bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of angiotensin II increases hypothalamic thyroid hormone and stimulates orexigenic NPY/AgRP neurons (LB437)
Authors: Wang CHih‐Hong , Smithies Oliver , Takahashi Nobuyuki
Published: June 2021

We have demonstrated that mice lacking renin (Ren1c ‐/‐) are lean and resistant to diet‐induced obesity, which is associated with increased energy expenditure and uncoupling proteins (UCPs) expression. Surprisingly, plasma catecholamine levels of Ren1c‐/‐ mice were similar to wild type (WT) mice, and their plasma total T3 and T4 were lower than those of WT, although they showed no obvious signs of hypothyroidism. We here investigate whether mice lacking renin display true hypothyroidism, and if so, the specific mechanism by which they develop it. The Ren1c‐/‐ mice have approximately 50% levels of free T3, freeT4 and TSH in the plasma, as well as reduced TRH expression in the hypothalamus compared to WT. Administration of TRH or angiotensin II (Ang II) both restored plasma TSH and thyroid hormones in the Ren1c‐/‐ mice, while these hormones were not affected by human renin, which stimulates mouse MAP kinases but does not cleave mouse angiotensinogen to Ang I. Moreover, the Ang II receptor blocker losartan decreased TRH, TSH, and thyroid hormones of WT mice to the levels similar to those in mice lacking renin. Furthermore, mice lacking renin and WT mice treated with losartan have significantly increased hypothalamic expression of type 2 deiodinase (Dio2), which converts T4 to the more potent T3, and have increased thyroid hormones in the hypothalamus, which likely inhibits TRH through feedback. Both mice lacking renin and WT mice treated with losartan have 3‐fold higher uncoupling protein‐2 (UCP2), orexigenic neuropeptide Y (NPY) and agouti‐related protein (AgRP), as well as 5‐fold lower anorexigenic proopiomelanocortin (POMC) gene expression in the hypothalamus relative to untreated WT mice. In contrast, Ren1c‐/‐ mice treated with human renin had similar expression levels of these genes. These results suggest that the inhibition of Ang II not of Ang‐independent effects of renin causes hypothyroidism and enhances food intake, which likely decrease energy expenditure. We hypothesize that changes in gene expression of the hypothalamus do not contribute to increased energy expenditure of the Ren1c‐/‐ mice, and that hypothalamic hyperthyroidism feedback inhibits TRH, leading to systemic hypothyroidism. Caution should be paid to hypothyroidism when using high dose of inhibitors of the renin angiotensin system.

Grant Funding Source: National Science Council: 102‐2320‐B‐009 ‐002 ‐MY3
Effects of enzyme co‐consumption with whey protein in adult males (LB442)
Authors: Maher Margaret , Kobs Lisa
Published: June 2021

Whey protein supplements are used by athletes to promote muscle protein synthesis and reduce protein degradation and may also benefit persons with muscle wasting conditions. Unfortunately, digestion of whey protein may liberate short peptides that can cause immune‐mediated gastrointestinal (GI) distress. Co‐consumption of whey with enzymes may result in smaller and less immunogenic peptides. In this study, sixteen males (ages 18‐27 years) consumed whey only (WO) and whey plus enzymes (WE) in a randomized cross‐over design. Subjects underwent 6‐day dietary preconditioning with standard meals supplied to them (approximately 2300 kcal, 44% carbohydrate; 21% protein; and 35% fat). Following a 12‐hour fast prior to each treatment trial, serum was collected before (time 0) and 1, 2, 3, and 4 hours after consumption of WO or WE. Serum was analyzed for amino acids, insulin, blood urea nitrogen (BUN), and C‐reactive protein (CRP). Subjects also completed a brief survey assessing GI symptoms at approximately 4 hours. Two‐way repeated measures statistics, with within factors of time and treatment (WO and WE) and between factor of treatment order were performed. Areas under curves were also analyzed with paired samples T‐tests. There were significant time and treatment effects with several amino acids and time by treatment by order effects for a few. An expected increase (p<0.016) in insulin and BUN following supplement consumption was observed but there was no difference in the insulin or BUN response between WO and WE. CRP levels were highly variable across subjects. CRP decreased (p<0.048) over time with WE, while there was no significant change in CRP over time with WO. Co‐consumption of whey with enzymes results in altered amino acid appearance and/or disposal in serum and may reduce acute stress as measured through CRP. No significant impact of either WO or WE on GI symptoms was observed in this sample.

Grant Funding Source: Supported by a grant from Deerland Enzymes Inc.
Effect of a very low carbohydrate diet followed by incremental increases in carbohydrate on respiratory exchange ratio (LB444)
Authors: Kunces Laura , Volk Brittanie , Freidenreich Daniel , Saenz Catherine , Fernandez Maria Luz , Maresh Carl , Kraemer William , Phinney Stephen , Volek Jeff
Published: June 2021

Low carbohydrate diets are associated with increases in fat oxidation, but the extent to which substrate utilization is altered as individuals reintroduce carbohydrate into the diet has not been studied under controlled conditions. We therefore examined postabsorptive respiratory exchange ratio (RER) across a broad range of dietary carbohydrate and fat levels in adults with metabolic syndrome. After an initial 3‐wk run‐in low‐carbohydrate diet, 16 adults (age 55 ± 10 yr, BMI 37.9 ± 6.3 kg/m2) were fed six sequential moderately hypocaloric 3‐wk diets that progressively increased carbohydrae (47, 82, 131, 179, 250, 344 g/day) with parallel decreases in fat. Resting RER was lowest after the very low carbohydrate diet and increased in a linear manner as carbohydrate increased (0.75 ± 0.04, 0.77 ± 0.03, 0.79 ± 0.04, 0.80 ± 0.04, 0.82 ± 0.03, 0.84 ± 0.05, respectively). These findings indicate that very low carbohydrate diets accelerate fat oxidation and that progressive increases in carbohydrate track linearly with decreased fat oxidation.
Acarbose effects on energy balance, body composition, and tissues: A caloric restriction mimetic? (LB448)
Authors: Brewer Rachel , Miyasaki Nathan , Smith Daniel
Published: June 2021

