bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of temperature, anesthesia, and infection on urinary bladder pain‐like responses in mice
Authors: Sadler Katelyn Elizabeth , Stratton Jarred Michael , Kolber Benedict J
Published: June 2021

Painful bladder syndrome/interstitial cystitis (PBS/IC) is a condition characterized by chronic bladder pain. Patients suffering from PBS/IC are commonly unresponsive to regular pain treatment and current models of bladder pain in animals do not recapitulate many of the important features. We sought to improve this situation through the use of urinary tract infections (UTI) in mice to model the bladder sensitization seen in human patients. In our new system, following UTI, urinary bladder distension (UBD) was combined with visceromotor response (VMR) measurements to evaluate the pain‐like response to bladder distension. In designing this model we discovered several potentially confounding variables such as body temperature and length of isoflurane anesthesia. We found that UBD‐induced VMRs decrease as body temperature decreases. These data suggest that body temperature must be closely controlled during UBD testing. In addition, we found a significant effect related to the length of time under anesthesia. When mice were tested with anesthesia times under two hours, VMR was reduced with increasing length of anesthesia time. Interestingly, after two hours no significant decrease in VMR was seen with increasing length of anesthesia. After controlling for these variables, preliminary data was gathered on the effect of UTI on VMR. The data suggests a modulation of VMR following UTI. Support provided by APS UGSRF.
3‐[2‐(3,5‐dimethoxyphenyl)vinyl]furan protects hippocampal neurons against ischemic damage
Authors: Choi Sang Yoon , Hur Jinyoung , Cho Chang‐Won , Kim Sanghee
Published: June 2021

Resveratrol, an ingredient in grapes, has been reported to exhibit anti‐cancer activity, anti‐inflammatory activity, and cardiovascular protection property. Interestingly, resveratrol has been recently reported to have neuroprotective effect. This study reports the neuroprotective effect of a resveratrol derivative, 3‐[2‐(3,5‐ Dimethoxy‐phenyl)‐vinyl]‐furan (DPVF). This synthetic DPVF conferred more protection than resveratrol against neuronal cell damage induced by oxygen and glucose deprivation in a rat hippocampal slice culture. In addition, DPVF inhibited ATP depletion following oxygen and glucose deprivation in the adult hippocampal slice. Moreover, we found that DPVF is neuroprotective against ischemic damage in rats. DPVF showed potent neuroprotection on a 4‐velssel‐occusion model and inhibited iron‐induced malondialdehyde (MDA) formation in the rat brain tissue. These results demonstrate that DPVF might be a useful agent in reducing ischemic neuronal damage.
Renin‐angiotensin system potentiates inflammatory responses of microglia
Authors: Shen Xiao , Bernstein Kenneth B , Lyden Patrick , Shi Peng
Published: June 2021

Renin‐angiotensin system (RAS) plays an important role in the progression of hypertension progress. Given hypertension is accompanied with inflammation, we, thus, hypothesized that RAS potentiates brain immune responses. Using a mouse‐derived microglial cell line, we found that prorenin caused a significant increase in TNFa expression. However, the effect of Ang II alone was minor. Interestingly, priming N‐9 cells with Ang II for 12 hr followed by LPS stimulation resulted in a dramatic increase in TNFa expression compared with the cells treated with LPS along. To determine whether microglia derived from Ang II‐hypertensive mice are similarly primed, microglia were dissociated from age matched naïve mice and the mice infused with Ang II for 2 wk. It showed that there were similar TNFa‐positive microglia populations between these two groups at base line; however, Ang II‐infused group were potentiated in that with 6hr LPS treatment, there were more TNFa‐expressing microglia compared to the control group in response to LPS. In addition there were 27% more infiltrating immune inflammatory cells in Ang II‐infused than naïve brains. These data suggest that RAS components (prorenin and Ang II) potentiate microglial reaction to inflammatory stimulators. Moreover there were increased infiltrating immune cells (inflammation) in the brain during hypertension. These findings indicate that RAS is actively involved in hypertension‐induced neuronal inflammation.
Importance of the medial amygdala to the neurogenic hypertension in Schlager mice
Authors: Jackson Kristy Larissa , Nguyen‐Huu Thu‐Phuc , Palma‐Rigo Kesia , Davern Pamela J , Head Geoffrey A
Published: June 2021

Schlager BPH mice are hypertensive due to an overactive sympathetic nervous system (SNS) and that is associated with over activity of the medial amygdala (MeAm). To determine whether the MeAm is responsible for the hypertension, we assessed the effect of MeAm ablation on blood pressure (BP) and stress reactivity. Radio‐telemetry devices were implanted in 10 normotensive (BPN) and 8 BPH mice and the MeAm was lesioned with bilateral ibotenic acid injections. One week before and 3 weeks after lesion surgery, BP was measured for 48hrs and restraint stress performed. During control measurements, BP was 121±4mmHg in BPH versus 101±2mmHg in BPN mice (P<0.001). MeAm lesions reduced 24hr BP, −11±5mmHg (P = 0.003) in BPH mice and had no effect in BPN mice. The lesion effect was similar during both day and night, suggesting a contribution independent of the state of arousal. Despite reductions in BP, the pressor response to restraint stress was well maintained. Spectral analysis revealed a reduction in low frequency mean arterial pressure (MAP) power after MeAm lesions in BPH/2J (P<0.05) suggesting that the lesions reduced SNS activity. These results show that tonic activation of the MeAm contributes at least 50% of the hypertension in BPH mice through pathways that involve activation of the SNS but which are independent of those regulating the state of arousal or the cardiovascular response to stress. Supported by NHMRC Australia
Altered regulation of sympathetic nerve activity by the caudal ventrolateral medulla in rats after exposure to chronic intermittent hypoxia
Authors: Huber Domitila Augusta , Schreihofer Ann M
Published: June 2021

