bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of a peer‐led nutrition education intervention among college students
Authors: Bradley Kristen , Silliman Kathryn , Morris Michelle Neyman
Published: June 2021

A peer‐led nutrition education intervention, Leaders in Nutrition Knowledge and Support (LINKS), was evaluated for its effectiveness among college students whose eating habits are typically inconsistent with current dietary recommendations. The LINKS curriculum was delivered by a nutrition graduate student and ten undergraduates to various student groups and included six sessions on basic nutrition, fueling the student athlete, disordered eating, healthy eating when dining out, supermarket tour and cooking demonstration. 252 subjects, 201 females (80%) and 51 males (20%) participated. On average, across all six sessions, subjects responded favorably to the ten survey statements (Likert scale 1 = strongly disagree to 4 = strongly agree) related to program effectiveness. Mean scores ranged from 3.77 ± .57 (content was appropriate for college population) to 3.2 ± .87 (likely to use distributed handouts in the future). The cooking demonstration received the highest overall effectiveness (3.81 ± .2) and was higher (p<.001) than the basic nutrition session (3.37 ± .61). Overall effectiveness of the remaining sessions ranged from 3.56–3.65. LINKS was well received and can be replicated on other college campuses. In particular, cooking demonstrations effectively provided information and developed skills related to planning and preparing healthy meals. Funding provided by a Lantis University Professorship.
Love Your Body, Love Yourself: Development of a nutrition and physical activity curriculum for a school‐based obesity intervention in high‐risk girls ages 11–17 years.
Authors: Uhley Virginia Ellen , Lasichak Andrea , Malone Jennifer , LaVerne Catalina , Harris Mary Elizabeth , Kato Kumiko , Grinnell Laura , Cooper Erica , Schaad Dana , Nwankwo Chinazo , Abatemarco Andrea , Beddingfield Stefanie
Published: June 2021

Overweight adolescents have a 70% chance of becoming overweight or obese adults. One of the consequences of overweight as perceived by the children themselves is social discrimination. This is associated with poor self‐esteem and depression. The Love Your Body, Love yourself nutrition and physical activity weight loss curriculum was designed for girls ages 11–17 years who were involved in a residential treatment center for mental health, substance abuse, trauma recovery, and other psychosocial issues. Within this residential program, 50% had a BMI classification of obese, 30% had high blood cholesterol levels, 20% had hypertension, and 40% had asthma. The residential population was 30% Caucasian, 64% African American and 2% Hispanic girls. The overall theme of the curriculum development was focused on the importance of different body parts as they relate to overall health, taking away from the attention on outward appearance. The lesson plans are divided into weekly modules, each module includes a specific body part and how nutrition and physical activity play a role with the overall health of their bodies. Nutrition and physical activity lesson content was also designed based on results from survey data that was collected from the girls regarding their nutrition and physical activity knowledge and level of interest. These modules are designed to be integrated into an 8 month school‐based wellness program.
Weight loss and dietary improvements following the Healthy Lifestyle Change Program, a community‐based intervention for adults with developmental disabilities
Authors: Zeldin Ari Stefan , Shihady Ida R , Garro Nicole , Lehrer Danise , Eis Jean , Bazzano Lydia A , Bazzano Alicia T
Published: June 2021

Objective:

People with developmental disabilities (DD) are at high risk for obesity due to inactivity, poor diet and medication effects. Our goal was to evaluate the effect of an intervention designed for adults with DD on weight loss and dietary habits.

Methods:

A pre‐post test design was used to evaluate the Healthy Lifestyle Change Program (HLCP), a 7‐mo program to improve dietary habits and reduce obesity in adults with DD with BMI≥25. Forty‐five participants completed nutrition and exercise sessions twice weekly for 7 mos. Abdominal girth, dietary habits, nutrition knowledge, weight satisfaction, weight and BMI were measured at baseline and 7 mos.

Results:

Two‐thirds of participants maintained or lost weight. Mean weight loss was 3 lbs (p=0.03) and BMI change was −0.5 (p=0.04). Abdominal girth decreased by a mean of 1 in. (p=0.005). An increase in weight satisfaction was reported by 43% (p=0.0001). Participants reported significantly improved diet. Water and nutrient‐dense food intake increased significantly (p<0.0001), while intake of energy‐dense snacks, fast foods and soda decreased. Nutrition knowledge improved on recommended servings of fruits, carbohydrates, whole grain products and risks of fried foods (p≤0.05)

Conclusion:

The HLCP is an effective strategy to improve dietary habits and knowledge and decrease abdominal girth and weight for adults with DD.

Source of Support: CA Dept of Dev S vcs
Nutrition knowledge and survey among urban college students
Authors: Tan Irene , Ho Charlene C , Ho Yen Kuang
Published: June 2021

Written expression of nutrition knowledge may be an informative indicator to assess the nutrition practices among urban college students. The objective of the “Self‐Assessment” questionnaire survey instrumentation (Adapted Nutrition Action Health Letter, CSPI) is to identify the nutrition habits of urban college students. The questionnaire was distributed among San Francisco City College students enrolled in the Fall 2007, Human Biology class. The survey does not attempt to include all aspects of the student's diet. In addition, the survey does not measure precisely the key nutrients. Each answer rates the students’ nutritional eating habits from a scale +4 (best) to –4 (worst). The average of the total score for 39 questions for male and female are 2.83 and 16.49, respectively. There was no statistical difference in Group 1 between male and female, but the remainder Groups (2–4) show statistical difference (p < 0.05) between both genders. Male had 52%, 31%, 17% and female had 24%, 43%, 31% for Group 2, 3, 4, respectively. BMI was calculated to show no significant different in BMI between male (23.32) and female (22.98), but the pattern of distribution remains normal and show variations between male and female students. The percentage of students with BMI<18.5 between female and male are statistically different, 12% and 2%, respectively. Both male and female students with a BMI > 30 are 6.5%, lower then U.S. general population. Waist circumference measurements were collected, 87 cm (male) 73 cm (female). The survey instrument found college students know how to improve nutrition and physical activity considerably, but there is lots of room to improve.
The effect of Zn supplementation on Zn status depends on the iron status in physically active women
Authors: Lee Okhee , Kim Mijin , Moon Jongwha
Published: June 2021

