bluuebunny/crossref_metadata_2025 huggingface dataset:
Activation of NLRP3 inflammasome in alveolar macrophages mediates mechanical stretch‐induced lung inflammation
Authors: Wu Jianbo , Yan Zhibo , Malik Asrar , Schwartz David , Yu Jingui , Hu Guochang
Published: June 2021

Mechanical ventilation is capable of activating the innate immune system and inducing sterile inflammatory response. The proinflammatory cytokine IL‐1β is among the definitive markers for accurately identifying ventilator‐induced lung inflammation. However, mechanisms of IL‐1β release from inflammatory cells during mechanical ventilation are unknown. Here we show that cyclic stretch activates the NLRP3 inflammasomes and induces release of IL‐1β and IL‐18 in mouse alveolar macrophages via caspase‐1‐ and TLR4‐dependent mechanisms. We also observed that gp91phox reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit is dispensable for stretch‐induced cytokine production whereas mitochondrial generation of reactive oxygen species (ROS) was required for stretch‐induced NLRP3 inflammasome activation and IL‐1β release. Furthermore, mechanical ventilation with a high tidal volume activates NLRP3 inflammasome in mouse alveolar macrophages and increases the production of IL‐1β and IL‐18 in vivo. These findings suggest that alveolar macrophage‐associated NLRP3 inflammasome has an airway stretch‐sensing capacity that induces the release of IL‐1β, and hence may contribute to the mechanism of lung inflammatory injury.
In vitro AAV5‐mediated expression of metalloendopeptidase neurolysin in mouse brain primary cultures
Authors: Wangler Naomi J. , Sumners Colin , Speth Robert C. , Karamyan Vardan T.
Published: June 2021

The aim of this study was to characterize a newly developed adeno‐associated virus serotype 5 (AAV5) vector encoding mouse neurolysin in primary cultures of neurons and astroglia from mouse brains. Cortical neurons and astroglia were prepared from E16 embryos and 1‐day‐old CD‐1 mice, respectively. Neurons were transduced on in vitro day (ivd) 7, and astroglial cells on ivd 21 using AAV5‐neurolysin or control AAV5‐eGFP vectors with chicken β‐actin promoter at a concentration of 1.8 × 1011 genome copies per 100 mm dish. Activity of neurolysin (sensitive to inhibition by 10 mM Pro‐Ile) in cytosolic and membrane fractions was determined two weeks after transduction by cleavage of a specific quenched fluorogenic substrate. Relative quantity of neurolysin in the same samples was determined by Western Blotting using a specific antibody. In neuronal membranes, neurolysin activity and protein were significantly upregulated: 50% increase in activity, and 40% increase in density. Cytoplasmic increases in neurolysin activity and protein were 198% and 248%, respectively. In contrast, neurolysin activity and protein amount in astroglial membranes and cytoplasm showed a marginal, non‐significant increase. These results demonstrate the ability of newly developed vector to transduce functional neurolysin in neurons making it an excellent tool to study (patho)physiological function of this peptidase.
Characterization of Axon Innervation to the Extraocular Muscles in Wild Type Mice
Authors: Garcia Ariel Elizabeth , Nugent Alicia , Engle Elizabeth
Published: June 2021

Certain forms of human strabismus result from abnormal ocular cranial nerve development, such that the nerves do not reach appropriate eye muscle targets, causing stereotyped defects in eye movement. Human eye movement disorders are now being modeled in mice. Little is known about the three‐dimensional innervation pattern of ocular cranial nerves onto extraocular muscles during mouse development. This research aims to gain an understanding of the normal developmental growth and innervation of ocular cranial nerves III, IV, and VI from their brainstem nuclei to the extraocular muscles in wild type mice to form a database from which to compare innervation patterns of normal and mutant mouse models. ISLMN:GFP reporter mice on a mixed background were used to visualize ocular cranial nerve input onto immunostained extraocular muscles during embryonic development. Histological sections were reconstructed into 3D images at multiple embryonic time points between E11.5 and E17.5 to determine innervation patterns. Characterization of normal extraocular muscle innervation during mouse development will facilitate the identification of innervation defects in mouse models of human eye movement disorders.
Angiotensin II change glutamate uptake and GFAP expression in astrocyte culture from hypothalamus
Authors: Cruz Josiane Campos , Lopes Milene Mantovani , Elias Lucila Leico Kagohara , Antunes‐Rodrigues José
Published: June 2021

Central ANG‐II induce vasopressin and oxytocin release (Reis et al. Behav Brain Funct. 2010). Considering that dehydration diminishes glutamate uptake by hypothalamic astrocyte, as well induce retraction of astroglial processes (Boudaba et al. J Physiol 2003). We hypothesized that ANG II could modulate magnocellular excitability through a decrease of glutamate uptake and/or change in glial fibrillary acidic protein (GFAP) expression in hypothalamic astrocytes. We aimed to study the role of ANG II modulating glutamate uptake and GFAP expression in hypothalamic astrocytes cultured. Astrocyte culture were stimulated with ANG‐II (1, 10 and 100 nM) during 5, 15 and 30 min. Glutamate uptake was evaluated through [3H] aspartate assay and GFAP expression was evaluated after ANG‐II (100 nM) stimulus during 5, 15 and 30 min through Western Blot Analysis [protein membrane were incubated with mouse α‐GFAP (1:500) or β‐galactosidase (1:3000)]. The results showed that ANG‐II time‐and concentration‐ dependent induced a significant decrease in aspartate uptake. Maximal decrease was observed at 5 min with 100 nM Ang II [47±5.2% under control (n= 11)]. ANG‐II induced a time‐dependent increase in the GFAP expression, with maximal increase at 15 min [57±24% above control (n=06)]. The results suggest that the diminished glutamate uptake induce by ANG‐II increases the availability of extracellular glutamate within the hypothalamus, which could contribute to an increased magnocellular neuronal excitability. In addition, GFAP expression induced by ANG‐II could act modulating availability of extracellular glutamate. Financial Support: FAPESP, CNPq.
Prevertebral sympathetic ganglion neurons projecting to mesenteric veins are less responsive to exogenous glucocorticoid in DOCA‐salt hypertension
Authors: Shah Amit , Houchin Timothy , Kreulen David
Published: June 2021

