bluuebunny/crossref_metadata_2025 huggingface dataset:
Trends of Waist Circumference Distribution and Abdominal Adiposity among Chinese Adults, 1993‐2004
Authors: Zhang Bing , Wang Huijun , Du Shufa , Zhai Fengying , Popkin Barry M
Published: June 2021

Waist circumference (WC) is a strong predictor of hypertension and other noncommunicable diseases in China. This study examines temporal trends in waist circumference and the prevalence of abdominal adiposity trends among Chinese adults aged 20–65 from 1993 to 2004. We use data of the China Health and Nutrition Survey collected in 1993, 1997, 2000 and 2004. Compared with 1993, the 2004 waist circumference distribution curves flattened and shifted rightwards in males and females. Mean waist circumference increased from 76.8cm to 82.8cm among males and from 75.1cm to 79.3cm among females. A gradient of increasing prevalence of abdominal obesity was also observed in males and females. Similar trends of increasing waist circumference and abdominal obesity were observed in underweight, normal weight, overweight and obesity subjects defined by WHO recommended cutoff points of body mass index (BMI). These results demonstrate the rapid increase in obesity, especially abdominal obesity, among Chinese adults. Primary prevention of abdominal obesity should be a major public health problem in China.
“Nibbling here and there”: The what, when, and why of snacking
Authors: Kinney Rachel Ann , Springer Cary , Raynor Hollie , Spence Marsha , Jahns Lisa
Published: June 2021

Objective: A consistent definition for snacks has not been developed in dietary research despite snacking's significant contribution to overall dietary intake. The purpose of this research was to examine self‐reported definitions of a snack.

Methods: 663 18–19yo entering freshman responded to the question, “How would you define the word “snack”?” Qualitative Analysis was completed using SPSS Text Analysis for Surveys software and manually categorizing responses into themes.

Results: The three largest snack categories were Not a Meal (72%), Small Portion (39%), and Hungry (26%). Twenty‐eight percent of respondents’ snack definitions were counted in two of the three categories and 12% were counted in all three. Additional categories include Types of Food (16%), Not Hungry at Intake (4%), Quick and Easy (3%), and Caloric Amount (1.5%). Not a Meal was less likely to be reported by African Americans than Whites (55% vs. 74%, p<0.01). Small Portion and Hungry were less likely to be reported by males than females (34% vs. 43%, p= 0.02) and (22% vs.30%, p=0.03), respectively. Reported weight status was not related to snack definitions.

Conclusion: Defining a snack appears to have multiple criteria that may be influenced by the participant's gender and race. Providing a clear, universal definition of a snack may be difficult in dietary research.
High and low maternal protein intake of pregnant rats reduces incorporation of food derived amino acids into fetal protein
Authors: Petzke Klaus Jürgen , Daenzer‐Wiedmer Maren , Günther Jeanette , Lang Iris , Kucia Merzena , Klaus Susanne , Metges Cornelia
Published: June 2021

It has been shown that maternal dietary protein (P) intake affects fetal and post weaning growth (Daenzer et al., J. Nutr. 132: 142, 2002). We studied amino acid (AA) metabolism of pregnant mothers and their fetuses. Female rats were pair‐fed isocaloric diets of either low (LP, 10%), adequate (control, CP, 20%) or high casein content (HP, 40%) from start of mating throughout gestation. On gestation d 14 and 19 the plasma AA concentrations and relative incorporation (RI) of orally administered 13C‐lysine (Lys) and 13C‐leucine (Leu) in placental and fetal P based on product/precursor relationships was measured. Placental and fetal weights were not different. Maternal plasma AA showed higher concentrations of branched chain AA in HP, whereas taurine concentrations were increased in LP and CP, but not in HP (P < 0.05). Maternal plasma P 13C‐enrichment decreased with increasing maternal P intake. On d 14 and 19 RILys and RILeu in placental and fetal P were significantly lower (P < 0.05; except P > 0.05 for HP on day 19) in LP and HP fed rats as compared to CP. The data suggest that oxidative losses of AA induced by HP feeding may have consequences for placental and fetal P synthesis. An inadequate maternal P intake alters intrauterine AA metabolism which underlies fetal growth retardation.
The expression of genes involved in amino acid oxidation and gluconeogenesis is down‐regulated by cold exposure in rat liver
Authors: Castro Ana Laura , Palacios Berenice , Torres Nimbe , Tovar Armando R , Aleman Gabriela
Published: June 2021

We have demonstrated that PPAR‐alpha down‐regulates the expression of the histidine degrading enzyme, histidase (HAL), while it up‐regulates enzymes involved in lipid oxidation. PGC‐1 is a well‐known PPAR‐alpha coactivator, that is over‐expressed during cold exposure (CE). The aim of the present work was to study whether the induction of PGC‐1 by CE regulates the expression of genes involved in amino acid catabolism (HAL, tyrosine aminotransferase (TAT)), as well as of genes involved in lipid oxidation and gluconeogenesis (CPT‐1, PEPCK, PPAR?, PGC‐1 alpha and ‐1 beta) in rat liver. Our results show a decreased expression in HAL, TAT and PEPCK after 12 h CE when compared with those rats mantained at room temperature. PGC‐1 alpha, ‐1?, and CPT‐1 expression was increased after CE, with no change in PPAR‐alpha mRNA abundance. Our data suggest that an induction of PGC‐1 alpha and ‐1 beta by CE activates PPAR?, that in turn increased the expression of genes involved in fatty acid oxidation, and at the same time decreased amino acid catabolism for body protein sparing.
Protein requirements in healthy school‐age children determined by using the indicator amino acid oxidation technique
Authors: Elango Rajavel , Humayun Mohammad A , Ball Ronald O , Pencharz Paul B
Published: June 2021

Protein (PRO) requirements in school‐age children were determined by the indicator amino acid oxidation (IAAO) method. Four, healthy children (8 – 11 yr) each randomly received a minimum of three PRO intakes below 0.9 and three intakes above 0.9 g/kg/d; range = 0.16 – 1.84 g/kg/d. The diets were isocaloric and provided energy at 1.7 X REE. PRO was given as an amino acid mixture based on egg protein composition, except phenylalanine which was maintained constant across intakes. PRO requirements were determined by measuring the oxidation of L‐[1‐13C]‐phenylalanine to 13CO2 (F13CO2). Breath and urine samples were collected at baseline and isotopic steady state. Linear regression crossover analysis identified a breakpoint (requirement) at minimal F13CO2 in response to different PRO intakes.