After exposure to CIH, rats have elevated sympathetic nerve activity (SNA) and arterial pressure (AP). In addition, we have observed that rats exposed to CIH have reduced GABAergic inhibition of rostral ventrolateral medulla (RVLM), the source of drive for SNA. The caudal ventrolateral medulla (CVLM) is the major source of tonic inhibitory input to the RVLM. Here, we determined whether exposure to CIH reduces tonic CVLM‐mediated inhibition of SNA. Rats (8 wks old) were exposed to CIH (6% O2 for 40s, every 9 min, 8h/day, 2 wks). Then rats were ventilated, paralyzed, and prepared to record AP and splanchnic SNA. Under chloralose anesthesia rats subjected to CIH (n=9) vs. controls (n=7) had higher SNA (1.9 ± 0.2 vs. 0.9 ± 0.2 μV, P<0.05). Stimulation of CVLM by microinjection of glutamate (1 nmol/50 nl) virtually eliminated SNA (93 ± 3 vs. 94 ± 3 %) and produced bigger decreases in mean AP in CIH‐exposed rats (−39 ± 4 vs. −21 ± 3 mmHg, P<0.05). In contrast, inhibition of glutamatergic inputs to CVLM (kynurenate; 5.4 nmol/50 nl) evoked smaller rises in SNA in CIH rats (n=9) vs. control rats (n=7; 361 ± 63 vs. 605 ± 84 %, P < 0.05). These results suggest that the higher SNA in rats subjected to CIH can be eliminated by activation of the CVLM, likely via inhibition of an elevated drive from the RVLM. Furthermore, a reduced tonic inhibitory influence from the CVLM may contribute to the elevated SNA observed in rats after CIH. Support: NIHHL075174.
Interactions of Hypoxia inducible factor (HIF)‐1α and Peroxisome proliferator‐activated receptor (PPAR) α in the protection against ischemia/reperfusion injury in the rat kidney
Authors: Dallatu Mohammad , Oyekan Adebayo
Published: June 2021

In response to reduced oxygen availability, important cellular and molecular adaptive changes occur in living organisms. Hypoxia‐inducible factor (HIF) is a major adaptive mechanism and prolyl hydroxylase‐binding domain proteins (PHD) is its most important negative regulator. However, peroxisome proliferator‐activated receptor (PPARα), a nuclear transcription factor is activated in response to hypoxia/HIF‐1α and regulates many of the downstream target genes. This study tested the hypothesis that inhibition of prolyl hydroxylase domain (PHD) would confer greater protection against ischemia by a mechanism involving PPARα activation. Rats were subjected to ischemia/reperfusion (I/R) for 48 hours with or without L‐mimosine (L), a PHD inhibitor, fenofibrate, a PPARα ligand, MK886, a PPARα inhbitior. Renal blood flow (RBF; laser doppler flowmetry) recovered to 93±8%, 56±7% (p<0.001) and 87±10% (p<0.001) in control, ischemic and L‐treated groups, respectively. Coincident with these changes, serum uric acid decreased by 41±6% (p<0.05) in I/R group versus 26±3% (p<0.05) following L‐ treatment. Compared to controls, serum nitric oxide decreased by 33±3% (p<0.05) in ischemic group while L treatment abolished the effect of ischemia (P<0.05). Similarly, 24hr urine volume increased by 100±13% (p < 0.001) in I/R and this was abolished by L (3%±0). 24‐hour urinary protein excretion increased 5‐fold in I/R group but was attenuated to a 2‐fold increase in L‐treated (p<0.05) group. Fenofibrate protected rats from the effects of I/R while MK886 blunted its effect. These findings suggest that stabilization of HIF‐1α following inhibition of PHD protects against I/R in the rat kidney and that PPARα contributes to these effects.
Attenuating effect of Heme Oxygenase induction by Stannous Chloride on salt‐induced hypertension in male Sprague‐Dawley rats
Authors: Oloyo Ahmed Kolade , Ochie Daniela Ogochukwu , Bello Nurudeen Adebowale , Elias Simiat Olanike , Sofola Olusoga Adekunle
Published: June 2021

Heme oxygenase (HO) is implicated in the regulation of blood pressure both under normal and pathological conditions. Impaired vascular function leading to heightened vascular resistance is one of the mechanisms by which a high salt diet (HSD) elevates blood pressure. This study evaluated the effect of heme oxygenase induction by stannous chloride (50mg/kg body weight s.c for 4 days) on the response of BP to vasoactive substances in male Sprague‐Dawley rats fed a normal salt or high salt (0.3% and 8% NaCl) diet respectively for 6 weeks. Rats fed a HSD showed increases in arterial blood pressure (BP) [123 ± 5 vs. 102 ± 4], fluid balance (FB) [12.8 ± 1.23 vs. 6.78 ± 1.02], that was accompanied by decreased plasma concentration of bilirubin [0.84 ± 0. 11 vs. 1.36 ± 0.14] and nitrite [0.32 ±0.03 vs. 0.86 ± 0.04]. HO induction reduced the increment in BP [107 ± 5 vs. 123 ± 5] and FB [5.74 ± 0.98 vs. 12.8 ± 1.23] while restoring both bilirubin [1.30 ± 0.12 vs. 0.84 ± 0.11] and nitrite [0.70 ± 0.06 vs. 0.32 ± 0.03] level towards that of control. Likewise HO induction attenuated the blunted change in MABP in response to cumulative i.v PE and ACh that was observed in rats fed a HSD. However, change in MABP in response to i.v SNP was similar across the groups. Induction of HO via s.c stannous chloride administration attenuates blood pressure elevating effect of a HSD by mechanisms involving maintenance of FB and interactions between bilirubin and NO pathways.
Endurance exercise alters the myocardial Na+/H+ exchanger‐1 (NHE)
Authors: Starnes Joseph W , Feger Bryan J
Published: June 2021