A recent study has shown that administration of acarbose (ACA), an α‐glucosidase and α‐amylase inhibitor approved for use in type 2 diabetes, led to increased lifespan in mice. The mechanism for this lifespan increase is unclear; however, by reducing carbohydrate metabolism, we hypothesize that ACA acts in a manner similar to caloric restriction. To test this hypothesis, a six‐month dose‐response study was performed in both male and female C57BL/6J mice treated with ACA. Groups of mice (n=9‐10) received diet with ACA at doses of 0.05% (low), 0.1% (medium), or 0.15% (high), as well as a no‐dose control. High‐dose ACA administration in male mice led to 10% lower body weight (p<0.01) and >50% smaller fat depot size (p<0.001) versus control, with similar decreases in both visceral and subcutaneous depots and no significant difference in lean mass. Total energy expenditure and activity were not different between groups, while respiratory exchange ratio tended to be lower with high‐dose ACA administration (p=0.06). In contrast to body weight, a 50% increase in food intake was observed with high‐dose ACA (p<0.001), along with a >300% increase in fecal output wet weight (p<0.001). Both small and large intestine increased >20% (p<0.001), while total filled caecum wet weight increased by >200% (p<0.001). Female mice responded similarly to males (excluding body weight differences), with a dose dependent response for multiple measures in both sexes. Similar to caloric restriction, ACA administration led to smaller fat depot size and lower body weight, though gastrointestinal changes suggest alterations to carbohydrate metabolism independent of traditional caloric restriction. Comparisons with glycemic matched diets and total caloric restriction are being further investigated.

Grant Funding Source: Supported by The Ellison Medical Foundation New Scholar in Aging Award (D. Smith), UAB Neuroscience Behavioral Assessment Core (P30 NS47466), and UAB Small Animal Phenotyping Core (P30 DK056336)
Transitions in dietary patterns among adults in South Korea from 1998‐2010: baseline examination of the Severance cohort study (LB454)
Authors: LIM HYUNJUNG , Kim Sang Yeun , Wang Youfa , Lee Sun Ju , Oh Kyungwon , Sohn Chun Young , Moon Young Myoung , Jee Sun Ha
Published: June 2021

Transitions in nutrition patterns tend to emerge through industrialization and economic development. We hypothesized that dietary patterns among adults aged蠅20 years in South Korea have changed significantly from 1998 to 2010. We used a repeated cross‐sectional data from baseline examination of the Severance cohort study wherein 134,366 adults were followed annually. We determined changes in consumption, adjusted for age and sex, and tested the trends across the study period. Factor analysis and cluster analysis were used to derive dietary patterns. The consumption of traditional Korean foods including cereals, vegetables (377g to 256g), and Kimchi (127g to 85g) decreased, while fruit (170g to 251g), dairy, egg, and fried food intakes increased (P<0.05). Total daily energy intake declined steadily from 2003 to 1688 kcal. Carbohydrate intake declined by 17.5% over the study period; however, fat‐derived energy intake increased from 18.6% to 20.1% (P<0.05). Our factor and cluster analyses identified 3 dietary patterns that we termed ‘Korean’ diet (rice, vegetables, and Kimchi), ‘Western’ diet (soda, eggs, and oil) and ‘Well‐being’ diet (low sugar and high fruit and dairy product intakes). Compared to 1998, approximately 40% of participants still followed a Korean diet in 2010. Interestingly, the popularity of the Western diet fell by around a fifth, whereas the Well‐being diet increased two‐fold over the study period. This demonstrates a unique transition in food and nutrient intakes in Korea since 1998. We found secular trends in dietary patterns that included a preservation of the traditional Korean diet and the emergence of a Well‐being diet.
Diet and nutritional status of middle aged and elderly residents of Urumqi community (LB455)
Authors: Han Jia , LI Cai Hong , Yuan Qian Qian , Xu Shu , Chen Roujin , Abuduli Wuhuliayi , Xiaerpi Wueryetiguli
Published: June 2021

Objective A community‐based study was undertaken in two communities to assess the diet and nutritional status of middle‐aged and elderly residents of Urumqi, Xinjiang. Methods The 24‐hour food intake method combined with food frequency questionnaire were used to evaluate the food intake of 579 middle‐aged and elderly residents aged 40‐76 years.Results The major type of food for the residents surveyed was plant‐based foods. Intakes of cereals, vegetables, fruits, poultry and eggs either reached or nearly reached with narrow margins the standards set by the Chinese Food Guide Pagoda (2007). However, intakes of fat and salt were 39.0g /d and 12.1g/d respectively, exceeding the recommendations of Chinese Food Guide Pagoda. Energy intakes from protein and fat were 15.9% and 38.2%, respectively, higher than recommendations. Carbohydrates‐derived energy was however below the recommended ratio (45.9%). Average intake level of calcium was 614.7mg/d, well below the RNI(Recommended Nutrient Intake). Although iron intake reached the RNI , 77.1% of iron was derived from plant foods. There was no statistical difference between genders. Conclusion The dietary status of middle‐aged and elderly residents of Urumqi community based on intakes of macronutrients and select micronutrients was not satisfactory according to the Chinese Food Guide Pagoda. There is a need for improving the quality of the diet and nutrition education of these residents, particularly from the perspective of chronic disease prevention.