Exercise can cause change of mineral levels by electrolyte and water loss through sweat. Zinc is known to be involved in exercise‐induced metabolic responses during exercise, so adequate zinc nutrition seems to improve exercise tolerance. The prevalence of iron deficiencies are known to be high in some physically active women. The iron nutrition status influences Zn status through competing for a common transporter in the intestinal absorption. This study investigates whether the iron status can influence the effect of Zn supplementation on serum Zn level. Female basketball collegiate players were supplemented 2 months with zinc 50mg as zinc gluconate. Bloods were collected before and after exercise at the beginning and end of 2‐mo. study period. The serum concentrations of Fe and Zn were measured by neutron activation analysis method. Half of athletic females before Zn supplementation showed a state of iron depletion (ID, serum ferritin <20 μg/L). Before supplementation, the serum Zn concentration was lower in athletes with ID than athletes without ID. After 2‐mo. Zn supplementation, the serum Zn level was higher in athletes with ID than without ID. showing lower Zn status with normal Fe status. Conclusively, this study suggests that the effect of Zn supplementation on serum zinc level in physical active women can depend on the iron status before supplementation. The 2‐mo. Zn supplementation can enhance the serum Zn concentration of athletic females with low iron status, but not with normal iron status. This study was supported by the MOST of Korea
Cytomorphology of myxopapillary ependymoma
Authors: Takei Hidehiro , Powell Suzanne Z.
Published: June 2021

Myxopapillary ependymoma (MPE) is a rare distinctive variant of ependymoma (EP) occurring predominantly in the conus‐cauda‐filum terminale region of adults. Cytologic features of MPE have rarely been described. Touch imprint smears of 11 cases of MPE (mean age: 43.5 year‐old, 8 males and 3 females) were reviewed and graded semi‐quantitatively for 10 different cytomorphologic parameters. Cytologic examination revealed low to moderately cellular specimens composed of scattered isolated and loosely aggregated tumor cells with round to oval or occasionally spindle‐shaped nuclei, fine evenly distributed chromatin, and a meshwork of delicate processes. Predominantly observed cytologic features included a myxoid background, isolated single tumor cells, and long delicate cytoplasmic processes reflecting the glial nature of this tumor. Approximately half of cases contained “hyaline globules” (HGs) of varying sizes. Papillary structure, a histological hallmark of MPE, was rarely observed. No intracytoplasmic lumina, intranuclear inclusions or mitotic figures were seen. On cytomorphologic grounds, MPE may resemble EP and chordoma. Relatively lower cellularity of exfoliated tumor cells and a distinctive myxoid background with occasional HGs, and the presence of prominent delicate cytoplasmic processes supports a diagnosis of MPE rather than EP and chordoma, respectively.
Caveolin Inclusion Myopathy
Authors: Lach Boleslaw , Tarnopolsky Mark
Published: June 2021

We previously reported a new, autosomal dominant or sporadic, proximal myopathy characterized by mild myalgias and muscle weakness, and the presence of intracytoplasmic, hexagonally cross‐linked tubular arrays in type 2 muscle fibres (Ann Neurol 45:512, 1999). We report the ultrastructural and immunohistochemical characterization of identical inclusions in another patient with an apparent sporadic form of the disease.

This 45‐year old male underwent quadriceps femoris muscle biopsy for an upper extremity myalgias and CK activity found at 1,900 (N < 220 iU/L). He had no family history of muscle disease and parents were not consanguineous. Gomori stain revealed rod‐shaped intensely blue cytoplasmic inclusions in type 2 muscle fibres reminiscent of nemaline bodies. Inclusions were strongly positive for caveolin‐3 and negative for desmin, actin, myosin, vimentin, ubiquitin and components of sarcolemma. They displayed characteristic hexagonal crystalloid tubular arrays joined by double bridging arms and were not connected to cytoskeleton or any other organelles. The inclusions were very often surrounded by globular small structures measuring approximately 37 nm and reminiscent of caveolae. This is the first demonstration of chemical nature of the cytoplasmic inclusions in this condition. This disease is probably more common than reported, since the inclusions are very similar to nemaline bodies with light and electron microscopy, and misdiagnosed without grid tilting. At this point, it is not clear whether the underlying process is an excessive formation of caveolae or inability of cell to discharge or degrade caveolin, which conglomerate in the large intracytoplasmic masses.
RGS9‐2 differentially regulates adenylyl cyclase signaling by opioid and cannabinoid receptors in the mouse CNS
Authors: Selley Dana E. , Zachariou Venetia , Lichtman Aron H. , Nestler Eric J. , Sim‐Selley Laura J.
Published: June 2021