Preliminary results indicate the mean and population‐safe PRO requirements to be 1.35 and 1.59 g/kg/d, respectively. These results are significantly higher than the mean and population‐safe PRO requirements of 0.76 and 0.95 g/kg/d, respectively, currently recommended by the Dietary Reference Intakes (DRI 2005) for macronutrients This study is the first to directly estimate protein requirements in children and suggests that the current recommendations, based on a factorial method are severely underestimated. (CIHR supported)
Methionine splanchnic uptake is increased in critically ill children.
Authors: Sy Jama , Gordon William E , Verbruggen Sascha , Wu Manhong , Castillo Leticia
Published: June 2021

During critical illness the splanchnic area is profoundly affected. There is no information on splanchnic uptake of amino acids in vivo, in critically ill children.

Hypothesis:

Methionine splanchnic uptake in critically ill children will differ from estimates in healthy adults.

Methods:

We studied 24 critically ill children. Eight infants (age 6±3months; wt 6±1.4 kg), eight children (age 5±2 y; wt 14±3 kg and 8 adolescents (15±1.94 y; wt 63.3±21kg), who received continuous enteral feedings, providing protein and energy intake at full maintenance needs. We conducted a simultaneous, intravenous, primed, constant, 9h tracer infusion of L‐[2H3methyl] methionine at 2.5 μmol.kg −1h1 and enteral infusion of L‐[13C] methionine at 5μmol.kg−1h1.

Results:

Plasma methionine fluxes obtained with the enteral 13C and the intravenous 2H3 labels were 85±13 and 30±8, 71±5 and 41±15, and 68±7 and 35±7 μmol.kg−1h−1, respectively for infants, children and adolescents. First pass disappearance of methionine tracer, respectively, was 63, 43 and 47%. The rates of intestinal methionine oxidation were 24±10, 18±6 and 23±8 μmol.kg−1h1

Conclusions:

Methionine splanchnic uptake is higher in critically ill infants when compared to children and adolescents (p<0.05), and higher than values of about 15% reported in healthy adults. NIH DK‐62363, T32 HD‐007445, Ajinomoto 3‐ARP; USDA/ARS 25337387, Sophia Foundation for Scientific Research
Stratifin as a Mediator of Epithelial/Mesenchymal Cross Talking in Skin;
Authors: Ghahary Aziz
Published: June 2021

It is well established that any delays in epithelialization during the process of wound healing, due to either infection or severity of injury, increases the frequency of developing fibrotic conditions. This signifies that in the absence of epithelialization, the ECM continues to accumulate until fibroblasts receive signal(s) from epidermal cells to slow down the dynamic process of healing that leads to maturation and remodelling of the healing wound. Here, we provide a compelling evidence that: 1) there exist a cross‐talking between epithelial and fibroblasts using a co‐culture system, 2) Keratinocytes release two sets of ECM modulating factors that are important in controlling the healing process, 3) One set of this factors, namely stratifin, increases the expression of MMP‐1, 3, 8 and 24 in fibroblasts, 4) the other set reduces the expression of type I and type III collagen in fibroblasts, 5) On the hand, fibroblasts release some unknown factors that control the expression of keratinocyte derived anti‐fibrogenic factors (KDAF). Finally the mechanism by which these factors are released from keratinocytes will also be discussed.

In conclusion, Keratinocyte releasable factors upon epithelialization may function as stop signals for matrix production by fibroblasts and that would slow down the dynamic process of healing wound.
Zinc and Lead Hair Levels in Children Ages 6‐60 Months Living in Three Refugee Camps along the Thailand/Burma Border
Authors: Kemmer Teresa M , Geisler Karen , Reeves Philip G
Published: June 2021

Long term subsistence on refugee rations, with minimal amounts of animal protein, put children at increased risk for zinc (Zn) deficiency. In addition, lead (Pb) levels have never been reported in refugee children. The study objectives were to determine 1) the prevalence of Zn deficiency, 2) associations with Zn deficiency and 3) prevalence of elevated Pb levels.

Using 2‐stage cluster sampling, hair samples from 377 children ages 6–60 months living in Bon Don Yang, Mae La, and Tham Hin refugee camps were obtained and analyzed. Zinc deficiency was defined as Zn levels <70 μg/g and Zn inadequacy as levels between 70–100 μg/g. A very high Pb level was defined as >25 μg/g.

Prevalence of Zn deficiency was 17.9 % and Zn inadequacy was 17.6%. Mean Zn was 136.19±75.95 μg/g. 100% of the children had a very high Pb level (Mean 143.97±35.69 μg/g). By using logistic regression and controlling for age and gender, Zn < 100 μg/g was associated with female gender (P< 0.01), mothers with some education (P< 0.01), lived in camp > 1 yr (P<0.05) and increasing Pb levels (P<0.001).

Hair analyses confirmed that the children had a high prevalence of Zn deficiency and extremely high Pb levels. Rations containing adequate Zn are indicated and sources of Pb should be evaluated. A fortified blended food with Zn (10.0 mg/100g) has since been added to the ration; therefore, a second analysis should be conducted to determine the impact of the fortified ration.
Body Composition by DXA in pre‐school children
Authors: Rodriguez Patricia Noemi , Bermudez Enrique Feliz , Macri Elisa Vanesa , Ponce Graciela , Zeni Susana Noemi , Exeni Ramon Alfonso , Lifshitz Fima , Friedman Silvia Maria
Published: June 2021

Measurements of body composition in children is becoming more widespread. Many technologies are available, but DXA's use is increasing in the last years due to its accuracy. The current obesity epidemic lead us to studied the relationship between body fat mass and bone, due to some evidence that in adolescents and young adults adipose tissue is not beneficial to bone structure.

