bluuebunny/crossref_metadata_2025 huggingface dataset:
Semisynthetic Natural Products for Antibacterial Drug Discovery
Authors: Lee Richard
Published: November 2023

The pressing need for new treatments for multi‐drug resistant bacterial infections including MDR and XDR tuberculosis has led us to re‐explore the use of natural product antibiotics not normally used to treat target infections, especially those with unique mechanisms of action. Being the product of combinatorial evolution natural products have many advantages over purely synthetic antibacterial leads, but also some notable disadvantages often in regard to spectrum and human metabolism. This presentation will go over our efforts to advance under‐explored antibiotic scaffolds with semi‐synthetic approaches to tackle these disadvantages. Using spectinomycin as an example, synthetic modifications have been applied to overcome native bacterial efflux leading to potent leads with better than predicted in vivo efficacy in complex infection models. These observations have led us to examine active uptake and slow‐growing pathogen targeting hypotheses that are hard wired into many natural products, which are very hard to replicate by direct purely synthetic de novo antibacterial drug design.

Support or Funding Information

This work was supported by research grant R01AI090810 by the National Institutes of Health and the American Lebanese Syrian Associated Charities (ALSAC)
Regulation of Yeast Pah1 Phosphatidate Phosphatase in Lipid Synthesis
Authors: Carman George M.
Published: November 2023

In the yeast Saccharomyces cerevisiae, the synthesis of phospholipids in the exponential phase of growth occurs at the expense of the storage lipid triacylglycerol. As cells progress into the stationary phase, the synthesis of triacylglycerol occurs at the expense of phospholipids. Pah1 phosphatidate phosphatase plays an important role in this metabolism; the enzyme produces the diacylglycerol needed for the synthesis of triacylglycerol and simultaneously controls the level of phosphatidate for the synthesis of phospholipids. Loss of Pah1 results in severe phenotypes that include aberrant nuclear/endoplasmic reticulum membrane expansion, cell wall fragility, reduction in lipid droplets, vacuole fragmentation, fatty acid‐induced lipotoxicity, susceptibility to oxidative stress, and loss of chronological life span. PAH1 expression is induced throughout growth and the induction in the stationary phase is stimulated by inositol supplementation. Pah1 phosphatidate phosphatase activity is modulated by phospholipids, sphingoid bases, nucleotides, and through the phosphorylation and dephosphorylation of the enzyme, as well as by proteasome‐mediated degradation. The proteasomal degradation of Pah1, which occurs by ubiquitin‐dependent and ‐independent mechanisms, is governed by the phosphorylation/dephosphorylation of the enzyme.

Support or Funding Information

Supported by National Institutes of Health grants R37‐GM028140‐33 and R01‐GM050679‐22.
Nearer to nature: design and optimization of artificial enzymes
Authors: Hilvert Donald
Published: November 2023

Protein design is a challenging problem. We do not fully understand the rules of protein folding, and our knowledge of structure‐function relationships in these macromolecules is at best incomplete. Nature has solved the problem of protein design through the mechanism of Darwinian evolution. From primitive precursors, recursive cycles of mutation, selection and amplification of molecules with favorable traits have given rise to all of the many thousands of gene products in every one of our cells. An analogous process of natural selection can be profitably exploited in silico and in the laboratory on a human time scale to create, characterize and optimize artificial catalysts for tasks unimagined by Nature. Recent progress in combining computational and evolutionary approaches for enzyme design will be discussed, together with insights into enzyme function gained from studies of the engineered catalysts.
Posttranslational Modifications In Bacteria‐Host Interaction
Authors: Shao Feng
Published: November 2023

Many bacterial pathogens use a type III secretion system to inject virulent effector proteins into host cells. These effectors often use sophisticated biochemical strategies to manipulate host signal transduction pathway. I will present our work on four bacterial effector families, each of which defines a novel enzymatic posttranslational modification, and their functions in infection. The OspF effector from Shigella flexneri employs a MAPK phosphothreonine lyase activity to block host MAPK signaling and IL‐8 production. The CHBP/Cif family of effectors from Burkholderia pseudomallei and Enteropathogenic E. coli (EPEC) catalyze deamidation of Gln‐40 in ubiquitin and NEDD8, thereby inactivating host ubiquitin pathway and many related cellular processes. NleE from EPEC and Salmonella typhimurium carries out a novel cysteine methylation modification on key ubiquitin‐chain sensory proteins in host NF‐kB pathway, resulting in blocking of NF‐kB‐mediated inflammatory response during infection. The NleB effector targets host death domain‐containing proteins in the death receptor pathway for arginine GlcNAcylation and this unique modification is critical for EPEC replication in the colons of infected mice. On the host side, the inflammasome complex that activates inflammatory caspases is believed to be important for macrophage innate immune defense against bacterial infection. I will also discuss our recent identification and characterization of several cytosolic receptors for important bacterial molecules including LPS, flagellin and also Rho‐modifying effectors/toxins as well as the molecular mechanism for pyroptotic cell downstream of inflammatory caspases activation.

Support or Funding Information

The research was supported by the China National Science Foundation, China Ministry of Science and Technology, Beijing Government and the Howard Hughes Medical Institute.
Lycopene Inhibits Amyloid‐β25–35‐Induced Apoptosis by Decreasing Intracellular and Mitochondrial ROS Levels in Human Neuronal SH‐SY5Y Cells
Authors: HWANG SINWOO , Lim Joo Weon , Kim Hyeyoung
Published: November 2023

Alzheimer's disease (AD) is a major neurodegenerative disorder which is characterized by amyloid‐beta (Aβ) plaque and neurofibrillary tangles. The accumulation of Aβ in the brain is a crucial feature of AD, which causes cell death in AD by evoking a cascade of oxidative damage to neuronal cells. Lycopene, a carotenoid family, has been recognized as an antioxidant to protect oxidative stress‐induced cell damage in several cell lines.