NHE and its upregulation by hydrogen peroxide (H2O2) play a major role in myocardial pathologies. Exercise is known to improve myocardial performance and provide cardioprotection. This is the first study to examine the effect of exercise on myocardial NHE activity and its regulation by H2O2 via ERK1/2 signaling. Female S‐D rats were divided into two groups: sedentary (S, n=15) and exercised on a treadmill (E, n=11) for 5 wks. The exercise program increased (P<0.05) heart wt:body wt by 6.8% and plantaris mitochondria content by 89%. NHE activity was determined in isolated cardiomyocytes using the pH fluoroprobe BCECF. Exercise increased intrinsic NHE activity by 161% (0.57 ± 0.10 v 1.49 ± 0.18 fmol/min), which was attributed to greater cell volume (22.2 ± 0.6 v 34.3 ± 1.1 pL) and buffering capacity (28.8 ± 0.7 v 42.7 ± 0.8 mM/pH unit) in E myocytes. H2O2 stimulation (100 μM) raised NHE activity 155% in S, but only 64% in E (P<0.05). ERK1/2 inhibition abrogated the increase. Western blotting revealed that E had less total ERK1/2 content (P<0.05), but the amount of activated ERK1/2 following H2O2 was similar. No differences were observed in content of NHE or activities of the H2O2 scavengers, catalase and glutathione peroxidase. We conclude that endurance exercise training increases NHE activity at physiological pH which may enhance performance, and attenuates H2O2 stimulation of NHE activity which may contribute to protection.
Long‐term wheel running improves cardiac function but has negative consequences for diaphragmatic function in the mdx mouse
Authors: Selsby Joshua T. , Acosta Pedro , Sleeper Meg M. , Barton Elisabeth R. , Lee Sweeney H.
Published: June 2021

Dystrophin‐deficient muscles suffer from free radical injury, mitochondrial dysfunction, apoptosis, and inflammation, among other pathologies, which contribute to muscle fiber injury. Endurance exercise has the capacity to counter many of these factors. Indeed, previous work indicates that endurance training decreased disease severity in dystrophic limb muscle. Our purpose was to determine the extent to which long‐term volitional wheel running (VWR) would affect dystrophic diaphragm and cardiac function. This is especially important as the diaphragm most closely recapitulates the human disease. One year of VWR increased cardiac mass by 15% (p<0.05), left ventricle chamber size by 20% (diastole) and 18% (systole) (p<0.05), and stroke volume by 2‐fold (p<0.05) compared to sedentary mdx mice. VWR increased soleus mass by 20% (p<0.05) and tetanic force by 36% (p<0.05) while specific tension was similar between VWR and sedentary mdx mice. Notably, diaphragm specific tension was reduced by 60% (p<0.05) following one year of VWR compared to sedentary mdx mice. These data suggest that the dystrophic heart undergoes beneficial exercise‐induced remodeling. Importantly, however, the severe negative impact on diaphragm function indicates that serious caution should be taken when prescribing exercise to DMD patients. Supported by NIH grants F32AR055005–01 (JTS) and U54AR052646–03 (HLS).
Exercise prevents Western‐diet associated erectile and coronary artery endothelial dysfunction mediated by NADPH oxidase
Authors: La Favor Justin D , Anderson Ethan J , Dawkins Jillian T , Hickner Robert C , Wingard Christopher J
Published: June 2021

The purpose of this study was to investigate NADPH oxidase as a common mechanism underlying both erectile dysfunction (ED) and coronary artery disease (CAD) in inactivity and diet‐induced obesity. Male Sprague‐Dawley rats were fed a Western diet (WD) or Control diet (CD) for 12 weeks. Subgroups within each diet remained sedentary (Sed) or engaged in treadmill exercise (Ex) throughout the dietary intervention. Erectile function was evaluated by measuring the mean arterial pressure and intracavernosal pressure in response to electrical field stimulation of the cavernosal nerve, prior to and following an intracavernosal injection of apocynin (Apo; an NADPH oxidase inhibitor). Coronary artery endothelial function (CAEF) was evaluated ex vivo with cumulative doses of acetylcholine (ACh) applied to coronary artery segments pre‐constricted with serotonin. CAEF was assessed in the presence and absence of Apo. Erectile function (p < 0.0001) and CAEF (p < 0.001) were attenuated in WD‐Sed rats. Erectile function (p < 0.01) and CAEF (p < 0.05) were augmented by Apo only in WD‐Sed rats. Ex preserved erectile function (p < 0.0001) and CAEF (p < 0.05) within the WD. This study suggests that erectile function and CAEF are impaired by NADPH oxidase in chronic WD‐fed rats, while Ex training may be a practical means of preventing WD‐induced ED and CAD.

Supported by the Sexual Medicine Society of North America.
Effects of ASEA beverage intake on endurance performance in mice
Authors: Knab Amy M , Nieman David C , Shanely R. Andrew , Zwetsloot Jennifer J , Cialdella‐Kam Lynn , Meaney Mary Pat
Published: June 2021

Previous research with ASEA (a saline beverage with stable superoxide complexes) found increased fatty acid mobilization in cyclists after 7 d ingestion. We hypothesized that run time to exhaustion would be favorably influenced by ASEA intake due to enhanced fatty acid oxidation and muscle glycogen sparing. Sixty mice were randomized to 1 of 4 treatment groups (n=15 each): placebo sedentary (PS), ASEA sedentary (AS), placebo run (PR), and ASEA run (AR). Mice were gavaged daily with ASEA or placebo (0.3 ml/d) for 7 d. PR and AR groups were run to exhaustion (24 m/min) at the end of the 7‐d gavage period, with AR running significantly longer than PR (68.0±9.2 vs. 52.8±7.4 min, respectively) (p<0.001). At the point of exhaustion, liver glycogen was undetectable for both AR and PR. When adjusted to run time, the estimated rate of muscle glycogen depletion was different between AR and PR (0.036±0.014 and 0.052±0.018 ug/mg protein per min, respectively, p=0.017). Skeletal muscle phosphorylated acetyl‐CoA carboxylase (p‐ACC) was significantly increased in AR compared to AS (p=0.020) and PR (p=0.045). Fatty acyl CoA transport (CPT1), and beta‐oxidation (beta‐HAD) were not different between AR and PR. ASEA increased run time to exhaustion by 29% in mice, potentially through less inhibition of fatty acid oxidation via increased P‐ACC, and muscle glycogen sparing percent (30%).

