bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of a Supplementation with Ketogenic Amino Acids on Hepatic Steatosis Induced by Fructose in Healthy Humans
Authors: Theytaz Fanny Leila , Noguchi Yasushi , Egli Léonie , Campos Vanessa , Hodson Leanne , Patterson Bruce , Kreis Roland , Mittendorfer Bettina , Fielding Barbara , Boesch Chris , Tappy Luc
Published: June 2021

High protein intake blunts the deposition of intrahepatocellular lipids (IHCL) in fat‐overfed human volunteers. The aim of this study was to assess whether essential amino acids (EAA) supplementation also inhibits IHCL synthesis induced by a high fructose (HFr) diet.

9 healthy males were studied on 3 occasions, after 6 days of either: isocaloric balanced diet (C), the same diet supplemented with HFr or with HFr and EAA (HFrAA). IHCL content was measured by 1H nuclear magnetic spectroscopy and the effects of oral 13C fructose loads on VLDL‐13C palmitate production were monitored. HFr, compared to C, increased IHCL (2.7 ± 0.6 vs 1.3 ± 0.3 vol%) and plasma VLDL‐TG concentration (1.4 ± 0.2 vs 0.5 ± 0.1 mmol/L), both P < 0.05. It tended to increase VLDL production (16.7 ± 1.2 vs 12.0 ± 1.2 μmol/min), and enhanced VLDL‐13C palmitate production (195.2 ± 58.3 vs 4.0 ± 3.8 mmol/540 min, P < 0.05). Compared to HFr, HFrAA decreased IHCL (2.3 ± 0.5 vol%, P < 0.05), and further increased VLDL‐TG production to 21.2 ± 3.2 μmol/min (P < 0.05 vs C), but did not change VLDL‐13C palmitate production (326.6 ± 108.1 nmol/540 minutes).

We conclude that a supplementation with EAA significantly blunts the IHCL synthesis induced by fructose overfeeding, possibly by enhancing hepatic VLDL‐TG secretion.

Swiss National Science Foundation and Ajinomoto supported this work.
Oxalate contents in various types of tea drink
Authors: Pongthananikorn Suyanee , Mahanin Nunticha , Meksawan Kulwara , Tongyonk Linna , Panmuang Thitirat , Warisnoicharoen Warangkana
Published: June 2021

Oxalate is one of the major risk factors for kidney stones which become more common in industrialized countries. Tea is a significant source of oxalate. The purpose of the present study was to determine oxalate content in tea samples. One‐hundred and twenty samples of tea (white, green, oolong and black tea) were collected both in leaf and bag forms. The oxalate content was measured by the high performance liquid chromatography (HPLC) method with an anion exchange column using 0.85 mM sodium bicarbonate and 0.9 mM sodium carbonate as a mobile phase. The results showed that the highest of oxalate content was found in black tea (1.36–4.42 mg/g tea), followed by oolong tea (0.74–3.94 mg/g tea), green tea (0.44–2.18 mg/g tea) and the lowest was found in white tea (0.40–1.79 mg/g tea). The oxalate contents in leaf form and bag form were significantly different (p < 0.05). People in a high risk group should avoid drinking high amounts of black tea.
The effects of enriched oat milk supplementation on micronutrient status of Chinese schoolchildren
Authors: Zhang Huanmei , Zhang Qian , Hu Xiaoqee , Oste Rickard , Ma Guansheng
Published: June 2021

Oat milk has balanced macronutrient composition and free of lactose. In China, the lactose intolerance incidence is ca. 87.5% among young children and inadequacy of Ca, Fe, and vitamin D are common. To convey the benefits of oat milk to Chinese children, an oat milk formula was adopted in two 12‐mon randomly controlled intervention studies performed in rural Beijing. Its acceptance and effects of minerals and vitamins enriched oat milk on physical and mental development were evaluated. In one trial, total 409 children aged 8–10 were recruited from two schools and the study group was supplied with 200 ml Ca and Fe enriched product (A) twice/day/person. The other trial recruited 294 children aged 7–9 from a 3rd school. A carton of 250 ml Ca, Fe, vitamin A, and D enriched product (B) was served once/day. The subject compliance in both studies was ca. 91%. Product A was observed to promote bone mass accretion of children in the study group than those in the control, significantly higher e.g. at total body (162–180g vs. 147–167g). Children drinking product B gained significantly higher serum ferritin level. The studies showed that the enriched oat milk was accepted by most children. Oat milk enriched with suitable micronutrients may benefit mineral status.

The studies were financed by Oatly AB.
BMI, race, supplementation, season, and gestation affect vitamin D status in pregnancy in Kansas City (latitude 39° N)
Authors: Ozias Marlies K , Scholtz Susan A , Kerling Elizabeth H , Atwood Danielle N , Carlson Susan E
Published: June 2021

Numerous reports indicate a high prevalence of vitamin D deficiency in pregnancy. We asked whether vitamin D status changes during pregnancy and sought to identify predictors of status. Plasma 25‐hydroxycholecalciferol (25(OH)D) was measured before 20 wk gestation (n=299) and at delivery (n=257) in a mixed‐race population. Plasma was stored at −80° C until analysis. 25(OH)D was measured by EIA (Alpco Diagnostics, Salem, NH). Supplemental vitamin D was determined by capsule intake and brand. We used IOM guidelines for 25(OH)D: <12ng/ml (risk of deficiency), 12–19.9ng/ml (risk of insufficiency) and 20–50ng/ml (sufficiency). Status was evaluated for effects of race, pre‐pregnancy BMI, supplemental vitamin D and season of collection. At enrollment, 26, 30 and 41% were deficient, insufficient and sufficient. At delivery, 55, 30 and 15% fell into these respective categories. Plasma 25(OH)D decreased 40% from enrollment to delivery. Black women constituted 31% of the cohort but accounted for 69% of those deficient. Lower 25(OH)D was predicted by higher BMI at enrollment (p<0.001) and delivery (p<0.018). 25(OH)D at delivery was related to season of collection (p<0.001) and supplement intake (p<0.033). 25(OH)D declines in pregnancy and is sensitive to usual predictors in pregnant women residing in the Kansas City metropolitan area.
A prenylated flavan from Broussonetia kazinoki prevents cytokine‐induced ‐ [beta]cell death through suppression of NF‐[kappa]B activity
Authors: Lee Soyoon , Choi Younghwa , Lee Hwa Jin , Lee Da Yeon , Park Byung-Hyun , Ryu Jae-Ha
Published: June 2021