Objective: To compare body fat mass % (BF%) and bone mineral content % (BMC%) in boys and girls aged 3 to 5. Eigthy five children (males‐ M=40, females‐F=45) from La Matanza, Province of Buenos Aires, Argentina, attending a day care center, were included. There were divided in 6 groups according to weight/length Z scores (WLZ) and gender: 1M and 1F) −1.5 to 1.5, 2M and 2F) 1.5 to 2.0 and 3M and 3F) >2.0 WLZ. BF% and BMC% were measured by Lunar DPX‐L, pediatric software.

Results: BF% and BMC% in: 1M were 18.4±4.6 and 37.7±7.0, 2M 27.9±7.7 and 38.4±6.2, 3M 30.6±6.2 and 34.1 ±4.8, 1F 25.6±5.0 and 41.1±5.2, 2F 35.3±5.7 and 35.7±4.5, and 3F 34.1±7.3 and 401.0±6.2, respectively.

Conclusion: these results suggest that BMC in children aged 3 to 5 remained unchanged, regardless of high BF%. Awarded by UBACyT 004‐003.
Does hunger bias the estimation of food size and food weight?
Authors: Wansink Brian , Vartanian Lenny , Payne Collin R. , Brasel S. Adam , Abram Jenica K.
Published: June 2021

The objective of this study is to understand better if hunger biases size and weight judgments of foods. Participants were asked to estimate the size and weight of five different non‐food items—to serve as a control—and seventeen food items before or after they ate lunch. Surprisingly, regardless of whether participants had lunch or simply rated themselves as being hungry, size and weight estimations remained constant. However, stable individual differences suggest that those participants who were innately more biased—regardless of hunger—when estimating size and weight of non‐food items were also more biased when estimating the same dimensions of food. These results suggest an innate bias for estimating foods, which could affect food consumption patterns.
Low dietary quality is related to high blood lead concentration in school aged children
Authors: Kwon J.I. , Kang D.M. , Kim S.Y. , Sa K.J. , Lee C.G. , Lim J.H. , Jee Y.K. , Ha M.N. , Hong S.J. , Hong Y.C. , Kwon H.J. , Oh S.Y.
Published: June 2021

Lead is a toxic metal widely spread in the environment with bioaccumulative features that raises public health concern. Diet might be associated with blood lead concentration in children. In order to evaluate the association between dietary intake and blood lead concentration, a cross‐sectional study was conducted on 2,050 children aged 7 to 8 years residing in urban areas in Korea while assessing dietary intake by a semi‐quantitative food frequency questionnaire and blood lead concentration by the graphite furnace atomic absorption spectrophotometer. Children were divided into high (n=617) and low (n=1,433) lead groups using a 2 μg/dl cutoff point. In multivariate logistic regression analyses, higher intakes of vegetable protein and iron, phosphorous, potassium, zinc, folic acid and vitamin E were related to lower blood lead concentration. Blood lead concentration was associated positively with intakes of white rice, ramyun, salted vegetables, ice cream and soft drink, and negatively with mixed rice, spinach, seaweed, fruits, and white milk consumptions. These findings suggest low dietary quality may be a risk factor for high blood lead concentration in children. (Supported by a grant from the Ministry of Environment, Korea in 2006)
Branched‐Chain‐Aminoacids content in food and nutritional supplements
Authors: Pereira Paula Cardoso , Vicente Ana Filipa , Cabrita António Silvério
Published: June 2021

Exercising individuals need more protein that sedentary people and should make their efforts to choose high quality protein sources. An attempt should be made to achieve protein needs from whole foods but nutritional supplements, mainly protein powders, can be useful in crucial timings like after physical effort when recovery is first priority. Branched chain aminoacids (BCAA's) are one of the most important criteria to choose a protein source, because they are an important fuel to muscle cells and because of their role in delaying central fatigue.

In the present study we compared different protein sources and evaluated protein and BCAA's content. The results showed that whey protein powders have more protein in one 30g serving that the same amount of beef, chicken and pork lean cuts, tuna, egg whites and whole eggs. And these nutritional supplements are richer in BCAA's and stand close to 2:1:1 ratio of leucine, isoleucine and valine.

A balanced diet can be achieved if the athlete uses whole foods most of the time, but when in a crucial moment to recover or improve performance, protein powders can be more efficient and more convenient that normal foods that have to be cooked and can have other nutrients that delay protein and aminoacids absorption and utilization by muscle cells.
Metabolic Syndrome, Type 2 Diabetes and Controls: Comparisons and Contrast
Authors: Hubbard Richard W. , Westengard James , Guthrie George
Published: June 2021

These metabolic comparisons were from a highly health motivated, self‐selected cohort (ages16–88) composed of four definable categories: Metabolic Syndrome (MS) (n=67), Type 1 (n=16) and Type 2 (n=155) Diabetics and Controls (n=123). The biological assays were performed at Quest‐Nichols Diagnostics. Statistical analysis was performed by one‐way ANOVA, with Bonferroni adjusted probabilities for multiple comparisons (Systat ver.10; SPSS Inc., 2000).

