bluuebunny/crossref_metadata_2025 huggingface dataset:
Sugar‐sweetened beverages consumption is associated with BMI in Mexican adolescents. National Health and Nutrition Survey‐2006
Authors: Jiménez Alejandra , Flores Mario , Shamah Teresa
Published: June 2021

The objective was to evaluate the association between consumption of sugar‐sweetened beverages (SSBs) and body mass index (BMI) in Mexican adolescents.

Data from the National Health and Nutrition Survey‐2006 of Mexico were analyzed. Consumption of SSBs (i.e: sodas, fruit juices and sweet‐flavored drinks) was evaluated by means of a semi‐quantitative food frequency questionnaire. BMI was calculated as weight in kg/height in mt squared.

Information on socio‐demographic characteristics, sexual maturity, parents′ obesity, physical activity and inactivity (i.e: TV‐watching) during the week before the interview, was also obtained.

A total of 10,689 participants who represented 10,029,247 Mexican adolescents 10 to 19 y (mean age 13.8 ± 2.7 y) were evaluated. 50.4% were females. Mean time spent watching TV was 7.5 ± 5.5 h/week. Mean time of moderate and intense activity was 6.0 ± 4.0 h/week.

Mean BMI was 21.4 ± 4.4. The median consumption of SSBs was 1.08 (0.76,1.43) servings per day, after adjusting by energy intake, age, sex, physical activity, TV‐watching and socioeconomic level.

Multiple‐linear regression analysis showed that, for each portion of sodas consumed, a 0.20 ‐point increase in BMI was observed in males (p<0.05), after adjusting for confounders.

It was concluded that consumption of sodas is positively associated with BMI in Mexican male adolescents.

Supported by Ministery of Health, Mexico.
Effectiveness of legume consumption for facilitating weight loss: a randomized trial
Authors: McCrory Megan A , Lovejoy Jennifer C , Palmer Philip A , Eichelsdoerfer Petra E , Gehrke Malinda M , Kavanaugh Ian T , Buesing Scott A , Rose Teri L
Published: June 2021

Legumes are rich in factors which help promote satiety, including fiber, resistant starch and protein. Observational and multi‐meal experimental studies suggest that consuming legumes may help support healthy weight; however, randomized trials are lacking. We are conducting a 6‐wk randomized trial with 6‐wk follow‐up to test whether consuming legumes facilitates weight loss and improves chronic disease risk. After baseline measurements, subjects are advised to reduce daily energy intake (EI) by 30%. About half of the target EI is provided and consumption required, while the remainder of EI is self‐selected. Subjects are randomly assigned to 1 of 3 groups: DG: the US Dietary Guideline recommendation of 3 c/wk of legumes (0.5 c/d, 6 d/wk); DRI: the amount of legumes needed to meet the Dietary Reference Intake for fiber (women 1.8 c/d, men 2.7 c/d, 6 d/wk); or control: minimal legumes. To date, 28 subjects have enrolled (goal n=66). Mean±SEM weight loss thus far was: wk 3 (−1.7±0.3 kg), wk 6 (−2.7±0.7 kg), and wk 12 (−2.5±0.8 kg) (p 0.01 for each)). At wk 3, weight loss was greater in the DG and DRI groups (−2.3±0.4 kg; −2.4±0.7 kg) vs the control group (−0.9±0.4 kg)(n=19; p=0.046). At wk 6, weight loss also differed among groups (DG: −3.4±0.9 kg; DRI: −3.9±1.4 kg; control: 1.0±1.1 kg (n=15; p=0.11–0.12 control vs DG/DRI). Consuming at least 0.5 c/d of legumes may assist with weight loss and maintenance. [Funding: Pulse Canada]
Diacylglycerol consumption and blood lipids and adiposity in rats
Authors: DeIulio Emily , Precht Kristy , Zamudio Abigail , Nemoseck Tricia , Stocker Kelly , Kern Mark
Published: June 2021

Diacylglycerol (DAG) rich oil can promote lower adiposity and concentrations of serum triglycerides (TG) compared to triacylglycerol (TAG) rich fats. Most research, however, has assessed the influence of DAG fed at relatively high levels rather than the moderately low amounts evaluated here. Sixteen male weanling Sprague‐Dawley rats were equally divided into two groups. Rats consumed diets consisting of 21% fat by weight, mainly from coconut oil. DAG replaced 4% of the coconut oil for one group. After 30 days, serum total cholesterol (TC), HDL‐C, TG, glucose, and epididymal fat pad weights were assessed. Food intake (428 ± 46 vs. 457 ± 45 g), weight gain (184 ± 20 vs. 189 ± 15 g), food efficiency ratio (0.43 ± 0.05 vs. 0.41 ± 0.04), fat pad weights (2.740 ± 0.822 vs. 2.811 ± 0.488 g), TG (41.0 ± 20.0 vs. 51.3 ± 10.3 mg/dl), TC (78.7 ± 14.0 vs. 80.7 ± 18.8 mg/dl), and glucose (160.8 ± 31.1 vs. 169.8 ± 41.7 mg/dl) were similar (p>0.05) between the DAG and coconut oil fed rats, respectively. HDL‐C concentration (43.8 ± 6.0 vs. 56.5 ± 5.8 mg/dl) was lower for DAG‐fed rats (p<0.05). Results suggest that consuming a diet containing excess levels of saturated fat with moderately low amounts of DAG does not significantly improve lipids or reduce fat storage and may actually lower HDL‐C. Researchers should examine the minimum amount of DAG required to elicit positive responses and as a part of what type of diet the effects are most pronounced.
Assessment of the Docosahexanoic Fatty Acid Status in the Post‐Partum Period in Alexandria, Egypt
Authors: WALY MOSTAFA IBRAHIM‐AHMED , HABIB YOSR ABDEL‐AAL , SALEM EGLAL GHONEIM , DARWISH OLFAT ABDEL‐HAMID
Published: June 2021