In this study, we investigated the effect of lycopene on Aβ25–35‐induced cytotoxicity in human neuronal SH‐SY5Y cells. SH‐SY5Y cells were widely used a typical model for AD.

Cell viability was measured by MTT assay, and apoptotic evidence (bax, bcl‐2, and caspase‐3) was assessed by Western blotting.

Our results showed that exposure of cells to Aβ25–35 induces apoptotic cell death by increasing Bax/Bcl‐2 ratio and caspase‐3 activation. In addition, Aβ25–35 increased levels of intracellular reactive oxygen species (ROS) and mitochondrial ROS. Lycopene significantly suppressed viability loss and decreased apoptotic indices (increased Bax/Bcl‐2 ratio and caspase‐3 activation) in Aβ25–35 treated cells. Lycopene reduced the levels of intracellular and mitochondrial ROS in the cells, which was evoked by exposure to Aβ25–35.

These results suggest that lycopene attenuates the Aβ25–35‐induced apoptosis by decreasing intracellular and mitochondrial ROS levels in neuronal cells. Lycopene supplementation may be beneficial for preventing neuronal cell death of the patients with Alzheimer's disease.
Broccoli (Brassica oleracea) Extract Combats Streptozotocin‐Induced Diabetes and Oxidative Stress in Rats
Authors: Suresh Sithara , Waly Mostafa I , Guizani Nejib , Rahman Mohammad Shafiur
Published: November 2023

Background

The primary prevention of type 2 diabetes relies also on lifestyle changes, including dietary habits. The antidiabetic effects of several foods and dietary supplements in both animal models and in humans have been explored. It was found that beneficial effects are mainly dependent on antioxidant and hypoglycemic properties. Broccoli (Brassica oleracea) exerts beneficial effects in several diseases including type 2 diabetes, yet the mechanism was not elucidated.

Objective

This study aimed to evaluate the hypoglycemic and antioxidant potential properties of broccoli extract in diabetic rats.

Methods

Twenty male Sprague Dawley rats, weighting 250–300 grams, were allocated as into 4 groups (5 rats/group): non‐diabetic control that received chow diet; non‐diabetic control group that received chow diet plus oral feeding of broccoli extract; diabetic group that received chow diet; and diabetic group that received chow diet plus oral feeding of broccoli extract. Streptozotocin (STZ) drug was used as a diabetogenic agent in a single intraperitoneal injection dose of 60 gm/kg body weight, and the blood glucose level for each rat was measured twice a week. After 8 weeks, all animals were overnight fasted and sacrificed; serum was collected for measurement of glucose, and pancreatic tissues were homogenized for biochemical assessment of antioxidant enzymes (glutathione peroxidase and glutathione reductase), and oxidative stress markers (glutathione, total antioxidant capacity, lipid peroxidation, and hydrogen peroxide).

Results

Diabetic rats treated with broccoli extract had shown a significant protective effect against STZ‐induced hyperglycemia and oxidative stress as compared with the control group.

Conclusion

Our results provide evidence that broccoli extract possessed potential health benefits in controlling STZ‐induced diabetes in rats. The used in‐vivo model provides insight for the capacity of broccoli extract to be used for phytoremediation purposes in the context of human type 2 diabetes.
Comparison of Two Administration Forms of a Highly Viscous Fibre Blend on Appetite and Glycemic Response in Healthy Individuals
Authors: Au‐Yeung Fei , Jovanovski Elena , Ho Hoang , Zurbau Andreea , Jenkins Alexandra L , Vuksan Vladimir
Published: November 2023

Objective

The effect of viscous fibres on appetite and food intake have been equivocal, limited by the capacity of gel‐formation within the gastrointestinal tract. Differences in fibre administration form many determine the physiological effects of viscous fibres. Using a highly viscous fibre blend (VFB) of two complimentary fibres, konjac glucomannan and xanthan gum, we investigated the acute effects of pre‐hydrating VFB into a solid gel compared to the powdered form on appetite and glycemic response.

Methods

Eighteen (18) healthy participants completed the study (F: 11/M: 7; Age: 28.6 ± 3.2 years; BMI: 23.3 ± 0.7 kg/m2) and 15 provided blood samples. In an acute, randomized, double‐blinded, cross‐over design, participants consumed 1 of 5 test meals on 5 separate occasions with > 2 day washout period. The test meals were: 50 g available carbohydrates in bread with 5 g VFB as a powder (50Pow), 5 g VFB as a pre‐hydrated gel (50Gel), or a fibre‐free control (50Con), and 25 g available carbohydrates in bread with 25 g available carbohydrates in the gel dessert with 5 g VFB pre‐hydrated into the gel (25Gel) or a fibre‐free control (25Con). All meals consisted of bread, margarine, and a gel dessert which totaled 50 g of available carbohydrates. A metabolically controlled lunch was consumed 180 min later. Satiety was recorded using visual analogue scales.

Results

The incremental area under the curve (iAUC) after 120 min for blood glucose was lower after 50Pow (mean difference: −40 %, p < 0.001) and 50Gel (mean difference: −23 %, p = 0.049) vs. 50Con. No differences were observed between 50Pow and 50Gel (p = 0.16). Additionally, 50Gel increased satiety after 120 min vs. 50Pow (MD: +26 %, p=0.035) and 50Con (mean difference: +28 %, p = 0.025). The iAUC after 120 min for blood glucose was lower in 25Gel vs. 25Con (mean difference: −28 %, p = 0.006), but no significant differences were observed in satiety.