Regulators of G‐protein signaling (RGS) proteins accelerate G‐protein inactivation. RGS9‐2 is expressed in the CNS, and we have reported that this RGS regulates mu opioid receptor function in vivo. Here we report effects of RGS9 knockout on inhibition of adenylyl cyclase by opioid and cannabinoid receptors in the CNS. Results showed increased inhibition of adenylyl cyclase by the opioid partial agonist morphine, but not the full mu agonist DAMGO, in nucleus accumbens of RGS9 knockout compared to wild‐type mice. Similarly, inhibition by the cannabinoid agonist WIN55,212‐2 was increased in this region. Thus, RGS9 negatively regulates opioid and cannabinoid receptor signaling in a brain region important in drug reward. The periaqueductal gray (PAG) and spinal cord are two regions important in antinociception by these drugs. Mu opioid but not cannabinoid receptor‐mediated inhibition of adenylyl cyclase was significantly greater in PAG of RGS9 knockout mice, whereas inhibition by either receptor was not different in spinal cord. Antinociception testing showed greater sensitivity to morphine, but not the cannabinoid Δ9‐tetrahydrocannabinol, in RGS9 knockout mice. These results demonstrate differential effects of RGS9 knockout on opioid and cannabinoid receptor function in brain regions that mediate drug reward and antinociception. Supported by NID A.
Vascular inflammation in aging: role of increased mitochondrial H2O2 production in endothelial NF‐kB activation
Authors: Csiszar Anna , Labinskyy Nazar , Rivera Aracelie , Ungvari Zoltan
Published: June 2021

Aging elicits vascular oxidative stress and a pro‐inflammatory shift in endothelial gene expression profile. To elucidate the link between increased oxidative stress and vascular inflammation in aging the carotid arteries and aortas of young and aged (24 mo) F344 rats were compared. In aged vessels there was an increased NF‐kB activity (assessed by luciferase reporter gene assay and NF‐kB binding assay), which was attenuated by scavenging H2O2. Aging did not alter the vascular expression of p65 and p50 subunits of NF‐kB. In endothelial cells of aged vessels there was an increased production of H2O2, which was attenuated by the mitochondrial uncoupler FCCP. In young arteries and cultured endothelial cells antimycin A plus succinate significantly increased mitochondrial H2O2 generation, which was associated with activation of NF‐kB. In aged vessels inhibition of NF‐kB (by PDTC, resveratrol) significantly attenuated inflammatory gene expression and inhibited monocyte adhesiveness. Thus, increased mitochondrial oxidative stress contributes to endothelial NF‐kB activation, which contributes to the pro‐inflammatory phenotypic alterations in the aged vasculature.
Proliferative and apoptotic activities of homocysteine: new mechanisms of action in vascular smooth muscle cells
Authors: Rezk Bashir M , Reed Ryan E , Guidry Jesse J , Lucchesi Pamela A
Published: June 2021

Hyperhomocysteinemia is a risk factor for atherosclerosis. This study investigated molecular mechanisms by which homocysteine alters rat aortic vascular smooth muscle cell (VSMC) function. Low‐passage VSMC were treated for 24 hr with a pathophysiological level of homocysteine (150 μM). Paradoxically, homocysteine increased both VSMC proliferation and apoptosis. Proteomic analysis indicated that homocysteine induced a marked increased in cofilin and galectin‐1. Interestingly, homocysteine‐induced VSMC motility was associated with high expression of cofilin. On the other hand, immunocytochemistry results showed that the expression of galectin‐1 is co‐localized to apoptotic cells. The link between galectin‐1 and apoptosis was related to high expression of the proapoptotic Bax‐1. Conversely, the apoptotic effect of homocysteine involved activation of caspase‐3 and PARP cleavage. These results demonstrate for the first time that the proliferative and migratory responses to homocysteine were associated with high expression of cofilin, whereas the apoptotic pathway was associated with high levels of galectin‐1 and increased Bax‐1 expression, caspase‐3 activation and PARP cleavage to its active form. This novel finding may form a basis for new therapeutic strategies for treatment of atherosclerosis. Supported by HL056046 and P20RR18766 (PAL).
Adenosine anti‐adrenergic action in the rat heart is mediated in part through nitric oxide
Authors: Fenton Richard A. , Shea Lynne G. , Dobson James G.
Published: June 2021

Adenosine (ADO) is anti‐adrenergic in the heart (AJP 285:H1471, 2003). Nitric oxide (NO) has been reported to be anti‐adrenergic in rat ventricular myocytes. We studied the involvement of NO in the anti‐adrenergic action of ADO. 1) To determine if NO is released with ADO stimulation, an electrode measured NO in the culture medium of isolated rat cardiomyocytes. After 4 minutes of 100 μM ADO, the medium NO significantly increased by 12 nM. Additional experiments using the fluorescent NO indicator probe DAF‐FM loaded rat ventricular myocytes and a confocal system provided further evidence for ADO‐elicited NO release. 2) To determine if NO contributes to the anti‐adrenergic action of ADO, isolated rat hearts were perfused with 1 mM arginine. Contractility was enhanced with 25 nM isoproterenol (β‐adrenergic agonist). ADO at 10 μM reduced the contractile response by 25%, an effect that was significantly reduced by 1 μM L‐NMMA, a competitive inhibitor of nitric oxide synthase. 3) To determine if ADO alters NO synthase (NOS2) activity, ADO was infused at 10 μM into isolated rat hearts for 30 min, whereupon perfusion continued ADO‐free for 2 hours. Heart proteins were subsequently resolved, transferred and blotted with anti‐NOS2. An increase in the expression of NOS2 was observed. These data taken together suggest that ADO manifests an anti‐adrenergic action in the heart by a mechanism involving NO. (HL‐066045 & 084160)
The effects of non‐pressor and pressor doses of nitric oxide synthase inhibitor on renal excretion and regional blood flow in rat kidneys
Authors: Dobrowolski Leszek , Navar L Gabriel
Published: June 2021