Docosahexanoic acid (DHA) is a polyunsaturated fatty acid (PFA) of the omega‐3 series and is essential for the proper cognitive functions and normal development of nervous system. During pregnancy, the DHA requirement increase progressively as pregnancy advances and becomes the highest during the last trimester. Depletion of DHA as a result of inadequate supply or repeated pregnancy may induce neurological and psychological manifestations such as depression. This cross‐sectional study was conducted in Alexandria's maternal health centers and 250 women in the child bearing period were chosen for this study and the subjects were categorized into 4 groups, based on the number of pregnancies. Serum DHA measurement for the control group, who had no history of pregnancy, was 1.02 mg/mL, and we found that the serum DHA level for the other groups was decreased gradually with the increase in the number of pregnancies as follow, 0.51 mg/mL for the one pregnancy group, 0.082 mg/mL for the two pregnancies group, 0.073 for the three pregnancies group, and 0.042 mg/mL for the four pregnancies group. All subjects involved in this study had a low dietary intake of the rich sources of PFA. These findings suggest that the number of pregnancies is a stress factor contributing to the decreasing serum DHA level. Nutritional education and fortification of food with DHA should be considered to increase the dietary intake of the DHA to this population.
Regulation of arginine metabolism by dietary fatty acids. Involvement of PPAR‐alpha
Authors: Guelzim Najoua , Mariotti Francois , Lasserre Frederic , Mathe Veronique , Azzout Dalila , Pineau Thierry , Huneau Jean‐Francois , Hermier Dominique
Published: June 2021

Nitric oxide (NO) production depends on Arginine (Arg) competitive channelling towards urea (via Arginase) vs NO (via NO synthase, NOS). The regulation of these pathways by dietary PPARα ligands was explored in wild‐type (WT) and PPARα‐null (KO) male mice, fed diets rich in either saturated fatty acids (SFA) or 18:3 n‐3 (ALA). Metabolic markers of Arg metabolism were assayed in urine and plasma. Whole‐body NO production was assessed via the conversion of an IP bolus dose of [15N2‐guanido]‐arginine into urinary 15NO3 vs [15N2]‐urea. Enzymatic markers (protein and/or mRNA levels of Arginase and of endothelial and inducible NOS) were determined in liver. Diet effects in WT mice. In ALA‐fed mice, protein (Arginase) and mRNA (Arginase and both NOS) levels were half those in SFA‐fed mice. However, plasma urea and Arg, urine urea and nitrates, and whole‐body conversion of Arg into NO did not differ between diets. Effects of PPARα deficiency. In KO mice, Arginase protein and mRNA levels were half those in the WT groups, without effects on plasma and urinary metabolic markers, or on NOS expression. Besides, PPARα invalidation tended to reduce the dietary effects. However, the whole‐body conversion of Arg into NO tended to be lower (by 50%) in KO mice fed ALA than in those fed SFA. Conclusion. In this model, the nature of dietary fatty acids (SFA vs ALA) impacts Arg metabolic pathways, of which regulation might be mediated in part by PPARα.
In vitro effects of arginine or citrulline associated with a statine on nitrite oxide (NO) production in bovine aortic endothelial cells (BAECs)
Authors: Berthe Marie‐Clotilde , Bernard Mélisande , Darquy Sylviane , Couderc Rémy , Cynober Luc
Published: June 2021

Introduction Endothelial dysfunction occured very early in atherogenesis. It is due to decrease in NO production by endothelial cells leading to loss of vasodilation. NO is synthetised in endothelial cells by the endothelial nitric oxide synthase (eNOS) from arginine, with production of citrulline. Citrulline can be recycled in arginine, via arginino succinate synthase (ASS) and arginino succinate lyase (ASL). Moreover, statine increases eNOS activity and synthesis and therefore improves endothelial function.

Materials and methods NO was determined using a nitrate and nitrite fluorimetric assay.

Results

NO production was increased in the presence of simvastatine and arginine or citrulline. This increase was more important with arginine compared to citrulline at 1mM. Without any substrate in the medium, an increase in eNOS protein level was found. In the presence of arginine 1mM, the simvastatine effect on eNOS protein level was more pronounced than in the presence of arginine 0,5mM. The effect was found also in the presence of citrulline 1mM. The same results were observed with eNOS mRNA level. Finally, when cells were incubated with arginine 0,5mM, simvastatine lead to increase ASL and ASS mRNA expression.

Conclusion

The association of substrate with simvastatine induces an increase NO production, compared with arginine or citrulline or statine only, which is in agreement with the increase in BAECs of eNOS mRNA and protein levels.
Maximizing protein anabolism in young and aging muscle: a dose response to dietary protein ingestion
Authors: Symons T. Brock , Sheffield‐Moore Melinda , Wolfe Robert R. , Paddon‐Jones Douglas
Published: June 2021

We have recently shown that a single moderate serving of a protein‐rich food (113 g of lean beef; 30 g protein) acutely increases muscle protein synthesis by 51% in both young and elderly.

PURPOSE:

To quantify the age‐ and dose‐response to nutrient ingestion by measuring muscle protein synthesis following a single large serving of a protein‐rich food (340 g lean beef; 90 g protein; ~1.2g/kg/day for 70kg individual) in young (n=7; 29±3yrs) and elderly (n=7; 66±2yrs) individuals.

METHODS:

Venous blood and vastus lateralis muscle biopsy samples were obtained during a primed (2.0 μmol/kg) constant infusion (0.08 μ mol·kg−1min−1) of L‐ [ring‐13C6] phenylalanine. Mixed muscle fractional synthesis rate (FSR) was calculated during a 3 h post‐absorptive period and for 5 h following ingestion of 340 g of 90% lean ground beef.

RESULTS:

Mixed muscle FSR increased by approximately 46% in both the young (pre: 0.082±0.010; post: 0.119±0.012) and the elderly (pre: 0.086±0.008; post: 0.125±0.014 %/h) following ingestion of 340 g of lean beef. These data were similar to the response to ingestion of 113 g lean beef in the young (pre: 0.074±0.005; post: 0.113±0.005 %/h) and elderly (pre: 0.072±0.004; post: 0.108±0.006).