Conclusions

These findings suggest that VFB may improve the postprandial glycemic response irrespective of administration mode, but consumption in a pre‐hydrated gel may confer additional benefits from improved satiety.

Clinicaltrials.gov Identifier: NCT01657058
Predicting Initial Lipid Release from Masticated Tree Nuts Using Mathematical Modelling
Authors: Grassby Terri , Shen Youcong , Wagner Lily , Beckett Sarah , Hall Wendy , Berry Sarah
Published: November 2023

Introduction

The proportion of lipid released during mastication of nuts is strongly influenced by particle size, due to natural encapsulation of the lipid by the walls of intact cells. The cell walls may act as barriers to digestion and may partially explain why nuts have reduced metabolizable energy versus the energy content predicted by Atwater factors. The lipid released from masticated nuts can be calculated using a mathematical model which has the cell diameter and the particle size distribution (PSD) of the bolus as variables. This study measured the cell size and PSD of four tree nuts (raw cashews, raw walnuts, roasted pistachios and raw Brazil nuts) to predict the lipid released due to their mastication.

Objectives

To predict the proportion of lipid released from masticated tree nuts, using measurements of cell size and PSD of masticated tree nuts (including cashews, walnuts, pistachios and Brazil nuts). To compare the results for these nuts to those already published for almonds.

Method/Design

Transverse and longitudinal sections were cut from each nut (including almonds) and micrographs processed using image analysis software to calculate the average cell diameter. Two randomized, un‐blinded, cross‐over trials were conducted. In each trial, 10 healthy volunteers attended two sessions, at which they chewed eight samples of a randomly allocated nut. For determination of PSDs, the expectorated boluses were sieved (2 boluses) or analyzed by laser diffraction (2 boluses). Initial lipid release was then predicted using the mathematical model.

Results

The cashew cells (34.3 μm) were smaller than the almond (45.1 μm), walnut (49.4 μm), pistachio (53.1 μm), and Brazil nut cells (60.8 μm). Laser diffraction showed that masticated nut boluses had median particle sizes (± SEM) which were smaller (cashews, 178 ± 12 μm; walnuts, 179 ± 8 μm; pistachios, 123 ± 10 μm; Brazil nuts, 145 ± 8 μm) than that for almonds measured previously (550 ± 18 μm). This results in higher predicted lipid release, calculated from the mathematical model, (mean, range) for cashews (12.3%, 8.7–16.3%), walnuts (14.5%, 12.0–18.0%), pistachios (11.0%, 8.7–13.0%) and Brazil nuts (14.4%, 11.9–17.3%) than for almonds (9.5%, 7.4–11.1%).

Conclusions

All of the five nuts had predicted lipid releases of less than 18%, which would be expected to attenuate postprandial lipemia and total nutrient availability relative to that expected due to their total lipid content. Due to their higher lipid content and predicted lipid release, walnuts and Brazil nuts (after mastication) are likely to release more fat on mastication than the other nuts tested.

This research was approved by the relevant NHS National Research Ethics Committee (14/LO/2286 and 15/WS/0104) and registered at clinicaltrials.gov (NCT02398084 and NCT02497768).

Support or Funding Information

This research was funded by King's College London.
EGCG Decreases Mortality in a Dose‐Dependent Fashion but Does Not Improve Cognition in Aged Mice
Authors: Pence Brandt D. , Bhattacharya Tushar K. , Rytych Jennifer L. , Park Pul , Allen Jacob M. , Sun Yi , McCusker Robert H. , Kelley Keith W. , Johnson Rodney W. , Rhodes Justin S. , Woods Jeffrey A.
Published: November 2023

We have previously shown that 40 days of a diet containing epigallocatechin gallate (EGCG) and beta‐alanine was not effective in improving either cognitive or muscle function in aged (18 month) mice (Gibbons et al. Behav Brain Res 2014, Pence et al. Appl Physiol Nutr Metab in press). However, diet reduced oxidative stress in the brain, and previous studies using longer‐term interventions have documented beneficial effects in cognitive function, especially with EGCG. Therefore, we investigated whether a different dose of EGCG could be more efficacious in altering cognition. Aged (21–25 mo) Balb/cByJ male mice underwent 9 weeks of feeding with EGCG at 0, 0.09, or 4.1 mg/g AIN‐93M diet (n=15/group), then underwent a battery of cognitive tests. EGCG feeding did not alter preference for novel versus familiar arm in the Y‐maze test (p=0.292) and did not affect learning in the active avoidance test (p=0.758). Additionally, EGCG did not affect preference for novel versus familiar mice in a social exploration test (p=0.167). EGCG did decrease mortality in a dose‐dependent fashion (p=0.05, logrank test), with 91% of high EGCG, 72% of low EGCG, and 55% of control mice surviving to the end of the study. In conclusion, EGCG may improve survival in aged mice but does not affect cognitive function.

Support or Funding Information

Funded by the Abbott‐UIUC Center for Nutrition, Learning, and Memory
Secular Trends in Regional Differences in Biomarkers of Dietary Exposure and Inflammation among American Adults: NHANES III, 1988–1994 to C‐NHANES 1999–2010
Authors: Kant Ashima K. , Graubard Barry I.
Published: November 2023