The MS subjects show significantly higher Chol/HDL ratio (p= 0.028) and diastolic blood pressure (0.0004) while the Type 2 diabetics had significantly higher Hbg A1C (p< 0.00005) and higher fasting blood sugar (p<0.00005) values. The MS group and the Type 2 diabetics move together with respect to the Control group for Vitamin D deficiency (p = 0.00014), and with increased triglycerides (p<0.00005), and decreased HDL (p<0.00005), while there are also significantly increased BMI (p<0.00005), CRP (p=0.04326) and systolic BP (p=0.0003) values compared to the control group. New findings from the report by Reis, JP et al, (Diabetes Care 30 (21007) 1549) on hyperparathyroidism in MS, where this does not appear to be the case in Type 2 diabetics (Taylor WH. Khakeeli Dia Met Res Rev 17(3) (2001) 175. Additional studies, relative to weight control, vitamin D deficiency and normalizing parathyroid function in MS will be necessary to clarify this important issue.
Diallyl sulphide protects against N‐nitrosodiethylamine induced liver tumerogenesis
Authors: Nassar Noha , Ibrahim Safinaz
Published: June 2021

Diallyl sulfide (DAS), a major constituent of garlic, has been documented as a potent inhibitor of tumerogenesis in various animal models. This study investigated the protective effect of DAS against N‐nitrosodiethylamine (NDEA) induced liver tumerogenesis. To this end, two groups of male Sprague Dawley rats (n= 12; 170–200 g each group) received either NDEA (10mg/kg/day for 8 weeks followed by 20 mg/kg/day for the next 6 weeks) to induce liver cancer or NDEA + DAS (200mg/kg/day) for all 14 weeks. Liver lactate/pyruvate ratio, lactate dehydrogenase and glucose‐6 phosphate dehydrogenase activities were measured (energy metabolism indices). Liver malondialdehyde, cytochrome oxidase activities (membrane disintegration biomarkers), DNA level and glutathione S‐transferase activity (DNA fragmentation indices) were also measured. Serum alkaline phosphatase and gama‐glutamyl transferase activities were investigated (liver function). Results showed that administration of NDEA significantly disturbed liver functions as well as most of the measured parameters. Treatment with DAS protected most of the measured liver parameters against the drastic damaging effects caused by NDEA. In summary, we provide evidence that DAS exerts a protective role on liver functions and tissue integrity in face of enhanced tumerogenesis caused by NDEA.

This research is supported by Cairo University, Egypt
Circulating fatty acids and prostate cancer risk in a nested case‐control study: The Multiethnic Cohort
Authors: Park Song‐Yi , Wilkens Lynne R. , Henning Susanne M. , Henderson Brian E. , Kolonel Laurence N.
Published: June 2021

Dietary fat, including specific fatty acids (FAs), has been proposed to contribute to prostate cancer pathogenesis, but findings from studies based on biomarkers have been conflicting. We examined the association between erythrocyte membrane FA composition and prostate cancer risk in a nested case‐control study within a large cohort of African Americans, Native Hawaiians, Japanese Americans, Latinos, and Whites. Analyses included 376 cases and 729 matched controls. Conditional logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs) with adjustment for the matching criteria of ethnicity, age and date of blood draw, and fasting status, as well as family history of prostate cancer, BMI, physical activity, and education. No significant association was found for saturated, mono‐ and polyunsaturated FAs, or for specific n‐3 and n‐6 FAs, even when the analysis was limited to advanced or high grade prostate cancer. An ethnic‐specific analysis of African Americans, the only group with enough case numbers, showed a suggestive, but not statistically significant inverse association between α‐linolenic acid and prostate cancer risk (OR for the highest vs. lowest tertile = 0.39, 95% CI = 0.14–1.09, Ptrend = 0.09). Our findings do not support an association between FAs and prostate cancer risk. Grant sponsor: National Cancer Institute; Grant No. R37 CA54281.
Blueberry consumption inhibits gastrointestinal tumorigenesis in AOM‐treated rats
Authors: Simmen Frank A , Xiao Rijin , Wu Xianli , Prior Ronald L
Published: June 2021

Epidemiological studies show inverse or no association of fruit consumption with relative risk for colorectal cancers. Such studies did not examine specific fruits and in particular, those enriched in polyphenols with high antioxidant capacities. We examined blueberry (BB), a fruit rich in anthocyanins and proanthocyanidins, for effects on gastrointestinal cancers in AOM‐treated Sprague‐Dawley rats. AIN93G diets containing no or 10% freeze‐dried BB were fed to rats over their lifetime. Rats received AOM at days 53 and 60 of age, with colon aberrant crypt foci (ACF) and gastrointestinal tumors quantified 6 and 17 weeks later, respectively. BB decreased ACF (by 24%, P=0.093) in males and increased ACF (31%, P=0.064) in females. Tumor incidence was reduced by BB in mid colon and proximal small intestine of male rats, resulting in a lower (26%, P=0.081) overall intestinal tumor incidence. In all intestine regions, female rats had reduced tumor incidence (∼70%, P<0.001) relative to control males, with no effects of BB manifested. Females had more adenomas (as a proportion of total tumor number) than did males. BB favored (P<0.05) greater adenoma/total tumor ratio in females. Results indicate inhibitory effects of BB on tumor initiation in AOM‐treated male rats and adenoma to adenocarcinoma transition in female rats, thus warranting further studies of BB on colorectal cancer. Supported by CRIS 6251‐51000‐005‐03S
Integration of Embryology and Molecular Biology
Authors: Schoenwolf Gary C.
Published: June 2021

To understand how structural birth defects arise in humans, it is essential to have not only a detailed knowledge of descriptive embryology, but also a comprehensive knowledge of the molecular genetic mechanisms underlying embryonic development. The understanding of birth defects provides a practical reason for the medical or dental student to learn human embryology. To understand the molecular genetic basis of embryology, the student must understand the basic principles of cell and molecular biology and must learn the relevant signaling pathways, animal models, and experimental approaches used in developmental biology. Some examples of how this material is taught to medical and dental students in human embryology courses to gain a molecular genetic understanding of normal and abnormal development will be presented. For disclosure purposes, Dr. Schoenwolf is the senior author of the 4th Edition of Larsen's Human Embryology. Examples will be chosen from this text.
Evaluation of the prevalence of Folate Associated Megaloblastic Anemia among Pregnant Females in Alexandria's Maternal Health Centers in Egypt
Authors: WALY MOSTAFA IBRAHIM‐AHMED , AHMED REEM MHAMED , HILMY MYRIAM ABOU‐SEIF , DARWISH OLFAT ABD‐ELHAMID
Published: June 2021