CONCLUSION:

A large serving of protein increases muscle protein synthesis by 46% in both young and elderly, but provides no additional anabolic stimulus above that afforded by a more moderately sized 113 g serving. These data have implications for the dose and distribution of dietary protein over the course of a day.
Glucotoxicity‐induced apoptosis and suppression of cell proliferation in pancreatic beta‐cells
Authors: Han Diana , Yang Byungho , Claycombe Kate J. , Yu Seong‐Woon , Kim Eun‐Kyoung
Published: June 2021

Diabetes is associated with an increased risk for a number of serious, sometimes life‐threatening complications. It is a loss of beta‐cell function and/or mass that eventually defines the disease. Glucose, the main regulator of insulin secretion and production, exerts negative effects on beta‐cell function when present in excessive amounts over a prolonged period. This chronic hyperglycemia is detrimental to pancreatic beta‐cells, causing impaired insulin secretion and playing an essential role in the regulation of beta‐cell turnover through a poorly known process, glucose toxicity (glucotoxicity). Our studies demonstrate that chronic exposure of hyperglycemia (30 mM glucose) to INS‐1 pancreatic beta‐cells causes an apoptotic programmed cell death, which coincides with induction of caspase‐3 activation and DNA fragmentation, but without a change in Bcl‐2 and Bcl‐xL protein levels. Our results also indicate that pancreatic beta‐cells undergo a reduced proliferation mediated by decreased p70S6K activation following hyperglycemia. Accordingly, our studies suggest that hyperglycemia induces caspase‐dependent pancreatic cell apoptosis and suppress cell proliferation, thereby leading to a reduction in cell viability.
The National Mindless Eating Challenge: Dietary Profiles Predict a Differential Effectiveness of Interventions:
Authors: Wansink Brian
Published: June 2021

While dieters may differ, dietary advice too often does not. There were two purposes of this study: 1) Develop a diagnostic means of segmenting dieters, and 2) determine which empirically‐based diet tips are most successful with which segments. The participants consisted of 2500 individuals who had joined the web‐based National Mindless Eating Challenge, expressing an interest in losing weight. After obtaining self‐reported pretest measures of weight and wellness, participants were randomly given three of 30 environmental diet changes (e.g., use smaller bowls, don't serve family style, and so on) that have been empirically suggested to reduce serving sizes or serving frequency.

A majority of dieters lost more weight than those in a face‐to‐face high‐intervention control group. What is remarkable, however, is while some clusters of tips proved to be successful for some segments, other clusters of tips were more successful for other segments. These segmentation differences underscore why many interventions appear to have generally low power: their impact is diffused because it is inappropriate for other segments of dieters.
Black radish and artichoke, for the reduction of Fat Mass
Authors: Figueiredo Marisa Alexandra , Pereira Paula Cardoso
Published: June 2021

Black radish(BR) (Raphanus sativus var Nigra) root has been used in folk medicine since antiquity as a natural drug for the stimulation of bile function. The traditional use of artichoke (Cynara scolymus) fluid extract (AFE) in gastroenterology is mainly based upon its strong antidyspeptic actions which are mediated by its choleretic and hepatoprotective activity. The aim of this study was to investigate the effects of AFE and BR for the reduction of body weigth with the reduction of fat mass. A fraction containing 283,36mg artichoke fluid extract and 522,54mg black radish concentrate are administrated twice a day for seven consecutive days at fifty patients with overweigth and obesity. Participants, on average, lost 0,47kg of their initial weigth and reduced 0,55 kg in fat mass. Although there was an increased of 0,08kg in free fat mass with 0,06kg gain on total body water. This results suggest that black radish and artichoke fluid extract may contribute significantly to weight loss treatment with a reduction on fat mass. However a healthy diet and a regular pratice of exercise do not be excluded in accompaniment to the body weight loss.
Regulation of hepatic lipids and antioxidants by dietary carbohydrate restriction and cholesterol in guinea pigs
Authors: Bruno Richard S , Torres‐Gonzalez Moises , Yeung Steven F , Davis Steven R , Volek Jeff S , Fernandez Maria Luz
Published: June 2021

Carbohydrate restriction (CR) improves parameters of metabolic syndrome and inflammation, but effects on hepatic lipids and antioxidants are less clear. To test the hypothesis that CR favorably affects hepatic oxidative stress and lipids, we assessed hepatic lipids and antioxidants in retired breeder guinea pigs fed a low (L‐) or high (H‐) carbohydrate (CARB) diet with low (0.04%) or high (0.25%) cholesterol (CHOL) for 12 wk (n = 10/dietary treatment). Liver weight was increased by H‐CHOL (p<0.001) and L‐CARB (p<0.05). Guinea pigs fed H‐CHOL had increased (p<0.001) hepatic triglyceride and cholesterol while CARB did not affect hepatic lipids. LCARB increased (p<0.001) hepatic and plasma α‐tocopherol (α‐T) by 1.8 and 4‐times, respectively, whereas H‐CHOL decreased hepatic (p=0.06) and plasma (p<0.05) α‐T by 24% and 54%. Hepatic glutathione ratio (GSH/GSSG) decreased 30% with L‐CARB (p<0.01) and 29% with H‐CHOL (p<0.001), and was due to higher GSSG (p<0.01) and unchanged GSH. GSSG was inversely related (r = ‐0.370; p<0.05) to hepatic α‐T. Hepatic ascorbic acid and uric acid were unaffected by CARB and CHOL. These data suggest that L‐CARB and H‐CHOL result in an oxidative shift of hepatic thiol redox status while L‐CARB improved α‐T status in plasma and liver in aged guinea pigs.
Effects of chromium histidinate supplementation on insulin sensitivity and lipid profile in rat models of insulin resistance and of type 2 diabetes
Authors: Juturu Vijaya , Kazim Sahin , Tuzcu Mehmet , Cikim Gurkan , Komorowski James R
Published: June 2021

