bluuebunny/crossref_metadata_2025 huggingface dataset:
Undaria pinnatifida extract improves postprandial glycemic control in Streptozotocin‐induced diabetic rats
Authors: Kim Youngsik , Nam Jeongsu , Lee Won Joon , Yoon Imsil , Yoon Joongsoo , Choi Hyunju
Published: June 2021

Undaria pinnatifida is a type of seaweed commonly consumed by Koreans and Japanese. This study investigated the effects of Undaria pinnatifida on the glucose tolerance of male adult Streptozotocin‐induced diabetic rats. The groups were divided into two: one group with a diet of 10% Undaria pinnatifida extract (UPEX), and the other as the control group without the extract. After a 4‐week feeding period, glycemic control was assessed by the intraperitoneal glucose tolerance test (ipGTT, 1 g glucose/Kg) and maltose oral loading test (2 g maltose/Kg). On the first day, s‐glucose levels during the entire period of ipGTT were similar in two groups. At week 1, however, postprandial glucose levels at 0, 30, 60 and 180 min after orally loading the maltose solution were lower in the UPEX group than those in the control group(p<0.05). At week 3, postprandial glucose levels at 0, 30, 60, 90, 120 and 180 min were also lower in the UPEX group than those in the control group (p<0.05). On week 4, the s‐glucose levels at 0, 30, 60, 90, 120 and 180 min during the ipGTT were significantly lower in the UPEX group compared to those corresponding values in the control group. These results suggest that Undaria pinnatifida extract may have potential effects on tolerating glucose challenge, and lower blood glucose increment after taking a meal, therefore, improve the postprandial glycemic control. (KRF Grant #F00014)
Effect of coffee intake on body weight and GSH/GSSG ratio in rats
Authors: Cho Youn‐Ok , Choi Eun‐Young , Jang Jin‐Young , Park Seo‐Young
Published: June 2021

To investigate the effect of coffee intake on body weight change and the ratio of GSH/GSSG, ninety six rats were fed either a control diet (C) or control diet with coffee water(12g/L) (CF) for 4 weeks. At the respective time points, animals were sacrificed by the decapitation under light anesthesia. Immediately following decapitation, blood was collected in heparinized tubes and centrifuged to separate the plasma. Liver, kidney, heart, spleen, abdominal fat and skeletal muscle were rapidly removed and weighed. The plasma and tissues were stored at ‐40 ¡É until analyzed. The significance of difference between C group and CF was tested using unpaired t‐test. Coffee intake was associated with a decrease in final body weight, feed efficiency ratio, hematocrit value, hemoglobin concentration, and the weight of abdominal fat, heart, spleen (p<0.01) and liver (p<0.05). The ratio of GSH/GSSG was not significantly different between C and CF animals. Thus, it is suggested that coffee intake may lead a weight loss with a decrease in abdominal fat but may not affect the antioxidant defense system in GSH/GSSG. Also, coffee intake may aggravate the hematologic profile and lead an anemia.
Chemical properties of color potato, potato and their starches
Authors: Hwa Kang Myung , Park Soo‐Jin , Lee Jea‐Soon , Moon Eun‐Young , Lim Hak‐Tae , Choi Mi‐Kyeong
Published: June 2021

In order to find various uses of color potato (Potato Valley, Co., Ltd., Bora Valley), the present study measured the general elements of the potato, the swelling power of starch made out of color potato, water holding capacity, DSC, SEM, X‐ray, and amylase and amylopectin contents, and also measured chromaticity using potato powder prepared through freeze drying. The water content was higher in color potato (87.7%) than in white potato (80.1%), and the crude protein content was around 3% higher in white potato (12.5%) than in color potato (9.2%). The vitamin C content was higher in color potato (18.8mg/100g) than in white potato (10.5mg/100g). The starch yield was 7.9% in color potato and 17.5% in white potato, and the amylase and amylopectin contents and water holding capacity were not much different between color potato and white potato. The results of starch X‐ray were Type B for both, and when starch structure was examined through SEM no significant difference was observed. In addition, the violet color of the color potato was anthocyan pigment and its content was 139.0mg/100g.
Vitamin D improves progesterone‐mediated reductions in edema following traumatic brain injury
Authors: Figueiroa Silvia M , Rich Nicholas J , VanLandingham Jacob W , Schepkin Victor D , Levenson Cathy W
Published: June 2021

Edema and intracranial pressure associated with traumatic brain injury (TBI) result in poor long‐term clinical outcomes. Thus, we need to improve our understanding of the mechanisms that result in edema and develop new treatments to target water accumulation. Progesterone (PROG) has been shown to reduce edema after TBI. Because 1, 25 dihydroxyvitamin D (D) and PROG both interact with nuclear RXR receptors to regulate gene expression, we hypothesized that acute treatments with the active form of vitamin D could be a useful adjunct to progesterone treatment. TBI was introduced by weight‐drop (300g) creating a 2mm medial frontal cortex contusion following craniotomy in male Fisher rats. Animals were then treated at 1 and 6 hrs post‐injury with vehicle, PROG (16mg/kg), D (2.5 µg/kg), or PROG+D. At 24 h, direct measurements of water accumulation around the site of injury showed that while D alone was ineffective, there was a 4% reduction in water accumulation in PROG treated rats, and a 9% reduction in PROG+D animals. Edema results from vasogenic water accumulation after blood‐brain‐barrier damage and cytotoxic water from cell damage. Water regulating protein aquaporin‐4 was not changed after treatment. Novel in vivo diffusion magnetic resonance imaging at 21 Tesla (21T) allowed us to visualize and distinguish vasogenic and cytotoxic edema. MRI results support that PROG +D protects against vasogenic and cytotoxic edema.
Manganese exposure and iron deficiency result in home cage behavior alterations in the developing rat
Authors: Anderson Joel , Erikson Keith
Published: June 2021