Megaloblastic anemia is a complication of folate deficiency, and pregnancy precipitates the risk of this anemia due to increased requirements needed for maternal tissue and fetal growth. It is a manifestation of defective DNA synthesis in hematopoietic cells characterized by the presence of large, immature, and dysfunctional red blood cells. Folate deficiency is a primary cause for pregnancy loss and neural tube defect for the pregnancy outcome. In Egypt there is no national data that describes the prevalence of folate deficiency associated anemia among pregnant women. A random sample of 5000 pregnant women, in the second trimester, attending Alexandria's maternal and child health centers were screened for anemia and its types. Our results indicate that 0.1% of the studied population was megaloplastic anemic subjects, characterized by macrocytic anemia with a hemoglobin level (Hb) below 7.0 g/dL and average serum folate level of (2.4±0.4) μg/L, also their recommended dietary allowances (RDA) was low for folate (277μg/day) and vitamin B12 (1.9μg/day). Three months of folate supplementation and dietary counseling educational programs for these cases improved their serum folate level to (3.65±0.2) μg/L as well as their RDA for folate and vitamin B12, indicating the efficiency of these intervention programs for the treatment of megaloblastic anemia in this vulnerable group.
Effect of ozone boluses on the lung function of smokers
Authors: Bates Melissa Lowe , Brenza Timothy M. , Ben‐Jebria Abdellaziz , Ultman James S.
Published: June 2021

During continuous exposure to ozone (O3), smokers have failed to experience the decrements in forced spirometric parameters that have been well‐established for comparable non‐smokers. We hypothesized that these differences in response are due to differences in the longitudinal distribution of O3 uptake. We measured the longitudinal distribution of O3 uptake using the bolus inhalation method but found no difference between populations of 29 non‐smokers (17M, 12F) and 30 smokers (19M, 11F, 4±4 pack‐years). We also measured FEV1 by spirometry and the normalized slope of the alveolar plateau (SN) by capnography. Smokers experienced a significant change in SN (8.1 ± 3.2%, p=0.02) and a small, but significant change in FEV1 (−1.3 ± 0.6%, p=0.03) after inhaling between 75 and 120 20‐mL O3 boluses with an average concentration of 1 ppm. Nonsmokers did not exhibit such responses. We conclude that during a transient exposure to O3, smokers may be differentially responsive to O3 in spite of the similarity of their O3 uptake distribution to that of non‐smokers.
Exposure to a polluted urban environment and to chronic low doses of lipopolysaccharides produces neuroinflammation in mice, and is ameliorated by dark chocolate
Authors: Villarreal‐Calderon Rafael , Osnaya Norma , Garcia Raquel , Stone Ida , Calderon‐Garciduenas Lilian , Reed William
Published: June 2021

Background: Air pollution is a serious environmental problem. Mexico City (MC) children exhibit upregulation of COX2, and IL1beta in the olfactory bulb, frontal cortex, SNC and vagus nerves

Objectives: To test the hypothesis in Balb‐c mice that exposure to a polluted atmosphere and to low doses of LPS IP results in neuroinflammation, and that inflammation could be ameliorated by dark chocolate

Methods: Balb‐c mice age 21 d: A. clean environment, no Rx, B. outdoor/indoor environment in SW or NWMC, C. SWMC mice Rx with dark chocolate 2g/9.5 mg polyphenols/3 times per week, D. lyophilized E coli 0111:B4, 10 muL 750 or 2250 EU/mL, IP/2 /week clean environment. RT‐PCR: COX2 and IL1beta

Results: At 4 m the LPS 6x group had higher frontal levels of COX2 v the control (0.008) and there were significant differences in IL1beta between NW and SW mice in olfactory bulb and frontal (0.006) and level X (0.0001). The OB in the SWMC chocolate group had significantly lower levels of COX‐2 and IL1beta (0.03) compared to the untreated group. At 8m chocolate reduced COX2 in SW mice OB and frontal regions (0.01, 0.03), there was a significant X upregulation of COX2 in NW v SW mice(0.001), and of IL1beta and COX2 in LPS 2x mice (0.01,0.05).

Conclusions: Neuroinflammation in target areas for Alzheimer and Parkinson is present in mice exposed to a polluted urban atmosphere, and in LPS IP exposed animals. Dark chocolate ameliorates neuroinflammation
Is 3D MR molecular imaging of neovasculature prognostic of tumor response to antiangiogenic therapy?
Authors: Caruthers Shelton D , Schmieder Anne , Winter Patrick , Williams Todd A , Allen John S , Hu Grace , Zhang Huiying , Wickline Samuel , Lanza Gregory
Published: June 2021

αvβ3—targeted fumagillin nanoparticles suppress the neovasculature and inhibit tumor growth in Vx2 tumors. The objective of this study was to use MR molecular imaging and 3D neovascular mapping tools to quantify antiangiogenic and tumor growth responses to integrin‐targeted fumagillin nanoparticles in a xenograft tumor model.

Methods:

Nude mice implanted with MDA435 cells received ανβ3‐targeted fumagillin nanoparticles, α5β1‐targeted fumagillin nanoparticles, or saline on days 7, 11, 15, and 19 post implantation. On day 22, tumor volume and neovascularity were assessed using integrin‐targeted MR nanoparticles.

Results:

Neovascularity was sparser in the xenograft model than observed in Vx2 rabbits. α5β1–fumagillin‐nanoparticles decreased (p<0.05) angiogenesis relative to control (1.3±0.3%) to an almost negligible level (0.5±0.3 %) while αvβ3–fumagillin‐nanoparticles did not decrease (p>0.05) angiogenesis (1.0±0.3%, Fig). Neither treatment regimen affected tumor volume.