The superior mesenteric ganglion (SMG) contains sympathetic postganglionic neurons that innervate the blood vessels of the small intestine. In hypertension (HT), capacitance in veins may be reduced, resulting in a shift in blood volume to less compliant arteries. We investigated the properties of neurons of the SMG that project to mesenteric arteries or veins and compared these properties in intact ganglia from adult normotensive (NT) and DOCA‐salt HT rats. We inoculated segments of mesenteric veins and arteries with pseudorabies‐viral constructs that resulted in expression of fluorescent proteins in the somata. Intracellular recordings of identified vein and artery neurons revealed resting membrane potentials of −43 and −47 mV (n=19 and 13), cell input resistances of 63 and 76 MΩ, and membrane time constants of 12 and 13 ms. These properties were not significantly different between NT and HT rats (n=98 neurons total). Application of the glucocorticoid, hydrocortisone 21‐hemisuccinate (100 μM), resulted in a 5mV hyperpolarization (n=7) in artery neurons from NT and HT rats, and in vein neurons from NT rats. When applied during recording of vein neurons from HT rats, however, the amplitude of this hyperpolarization was reduced to 1mV (n=9) P=0.0005. This result suggests that neurons projecting to mesenteric veins are differentially modulated in DOCA‐salt HT.
Central Histaminergic System Mediates Prostaglandin Cascade Induced Cardiovascular Effects
Authors: Yalcin Murat , Altinbas Burcin , Topuz Bora Burak , Yilmaz Mustafa Sertac , Savci Vahide , Aydin Sami
Published: June 2021

Previously, we showed that central injection of melittin, a phospholipase A2 activator, arachidonic acid (AA) or U‐46619, a thromboxane A2 analog, caused pressor effects. Moreover centrally injected histamine leads to pressor response by activating central histamine receptors. In the present study, it was aimed to demonstrate interaction between central prostaglandinergic and histaminergic system on central regulation of cardiovascular system by activating prostaglandin cascade with central administration of melittin or AA.

Experiments were performed in male Spraque Dawley rats. Intracerebroventricularly (i.c.v.) injected melittin or AA caused dose‐ and time‐dependent increases in mean arterial pressure (MAP) and decrease in heart rate (HR) as we reported previously. Also, melittin or AA accompanied by 28 % or 56 % increase in extracellular histamine concentration in the posterior hypothalamus, respectively as shown in microdialysis studies. Moreover, H2 receptor antagonist ranitidine almost completely and H3/H4 receptor antagonist thioperamide partly blocked melittin‐evoked cardiovascular effects, whereas H1 receptor blocker chlorpheniramine had no effect.

In conclusion, obtained first data show that there is an interaction between central histaminergic and prostaglandin cascade to central regulate cardiovascular system.

This study was supported by grants from TUBITAK (110O878).
Nucleus tractus solitarii (nTS) reactive oxygen species (ROS) contribute to acute intermittent hypoxia (AIH)‐induced phrenic nerve long‐term facilitation (pLTF)
Authors: Kleiber Allison Christine , Ostrowski Daniela , Kline David D. , Heesch Cheryl M. , Hasser Eileen M.
Published: June 2021

Rats exposed to AIH exhibit cardiorespiratory changes associated with obstructive sleep apnea, such as pLTF, including elevated phrenic (PNA) and sympathetic (SNA) nerve activity that persist ≥ 60 min after the final hypoxic bout (AIH‐pLTF). The nTS in the medulla integrates and modulates primary cardiorespiratory input. Also, ROS modulate nTS neuronal function and are increased by intermittent hypoxia. Therefore, we hypothesized that nTS ROS contribute to AIH‐pLTF. We decreased ROS in the nTS by bilateral microinjection of catalase (CAT, 500 U/ml; 90 nl) prior to each of 10 45‐sec bouts of 10% O2 separated by 5‐min recovery periods (100% O2) in anesthetized, ventilated, vagotomized, and paralyzed rats. AIH alone or with nTS aCSF produced AIH‐pLTF (60 min post‐AIH; integrated PNA amplitude: 451±170% baseline, n=4; int.splanchnic SNA: 183±50% baseline, n=4). Following CAT, increased PNA and SSNA due to each hypoxic bout were preserved. However, CAT attenuated AIH‐pLTF (int.PNA: 0±7% baseline, n=3; int.SSNA: 2±3% baseline, n=2). AIH did not alter arterial pressure or heart rate. Attenuation of AIH‐pLTF by nTS CAT indicates that ROS, likely H2O2, within the nTS normally contribute to AIH‐pLTF. HL98602
Activity of Hypothalamic Paraventricular Nucleus (PVN) Neurons Controls Sympathetic Nerve Activity (SNA) and Respiration in Angiotensin II (AngII)‐Salt Hypertensive Rats: No Role for Local NMDA Receptors (NMDAR)
Authors: Holbein Walter , Herrera‐Rosales Myrna , Toney Glenn
Published: June 2021