The generation of nitric oxide (NO) via inducible NO synthase (iNOS) and reactive oxygen species plays a key role in cytokine‐mediated pancreatic β‐cell damage. Oxidative stress due to reactive oxygen species activates the nuclear factor‐κB (NF‐κB) transcription factor, which regulates iNOS expression. In this regard, suppression of the NF‐κB pathway is a novel strategy for protecting β‐cells from damage. This study was performed to explore the effects of kazinol U, a prenylated flavan from Broussonetia kazinoki, on the NF‐κB activation pathway in IL‐1β‐ and IFN‐γ‐treated β‐cells. Kazinol U inhibited the nuclear translocation and DNA binding of NF‐κB subunits, which correlated with the inhibitory effects on IκB kinase (IKK) phosphorylation and IκBα degradation. In addition, kazinol U suppressed NO and hydrogen peroxide production and apoptotic cell death by cytokines in RINm5F cells. The protective effects of kazinol U were further demonstrated by normal insulin secretion of cytokine‐treated islets in response to glucose. Taken together, these results suggest that using kazinol U to block the NF‐κB pathway in pancreatic β‐cells reduces cell damage. Therefore, kazinol U may have therapeutic value in delaying pancreatic β‐cell destruction in type 1 diabetes.
Maturation specific sensitivity of osteocytes to calcium and phosphate ion pair‐induced apoptosis
Authors: Debo Matthew Paul , Saunders Ray , Genoese Angela , Adams Christopher Sean
Published: June 2021

Our previous work demonstrated the sensitivity of bone cells to microenvironmental apoptogens. As the lifespan of an osteocyte can extend into decades, osteocytic terminal differentiation must necessarily remove such sensitivity. Therefore, the goal of this study is to probe the relationship between osteocyte differentiation state and sensitivity to apoptosis. Murine Long bone‐derived osteocytes (MLO) A5 and Y4 cell lines, representative of early and late stage osteocytes, respectively, were seeded onto well plates, allowed to grow for 2, 7 or 14 days and subsequently exposed to increased levels of the calcium and phosphate ion pair for 24 h. Following exposure, MTT assays were performed to quantify cell death. Apoptosis was confirmed through caspase‐3 and TUNEL staining. Osteocytic phenotypes were confirmed through alkaline phosphatase staining. Results indicated that, when treated with increased levels of the calcium and phosphate ion pair, the MLO A5 cell line, representative of early osteocytes, exhibited greater sensitivity to apoptosis than the MLO Y4 cell line, representative of late osteocytes. These data suggest that sensitivity to calcium and phosphate induced apoptosis is a function of maturation level. This work was supported by a grant from the Center for Chronic Disorders of Aging at the Philadelphia College of Osteopathic Medicine.
Neutrophil recruitment in experimental sepsis: hypertonic solution action
Authors: Theobaldo Mariana , Llimona Flavia , Soriano Francisco Garcia
Published: June 2021

Sepsis is a systemic inflammatory response to infection and a major cause of mortality. Hypertonic saline solution (HSS) showed improvement on hemodynamic and inflammatory response. We studied the HSS effects in sepsis through cecal ligation and puncture (CLP) in mice Balb‐C, divided in 4 groups: CLP without treatment (CLP); CLP treated with saline solution 0,9% ‐ 34ml/Kg (SS); CLP treated with HSS 7,5% ‐ 4ml/Kg (SH); and a basal group without CLP and treatment; both treatments were applied 30 min after CLP. Lung and gut were collected after 6, 12 and 24h to analyze neutrophil recruitment using myeloperoxidase as index (MPO), ICAM, VCAM by ELISA and differential cell counter from peritoneal fluid. The SH group showed higher survival rate (60%) than SS (46,6%) and CLP (33,3%). CLP group (3 ± 0,4mU/mg; 29,4 ± 5,8 ng/mg) increased MPO and ICAM levels when compared to SH (1,5 ± 0,2 mU/mg; 15,9 ± 2,3 ng/mg) in lung at 24h. In gut MPO level increased in SH (0,9 ± 0,3mU/mg) when compared to CLP (0,1± 0,03mU/mg) at 6h; at 12h and 24h, SH decreased (0,2 ± 0,08; 0,2±0,05 mU/mg) in relation to CLP (3,5 ± 1,4; 0,6 ±0,05 mU/mg), ICAM levels in SS (1,7 ± 0,1 ng/mg) increased compared to SH (1,13±0,06ng/mg) at 6h, and SH (0,9±0,07ng/mg) decreased in relation to CLP (1,3 ± 0,07ng/mg) and SS (1,3 ± 0,1 ng/mg) at 12h. HSS protects the lung against tissue damage during sepsis, decreasing neutrophil recruitment into tissue through of a reduction in ICAM. CAPES.
Protease activated receptor‐1 (PAR1) control of glutamate time course in the synapse cleft
Authors: Traynelis Stephen F , Mannaioni Guido , Lee Jaekwong , Wahl Philip , Lee Changjoon Justin
Published: June 2021