Funded by Reoxcyn Discoveries Group.
Kinetic examination of femoral bone development in modern broiler chickens
Authors: Prisby Rhonda D , Campbell Jeremiah , Menezes Thomas , Benson Tyler , Samraj Enoch , Pevzner Igal Y. , Wideman Robert
Published: June 2021

BACKGROUD

Lameness in broiler chickens often presents as femoral head separation, degeneration and necrosis. The etiologies of these disorders are unknown and may results from compromised blood flow to femora and/or imbalances in bone resorption and formation.

METHODS

We examined developmental changes in femoral bone volume and metabolism, via histomorphometry, at 7, 15, 21, 28, and 42 days. Bone volume (BV/TV, %), osteoid surface (OS/BS, %), osteoblast activity (Ob.S/BS, %) and osteoclast activity (Oc.S/BS) were evaluated.

RESULTS

Body mass increased 23‐fold from 7 to 42 days. BV/TV remained stagnate (range: 5%–8%) and reached its highest value at day 28. Bone volume‐to‐body mass ratio (range: 0.057%–0.002%), Ob.S/BS (range: 1.3%–0.5%) and OS/BS (range: 22.9%–5.6%) declined significantly (p < 0.05) with age and Oc.S/BS was so excessive throughout development that analysis was not possible. In addition, vast areas of chondrocyte columns often expanded into the secondary spongiosa.

CONCLUSIONS

Development of broiler femora failed to sufficiently match overall body growth. The lack of bone development corresponded to diminished osteoblast and augmented osteoclast activity. The large chondrocyte columns possibly coincide with a lack of blood vessel penetration that is necessary for calcification of the cartilage template and subsequent bone development. Supported by Cobb‐Vantress Inc.
Inactivity Suppresses the Expression of Amino Acid Transporters in Rat Skeletal Muscle
Authors: Murakami Taro , Ishiguro Nahoko , Yoshinaga Mariko
Published: June 2021

Leucine is one of the building blocks of protein but also stimulates the translation in protein synthesis through the activation of mTORC1 signaling. Furthermore, leucine is oxidized as a fuel to synthesize ATP during exercise in the muscle. These features of leucine let us hypothesized that the expression of leucine transporters would be modified in proportion to its cellular demand induced by physical activity. This study investigated whether the expression of leucine transporters would be induced by voluntary running and reduced by inactivity following the exercise in rat skeletal muscle. Thirty male Fisher 344 × Brown Norway F1 hybrid rats were obtained at 21 days of age. Rats were separated into those with access to running wheels (n=15, wheel lock, WL) and those without access to the wheels (n=15, SED). Forty days after the beginning, wheels were locked at 06:00 and rats were killed 5, 53 or 173 h after locking of wheels. LAT1, CD98, SNAT2, and PAT1 mRNAs in soleus, gastrocnemius, and EDL muscles were measured by qRT‐PCR. Soleus muscle weight was increased by voluntary running and reduced by inactivity following voluntary running. The expression of mRNAs for LAT1 and PAT1 was also induced by voluntary running and reduced by inactivity following it. These results suggested the expression of leucine transporters would be involved in a muscle hypertrophy and atrophy induced by physical activity and inactivity, respectively.
INTRAUTERINE GROWTH RESTRICTION MODIFIES THE NORMAL GENE EXPRESSION IN KIDNEY FROM RABBIT FETUSES
Authors: Lozano Mauricio , Figueroa Horacio , Suazo Cristian , Illanes Sebastian , Eixarch Elisenda , Gratacós Eduard , Hernández‐Andrade Edgar , Irarrazabal Carlos E
Published: June 2021

Intrauterine growth restriction (IUGR) is associated with increased risk of neonatal complications; birth hypoxia; impaired neuronal and kidney development; and stillbirth.

New Zealand white pregnant rabbits were used to establish a model of IUGR. Rabbit fetuses with twenty‐five days of gestation (25D) were subjected to severe restriction by 40–50% ligature of uteroplacental vessels. At day 30 (120 hrs after surgery) newborns were obtained by caesarean. After delivery, living newborns were weighed. Kidneys were extracted from 39 fetal rabbits (19 controls and 20 IUGR) and gene expression of genes related with hypertonicity (NFAT5), hypoxia (HIF‐1α), and endothelial activity (Arg2, iNOS and eNOS) were evaluated by qRT‐PCR.

Experimental IUGR fetuses were significant smaller than controls (39 vs. 48gr, p<0.05). The qRT‐PCR analysis showed a significant increasing in the NFAT5 (1.7 fold, p<0.05), iNOS (3.4 fold, p<0.05) and Arg2 (6.3 fold, p<0.05) mRNA. On the other hand, a significant reduction of eNOS mRNA level (0.34 fold, p<0.05) was observed. The HIF‐1α did not show changes in its mRNA expression.