Rats consuming high salt (2% NaCl) and treated with AngII (150 ng/kg/min, s.c.) develop hypertension that is dependent on ongoing SNA. In telemetric recordings, we have determined that AngII‐Salt hypertensive (HT) rats also have increased respiration rate. Because PVN neurons innervate sympathetic and respiratory network components, we hypothesized that PVN neuronal activity supports SNA and respiration more in AngII‐Salt HT (n=6) than control (0.4 % NaCl diet, n=9) rats. With ETCO2 set at 5%, mean arterial pressure (MAP) was ~13 mmHg higher in AngII‐Salt HT rats than controls, but burst frequencies of phrenic nerve activity (PNA) were similar. In control rats (n=4), inhibition of PVN (muscimol, 100 pmol in 50 nl/side) had little effect on splanchnic SNA (−7 ± 4 %) or MAP (−1 ± 2 mmHg), but both were reduced (P<0.05) in AngII‐Salt HT rats (splanchnic SNA: −22 ± 3 %, MAP: −39 ± 6 mmHg, n=3). PVN muscimol also reduced PNA bursts (amplitude: −7 ± 4 %, AUC: −15 ± 1 %) and neural minute ventilation (−7 ± 4 %), but only in AngII‐Salt HT rats. Support of PVN neuronal activity by local NMDAR was tested by injection of the antagonist AP5 (200 pmol in 50 nl). In control (n=5) and AngII‐Salt HT (n=4) rats, AP5 had no affect on SNA, PNA or MAP. These data indicate that PVN neuronal activity plays an enhanced role in supporting SNA, inspiratory activity, and MAP in AngII‐Salt HT rats, but this does not depend on local NMDAR. Support: HL102310 & HL088052 (GMT)
4‐phenylbutyrate protects against angiotensin II/DOCA salt‐induced hypertension in a C57BL/6 mouse model of chronic kidney disease.
Authors: Carlisle Rachel , Mohammed‐Ali Zahraa , Dickhout Jeffrey
Published: June 2021

Others have shown that angiotensin II infusion combined with deoxycorticosterone (DOCA)/high salt results in hypertension characterized by both increased total peripheral resistance and plasma volume expansion. We have previously shown that 1.0% NaCl, in this model, results in renal pathology, characterized by proteinuria, intertubular protein casts, renal interstitial fibrosis, and endoplasmic reticulum stress. We hypothesized that treatment with the endoplasmic reticulum stress inhibitor, 4‐phenylbutyrate (4‐PBA), would protect against this model of hypertension‐induced chronic kidney disease. C57BL/6 wild type mice were either untreated, or pre‐treated with 4‐PBA for one week, at a dose of 1 g/kg/day, delivered in the drinking water. Chronic kidney disease was then induced by subcutaneous administration of angiotensin II and DOCA for twenty‐one days, with 0.8% NaCl in the drinking water or 0.8% NaCl with 4‐PBA in the drinking water. Administration of angiotensin II and DOCA high salt resulted in a significant increase in blood pressure, as well as glomerular damage and renal hypertrophy, in the mice. Co‐treatment with 4‐PBA prevented an increase in systolic and diastolic blood pressures, measured indirectly with a tail cuff (CODA System), and reduced the renal hypertrophy associated with kidney damage. Supported by CIHR OSO‐115895.
Posterior communicating artery remodeling in chronic cerebral hypoperfusion
Authors: Matin Nusrat , Neros George , McClain Jonathon , Dorrance Anne McLaren
Published: June 2021

Bilateral carotid artery occlusion (BCAO) is an established model of chronic cerebral hypoperfusion that causes vascular cognitive impairment (VCI). To compensate for the occlusion of the carotid arteries, flow from the vertebrobasilar system may increase, and may alter the structure of the posterior communicating artery (PComA). The aim of this study was to understand how BCAO affects the PComA, and we hypothesized that BCAO would cause outward remodeling of the PComA. 20 week old Long Evans rats had BCAO. Data are mean± SEM, SHAM vs BCAO. 5 weeks later the rats had VCI as evidenced by an increased latency to find the platform (7±0.8 vs 28±1.3 s, p<0.05, ANOVA) in Morris water maze testing and a decrease in novel exploration quotient (0.7±0.03 vs 0.5± 0.02, p<0.05, ANOVA) in novel object testing. Structure of PComAs was assessed by pressure myograph (data shown at 80 mmHg). Lumen diameter was increased in BCAO (148±6.1 vs 441±30μm, p<0.05, ANOVA). Wall/lumen ratio was reduced (0.1±0.01 vs 0.04±0.01, p<0.05, ANOVA) while stress was increased (431±32 vs 1106±160 dynes/cm2, p<0.05, ANOVA) in the BCAO. These data suggests that, increased flow through the PComA causes outward remodeling and begets investigation of the reactivity of the PComA. Modulating blood flow through the PComA may be a therapeutic target for VCI.
Influence of hand dominance on peak forearm blood flow and cardiovascular responses to isometric handgrip exercise in recreationally active racquet players
Authors: Kimmerly Derek Stephen , Hughes Lauren , Quattrocchi Lauren
Published: June 2021