The protease activated receptor‐1 (PAR1) is prominently expressed in astrocytes of the central nervous system. Activation of PAR1 increases the ability of synaptically‐released glutamate to surmount block by the rapidly unbinding competitive antagonist kynurenic acid. A variety of functional and molecular experiments suggest that this effect does not involve changes in presynaptic glutamate release or postsynaptic AMPA receptor function. Kinetic analysis of native AMPA receptors in outside‐out patches allowed the development of a simplified kinetic model of the actions of kynurenic acid. We subsequently used this model to fit the EPSC time course in the absence and presence of kynurenic acid with variable synaptic glutamate concentration profile. The result of this analysis showed that astrocytic G‐protein coupled receptor‐activation increased the peak synaptic glutamate concentration from 0.96 to 1.72 mM and prolonged the duration of glutamate in the synaptic cleft following PAR1 activation by approximately two‐fold. This effect was blocked by selective expression of a PAR1 shRNA in astrocytes. These data suggest that astrocytic G‐protein coupled receptors can influence the glutamate concentration time course in the synaptic cleft, which has important implications for excitatory synaptic transmission, as well as synaptic plasticity.
Melatonin inhibits SERT mediated 5‐HT uptake in human and mouse derived platelets and synaptosomes
Authors: Thompson Brent , Hill-Kapturczak Nathalie , Javors Martin
Published: June 2021

Melatonin is a hormone present in most organisms from bacteria to mammals. A circadian cycle of its release from the pineal gland into the blood and CSF is thought to be involved in a variety of normal and abnormal human behaviors such as sleep, epilepsy and autism. The serotonin transporter (SERT) can be found in multiple cell types including neurons, platelets, lymphocytes and intestinal epithelial cells where its principal function is to regulate the concentration of extracellular serotonin (5HT). Dysregulation of serotonergic signaling has been linked with numerous psychiatric and neurodevelopmental disorders. Melatonin has been shown to inhibit SERT function in a non‐competitive manner with a diurnal variation in responsiveness. Interestingly, a circadian rhythm of SERT function in human platelets has been shown to run counter to the concentration of melatonin in plasma leading to the hypothesis that melatonin may in part regulate SERT activity in human platelets and neurons. In these preliminary studies, we have characterized the effect of melatonin on 5HT uptake in platelets and synaptosomes derived from both human and mouse and have demonstrated that melatonin inhibits SERT mediated 5‐HT transport with inhibitory constants in the mM range.
Leptin anti‐apoptotic mechanisms in LPS‐induced PC12 cells
Authors: Masinde Sheila , Benzeroual Kenza E.
Published: June 2021

Evidence indicates that leptin has neuroprotective properties as it protects neurons from a variety of neurotoxic agents. However, leptin mechanisms have not been well studied. This study evaluated the anti‐apoptotic effects of leptin in LPS‐induced PC12 cells. Cells were pretreated with varying doses of leptin for 2 hours before stimulation with LPS for 48 hours. The results showed that LPS decreased cell viability, while pretreatment with leptin increased it in a dose‐dependent manner, as measured with MTT assay. Furthermore, western blot analysis showed a dose‐dependent reduction in the expression of cyclophilin D (CypD), an important component in the apoptosis pathway. The anti‐apoptotic effect of leptin was confirmed by the decrease in the expression of pro‐apoptotic protein Bad, and the increase in the expression of anti‐apoptotic protein BcL2. The present study indicates that leptin or its analogs, through inhibition of apoptosis may be promising novel therapeutic agents for the treatment of neurodegenerative diseases such as AD.
Translating heart failure biomarkers from animal models to humans
Authors: Altara Raffaele , Hessel Marleen M.H. , Manca Marco , Schmidt David , Hermans Rob J.J. , Janssen Ben J.A. , Blankesteijn Matthijs W.
Published: June 2021

The generation of a self‐evident paradigm where chemokines play a role in the remodeling of the heart is consistent with the idea that inflammatory molecules are responsible of the chemotactic recruitment and of a specific “remodeling oriented” inflammatory expression. In our in‐vivo experiments with animal models for heart failure (HF) such as transverse aortic constricted mice and myocardial infarcted rats we observed a common increased level of 4 novel (i.e. never investigated in this setting) circulating chemokines. The blood levels of >40 analytes were assessed semi‐quantitatively using a membrane‐bound antibody technique, called Proteome Profiler™(enhanced with fluorescence). The potential of the identified biomarkers in clinical studies, monitoring either new treatments or new strategies of clinical management, is capitalizing on recent advances in technology, bearing the promise of expediting the transition from “bench to bedside”, early diagnosis, individualized therapy and, as a consequence, better efficacy screening. In order to bridge the results obtained in the animal models with the clinic we proved the feasibility of the study using the blood of a small cohort of patients with heterogeneous HF. ELISA measurements plotted versus the membrane technique reached a linear correlation of R=0.93. These findings prompted us to begin a study in a larger cohort with established HF.
The effect of gender and age on hemodynamics, blood volume and cardiac size in healthy humans
Authors: Best Stuart , Okada Yoshiyuki , Galbreath M Melyn , Jarvis Sara S , Bivens Tiffany B , Levine Benjamin D , Fu Qi
Published: June 2021

We compared age and gender associated differences in hemodynamics, blood volume, and left ventricle size and function at rest in healthy sedentary adults. Ten young men [32 ±10 (SD) yrs] and 10 young women (30 ±8 yrs) as well as 12 older men (69 ±4 yrs) and 12 older women (70 ±5 yrs) volunteered for the study. Supine resting blood pressure (BP, Suntech), heart rate (HR, ECG) and cardiac output (Qc, C2H2) were measured. Stroke volume (SV=Qc/HR) and total peripheral resistance (TPR = mean BP/Qc) were calculated. Blood/plasma volume (BV/PV) was measured using a carbon monoxide rebreathing method. Cardiac size and function was assessed by MRI and later analyzed for left ventricular mass (LVM), left ventricle end diastolic volume (LVEDV) and ejection fraction (EF). BP and TPR increased, and normalized Qc, SV, BV, PV, LVM and LVEDV were significantly decreased in the older groups (all P<0.05). Females for both age groups had higher HR and TPR, lower BP, and lower normalized LVM and LVDEV than their respective male counterparts (all P<0.05). Normalized BV and PV were also lower in the female groups (both P<0.05). With age EF increased (P<0.05) but there was no gender effect on EF. This study shows both age and gender affect resting hemodynamics, blood volume, and cardiac size/function in humans, but the effect of gender is consistent with increasing chronological age.
Cardiovascular and respiratory responses to selected cranial osteopathic manipulations in healthy young adults
Authors: Ioudina Marina , Toder Eric , Zhang Frank , Syphus Dallin
Published: June 2021

Background and Purpose

Osteopathic manipulative treatment (OMT) for patient care may include cranial manipulations (CM). However, physiological responses to CM are not well defined. To study the effects of CM on the cardiovascular and respiratory systems the following measurements were used: heart rate (HR), HR‐variability, stroke volume index (SI), cardiac index (CI), and peak expiratory flow rate (PEFR).