Regional disparities in health outcomes, including age‐adjusted mortality rates, are evident at several different levels of geographic aggregation in the US. The regional gradient of health outcomes generally reveal the South and Midwest regions to compare unfavorably with the West and Northeast regions. Moreover, regional gaps in health outcomes have persisted over time. We used data from several NHANES surveys to examine time trends in the association of census region of residence (Northeast, Midwest, West, and South) with serum concentrations of several nutritional biomarkers among American adults aged ≥20 years. The biomarkers available for this period included: serum (and RBC) folate and c‐reactive protein (CRP) (NHANES 1988–1994 to 2009–2010, n>40,000); α‐carotene, β‐carotene, β‐cryptoxanthin, lutein‐zeaxanthin, lycopene, and vitamin D (NHANES 1988–1994 to 2001–2006; n>26,500), vitamin E (NHANES 1988–1994, 1999–2001 and 2005–2006, n> 26,500); vitamin C (NHANES 1988–1994 and 2003–2006, n>21,900); and vitamin B‐6 (NHANES 2003–2008, n>12, 800). In multivariable regression models adjusted for covariates, the interactions of survey cycle and region were not significant, indicating similar secular trends among regions. Main effect of region was significant (P<0.01) for all biomarkers except serum vitamin B‐6. Mean serum concentrations of all examined biomarkers were lower but that of CRP were higher in the South census region of the country. Generally the observed regional gradients in biomarker concentrations were in the same direction as those reported for health outcomes.

Support or Funding Information

Funded by National Institute of Aging grant (AG046540) (AKK).
Nut Intake, Prospective Weight Change, and Obesity Risk: The Adventist Health Study‐2
Authors: EL‐AMARI SALEM S , LLOREN JAN IRENE , SABATE JOAN
Published: November 2023

BACKGROUND

Nuts are currently recommended for lower serum cholesterol and reduces the risk of heart diseases. However, since they are high in energy and fat, long term consumption of nuts may lead to weight gain.

OBJECTIVE

To prospectively investigate the possible relationship between habitual nut intake and body weight gain and overweight/obesity risk.

RESEARCH DESIGN AND METHODS

Using data from Adventist Health Study‐2, an ongoing large cohort study, an analytical sample of 41,845 participants (65.5% female) with an average age of 55 years was examined. This analysis relate changes, between baseline and an avaerage of 8 years of follow‐up, in self‐reported body weight to regular nut consumption. Consumption of total nuts, including tree nuts and peanuts, (in grams/week) was assessed at baseline with a validated food frequency questionnaire. Odds ratios, and their corresponding 95% CI, were calculated from modelling unconditional logistic regression accounting for relevant demographic and many potential lifestyle confounders.

RESULTS

7,553 subjects gained ≥ 5kg of body weight and 23,372 became overweight/obese during the follow‐up. Subjects at the high tertile of tree nut intake (median intake of 21 gms/wk), compared to those at the low tertile (median intake of 1.3 gms/wk), had lower risk of gaining ≥ 5 kg body weight (OR 0.92, 95% CI 0.86, 0.99, p 0.028). Further adjustment for baseline BMI turned these findings to non‐significant. No significant association was observed between the risk of gaining ≥5 kg body weight and both peanut and total nut intake. In addition, among lean subjects at baseline, participants at the high tertile of total and tree nut intake; compared to those at the low tertile of intake, had lower risk of overweight/obesity (OR 0.73, 95% CI 0.68, 0.78, p < 0.0001) and (OR 0.69, 95% CI 0.65,0.74, p < 0.0001) respectively. However, no significant association was observed for peanut intake. Finally, among obese subjects at baseline, no difference in odds of becoming lean was observed among different levels of nut consumption.

CONCLUSION

Our results have shown that long term consumption of nuts is not associated with body weight gain, and actually decreases the risk of overweight and obesity. These results are contrary to the public misconception that nuts are fattening.

Support or Funding Information

National Cancer Institute funded the original study (5R01 CA094594).
Dietary Carotenoid, Retinol, and Ascorbic Acid Intake and its Association with Breast Density in Young Women
Authors: Bodor J. Nicholas , Jung Seungyoun , Klifa Catherine , Van Horn Linda , Dorgan Joanne F.
Published: November 2023

Breast cancer is the leading cause of cancer in women and is the second leading cause of cancer death. A number of studies have investigated antioxidants and other dietary factors in their role in reducing breast cancer risk. In particular, much focus has been on the potential protective properties of carotenoids, retinol, and ascorbic acid, though findings are inconsistent. This study examined the intake of these nutrients in association with breast density, a known strong risk factor for developing breast malignancies. Data from a cohort of young women (mean age 27), who participated in the Dietary Intervention Study in Children – 2006 Follow‐Up Study (N = 172), were analyzed. Carotenoid, retinol, and ascorbic acid intake was assessed using multiple 24‐hour dietary recall data. Measures of absolute and percent dense breast volume were obtained using magnetic resonance imaging. Multivariable mixed effects models assessed the relationship between carotenoid, retinol, and ascorbic acid intake and breast density measures. All models accounted for race, smoking status, education, parity, duration of sex hormone use, childhood body mass index z‐score, percent body fat, and total energy intake. After adjustment, women in the highest quartile of total carotenoid consumption had an average percent dense breast volume of 15.8 %, compared to 20.3 % among those in the lowest quartile (p‐value < 0.01). A comparable trend was found for ascorbic acid, where women with the highest intake had a percent dense volume of 18.1 %, compared to 23.9 % in the lowest intake group (p‐value < 0.001). Similarly, greater retinol consumption was associated with decreased absolute breast dense volume (trend p‐value < 0.05). Findings from this study suggest that greater intakes of carotenoids, retinol, and ascorbic acid are associated with lower breast density in young women. Whether this may confer a lower risk for developing breast cancer requires further exploration.