The purpose of this study was to determine if recreationally active racquet players (6.3 ± 2.4 years played) exhibit different responses to isometric handgrip exercise (IHG) between the dominant (D) and non‐dominant (ND) forearms than age‐, sex‐ and upper body aerobic fitness‐matched controls (n=8 per group). Two separate protocols were performed with each forearm: A) 5‐min of arterial occlusion (AO) combined with 1 min of IHG (15% max force), and B) 2‐min of IHG (40% max force) followed by 2‐min of AO. Forearm blood flow (FBF, plethysmography), heart rate (HR) and mean arterial pressure (MAP, Portapres®) data were recorded. Peak FBF was higher in both forearms (p<0.01) of the racquet players than controls (D; 21.2 ± 4.9 vs. 14.9 ± 4.5, ND; 20.2 ± 5.7 vs. 13.3 ± 2.8 ml/100ml/min). During protocol B, HR (D; 23.6 ± 11.7 vs. 15.1 ± 9.0, ND; 22.4 ± 11.9 vs. 12.6 ± 8.0 beats/min) and MAP (D; 128 ± 16 vs. 105 ± 17, ND; 123 ± 23 vs. 96 ± 21 mmHg) increased more during IHG in both limbs of the racquet players (all, p <0.05). MAP was also greater (p <0.05) during post‐exercise AO in both limbs of the racquet players (D; 112 ± 16 vs. 99 ± 13, ND; 111 ± 19 vs. 91 ± 13 mmHg). Compared to similar results from elite racquet sport players, differences in peak FBF, HR and MAP responses to IHG between groups were not isolated to the dominant forearm. These data suggest an absence of a localized training effect to the dominant forearm in recreationally active racquet players.
Circulating and intracellular HSP72 responds acutely to cumulative exercise stress, heat stress, dehydration, and rehydration and may be mediated by collective effects of multiple stress‐specific transcription factors
Authors: Muñoz Colleen X. , Lee Elaine C , McDermott Brendon P , Beasley Kathleen N , Yamamoto Linda M , Emmanuel Holly , Hom Lindsay L , Casa Douglas J , Armstrong Lawrence E , Kraemer William J , Maresh Carl M
Published: June 2021

We have previously shown that protective protein HSP72 responds acutely and chronically during 11d of daily exercise‐heat stress. In this study (n=6, men), we aimed to characterize acute HSP72 and transcription factor responses to 2d of sequential stress and rest periods. In each of 4 repeat visits, participants: 1) passively dehydrated overnight, 2) exercised for 2h with no fluid in a hot and humid environmental chamber, 3) rested and rehydrated for 1h, 4) maximally exercised for 0.5h, and 5) returned after 24h of at‐home recovery and rehydration. We tracked rectal temperature, hydration status, circulating HSP72, and intracellular (flow cytometry) expression of HSP72 and cellular stress‐specific transcription factor NFAT5. We found that circulating and intracellular HSP72 significantly correlated (p<0.05) with physiological manifestations of heat, exercise, and dehydration stresses and recovery/rehydration periods. NFAT5 expression correlated (p<0.05) with measures of hydration status suggesting for the first time that this cellular osmotic stress‐induced transcription factor may reflect whole body hydration status, and contribute to general protective responses during aggregate stress.
Responses of circulating irisin to different exercises in humans
Authors: Mougios Vasileios , Huh Joo Young , Panagiotou Grigorios , Kabasakalis Athanasios , Mantzoros Christos
Published: June 2021

Irisin is a newly characterized myokine that may aid weight reduction and insulin sensitivity. We have recently shown that circulating irisin increases in humans after acute exercise taxing the energy state of muscle. To further explore the types of exercise that upregulate irisin, we measured irisin using enzyme immunoassay in plasma of 30 swimmers before and immediately, 1 h and 24 h after two exercises of different intensities: a set of six 50‐m maximal swims raising blood lactate to 17 mmol/l, and 2000 m of continuous swimming raising lactate to 6 mmol/l. Irisin increased (P < 0.05) only after the 6 × 50‐m trial and then gradually returned to baseline. As a test of reproducibility of the irisin concentration, 14 swimmers performed two identical 6 × 50‐m trials one week apart. Irisin concentration measurements proved highly reliable (intraclass correlation coefficient 0.921). Finally, to test if the blood lactate concentration is a good predictor of the irisin response to exercise, we measured irisin in 14 women who performed whole‐body bilateral vibration training for 6 weeks. Despite raising the blood lactate concentration to only 5 mmol/l, vibration exercise increased irisin significantly both at the first and last training sessions. These data suggest that the response of circulating irisin to exercise depends partly on exercise intensity, although other characteristics of exercise may modulate this response.
Treatment of sciatic nerve injury in one experimental model in rat with Hyperbaric Oxygen Therapy
Authors: Ortiz‐Villegas Miguel , Gama‐Aguilar Jorge Alejandro , Castillo‐Hernandez Maria Carmen , Lara‐Padilla Eleazar , Guevara‐Balcazar Gustavo
Published: June 2021

Participation in sports has increased significantly in our society, the benefits associated with this practice are well documented in young, adults and elderly; interestingly competitive sports have taken on new meaning and intensity in sports has increased as well as the degree in lesions in athletes

One of the more commune injuries in soft tissue is in the lumbar region affecting the intervertebral disks, known as disc herniation. Constantly occur in the athlete that is under great force in your body for an incorrect position. Sports injury showing greater lumbar and sciatica nerve injury are rhythmic gymnastics, football and weightlifting.

Hyperbaric oxygen therapy in sports has been used for fatigue and for some inflammatory lesions that make athlete back to training in a shorter period of time.

The sciatic nerve injured protocol in rat with HBO treatment, is a model that allow to follow the recovery in this lesion with oxygen treatment, we use rat with sciatic nerve lesion with and without oxygen therapy, after 10 session of HBO, the recover of the sensibility of the nerve was faster that the group of rats without this therapy.
Metabolic Profile of Brazilian Elite Boxing Athletes
Authors: Burini Franz Homero , Cruz Igor Freitas , Romano Felipe Freitas , Alves Mateus Freitas , Suárez João Carlos , Ayres Claudio Carlos , Torezan Gabriel Carlos , Teixeira Okesley Carlos , Manda Rodrigo Minoru , Santi Bernardino Carlos , Zampronha Waldir Carlos , Burini Roberto Carlos
Published: June 2021

Elite athletes present a peculiar profile that demands specific performance assessment, including training, dietary recommendations and metabolic stress repercussion.