Method

Subjects were healthy 25–35‐yr old men and women. PEFR was measured twice, at the beginning and at the end of a 30‐min protocol. Baseline cardiac parameters were obtained after a 10‐min rest in a supine position using impedance cardiography. Following CM that was performed by an osteopathic physician, cardiac responses were measured immediately and at the end of a 10‐min rest in supine position. Data were recorded and analyzed using BIOPACK Systems, Inc. equipment and software.

Results and Conclusion

Following CM, decreases were seen in HR (4%), CI (5%), and PEFR (10%) without significant changes in SI. The HR‐variability test revealed a decrease in sympathetic and increase in parasympathetic activity. These data demonstrate that the selected cranial osteopathic techniques decreased PEFR and decreased cardiac index, primarily by decreasing heart rate without a significant effect on cardiac contractility.
Exercise training reverses age‐related remodeling of coronary resistance arteries
Authors: Haynes Anthony Daniel , Hanna Mina , Chen Bei , Ghosh Payal , Delp Michael David , Delp Judy Muller
Published: June 2021

Age is known to induce remodeling and stiffening of large conduit arteries; however, little is known of the effects of age on remodeling and mechanical properties of resistance arterioles. We employed a rat model of aging to determine 1) whether age increases wall thickness and stiffness of coronary resistance arteries, and 2) whether exercise training reverses age‐induced increases in wall thickness and stiffness of coronary resistance arteries. Young (3 months) and old (21 months) Fischer 344 rats either remained sedentary or underwent 10 weeks of treadmill exercise training. Coronary resistance arteries were isolated for determination of wall:lumen ratio, elastic modulus, and active and passive responses to changes in intraluminal pressure. Wall:lumen ratio increased with age but this increase was reversed by exercise training. Similarly, age increased stiffness, whereas exercise training reduced it. Active responses to increasing intraluminal pressure (the myogenic response) were reduced with age and restored by exercise training. In contrast, passive responses to increasing pressure were not altered by age. These data suggest that age induces remodeling of the arteriolar wall, increasing the stiffness of coronary resistance arteries, and reducing the ability of the vascular smooth muscle to develop myogenic tone. Supported by NIH HL090937
ACE2 Deficiency Augments Cerebrovascular Dysfunction during Aging
Authors: Peña-Silva Ricardo Alfonso , Chu Yi , Mitchell Ian J , Miller Jordan D , Penninger Josef M , Faraci Frank M , Heistad Donald D
Published: June 2021

Angiotensin II produces oxidative stress and endothelial dysfunction. ACE2 converts angiotensin II to angiotensin 1–7 and may protect against effects of angiotensin II. We tested the hypothesis that ACE2 deficiency increases oxidative stress and vasomotor dysfunction in cerebral arteries, and examined the role of ACE2 in vascular aging. ACE2 was present at relatively low levels in cerebral arteries. Systolic arterial pressure was similar in adult and old ACE2 knockout (KO) and wild‐type (WT) mice. Maximal dilatation to acetylcholine (ACh) was impaired in the basilar artery from adult ACE2 KO mice compared to adult WT. In old mice, maximal dilatation to ACh was impaired in WT mice but severely impaired in ACE2 KO mice (P<0.05). The antioxidant tempol (1 mM) improved responses to ACh in adult and old ACE2 KO and WT mice. Nitrotyrosine staining in the basilar artery was increased in adult ACE2 KO mice and in old ACE2 KO and WT mice. Expression of NAD(P)H oxidase subunits Nox2 and p47phox, and of pro‐inflammatory molecules Rcan1 and TNFα, was increased in cerebral arteries from old ACE2 KO and WT animals. In summary, ACE2 deficiency impaired vasomotor function in cerebral arteries from adult mice and augments endothelial dysfunction during aging. Oxidative stress plays a critical role in cerebrovascular dysfunction induced by ACE2 deficiency and aging.

NIH grants NS‐24621, HL‐38091, HL‐62984; AHA 0815525G, 10PRE3780044.
Mitochondrial Uncoupling protein‐2 regulates NLRP3 inflammasome
Authors: Venugopal Rajan babu , Cox Ruan Rollin , Parthasarathy prasanna Tamarappu , Lockey Richard F , Kolliputi Narasaiah
Published: June 2021

Among the various types of inflammasomes, NLR family, pyrin domain containing 3 (NLRP3) senses a wide variety of stimuli. Inappropriate NLRP3 inflammasome activities have been reported in acute lung injury (ALI) and autoimmune diseases. Therefore, regulators of the NLRP3 inflammasome offer a considerable therapeutic promise. We investigate whether mitochondrial uncoupling protein (UCP)‐2 regulates inflammasome activation and modulates the functional effects of inflammasome mediated IL‐1β secretion. Differentiated human THP1 cells were pre‐treated with UCP‐2 inhibitor genipin followed by inflammasome activation with ATP. To mimic the infection model, THP1 cells were primed with LPS prior to genipin treatment. The expression of UCP‐2, NLRP3, ASC, pro‐caspase‐1 and pro‐IL‐1β in total cell lysates was assayed by western blot. Secretion of TNF‐α, IL‐6 and IL‐1β were detected by ELISA. We show that when activated THP1 cells are treated with genipin, the NLRP3 expression is suppressed. THP1 cells over expressed with UCP2 induced NLRP3 expression. Treatment with inflammasome stimulator ATP inhibits UCP2, but did not suppress NLRP3 expression. In addition, the presence of ATP restores genipin mediated suppression of NLRP3. In conclusion genipin inhibits NLRP3 through UCP2, but ATP mediated NLRP3 activation is independent of UCP‐2.