The present study evaluated the effects of chromium histidinate (CH) on glucose and lipid metabolism. In vivo models of insulin resistance, and of diabetes, were induced in 8‐week old, male, Sprague Dawley rats by adding a high fat diet (HFD, 40% of calories as fat) for 2 weeks, and by adding a HFD for 2 weeks and then administering a single injection of streptozotocin (STZ, 40 mg/kg i.p.). Rats (∼10/group) were then divided into 6 groups (Control, Control+CH, HFD, HFD+CH, HFD/STZ, HFD/STZ+CH). The control group received a standard diet (12% of calories as fat) and the HFD and HFD/STZ groups continued receiving the HFD. Chromium (Cr) treated rats were fed CH (110 mcg/kg body wt/d). HFD and HFD/STZ treatment reduced serum Cr levels and increased glucose, triglyceride (TG) and total cholesterol (TC) levels (p<0.05). HFD+CH and HFD+STZ+CH groups improved glucose, insulin sensitivity and lipid profile (p<0.05). These results suggest that CH may be beneficial for correcting hyperglycemia and dyslipidemia in insulin resistant and diabetic conditions.
Daily Mg Supplements Above RDA Reduce High Blood Pressure in Hypertensive Human Subjects Not Taking Anti‐hypertensive Medications
Authors: Rosanoff A.
Published: June 2021

BACKGROUND: Studies of magnesium (Mg) therapy for hypertension have led to conflicting results; this study was an attempt to lend clarity to this seeming conflict.

METHODS: Forty‐one human studies testing nutritional Mg supplements with “change in blood pressure” as a measured outcome were collected, sorted as to anti‐hypertensive medication status of subjects, and listed in order of ascending Mg dose.

RESULTS: Daily Mg supplements of 20 mmol or 486 mg/day were necessary to significantly lower high blood pressure in subjects treatment‐naïve or taking anti‐hypertensive medications <6 months. This level is higher than the current adult RDAs for nutritional Mg (320 mg/day for women; 420 mg/day for men). Mg supplements as low as 10 mmol (230 mg) per day significantly lowered blood pressure of hypertensive subjects treated ≥6 months with anti‐hypertensive medications (diuretics, Ca channel blockers, ACE inhibitors, beta‐blockers). Magnesium supplements as high as 25 mmol/day (607 mg) did not significantly lower blood pressure in studies where ≥40% subjects were normotensive at baseline ‐‐ treated or untreated. Mg supplements as high as 40 mmol (972 mg)/day did not lower the blood pressure of subjects speculated to be Mg‐replete by their study's authors, showing the importance of a standard, clinical test for Mg status assessment for full knowledge to come from future studies of nutritional Mg.

CONCLUSIONS: Mg daily dose of 20 mmol (1.1 – 1.5 times higher than RDAs) is necessary to significantly lower high blood pressure in human subjects either treatment‐naïve or on anti‐hypertensive medication <6 months. Subjects’ anti‐hypertensive medication usage ≥6 months (diuretics, beta‐blocker, Ca channel blocker, or ACE inhibitor) reduces by half the daily Mg dose necessary to lower blood pressure in Stage 1, uncomplicated hypertension.
Marginal Biotin Deficiency Successfully Induced by Avidin Supplementation in an Outpatient Design
Authors: Stratton Shawna Lynn , Bogusiewicz Anna , Mock Nell I , Henrich Cindy L , Wells Amanda M , Mock Donald M
Published: June 2021

Previous studies experimentally inducing marginal biotin deficiency have been labor intensive and have utilized an expensive inpatient design. In the current study, we sought to determine the efficacy of inducing marginal biotin deficiency using an outpatient design in which subjects consumed a self‐selected diet. Five healthy adults (3 females) consumed an uncooked egg‐white beverage with all meals and snacks for 28 days. Uncooked egg white contains avidin, which prevents absorption of biotin originating from both the diet and intestinal microbes. A research dietician counseled subjects on avoidance of high biotin foods. Established indicators of biotin status were monitored; urinary excretion of biotin was determined weekly, and lymphocyte activity of the biotin‐dependent enzyme propionyl‐CoA carboxylase (PCC) was determined at two‐week intervals. Urinary biotin decreased steadily and significantly (P< 0.0001 by repeated measures ANOVA); values for 4 of 5 subjects decreased to below the normal range by day 7. PCC activities decreased significantly (P= 0.0052); values for all 5 subjects decreased to below the normal range by day 14. No signs or symptoms of biotin deficiency appeared in any subject. This study provides evidence that successful induction of marginal biotin deficiency is possible in a safe, cost‐effective, outpatient design. NIH DK 36823 and MO1 RR14288
The Vitamin A Nutritional Status of Children Aged 3~12 Years in Hunan Province, China
Authors: Lin Min , Jin Donghui , Li Guangchun , Liu Jiawu , Liu Huilin
Published: June 2021

To understand the prevalence and distribution of vitamin A deficiency in Hunan province among children aged 3~12 years old. 520 children (120 in urban area and 400 in rural area) were examined, selected by stratified multi‐stage cluster sampling in Hunan province. The serum retinol was tested by HPLC and diet data were collected by 3 consecutive 24‐h recalls. The average serum retinol level was 30.5μg/dl, or 36.6μg/dl in urban area and 28.6μg/dl in rural area. The serum retinol levels in all 3~6 age groups were lower than 30.0μg/dl. 6.9% (36/520) of children had serum retinol levels < 0.70 μmol/L (1.7% in urban and 8.5% in rural), 46.0% of children had serum retinol levels between 0.70~1.05 μmol/L (20.0% in urban and 53.8% in rural). There were significant difference in deficiency and marginal deficiency of vitamin A between age groups and genders (X2=7.64, P<0.05; X2=6.892, P<0.05). 74% of children aged 1~13 yrs old consumed vitamin A intakes ≤ 60%RNI, and only 16.5% reached 80%RNI. Vitamin A deficiency is prevalent in children of Hunan province, especially in the rural area. Direct reason caused deficiency of vitamin A is low intake of vitamin A.
A diet high in n‐3 PUFA increases bone strength in broiler chickens
Authors: Davidson Robert T. , Draney Michael L. , Probst Ryan V. , Johnston N. Paul
Published: June 2021