[Key words] middle‐aged and elderly residents; dietary intake; nutrients

Grant Funding Source: Supported by NSFC
Determinants of anemia among school‐aged children in Colombia and the United States (LB456)
Authors: Syed Sana , Addo O. Yaw , Ziegler Thomas , Suchdev Parminder
Published: June 2021

Anemia globally affects a quarter of all school‐aged children (SAC) and is frequently ascribed to iron deficiency (ID). Other nutritional, demographic and socio‐economic factors are often unmeasured. We obtained data from two national cross‐sectional surveys – 2010 Encuesta Nacional de Nutrición Situación Colombia and the 2003 to 2006 US National Health and Nutrition Examination Survey. We analyzed data for 10570 Colombian and 3605 US children aged 5 to 15 years. Associations were studied using survey regression methods with adjustment for inflammation, potential confounders and complex survey design effects. Colombian data showed 51% males, 85% non‐minority status, 13% stunted, 4% wasted, 4% overweight, 0.5% obese. Prevalence of anemia was 1.6%; ID was 9.2%. US data showed 52% males, 59% non‐Hispanic White, 2% stunted, 2% wasted, 19% overweight, 5% obese. Prevalence of anemia was 0.8%; ID 12%. In Colombia, anemia was associated with serum ferritin levels (OR 0.98, 95%CI=0.96–1.00, p=0.056) and socio‐demographic factors (age p<0.001, gender p=0.04, ethnicity p=0.02). In the US, anemia was associated with soluble transferrin receptor levels (OR 2.3, 95%CI=1.5–3.7, p<0.001), serum retinol levels (OR 0.06, 95%CI 0.01–0.28, p<0.001) and ethnicity (p<0.001). Growth was not associated with anemia in either country. Prevalence of anemia among SAC in both countries was extremely low. In general, ID was associated with anemia with inconsistent associations with socioeconomic factors between countries which need to be explored further. Additional countries (Malawi and Pakistan) are planned for future analyses.

Grant Funding Source: Supported by Bill & Melinda Gates Foundation, CDC, GAIN, NICHD, NIH ACTSI.
A Comparison of Nutritional Status of Children Under the Age of Five Years, in 1999 and 2010 in Matobo District, Zimbabwe. Results From a district survey and the National Nutrition Survey. (LB458)
Authors: Katuli Sozina , Sun Xuyang , Husain Zuhair , Cordero‐MacIntyre Zaida , Beeson Lawrence
Published: June 2021

This study compares nutritional status of children under‐five years in two time periods using results of January 1999 district survey and January 2010 National Nutrition survey.

Objectives. To explore the changes of nutritional status of children 6‐59 months in 1999 and 2010 and identify underlying reasons for the changes.

Methods. 614 children were evaluated in 1999 and 563 were evaluated in 2010. We used 30 x10 cluster in 1999 and 30 x 20 cluster design in 2010. Thirty clusters were selected from all villages in Matobo district and ten household surveyed in 1999 and 20 households were surveyed in 2010. Data were collected using a semi‐structured questionnaire and children were weighed and measured for nutrition assessment using WHO standards for stunting, wasting and underweight.

Results: We observed a significant improvement in nutritional status during the two time periods. Severe (< ‐3 z‐scores stunting was reduced from 10% in 1999 to 5.5% in 2010; below cut off stunting decreased by 5.3%. Severe wasting remained unchanged while below cut off (< ‐2 z‐scores) wasting decreased by 2.9%. Severe underweight decreased by 3.6%) underweight below cut off (< ‐2 z‐scores) declined by 4.3%.

Conclusion: Acute malnutrition improved over the 10 year period. However, chronic malnutrition slightly increased possibly due to food insecurity.

Key words: nutritional status, children under five, cluster survey, WHO standards

Grant Funding Source: Zimbabwe Ministry of Health and Social Walfare
Contextual and cultural influences on parental feeding practices among Hispanics (LB465)
Authors: Mena Noereem , Gorman Kathleen , Greene Geoffrey , Tovar Alison
Published: June 2021

Contextual and cultural influences on parental feeding practices among Hispanics Hispanic preschoolers in the United States are at greater risk for obesity compared to non‐Hispanic whites. Obese children are more likely to be obese adolescents and adults thus, early prevention is critical. Growing evidence has addressed the role of caregivers’ indulgent or restrictive feeding practices in children’s eating behaviors and negative weight outcomes. However, the mechanisms driving these associations are largely unknown; including what influences these feeding practices among Hispanics. Therefore the goal of this qualitative study was to explore contextual and socio‐cultural influences on parental feeding. Four focus groups (n=37) were held with Hispanic parents of preschool‐aged children at two day care centers in Rhode Island. Audio recordings were transcribed and translated, and themes extracted. Parents commented that their childhood experiences of not being allowed to waste food influenced how they currently feed their children. They reported making conscious decisions not to use the same feeding strategies as their own parents and on the contrary, reported using different practices related to encouragement of healthy foods and promotion of self‐regulation. Parents reported wanting to feed their children healthy foods but other influences such as peer‐social interactions posed as challenges for example, some mother’s reported only buying juice after their child was exposed to it in daycare. Husband’s food preferences and grandparent’s indulgent behaviors were also viewed as a challenge to promoting healthy foods in the home. Understanding how cultural and environmental factors influence parental feeding and may help tailor obesity prevention efforts among Hispanics.