The selective ligature of uteroplacental vessels in the pregnant rabbit demonstrated an important mRNA dysregulation of essential proteins involved in fetal kidney development and adult kidney maintenance. Fondecyt 1100885 and Fondecyt 1110869.
Isocapnic augmented exercise ventilation in congestive heart failure: effect of apparent (real‐feel) metabolic CO2 load in respiratory chemosensing
Authors: Poon Chi‐Sang
Published: June 2021

CO2 breathing introduces an airway CO2 load that clogs the mechanism of pulmonary CO2 elimination, inevitably causing hypercapnia when the inspired PCO2 is high enough to prohibit isocapnia regardless of the ventilation level. The control law (Comroe's law) evidencing such self‐imposed permissive hypercapnia has been dubbed “CO2 tolerance curve” (rather than CO2 response curve) by Fenn and Craig (1963). By contrast, muscular exercise introduces a metabolic CO2 load that is readily countered with proportionate increase in ventilation without CO2 retention. Isocapnia from rest to exercise is maintained by the controller with the exercise ventilatory response being well adjusted for corresponding decrease in anatomical VD/VT, resulting in a small positive Y‐intercept in the ventilation vs. metabolic CO2 output relationship (Whipp's law). Remarkably, patients with congestive heart failure compensate for their abnormally large alveolar VD/VT with augmented ventilation, maintaining isocapnia at rest and during exercise without CO2 retention despite their decreased exercise tolerance. Thus, the control law governing isocapnic exercise ventilatory response is not merely determined by metabolic CO2 output per se but is responsive to an apparent (real‐feel) metabolic CO2 load that also accounts for the adverse effect of physiological VD/VT on pulmonary CO2 elimination. (Supported by HL067966 and RR028241)
Cerebral erythropoietin regulates the respiratory control system during early postnatal life
Authors: Caravagna Céline , Schneider Gasser Edith M. , Soliz Jorge
Published: June 2021

Apart from its role in neuroprotection, erythropoietin (Epo) synthesized by neurons and astrocytes modulates the respiratory control system in adult mice. However, the impact of Epo in the regulation of the neural control of breathing during the postnatal development remains unknown. Here we use a transgenic mouse line (Tg21) constitutively overexpressing human Epo in brain to investigate whether Epo interacts with the brainstem centers associated with respiration during postnatal life. Our immunohistochemical analysis revealed that Epo and Epo receptor (EpoR) are expressed in the main central respiratory nuclei at 1, 4, 7, 10, 15 and 21 days of age. Furthermore, by using antibodies to choline acetyltranferase (ChAT), somatostatin (SST) and neurokinin receptor 1 (NK1), EpoR was localized to the pre‐Bötzinger complex nucleus at all studied postnatal days. Whole body phtethysmography technique was used to evaluate basal ventilation and ventilatory response to hypoxia at 10% and 6% O2. Compared to wild type animals, Tg21 mice showed improved hypoxic ventilation (both at 10% and 6% O2) at postnatal days 15 and 21, but not 7 and 10. These results are in line with data obtained by radioimmunoassay, showing that overexpression of Epo in transgenic mice brains dramatically increases after 10 days of postnatal life. These results imply that cerebral Epo plays a key‐regulating role in the neural control of breathing in a period in which the central respiratory network undergoes full development and maturation.

Source of research support is Molly Towell Perinatal Research Foundation (MTPRF).
Prolonged intermittent hypoxia activates p38 MAP kinase in the phrenic motor nucleus
Authors: Peterson Timothy J , Huxtable Adrianne G , Mitchell Gordon S
Published: June 2021

Plasticity is a fundamental property of the neural system controlling breathing. Although inflammation is prominent during most clinical disorders that challenge ventilatory control, little is known concerning the influence of inflammation on respiratory plasticity. One trigger to systemic (and neural) inflammation is prolonged (one day) intermittent hypoxia (IH‐1; 2 min hypoxia/2 min normoxia, 8 hrs), simulating intermittent hypoxia during sleep disordered breathing. IH‐1 abolishes an important model of respiratory plasticity, phrenic long‐term facilitation, but this effect is reversed by spinal application of a p38 MAP kinase inhibitor. p38 MAPK both initiates and is activated by inflammation. Here, we sought to determine which cells in the phrenic motor nucleus express increased phosphorylated (activated) p38 MAPK following IH‐1. Using immunofluorescence (n=5), increased phospho‐p38 MAPK was detected in identified phrenic motoneurons (Cholera Toxin B labeled) and adjacent microglia (cd11b positive cells). These results suggest that p38 MAPK activity may impair pLTF following IH‐1 by: 1) disrupting pLTF within phrenic motor neurons; and/or 2) initiating microglial inflammatory cascades that indirectly impair pLTF. p38 MAPK may be a viable therapeutic target to reduce the impact of inflammation on respiratory plasticity.

Supported by HL111598, APS UGSRF Program
Functional glycomics of the pulmonary and systemic vasculature: Lectin histochemistry for probing differentiated glycoproteins
Authors: Cioffi Eugene A , Finnorn Brandon , Cioffi Donna L
Published: June 2021

Endothelial cells line the inside of blood vessels and form a semi‐permeable barrier. Curiously, arteriosclerotic vascular disease (ASVD) is frequently exhibited within the systemic vasculature, whilst ASVD is rarely observed within the pulmonary vasculature. Since both vessel types transport the same components, this suggests some differences between the pulmonary and systemic endothelium that may underlie development of ASVD. We hypothesize that the surficial glycocalyx of the vessels plays a significant role in arteriosclerotic plaque formation.