While intrarenal administration of NO synthases inhibitors (NOSI) cause antinatriuresis, systemic infusion cause natriuresis (UNaV) which has been attributed to the associated increase in arterial pressure (AP). However, some studies show a time discrepancy between the rise in AP and UNaV. To evaluate the role of AP changes on renal function, the effects of L‐NAME (a nonselective NOSI), infused i.v., at non‐pressor (NPD, 5‐20 μg/kg/min) or pressor doses (PD, 50) were compared in anesthetized rats (n=12). UNaV was measured for 180 min together with inulin clearance (GFR) and renal cortical and medullary blood flows (CBF, MBF). The NPD increased UNaV after 150 min (0.7 ± 0.2 vs. 2.1 ± 0.4 μmol/min/g). The PD increased AP significantly after 30 min (115 ± 4 vs. 122 ± 4 mmHg), whereas UNaV increased after 60 min (1.1 ± 0.3 vs. 3.3 ± 0.7 μmol/min/g). GFR averaged 874 ± 44 μl/min/g and was not significantly altered by either NPD or PD. After 30 min of L‐NAME infusion, both NPD and PD reduced CBF and MBF. Moreover, the effects were dose related with CBF reduced by 17 ± 5% and 35 ± 6% and MBF by 27 ± 5% and 57 ± 7% for NPD and PD, respectively. Thus, NOSI effects on urinary sodium excretion effects are not directly related to changes in systemic arterial pressure and occur in presence of decreases in medullary and cortical blood flow.

Supported by NHLBI Grant R01HL‐18426
Manipulating the presence of hydrogen peroxide reveals redox modulation of rhythmogenesis originating from the in vitro pre‐Bötzinger complex.
Authors: Garcia Alfredo J. , Ramirez Jan‐Marino
Published: June 2021

The neuronal network within the pre‐Bötzinger complex (pre‐BötC) reconfigures itself to produce several patterns reflected in in vivo respiratory behaviors observed during changes in the oxygen environment. This study tests the hypothesis that changes in redox state surrounding the pre‐BötC has a role in network reconfiguration. Electrophysiological recordings were made in the pre‐BotC or hypoglossal nucleus of brainstem slices (~600um thick). Manipulating redox state with hydrogen peroxide (H2O2, 2.2mM) produced a depression followed augmentation in population burst frequency while extracellular catalase supplementation (20 Units/mL) produced augmentation followed by depression. Intracellular recordings from pharmacologically isolated respiratory neurons in the pre‐BötC indicate that modulating redox state, by altering the presence of H2O2, evokes changes in steady state membrane potential. Moreover, the effects on population burst frequency caused by H2O2 and catalase were also observed in the respiratory rhythm of the hypoglossal nucleus with the brainstem slice. These findings suggest that manipulating redox state of the pre‐BötC can influence inspiratory motor outflow. Our data indicate the redox state can act as a modulator of neuronal activity involved with respiratory drive. Supported Grant Number: T32HL007237 from the National Heart, Lung, and Blood Institute.
NADPH Oxidase Activation by Excess Uric Acid in Vitro
Authors: Yu Xiaohan , Zhang Suhua , Huang Harer , Hintze Thomas H.
Published: June 2021

Purpose: Uric acid is considered a major antioxidant thatmay protect against aging and oxidative stress. On the other hand, hyperuricemia induced alterations in oxidative homeostasis are commonly associated with increased risk for cardiovasculardisease and mortality. We hypothesized that acute elevated levels of uric acid induce O2− production by the activation of NADPH oxidase. Methods: SD male rats were divided into control(C) and Uric acid (UA) groups. The muscle tissues of left ventricle were incubated in Krebs solution without or with allopurinol, apocynin or tiron, 3mg/dL uric acid was placed in UA group, at 37°Cfor 1 h and bubbled continuously with 20% O2−5% CO2−75% N2. Results: NO‐dependent control of MVO2 stimulated by bradykinin or carbacholwas attenuated in UA (11.1% vs. 23.3% for BK; 13.2% vs. 20.9% for Cch). However, this was reversed by in the presence of apocynin (to 19.1% for BK; 19.6% for Cch) or tiron (to 20.6% for BK; 18.9% for Cch). It was not completely restored by allopurinol. Western blots of left ventricle showed that there was a significant difference between the two groups in the protein for p47 and nitrotyrosine (P < 0.01, P < 0.05 respectively), but there was no difference in eNOS, xanthine oxidase, p38 kinase, gp91, p22, SOD1 and SOD2. Conclusions: NADPH Oxidase was stimulated in acute hyperuricemia through the activation of p47. NO was inactivated by an increase in O2− production.

Supported by HL‐PO1‐43023 grant.
Effects of short and long‐term environmental air pollution exposure on antioxidant defense and sleep in adult mice
Authors: Abe Karina Camasmie , de Campos Brandão Letícia , Tufik Sérgio , Calegare Bruno Frederico Aguilar , Saldiva Paulo Hilário Nascimento , D′Almeida Vânia
Published: June 2021

It has largely been shown that air pollution can affect human health, but few studies have focused on effects on sleep parameters and antioxidant defense, aim of investigation of the present study. Male Swiss mice were placed in one of two chambers maintained inside them for 4 months until conception period. Mice were placed on the first chamber received air that had passed through an air filter (clean chamber) and the second received ambient air (polluted chamber), allowing us obtain groups with total exposure to air pollution, only gestational exposure and only gestational until weaning exposure. The experiments were carried out where the source of air pollution is predominantly automotive. Electrodes were implanted on animals permitting the assessment of the sleep–wake cycle. The activity of erythrocyte antioxidant enzymes ‐ catalase, glutathione peroxidase and superoxide dismutase ‐ were quantified. Differences on sleep parameters between groups were observed only in animals exposed all life on pollution and we observed a diminished in paradoxical sleep time when compared to control group (p<0.05). Moreover, the activity of all antioxidant enzymes, were decreased on mice exposed to air pollution during gestational period (p<0.05). The results suggest that air pollution could affect sleep parameters and was capable to decrease the erythrocyte antioxidant defense.