This study aimed to assess the metabolic impairment regarding nutritional intake, eletrolytes, gluconeogenesis (urea cycle) and muscle damage responses of Brazilian elite boxing athletes.

For RMR evaluation, a portable analyzer was used in 11 athletes (7 males/4 females; 25,5±3yrs); nutritional pattern was analyzed using daily 24h records, with specific software for qualitative datas. They were submitted to blood tests analysis for acute and chronic biochemical markers of stress.

RMR was 1341,2±423,2, higher in males (1496,7±109,2; 1069,2±646,4). Caloric intake was 2038,6±575,2, higher in females (2323,2±786,8; 1874,78±394,6). Qualitative analysis showed lower carbohydrate intake (91% of the athletes). 36% showed high protein intake. Eletrolytes intake, such as sodium and pottasium were abnormal in 100% of the athletes. CK values were abnormal in 90% of the athletes; Urea was elevated in 25% of the athletes. Sodium and potassium were above the DRI, and the serum levels were in normal range (138±18mg/dL; 4± 10.8mg/dL).

The present study evidences abnormalities that should be focused on the multidisciplinary matter, avoiding deleterious atitudes such as dehydration and malnutrition, wich could lead to decrease in performance.
Ketamine (KET) suppresses inflammatory genomic response to acute hypoxic hypoxia (HH) in late gestation fetal ovine kidney cortex
Authors: Chang Eileen I‐Ling , Rabaglino Maria Belen , Richards Elaine M. , Wood Charles E.
Published: June 2021

The purpose of this study was to evaluate the genomic response of fetal kidney cortex under HH with ketamine. Fetal sheep (n=4/group) were chronically catheterized (125 d gestation). Ketamine (3 mg/kg) was administered intravenously to the fetus 10 min prior to the induction of HH (30 min) stimulation or normoxic control (NC). Fetal kidney cortex was collected 24 hrs post‐HH, and mRNA was extracted and analyzed using ovine Agilent 15.5k array. Significant differences in gene expression between treatment (HH and HH+KET) and control (NC and NC+KET) groups were determined with F‐statistics using the limma package (P<0.05) for R. Significantly up‐/down‐regulated genes: HH: 405/813 and HH+KET: 274/208. Enriched biological processes (BP) for the upregulated genes under HH were immune and inflammatory response, cytokine production, and apoptosis. Down‐regulated BP were protein location and transportation, chromosome organization, and mRNA processing. With ketamine, up‐regulated BP were metabolic and cellular processes, cell cycles, and DNA repair. Down‐regulated BP were immune system response, regulation of apoptosis and cell proliferation. In conclusion, 24 hrs post‐HH stimulation increased genomic expression of inflammatory pathways in fetal kidney cortex, but with ketamine, the inflammatory genes were down regulated. Research supported by NIH grants HD033053, HD057561, and DK076541.
The effects of creatine loading on oxygen uptake kinetics during heavy exercise
Authors: Butts Cory L , Adamus Heather , Keller David M , McDonough Paul
Published: June 2021

The kinetics of oxygen uptake during the rest to exercise transition are thought to be modulated by intracellular metabolic processes. Evidence suggests that increased phosphocreatine stores may impact the kinetics of VO2, namely, slowing kinetics due to the now increased ATP buffering capacity. Thus, the purpose of this study was to test the hypothesis that creatine loading would induce slowed VO2 kinetics during heavy leg cycling. Four (N=4) subjects completed three separate exercise bouts of heavy (supra‐lactate threshold) exercise both prior to and following a creatine loading regimen. VO2 was measured continuously at the mouth during exercise and recovery for each bout. During the on‐transition, the total amplitude was reduced following creatine loading (Atot: Control 1.22±0.26 vs Creatine 0.97±0.20 L/min, p = 0.03). The time constant during the off‐transition was also slower during the creatine trials (Tau: Control 36.4±9.3 vs Creatine 53.3±7.6 sec, p = 0.02). These findings suggest that an increase in the temporal buffering capacity of phosphocreatine reduces the on‐transient VO2 as well as, slowing the recovery kinetics of VO2.
Glutamatergic neurotransmission plays a role in BDNF/TrkB.FL‐induced enhancement of functional recovery after cervical spinal hemisection
Authors: Gransee Heather M , Zhan Wen‐Zhi , Bailey Jeffrey P , Sieck Gary C , Mantilla Carlos B
Published: June 2021

Brain‐derived neurotrophic factor (BDNF) acting through full‐length tropomyosin related kinase receptor subtype B (TrkB.FL) plays an important role in neuroplasticity following spinal cord injury. BDNF/TrkB.FL signaling in phrenic motoneurons is both necessary and sufficient to enhance spontaneous recovery of ipsilateral rhythmic phrenic activity after C2 spinal cord hemisection (SH), where premotor drive to phrenic motoneurons is removed. Phrenic motoneuron expression of the excitatory glutamatergic NMDA receptor correlates with the time course of spontaneous functional recovery post‐SH. We hypothesized that increased BDNF/TrkB.FL signaling in phrenic motoneurons enhances recovery of rhythmic phrenic activity after SH via regulation of glutamatergic receptor expression. Rats were injected intrapleurally with AAV7 expressing human TrkB.FL‐FLAG at 3 weeks prior to SH. FLAG was present in the cervical ventral horn region, and AAV7‐GFP expression was specifically in phrenic motoneurons. At 14 days post‐SH, quantitative real‐time RT‐PCR verified human TrkB.FL expression in microdissected phrenic motoneurons. Phrenic motoneuron expression of NMDA increased ~10 fold compared to control, suggesting that increased BDNF/TrkB.FL signaling magnified SH‐induced changes in phrenic motoneuron NMDA expression. Supported by HL96750 and T32‐HL105355, and Paralyzed Veterans of America.
Predicting Susceptibility to Alveolar Flooding in Terms of Critical Cardiovascular Parameters
Authors: Heiner Jacob A. , Sullivan Kyle H. , Henderson Megan E. , Dongaonkar Ranjeet M. , Quick Christopher M.
Published: June 2021