Broiler chickens exhibit a high frequency of leg problems, including fractures, associated with rapid growth. We hypothesized that administering a diet high in vitamin K and flaxseed oil rich in n‐3 PUFA to chickens early in development would improve bone strength. Seventy‐two day‐old male broiler chicks were randomly assigned to one of four groups: 1) control diet (n‐6/n‐3=50:1) containing corn oil and normal vitamin K levels (menadione sodium bisulfite, 22.0 mg/kg diet); 2) high vitamin K diet containing corn oil and high vitamin K (3.3 g/kg diet); 3) flaxseed oil diet (n‐6/n‐3=5:1) containing flaxseed oil and normal levels of vitamin K and; 4) high vitamin K/flaxseed oil diet. Diets were administered six weeks. After the fourth week, six birds from each group were sacrificed each week for the next three weeks. The mechanical strength of the tibiotarsus was determined using a three‐point bending test. BMC was measured using DEXA scanning and bone ashing. Results indicated no significant difference in bone mineral content between treatments. However, flaxseed oil and the combination of flaxseed oil and high vitamin K significantly improved ultimate and yield bending strengths (Dunnett's multiple comparison test p≤0.05). Vitamin K appeared to somewhat reduce the effect of flaxseed oil. The data indicate flaxseed oil may be used to increase bone strength in broiler chickens. Supported by a BYU Mentored Environment Grant.
Short term silicon oral administration increases the bone mineral density in ovariectomized rats
Authors: Kim Mi‐hyun , Bae Yun‐Jung , Chung Yoon‐Sok
Published: June 2021

Silicon may be important for the growth and development of bone and connective tissue. The present study was designed to investigate if soluble silicon(Si) could be used for treatment of postmenopausal osteoporosis. Soluble silicon (Si 20mg/kg body weight/d) was orally administrated to 17 weeks‐old ovariectomized rat for 4 weeks. Silicon did not alter weight gain in OVX rats. Silicon supplementation significantly increased the bone mineral density(BMD) of femur(p<0.05, vs OVX control group) and tibia in OVX rats(p<0.05, vs OVX control group). But, serum CTx(C‐telopeptide cross‐links of type I collagen), bone resorption biomarker, was significantly increased in silicon supplemented group. Serum ALP(Alkaline phosphatase) and osteocalcin, bone formation biomarker, were not significantly changed by treatment with silicon in OVX rats. According to these results, the silicon supplementation resulted in increasing the bone mineral density in OVX‐induced osteoporosis, and the effects might be mediated by other mechanism exterior to bone turnover.
Grifola frondosa inhibits TNF‐α‐induced intestinal inflammation via suppression of MCP‐1 and IL‐8 expressions in HT‐29 human colon epithelial cells
Authors: Park Su‐Young , Lee Jong Suk , Thapa Dinesh , Park Byung Chul , Kim Ah Ra , Choi Mi Kyoung , Kim Jung‐Ae
Published: June 2021

Grifola frondosa (GF), a king‐size edible mushroom, has been used as a health food for centuries in China and Japan. In the present study, we investigated an anti‐inflammatory action of GF in inflammatory bowel disease (IBD) by using HT‐29 human colon epithelial cells stimulated with TNF‐α to induce intestinal inflammatory condition in vitro. Treatment with GF significantly prevented the attachment of leukocytes to HT29 colon epithelial cells, one of the pathologic hallmarks in colon inflammation. The effect of GF was comparable to that of 5‐aminosalicylic acid, a commonly used drug in the treatment of IBD. In addition, pretreatment with GF inhibited TNF‐α‐induced MCP‐1 mRNA expression and protein secretion. Furthermore, GF reduced the TNF‐α‐induced interleukin‐8 mRNA expression. These results demonstrate that GF modulates intestinal inflammation via suppressing the expression of inflammatory cytokines, and suggest that GF may be a valuable medicinal food for prevention and/or attenuation of IBD.
MDI 301, A non‐irritating retinoid, improves abrasion wound healing in both aged and diabetic skin
Authors: Warner Roscoe L. , Bhagavathula Narasimharao , Hanosh Andrew , McClintock Shannon D. , Naik Madhav K. , Johnson Kent J. , Varani James
Published: June 2021

MDI 301 is a picolinic acid‐substituted ester of retinoic acid (RA) and has been shown to increase epidermal thickness and pro‐collagen synthesis in organ‐cultured human skin. Unlike RA, MDI 301 does not induce expression of pro‐inflammatory cytokines or induce expression of leukocyte adhesion molecules. In the present study topical MDI 301 improves the structure and function of skin in two models of skin damage in rodents and improves dermal wound healing in these models. MDI 301 was applied daily to the skin of diabetic rats (eight weeks post treatment with streptozotocin) and to the skin of aged (14–16 month old) rats. In both models, subsequently‐induced abrasion wounds healed more rapidly in the retinoid‐treated animals than in vehicle‐treated controls. Immediately after complete wound closure, tissue from the wound site (as well as from a control site) was put into organ culture and maintained for three days. At the end of the incubation period, culture fluids were assessed for soluble type I collagen. In both models, the level of type I collagen was increased by MDI 301, while the levels of skin irritation was virtually nonexistent. These findings demonstrate that a MDI‐301 has the beneficial effects of increasing skin healing without the traditional ill effects of skin irritation and offers a potential substitute for the parent Retinoic Acid. Work supported by NIH RO1 HL07097
Sulfiredoxin is Regulated by NRF2 During Oxidative Stress in Lungs
Authors: Ling Guoyu , Sussan Thomas E , Thimmulappa Rajesh K , Biswal Shyam
Published: June 2021