Support or Funding Information

Funding provided by: R01 CA104670 (Dorgan, JF)
FOOD ITEMS THAT TRIGGERING A SYMPTOM IN FUNCTIONAL GASTROINTESTINAL DISORDERS
Authors: dikmen derya , Göktaş Zeynep , Demir Aslıhan , Öztürk Ömer , Aksoy Evrim Kahramanoğlu , Köklü Hayretdin , Tuna Yaşar , Kekilli Murat , Korkmaz Hüseyin , Yılmaz Bülent , Asıl Mehmet , Köklü Gülşah , Köklü Seyfettin
Published: November 2023

Introduction

Irrıtable bowel syndrome (IBS) and functional dyspepsia (FD) are the most common functional gastrointestinal disorders. IBS is an idiopathic disorder with wide range of symptoms like diarrhea, constipation and abdominal pain. FD is a disorder in gastroduodenal region and it consists of pain and burning sensation in epigastrium and feeling of early satiety and fullness. Pathophysiology of both disorders is not clear yet. Extrinsic factors including psychological factors and foods may affect the course of those chronic disorders.

Aim

In the current study we aimed to investigate what type of foods trigger the gastrointestinal symptoms in patients with IBS and FD.

Methods & Materials

Diagnosis of FD and IBS were based on Rome III criteria. 631 subjects (159 with IBS, 167 with FD, 170 with both IBS and FD and 135 healthy control subjects) participated in the study from six hospitals located in different regions of Turkey. All the subjects completed a questionnaire which included questions about nutritional habits, physical activity, sleeping habits, a short form of food frequency questionnaire. Anthropometric measures like weight and height were self‐reported.

Results

The most prevalent food items that trigger the symptoms in FD were; carbonated drinks (15.1%), hot spices (14.8%) and fried foods (14.1%). In IBS subject group carbonated drinks (16.1%), hot spices, (13.8%) and legumes especially beans (8.2 %) and fried foods (7.9 %) were the main food items that trigger the symptoms. In the IBS and FD group, carbonated drinks (14.2 %), hot spices (11.7 %) and fried foods (9.8 %) were also the main food items causing a symptom. Although the main symptom triggering food item was carbonated drinks for all the groups, 50 % of the subjects were not abstaining from such drinks.

Conclusion

Although IBS and FD have different symptoms, both disorders might be affected by similar food items.
Effect of Dietary Carboxymethyllysine on glucose and insulin sensitivity, cecal fatty acids and the plasma metabolome in mice fed the Total Western Diet
Authors: Ward Robert Edwin , Xiao Siyu , Michalski Marie‐Caroline , Geloen Alain , Hintze Korry
Published: November 2023

Carboxymethyllysine (CML) is an oxidized product of lysine and a reducing sugar that is formed in heated foods. Rodent studies indicate that dietary CML is proinflammatory via interaction with the advanced glycation endproduct receptor (RAGE), and that dietary CML promotes insulin resistance. Moreover, previous in vitro studies have shown that heat damaged proteins affect the composition and metabolic activity of the microbiome, which may have systemic effects. Therefore, we hypothesized that increasing levels of CML on a model Western Diet background would decrease insulin sensitivity and possibly modify the metabolic activity of the microbiome. To test this hypothesis, we formulated versions of the Total Western Diet with increasing levels of CML. This was achieved by heating the sugar (glucose and fructose) along with the protein (casein). Subsequently, the protein/sugar mix was added to the diet at three levels. The result is that three diets were produced with <2, 8 and 16 ug CML per gram of diet. Mice (n=10/gp) were fed diets for 8 weeks. An oral glucose tolerance test (OGTT) was performed at week 6 and an insulin tolerance test at week 7. After 8 weeks mice were sacrificed and tissues were collected and frozen in liquid nitrogen. There were no differences in food intake or weight gain. At week 6 mice fed the high CML diet had increased fasting glucose (p<0.05), and yet a lower area under the curve (AUC) in the OGTT (p<0.05). There was also a trend for mice fed the highest CML diet to be the most insulin sensitive (p=0.052). The only difference in plasma metabolites was an increase in proprionic acid in mice fed the high CML diet vs. the low one. Subsequent analysis of the short chain fatty acids in the cecal contents indicate a dose‐dependent increase in proprionic acid as a function of dietary CML. Collectively, these data suggest that glycated protein may increase the production of proprionic acid in the gut as well as systemically, the significance of which is unclear.
Lactose Intolerance, calcium, vitamin D and parathyroid hormone levels in type 2 diabetic patients
Authors: Rana SatyaVati , Malik Aastha , Bhadada Sanjay K , Sachdeva Naresh , Morya Rajesh Kumar , Sharma Gaurav
Published: November 2023

Background

Type 2 diabetes mellitus is a chronic metabolic disorder. Common gastrointestinal symptoms in type 2 diabetic patients are nausea, vomiting, flatulence, constipation and/ or diarrhea. Reason for these symptoms may be lactose intolerance. Vitamin D, calcium and parathyroid hormone further regulate the bone mineralization in our body.

Aim

The present study was planned to measure lactose intolerance, levels of vitamin D, calcium, and parathyroid hormone in type 2 diabetic patients.

Material and Methods

150 patients and 150 age and sex matched healthy controls attending Endocrinology Clinic in PGI, Chandigarh were enrolled. Lactose intolerance was measured using non‐invasive lactose breath test. 25‐hydroxyvitamin D3 and Parathyroid hormone were measured in plasma using immunoassay. Plasma calcium was measured using auto analyzer. T score was recorded from DXA scan for bone mineral density measurement.

Results

Lactose intolerance was observed in 89/150 (59.3%) diabetic patients and 63/150 (42%) in controls. Levels of vitamin D3, calcium and parathyroid hormone were significantly less in diabetic patients as compared to controls. Furthermore, levels of plasma vitamin D3, calcium and parathyroid hormone were more decreased in lactose intolerant diabetic patients than lactose tolerant patients. 67% of diabetic patients suffered from osteoporosis and 20% of controls. 80% diabetic patients and 16% controls with osteoporosis suffered from lactose intolerance.