The glycan compositions of the pulmonary and systemic vasculatures have not been examined in detail. We compared rat pulmonary artery (PAECs) and rat pulmonary microvascular endothelial cells (PMVECs) with rat aortic endothelial cells (AOECs, systemic) and human coronary artery cells (HCACs, systemic) using a panel of fluorescently‐tagged lectins; key differences in the surficial carbohydrate expression on the glycocalyx in the pulmonary versus systemic vasculatures was observed. Most notably, systemic AOECs and HCACs showed a very high expression of 9‐acetylated sialic acid (9‐OAc‐Neu5Ac), whilst the pulmonary PAECs and PMVECs had little. In contrast, the pulmonary cells strongly expressed terminal á‐mannose, whereas the systemic cells showed a weak á‐mannose expression. Supported by 5R00HL089361.
Leptin and adiponectin have opposite effect on ciliary activity, calcium wave velocity and ovum transport velocity in the rat oviduct
Authors: Oses Carolina , Milovic Carlos , Llados Carmen , Tejos Cristian , Villalón Manuel
Published: June 2021

Obesity (with high leptin and reduced adiponectin serum concentrations) correlates with reduced fertility. Embryos oviductal transport depends on coordinated ciliary activity and smooth muscle contractions, which are modulated by intracellular calcium levels. We hypothesize that leptin and adiponectin affect ciliary beat frequency and intracellular calcium homeostasis, changing the embryos transport velocity through the rat oviduct. Effect of different leptin or adiponectin concentrations (1, 10 and 100 ng/ml) over ciliary beat frecuency was tested on primary cultures of ciliated cells from the rat epithelium. We observe a transient decrease in ciliary beat frequency with leptin and increase with adiponectin. High leptin levels resulted in a reduction on propagation velocity of intracellular calcium wave produced after a mechanical stimulation in 24 hrs treated cell cultures, and adiponectin showed the opposite effect. We test in vivo effects in female rats, on day 1 of pregnancy, leptin or adiponectin (100 ng/ml) were injected in one side of ovarian bursa and vehicle in the contralateral side. On pregnancy day 5, leptin induces retention of the embryos in the oviduct, in other wise, adiponectin have no effects on embryonic transport all of the embryos were finding in uterus. These provide a possible explanation for the cellular effect of obesity on the reduced female fertility (ACT‐79 and CONICYT 24121342).
MicroRNA‐29 expression in pancreatic islets correlates with the impairment of maternal glucose homeostasis induced by glucocorticoid excess in late pregnancy
Authors: Gomes Patricia Rodrigues Lourenço , Graciano Maria Fernanda Rodrigues , Pantaleão Lucas Carminati , Lellis‐Santos Camilo , Carpinelli Angelo Rafael , Latorraca Marcia Queiroz , Bordin Silvana
Published: June 2021

We investigated the long‐term effects of dexamethasone (DEX) administration during late gestation in glucose homeostasis and in the expression of miRNAs in pancreatic islets. Pregnant Wistar rats received dexamethasone (DEX), 0.2 mg/kg daily in drinking water during the last five days of pregnancy (Preg‐DEX). Age‐matched virgin female rats either treated (CTL‐DEX) or not (CTL) with DEX, as well as untreated pregnant rats (Preg) were used as controls. All groups were subjected to glucose and insulin tolerance tests at weaning and at 3, 6, and 12 month after weaning. Pancreatic islets were isolated for glucose stimulated insulin secretion (GSIS) and miRNA expression. Compared to control groups, glucose intolerance was observed in Preg‐DEX rats from weaning to 12 months after weaning. In all periods insulin sensitivity was normal. An upregulation in miR‐29 family (miR‐29a/b/c) was observed in Preg‐DEX islets at the end of gestation and 12 months after weaning. In the later period, mir‐29 expression paralleled a decrease in GSIS. In conclusion, we show that the excess of glucocorticoid in late pregnancy permanently impairs pancreatic islets function, which is correlated with persistent upregulation of miR‐29 expression.

Financial support: FAPESP 2011/01568–5, PRONEX‐FAPEMAT.
The “Luck‐Free” Exam: Promoting Transparency, Encouraging Collaboration and Active Learning
Authors: Rangachari P K , Nastos Stash
Published: June 2021

Examinations can be powerful stimuli to collaborative learning, but are rarely used as such, since they can be stressful. We attempted to make a formal exam a good learning experience for students in a large undergraduate freshman biology course (average class size 175). To defuse anxiety and reduce the element of luck, students were given a set of 8–10 questions, well in advance of the exam. These questions probed their understanding of the material taught, and required them to seek, synthesize and integrate information from diverse sources. We encouraged them to collaborate in groups to frame suitable answers, and solidify what they had learned within the class setting. The students knew that the final formal exam would be an individual one, where they would get a smaller subset of the very same questions. Their answers clearly showed that they had understood the core concepts of the course. Over a 4‐year period, 612 students rated the value of this assessment to their learning experience, on a 10‐point scale: median 8, mode 10, range 1–10. The students appreciated the opportunity to solidify their learning in this fashion, and rated their learning experience highly. We thank the Canadian taxpayers for still supporting public universities.
Developing online basic nutrition curriculum based on an existing face‐to‐face course
Authors: Stewart Maria L.
Published: June 2021

FSHN 185: The Science of Human Nutrition is an introductory nutrition course required of many degree programs at the University of Hawaii at Manoa (UHM). An online version of this course was developed with assistance from the Digital Course Design and Consulting Group at UHM College of Education during Spring 2011. The course was first offered Fall 2011. The goals of the online course were to 1) meet the same SLOs as the face‐to‐face class, 2) offer greater flexibility for students and 3) to attract students from beyond the UHM campus. The online course content is embedded in the UHM Sakai‐based course management system, Laulima, and presented as weekly modules. All assessments are conducted within Laulima. The content utilizes Blackboard Collaborate for synchronous interactions and archived lectures. Custom animations and review activities were created with Camtasia and StudyMate, respectively. Average final course grades in the online sections were comparable to the face‐to‐face sections taught by the same instructor. Student teaching evaluations were not significantly different between the online and face‐to‐face sections. Most students in the course resided on the island of Oahu. Off campus students completed the course from locations such as Commonwealth of the Northern Mariana Islands and Alaska. Partial funding for this project was provided by the College of Tropical Agriculture and Human Resources‐ UHM.
GORILLA GORILLA: AN ANATOMICAL, DISMANTABLE, REAL SIZED MODEL IN PAPIERT MACHE OF THE 19th CENTURY OF THE AUZOUX FACTORY
Authors: LE FLOCH‐PRIGENT Patrice , GILLOT Jean Bernard , BARBET J. Patrick
Published: June 2021