Research Supported by: R01 HL105932 (NIH) and 09SDG2260957 (AHA)
Granulocyte‐Colony Stimulating Factor Exerts Neuroprotection in the Neonatal Hypoxia‐Ischemia Rat Model via the Hypothalamic‐Pituitary‐Adrenal Axis
Authors: Charles Melissa , Ostrowski Robert , Manaenko Anatol , Duris Kamil , Zhang John , Tang Jiping
Published: June 2021

Neonatal hypoxia ischemia (HI) remains the leading cause of perinatal brain injury affecting 60% of preterm infants. Several reports suggest that the activity of the hypothalamic‐pituitary‐adrenal (HPA) axis is increased after HI. Granulocyte‐colony‐stimulating factor (G‐CSF) is a neuroprotective drug shown to modulate the HPA axis in the adult rat by regulating the adrenocorticotropic hormone (ACTH) and corticosterone (CORT). We hypothesize that G‐CSF confers neuroprotection in neonatal HI by suppressing the activity of the HPA axis. To test the hypothesis Metyrapone (MET), a CORT synthesis inhibitor, and dexamethasone (DEX,) a synthetic glucocorticoid, was administered separately or in combination with G‐CSF. HI was induced in P7 rats following a modified Rice‐Vanucci model, which were randomly assigned to groups and treated one hour after HI. We report that G‐CSF is neuroprotective in neonatal HI by reducing infarct volume, and regulating CORT levels. The combination of G‐CSF and MET increased weight gain, and significantly decreased pro‐apoptotic markers in the brain; while DEX reversed the effects of G‐CSF.
Whole body surface cooling before aerobic exercise decreases lactate threshold in cool environment
Authors: Imai Daiki , Okazaki Kazunobu , Suzuki Akina , Takeda Ryosuke , Naghavi Nooshin , Matsumura Shinya , Miyagawa Toshiaki
Published: June 2021

Whole body surface cooling prior to exercise improves aerobic performance in the heat, while possible effects of the procedure on aerobic performance in cool environment are remained unknown. Sympathetic nerve activity and glycolysis are enhanced by body surface cooling, therefore we hypothesized that whole body surface cooling prior to exercise increased plasma lactate concentration [Lac]p before and during aerobic exercise and declined lactate threshold (LT) in cool environment.

Methods

Six healthy young women and men performed a graded maximal exercise on a cycling ergometer after pre‐conditioning (60 min) in 3 conditions. The ambient temperature (Ta) and a water perfusion suit temperature (Tw) were controlled at Ta10°C‐Tw10°C in Cool‐Cool (CC), Ta25°C‐Tw10°C in Mild‐Cool (MC), Ta25°C‐Tw34°C in Mild‐Neutral (MN), respectively. [Lac]p, heart rate, blood pressure, expired gas, esophageal (Tes) and skin temperature (Tsk) were measured and LT was determined.

Results

Tes was higher and Tsk was lower in CC and MC than MN after pre‐conditioning, respectively. [Lac]p after pre‐conditioning and during each workload of exercise were higher in the order of CC, MC, MN, while LT showed lower values in the same order. [Lac]p at exhaustion was the lowest in CC.

Conclusions

Whole Body surface cooling before aerobic exercise increased [Lac]p before and during exercise and decreased LT in cool environment.
Effect of AMPK activation on acute exercise‐induced monocarboxylate transporter (MCT) 1 and MCT4 mRNA expression in skeletal muscle
Authors: Takimoto Masaki , Takeyama Mirei , Enoki Taisuke , Hamada Taku
Published: June 2021

We examined the mechanism on acute exercise‐induced MCT1 and MCT4 mRNA expression in skeletal muscle. The rats were subjected to a bout of high‐intensity intermittent swimming (HIS) or low‐intensity prolonged swimming exercise (LIS). The HIS rats swam for fifteen 20‐second with a weight equal to 18% of their body weight. The LIS rats swam without load in two 3h sessions separated by 45min of rest. HIS elevated the phosphorylation of 5 fAMP‐activated prtein kinase (pAMPK), calcium/calmodulin‐dependent kinase II (pCaMKII), and p38 mitogen‐activated protein kinase (pMAPK), which are characterized upstream modulators of transcriptional regulation. LIS elevated pAMPK, whereas pCaMKII and pMAPK were not altered. MCT1 and MCT4 mRNA were transiently upregulated by HIS and LIS. Direct exposure of epitrochlearis to 0.5mM 5‐aminoimidazole‐4‐carboxamide‐1‐ƒÀ‐d‐ribonucleoside (AICAR) or 1mM caffeine increased both MCT1 and MCT4 mRNA. When 1mM caffeine‐induced pAMPK was inhibited by dantrolene, neither MCT1 nor MCT4 mRNA was increased. The present findings suggest that, irrespective of exercise intensity, acute exercise may upregulate MCT1 and MCT4 mRNA expression, through AMPK‐mediated pathway in skeletal muscle.
Imaging of muscle blood volume changes during exercise with Vascular Space Occupancy (VASO) MRI
Authors: Godar Jared A , Rane Swati , Towse Theodore F. , Damon Bruce M.
Published: June 2021

Large, rapid changes in perfusion accompany exercise, mediated in part by a local vasodilatory response. Impaired vasodilation occurs in diabetes, obesity, and other conditions. VASO MRI was developed to measure blood volume changes in the brain by nulling the signal of blood; vasodilation thus causes a loss of MRI signal intensity. The purpose of this work was to implement VASO for skeletal muscle. Eighteen subjects participated in the optimization of this method. Subjects were placed supine in a Philips 3.0T scanner with their foot on a force transducer, a knee coil around the calf, and an occlusion cuff around the thigh. Anatomical and VASO images were acquired (Fig 1).