Conclusion:

Tumors expressing low levels of neovascularity have poor tumor growth response to antiangiogenic treatment. Nascent cancers responsive to antiangiogenic therapy may be differentiated with noninvasive high‐resolution MR molecular imaging, which may be clinically relevant in establishing personalized therapy regimens. Support NIH/NCI CA 119342
Ontogeny of Toll Like Receptor 2 in the Developing Rat Lung
Authors: Benjamin John T , Chaplain Jennifer , Bochkareva Aleksandra , Gebb Sarah
Published: June 2021

Background: Antenatal bacterial colonization and subsequent inflammation of the fetal membranes increases the risk of Bronchopulmonary Dysplasia (BPD), a disease associated with recruitment of various inflammatory cells to the lung. Innate immune signaling through Toll like receptor 2 (TLR2) is used by various pathogens causing antenatal infections. An increase in TLR2 postnatally may thus provide a molecular mechanism by which antenatal bacterial colonization causes subsequent preterm lung disease. Objective: We hypothesized that TLR2 expression increases postnatally in the developing lung, in part because of changes in the oxygen environment. To test this we used a rat model of fetal lung development. Methods: Lungs isolated from embryonic (E) day 15, 16, 18, 20 and postnatal (P) day 2 and 4 rats were used to quantify TLR2 gene expression. For invitro studies, E15 lung explants and mesenchymal cells were isolated and cultured in 21% (postnatal) O2 or 3% (fetal) O2. TLR2 gene expression was measured by RT‐PCR. Results: Lung TLR2 expression increased 700% from E15 to P4 (P < 0.05). Almost half of this increase (300%) occurred in the first 2 days after birth. There was a trend towards increased expression of TLR2 in both explants (243% increase; P = 0.06) and isolated lung mesenchymal cells (244% increase; P = 0.08) cultured in 21% O2 Vs 3% O2. Conclusion: These data suggests that TLR2 gene expression increases in the immediate postnatal period. This postnatal increase may be mediated in part by changes in oxygen environment after birth. In the presence of antenatal bacterial colonization, increased TLR2 postnatally could lead to increased inflammation and subsequent chronic lung injury.
Synthetic nanoparticle vehicles deliver siRNA to vascular endothelial cells for VCAM‐1 knockdown
Authors: Kaneda Megan M , Milbrandt Jeffrey D , Lanza Gregory M , Wickline Samuel A
Published: June 2021

The therapeutic promise of siRNA relies on its ability to knock down genes with minimal side effects. Upregulation of inflammatory genes such as vascular cell adhesion molecule‐1 (VCAM‐1) in atherosclerosis suggests that targeted delivery of siRNA as an antiatherosclerotic therapy might modify disease progression. We have shown previously that liquid perfluorocarbon nanoparticle emulsions (PFC‐NP) can serve as molecularly targeted vehicles for imaging and drug delivery in vivo. In this study, we sought to utilize PFC NP as carriers of siRNA to endothelial cells. Cationic PFC‐NP emulsions were produced with 1,2‐Dioleoyl‐3‐Trimethylammonium‐Propane (DOTAP). siRNA to VCAM‐1 was complexed to the emulsions and incubated with mouse 2F‐2B endothelial cells. We observed that fluorescently labeled siRNA in cells 24 h after transfection as determined by confocal microscopy. Optimized transfection conditions (10 pM PFC‐NP with 5 nM siRNA) produce a 52% decrease in VCAM‐1 mRNA levels after 48h. Corresponding viability data measured 24 h after transfection using flow cytometry indicated 80–90% viability in 2F‐2B cells under similar transfection conditions. Accordingly, these particles offer an efficient method for siRNA delivery to endothelial cells and they can simultaneously serve as a vehicle for molecular imaging to confirm siRNA delivery. This work was supported by NIH grant #HL073646
Observational studies of Dopamine D2‐family agonists in squirrel monkeys
Authors: Desai Rajeev I , Connolly Jennifer , Bergman Jack
Published: June 2021

Dopamine D2‐family (D2) agonists and antagonists produce dose‐related increases in, respectively, scratching and catalepsy‐associated behavior (CAB) in squirrel monkeys.

The effects of D2 ligands on observable behavior were further compared in the present study. Results show that D2 full agonists [(+)‐PHNO, R(−)‐NPA, quinelorane, PD 128907 and 7‐OH‐DPAT] produced dose‐dependent increases in scratching, with effects that varied in magnitude among drugs [R(−)‐NPA ≈ (+)‐PHNO (>35% of session) > quinelorane ≈ PD 128907 ≈ 7‐OH‐DPAT (<25%)]. D2 partial or mixed‐action agonists [terguride (TER), preclamol (PREC), talipexole (TAL), roxindole (ROX), aripiprazole (ARIP)] did not significantly increase scratching. Results also show that, like haloperidol (HAL), TAL, TER, ARIP and ROX produced dose‐related increases in CAB. HAL, TAL, and TER produced high levels of CAB (65–99% of session), whereas the mixed‐action D2 partial agonists ROX and ARIP produced lesser levels of CAB (<40%). The D2 full agonists (+)‐PHNO, 7‐OH‐DPAT and PD 128907 produced no evidence of CAB. Although the roles of D2 subtypes in agonist‐induced scratching or of intrinsic activity in CAB are not well understood, these behavioral endpoints serve as quantifiable measures of D2 receptor activation or blockade in primate species. (supported by NIH/NIDA RO1‐03774, RO1‐10566)
Neither donepezil nor d‐amphetamine improve working memory in the Ts65Dn mouse, a murine model of Down syndrome
Authors: Whitney Katharine , Wenger Galen R.
Published: June 2021

The Ts65Dn mouse, a model of Down syndrome, has a partial trisomy of chromosome 16. Previous research using an incremental repeated acquisition schedule of reinforcement has shown that there is a significant learning impairment in Ts65Dn mice compared to littermate controls (LC) (Wenger et al. Behav. Genet. 34: 105–119, 2004). The purpose of this study was to examine the effect of d‐amphetamine (0.1 – 10 mg/kg) and donepezil (0.3 – 5.6 mg/kg) on working memory in young adult, male Ts65Dn and LC mice responding under a titrating, delayed matching‐to‐position schedule of reinforcement. When the behavior stabilized, the length of the mean delay for the session achieved by the two lines of mice was indistinguishable, but rates of responding engendered by the sample stimulus were higher in the LC mice than in the Ts65Dn mice. Low to moderate doses of donepezil and d‐amphetamine failed to alter the rate of responding or the length of the mean delay for the session. However, significant decreases in the rate of responding and the mean delay length were observed at high doses of both drugs. These results show that the acute administration of these agents is ineffective in improving working memory in either the young adult Ts65Dn mice or age‐matched LC mice. Future studies will evaluate the efficacy of these agents as the animals age and following chronic administration. Supported by NIH grant HD047656 (G.R.W.)
Rab2 and Rab6 differentially regulate anterograde transport and signaling of G protein‐coupled receptors
Authors: Dong Chunmin , Wu Guangyu
Published: June 2021