The anatomy of the Gorilla is poorly studied and published, most often in a partial or incomplete manner. In the second half of the19th century, AUZOUX a french physician built anatomical models in papier‐mâché which were internationally spread. He realized a real‐sized model of a female Gorilla after a dissection of its own. He chose aesthetic aspects as the grasping of branches and a muscular representation on the lefts side versus an osseous one on the left. This model is really a scientific document as it recorded the name, function and innervation of the muscles on their hidden parts. These data remains unique. This model was non colored; several known other ones were colored. The production was probably very limited.
ANALYSIS OF BCL‐2 MYOCARDIAL GENE EXPRESSION IN FETUSES OF DIABETIC CD1 MOTHERS ON DAYS 14 AND 17 OF GESTATION
Authors: Gutierrez Juan Claudio , Navarrete Patricia
Published: June 2021

The objective of this study was to demonstrate that Bcl2 gene expression was dys‐regulated in fetal myocardium from diabetic mothers on days 14 and 17 of gestation. Fifty two female CD1 mice, 6 to eight 8 weeks old were used on these experiments. After a 2 weeks acclimation period, females were divided in three experimental groups: control, diabetic (STZ) and diabetic treated with FCA (STZ + FCA). Results from RT‐PCR showed that Bcl2 expression was decreased for the STZ and STZ + FCA groups at GD14 and 17. Body weight was significantly lower at GD14 fetuses from diabetic litters than in controls. Also, at DG17, there were no significant differences among groups in this parameter. Fetal body length and number of fetuses per litter did not show significant differences among groups. External congenital malformations were found in both groups treated with STZ. At GD 17 there was a reduction of birth defects from 21% in fetuses from diabetic mothers to 15% in fetuses from diabetic mothers treated with FCA.
Physiological Assessment Of The PEPCK‐Cmus Mouse Heart. A Mouse Model Of An Athletic Heart Without Imposed Training?
Authors: Flores Saul , Hakimi Parvin , Liner Anna , Yu Xin , Hanson Richard W. , Snyder Christopher S. , Hoit Brian D. , Watanabe Michiko
Published: June 2021

Background

Cardiac hypertrophy in athletes is adaptive and distinct from pathological hypertrophy. Mice that over‐express phosphoenolpyruvate carboxykinase (PEPCK‐C) in skeletal muscle are more active and live longer compared to controls. We hypothesized that PEPCK‐Cmus mice would have an athletic cardiac phenotype in the absence of an imposed exercise regimen.

Methods

Analysis of activity level, cardiac structure with conventional (M‐mode, Doppler) and deformational (midwall strain, strain rate) echocardiographic parameters, and cardiac MRI for torsion and time to peak strain (Tpk) in PEPCK‐Cmus and control mice.

Results

The heart rate, end diastolic dimension, LV fraction shortening and ventricular torsion were similar between groups.

Measurement
Unit
Control (n=5)
PEPCK‐Cmus (n=6)

Activity

Distance moved
cm
61999 +25.2
85830 +33.5*

Echocardiography

RWT
cm
0.56 +0.13
0.89 +0.16*

LVMI
mg/weight
3 +0.11
5+0.11*

MPI (LV)

0.45 +0.02
0.35 +0.07*

CO
cc
0.09 +0.02
0.24 +0.06*

CS
%
−27.4 +5.7
−49.1+3.1*

CS rate
1/s
−7.7 +1.8
−17.5+1.9*

RS
%
+16.9 +1.2
+37.386 +5.9*

RS rate
1/s
+4.6 +1.1
+7.8 +1.6*

Cardiac MRI

Tpk
%
60 +1.2
40 +2.1*

P‐value <0.05,

RWT=relative wall thickness; LVMI=LV mass index; MPI=myocardial performance index; CO=cardiac output; CS=circumferential strain; RS=radial strain

Conclusions

Untrained PEPCK‐Cmus mice have greater activity and their hearts have phenotypical changes characteristic of the athlete's heart, and are a novel model for non‐pathological cardiac hypertrophy that may be useful for comparison with other mice models of hypertrophy.

The radial and circumferential strain measurements were distinctly different in the PEPCK‐Cmus mice indicating these parameters should be considered when clinically assessing hypertrophy.

Our future plan is to analyze molecular changes (VEGF, PECAM) by Western blots in PEPCK‐Cmus and control mice.

Grant Funding Source: N/A
Are adherens junctions implicated in the transition from physiological to pathological cardiac hypertrophy?
Authors: Prado Cibele M , Blefari Valdecir , Santos Daniele O. , Malvestio Lygia MM , Pedro Rubens F. , Rossi Marcos M.
Published: June 2021

Hypertension causes cardiac hypertrophy, cardiac dysfunction, and heart failure (HF). The mechanisms implicated in the transition from physiological to pathological hypertrophy are still unknown. Growing evidence points out to the role of inappropriate mechanical coupling in the pathogenesis of HF. We investigated adherens junctions and its potential role in the pathogenesis of HF in rats submitted to chronic abdominal aorta stenosis. Wistar rats were submitted to abdominal aorta stenosis and killed at 90 days post surgery: 60% presented hypertrophied hearts (HH) and 40% hypertrophied and dilated hearts (DH), as evaluated by echocardiography. DH and HH were compared to control hearts. The hearts were collected and Western blot (WB) and immunofluorescence (IF) were performed for cadherin, beta‐catenin and calpain. Blood pressure was evaluated. Data were considered significant when p<0.05. Cadherin reduced 20% in HH and 64% in DH; beta‐catenin reduced 32% in DH and calpain increased 50% in DH as compared to control. The ejection fraction reduced 35% in DH. Adherens junctions remodeling may be partly responsible for the development of heart failure in rats submitted to chronic abdominal aorta stenosis. These abnormal parameters may emerge as therapeutic targets to modulate the evolution of heart hypertrophy from compensated to decompensated phase. Grants from FAPESP 09/17787–8; 09/54010–1; 10/19216–5.
Capacity for the Cretaceous pterosaur Anhanguera to launch from water
Authors: Habib Michael
Published: June 2021