Maximal plantarflexion contractions were performed for 1–20 seconds, six minutes apart. The cuff was inflated to 240 mm Hg for five minutes, occluding arterial flow; VASO response was monitored for up to fifteen minutes after cuff release. NIRS spectroscopy was also performed to relate observed VASO signal changes to blood oxygenation and volume changes. VASO signal follows a similar time course to NIRS post occlusion.

The decrease in VASO signal following a series of three ten second maximal plantarflexion contractions, as shown in Fig 2, should be proportional to increased blood volume demonstrating the promise of this new imaging paradigm to study blood volume changes in exercising muscle. This work was supported by NIH/NIAMS R01 AR050101.
Endogenous hydrogen sulfide is involved in the brain's control of arterial blood pressure
Authors: Sikora Mariusz , Ufnal Marcin
Published: June 2021

Hydrogen sulfide (H2S) has recently been found to play an important role in the regulation of the circulatory system, however, its effect on the brain's control of arterial blood pressure has not been established yet. In the study we investigated the hemodynamic effects of intracerebroventricular (icv) infusions of S‐adenosyl‐L‐metionine (SAM), an activator of cystathionine betasynthase which produces H2S in the brain. We preformed experiments on freely moving, male, Wistar Kyoto rats. Mean arterial blood pressure (MAP) and heart rate (HR) were recorded at baseline and during icv infusions of SAM. The infusions produced a dose dependent, biphasic response, i.e. moderate hypotension and bradycardia lasting 10–15 min, which were followed by long‐lasting significant increases in MAP and HR. These findings suggest that endogenous H2S in the brain is involved in the neurogenic control of blood pressure, and that the effects of H2S may depend on its concentration and/or the site of its action in the brain. This work was supported by the National Science Center, grant 3G29 and Medical University of Warsaw, grant 1MA/PM11/11.
Early‐stage changes to neurovascular morphology and telemetric measurement of sympathetic nerve activity in streptozotocin‐induced diabetic rats
Authors: Grise Kenneth Neil , Hall Katharine E , McDonald Matthew W , Noble Earl G , Melling C.W. James
Published: June 2021

Chronic sympathetic nerve overactivity is an established consequent of Type 1 Diabetes Mellitus (T1DM) and is known to induce both micro and macroangiopathy. Telemetry is used to monitor real‐time sympathetic nerve activity (SNA) to measure neuropathic progression in T1DM. This study aimed to examine changes in the neurovascular morphology of early‐stage streptozotocin (STZ)‐induced T1DM rats and determine if telemetry is sensitive to neuropathic changes in SNA. Animals were separated into control (C, n=9) and T1DM (D, n=9) groups. Diabetes was induced using multiple doses of STZ consisting of 20mg/Kg per day for 5 days. In three animals from each group, electrodes were placed around isolated renal nerves and telemeters were implanted abdominally. Measurements of SNA were recorded weekly over 12 hour periods. All animals were euthanized 6 weeks post STZ treatment. Compared to C, D showed a decrease in blood vessel number (t (10) = 4.44, p < 0.001) and size (t (10) = 3.60, p < 0.05), an increase in sympathetic fiber size (t (9) = –2.34, p < 0.05) but no difference in fiber number in sciatic nerves. Despite these morphological changes, SNA in C and D was not statistically different in frequency or amplitude at any time‐point during the 6 week period. Though morphological changes are observed early in STZ diabetes, neurophysiological changes recorded by telemetry may be too slight or variable to be clearly distinguished.
Renovascular remodeling in Angiotensin‐II induced hypertension is strain–dependent
Authors: Pushpakumar Sathnur , Kundu Sourav , Givvimani Srikanth , Tyagi Suresh , Sen Utpal
Published: June 2021

Hypertension is a recognized cause of chronic kidney disease and end‐stage renal dysfunction. Sustained hypertension induces renovascular remodeling in both intra and extra‐renal vasculature and alters extracellular matrix (ECM) components. Our objective was to define renovascular phenotypes in agonist induced hypertension in different strains of mice. Osmotic pumps loaded with Angiotensin‐II (Ang‐II) were inserted subcutaneously into C57BL/6J (WT), TIMP2−/−, and C3H/HeJ mice for delivery at 250 ng. kg‐1. min‐1.

Results

Ang‐II induced higher blood pressures in WT and TIMP2−/− mice compared to C3H mice. Blood flow across renal artery and aorta was decreased by 33% in TIMP2−/− mice; whereas, in WT and C3H mice, flow decreased by 23 and 20% (aorta) and 19 and 10% (renal artery) compared to untreated controls. Barium microangiography showed decreased renal vascular density in TIMP2−/− and WT groups than in C3H. Peri‐glomerular and vascular collagen deposition was increased in TIMP2−/− and WT compared to C3H. Elastin deposition, however, was predominant in WT than in the other groups. Intracellular reactive oxygen species production was low in C3H mice, increased moderately in WT and was twice as high in TIMP2−/− than in C3H.