Finacial support: AFIP, FAPESP, CNPq.
Educating the Next Generation of Scientists: Lessons Learned from 17 Years of ASPET SURF Support
Authors: Bender Carol
Published: June 2021

The need for well‐trained scientists has never been greater. The biotechnology sector now accounts for more than a third of the world economy. What can be done to insure an adequate workforce?

The University of Arizona has received support from ASPET for Institutional Summer Undergraduate Research Fellowships (SURF) since 1991. This presentation will describe how our undergraduate research program has evolved over the past seventeen years and will provide information on outcomes. Suggestions will be given for those who wish to initiate similar activities with undergraduate students.

This work was funded by ASPET (no grant number).
Anatomical aspects with surgical value regarding the origin of the anterior tibial artery
Authors: MATUSZ Petru L.
Published: June 2021

The anterior tibial artery is a terminal branch of anterior bifurcation of the popliteal artery. The first part of its trajectory is deep into the popliteal fossa, in relation with the popliteus muscle. On a study material represented by 120 dissection specimens we analyzed the level and the modality of origin of the anterior tibial artery, as well as the particularities of its trajectory in the initial portion. Besides the modal type (present in 86.67% cases), where the origin of the anterior tibial artery is in relation with the inferior margin of the popliteus muscle and situated at 3–3.5 cm superior from the origin of the posterior tibial artery, in 13.33% cases we found major morphological variations of origin and branching of the anterior tibial artery trunk. In 9.17% cases we found a high origin of the anterior tibial artery (superior to the superior margin of the popliteus muscle). Within this group, in 0.83% cases the trajectory of the anterior tibial artery trunk is situated anterior to the body of the popliteus muscle, and in 2.5% cases the peroneal (fibular) artery originates from the trunk of the anterior tibial artery. In 4.17% cases, the anterior tibial artery has a common origin with the posterior tibial artery and the peroneal (fibular) artery (by trifurcation of the terminal portion of the popliteal artery), near the inferior margin of popliteus muscle. Knowing these morphological aspects of origin, trajectory and branching of the anterior tibial artery trunk represents the anatomical base for defining and delimiting calf and foot angiozomes.
Calpain involvement in monkey retinal cell damage due to hypoxia
Authors: Nakajima Emi , Shearer Thomas R. , Azuma Mitsuyoshi
Published: June 2021

One of the leading causes of blindness is retinal damage caused by the high intraocular pressure (IOP) in glaucoma. Previous studies in rats have suggested that the proteolytic enzyme calpain was involved in ganglion cells death during ischemia and acute high IOP. Glial cells are thought to protect neurons from various insults. For example, Müller cells, one type of glial cell in retina, are known to rescue ganglion cells from glutamate and NO neurotoxicities. The purpose of the present study using primate retina was to explain the role of calpains in hypoxic Müller cells. When dissociated cells from monkey retina were cultured for two weeks, Müller cells were observed with fewer photoreceptor cells, as identified by immunocytochemistry using vimentin and rhodopsin as markers. Under hypoxia, both types of cells were damaged, and a calpain‐specific α‐spectrin breakdown product (SBDP) was detected by immunocytochemistry. Activation of calpains 1 and 2, SBDP production and vimentin degradation were also observed by immunoblotting in hypoxic cells. When SNJ‐1945 calpain inhibitor was included during hypoxic culture, all the above changes were inhibited. The recovery of Müller cell damage by SNJ‐1945 might have important implications regarding amelioration of ganglion cell damage in glaucoma patients.

Dr. Shearer receives consulting fees from, and Dr. Azuma is an employee of, Senju Pharmaceutical Co. Ltd.
Kunitz Protease Inhibitor‐containing Amyloid Precursor Protein isoforms impaired mitochondrial function
Authors: Chua Li Min , Lee Chia Wen , Jacqueline  , Ho Zyet Ee , Wong Boon Seng
Published: June 2021

Reduced glucose metabolism is an early feature in Alzheimer's disease (AD), preceding the onset of the clinical symptoms of dementia and the appearance of the neuropathological amyloid‐beta (Aβ) deposits. Studies have shown that this metabolic deficit is associated with altered expression and activity of enzymes involved in mitochondrial energy production. However, the cause of this aberration is unclear. Aβ is proteolytically derived from amyloid precursor protein (APP), and there are three major splice forms of APP in the human brain; APP695, APP751,and APP770. In the brain, the predominant isoform transcript appears to be APP695, coupled with moderately high levels of APP751 and APP770. The APP751 and APP770 isoforms contain a Kunitz protease inhibitory (KPI) motif that APP695 lacks, and APP770 possesses an additional OX2 domain that is absent in both APP695 and APP751. In AD‐afflicted human brain tissues, APP751 and APP770 expression were elevated. In this study, we found that neuronal cell lines expressing KPI‐containing APP isoforms are associated with increased production of Aβ42 and impairment of mitochondrial function. In addition, the KPI‐APP isoforms have a greater propensity to co‐localize with mitochondrial‐specific protein markers. This subcellular localization may contribute to the observed aberrant mitochondrial function.
Phosphorylation and hetero‐oligomerization regulate the dopamine D3 receptor/adenylyl cyclase type 5 signaling complex
Authors: Yang Xiaodi , Sedaghat Keyvan , Plouffe Bianca , Gorji Hassan , Tiberi Mario
Published: June 2021