Mammalian sulfiredoxin (SRX) plays a role in the regulation of oxidative stress. However, the transcriptional regulation of SRX in response to oxidative stress is not clear. Here we demonstrated that NRF2 transcription factor regulates SRX expression following oxidative stress in lungs. In response to cigarette smoke exposure, lungs from wild‐type mice showed significant increase in the mRNA and protein expression of SRX, while no induction was observed in lungs from NRF2‐/‐ mice. Similarly, diquat‐induced oxidative stress resulted in increased expression of SRX in wild‐type mouse embryonic fibroblasts (MEF), but not in the NRF2‐/‐ MEF cells. Silencing SRX by siRNA induced greater generation of ROS in Raw 264.7 macrophages. Promoter analysis revealed two antioxidant response elements (ARE) in the mouse SRX promoter. Transient transfection studies, using reporter constructs containing the SRX promoter, in NRF2‐/‐, wild‐type, and KEAP1‐/‐ MEF cells demonstrated low, modest and high reporter activity, respectively, suggesting NRF2‐dependent transcriptional regulation. Chromatin immunoprecipitation assays revealed binding of NRF2 to the ARE element upon hydrogen peroxide treatment. Taken together, our data demonstrates that NRF2 regulates basal and inducible expression of SRX in response to oxidative stress, which may have an important role in cell signaling and protection.
Mechanisms of bradykinin‐induced expression of vascular cell adhesion molecule‐1 in human rheumatoid arthritis
Authors: Wu Pei‐Rung , Yang Chuen‐Mao
Published: June 2021

Increased expression of adhesion molecules by inflammatory mediators such as bradykinin (BK) plays a key role in inflammatory diseases during rheumatoid arthritis (RA). However, the mechanisms by which BK induced expression of vascular cell adhesion molecule‐1 (VCAM‐1) remained unknown in RA. Here, we reported that in human synovial fibroblasts, BK‐induced expression of VCAM‐1 protein, mRNA, and promoter gene activity was attenuated by the inhibitors of MEK1/2 (U0126), p38 MAPK (SB202190), JNK (SP600125), and NF‐κB (Bay11‐7082). Transfection with dominant‐negative plasmids of MEK, ERK1/2, p38, JNK, NIK, IKKα or IKKβ also abolished the VCAM‐1 expression. Accordingly, BK‐stimulated phosphorylation of p42/p44 MAPK, p38 MAPK, and JNK was also attenuated by pretreatment with U0126, SB202190, and SP600125, respectively. Moreover, BK‐induced NF‐κB activation was attenuated by pretreatment with U0126, SB202190, SP600125, or Bay11‐7082, determined by immunofluorescence staining. Furthermore, BK‐stimulated monocytes adhering on synovial fibroblasts via VCAM‐1 expression was decreased by pretreatment with U0126, SB202190, and SP600125. These results indicated that activation of ERK1/2, p38, JNK, and NF‐κB were involved in VCAM‐1 expression.
Immunohistochemical Localization and Multi‐gene Expression Profiling of O2 Sensor Components in Airway Chemoreceptors
Authors: Pan Jie , Yeger Herman , Cutz Ernest
Published: June 2021

Pulmonary neuroepithelial bodies (NEB) are airway chemoreceptors that express a membrane bound O2 sensor molecular complex (NADPH oxidase coupled to an O2 sensitive K+ channel). We used confocal immunofluorescence microscopy with specific antibodies against NADPH components and O2 sensitive K+ channels (K+O2) to label NEB in rat /rabbit lung and NEB related H146 tumor cell line. For gene expression profiling of NEB cells microdissected from human lung, and H146 cells, we used a custom MultiGene‐12TM RT‐PCR array that included NADPH oxidase complex and homologues /accessory proteins (NOX1‐4, phox‐p22, p40, p47, p67, Rac1, NOXO1 and NOXA1) and K+O2 channels (TASK1‐3, Kv ‐1.2,1.5, 2.1, 3.1, 3.3, 3.4, 4.2, 4.3). In rat lung, NOX2, NOX4, p22phox, TASK1 and Kv3.3 (and Kv3.4 in rabbit) localized mainly to the apical plasma membrane of NEB cells, and membrane or sub‐membrane regions in H146 cells. NOX2 co‐localized with p22phox, p40phox, p47phox and Kv3.3, while NOX4 co‐localized with TASK1. Gene expression profiling of NEB/H146 cells confirmed the presence of all NOX proteins except NOX3 and NOXO1, as well as all common K+O2 channels, except Kv1.5 and Kv4.3. Adjacent airway epithelial cells expressed only TASK2, Kv2.1, Kv3.4 and Kv4.2. Present findings support the plasma membrane model of O2 sensing by airway chemoreceptors that appears to be cell and organ specific. (Supported by grants from CIHR)
Adenohypophysitis in rat pituitary allografts.
Authors: Rotondo Fabio , Quintanar‐Stephano Andres , Lombardero Matilde , Asa Sylvia L , Kovacs Kalman
Published: June 2021

The histologic changes in pituitary allografts transplanted under the kidney capsule from Wistar to Lewis rats were investigated. Normal pituitaries, pituitary autografts and intrasellar pituitaries of rats bearing allografts were also examined. Rats were sacrificed at various times after transplantation. Sections stained with H & E were investigated by light microscopy. Two days after transplantation, the central part of the allografts showed ischemic necrosis. A week later, massive mononuclear cell infiltration composed mainly of lymphocytes was seen. Several weeks after transplantation, the mononuclear cell infiltration decreased, the surviving adenohypophysial cells increased in number and the necrotic area was replaced by connective tissue. In the autografts the central ischemic necrosis was similar to that of the allografts. The mononuclear cell infiltration was, however, mild and the adenohypophysial cells seemed to increase in number earlier than those of the allografts. No histologic changes were noted in control adenohypophyses and in intrasellar pituitaries of rats bearing allografts. It appears that pituitary allografts are “foreign” and evoke an autoimmune response. Pituitary allografts represent useful experimental models to investigate the development and morphology of adenohypophysitis, a rare disease occurring mainly in young women often associated with pregnancy.
Pathway Analysis of Global Gene Expression Changes in rat liver following silencing of c‐met by RNA Interference.
Authors: Paranjpe Shirish , Tseng George , Chattopadhyay Ansuman , Luo Jianhua , Michalopoulos George K
Published: June 2021

We have reported Cell cycle effects resulting from shRNA mediated inhibition of hepatocyte growth factor and its receptor c‐Met in regenerating rat livers following a partial hepatectomy (Hepatology, 45: 1471–1477). Microarray analysis of shMet and scrambled treated rat livers using Affymetrix U230.2 A chip containing 10,000 expressed sequences indicated abnormal expression of genes associated with cell cycle and apoptosis. Cluster analysis of microarray data was then carried out. Normalization and preprocessing of data from 8 chips was by dChip software. Expression intensities were log transformed and genes with expression level less than 7 or SD smaller than 0.5 were filtered out. Hierarchical clustering of the remaining 4747 genes according to similarity in pattern of gene expression produced 8 clusters. Of the 4747 genes, 805 genes whose expression value was >2 or <–2 compared to the corresponding control group were then analyzed using Ingenuity software. Networks were generated for each time point post PHx for shMet treatment and control group.