Conclusion

From this study we can conclude that measurement of lactose intolerance using non‐invasive lactose breath test is suggested for type 2 diabetic patients so that deficiency of vitamin D and calcium can be taken care off.

Support or Funding Information

Indian Council of Medical Research, New Delhi, India
Lipids and Cholesterol‐Lowering Activity of Red Cabbage Microgreens
Authors: Jiang Xiaojing , Huang Haiqiu , Xiao Zhenlei , Yu Lu , Pham Quynhchi , Yu Liangli (Lucy) , Luo Yaguang (Sunny) , Wang Thomas T.Y.
Published: November 2023

Cardiovascular disease (CVD) is the leading cause of death in the United States and diet‐induced hypercholesterolemia is a major risk factor. Population, as well as experimental evidence, support that the consumption of a variety of vegetables can lower the risk of CVD through modulation of hypercholesterolemia. Microgreens are tiny edible greensharvested from vegetables and herbs at 7–14 days after germination, and are reported to possess a higher density of nutrients when compared to their mature counterparts. However, little is known about the health promoting efficacies of microgreens in terms of their effect on lipid and cholesterol levels. The present study used a rodent diet‐induced obesity model to address this question. C57B/6NCr mice (n=62, male, five week) were randomly assigned to six groups: 1) low fat diet (with 10% calories from fat); 2) high fat diet (with 45% calories from fat); 3) low fat diet+1.95%red cabbage microgreens; 4) low fat diet+1.95% mature red cabbage; 5) high fat diet+1.95% red cabbage microgreens; 6)high fat diet+1.95% mature red cabbage. The animals were on their respective diets for eight weeks. We found supplementing microgreens attenuated high fat diet induced weight gain after 8 weeks (17%). Moreover, we found microgreens significantly lowered circulating LDL levels (34%), hepatic triglycerides (23%)and cholesterol ester (65.5%) levels in animals fed a high fat diet. The effects of microgreens on hepatic triglycerides and cholesterol ester appeared to coincide with inhibition in triglycerides synthesis enzymes diacylglycerol O‐acyltransferase 1 (DGAT1), glycerol‐3‐phosphate acyltransferase (GPAM), cholesterolester synthesis enzymes acetyl‐CoA acetyltransferase 3 (ACAT3) and lecithin‐cholesterol acyltransferase (LCAT). However, no changes in expression in the genes related to cholesterol metabolism including sterol regulatory element binding transcription factor 2 (SREBP2), 3‐hydroxy‐3‐methylglutaryl‐CoA reductase (HMGCR), acyl‐CoA oxidase (ACOX), cytochrome P450, family 7, subfamily A, polypeptide 1 (CYP7A1) were observed. In summary, our data support dietary consumption of red cabbage microgreens modulates cholesterol metabolism, weight gain, and may protect against hypercholesterolemia and CVD.

Support or Funding Information

Thomas T. Y. Wang Address:Room 132, 10300Baltimore Ave, Bldg. 307‐C, BARC‐EAST, Beltsville, MD, 20705Phone number: 301‐504‐8459 Fax:301‐504‐9062Email address: tom.wang@ars.usda.gov
Consistent Weight Gain Tracking is Associated with a Reduction in Gestational Weight Gain in High Income Women
Authors: Olson Christine M. , Graham Meredith L. , Strawderman Myla S.
Published: November 2023

Objective

To determine the importance of using the weight gain tracker in a multi‐feature integrated mobile phone and online healthy lifestyle intervention designed to prevent excessive gestational weight gain (GWG).

Methods

A 3‐arm randomized controlled trial with pregnant women in two early pregnancy body mass index (BMI) groups: normal and overweight plus obese class 1; and two income groups: low (Medicaid‐eligible) and high household incomes was conducted to evaluate the effectiveness of the integrated mobile phone and online intervention. Two arms received intervention during pregnancy. Data were collected through online surveys, a medical record audit, and weight tracking was automatically logged by the project website. Data were analysed using both modified Poisson and linear regression models controlling for sociodemographic and timing of weight measurement covariates. Three gestational weight gain outcomes were examined: prevalence of excessive total GWG; prevalence of excessive weekly GWG from 20 weeks gestation until delivery; and total GWG in kg. Consistent weight gain tracking was defined as at least one weight entry in each 45 day time interval of enrolment in the study.

Results

A racially and ethnically diverse sample of 1,335 18 to 35 year‐old healthy pregnant women was recruited into the trial through community‐based practices and clinics and online enrolment. Among higher income women, 49% of women with normal BMI and 27% of overweight plus obese BMI women were consistent weight gain trackers. Tracking was less common among lower income women: 13% among normal BMI and 11% among overweight plus obese BMI. Among higher income normal BMI women, consistent weight gain tracking was significantly associated with all three outcomes: reduced risk of excessive total GWG (p = 0.03); reduced risk of excessive weekly rate of GWG (p = 0.0009); and 1.73 kg less total GWG (p = 0.0002). Among overweight plus obese higher income women, consistent tracking was associated with near significant reductions in the two measures of excessive GWG (p = 0.07 and 0.09) and a significant reduction of 3 kg in total GWG (p <0.0001). Consistent weight gain tracking was not associated with any of the GWG outcomes in low income women.

Conclusions

Among pregnant women with household incomes above the Medicaid eligibility cut‐off, consistent weight gain tracking is an important feature of online and mobile interventions for reducing excessive GWG.