Grant Funding Source: Supported by the University of Rhode Island
Evaluation of dietary patterns and nutritional status of families living in an indigenous population in Chiapas, Mexico (LB469)
Authors: Romano Diana , Guerrero Jenifer , Garcia Olga
Published: June 2021

The objective of this cross‐sectional study was to evaluate the nutritional status and dietary pattern (DP) used by the Special Programme for Food Security (PESA/FAO) in an indigenous population (Chol and Tzeltal) in Chiapas, Mexico. Height, weight, body fat (%) and clinical signs of malnutrition were evaluated in 192 adults and 153 children (<12 y). Family’s DP was measured by collecting food intake data from all family members in a week; calorie, protein, and fat intake were determined. Also, family intake of three food categories was evaluated: 1. Basic: energy providers (cereals); 2. Protectors: vitamin and mineral sources (fruits and vegetables) and 3. Builders: protein suppliers (meat and legumes). Prevalence of <‐2 HAZ z‐score in children was 58%, 29% had <1 HAZ z‐score, 41% had <‐1 WHZ z‐score and 1% were overweight/obese. According to the clinical signs (dry hair, glossitis and hair pigmentation) the main nutrient deficiencies in children were due to lack of energy, protein and iron. Prevalence of overweight/obesity among adults was 44% and 1.5% were underweight. Women had 27.4±7.8% body fat and men had 17.7±6.0%. Families’ DP reported a high intake of basic and protector foods and low intake of protein suppliers. In conclusion, both undernutrition and obesity are highly prevalent in an indigenous population in Chiapas, and public health interventions are urgently needed.
Prenatal breastfeeding confidence is not associated with infant feeding method at one month postpartum (LB479)
Authors: Chezem Je Carol , Bolin Jocelyn
Published: June 2021

Background and Objective: Previous studies have observed a positive relationship between breastfeeding confidence and breastfeeding duration. The objective of this study was to describe associations between prenatal confidence and breastfeeding at one month postpartum.

Methods: Expectant mothers planning to breastfeed for the first month postpartum were recruited from clinics and physicians’ offices. Maternal breastfeeding confidence was assessed in the third trimester of pregnancy using a 10‐item written survey developed by O’Campo et al. Pattern of infant feeding (fully formula fed or partially/fully breastfed) was determined at one month postpartum by telephone interview.

Results: Of the 60 subjects who completed all study requirements, most were White (85%), educated beyond high school (72%) and married (73%). Initially, survey structure and reliability were assessed by exploratory factor analysis and Cronbach’s alpha. Results indicated a unidimensional solution was optimal with a Cronbach’s alpha value of 0.839 indicating good internal consistency. The mean confidence score was 4.6 + 0.82. At one month postpartum, 22 mothers (37%) reported formula‐feeding and 38 (63%) reported breastfeeding their infants. There was no significant difference in confidence score between mothers who formula‐fed and those who breastfed their infants.

Conclusion: Findings from the current study are not in agreement with previous research on breastfeeding confidence. Differences may be due to the limited time frame and/or small sample size in the current study.
Thyroid hormone modulates CaMKII δ levels in cultured neonatal cardiomyocytes (LB48)
Authors: Taniguti Ana Paula , Barreto‐Chaves Maria Luiza
Published: June 2021

Previous studies have implicated that intracellular calcium ion (Ca2+) is a mediator in the regulation of cardiac remodeling. Changes in intracellular Ca2+ concentration regulate the activity of several kinases, including Ca2+/calmodulin dependent kinase II δ (CaMKII δ), the predominant cardiac form involved to hypertrophy. Disorders in Ca2+ homeostasis, cardiac hypertrophy and contractile dysfunction are important features observed in hyperthyroidism, however, the contribution of CaMKII δ in cardiomyocyte growth mediated by thyroid hormones (TH) was not been elucidated yet. In the present study, we evaluated the non‐genomic and genomic effects of TH on CaMKII δ levels in neonatal rat cardiomyocytes. Cardiomyocyte primary cultures (n=3) were obtained from 1‐ to 3‐day‐old Wistar rat by enzymatic disaggregation. At 72 h after platting, the ventricular cardiomyocytes were treated for 5 min, 10 min, 15 min, 30 min or 24 h, with serum‐free medium (control cells) or serum‐free medium containing T3 (triiodothyronine, 10ηM, Sigma Aldrich). CaMKII δ expression of was evaluated by Western blot and normalized to GAPDH expression. Data were analyzed by two‐way ANOVA. We found that T3 induced a significant increase (2 fold) on CaMKII δ expression levels only after 24 h of treatment (p蠄0.05 vs. control), but no effect was observed after rapid treatment with T3. The involvement of this kinase on cardiac hypertrophy induced by TH will be evaluated lately.

Grant Funding Source: Supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Platelet‐activating factor‐receptor in chemotherapy mediated acquired resistance of melanoma (LB495)
Authors: Sahu Ravi , Ocana Jesus , Ferracini Matheus , Konger Raymond , Travers Jeffrey
Published: June 2021

Pro‐oxidative stressors suppress host immunity via generating oxidized lipids with Platelet‐activating Factor‐receptor (PAF‐R) agonist activity. Since many chemotherapeutic agents are potent pro‐oxidative stressors, the present studies investigated if chemotherapy could thwart host anti‐tumor immunity against melanoma via PAF‐R activation. We demonstrate that treatment of melanoma cell lines and tumors with chemotherapeutic agents generates PAF‐R‐agonists. In a dual tumor model, we show that intratumoral chemotherapy with melphalan or etoposide of one tumor significantly augments the growth of other (untreated) tumor in wild‐type but not in PAF‐R‐deficient hosts. Chemotherapy‐mediated PAF‐R‐dependent increased tumor growth is blocked by systemic administration of antioxidants and cyclooxygenase‐2 (COX‐2) inhibitors. In addition, depleting antibodies against regulatory T cells (Tregs) significantly attenuated chemotherapy‐mediated enhanced tumor growth, suggesting the role of Tregs. Moreover, using FoxP3EGFP transgenic mice, we show that COX‐2 inhibitor blocked intratumoral Tregs, indicating that Tregs are downstream to COX‐2. Finally, PAF‐R agonists were identified in perfusates of melanoma patients undergoing isolated limb perfusion with melphalan chemotherapy. These findings provide evidence for a novel pathway by which PAF‐R agonists produced via chemotherapy modulate tumor growth via COX‐2‐Treg pathway. These studies explain some instances of chemotherapy treatment failure and provide potential therapeutic targets that could enhance the overall anti‐tumor effectiveness of chemotherapy.