Both manganese (Mn) exposure and iron deficiency (ID) have been linked to alterations in behavior, such as anxiety and locomotion, due to neurochemical disturbances. To date, no study has looked at the longitudinal behavioral alterations in the unaltered home cage of animals exposed to Mn or ID. Therefore, we sought to examine the effects of ID and Mn exposure over a three‐week period to determine time and dose‐dependent behavioral changes. Twenty one day old male rats were randomly placed into one of four dietary treatment groups: control, iron‐deficient, control with Mn exposure (via the drinking water), and iron‐deficient with Mn exposure. Rats were monitored for 24 hours during weeks four, five, and six of the dietary protocol using the Clever Systems Home Cage Scan video surveillance system. Several novel findings emerged from this study. For example, although ID was associated with a decrease in total distance traveled over time as previously reported, Mn exposure was related to an increase in total distance traveled over time which is contrary to prior reports of reduced locomotor activity in Mn exposed animals. While studies have examined effects of either Mn exposure or ID over short observational periods, this is the first to examine these effects on exploratory, locomotor, and autonomic behaviors longitudinally over a 24‐hour period in the home cage environment.
Influence of supplemental quercetin and epigallocatechin 3‐gallate on immunity and inflammation
Authors: Maxwell Kendra Renee , Nieman David C. , Henson Dru A , Williams Ashley , McAnulty Steven , Jin Fuxia , Shanely Andrew , Lines Thomas
Published: June 2021

Quercetin is a flavonol with broad spectrum bioactive effects that include anti‐inflammatory, anti‐pathogenic, antioxidant activity, and immunoregulatory influences. We tested the influence of 1000 mg quercetin (Q) with or without 120 mg epigallocatechin 3‐gallate (EGCG), 400 mg isoquercetin, and 400 mg EPA‐DHA (Q‐EGCG) on changes in measures of immunity and inflammation before and after a 3‐day period of heavy exertion. 39 trained cyclists were randomized to placebo, Q, or Q‐EGCG, and ingested supplements in a double blinded fashion for 2 weeks prior to, during, and 1 week after a 3‐d period in which subjects cycled for 3 h/d at 57% Wattsmax. Blood and saliva samples were collected before and after 2 weeks supplementation, immediately following the exercise bout on the 3rd day and 14‐h post‐exercise. Two weeks supplementation resulted in a significant increase in granulocyte oxidative burst activity (GOBA) in Q‐EGCG relative to P and increases in plasma quercetin for Q and Q‐EGCG. Immediately after the 3rd exercise bout, significant decreases for C‐reactive protein, plasma IL‐6 and IL‐10 were measured in Q‐EGCG compared to P. Granulocyte colony stimulating factor (GCSF) and CRP were reduced in Q‐EGCG 14‐h post‐exercise. Two weeks supplementation with Q‐EGCG was effective in augmenting GOBA, and countering inflammation following 3 days of heavy exertion in trained cyclists. Grant support from Quercegen Pharma.
Effect of drink temperature and capsiate to the subjective sensation of body and ambient temperature during cold exposure
Authors: Okumura Yuka , Kondo Emi , Tai Shinji , Okamura Koji
Published: June 2021

Pre‐exercise warm‐up facilitates exercise performance by raising the muscle temperature. Since a declining muscle temperature results in a deterioration of exercise performance, warming up is thus especially important in a cold environment. This study investigated whether the drink temperature and capsiate, which is an ingredient of red pepper without a hot taste, ingested before warming up could improve the effect of warm‐up exercise. Five healthy males were exposed to 10oC ambient temperature with winter clothes for 30 min in a sitting position. Thereafter, the subjects ingested 200 mL of one of the following drinks; 10 oC flavored water (W10), 60oC flavored water (W60), 10oC with capsiate (C10) and 60oC with capsiate (C60) in random order with a crossover design. The subjective sensations of the body and the ambient temperature were measured with a visual analog scale for 30 min after ingestion. The sensation of body temperature after ingestion was higher after consuming drinks at 60oC than 10oC. There was no difference in the sensation to the temperature of drinks between W60 and C60, however the sensation of body temperature immediately after ingestion was significantly higher for C60 than W60 and tended to remain higher for C60 thereafter. The sensation to ambient temperature after ingestion was also higher for 60oC than 10oC and while there was no statistical significance it tended to be higher for C60 than W60 towards the end of the trial. These results suggest that the sensations of the body and the ambient temperature are higher after ingesting a hot drink than a cold drink and capsiate may effectively induce thermal sensations.
Differences in body composition and metabolism in stunted compared to normal height children in Jamaica.
Authors: Pilewski Kathryn , Soares‐Wynter Suzanne , Walker Susan , Hoffman Daniel
Published: June 2021

While the prevalence of undernutrition is decreasing worldwide, there is still a large number of undernourished children in many developing countries. Previous research has shown that stunted children are more likely to have impaired fat oxidation and greater fat mass following recovery and later in life. However, it is not clear what metabolic differences exist prior to recovery from undernutrition. Thus, we measured body composition (using anthropometry) and energy metabolism (using indirect calorimetry) in 102 stunted and normal height children living in Jamaica.

The children were split into stunted (< ‐2 HAZ) or control ( HAZ > ‐1 groups), both of whom had similar WHZ (‐0.68 ± 0.82 v ‐0.37 ± 0.76 kg). Differences between the groups were analyzed using Student's t‐Test and ANOVA. The stunted group had significantly lower fat mass (3.56 ± 0.89 v 4.58 ± 1.33 kg) and were older (7.86 ± 0.36 yrs v 6.83 ± 1.13 yrs) compared to the control group. REE was significantly lower in the stunted group compared to the control group (1118 ± 134 v 1323 ± 165 kcal/day), even when adjusted for lean body mass and sex (1168 ± 24 v 1298 ± 16 kcal/day). There were no significant differences in the respiratory quotient between the groups (0.83 ± 0.05 v 0.85 ± 0.05).