The dopamine D3 receptor (D3R) is a seven‐transmembrane‐spanning protein mediating its intracellular effects through the activation of heterotrimeric Gi/o proteins. The D3R‐mediated Gi/o protein activation culminates in an isoform‐specific inhibition of adenylyl cyclase type 5 (AC5) as previously demonstrated using a co‐transfection approach. In the present study, we explore the role of phosphorylation and hetero‐oligomerization in regulating the functional interplay between D3R and AC5 in co‐transfected human embryonic kidney 293 (HEK293) cells using immunoprecipitation, whole cell phosphorylation and cAMP assays. Whole cell phosphorylation experiments show that D3R exhibits basal and dopamine‐induced phosphorylation in cells not expressing AC5. Interestingly, our preliminary studies demonstrate that D3R is dephosphorylated in AC5‐expressing HEK293 cells following dopamine exposure. Moreover, our co‐immunoprecipitation studies imply that D3R and AC5 form hetero‐oligomers under basal conditions. However, dopamine stimulation curtails the D3R‐AC5 hetero‐oligomerization. Overall, our work suggests a novel paradigm in 7TMR‐mediated signaling whereby the phosphorylation of a 7TMR under steady‐state conditions and following agonist stimulation differentially regulate the hetero‐oligomerization process between the receptor and its effector. Supported by CIHR (MOP‐84442; MT).
Norovirus human infectivity, immunology, and persistence in groundwater
Authors: Leon Juan , McDaniels Marisa , Lyon G. Marshall , Abdulhafid Gwen , Dowd Melissa , Etienne Kizee , Liu Pengbo , Schwabb Kellogg , Moe Christine L
Published: June 2021

Noroviruses (NoV) have been responsible for outbreaks associated with contaminated ground, surface, and recreational water. To better understand the mechanism of waterborne NoV transmission, we examined how long NoV remains infectious in water. We seeded groundwater samples with high doses (1 × 107 genomic copies) of a safety‐tested Norwalk virus (NV) inoculum (8FIIb). We then challenged genetically susceptible volunteers (secretor positive) with this seeded water that was held at room temperature in the dark for various lengths of time. 13 volunteers received water with NV that had been stored for 0, 7, 14, 21, 27, and 61 days. Levels of NV RNA were stable in seeded water for over 61 days by quantitative real time RT‐PCR. All time‐points of seeded water induced infection (10/13) and illness in volunteers, including the water that was stored for 61 days. Duration of viral shedding varied by volunteer, and one volunteer asymptomatically shed NV for over 35 days. Only in infected volunteers did serum and salivary IgA and IgG levels increase after challenge. Inflammatory and anti‐inflammatory serum cytokines rose by day 4 post‐challenge in infected individuals. Our data suggest that NoV can remain viable in groundwater for over two months, can be shed asymptomatically by infected hosts for over 35 days post‐challenge, and pose a serious risk to untreated or contaminated groundwater. US‐EPA Grant 82911601‐0.
Pediatric residents‘ perceptions of barriers and needs for nutrition screening/counseling in well child visits
Authors: Lee Seung‐Yeon , Watkins Courtney , Gillengerten Rachele , Friedgen Shannon , McKinley Katie , Sigal Yakov , Hoerr Sharon
Published: June 2021

The purpose of this study was to elucidate residents‘ perceptions of barriers and needs for nutrition screening and counseling in well child visits. In‐depth interviews (n=12) were conducted with residents in a well child teaching clinic. Interview items probed for the well child visit routine, importance of nutrition/diet information, guidelines used, and barriers to nutrition screening/counseling. Content analysis was conducted on transcripts from audiotapes and field notes. Most residents believed that nutrition is an important part of the well child visit, in order to detect nutrition problems early enough to address health and developmental issues and to provide essential nutrition information to caregivers. Residents used direct questions and discussion with caregivers to obtain nutrition/diet information about their children. Major barriers included time constraints, language barriers, and lack of the caregivers‘ knowledge and receptiveness. Most residents wanted to have more dietitians at the clinic, to have a questionnaire for nutrition screening and age specific handouts/visual aids in the waiting/exam room. These findings will be used to develop nutrition screening/counseling protocols and handouts to improve the efficacy of nutrition care services in well child visits.

This study was supported by Families and Communities Together (FACT) Coalition at Michigan State University.
A free open source software for tracking tasks and the movement of pathology materials
Authors: Taieb Philippe , Deguzman Victoria C , Silver Steven S , Yong William H
Published: June 2021

An important function of a pathology laboratory is the tracking of blocks, slides, and tests to and from multiple locations. For research studies, a large group of cases may need to be tracked. Written logs are inefficient while bar‐coding and Radio Frequency Identification (RFID) technologies are costly and not widely available. Also, pathologists often track pending tests that might easily be lost to follow‐up. We have created a software program coupled to a Microsoft Access 2003 database and to an inexpensive digital camera that can track the ingress and egress of pathology materials as well as pathologist‐specific tasks. Destinations can be specified. Automated email reminders are sent when materials or pathologist‐specific tasks are due. When many cases are involved, the digital camera can image slides and blocks with sufficient resolution to capture the alphanumeric labels. One image can capture up to twenty slides in a tray or ninety blocks. Relevant documentation, such as shipping receipts, can be attached to the entry. A search function can find cases using specified parameters including name, destination, and date range. Hardcopy reports, containing all information regarding a case or group of cases, can also be created. This software can provide a relatively low cost, supplementary way to track processes and materials. It will be made available as free software under a GNU General Public License.
Genetic deletion of Regulators of G protein Signaling (RGS) protein activity enhances buprenorphine antinociception while limiting withdrawal behaviors associated with chronic administration
Authors: Talbot Jeffery N , Clemans Crystal F , Nicol Melanie R , Janjic Ana , Huang Xinyan , Neubig Richard R , Traynor John R
Published: June 2021