Modest increases in pulmonary interstitial fluid pressure (Pi) can lead to alveolar flooding and respiratory failure. Arising from the complex interaction of microvascular filtration, lymphatic drainage, as well as cardiac, renal and venous function, Pi is typically predicted by mathematical models that are notoriously complex, difficult to interpret, and solvable only by those with advanced programming skills. Therefore, we developed a simple model that predicts Pi as a function of critical cardiovascular parameters. Microvascular filtration into the pulmonary interstitium is characterized by the Starling‐Landis equation, with parameters for protein and water permeability. Lymph flow out of the interstitium is characterized by a lymphatic model with parameters for sensitivity to Pi and central venous pressure. Finally, capillary and central venous pressures are characterized by a standard minimal closed‐loop model with parameters for ventricular compliances and contractilities, arterial and venous compliances, and arterial and venous resistances. By linearization and judicious use of simplifying assumptions, we developed algebraic solutions predicting Pi. This simple model is comprehensive enough to predict the susceptibility of different patient populations to alveolar flooding with such diverse conditions as sepsis, hemodilution, and fluid volume retention with acute heart failure.
Anandamide inhibits proliferation of oral squamous cell carcinoma
Authors: Flynn Andrea N , Schmidt Brian L
Published: June 2021

Cancer patients experience severe, unmanageable pain. Patients often develop tolerance and suffer side effects from opioids. The endocannabinoid system includes the cannabinoid receptors (CBr1, CBr2), the endogenous ligands, anandamide (AEA) and 2‐ arachidonoylglycerol (2‐AG), and their cognate degradative enzymes, fatty acid amide hydrolase and monoacylglycerol lipase (MAGL). Endocannabinoids have recently emerged as potential antitumor agents. However, their effect on head and neck cancer remains undefined. We monitored the proliferation of an oral carcinoma cell line, HSC3, over 24 hours using the xCELLigence Real‐Time Cell Analyzer. We found AEA inhibited proliferation with an IC50 of 1.05 μM. We also found Ki67, a marker of proliferation, was significantly reduced following addition of 5 μM AEA for 24 hours. HSC3 proliferation was also reduced following treatment with PF‐750, an irreversible inhibitor of FAAH. AEA has been shown to activate CBr1, CBr2, and TRPV1. Preliminary experiments with specific inhibitors to CBr1, CBr2, and TRPV1 indicate AEA elicits these anti‐proliferative effects via CBr1 and TRPV1. These results indicate regulation of the endocannabinoid system could be a promising treatment for oral cancer and associated pain.
The evolution of arthropod appendages
Authors: Patel Nipam H.
Published: June 2021

Extensive work in several model species has given us a detailed understanding of the mechanisms that guide the development of animals and plants. One remarkable outcome of these studies is the finding that the genes and pathways that underlie development are well conserved across large evolutionary distances, despite many of the obvious differences in the morphology of these organisms. Comparative approaches are now yielding insights into how conserved genetic pathways are altered by evolution to generate this diversity. Increasingly, the adoption of new species for detailed developmental studies plays a key role in uncovering these evolutionary changes, and these analyses are greatly facilitated by advances in many experimental and analytical techniques. I will discuss the insights obtained in understanding the evolution of body patterning and appendage diversification that have come from the study of arthropod evolution, highlighting our recent work on Homeotic (Hox) genes in the amphipod crustacean, Parhyale hawaiensis.
Teaching and examining medical students on their ability to writing prescriptions using the e‐learning program Pscribe
Authors: Janssen Ben , van Doorn Adriaan
Published: June 2021

Preparing medical students to become expert in prescribing medicines is a major challenge. To provide a better training a network of Dutch academic teachers is developing a new web‐based e‐learning program Pscribe (http://www.pscribe.eu). Pscribe offers students virtual patients to test and practice their pharmacotherapeutic skills. The program is based on a WHO model that uses 6 steps in the process of rational prescribing to solve patient problems. Building cases in Pscribe allows enrichment with a wide variety of text and multimedia elements and includes direct web‐links to the official national registry of medicines and formularies of additional countries. Tracking modules and questionnaires are available that enable teachers to follow individual students in their decision making. The program monitors all drugs that have been studied and selected as well as the time needed for preparation of each step and allows personal correction of the actual lay out of the prescription. Feedback can be given on a group level either directly when making the case or on a personal level after students hand in the assignment for official examination. Data on the performance and appreciation of students for this program will be presented for cohorts of year 1 (n=350) and year 2 (n=350) medical students of the Maastricht University, The Netherlands.
An undergraduate laboratory activity on Molecular Dynamics simulations
Authors: Rodriguez Juan , Goadrich Mark , Messina Troy
Published: June 2021