Support or Funding Information

Grant No. HL096760
The Role of p120‐catenin and PIK3CA in Migration in Head and Neck Squamous Cell Carcinoma
Authors: Kidacki Michal , Lehman Heather L , Welsh Patricia A , Warrick Joshua I , Stairs Douglas B
Published: November 2023

Over half a million people worldwide are diagnosed with head and neck squamous cell carcinoma (HNSCC). A third of the newly diagnosed patients won't survive the next five years. Additionally, approximately 15% of these patients already have distant metastases at the time of diagnosis. Therefore it is of profound interest to understand the mechanism driving HNSCC metastasis.

p120‐catenin (p120ctn) is a tumor suppressor that is lost in 19% of HNSCC. PIK3CA, a gene coding for the catalytic subunit of PI3‐kinase, is mutated in 21% (E545K and H1047R being the most common mutations) and overexpressed in an additional 20% of HNSCC. Both, p120ctn loss and PIK3CA mutations individually have been shown to play a role in metastasis in a number of epithelial cancer models. Considering the frequency at which both p120ctn and PIK3CA are dysregulated in HNSCC, it is very likely that these two changes may be found in the same patients, increasing a cancer's aggressiveness. Therefore, we hypothesize that cells harboring both p120ctn down‐regulation and PIK3CA mutations may represent a more malignant cell population than the cells with either of these changes alone.

The tumorigenic effects of p120ctn down‐regulation and PIK3CA mutations were tested using normal oral keratinocytes (NOK). NOK cell lines were generated by down‐regulating p120ctn alone or in conjunction with an overexpression of either of the PIK3CA mutants, E545K or H1047R. Migratory and invasive capabilities of these cell lines were assessed. Boyden chamber migration assays showed that either p120ctn knockdown or each of the PIK3CA mutations alone increased cell migration 2.5 to 3‐fold compared to the control. However, when p120ctn knockdown was coupled with either the E545K or H1047R PIK3CA mutation, the cell migration increased 4‐fold. A similar increase was observed for the invasive capabilities of these cells.

Signaling pathways were interrogated to identify the mechanism(s) responsible for the observed increase in cell migration. Both phospho‐STAT3 (Y705) and active, GTP‐bound RhoA were found to be increased in NOKs with p120ctn down‐regulation or PIK3CA mutations alone. Even greater increases were detected in cell lines containing both p120ctn down‐regulation and PIK3CA mutations. We are currently testing whether these increases in phospho‐STAT3 or active RhoA are responsible for the observed increase in cell migration and whether crosstalk between these signaling pathways exists. In conclusion, both p120ctn down‐regulation and PIK3CA mutations act cooperatively to increase migration/invasion, possibly via the STAT3 or RhoA pathway.
Regulation of Cardiac Autophagic Flux In Vivo by the Ubiquitin Ligase Muscle Ring Finger‐1 (MuRF1)
Authors: Parry Traci L. , Willis Monte S.
Published: November 2023

The ubiquitin‐proteasome and autophagy systems are comdplementary protein degradation pathways that play a critical role in protein quality control in cardiomyocytes. Our laboratory studies the muscle specific ubiquitin ligase MuRF1 and have identified that mice with cardiac‐specific increased MuRF1 expression is cardioprotective in cardiac I/R injury, while also more susceptible to heart failure when challenged with pressure overload‐induced cardiac hypertrophy. Since these phenotypes are consistent with increased autophagic flux, we tested the hypothesis that MuRF1 positively regulated cardiac autophagy. We first crossed GFP‐RFP‐LC3 Tg+ mice created to track autophagy with our MuRF1 Tg+ mice to create MuRF1/GFP‐RFP‐LC3 dual Tg+ mice. Using confocal analysis of early (dual green and red) and late (red) punta representing the autophagosome‐autolysosome transition, we identified that MuRF1 Tg+ mice had significantly increased cardiac autophagic flux. MuRF1 Tg+ hearts also had significantly increased LC3II levels by immunoblot post‐bafilomycin treatment indicative of increased autophagic flux. Conversely, MuRF1−/− hearts had significantly decreased LC3II levels by immunoblot analysis, consistent with decreased autophagic flux. We next hypothesized that MuRF1 supported autophagy by post‐translational modification of FoxO1 and FoxO3a, two major transcription factors previously described to regulate autophagy. Immunoblot analysis of the inactive p‐FoxO1 and p‐FoxO3a proteins in MuRF1Tg+ hearts demonstrated decreased p‐FoxO3a (p<0.05), while MuRF1−/− hearts had increased levels (p<0.05), consistent with MuRF1's support of Foxo1/3 activity. To determine if MuRF1 regulated FoxO activity transcriptionally, FoxO3a activity assays were performed by in vitro luciferase assays. Increasing MuRF1 levels were shown to enhance FoxO3a transcriptional activity (p<0.05), offering one potential mechanism by which MuRF1 supports autophagy in vivo. Lastly, we determined if MuRF1‐induced autophagy was cardioprotective in doxorubicin‐induced cardiomyocyte death, a model where increased autophagy has been reported to be protective. When MuRF1 expression was increased in the H9C2 cardiomyocyte‐derived cell line and challenged with 1 microM doxorubicin for 24 hours, a 66% reduction in Caspase 3/7 activity was seen compared to control cells, indicating that MuRF1's cardioprotection may be due, in part, to enhanced autophagy. These findings establish MuRF1 as the first ubiquitin ligase to regulate cardiac autophagy, which occurs via the FoxO3a transcription factor. Understanding MuRF1's regulation of autophagy provides insight for the development of therapies targeting autophagy in heart disease.
Nrf2 But Not Autophagy Activation Is Associated with Resistance to EGFR Inhibitor‐Induced Lung Tumor Cell Apoptosis
Authors: Li Yuan , Zhou Yan , Ni Hong‐Min , Zhong Hua , Ding Wen‐Xing
Published: November 2023

Objective

To understand the mechanism of the resistance of non‐small cell lung cancer (NSCLC) to epidermal growth factor receptor (EGFR) inhibitor treatment.