Based on the data presented, we conclude that stunted children have lower metabolic rates, independent of lean body mass, compared to normal height children of similar weight for height.
Partially hydrolyzed 100% whey infant formula and atopic dermatitis risk reduction: a systematic review of the literature
Authors: Alexander Dominik D , Schmitt Donald , Tran Nga , Barraj Leila , Cushing Colleen A
Published: June 2021

Numerous randomized trials and prospective intervention studies have reported a reduced incidence of atopic dermatitis (AD) among infants who were fed partially hydrolyzed 100% whey protein formula (PHF‐W) compared with infants who were fed intact cow's milk protein formula. Therefore, we conducted a systematic critical review of the literature to better understand this relation. A total of 17 clinical trials and intervention studies, representing 12 distinct study populations met our criteria for inclusion. Most of the studies incorporated study designs for which random assignment of infant formula was conducted. In virtually all studies, a reduced incidence of AD and/or atopic manifestations that included AD were observed. Throughout the first year of follow‐up, an approximate 50% risk reduction is observed. When data were pooled together from four studies of superior methodological quality that reported results specifically for AD, the cumulative incidence of AD was significantly lower (p < 0.05) among infants through at least three years of follow‐up in the PHF‐W group compared with infants in the intact cow's milk protein group. In healthy infants who are not exclusively breastfed, partially hydrolyzed 100% whey protein formula may be considered as a routine use formula alternative to help reduce the risk of atopic dermatitis. This work was partially funded by Nestle.
Body fat and fat‐free mass normalization by height
Authors: Rodríguez Patricia , Bermúdez Enrique , Macri Elisa , Ponce Graciela , Gamba Carlota , Juiz Natalia , Zeni Susana , Exeni Ramón , Friedman Silvia
Published: June 2021

Measurement of children's body composition has become increasingly common. However, attention has been focused on methods, and less on the way in which data is expressed.

Objective

to study the relationship between height (Ht) with fat mass (FM) and fat‐free mass (FFM), measured by DXA in preschool children; and calculate an independent index (FMI and FFMI) of Ht. Seventy five children (males= 31, females= 44) aged 3 to 5, from La Matanza, Province of Buenos Aires, Argentina, attending a day care center, were included. There were healthy according to anthropometric (WHO, 1999) and clinical evaluation. Ht data were collected and FM and FFM were measured using a Lunar DPX‐L, pediatric software. Pearson's correlation was used to examine the association between FM, FFM, FMI and FFMI with Ht, and linear regression to determine what power of height (Hty) results in a zero correlation between body composition and Ht.

Results

FM and FFM are associated with Ht (p=0.0222 and p< 0.0001, respectively) without differences between sexes. The slope of the regression line relating lgFM to lgHt was 1.49, while that for lgFFM to lgHt was 2.12. Thus, FM/Ht and FFM/Ht2 are uncorrelated with Ht (p=0.4063 and p=0.3695, respectively).

Conclusion

to compare children's body composition, normalization for body size is required. The indices FMI (FM/Ht) and FFMI (FFM/Ht2) would be appropriate for this purpose.

Awarded by UBACyT O‐008 and 015.
Dietary patterns of children aged 0‐59 months in Ukambani region of Kenya
Authors: Ndiku Mueni , Sabaté Joan , Singh Pramil , Jaceldo‐Siegl Karen
Published: June 2021

Objective

Severe drought struck Northern and Eastern Kenya in 2006 that forced ~10% of Kenyan population to face starvation, and subsequent malnutrition in children. We assessed the dietary patterns of children aged 0‐59 months in Mwingi (MW) and Makueni (MA), two drought‐stricken districts in Eastern Kenya. Mwingi district uses pearl millet as the main staple cereal while Makueni uses maize (corn).

Methods

We interviewed 403 mothers to collect dietary information about their children from 4 randomly selected locations in MW and MA. Surrogate 24‐hr recalls from 614 children (300 boys and 314 girls) were obtained, and subsequently entered using NDSR 2008. Nutrients were transformed to achieve normality. Intake of selected nutrients and food groups were compared between the two districts using t‐test.

Results

Average (mean ± SD) energy intake (1064+499 kcal) in MW was significantly higher compared to MA (966 + 470 kcal) (p<0.05). Intake of carbohydrate, protein, iron and most B vitamins were significantly higher in MW compared to MA; fat, animal protein and vitamin A were significantly higher in MA. On average, grains contribute 55‐60% of total energy intake in both districts. The contribution to total energy from grains, vegetables and milk were significantly higher in MW compared to MA.

Conclusion

Overall, children in Mwingi have higher nutritional intake compared to Makueni.
Serum phosphate predicts early mortality in HIV patients on ART in Zambia
Authors: Heimburger Douglas C , Nyirenda Christopher , Kabagambe Edmond K , Potter Dara , Bosire Claire , Zulu Isaac , Chisembele‐Taylor Angela
Published: June 2021

Objectives

Antiretroviral therapy (ART) programs in developing countries report high mortality rates in the first 90 days of ART. We hypothesize that acute hypophosphatemia, analogous to refeeding syndrome, occurs & is a risk factor for mortality.

Methods

We enrolled 148 adults with BMI <16 kg/m2 or CD4 <50/μL at ART initiation in Lusaka, Zambia. Serum phosphate, diet and anthropometrics were recorded 6 times in 12 weeks of ART.