Recent reports such as Vision and Change [AAAS, 2009] and A New Biology for the 21st Century [NRC, 2009] have highlighted the need for undergraduate biology curricula that emphasize interdisciplinary approaches, and that focus on key concepts and competencies deemed critical for 21st century biologists. Here we describe a laboratory activity that encompasses these recommendations. The activity bridges biology, physics, computer science, and mathematics, addresses the key concept of Structure and Function, and enhances students’ competencies in computational and mathematical modeling. The activity is centered on Molecular Dynamics, a computer simulation method that is now widely used as a tool for determining the structure and function of proteins. The activity takes place over two lab periods of three hours each and can be carried out with two software packages that are available free of charge. The first lab period is aimed at unpacking the principles and methods used in Molecular Dynamics in a way that is accessible to bioscience students. During this period students are guided through simple physics principles and taught rudimentary programming skills in Python (http://www.python.org/) to simulate the motion of a bound atom. In the second period they extend concepts learned in the first to simulate the change in conformation of a modified protein using VMD/NAMD (http://www.ks.uiuc.edu/).
Using 3‐D surface models to evaluate morphological variation in artiodactyl carpal and tarsal bones
Authors: Dunn Rachel Heather
Published: June 2021

The postcranial bones of artiodactyls are widely used in ecomorphological studies, however, there is debate about whether certain elements (such as the astragalus) vary enough in shape to be useful in such analyses. To address this issue, I am interested in two questions: 1) Are some bones more morphologically conserved than others? 2) Can surface area and angular data taken from 3‐D models provide meaningful morphological information in artiodactyls? In this study I looked at three carpal bones (lunar, unciform, and cuneiform) and one tarsal bone (astragalus) in six families of artiodactyls. My results suggest that some bones are more morphologically conserved than others, with the cuneiform and astragalus being less variable morphologically than the lunar and unciform. In addition, I have identified potentially phylogenetically informative features of the lunar and unciform, for example, ruminants have a relatively large facet for the magnum on the unciform than non‐ruminants, and bovids have a relatively larger unciform facet on the lunar than other ruminants. Other characteristics are potentially homoplastic, for example the mountain goat and bighorn sheep share a large scaphoid facet on the lunar with non‐bovids, whereas other bovids have relatively small scaphoid facets. These results suggest that some elements are more useful than others for ecomorphological research.
Artery of Adamkiewicz: Medullary Artery or Radicular Artery?
Authors: Schenk Michael P. , Haines Duane E , Cunningham W. Kyle
Published: June 2021

The artery of Adamkiewicz (A of A), after Albert Adamkiewicz (1882–1921) provides 75–80% blood supply to the spinal cord by way of the anterior spinal artery. Haller in 1754, described this vessel in “Iconum Anatomicarum”, but did not name it. This artery, the “magnus ramus radicularis anterior” or “arteria magna spinalis”, is interpreted as the greater radicular artery. Recently, it has been recognized by other anatomical terms.

From the intercostal a., a spinal a. enters the neural foramen and penetrates the dural root sheath as a radicular a., a medullary a. or both. Consequently, the radicular aa. supply the nerve roots (L. radix, root), and the medullary aa. supplement the anterior spinal a. (75%) while the posterior medullary a. supplement the posterior spinal aa. Contributions to the nerve root by the medullary aa. is minimal.

It has been known for decades the medullary a. supplements the spinal cord and not the radicular a. Anatomical knowledge of (A of A) and its correlation with the intercostal aa. is essential in the prevention of postoperative neurosurgical/cardiovascular procedures, i.e. paraplegia.

Purpose

Clarify arterial structures of the spinal cord, and emphasize anatomical relationships of (A of A).

Method

Extensive literature search, dissection and consultation with both basic science and clinical faculty.
A morphological study of the mandibular foramina in Japanese macaques
Authors: Sato Iwao , Miwa Yoko , Zaizen Tomonori , Sunohara Masataka , Yosue Takashi , Shimada Kazuyuki
Published: June 2021

Introduction

Two foramina and related accessory foramina are present at the labial surface of the mandible in Japanese macaques. However, the correlations between the shape and the location of these foramina haves not been precisely reported. These foramina may indicate the course of the nerve and arterial supply.

Materials and methods

Dental cone beam computed tomography (CBCT) was used to evaluate the morphology of the mandible in Macaca fuscata using the osteological collections of the Dept. of Neurolo, Gross Anat, Kagoshima Univ. Grad. Sch. Med&Dent. This study focused on the morphological features of the foramina and the relationships between the primary foramen and the accessory foramina in the Japanese macaque.

Results

We investigated the multi‐foramina structure of the Japanese macaque mandible using CBCT and observed the morphological structures of the mental foramen and accessory foramina at the labial surface of the mandible using 3D images. The lingual multi‐foramina were also identified at the anterior lingual surface of the mandible. We classified three types of canal from 3D images, a tube‐like shape, a funnel shape, and a branched shape. The two canals were found in six cases (42.9%, 6/14) and contained nerves and blood vessels, which could be observed macroscopically. The nerves and blood vessels in the multi‐foramina supplied the soft tissues of the labial and lingual regions of the mandible.
A Detailed Anatomical Description of a Coiled or a Kinked Internal Carotid Artery in Three Dissected Cadavers
Authors: Kim PilWoo , Romfh John H , Ledwitz‐Rigby Florence
Published: June 2021

Three human cadavers were found to have a rare internal carotid artery (ICA) anomaly during dissection. The first cadaver presented ICA with coiling anterior to transverse process (TP) of atlas (C1). The other two cadavers presented with kinking patterns around the level of transverse processes of second and third cervical vertebrae (C2–3). Coiling and kinking patterns may represent congenital vascular anomalies, arteriosclerotic pathology or fibromuscular dysplasia and may have some relevance to cerebrovascular disease. Kinking and coiling may create disturbance of the flow of the blood to the brain greater disturbance in the flow of the blood to the brain, and/or occurrence of ectasia and aneurismal dilation in the carotid siphon. The ICA is located in close proximity to the TP of the vertebrae, so that neck movements or injuries may affect the ICA. This anatomical relationship may explain the cause of dizziness or fainting individuals.
Clinical Relevance of Diastasis Recti Abdominis Revisited
Authors: Curran Robert
Published: June 2021

Diastasis Rect Abdominis is the separation, not splitting, of the rectus abdominus muscles, it is quantified by the inter‐recti distance.