Grant Funding Source: NIH R01 HL062996, AICR 09A062, ACSIRG 4185607, Showalter Research Trust Fund 4485602
Camel milk and urine inhibits inflammatory angiogenesis in mice via downregulation of proangiogenic and proinflammatory cytokines (LB499)
Authors: Alhaider Abdulqader , Sarwati Sarita , Korashy Hesham
Published: June 2021

Camel milk and urine has traditionally been used to treat cancer, but this practice awaits scientific scrutiny, in particular its role in tumor angiogenesis, the key step involved in tumor growth and metastasis. We aimed to investigate the effects of camel milk and urine on key components of inflammatory angiogenesis in the murine cannulated sponge implant angiogenesis model. Polyester‐polyurethane sponges, used as a framework for fibrovascular tissue growth, were implanted in Swiss albino mice and camel milk and urine (25, 50 and 100 mg/kg/day) was administered for 14 days through installed cannula. The implants collected at day 14 post‐implantation, were processed for the assessment of hemoglobin (Hb), myeloperoxidase (MPO), N‐acetylglucosaminidase (NAG) and collagen, which were used as indices for angiogenesis, neutrophil and macrophage accumulation and extracellular matrix deposition, respectively. Relevant inﬂammatory, angiogenic and ﬁbrogenic cytokines were also determined. Camel milk and urine systemic treatment attenuated the main components of the ﬁbrovascular tissue, wet weight, vascularization (Hb content), macrophage recruitment (NAG activity), collagen deposition and the levels of vascular endothelial growth factor (VEGF), Interleukin (IL)‐1β, IL‐6, IL‐17, tumor Necrosis Factor (TNF)‐α and transforming growth factor (TGF‐β) in a dose dependent manner. Camel milk and urine treatment reduced VEGF expression and microvessel density in vivo. A regulatory function of camel milk and urine on multiple parameters of the main components of inflammatory angiogenesis has been revealed giving insight into the potential therapeutic benefit underlying the anti‐cancer actions of camel urine.
Fipronil induces lung inflammation in a mouse model (LB508)
Authors: Merkowsky Kaitlin , Singh Baljit , Gill J.P.S. , Sethi R.S.
Published: June 2021

Fipronil is an insecticide present in anti‐flea and tick products used on household pets and on crops. Fipronil kills organisms by blocking the GABA‐gated chloride channels. Presently, very little data exists on the pulmonary impact of fipronil on mammals who may be exposed through inhalation, especially in situations where workers do not wear masks during sprays. We used a mouse model to investigate the pulmonary effects of oral or intranasal fipronil (8 mg/kg/bw for 7 days; n=15). Control mice (n=9) were given either ethanol (intranasal) or groundnut oil (oral) for 7 days. Hemotoxylin and eosin stained lung sections showed accumulation of inflammatory cells and changes in lung architecture in both the oral and intranasal groups compared to the controls. To determine whether exposure to fipronil may change susceptibility to bacterial infections through alterations of Toll‐like receptors (TLRs), TLR4 and 9 immunohistochemistry was performed. TLR4 was present in the nucleus of epithelium cells in the oral group and a robust staining observed in the alveolar septum, blood vessel and bronchial epithelium of the intranasal group. Increased TLR9 staining in the alveolar macrophages and septa was observed in the fipronil exposed groups compared to the control groups. We have also tested the effects of fipronil (5.72 um/1 ml) on U937 macrophage cell line, which reduced the viability of these cells. These data suggest that fipronil induces lung inflammation in vivo and cell death in vitro. Furthermore, the increased expression of TLR4 and TLR9 may increase susceptibility for bacterial lung inflammation.
Lung inflammation associated with acute necrotizing pancreatitis in dogs (LB513)
Authors: Vrolyk Vanessa , Wobeser Bruce , Al‐Dissi Ahmad , Carr Anthony , Singh Baljit
Published: June 2021

There is evidence that dogs suffering from acute necrotizing pancreatitis (ANP) can develop respiratory disease. However, there are no data describing the lung histopathology in clinical cases of ANP in dogs. Therefore, we hypothesized that lungs of dogs that died due to ANP (n=21) will be inflamed along with recruitment of PIMs, a pro‐inflammatory cell normally present in ruminants and equines, compared to lungs of control dogs (n=6). Histologic sections of the pancreas graded independently by two blinded pathologists showed the presence of necrosis and infiltration of inflammatory cells (scores 3‐10) whereas control dogs received a score of 0. Lungs from ANP dogs showed no (score of 0/6) to moderate (score 3/6) inflammation, but there was no statistical difference between the lung histological score of the control (median ± SEM: 0.5±0.3) and the ANP (median ± SEM: 1.5±0.2) dogs. However, immunohistochemistry revealed an increase in the numbers of septal macrophages/monocytes in lungs of ANP dogs (median ± SEM: 138±49) compared to control dogs (median ± SEM: 1.5±0,2; p<0,01). There was a moderate correlation (r=0.46) between the recruitment of macrophages/monocytes in the lungs and the total inflammatory cells score in the pancreas of ANP dogs. Only ANP dogs showed expression of vWF in alveolar septal capillaries and septal inflammatory cells. iNOS was expressed only in alveolar macrophages of ANP dogs. TLR4 and IL6 were variably expressed and localized in alveolar macrophages and septal inflammatory cells in lungs from both ANP and control dogs. We conclude that dogs suffering from ANP have pronounced lung inflammation along with recruitment of PIMs and increased expression of vWF. The PIMs may increase the susceptibility of the host to lung inflammation in response to circulating endotoxins.