Results

2.4% of participants had serum phosphate <0.65 mmol/L (<2 mg/dL) at baseline; this increased to 10.9% after 1 week but decreased again after another week. Mean phosphate dropped ‐0.16±0.33 mmol/L (p<0.001) between weeks 0 & 1. In median 86 days of follow‐up, 28 participants died (93/100 person‐yr). Higher baseline phosphate predicted improved survival (HR 0.15; 95% CI: 0.04‐0.56). This remained significant after adjustment for sex, BMI, & CD4 (HR 0.21; 0.06‐0.82). Sex (HR men vs. women 2.54; 1.05‐6.16) & BMI (HR/unit ↑ in BMI 0.74; 0.59‐0.93) also significantly predicted early mortality, but CD4 count did not. Baseline serum phosphate correlated with CD4 & intakes of protein & fat but not with sex, BMI, appetite, or intakes of energy or carbohydrate.

Conclusions

Hypophosphatemia occurs in persons with HIV/AIDS in Zambia and may worsen early in ART. Higher phosphate concentrations at ART initiation strongly predict improved survival during ART. Support: UAB CFAR, CIDRZ, NIH AI 076430.
Dietary intake, growth, and other characteristics in children with autism
Authors: Ekvall Shirley , Fugazzi Patricia , Stevens Florence
Published: June 2021

To determine which autistic characteristics may relate to nutritional intake, all of the children with autism for that year, 87 children, aged 1 to 13 years, were evaluated. Ages were 26% 16 months‐3 years, 58% 3‐5 years and 16% 5‐13 years. Anthropometric measures were obtained according to standardized procedures. Population ethnicity was 60% Caucasian, 33% African American, and 7% Other. Eighty % were male and 20% female. Mothers were 21‐30 years. Results showed: picky eating 70%, allergies 5%, fruit/vegetables low in 74%, whole grains low in 68%, calories were adequate, anthropometric measures followed the growth curve for age. Nutritional assessment/ counseling is needed for these highrisk children.
Mechanisms of iron release from lysosomes
Authors: Nguyen Annie , Zhao Ningning , Morrison Chevaun , Gonzalez Angelica , Sauble Eric , La Alice , Linder Maria C. , Knutson Mitchell
Published: June 2021

Stored cellular Fe is made available and recycled at least partly through lysosomal degradation of cytoplasmic ferritin (Ft) after autophagy, but how Fe returns to the cytoplasm is unknown. Divalent metal transporter 1 (DMT1) is associated not just with transferrin‐related endosomes but also with lysosomes, and the latter is true for Zip 8, which might also be an Fe transporter. To begin the determine whether one or both DMT1 and Zip8 might be involved, we first established that Zip 8 transfection into HEK cells (with low endogenous levels) enhanced Fe uptake, determined with 59Fe‐citrate. We then knocked down expression of Zip 8 with siRNA in rat hepatoma cells, and studied the accumulation of 59Fe in lysosomes separated from cytoplasmic Ft on iodixanol gradients (Kidane et al, Am J Physiol 291: C445, 2006). To track and move Ft into lysosomes, we pretreated cells with 59Fe‐ferric ammonium citrate for 24h then induced Fe depletion with deferoxamine (as previously). Compared with scrambled siRNA, 80‐90% knockdown of Zip 8 mRNA was associated with about 35% greater retention of 59Fe in lysosomes. Using confocal microscopy on HepG2 cells, we also determined that Fe depletion rapidly increased colocalization of DMT1 with the lysosomal marker (LAMP2). We conclude that both of these transporters could be involved in the return of lysosomal Fe to the cytoplasm. Supported in part by PHS Grants RO1 HD 46949 (MCL) and RO1DK080706 (MK).
Soy protein influences plasma lipid profiles but does not reduce interleukin‐6 inflammation marker in dextran sodium sulfate treated rats
Authors: Marsh Tanya G , Straub Rachel , Villalobos Fatima , Hong Mee Young
Published: June 2021

Clinical and experimental data have indicated that soybeans, soybean constituents and soy isoflavones have health implications in preventing cardiovascular disease (CVD). Inflammation and serum lipid profiles play pivotal roles in the development of CVD. We investigated the role of soy protein on lipid profiles, and inflammation indicated by the interleukin‐6 (IL‐6) level in 3% dextran sodium sulfate (DSS)‐treated rats. The soy protein isolate used in the study contained 825 μg, 485 μg and 181 μg of genistein, daidzein and glycitein per gram of soy protein isolate, respectively. Soy protein decreased total cholesterol (P<0.001) and LDL‐cholesterol (P<0.001) compared to casein. DSS treatment increased serum IL‐6 levels (P<0.05) but IL‐6 levels were not significantly different between the diet groups. Interestingly, soy protein‐fed rats had lower final body weights (P<0.05), along with lower liver (P<0.01) and epididymal fat weights (P<0.05) despite no differences in food intakes. These results demonstrate that soy protein isolate may lower the risk of CVD by improving serum lipid levels and lowering body weights. Further study is warranted to examine the mechanism of soy protein in body weight reduction and/or alteration of body composition. This study was supported in the Advanced Nutrition Lab at San Diego State University and funded by SDSU UGP and CHNR08‐810.
Molecular Diagnostics for Identifying an Increased Likelihood of Recurrence of Breast Cancer
Authors: Wittliff James L. , Andres Sarah A. , Kerr D. Alan
Published: June 2021