Conclusions

Our results suggest that in Ang‐II induced hypertension, renovascular response is strain dependent indicating a genetic basis for disease susceptibility.
Rat Kidney biomarkers: 613‐APS (renal)
Authors: Schmidt Jane
Published: June 2021

Multiplex measurement of rat kidney cytokines in serum and urine using the Fluorokine® MAP assay

Jane Schmidt Ph.D, Michael Rynning, Richard Fuerstenberg R&D Systems, Inc., Minneapolis, MN

Drug toxicity can result in major organ damage to the kidneys. The traditional methods for monitoring drug‐induced nephrotoxicity in toxicology studies include measurement of serum creatinine and blood urea nitrogen levels, but in recent years more sensitive kidney biomarkers have emerged. The PCK rat is a recently identified rodent model of polycystic kidney disease that can be used as a model for general kidney disease or damage. We measured TIM‐1/KIM‐1, FABP‐1, Lipocalin‐2, Cystatin C, and Osteopontin in serum and urine from normal (Sprague Dawley) and PCK rats using the Fluorokine MAP Rat Kidney Toxicity Panel and observed a significant increase of TIM‐1/KIM‐1 and Lipocalin‐2 in the PCK rat serum compared to normal rat serum. We also observed statistically significant differences for Lipocalin‐2 and Cystatin C in PCK urine when compared to normal urine. Fluorokine Multianalyte Profiling (MAP) kits are bead‐based multiplex immunoassays designed for use on Luminex® 100/200™, Bio‐Rad® Bio‐Plex® analyzers and offer the ability to quantify multiple analytes simultaneously with less user time, less sample volume, and lower cost per result compared to conventional ELISAs. Five biomarkers can be measured in a user‐defined multiplex using the Rat Kidney Toxicity Panel. The assay is fully validated for accurate, precise, and reproducible results in rat serum, plasma, and urine.
Intermittent hyperoxia attenuates the hypoxic ventilatory response in neonatal rats
Authors: Tobin Kristina E , Bavis Ryan W
Published: June 2021

Sustained exposure to 30–60% O2 from birth reduces the hypoxic ventilatory response (HVR) in rats, primarily through abnormal development of the carotid body. We hypothesized that a more clinically relevant intermittent hyperoxia exposure would produce similar changes to the HVR. Rats were continuously exposed to intermittent 30% O2 or intermittent 60% O2 from birth through 14–15 days of age (i.e., 4.5‐min bouts of hyperoxia separated by 7.5 min of normoxia, 24 h day−1); Control rats were reared in room air. Ventilation was measured at 14–15 days of age by head‐body plethysmography (n=15–18 per group). No differences were detected among treatment groups while breathing 21% O2. However, ventilation was significantly lower in both groups of intermittent hyperoxia rats during acute exposure to 12% O2 (143±8 and 154±4 ml min−1 100g−1, respectively) than in Control rats (172±6 ml min−1 100g−1; both P<0.05). Although attenuation of the HVR was modest relative to that observed after similar durations of sustained hyperoxia, these data indicate that intermittent exposure to hyperoxia can alter the normal development of ventilatory chemoreflexes. Supported in part by NIH grant P20 RR‐016463 (Maine INBRE).
Identification of the DNA Binding Surface on the SIRV Capsid Protein
Authors: Allgaier Hannah E , Taurog Rebecca E. , Johnson John E. , Rohlman Christopher E , Szymczyna Blair R.
Published: June 2021

Viruses that infect archaea have a unique system of resilient capsid proteins that must assemble and protect the virus genome from the harsh conditions in which some of these organisms survive. The Sulfolobus islandicus rod shaped virus (SIRV), for instance, is a non‐enveloped virus of the Rudiviridae family that infects the thermophilic and acidophilic Sulfolobus archaea. Members of this virus family have been isolated from solfataric fields in Iceland, Italy and the United States (Yellowstone National Park). Since the capsid of the Rudiviridae viruses is resilient to high temperatures and low pH, the capsid structure has potential uses as a nanobuilding block in biotechnological applications. An understanding of the requirements for SIRV capsid assembly will aid in the design of such nanomaterials. The solution structure of the C‐terminal domain of the SIRV capsid protein reveals the domain to be a four‐helix bundle. We are currently employing solution state NMR techniques to investigate how this region of the capsid protein associates with the double stranded DNA genome, and to ultimately get clues about the mechanism of capsid assembly.
The phage e34 repressor binds to a unique operator
Authors: Williams Jeremie Durrell , Jackson Doba , Villafane Robert
Published: June 2021

The goals of this study are to: 1) compare the operator‐promoter sites of the Salmonella newington phage e34 lysogenic repressor with those same elements of the Salmonella typhimurium phage P22; 2) compare the amino acid sequence and structure of this e34 phage protein with those same elements of the Salmonella typhimurium phage P22; 3) determine the operator sequences of the e34 operator site(s). Towards this end, the genome of the entire phage was sequenced. The DNA of the repressor gene was cloned into a pET11a vector. DNA from a small number of clear plaque variants was sequenced. Our results indicate that the operator‐promoter sequence of e34 operator site is unlike that of the corresponding regions of P22 and lambda phage. It does not contain the characteristic three seventeen base operator boxes. Furthermore, the N‐terminal domain (NTD) of the e34 repressor is not similar to that of either of these prototype phages. The expectation is that this phage will yield a new type of repressor binding site motif. The NTD domain of the e34 repressor contains a typical helix‐turn‐helix DNA binding motif and two mutations, that have been isolated, are located on a conserved phenylalanine residue just seven amino acids away from the putative recognition helix.
CYP2D6 is the Major Metabolizing Enzyme of Metoclopramide
Authors: Briggs Errran D. , Bolles Amanda K. , Livezey Mara R. , Furge Laura Lowe
Published: June 2021

CYP2D6 metabolizes approximately 20% of pharmaceutical drugs. The important role that CYP2D6 plays in drug metabolism makes inactivation events clinically relevant. Thus, understanding the inactivation of CYP2D6 may prevent adverse drug‐drug interactions. Metoclopramide, a drug commonly prescribed to counteract nausea in chemotherapy patients, has previously been reported in the literature to be a mechanism‐based inhibitor of CYP2D6. We sought to expand this initial finding by constructing a metabolite profile and employing molecular modeling programs in an attempt to better understand the interactions between metoclopramide and CYP2D6. While we did not observe mechanism‐based inhibition with metoclopramide, we did observe predicted and new metabolites of metoclopramide. Furthermore, we were able to establish the relative contribution of specific P450 enzymes to the metabolic profile of metoclopramide and the role of CYP2D6 as the major metabolizer of metoclopramide. (Support: NIH 1R15‐GM086767–01 & ‐01S1).
Synthesis of Novel Pb, Mn, Zn, Ag, Co and Cu Complexes With Coumarine Derivative Moiety and Their Antibacterial Activities
Authors: Govori Sevdije R , Haziri Arben , Palucaj Fabijan , Krasniqi Agron , Rifati Arleta , Ibrahimi Hamide
Published: June 2021