Understanding the mechanisms that govern opioid signaling is important to improve therapy for pain while limiting abuse potential. Opioid signaling is controlled by regulators of G protein signaling (RGS) proteins. In vitro evidence suggests that partial efficacy mu‐opioid agonists such as buprenorphine are more sensitive to RGS protein regulation than those that are fully efficacious. To determine the relevance of this in vivo, transgenic “knock‐in” mice were developed that express RGS‐insensitive Gαi2 (G184S; RGS‐i). Buprenorphine was twice as efficacious in RGS‐i animals compared to wild‐type littermates in antinociceptive tests. However, morphine activity was identical in animals of either genotype. In contrast, when administered chronically (5‐day treatment), both buprenorphine and morphine induced the same degree of tolerance in RGS‐i and wild‐type animals, indicating that mechanisms underlying opioid tolerance are insensitive to RGS regulation of Gαi2. Importantly, withdrawal behaviors, such as naltrexone‐precipitated jumping, were substantially reduced in chronically‐treated RGS‐i mice relative to wild‐type littermates. These data suggest that inhibition of RGS protein activity at Gαi2 leads to enhanced antinociception but reduced physical dependence to buprenorphine. Supported by DA04087, GM39561, T32 DA007268, and the Bower, Bennet, & Bennet Endowed Chair Research Award.
Direct inhibition of G protein signaling by cross‐conformational switches between α2A‐adrenergic and μ‐opioid receptors
Authors: Vilardaga Jean‐Pierre , Nikolaev Viacheslav , Lorenz Kristina , Ferrandon Sebastien , Lohse Martin J
Published: June 2021

Morphine, the powerful analgesic, and norepinephrine (NE), the principal neurotransmitter of sympathetic nerves, exert major inhibitory effects on peripheral as well as brain neurons by activating distinct cell surface G protein‐coupled receptors (GPCRs), the μ‐opioid (MOR) and α2A‐adrenergic receptors (α2AAR), respectively. Both receptors activate common signal transduction pathways mediated through the inhibitory G proteins (Gi and Go). Here we show that the MOR and α2AAR communicate with each other through a cross‐conformational switch that permits direct inhibition of one receptor by the other with sub‐second kinetics. We discovered that morphine binding to the MOR triggers a conformational change in the NE‐occupied α2AAR, which inhibits its signaling to Gi and downstream MAP kinase cascade. These data highlight a new mechanism in signal transduction at the cell surface whereby a heterodimer between different GPCRs mediated conformational changes that propagate from one receptor to the other and cause its inactivation.
The role of GPR30 in mediating the rapid vascular effects of aldosterone and estrogen
Authors: Gros Robert , Ding QingMing , Chorazcyzewski Jozef , Shaikh Rasha , Feldman Ross D
Published: June 2021

Aldosterone (ALDO) and estrogen (EST) elicit acute vascular effects through rapid signaling pathways. In vascular smooth muscle cells (VSMC), ALDO and EST mediate contraction. However, the mechanisms underlying this common effect remains unclear. Therefore, we examined the role of MR and GPR30 receptors in mediating ALDO and EST rapid effects in VSMC by assessing contractile response using the silicone wrinkling assay and ERK activation by western blotting. Short term (5min) incubations with ALDO or EST (1–1000 nM) mediated dose‐dependent VSMC contraction (ALDO:EC50=43±2 nM; EST:EC50=73±3 nM) and enhanced phenylephrine (PE)‐mediated contraction. The ALDO‐ and EST‐mediated increases in PE‐induced contraction were further enhanced with expression of either MR or GPR30. In contrast to their common effects mediating contraction, short term exposure to ALDO stimulated ERK activation (146 ±7 % of control), whereas EST inhibited ERK activation (67±3% of control), with further inhibition by expression of ERα. However, expression of GPR30 enhanced ALDO mediated‐ERK activation and reversed the effect of estrogen from ERK inhibition to ERK activation (149±11% of control). In summary, these data demonstrate that estrogen and aldosterone‐mediated vascular effects are mediated both by ER and MR, in part, but suggest an important role of GPR30 in mediating VSMC contraction as well as mediating ERK activation in VSMC.
Evidence that alpha1A‐adrenoceptors control blood pressure in isolated rat mesenteric bed and their relationship with mRNA.
Authors: Martínez‐Salas Sergio Gabriel , Sánchez‐Monroy Virginia , Campos‐Peralta José Manuel , Lozano‐Trenado Maritza Lucila , Villalobos‐Molina Rafael
Published: June 2021

Alpha1‐adrenoceptors participate in blood pressure control, where all three subtypes occur in blood vessels. However, it is known that one or two subtypes may be involved in arterial contraction. The aim of this work was to characterize the alpha1‐adrenoceptor subtype mediating muscular contraction in isolated mesenteric vascular bed and to determine the relative abundance of alpha1‐adrenoceptors mRNA.