Existing prognostic factors based on histological and clinical parameters result in under‐treating some patients at high risk of recurrence and over‐treating others that may not benefit from chemotherapies. Current gene expression tests are based on gross tissue biopsies exhibiting cellular heterogeneity. Our goal was to develop a subset of genes (5‐15) based on UniGene (NCBI) analysis and published data using whole tissue, while exploiting larger gene sets derived from cancer cells collected by LCM (e.g., Ma et al., Breast Cancer Res Treat 82:S15, 2003). Applying the UniGene database, comparisons of molecular signatures reporting >2500 sequences were reduced to 5‐15 gene subsets of candidate cancer and stromal cell genes. Each gene was validated by qPCR and correlated with clinical outcome. A molecular signature from cancer and stromal cell candidate genes predicted early breast cancer recurrence. A separate 13 gene signature in ER+ breast cancer correlated with disease‐free and overall survival. Gene candidates were analyzed on the Ziplex® Automated Workstation (Xceed Molecular) with a TipChip array. Our results suggest a molecular diagnostic test based on expression of gene subsets predicts clinical outcome. Supported in part by grants from Phi Beta Psi Charity Trust, University of Louisville's Office of Technology Transfer and Xceed Molecular Corp. SAA & DAK are recipients of IPIBS Graduate Fellowships.
Exposure of swine to nonpenetrating blast results in reactive astrogliosis, fiber degeneration, and upregulation of inflammatory markers
Authors: Tong Lawrence , Januszkiewicz Adolph J , Swauger Peter V , de Lanerolle Nihal C , Kim Jung H , Gyorgy Andrea B , Agoston Denes V , Bandak Faris A , Ling Geoffrey S , Parks Steven A
Published: June 2021

Blast‐induced nonpenetrating brain injury (bTBI) has become increasingly prevalent in recent years. The arsenal of available treatments continues to be inadequate while the mechanism of injury remains equivocal. Although similar studies have been reported on small animal models using fluid percussion and weight‐driven devices, none have been conducted on large animals under real‐world conditions. We exposed swine to varying levels of focused explosions within a shock tube in order to quantify the intensities needed to produce minimum detectable and maximum survivable bTBI. In addition, improvised explosive devices were detonated near swine placed within a surrogate Humvee, in a simulated building/street setting, or in an open‐field scenario. All animals were exposed left‐laterally to incident explosion. Early histological findings are distinctly different from other causes of TBI and include activation of astrocytes in the hippocampus and overlapping fiber tracts as well as fiber degeneration in the thalamus and anterior cerebellar vermis, that do not clearly correlate with exposure lateralization. Preliminary proteomic analyses of cerebrospinal fluid, blood and brain tissue indicate blood‐brain barrier disruption along with a prominent inflammatory response.
Puma is a critical regulator of acute ethanol‐induced neuroapoptosis
Authors: Ghosh Arindam , Klocke Barbara J , Roth Kevin Aaron
Published: June 2021

Intrauterine exposure to ethanol causes a dysmorphogenic neuropathological syndrome in humans. The toxic effects are most pronounced during the synaptogenesis period of nervous system development that predominantly occurs prenatally in humans but postnatally in rodents. To define the critical upstream pro‐apoptotic mediators of acute ethanol‐induced neuroapoptosis, we examined the effects of ethanol administration in wild type mice and mice with targeted gene disruptions in puma, bax, noxa and p53.Apoptosis as evidenced by a robust increase in cleaved caspase‐3 like immunoreactivity was observed at 6 hours post treatment. Bax deficient mice however did not show an increase in cleaved caspase‐3 levels indicating that Bax deficiency is protective in this process. Puma knockout mice also did not show an elevation in cleaved caspase‐3 levels after treatment with ethanol, though p53 deficient animals treated with ethanol showed comparable caspase‐3 levels as p53 heterozygous mice or wild type mice.

These results indicate that acute ethanol‐induced neurodegeneration is mediated by Puma and Bax and is independent of regulation by p53.The transcription factor that mediates this process remains to be determined.

This work was supported by UAB Neuroscience Core facilities (NS 47466 and NS57098) and NIH grant NS35107 (KAR)
Intravital imaging in a transgenic mouse model of breast cancer
Authors: Resau James H , Williams Bart , Lindvall Charlotta , Zylstra Cassandra , Looyenga Brendan , VandenBeldt Kristin , West Rich
Published: June 2021

Currently there are few ways to complete intravital imaging of well characterized transgenic mouse models. We are developing an assay to monitor by fluorescence, a transgenic, murine, mammary carcinogenesis model of breast cancer that expresses the well characterized beta‐galactosidase marker. We are utilizing a method to observe the marker in cells, and will apply the method to developing lesions and histology preparations without terminating the biologic development of the tumor and hopefully treatment induced regression in order to determine efficacy.

We have crossed MMTV WNT 1 mice with mice expressing the β‐galactosidase transgene under the control of a β‐catenin‐responsive promoter (Bat‐Gal). These animals have a well characterized, developmental pattern that originates in the mammary fat pad. The β‐catenin protein is stabilized in MMTV‐Wnt1‐induced mammary tumors. Building on Tung et al (2004), we quantified Wnt1‐induced mammary tumors by measuring the activity of β‐gal with DDAOG, a red fluorescent signal. Our model uses instruments (cri‐inc.com) designed for multispectral, in‐vivo fluorescence imaging. We un‐mix the image cubes and separate the DDAO and background components. We can quantify expression by using software programs (Kort et al, 2003). Upon sacrifice, the animals will be further characterized using spectral and confocal microscopes to develop 3D reconstructions of the tumors.
AAV‐mediated vascular smooth muscle‐specific knockdown of Cav1.2 as a long‐term anti‐hypertensive therapy
Authors: Pang Li , Sonkusare Swapnil , Thakali Keshari , Melchert Russell , Roberto Kerrey , Palade Philip
Published: June 2021