Diastasis rectus is the most common congenital abnormality of the abdominal wall. It consists of an upper midline protrusion of the abdominal wall between the right and left rectus abdominis muscles. In the adult, the population most often cited with this condition is the postpartum female, although males can also be effected. The separation occurs frequently during pregnancy, either gradually or suddenly, as a result of exertion imposed on weak musculature.

CIGNA's clinical policy states, “Other than its untoward cosmetic appearance, diastasis recti does not lead to any complications that require intervention. Diastasis recti has no clinical significance, does not require treatment and is not considered a true hernia (Jeyarajah and Harford, 2010).

This presentation will re‐investigate the pathophysiology, biomechanics, and literature associated with this condition, possibly drawing different conclusions.
Cutaneous innervation of the ankle: an anatomical study with clinical implications
Authors: Duscher Dominik , Wenny Raphael , Weninger Patrick , Hirtler Lena
Published: June 2021

Introduction

Three nerves innervate the skin in the foot and ankle region. The saphenous nerve supplies the skin on the medial side of the foot, the sural nerve the lateral side and the superficial peroneal nerve the middle part. Due to their course in the vicinity of the medial and lateral malleoli, these nerves are at significant risk during orthopedic interventions.

The aim of this study is to investigate the distal course of the three cutaneous nerves of the ankle and to determine their exact relationship to easy identifiable bony landmarks. The clarification of the orientation of the nerves around the ankle should be beneficial to the reduction of nerve injuries during surgical approaches in this area.

Materials and methods

10 fresh‐frozen and 40 embalmed lower extremities of adults were dissected. The skin and the subcutaneous adipose tissue were removed to expose the underlying neural structures. Appropriate bony landmarks were defined. To locate the exact position of the nerves standardized digital photographs were taken and computer‐assisted measurements were conducted.

Results

The positions of the superficial peroneal nerve, the sural nerve and the saphenous nerve were measured using reference lines: the intermalleolar line and the line from the fifth metatarsal bone to the highest point of the Lisfranc‐line for the superficial peroneal nerve; the line from the lateral malleolus to the upper border of the calcaneus and the line from the fifth metatarsal bone to the highest point of the Lisfranc line for the sural nerve and the line from the medial malleolus to the head of the talus for the saphenous nerve.

Conclusion

The determination of the exact orientation of the nerves around the ankle should help minimizing the nerve injury rate during surgical approaches in this area. With the described measuring method using easy to identify bony landmarks the location of the cutaneous nerves of the ankle region could be defined precisely.

Grant Funding Source: AAA
The role of the tenocyte in tendon ossification in an avian model
Authors: Agabalyan Natasha Anahid , Evans Darrell John Rhys , Stanley Rachael Louise
Published: June 2021

Tendon ossification, the deposit of bone‐like material within tendon tissue, is common amongst our aging population. Often resulting in painful and debilitating conditions, this ossification compounds damage on an already vulnerable tissue. Although tendon is a key component of the musculoskeletal unit, its repair when damaged is poor. Whereas human tendons may undergo this process through injury, age or even disease; the hindlimb tendons of the adult domestic chicken (Gallus gallus) undergo natural ossification. Little is known about the processes underlying this mechanism, yet it would stand to reason that tenocytes play an important role.

The nature of tissue and cell components of the adult ossified avian tendon were analyzed and compared to the developing non‐ossified avian tendon. The role of the tenocyte and its potential plasticity was investigated both in vitro and in vivo. And the existence of multipotent cells within tendon tissue, Tendon Derived Stem‐like Cells (TDSCs), was explored.

Analysis of the adult ossified avian tendon revealed an ossification process comparable to endochondral ossification. Tenocyte plasticity was analyzed and shown to be restricted to a population within tendon issue, TDSCs.

Understanding the role of the tenocyte and the process behind tendon ossification may help prevent ossification occurring in injured or diseased tendon.

Funded by The Anatomical Society

Grant Funding Source: none
Expression of Steroids Sex Hormone Receptors in the External Genitalia of She‐Camels (Camelus dromedarius)
Authors: Inpanbutr Nongnuch , Emam Mahmoud A. , El‐Zoghby Ihab , Bakry Hatem , Bolon Brod
Published: June 2021

To explore the role of hormonal fluctuations on external genitalia function in female camels, the localization and distribution of steroid sex hormone receptors were examined by immunohistochemistry in vestibule, vulva, and clitoris acquired during breeding and non‐breeding seasons. Animals (n=6 per season) were adults (5 – 10 years) with no history of reproductive tract or systemic disease. Estrogen receptor‐a (ERa) was expressed throughout all mucosal and submucosal tissues in all three regions. Progesterone receptor (PR) was expressed throughout the vestibule and along the mucosal surface of the vulva, but it could not be seen in vulvar skin and the clitoris. Androgen receptor (AR) was expressed throughout the vestibule, clitoris and vulvar mucosa. Epithelial expression of ERa and PR was higher during the breeding season, but AR was higher during the non‐breeding season. These data suggest that significant seasonal changes (p<0.05) in sex steroid receptor expression may play a role regulating external genitalia function in female camels.

This paper was supported by grant from the government of Egypt (MAE and EME).