It was observed that the genes associated with DNA repair, fatty acid metabolism, p38 MAPK pathway, and bile acid biosynthetic pathway were differentially regulated in shMet silenced rats when compared to scrambled treated controls.

Suppression of c‐met thus caused profound alterations in multiple signalling pathways.
Beta‐catenin regulates proliferation and differentiation of oval cells in mouse liver
Authors: Thompson Michael David , Cieply Ben , Singh Sucha , Monga Satdarshan PS
Published: June 2021

Atypical ductular proliferation (ADP) occurs in the liver following DDC‐induced liver injury. We noted blunted ADP in mice liver lacking β‐catenin. Also based on the similarities between the processes of ADP in mice and oval cell response in rats, and the role of Wnt/β‐catenin signaling in both events, we sought to examine whether presence of active‐β‐catenin would provide a growth or differentiation advantage to atypical ductular cells in mice. Liver‐specific β‐catenin over expressing mice (TG) with a stabilizing mutation at ser45, were fed the DDC diet. No significant differences in appearance and numbers of A6‐positive cells were observed. Intriguingly, increased numbers of A6‐positive cells with hepatocyte‐like morphology were observed in TG mice. Contrary to short‐term exposure (<4wk) in β‐catenin knockout mice, long‐term exposure to DDC (5 months) led to a significant increase in A6‐positive cells with bridging fibrosis indicating their continued expansion and lack of differentiation. In sum, these findings indicate diverse roles Wnt/β‐catenin signaling during oval cell response in mice.
Interactions of β‐catenin with Met & E‐cadherin during Oval Cell Activation.
Authors: Apte Udayan , Stolz Donna B. , Monga Satdarshan P. S.
Published: June 2021

We have previously demonstrated that canonical Wnt/β‐catenin pathway plays a critical role in activation and proliferation of hepatic progenitor cells or oval cells in rats and mice. Here, we investigate if non‐canonical modes of β‐catenin activation play a role in oval cell emergence, expansion or differentiation in rats in the 2‐AAF+PHX models. Confocal microscopy identified Met‐β‐catenin (M‐B) complex apically and E‐cadherin‐β‐catenin (E‐B) on all surfaces in normal hepatocytes. Both these complexes were found on all surfaces in biliary epithelia. In rats treated with 2AAF+PHX, both complexes were observed in oval cells, in a biliary pattern, at 5 days after PHX. However, at 10 days post‐PHX, a clear decrease in both M‐B and E‐B complexes was observed that coincided with nuclear translocation of β‐catenin and active proliferation of oval cells. At 20 days post‐PHX, robust reappearance of both complexes was evident in differentiating oval cells that coincided with decreased β‐catenin activation. Thus, β‐Catenin complex with Met and E‐cadherin undergoes notable modulation during oval cell expansion and during their differentiation into hepatocytes. To conclude, while canonical β‐catenin activation plays a role in oval cell emergence, this is supplemented by noncanonical means during oval cell expansion and differentiation.
Regucalcin/SMP‐30 is regulated by β‐catenin in the liver
Authors: Nejak‐Bowen Kari , Zeng Gang , Monga Satdarshan P
Published: June 2021

Our laboratory has identified regucalcin as a novel target of β‐catenin in the liver. We show that regucalcin mRNA levels were downregulated in β‐catenin conditional knockout mice (KO), as compared to their wild‐type (WT) controls. Immunohistochemistry (IHC) and Western blotting (WB) confirms that regucalcin protein expression is also decreased in these mice. Analysis of HepG2 and Hep3B hepatoma cell lines demonstrates that HepG2 (active β‐catenin) have higher levels of regucalcin as compared to Hep3B (normal β‐catenin). Further, we find a correlation between the presence of β‐catenin in the nucleus or cytoplasm of human hepatocellular carcinomas (HCC) and an increase in regucalcin by IHC and WB. An analysis of the promoter region of the mouse regucalcin gene reveals at least 11 putative Tcf‐binding sites. Use of multiple deletion mutants identifies the specific sites, which might be regulating regucalcin expression by β–catenin. Finally, we examine the regucalcin expression in methionine‐choline‐deficient (MCD) diet‐induced steatohepatitis. IHC identifies an increase in regucalcin levels in MCD‐fed mice, as compared to control diet‐fed mice. In conclusion, regucalcin is a novel target of β‐catenin levels in liver and may have a biological function in liver diseases such as steatohepatitis and HCC.
Ramelteon Phase Shifts Circadian Rhythms Of Activity And Promotes Sleep In The Rhesus Monkey
Authors: Dubocovich Margarita L , Lambert Aaron
Published: June 2021