Grant Funding Source: Supported by Companion Animal Health Fund
Kidney hyperfusion induces immunological reactions in glomeruli (LB525)
Authors: Tang Qunye , Li Zheng , Zhao Zhonghua , Zhang Zhigang , Lu Limin , Zhu Tongyu , Vanhoutte Paul , Shi Yi
Published: June 2021

Objectives. Endothelium is the first line of contact with immune system. It is known that activated lymphocytes impair endothelial functions. However, whether endothelial dysfunction could initiate immune reaction remains unclear.

Methods. C57/BL6 mice were randomly grouped into sham‐operated (2K), 1 kidney removal (1K) and 5/6 nephroectomy groups (5/6K‐off). Mouse glomeruli were collected for polymerase chain reaction (PCR) and immunohistochemistry study.

Results. Four weeks after nephrectomy 5/6K‐off mice had a significantly higher level of renal blood flow, compared with 2K and 1K groups. Urine protein levels, which were normalized with remaining glomeruli, were similar in the three groups. There were no significant increase in levels of macromolecules in urine samples of 5/6K‐off group. Glomerular filtration experiments also supported it. These results suggested that glomeruli of 5/6K‐off group have normal function. In PCR experiments, the expressions of major histocompatibility complex Q lotus, but not the major histocompatibility complex T lotus, were significantly increased in both 1K and 5/6K‐off groups. mRNA expression of both CD80 (B7‐1) and CD86 (B7‐2), costimulatory signals for T cells, were significantly increased in 1K and 5/6K‐off groups. These finding suggests that kidney hyperfusion could initiate immune reaction. Complement component 5a receptor mRNA expression was increased, whereas complement decay‐accelerating factor (also known as CD55) expression was decreased in 5/6K‐off group.

Conclusion. The present study suggests that kidney hyperfusion induces immunological reactions in glomeruli. The complement pathway plays important roles in these reactions. Further experiments will focus to identify the underlying mechanisms.

Grant Funding Source: National Natural Science Foundation of China 81373412
High‐density lipoprotein Mimetic peptide 4F acts on the intestinal microbiota and reduces systemic inflammation (LB533)
Authors: Pourtabatabaei Nasim , Shabihkhani Maryam , Vakili Ladan , Navab Kaveh , Barseghian Zarina , Seddigh Farhad , Hough Greg
Published: June 2021

High‐density lipoprotein (HDL) mimetic peptides have been studied in animals and humans for their ability to improve biomarkers of inflammation and for their ability to decrease atherosclerotic lesions or tumor burden in preclinical studies. The mechanism of action of these peptides is due to their remarkable ability to bind oxidized lipids including ox‐phospholipids, oxidized metabolites of arachidonic and linoleic acids, ox‐sterols and lysophosphatidic acids. Single oral doses of 4.3 and 7.14 mg/kg of the HDL mimetic peptide D‐4F significantly improved the HDL‐anti inflammatory index in patients with coronary heart disease compared with patients that received placebo. The intestine appears to be a major site of the action of the peptide. When Low‐density lipoprotein (LDL) R‐/‐ mice on Western Diet were treated with the peptide orally or by subcutaneous injection, the concentration of the peptide in the enterocytes was similar and plasma superoxide scavenging activity (SAA), lysophosphatidic acids, 5‐Hydroxyeicosatetraenoic acid (5‐HETE), 12‐HETE, 15‐HETE, 9‐Hydroxyoctadecadienoic acid (9‐HODE), 12‐HODE, Prostaglandin D2 (PGD2), PGE2 and Thromboxane B2 (TXB2) were similarly affected. Atherosclerotic lesions were also reduced to a similar extent.
NADPH oxidase inhibitors as regulators of vascular relaxation in aortic rings of diabetic rats with streptozotocin (LB539)
Authors: Macías Martha , Ahedo Dalia , Bobadilla Rosa , Martínez Federico , Mera Elvia , Padilla Itzia , Rosales Martha
Published: June 2021

In diabetes can be present endothelial dysfunction, which is related with the increase in the free radicals production in blood vessels. One of the free radicals present in a great amount is superoxide anion (O2.‐), wich is produced by NADPH oxidase (mainly Nox2). Due to Nox2 could be a therapeutic target in the cardiovascular damage of diabetes. In this research were used Nox2 inhibitors, apocynin and apocynin derivatives two ether and two ester that containing a hydrocarbon chain with different length, to reduce the production of O2.‐, and know if this compound have an effect to relax the vascular smooth muscle (VSM). To evaluate the effect to relax the VSM was using aortic rings of diabetic rats induced with streptozotocine; the results show that the best compounds to relax the aortic rings are compound 1 and 4. Then in order to evaluated if the effect observed in the aortic rings are on the endotelial cells. The production of O2.‐ was measure by the citocromo oxidation in endothelial cells from diabetic rats with and without compounds were the compound 1was better than the others, and reduce the O2.‐ production.Macías‐Pérez et al., Biosciences Report 2013.
The bystander and systematic adverse effects of senescent preadipocytes (LB54)
Authors: Xu Ming , Stout Michael , Pirtskhalava Tamar , Giorgadze Nino , White Thomas , Jensen Michael , LeBrasseur Nathan , Tchkonia Tamar , Kirkland James
Published: June 2021

Introduction: Senescent cells with essentially irreversible loss of replicative potential accumulate in various tissues with aging. Cellular senescence is associated with a pro‐inflammatory senescence‐associated secretory phenotype which the Janus‐associated kinase (JAK) pathway plays a role in. We hypothesized that senescent preadipocytes contribute to age‐related dysfunction of healthy cells.