Rab1, Rab2 and Rab6 are involved in protein transport between the endoplasmic reticulum (ER) and the Golgi. Whereas Rab1 regulates the anterograde ER‐to‐Golgi transport, Rab2 and Rab6 coordinate the retrograde Golgi‐to‐ER transport. We have previously shown that Rab1 selectively modulates the export of G protein‐coupled receptors (GPCRs) to the plasma membrane. In this report, we demonstrated that expression of the GTP‐bound mutant Rab2Q65L significantly attenuated the cell‐surface expression of both α2B‐AR and β2‐AR, whereas the GTP‐bound mutant Rab6Q72L selectively inhibited the transport of β2‐AR, but not α2B‐AR. Similar results were obtained by siRNA‐mediated selective knockdown of Rab2 and Rab6. Consistently, Rab2Q65L and Rab2 siRNA inhibited α2B‐AR and β2‐AR signaling, whereas Rab6Q72L and Rab6 siRNA reduced signaling of β2‐AR, but not α2B‐AR. Similar to β2‐AR, AT1R expression at the cell surface and signaling were inhibited by Rab6Q72L. Furthermore, the nucleotide‐free mutant Rab6N126I selectively attenuated the cell‐surface expression of β2‐AR and AT1R, but not α2B‐AR. These data demonstrate that Rab2 and Rab6 differentially influence anterograde transport and signaling of GPCRs and that Rab6‐coordinated retrograde transport selectively modulates intracellular trafficking and signaling of GPCRs(R01GM076167).
Calcineurin Binding to the mAKAPβ Signalosome
Authors: Kapiloff Michael Seth , Lopez Johanna , Negro Alejandra , Henson Edward , Dodge‐Kafka Kimberly , Bauman Andrea
Published: June 2021

mAKAPβ is the scaffold for a signalosome located on the cardiac myocyte nuclear envelope which is believed to transduce local cAMP and calcium ion signals and to mediate crosstalk with MAP‐kinase signaling pathways. We and others have shown that mAKAPβ binds a set of signaling enzymes that includes protein kinase A, the calcium‐release channel ryanodine receptor, type 4D3 phosphodiesterase, ERK5 and MEK5, and the cAMP‐dependent rap1‐guanine exchange factor epac1. We have recently found that the calcium/calmodulin‐dependent phosphatase calcineurin binds mAKAPβ. Calcineurin is a mediator of cardiac hypertrophy in vivo and in vitro and is responsible for the dephosphorylation and activation of NFATc transcription factors. We have found that mAKAPβ expression is required for the nuclear translocation of NFATc in myocytes and for the induction of myocyte hypertrophy by adrenergic agonists. We now show that the catalytic A‐subunit of calcineurin can directly bind mAKAPβ. This binding is enhanced by calcium/calmodulin in vitro and is induced by norepinephrine treatment of cultured myocytes. Interestingly, the calcineurin binding site on mAKAPβ does not contain a consensus binding site, suggesting a novel calcineurin binding domain. We propose that by binding calcineurin, the mAKAPβ signalosome participates in the regulation of NFATc transcription factors involved in the induction of myocyte hypertrophy.
Concentration dependent contractile effect of methylene blue in human internal mammary artery.
Authors: Seyrek Melik , Ulusoy Hasan Basri , Yildiz Oguzhan , Gul Husamettin , Yildirim Vedat
Published: June 2021

Objective: Methylene blue (MB), an inhibitor of soluble guanylate cyclase, may a lifesaving drug which improves hemodynamic parameters in vasoplegic syndrome. We investigated the relation between MB concentration and vascular responses to clarify the rationale of its use in vasoplegic syndrome.

Methods. The study was performed in organ bath in human isolated internal mammary artery (IMA). The responses were recorded isometrically by a force‐displacement transducer.

Results: MB (10 nM‐100 μM) produced concentration‐dependent contraction in the arteries. The maximal contraction to MB was 44.2% ± 3.8 of 45 mM KCl maximum contraction. pEC50 value was 5.5 ± 0.1. MB caused an insignificant leftward shift of the concentration‐response curve of norepinephrine. Acetylcholine‐induced relaxation in submaximal contracted rings with phenylephrine was recovered nearly six hours later than MB challenge.

Conclusions: We showed that MB caused concentration‐dependent contraction in human IMA. MB inhibited ACh‐induced relaxation during six hours considering muscarinic receptor occupation. The contractile effect of MB is predominantly related to its own effect which proves a rationale for its use in cathecolamine refractory vasoplegic syndrome.
Angiotensin 1–7 decreases nerve stimulated Norepinephrine (NE) and Neuropeptide Y (NPY) overflow from the mesenteric arterial beds of SHR
Authors: Byku Mirnela , Macarthur Heather , Westfall Thomas C
Published: June 2021

NPY is a cotransmitter with NE and ATP in sympathetic nerves. There is evidence for increased activity of the sympathetic nervous system and the renin‐angiotensin system (RAS) as well as a role for NPY in the development of hypertension in experimental animal models and in humans. Angiotensin II (Ang II) is known to facilitate sympathetic neurotransmission; an effect greater in Spontaneously Hypertensive Rats (SHR) than normotensive Wistar‐Kyoto (WKY) rats. A newly discovered product of the RAS is the peptide Angiotensin 1–7 (Ang 1–7). There is evidence suggesting that Ang 1–7 opposes the actions of Ang II resulting in hypotensive effects. The objective of this study was to investigate the role of Ang 1–7 on the nerve stimulated overflow of NE and NPY from the mesenteric arterial bed of SHR and the receptors mediating any effects produced.