Hypertension is a chronic disease, but currently all antihypertensive drugs have relatively short half‐lives. To improve patient compliance, we hypothesize that AAV‐mediated Cav1.2 siRNA gene therapy which selectively reduces the abnormal expression of Cav1.2 in vascular smooth muscle cells (VSMCs) will provide a long‐term antihypertensive effect. We previously reported a miRNA‐based shRNA expression cassette driven by a VSMC‐specific enhancer/promoter significantly reduced Cav1.2 expression and function in cultured VSMCs but not in non‐VSMCs tested. In this study, we subcloned the VSMC‐specific siRNA expression cassettes into an AAV vector and injected the viruses into angiotensin II‐induced hypertensive mice by tail vein injection. Compared to control virus‐treated mice, blood pressure of AAV/Cav1.2 siRNA‐treated mice decreased ~30 mm Hg and remained reduced for = 4 weeks. L‐type Ca currents from isolated mesenteric artery VSMCs significantly decreased in AAV/Cav1.2 siRNA‐treated mice (‐4.58±1.51 pA/pF vs. ‐10.96±1.89 pA/pF in control virus‐treated mice, p<0.05). Expression of Cav1.2 siRNA was VSMC‐selective since GFP (an expression marker, driven by the same promoter as the Cav1.2 siRNA) mRNA was only detected in arteries, but not in heart, brain, or liver.

Conclusion

AAV‐mediated VSMC‐specific knockdown of Cav1.2 may provide a long‐term therapy for hypertension. Supported by NIH HL63903.
The restoration of cerebrovascular myogenic function after hemorrhagic stroke development in stroke prone hypertensive rats
Authors: Smeda John Stanislaw , Daneshtalab Noriko
Published: June 2021

Hemorrhagic stroke (HS) development in Kyoto Wistar stroke prone hypertensive rats (SHRsp) is associated with the loss of pressure dependent constriction (PDC) in middle cerebral arteries (MCAs). We attempted to restore PDC by administering oral Captopril (CAP, 50 mg/kg/day) or Losartan (LOS, 35 mg/kg/day) treatment after stroke. MCAs were studied using a pressure myograph. MCAs from SHRsp with HS did not elicit constriction to a 100 mmHg pressure step or in response to PKC activation (phorbol dibutyrate 1μM, with 3μM nifedipine). Elevations in [K+]o (80mM) produced depolarization with attenuated constriction. The MCAs from post stroke SHRsp also could not utilize the sarcoplasmic release of Ca2+ to produce constraction (vasopressin 0.12 μM, with 3μM nifedipine). Post stroke CAP or LOS treatment restored these functions to pre‐stroke levels within 7 days without altering blood pressure. These functions deteriorated after 30 days of CAP treatment but were maintained past 100 days of LOS treatment. We believe that the loss of PDC after HS is produced by a dysfunctional PKC system and the presence of voltage gated Ca2+ channel that exhibits an attenuated response to depolarization. The enhanced ability of LOS to restore these functions after HS may be related to the physiological benefits of maintaining high plasma angiotensin II in combination with AT‐1 receptor blockade.
Impaired function of prejunctional adenosine A1 receptors in sympathetic nerves supplying mesenteric arteries in deoxycorticosterone acetate (DOCA)‐salt hypertensive rats
Authors: Sangsiri Sutheera , Dong Hua , Xu Hui , Galligan James J
Published: June 2021

Increased sympathetic nervous system activity contributes to DOCA‐salt hypertension in rats. ATP and norepinephrine (NE) are released from sympathetic nerves. NE acts at prejunctional α2 adrenoceptors to inhibit NE release and prejunctional α2 adrenoceptor function is impaired in DOCA‐salt rats. Adenosine is a product of ATP hydrolysis and adenosine acts at prejunctional A1 adenosine receptors to inhibit NE release. We tested the hypothesis that prejunctional A1 receptor function is impaired in sympathetic nerves associated with mesenteric arteries (MA) and veins (MV) of DOCA‐salt rats. Electrically‐evoked NE release and constrictions of MA and MV were measured in vitro using amperometry and video microscopy, respectively. Adenosine and N6‐cyclopentyl‐adenosine (CPA, A1 receptor agonist) constricted MV but not MA. Adenosine and CPA caused concentration‐dependent (0.001‐10 μM) inhibition of MA constrictions. Adenosine and CPA concentration responses curves (CRCs) in DOCA‐salt MA were right shifted (P<0.05) compared to control MA. CPA inhibited NE release in MA and MV. The CPA CRC in DOCA‐salt MA, but not MV, was right shifted compared to control MA. These data indicate that there are prejunctional A1 receptors in MA and pre and postjunctional A1 receptors in MV. Prejunctional A1 receptor function is impaired in MA, but not MV, from DOCA‐salt rats. These data indicate that prejunctional α2 adrenoceptors are not selectively affected in DOCA‐salt hypertension. There is a more general disruption of prejunctional mechanisms controlling sympathetic neurotransmitter release.
GHB‐ and baclofen‐induced hypothermia in mice: interactions with the GABA‐B receptor positive modulator CGP7930, the GABA‐B receptor antagonist CGP35348, and the NOS inhibitor L‐NAME
Authors: Koek Wouter , Campos Peter S. , France Charles P. , Cheng Kejun , Rice Kenner C.
Published: June 2021

CGP7930, a GABA‐B receptor positive modulator, enhances GABA‐B receptor agonist‐induced loss of righting in mice (Carai et al., 2004). The present study examined the generality of CGP7930's ability to enhance in vivo effects of GABA‐B receptor agonists.

Baclofen and GHB both produced hypothermia (ED50: 4.8 and 270 mg/kg, resp.). When given alone, CGP7930 also produced hypothermia, but was less potent (ED50: 100 mg/kg) and less effective than baclofen. CGP7930‐induced hypothermia was not attenuated by the GABA‐B receptor antagonist CGP35348, which blocked baclofen‐induced hypothermia. CGP7930 did not enhance the hypothermic effects of baclofen and GHB. In contrast, the NOS inhibitor L‐NAME had no effect on body temperature when given alone but enhanced baclofen‐ and GHB‐induced hypothermia.