Eight week‐old, male Wistar rats were used in the study, after anesthesia mesenteric vascular bed was isolated and connected to a blood pressure transducer (BIOPAC). The tissue was perfused with Krebs solution and either A61603 or phenylephrine as alpha1‐adrenoceptor agonists and in the presence of alpha1‐adrenoceptor antagonists (RS100329, chloroethylclonidine and BMY 7378). In other set of experiments RNA from different regions of the mesenteric be was obtained and real time PCR. Results shown that alpha1A‐adrenoceptor subtype predominates for blood pressure control while alpha1D‐adrenoceptor mRNA predominates in distal and proximal regions of the arterial tree, while alpha1B‐ subtype was not modified. Data suggest a negative relationship between functional and molecular expression of alpha1‐adrenoceptors in rat mesenteric vascular bed. It will be interesting to measure alpha1‐adrenoceptor proteins to explain this apparent discrepancy.
Pharmacokinetic studies with BLX‐1026, a novel Orexin‐2 receptor agonist with anti‐obesity property, in rats
Authors: Sen Ananda , Mukherjee Aparna , Neogi Partha , Dey Deben , Nag Abjijeet , Nag Biswajit
Published: June 2021

Orexin receptors have been implicated in non‐exercise activity thermogenesis. Increased levels of Orexin‐2 receptors have been observed in obesity‐resistant rats and decreased Orexin‐A levels observed in obese humans. BLX‐1026 is a water soluble, amino‐acid conjugated small molecule with in vitro Orexin‐2 receptor agonist activity. Phase I metabolism studies conducted with BLX‐1026 in rat, mice, dog, monkey and human liver microsomes and human liver S9 fractions show that it is stable in the liver microsomes but shows a 61% degradation in the human S9 fractions when incubated for 1 hour. In‐vivo pharmacokinetic study with BLX‐1026 was conducted in rats. It was administered at 50, 100 and 200 mg/kg, PO, and 10 mg/kg, IV, to male rats. Blood was collected for 24 hours and urine was collected for 72 hours following its administration. These were analyzed for the parent BLX‐1026 and any metabolites. The results will be presented and discussed.
Lung microvascular permeability determinations by two‐photon microscopy of single microvessels
Authors: Perlman Carrie E. , Lindert Jens , Bhattacharya Jahar
Published: June 2021

The lung microvascular barrier regulates transvascular macromolecular permeability (P) across the vascular surface area of filtration (S). The difficulty of P determinations lies in quantifying S. Hence, we addressed P in lung microvascular segments of fixed S. In the isolated, blood‐perfused rat lung held at constant left atrial pressure (Pla) of 3 cmH2O, we infused fluorescent dextran (20 kDa) in single venular capillaries. Since this Pla causes microvascular absorption of interstitial fluid (AJP 1993, 265:H198–204), we defined (1) diffusive capillary‐interstitial dextran flux, j, as j(t) = P[Cv(t)−Ci(t)] across constant S, where Cv and Ci are the luminal and interstitial dextran concentrations, respectively, and (2) Ci(t)=∫j(t)Sdt/V, where V is interstitial volume that receives transported dextran. To determine Cv and Ci, we quantified capillary and interstitial dextran fluorescence by two‐photon microscopy. Combining the above relations, we determined P for the capillary 2 min after the onset of dextran infusion. At baseline, P was 1.9±0.6 × 10−6 cm/sec (mean±SE, n=3). Following infusion of the permeability enhancer oleic acid (40 mg/ml, 10 min), P increased 121±26% (p<0.05). We conclude, our new method sensitively defines lung microvascular permeability at baseline and under barrier deteriorating conditions. (Support: HL80878, HL36024).
Nanomolar concentrations of ouabain stimulate Na‐K ATPase through sodium hydrogen exchanger‐1 (NHE1) dependent mechanism.
Authors: Holthouser Kristine , Chowhan Yaruk , Khundmiri Syed Jalal
Published: June 2021

Recent investigations have shown increased Na‐H exchanger activity and plasma levels of endogenous cardiotonic glycosides, such as ouabain in spontaneously hypertensive rats. At high concentrations, ouabain inhibits pump activity, and at lower concentrations, it increases pump activity and induces cell proliferation. Stimulation of NHE1 and Na‐K ATPase activity involve protein kinase B (AKT), suggesting that both NHE1 and Na‐K ATPase are mediated by the same proteins. We hypothesize that the activity of NHE1 and Na‐K ATPase are linked. In the present study we aim to 1) determine the role of activation of NHE1 in ouabain‐sensitive stimulation of Na‐K ATPase, 2) determine if NHE1 and Na‐K ATPase association increases when cells were treated with NH4Cl or ouabain in various kidney proximal tubule cell lines (OK, HK2, HK5, and HK11). Our data suggests that NH4Cl stimulation of NHE1 increases ouabain sensitive 86Rb uptake in all cell lines examined.Treatment with ethylisopropylamiloride (EIPA) a NHE1 specific inhibitor decreased Na‐K ATPase activity in HK5 and HK11 cells and prevented nanomolar ouabain‐stimulated Na‐K ATPase activity in OK cells. Immunoblot analysis indicated that NH4Cl increased membrane expression of NHE1 and Na‐K ATPase in OK and HK11 cells. Surprisingly, treatment with EIPA alone increased membrane expression of NHE1 and Na‐K ATPase. The reason for increased presence of these proteins in the membrane is unknown, however, tyrosine phosphorylation of the catalytic subunit of Na‐K ATPase, believed to be an indication of Na‐K ATPase activation, is decreased under these same conditions. These results suggest that activation of NHE1 is critical for ouabain stimulated increase in Na‐K ATPase activity, and that upon activation, NHE1 and Na‐K ATPase associate, perhaps to form a signalosome.