Many large pterosaurs come from marine sediments. While the feeding ecology of large pterosaurs is contentious, it is likely that some species took food from the water surface. As a result, it is likely that some pterosaurs were forced into the water through stochastic events. Unless a water launch was possible, such events would presumably be lethal. Here I present the results of a quantitative and comparative morphological analysis of the large marine pterosaur Anhanguera santanae that suggest this animal could launch from water, without the assistance of wind or waves, using a form of quadrupedal takeoff. Initial escape from the water surface would use the most energy and could be accomplished with a net remaining acceleration of 17 m/s2, with an acceleration of up to 39 m/s2on the subsequent series of repeated propulsive bounds, which would have occurred as a series of “hops” across the water surface. While the capacity for such a behavior is not demonstration that such it was utilized, this species possessed a wide array of anatomical features that would make water launch both efficient and effective, and cannot be easily explained as adaptations to flight or walking: the expanded scapula, reinforced scapular‐notarial joint, warped deltopectoral crest, exceptionally broad MCIV‐PHIV wing finger joint, and expanded posterior brachial musculature. Funding provided by the Jurassic Foundation.
Role of Sca‐1 after ischemia/reperfusion injury in adult mouse kidney
Authors: Camarata Troy , Vasilyev Aleksandr , Arnaout Amin
Published: June 2021

Acute kidney injury (AKI) is a common clinical problem with significant consequences on healthcare. It is of great importance to understand the genetic and cellular changes induced by AKI. Stem cell antigen‐1 (Sca‐1, Ly6a), is a GPI‐anchored protein expressed on progenitor cell as well as differentiated cell populations. Although commonly used as a marker to enrich for stem/progenitor cells, little is known about the function of Sca‐1. Recent work has suggested Sca‐1 may play role in Smad activation downstream of TGFβ receptor 1, a pathway known to function in the progression of kidney disease. We sought to determine the role of Sca‐1 in the adult kidney using ischemia/reperfusion injury (IRI) as a model for AKI. Sca‐1 is expressed in epithelial cells of the proximal tubule, loop‐of‐Henle, and distal tubule as well as in specific interstitial and endothelial cells. After IRI, Sca‐1 knockout mice displayed significantly increased kidney injury molecule 1 expression and prolonged Smad2 phosphorylation compared to wild‐type controls. Interestingly, uninjured Sca‐1 knock‐out mice also had increased Smad2 activation. In addition, loss of Sca‐1 resulted in increased cytokine and collagen expression after IRI. These results suggest Sca‐1 is required to inhibit Smad activation by the TGFβ pathway and may protect from AKI induced interstitial fibrosis.

Grant Funding Source: N/A
DNA repair choice defines a common pathway for recruitment of chromatin regulators
Authors: Bennett Gwendolyn , Papamichos‐Chronakis Manolis , Peterson Craig
Published: June 2021

DNA double‐strand break (DSB) repair is essential for maintenance of genome stability. Recent work has implicated a host of chromatin regulators in the DNA damage response, but their functional roles remain unclear. In budding yeast, DSB recruitment of chromatin regulators has been monitored primarily in asynchronous cell populations, and thus it is unclear if these events are linked to non‐homologous end joining (NHEJ) or homologous recombination (HR). By using synchronized cell populations, we surprisingly find that subunits of the INO80, SWR‐C, NuA4, SWI/SNF, and RSC enzymes, as monitored by chromatin immunoprecipitation (ChIP), are primarily recruited outside of the G1 cell phase. Furthermore, recruitment is inhibited by the NHEJ machinery, and requires early steps of HR, strongly supporting a role for these enzymes in the HR repair pathway. Strikingly, and in contrast to previous reports, we also find no significant role for H2A.X phosphorylation (γH2AX) in the recruitment of chromatin regulators, but rather their recruitment coincides with reduced levels of γH2AX.

This work was supported by a grant from the NIH (GM54096).
A unique acquired resistance mechanism to agents that target GLI (GANT61) in the Hedgehog (HH) signaling pathway
Authors: Agyeman Akwasi , Mazunder Tapati , Houghton Janet
Published: June 2021

Canonical HH signaling engages the transmembrane receptor PTCH, the intermediary signaling molecule SMO, and the HH signaling response molecules, GLI1 and GLI2. In colon cancers, GLI is aberrantly regulated, driving cell survival and genomic instability in oncogenesis, progression and metastasis. Following inhibition of GLI by a protracted exposure of HT29 to GANT61, we have identified a previously undescribed, novel mechanism by which cancer cells evade cell death. Stepwise selection did not produce a proliferative colon cancer cell line resistant to GANT61, confirming the critical importance of GLI genes to cell survival. In contrast, in HT29 cells, by inhibiting GLI1/GLI2 by prolonged exposure to semi‐lethal concentrations of GANT61 (10 μM; G10), we observed reduced proliferation, decreased appearance of γH2AX nuclear foci, and increased methylation of histone H3K9 (H3K9me1–3) within the promoters of the GLI1 and GLI2 genes. Increased H3K9 methylation was also demonstrated in HT29(G10) cells in the promoter regions of FoxM1 and CDC6, a G1/S and PreReplication Complex assembly regulatory genes, respectively. Data suggest that the GLI2‐>;GLI1‐>;FOXM1‐>;CDC6 promoters are coordinately regulated by H3K9 methylation in a pathway to promote gene silencing, reduce gene transcription, and reduce cell cycle transition at G1/S, constituting a unique mechanism of acquired resistance to drugs that target GLI.

This work is supported by NCI awards RO1 CA 32613 (J.A. Houghton) and NCI RO1 108929 (J.A. Houghton)