The finding that CGP7930 produced hypothermia when given alone, and that this effect was not attenuated by CGP35348, suggest that, in vivo, CGP7930 may behave as an allosteric agonist at GABA‐B receptors. However, the results do not provide evidence that CGP7930 also behaves as a positive modulator at GABA‐B receptors mediating baclofen‐ and GHB‐induced hypothermia. Together with previous findings, the present results confirm the utility of CGP7930 for examining the role of GABA‐B receptor modulation in the in vivo effects of GABA‐B receptor agonists.

Supported by DA15692 (WK), DA17918 (CF)
Alterations in D2 dopamine receptor internalization in the presence of the Na+/K+‐ATPase
Authors: Hazelwood Lisa A , Free R Benjamin , Cabrera David M , Sibley David R
Published: June 2021

Using a co‐immunoprecipitation assay for dopamine receptors (DARs) coupled with mass spectrometry (MS) sequencing, we previously identified the Na+/K+‐ATPase (Na+ pump, NKA) as a member of both the D1 and D2 DAR signalplexes. Using fluorescent‐labeled D2 DAR and NKAα1 constructs in confocal microscopy experiments, co‐localization of the two proteins was observed at the plasma membrane. Dopamine stimulation of RFP‐tagged D2 DAR‐expressing cells resulted in marked DAR internalization twenty minutes after drug treatment. Interestingly, when GFP‐tagged NKAα1 was co‐expressed, the D2 DAR internalization twenty minutes after dopamine treatment was diminished. To achieve a more quantitative assessment of receptor expression, cell‐surface binding assays were performed in the presence and absence of over‐expressed NKA. These assays revealed a 45% decrease in cell‐surface D2 DAR number twenty minutes after dopamine treatment, and a 30% decrease in receptor number when the NKA was also present. Longer dopamine incubations enhanced D2 DAR internalization, but co‐expression of NKA prohibited complete internalization as compared to D2 DAR alone. Current studies are underway to determine the impact of the DAR‐NKA complex on receptor recycling, and the role of NKA in lipid rafts on receptor activation and internalization.

Support contributed by the NINDS intramural program.
Activation of protein kinase B/Akt and endothelial nitric oxide synthase mediates Lycopus lucidus‐induced endothelium‐dependent vascular relaxation
Authors: Lee Yun Jung , Lee So Min , Kang Dae Gill , Kim Jin Sook , Lee Ho Sub
Published: June 2021

We recently reported that aqueous extract of Lycopus lucidus Turcz. (ALT) suppressed ROS and NF‐κB activation, resulting in prevention of high glucose‐induced vascular inflammation in vitro. In the present study, low‐density lipoprotein receptor deficient (LDLr‐KO) mice were fed a high fat diet (HFD) consisting of 60 kcal% fat, with or without ALT 200 mg/day/kg, for 14 weeks. HFD‐LDLr‐KO mice were treated with ALT were completely normalized by lowering glucose but were unaffected by body weight. ALT reduced atherosclerosis in HFD‐LDLr‐KO mice without affecting plasma lipid levels. ALT treatment caused endothelium‐ and nitric oxide (NO)‐dependent relaxation of aortic rings in HFD‐LDLr‐KO mice. Pretreatment with L‐NAME, a non‐specific NOS inhibitor, attenuated the protective action of ALT on high glucose‐induced adhesion molecule expression, suggesting a potential role of NO signaling. Further experiments demonstrated that high glucose‐induced adhesion molecules expression is inhibited by using Akt1 siRNA. ALT increased eNOS expression, while decreased vascular cell adhesion molecule‐1 and monocytes/macrophages marker expressions in HFD‐LDLr‐KO mice. In conclusion, chronic treatment of ALT improved endothelium‐dependent vascular relaxation as well as exhibited anti‐inflammatory effect in diabetic atherosclerotic mouse model, independent of effects on plasma lipids.
Sympathectomy does not cause supersensitivity to noradrenaline in rat prostate
Authors: Docherty Jim , Bexis Sotiria , Liu Cecil , Merza Jasem , Dang Michelle
Published: June 2021

Chemical sympathectomy classically produces a denervation supersensitivity assumed to be due to a general increase in receptor number on smooth muscle. However, we have suggested that denervation results rather in a spread of synaptic alpha1D‐adrenoceptors, resulting in an increased proportion of this subtype. We now examine whether these effects of sympathectomy are dependent on the prior presence of innervated alpha1D‐adrenoceptors by studying rat prostate in which innervated receptors are reported to be alpha1A. Animals were sympathectomised by 6‐hydroxydopamine injection. Contractions to nerve stimulation in prostate were markedly reduced by the alpha1A‐ antagonist RS100329, which was more potent than the alpha1D‐antagonist BMY 7378. Norepinephrine (NE) potency at producing contractions was unchanged by sympathectomy, so that denervation supersensitivity did not occur. RS100329 significantly shifted the potency of NE to a similar extent in tissues from vehicle‐treated and sympathectomised animals. BMY 7378 revealed a small alpha1D‐adrenoceptor component to NE contractions in sympathectomised but not control. Since supersensitivity does not occur in prostate, a tissue without a significant amount of alpha1D‐adrenoceptors, this may imply that the presence of functionally important innervated alpha1D‐adrenoceptors is required for denervation supersensitivity.
Selective colonic irradiation induces urinary bladder overactivity
Authors: Zabbarova Irina , Epperly Michael , Greenberger Joel , Kanai Anthony
Published: June 2021

Afferent cross‐sensitization is considered to be one of the major factors contributing to the pathologies involved in pelvic pain although the precise mechanism is unknown. We have developed a mouse model for studying afferent cross‐sensitization of the colon and the urinary bladder. This was accomplished by momentarily withdrawing and selectively irradiating a section of the descending colon. The irradiation beam (2 ‐ 10 Gy) was focused on the gut without affecting other organs. Using this model we have demonstrated that selective colonic irradiation induces bladder overactivity. At 1 to 2 weeks following irradiation, cystometric studies on decerebrated mice demonstrated non‐voiding contractions (3 to 6 per contractile interval) and decreased intercontraction intervals (from 12±3 min in controls to 1±0.5 min in irradiated mice).