Ang 1–7 (0.001, 0.01, 0.1μM) decreased nerve stimulated NE and NPY overflow, as well as perfusion pressure in preparations obtained from SHR. This effect was greater in preparations of SHR than WKY controls. Administration of the Mas Receptor Antagonist (MRA), D‐Ala Ang 1–7, and the Angiotensin Type 2 Receptor Antagonist (AT2A), PD12339 attenuated this effect.

Ang 1–7 decreases the nerve stimulated overflow of NE and NPY in preparations of SHR, while Ang II enhances it. Therefore, Ang 1–7 may counteract the effects of Ang II by acting on AT2 and Mas Receptors. (Sponsored by NIH HL60260 and AHA)
Increased expression of components of the Rho‐A/Rho‐kinase pathway does not compensate for its impaired activation in small mesenteric arteries from endotoxemic rats
Authors: Santos José Eduardo da Silva , Chiao Chin‐Wei , Leite Romulo , Webb R. Clinton
Published: June 2021

Background:

Hyporeactivity to vasoconstrictors is a hallmark of septic shock. Although activation of the Rho‐A/Rho‐kinase pathway has been shown as crucial for development of vascular contraction its role or involvement in septic shock is unknown.

Methods:

We evaluated the relaxation induced by the Rho‐kinase inhibitor Y‐27632 (1 nM to 30 μM) in first order mesenteric arteries from rats treated with lipopolysaccharide (LPS, 10 mg/kg, i.p.) for six or 24 h (or sterile saline; control group). Results: Relaxation induced by Y‐27632 (but not by acetylcholine) was enhanced in vessels from LPS‐treated rats (see figure). The EC50 of Y‐27632 was reduced from 2.8 μM in control rings to 700 and 25 nM in LPS 6 and 24 h, respectively (p<0.05). The potentiation in Y‐27632‐induced relaxation was abolished by L‐NAME (100 μM), or 1400W (1 μM) at 6 but not 24 h after LPS. ODQ (10 μM) was effective at both 6 and 24 h. Western‐blot analyses revealed that protein levels of Rho‐A and ROCK‐II are increased in LPS‐treated animals.

Conclusions:

In spite of increased expression of Rho‐A and ROCK‐II, activation of this pathway is impaired in endotoxemic rats. This effect could be explained by increased production of nitric oxide and cGMP. We speculate that impaired Rho‐kinase pathway is involved in the hyporeactivity to vasoconstrictors observed in septic shock.
L‐type Ca2+ channels are expressed but not activated by KCl in rat small mesenteric veins
Authors: Thakali Keshari , Sonkusare Swapnil , Stimers Joseph R. , Rusch Nancy J.
Published: June 2021

Calcium channel blockers (CCBs) reduce afterload but not preload in vivo, suggesting that arteries and veins use different mechanisms to mediate contraction. We investigated this hypothesis using pressurized small mesenteric arteries (MA) and veins (MV) from male Sprague Dawley rats. Both MA and MV contracted robustly to 60 mmol/L KCl. Nifedipine (1 nmol/L – 1 μmol/L) and diltiazem (10 nmol/L – 10 μmol/L) each maximally reversed the KCl‐induced contraction in MA by 93%. Although we detected an abundance of L‐type Ca2+ channel mRNA and protein by RT‐PCR and Western blot, respectively, MV did not dilate significantly to either CCB. Whole‐cell patch‐clamp studies revealed that Ca2+ current (ICa) density in freshly dissociated smooth muscle cells from MV was 68% less than in cells from MA (MV, −3.8±0.7 pA/pF; MA, −11.7±3.0 pA/pF). However, the ICa was similarly sensitive to block by 100 nmol/L nifedipine (percent block: MA, 50%; MV, 60%). The KCl‐induced contraction in MA was prevented by removal of extracellular Ca2+, whereas 60±7% of the response persisted in MV. This persistent contraction was eliminated by EGTA (0.5 mmol/L) or thapsigargin (1 μmol/L). Thus, MV express functional L‐type Ca2+ channels, but these channels do not contribute to KCl‐induced contraction. Instead, a unique mechanism linking membrane depolarization to intracellular Ca2+ release may exist in veins.
Doxycycline improves vascular dysfunction and remodeling in renovascular hypertension via regulation of matrix metalloproteinase (MMP)‐2.
Authors: Castro Michele Mazzaron , Rizzi Élen , Lopes Lívia , Fernandes Karla , Bendhack Lusiane , Pitol Dimitrius , Gerlach Raquel , Tanus‐Santos José Eduardo
Published: June 2021

Altered activity of MMPs is implicated in the vascular remodeling of hypertension. We examined whether increased MMP‐2 plays a role in the vascular remodeling and dysfunction in the 2‐kidney, 1‐clip (2K‐1C) hypertension. Rats were treated with doxycycline (D) 30 mg/kg/day (or vehicle) by 8 weeks. Aortic rings were isolated to assess vascular reactivity. Quantitative morphometry of structural changes were studied in stained aortic sections. Aortic MMP‐2 levels and activity were determined by immunohistochemistry, gelatin/in situ zymography and antibody capture assay. MMP‐2/tissue inhibitor of metalloproteinases (TIMP)‐2 mRNA levels were determined by Real Time RT‐PCR. Doxycycline attenuated 2K‐1C hypertension (215 ± 8 vs. 167 ± 13 mmHg in 2K‐1C and D‐2K‐1C; P<0.01) and prevented the 35% reduction in endothelium‐dependent vasorelaxation found in 2K‐1C. Moreover, it also prevented the increases in media thickness, and was associated with lower media/lumen and cross sectional areas (all P<0.01). Increased MMP‐2 and Pro‐MMP‐2 levels and MMP‐2 activity were found in the aortas of 2K‐1C (p<0.05). A 21‐fold increase in the ratio of MMP‐2/TIMP‐2 mRNA expression was found in the 2K‐1C, whereas this ratio remained unaltered in D‐2K‐1C. Our results suggest that MMP‐2 plays a role in 2K‐1C hypertension and its structural and functional vascular changes, which were attenuated by doxycycline.