receptor and Induce IL‐10 Receptor Expression in Intestinal Epithelia
Authors: Alexeev Erica E. , Lanis Jordi M. , Schwisow Kayla D. , Kominsky Douglas J. , Colgan Sean P.
Published: November 2023

Endogenous tryptophan metabolites play an important role in gut immune homeostasis, yet the role of tryptophan metabolites from host microbiota have not been extensively explored. Indole‐3‐aldehyde (IAld), a tryptophan metabolite produced by anaerobic intestinal bacteria, is known for its intercellular signal activity and serves as an aryl hydrocarbon receptor (AHR) ligand. The physiological function of AHR includes roles in immune regulation and mucosal barrier function. Further, IL‐10 is an anti‐inflammatory cytokine that inhibits production of numerous pro‐inflammatory mediators in various cell types, including intestinal epithelial cells (IECs). This cytokine functions through binding to the IL‐10 receptor alpha subunit (IL‐10R1). An unbiased metabolomic profile of whole colon tissue from healthy and colitic mice identified significant decreases in several microbial‐derived indole derivatives (e.g. indoleacetate, indolepropionate, 5‐OH‐indoleacetate). Guided by these results, we hypothesized that indole‐containing tryptophan metabolites regulate epithelial IL‐10R1 expression. Human intestinal epithelial cells were exposed to IAld and examined for expression of IL‐10R1. These results revealed a prominent induction of IL‐10R1 mRNA and protein expression following IAld treatment. Such results were explained, at least in part, by the activation of AHR by indole derivatives. Indeed, IL‐10R1 expression was diminished when IAld was administered to AHR nuclear translocator (ARNT) knockdown cells. Likewise, the AHR inhibitor, CH‐223191, significantly reduced IL‐10R1 expression in cells exposed to IAld. Together, these results provide strong evidence for the role of tryptophan‐indole derivatives in anti‐inflammatory pathways mediated by IL‐10 signaling in IECs. The discovery of tryptophan metabolites as AHR ligands provides new insight to our understanding of the host‐microbial axis during inflammatory processes. These studies also identify possible avenues for incorporating the indole‐AHR pathway as novel therapies for mucosal disease.
Markers of Hypoxia and Correlation with Disease Severity in Inflammatory Bowel Disease
Authors: Schwisow Kayla Danae , Colson Steven , Gerich Mark , Lovell Mark , Allshouse Amanda , Yeckes Alyson , Kessler Brittelle , Keely Simon , Hoffenberg Edward , de Zoeten Edwin , Colgan Sean
Published: November 2023

Ongoing inflammation results in a significant shift in tissue metabolism. Previous studies indicate that inflammation and hypoxia occur concomitantly. Here, we sought to determine whether circulating and tissue indicators of hypoxia could serve as surrogate indicators of disease severity in patients with inflammatory bowel disease (IBD). Serum and colon biopsies were obtained at a tertiary care center from 24 pediatric subjects with active IBD colitis and 26 non‐colitis controls. Serum was also collected from adult subjects: 10 with active IBD, 10 IBD patients with inactive disease, and 19 non‐colitis controls. Disease severity for the pediatric samples was graded by histopathology and inflammatory cytokine profile. Erythropoietin (EPO), vascular endothelial growth factor (VEGF), and macrophage inflammatory protein 3α (MIP3α) were measured in serum and tissue biopsies and correlated with disease severity. An initial screen of serum hypoxia markers revealed that children with colitis had higher circulating levels of EPO, VEGF and MIP3α than non‐colitis controls (all p<0.05). In parallel, analysis of colon tissue MIP3α RNA and protein revealed increased expression in children with IBD colitis compared to controls (p<0.05 and p<0.01 for RNA and protein, respectively). All three serum markers significantly differed across histology grade (VEGF p<0.01, EPO p<0.001, MIP3α p<0.001) and disease severity correlated with tissue IL‐6, IL‐8, and TNFα protein levels. In adult subjects, the same trends were seen in the circulating levels of EPO, VEGF, and MIP3α, with significant increases in EPO and MIP3α in active IBD patients compared to non‐colitis controls (p>0.0005 and p>0.05 for EPO and MIP3α, respectively). These results reveal that markers of hypoxia strongly correlate with histologically‐graded colitis as well as tissue inflammatory cytokine levels. Peripheral blood hypoxia markers may be useful surrogates to assess colon inflammation and could be utilized as potential indicators of disease activity for IBD.

Support or Funding Information

This work was supported by National Institutes of Health grants RO1‐HL60569, T32 DK067009, and UL1 RR025780 and by support from the Crohn's and Colitis Foundation of America.