This study assessed the effect of the MT1/MT2 melatonin receptor agonist ramelteon on sleep, measured as changes in immobility time and on phase shift of the onset of circadian rhythm of motor activity determined by actigraphy in Rhesus monkeys. Monkeys free run, under constant dim illumination (5–15 lux), with a circadian period (tau) of 23.8 ± 0.03 h. Ramelteon (300 ug/kg, oral) (from RozeremTM tablets) given 9‐10 h after onset of activity for 3 consecutive days induced a robust phase advance of the onset of circadian rhythm of activity [1.32 ± 0.19 h (n=6) vs. 0.02 ± 0.05h (n=6) for placebo, p < 0.05] with no change in tau. Ramelteon induced a permanent phase advance since the onset of circadian rhythm of activity remained entrained for at least 9 days. In monkeys maintained in a 14h dim light (5–15 lux) /10h dark cycle ramelteon (300 ug/kg, oral) given 9 h after onset of activity significantly increase % sleep (mean±SEM, n=11] with a maximum effect within 1 h after administration (84.4 ± 6.7 vs. 49.1 ± 4.1 % sleep/h/3days, p < 0.001]. For a ramelteon dose of 30 ug/kg the sleep promoting effect was statistically significant until 4 h after administration. The effect of ramelteon on sleep was not observed the day after the last dose suggesting that ramelteon promotes sleep and phase shift circadian rhythms of activity by activation of different receptors and/or cellular mechanisms. Supported by MH 63466 (MLD).
Realization and validation of cellular sensors to detect acetylcholine in vivo
Authors: Nguyen Quoc‐Thang , Schroeder Lee Frederick , Taylor Palmer W , Mank Marco , Griesbeck Oliver , Kleinfeld David
Published: June 2021

In vivo detection of neurotransmitters with high temporal and spatial resolution is critical for understanding brain physiology. We adapted the use of the fluorescent cell‐based assay to create implantable Sentinel Cells to detect acetylcholine (ACh). Functional signals from these cells were imaged in vivo with two‐photon laser scanning microscopy (TPLSM).

Sentinel cells were created by stably expressing the IP3/Ca2+‐coupled M1 muscarinic receptor and the genetically encoded FRET‐based Ca2+ sensor TN‐XXL into HEK293 cells. In vitro, bath application of ACh elicited a FRET signal in Sentinel Cells which increased monotonically from a treshold of ~ 30 nM (~ 30 % change in 7 s) to saturation at ~ 1 μM ACh (> 90 % change in 5 s).

Sentinel cells were implanted in rat neocortex down to 200 μm from the surface and imaged by TPLSM. Acutely implanted cells had normal morphology and responded for at least 6 hours after implantation. Endogenous release of ACh was generated by electrical stimulation of the nucleus basalis magnocellularis which triggered signal changes (≥ 50%) in Sentinel cells. These responses were enhanced with the systemic injection of either tacrine or physostigmine. Following chronic intracerebral implantation, imaging of Sentinel cells could take place for up to 5 days.

Our technique could potentially detect neurochemicals other than ACh by using different receptors or reporters.
Nicotine‐Ethanol Interaction: Regulation by two distinct nicotinic acetylcholine receptor (nAChR) subtypes in mouse cerebellum
Authors: Taslim Najla , Dar M. Saeed
Published: June 2021

We previously reported cross tolerance between nicotine and ethanol and attenuation of ethanol ataxia by intracerebellar (ICB) nicotine. Using rotorod and direct ICB infusion in cannulated mice, we studied the role of nAChR subtypes á4â2 and á7. RJR‐2403 [32.25, 62.5, 125 ng] a á4â2 agonist, markedly attenuated ethanol (2g/kg; i.p) ataxia dose dependently. Dihydro‐â‐erythroidine, DHâE, á4â2 antagonist, (500ng) significantly attenuated RJR's and nicotine‐induced attenuation of ethanol ataxia reinforcing the role of á4â2 subtype in nicotine‐ethanol functional interaction. Also, ICB infusion of PNU 282987 (250ng–2.5ug), a novel á7 subtype selective agonist reduced ethanol ataxia thus indicating the probable role of á7 nAChR subtype in this context. Moreover, pretreatment with methyllylcaconitine (MLA; 3–12ng; ICB), a á7‐subtype selective antagonist, effectively prevented PNU and nicotine attenuation of ethanol ataxia confirming the contribution of á7 subtype in ethanol's ataxia. Both DHâE and MLA per se had no effect on ethanol's ataxia precluding any tonic role of nAChR subtypes. Taken together, our results support a comparable role of nAChR á4â2 and á7 subtype in nicotine‐induced attenuation of ethanol ataxia. Future treatment strategies for ethanol ataxia might require manipulation of these nAChR subtypes to lessen alcohol related motor impairment.
Signaling Mechanisms Responsible for Muscarinic Potentiation of GABAergic Inputs to Spinal Dorsal Horn Neurons
Authors: Zhang Hongmei , Richardson Timothy E , Chen Shaorui , Pan Huilin
Published: June 2021

Activation of muscarinic acetylcholine receptors (mAChRs) potentiates spinal GABAergic tone, which is one of the potential mechanisms of muscarinic analgesia in the spinal cord. Although M2, M3, and M4 subtypes are all involved in the potentiation of spinal GABA release, the signaling mechanisms of mAChRs involved in this action are not clear. By whole‐cell recording of GABAergic spontaneous inhibitory postsynaptic current (sIPSCs) in lamina II neurons in the rat spinal cord slice, we found that the stimulating effect of the specific mAChRs agonist oxotremorine‐M on GABAergic sIPSCs was abolished in the Ca2+‐free extracellular solution. Blocking of voltage‐gated Ca2+ channels with Cd2+significantly reduced the effect of oxotremorine‐M on sIPSCs. However, neither the KCNQ channel blocker XE991 nor the adenylate cyclase inhibitor MDL‐12330A affected the effect of oxotremorine‐M on sIPSCs. Blocking of the non‐selective cation channels with SKF96365 or 2‐APB significantly inhibited the effect of oxotremorine‐M on sIPSCs. Treatment with the phosphoinositide‐3‐kinase inhibitor wortmanin or LY‐294002 abolished the stimulating effect of oxotremorine‐M on sIPSCs in 68% neurons tested. In the spinal cord slices from pertussis toxin treated rats (to eliminated M2 and M4 subtypes‐mediated effect), wortmanin also blocked the effect of oxotremorine‐M on sIPSCs in 65% neurons. Therefore, this study provides new information that mAChRs stimulate GABAergic neurons by Ca2+ influx through voltage‐gated Ca2+ channels and phosphoinositide‐3‐kinase ‐nonselective cation channel pathway in the spinal dorsal horn.