Background

NSCLC is one of the most common malignancies in the world, of which a group of patients have activating EGFR mutations. Icotinib and Gefitinib are two tyrosine kinase inhibitors (TKIs) that bind to the tyrosine kinase domain in EGFR and inhibit EGFR activity, which have been used to treat NSCLC. However, natural and acquired resistance significantly limits the efficacy of EGFR‐TKIs, and the mechanism of the resistance is not fully understood. Autophagy is a highly conserved, genetically programmed degradation pathway that removes damaged organelles and protein aggregates in response to various stimuli to maintain cellular homeostasis. Autophagy inhibits tumorigenesis but promotes tumor progression at later stages. Nuclear factor (erythroid‐derived 2)‐like 2 (Nrf2) is a transcription factor and regulates the expression of antioxidant genes. Activation of Nrf2 is a protective mechanism for normal cells under oxidative stress, and also for cancer cells from chemotherapeutic agents that facilitates cancer progression. Whether autophagy and Nrf2 activation contribute to the sensitivity of NSCLC to TKI treatment has been incompletely understood.

Methods

We compared the sensitivity of A549, an EGFR wild type cell line derived from lung carcinoma, and HCC827, an EGFR mutant cell line derived from lung adenocarcinoma, to TKI treatment, and determined the role of autophagy and Nrf2 activation in TKI‐mediated cell death.

Results

Icotinib and Gefitinib induced significant apoptosis in HCC827 cells in a dose‐dependent manner, while they did not cause apoptosis in A549 cells. TKI inhibitors induced autophagy flux and mTOR inhibition in HCC827 but not in A549 cells. Intriguingly, suppression of autophagy by chloroquine, a lysosomal inhibitor, did not affect TKI‐induced cell death. A549 cells had lower basal level of autophagy but higher basal level of Nrf2 activation than HCC827 cells. Brusatol, a Nrf2 inhibitor, did not affect autophagy in A549 or HCC827 cells, but significantly decreased the cell viability in A549 and HCC827 cells. However, Brusatol did not further sensitize A549 or HCC827 cells to TKI‐induced cell death.

Conclusion

A549 cells are more resistant to TKI‐induced apoptosis than HCC827 due to persistent activation of Nrf2. Although autophagy activity is higher in HCC827 cells than A549 cells, it is not essential for the sensitivity of HCC827 or A549 cells for TKI. Inhibition of Nrf2 reduces the cell viability of TKI resistant A549 cells although it does not show synergistic effects with TKI.

Support or Funding Information

NIH RO1 AA020518, NIH R01 DK 102142, P20 RR021940 (COBRE)
Neutrophil Extracellular Traps Drive Bleomycin‐Induced Lung Fibrosis by Regulating TGFβ1‐Dependent Interactions of Platelets and Macrophages
Authors: Riehl Dennis R. , Roewe Julian , Klebow Sabrina , Esmon Naomi L. , Eming Sabine , Colucci Giuseppe , Schäfer Katrin , Gunzer Matthias , Waisman Ari , Ward Peter A. , Esmon Charles T. , Bosmann Markus
Published: November 2023

Neutrophil extracellular traps (NETs) are extracellular histones and DNA expelled from polymorphonuclear neutrophils (PMNs). NETs are released during inflammation as an innate immune defense mechanism. NETs not only exert bactericidal activities but are toxic for host cells and thus promote tissue injury. In the past, many reports have defined the important roles of NETs during acute infections and acute inflammatory diseases. On the other hand, our current knowledge on the contribution of NETs during chronic inflammation, adaptive immunity and fibrotic diseases leaves much to be desired.

In this report, we uncover the immunologic networks initiated by NETs during the natural course of bleomycin‐induced lung fibrosis in mice. First, we designed a model of inducible transient genetic depletion of Ly6G+ PMNs by injection of diphtheria toxin in Ly6G‐Cre × iDTR mice. Depletion of Ly6G+ PMNs simultaneously with intra‐tracheal administration of bleomycin was sufficient to decrease the build‐up of extracellular matrix, T cells and cytokines (e.g. TGFβ1) in lungs after 4 weeks. Bleomycin mediated the release of citrullinated extracellular histones in broncho‐alveolar lavage fluids in dependency of PMNs with a surge after 2 days. Neutralization of extracellular histones by monoclonal anti‐histone H4 antibodies significantly reduced the expression and accumulation of extracellular matrix proteins and TGFβ1 during lung fibrosis. Platelets represented a major source (≥70%) of plasma TGFβ1 and purified extracellular histones activated platelets to release TGFβ1. Accordingly, bleomycin‐induced lung fibrosis was ameliorated in mice with platelet‐specific genetic deletion of TGFβ1 (PF4‐Cre × TGFb1flox/flox mice). TGFβ1 signaling required TGFβ receptor II and multiple down‐stream signaling pathways in macrophages. Mice with specific genetic ablation of the TGFβ receptor II in LysM+ macrophages (LysM‐Cre × TbRIIflox/flox mice) were also protected in bleomycin‐induced fibrosis. Finally, we found that histone‐induced TGFβ1 from platelets modulated the T cell tropic cytokine production in macrophages.

Our data suggest that NETs are an important factor for the pathogenesis of lung fibrosis. NETs may coordinate subsequent cytokine‐dependent interactions of platelets with innate and adaptive immune cells. In the future, pharmacologic blockade of NETs may be further evaluated as a potential treatment for chronic fibrotic diseases.

Support or Funding Information

This study was supported by the Federal Ministry of Education and Research (BMBF 01EO1503) and the Deutsche Forschungsgemeinschaft (BO3482/3‐1). The authors are responsible for the contents of this publication.