Methods: Preadipocytes from lean subjects were X‐irradiated to induce senescence. We co‐cultured non‐senescent cells with senescent cells or cultured them in conditioned medium (CM) from senescent preadipocytes to test the inflammation, differentiation and insulin sensitivity. Macrophage migration in response to CM was assayed as well. Moreover, we treated old or obese mice with Ruxolitinib, a JAK1/2 inhibitor, to test the beneficial effect.

Results: The adipogenesis of preadipocytes was impaired by exposure to senescent cells. Macrophage migration was greater in response to senescent preadipocytes. Inflammation was increased in both adipose tissue and preadipocytes exposed to CM from senescent cells. All these adverse effect could be partially rescued by JAK inhibitor. In addition, senescent preadipocytes inhibited insulin‐induced phosphor‐AKT in adipose tissue and HepG2 cells. Insulin‐induced glycogen synthesis was also inhibited in HepG2 cells. Two months of Ruxolitinib treatment reduced inflammation, reversed age‐related lipodystrophy and improved physical function in frail, 24 month old mice. It also improved insulin sensitivity in diet‐induced obese mice.

Conclusion: Senescent preadipocytes contribute to age‐related impairment in adipogenesis, fat tissue inflammation, and insulin resistance.

Grant Funding Source: supported by NIH AG13925 and Robert and Arlene Kogod Center on Aging in Mayo Clinic.
Gene expression of enzymes in the O‐GlcNAcylation pathway in type 2 diabetics vs. non‐diabetics (LB540)
Authors: Gan Kevin , Hsu Tiffany , Ngo Cynthia , Phuong Patrick , Gonzalez, Moises Moises , Suarez Jorge , Fricovsky Eduardo
Published: June 2021

Protein O‐GlcNAcylation has been shown to play a major role in the development of type 2 diabetes and diabetic complications. This study is investigating differences of gene expression of proteins involved in the hexosamine biosynthetic pathway (HBP) in patients using a non‐invasive technique. We aim to compare the relative changes in gene expression of HBP enzymes in type 2 diabetics versus non‐diabetics. We hypothesized that type 2 diabetics have a higher gene expression of these enzymes than non‐diabetics. Patients from the UCSD Free Clinic consented to take part in the study. They were asked to fill out a demographic, current disease states and past family history of diabetes survey. Afterwards, they had their cheek swabbed to collect buccal epithelial cells processed and frozen for analysis. Gene expression was quantified using Real‐Time PCR and the respective enzyme primers. PeriPrimer software determined the relative amount of gene expression and were normalized to the abundance of beta actin mRNA. A two‐group mean comparison t‐test was performed to determine the difference in amount of gene expression between diabetics and non‐diabetic patients with a statistical significance at p < 0.05. Eighty‐two patients participated in the study with fifteen patients who were diabetic. Power was calculated to be 39.51%. The results showed changes in genetic expression of O‐GlcNAc transferase (p <0.05). Despite the small sample size of diabetic patients, and lower power, we observed a significant difference in gene expression between diabetics and non‐diabetics. Potentially these changes in gene expression can be utilized as biomarkers to assess the status of diabetes using a non‐invasive technique.
Hematopoietic effect of the aqueous calyx extract of Hibiscus sabdariffa in rats (LB549)
Authors: Umoren Grace , Elias Simiat , Adeneye Adejuwon
Published: June 2021

Cyclophosphamide, a widely‐used immunosuppressive agent, may adversely affect the bone marrow. This study investigated the hematopoietic effect of the aqueous calyx extract of Hibiscus sabdariffa (HS) in cyclophosphamide‐induced anaemic rats. H. sabdariffa (HS) is used locally as a hematinic concoction supposedly to raise blood level. 50g of pulverized HS leaves were boiled in 500ml distilled water and the filterate air‐dried at 400C. Wistar rats (n = 24) treated as approved by Lagos State University College of Medicine Experimental Animal Ethics Committee, were grouped into 4: Control ‐ water ad libitum; Group I‐ 200mg/Kg/day HS Extract; GP II 30mg/kg/day Cyclophosphamide for 5 days and Group III – HS 200mg/Kg/day for 30 days + 30mg/kg/day Cyclophosphamide for 5 days. Blood sample was collected directly from the heart chambers of the rats after a light anaesthesia with diethyl ether. In Group I, HS extract caused increase in Hb, PCV, RBC, platelet (PLT) count, MCV, and MCH (p<0.05) compared with Control. In Group II, there was significant reduction in Hb, PCV, RBC, MCV, MCH and PLT count (p<0.05) compared with control (p<0.05) and Group I (p<0.05). In GP III, Hb, PCV, RBC, and platelet count were similar to that of Gp I (p > 0.05). The results from this study suggest a hematopoietic potential for the aqueous leaf extract of H. sabdariffa.
U‐bang‐haequi tang, an herbal prescription ameliorates acute inflammation through inhibition of NF‐kB in vivo and in vitro (LB550)
Authors: Kim Sang Chan , Jung Ji Yun , Hwangbo Min , Jee Seon Young , Jegal Kyung Hwan , Park Sang Mi , Cho Il Je , Kim Young Woo
Published: June 2021