Coumarine derivatives often show significant antibacterial activities. A number of coumarine metal complexes have a wide range of biological activity. In order to find new biological active coumarine derivatives, we prepared new coumarin complexes of Pb, Mn, Zn, Ag, Co and Cu (2–7) with 3‐nitro‐4‐(heteroaryl‐2‐ ylamino)‐coumarine derivatives 1, in very good yields. Synthesized coumarin metal complexes (2–7) were investigated for antibacterial activities using agar diffusion method (in vitro) against Staphylococcus aureus and Escherichia coli. Applying the disc method, we measured diameters of the inhibition zone around disc which are previously wetted with N,N‐DMF solution of compounds (2 – 7) in concentration 1 mg/mL, 3 mg/mL and 5 mg/mL. These new synthesized compounds showed moderate activity in reducing the microbial growth (6.5–13 mm). Comparing the inhibitory activities of the tested complexes, it was found that these complexes were a bit more active than coumarine compound 1.
Postnatal injection of homocysteic acid in rats leads to the development of schizophrenia‐like behaviors
Authors: Flores Guillermo , Barney Christopher , Chase Leah
Published: June 2021

Hypofunction of N‐methyl‐D‐aspartate receptors (NMDA) in GABAergic interneurons is likely an important factor in the development of schizophrenia. The NMDA receptor agonist, homocysteic acid (HCA), is known to cause cytotoxicity in NMDA‐receptor containing neurons. In addition, HCA is a substrate for the cystine/glutamate exchanger, system xc−. While system xc− has broad distribution, it is known to be expressed at the blood brain barrier and GABAergic interneurons of the hippocampus. We hypothesized HCA may be selectively concentrated in GABAergic interneurons by System xc−, thus increasing the susceptibility of these interneurons to NMDA‐mediated excitotoxicity by HCA. To test this hypothesis, we performed daily intraperitoneal (IP) injections of either HCA or saline in postnatal rats for 14 days beginning at day P2. Six weeks later, we observed HCA‐treated rats displayed increased risk‐taking behavior, mild anhedonia, deficits in prepulse inhibition, and increased sensitivity to pain compared to control rats, but no difference in motor performance or weight gain over the course of the experiment. These data suggest that exposure to HCA during early development leads to behavioral changes that are consistent with a schizophrenia phenotype. This research was supported by the Hope College Neuroscience Program, Biology Department and Chemistry Department, and NSF‐REU # 0851194.
Expression, extraction and activity testing of the metacaspase Scp3
Authors: Gagliardi Connor , Fox Kristin M
Published: June 2021

Apoptosis, a fundamental characteristic of multicellular eukaryotic organisms, is a key component in maintaining organismal integrity. Proteins called metacaspases regulate apoptosis in plants, protozoans and fungi. These proteases are responsible for cleavage of other proteins during programmed cell death. Metacaspases preferentially cleave after basic residues in the P1 subsite. The substrate specificity at the P2 and P3 subsites varies for the metacaspases in plants and fungi. But, metacaspase specificity been studied in only a few plants and in two fungal organisms (S. cerevisiae and S. pombe). Our lab has been studying the five metacaspases from Schizophyllum commune, a split gill mushroom, to better understand the substrate specificity and function of these enzymes. The metacaspase Scp3 has been successfully cloned from the S. commune genome, and transformed into and expressed from Escherichia coli. Using two rounds of nickel affinity chromatography we have obtained protein that is >;95% pure. Current activity assays have shown significant enzymatic activity of Scp3 with the substrate FITC‐Casein compared to a crude cell extract without Scp3. Our lab is currently working to optimize the expression of Scp3 via autoinduction in various media and at different growth temperatures. Additionally, optimization of the extraction and assay conditions for the metacaspase are being completed in conjunction with one another. Optimization of extraction will provide us with more stable and active protein that can be tested for enzymatic activity. Once optimization of all processes is completed for Scp3 the knowledge can be transferred to the other four metacaspases of S. commune.
Two domains of the smoothelin‐like 1 protein bind apo‐and calcium‐calmodulin independently
Authors: Ulke‐Lemee Annegret , Ishida Hiroaki , Vogel Hans J , MacDonald Justin A
Published: June 2021

The smoothelin‐like 1 protein (SMTNL1; also known as calponin homology (CH) domain‐associated smooth muscle, CHASM) is a modulator of vascular smooth muscle contractility and can bind to calmodulin (CaM) and tropomyosin. The intracellular calcium receptor CaM switches between calcium‐free (apo‐CaM) and calcium‐bound (Ca‐CaM) forms. Bioinformatic analysis of the SMTNL1 sequence predicted two CaM‐binding regions: CBD1, N‐terminal to the CH domain and CBD2, a previously described IQ motif within the CH domain. We sought to describe the contribution of SMTNL1's two CBDs to CaM binding in the presence or absence of calcium. Using a variety of assays (biochemical pull‐downs, surface plasmon resonance, NMR and intracellular co‐localization by microscopy), we determined that CBD1 binds preferentially to Ca‐CaM while CBD2 binds preferentially to apo‐CaM. Mutation of key acidic residues abolished apo‐CaM binding to CBD2, while hydrophobic residues in CBD1 were required for Ca‐CaM binding. The binding affinities (~4 ×10−6 M) were similar for both. The association of SMTNL1 with apo‐CaM and Ca‐CaM suggest that endogenous SMTNL1 is always associated with CaM. This lead us to the conclusion that changes in the CaM binding to CBD1 or CBD2 may allow for downsteam effects, such as disruption of SMTNL1 binding to tropomyosin.