We have previously reported on the protective effects of mitochondrially targeted or intravesical nitric oxide synthase (NOS) antagonists during bladder irradiation. NOS activation is hypothesized to be a primary event in irradiation induced damage. However, the mechanism of cross‐sensitization may involve antidromic release of transmitters from bladder afferents in that NOS antagonists were not protective when the gut was irradiated. Accordingly, ongoing studies are directed at characterizing the mechanism of bladder‐colon cross‐sensitization. Funded by NIH DK07108.
Functions of Transmembrane Domain 6 in Human Melanocortin‐4 Receptor
Authors: Fan Zhen‐Chuan , Tao Ya‐Xiong
Published: June 2021

The melanocortin‐4 receptor (MC4R) is a G protein‐coupled receptor critically involved in regulating energy homeostasis. Activation of the MC4R leads to decreased food intake and increased energy expenditure. Loss‐of‐function mutations in the MC4R gene were the most common form of monogenic obesity. Despite the important therapeutic potential of the MC4R in obesity treatment, the structure‐function relationship has not been adequately investigated. In this study, we investigated the functions of the transmembrane domain (TM) 6 of the MC4R using alanine‐scanning mutagenesis. A total of 31 mutants were generated. The ligand binding and signaling properties of these mutants were measured. Maximal binding capacities were decreased by 50% for P260A, F261A, H264A, L265A, Y268A, and S270A compared to the wild type MC4R. Maximal signaling after stimulation with the superpotent agonist NDP‐MSH was decreased for I266A and F267A. Six mutants located at the cytoplasmic half of TM6, including M241A, I245A, T246A, T248A, I249A, and L250A, have significantly increased constitutive activities, suggesting that these residues are important for constraining the wild type MC4R in inactive conformation. In summary, we identified six residues at the cytoplasmic end of TM6 that are important for constraining the WT MC4R in inactive conformation, and F267 as critical for agonist‐stimulated signaling.
Characterization of α1A‐adrenergic receptor splice variants in human aortic and pulmonary arterial smooth muscle cells
Authors: Aungst Timothy Dy , Bachman Adam , Prinster Steven C. , McCune Dan F.
Published: June 2021

Three α1‐adrenergic receptor (AR) subtypes have been cloned and characterized, the α1A‐, α1B‐, and α1D‐AR. Additionally, 16 splice variants of the α1A‐AR have been isolated in various human tissues. We characterized the expression and function of α1‐ARs in human aortic and pulmonary arterial smooth muscle cells. The expression α1‐AR mRNA was assessed with RT‐PCR utilizing subtype and splice variant specific primers. Response to agonist was assayed by measuring increases in intracellular calcium ion concentrations in cells loaded with Fura‐4. Both aortic and pulmonary cells express mRNA for the α1A‐, α1B‐, and α1D‐ARs. Interestingly, message for α1A‐AR splice variants 1‐3, but not 4, was detected in both cell types. Both cell types displayed increases in intracellular calcium in response to treatment with the α1‐AR selective agonist phenylephrine, suggesting functional receptor expression. Simultaneous expression of all three α1‐AR subtypes is consistent with previous reports, while the significance of α1A‐AR splice variant expression is currently unknown. Further study of these cell types with a variety of agonists and antagonists to explore multiple signal transduction pathways is required.

Supported by a Type I grant, Wilkes University
Comparison of the effects of agonists and antagonists at peroxissome proliferator‐activated receptor gamma and angiotensin II receptors on murine preadipocytes and cardiac sympathetic neurons
Authors: Pinheiro Helder , Bastos Ricardo , Silva Filipa , Calhau Conceição , Moura Daniel
Published: June 2021

The angiotensin II (AngII) receptor antagonist telmisartan, activates the peroxisome proliferator‐activated receptor gamma (PPARγ) which is a binding site for the antidiabetic thiazolidinedione. The aim of this study was to characterize the action of different AT1 receptor antagonists on PPARγ‐mediated effects and, reversely, to characterize the action of the PPARγ agonist, rosiglitazone (Rz), on AngII receptor‐mediated effects. Adipogenesis (Oil red O staining) in a preadipocyte cell line (3T3‐L1) was used to test the actions of several AT1 antagonists on PPARγ. Facilitation of tritiated noradrenaline (3H‐NA) release induced by rat sympathetic nerve stimulation was studied to test the actions of Rz on AngII receptors. Rz (300 nM) but also ZD7155 (1 μM) and candesartan (10 μM) promoted adipogenesis which was blocked by the PPARγ antagonist GW9662 (3 μM). Neither losartan nor eprosartan had any effect on adipogenesis. Rz (30‐300 nM) caused a concentration‐dependent facilitation on the evoked overflow of 3H‐NA. These results suggest that ZD7155 and candesartan promote PPARγ‐mediated adipogenesis, while losartan and eprosartan are devoided of this action. Conversely, the facilitatory action of Rz on 3H‐NA release from sympathetic nerve terminals suggests that Rz and AngII may share binding sites or signal transduction pathways.

Supported by "Investigação na Pré‐graduação, Universidade do Porto‐CGD"