Inflammation is a key response of the immune system to infection, and also a complex biological symptom of specific tissues to harmful injury, which is often caused by invading pathogens. U‐bang‐haequi tang (UBT) is an herbal prescription containing Arctii fructus and Forsythia suspensa as its main components. It has been used in traditional Korean medicine to treat a variety of diseases such as fever, infection, and inflammatory disorders. Although UBT has long been used as an herbal remedy, the underlying mechanism of its beneficial effects has not been clearly verified. This research investigated the effects of UBT against an acute phase of inflammation. We measured induction of both nitric oxide (NO) and prostaglandin E2 (PGE2) expression as well as levels of related proteins in a lipopolysaccharide (LPS)‐stimulated macrophage cell line. Further, paw swelling was measured in carrageenan‐treated rats. Protein expression levels were determined by immunoblot analyses, and production of NO and PGE2 was assessed by enzyme‐linked immunosorbent assay. Carrageenan treatment promoted infiltration of inflammatory cells and paw swelling, whereas oral administration of 0.3 or 1 g/kg/day of UBT inhibited the acute inflammatory response. UBT also inhibited mRNA and protein expression levels of iNOS. UBT treatment inhibited NO and PGE2 production, nuclear accumulation of NF‐κB, and elevation of Iκ‐Bα phosphorylation, all of which were induced by LPS. Moreover, UBT treatment significantly blocked the ability of LPS to induce phosphorylation of p38 and c‐Jun NH2‐terminal kinases. In conclusion, UBT prevented both acute inflammation in rats as well as LPS‐induced NO and iNOS gene expression in RAW264.7 cells via inhibition of NF‐κB.

Grant Funding Source: This work was supported by the National Research Foundation of Korea grant funded by the Korea government [MEST](No.2012‐0009400).
Methanol extract of Artemisia apiaceae Herba attenuates acute inflammation via NF‐κB pathway (LB552)
Authors: Cho Il Je , Park Sang Mi , Ryu Ji Cheol , Jee Sun Young , Kim Young Woo , Kim Sang Chan
Published: June 2021

Artemisia apiaceae Herba is one of the most widely used herb for the treatment of malaria, jaundice and dyspeptic complaint in oriental medicine. In the present study, we have investigated the effects of methanol extracts of A. apiaceae Herba (MEAH) on the induction of inducible nitric oxide synthase (iNOS) and pro‐inflammatory mediators by lipopolysaccharide (LPS) in Raw264.7 macrophage cells, and also evaluated the in vivo effects of MEAH on carrageenan‐induced paw edema in rats.

MEAH treatment in Raw264.7 cells significantly decreased LPS‐inducible nitric oxide production and the expression of iNOS in a concentration dependent manner, while MEAH treatment (up to 100 μg/ml) showed no cytotoxic activity.

Results from immunoblot analyses and ELISA revealed that MEAH significantly inhibited the expressions of cyclooxygenase‐2, tumor necrosis factor‐α, interleukine‐1β, interleukine‐6 and prostagrandin E2 in LPS‐stimulated cells. As a plausible molecular mechanism, increased degradation and phosphorylation of inhibitory‐κBα, and nuclear nuclear factor‐κB accumulation by LPS were partly blocked by MEAH treatment. Finally, MEAH treatment decreased the formation of paw edema and infiltration of inflammatory cell induced by carrageenan in rats. These results demonstrate that MEAH has an anti‐inflammatory therapeutic potential, which may result from inhibition of nuclear factor‐κB activation, thereby decreasing the expression of pro‐inflammatory genes.

Grant Funding Source: This work was supported by the National Research Foundation of Korea grant funded by the Korea government [MEST] (No.2012‐0009400).
Pioglitazone Regulation of Glucose in a High‐Salt‐Fed db/db mice. (LB553)
Authors: Obih Patience , Ezebuenyi Michael , Jimoh Akeem
Published: June 2021

Estimates from the World Health Organization indicate that there are some 347 million people diagnosed with diabetes worldwide. Diabetes mellitus is a metabolic disorder characterized by hyperglycemia, and associated with abnormalities in carbohydrate, fat and protein metabolism. Complications of diabetes include microvascular, macrovascular, and neuropathic problems. A recent study showed that more than 30% of people recently diagnosed with type 2 Diabetes had hypertension. Diabetes is also the leading cause of end‐stage‐renal disease which manifests as disturbances in the ability of the kidneys to handle sodium. The thiazolidinediones (TZDs) e.g. pioglitazone introduced for the treatment of type 2 Diabetes are ligands for peroxisome proliferator activated gamma receptors (PPAR gamma). They reduce glucose level by reducing insulin resistance. The objective of this study was to evaluate the impact of high‐salt‐diet on pioglitazone treated db/db mice. Groups of 6 weeks‐old db/db mice obtained from Jackson Laboratory were given either pioglitazone (0.02% ) in diet or pioglitazone (0.02%) plus high salt‐diet (8% NaCl) for twelve weeks. Control groups received either pioglitazone or high salt diet. During the course of treatment, urine volume, urine sodium, protein, creatinine and blood glucose were monitored in the animals. There was a significant reduction in blood glucose in the db/db mice that received pioglitazone. There was also a more significant reduction in blood glucose in the animals that received pioglitazone plus high‐salt diet than those that received only pioglitazone. The mice on a high salt diet excreted more sodium than those on normal food. Does salt play a role in glucose regulation and in the presence of an antidiabetic agent like pioglitazone? From our results, high salt diet seems to play a role in glucose regulation in the presence of pioglitazone in db/db mice. However, more studies are needed to elucidate this observation.

Grant Funding Source: NIH SCORE2