bluuebunny/crossref_metadata_2025 huggingface dataset:
Patterns of morphological integration of the face in boys with autism spectrum disorders
Authors: Aldridge Kristina , George Ian D. , Austin Jordan R. , Duan Ye , Miles Judith H.
Published: June 2021

The neurodevelopmental model of autism spectrum disorders (ASD) suggests that changes in embryonic development result in the spectrum of ASD phenotypes, including distinct facial morphologies. We hypothesize that the facial phenotype in ASD results from differences in morphological integration (MI), the quantitative assessment of developmental relationships among anatomical regions of the face. The purpose of this study is to determine the effects of ASD upon covariance patterns of facial measures to increase our understanding of how this disorder alters developmental mechanisms underlying the emergence of ASD phenotypes. We statistically compared patterns of MI of boys with ASD (N=70) with those of typically‐developing boys (N=31), aged 8–12 years using 3D landmark coordinate data collected from 3D digital stereophotogrammetric images. Our results demonstrate statistically increased MI in ASD, with the greatest differences observed in the upper face. These findings suggest differential effects of ASD on the various regions of the developing face, indicating alterations of formation and growth of the face during early development.

Work supported in part by ASDRP W91ZSQ8006N604 and the Simons Foundation.
The role of microRNA‐208a in cardiac hypertrophy induced by aerobic physical training
Authors: Soci Ursula Paula Reno , Fernandes Tiago , Rosa Kaleizu T. , Irigoyen Maria Claudia , Phillips M. Ian , de Oliveira Edilamar Menezes
Published: June 2021

The miRNA‐208 regulates gene expression of slow myosin heavy chain(MHC)in heart. We investigated the role of miRNA‐208 and its target Thyroid Hormone Receptor Associated Protein 1 (THRAP1) in aerobic training (AT) induced cardiac hypertrophy (CH). Female Wistar Rats(n=21)into 3 groups: sedentary (S), Trained 1(T1): swimming sessions of 60′, 5 days/week (10 weeks). Trained 2(T2): similar to T1 until 8th week, on the 9th week the rats swan twice/day and on the 10th week 3 times/day. Were assessed: heart rate (HR), blood pressure (BP) by direct measurement, VO2max by ergoespirometry, soleus citrate synthase (CS) activity by spectrophotometer, ventricular function and CH by echocardiography (EGC), CH by LV/BW ratio (mg/g) and cardiomyocyte diameter (CD), gene expression of atrial natriuretic factor (ANF), skeletal α‐actin, α‐MHC and β‐MHC and miR‐208 by Real‐time PCR and protein content of α and β‐MHC and THRAP1 by Western Blott. AT decreased HR and increased VO2max (11%/T1 and 15%/T2) and CS (47%/T1 and 105%/T2). LV/BW ratio increased 13%/T1 and 28%/T2 and CD increased 20%/T1 and 30%/T2. Gene expression of skeletal α‐actin decreased 53%/T2. The E/A ratio increased and isovolumetric relaxation time and myocardial performance index decreased. MiR‐208 expression decreased 62%/T2 with a decrease of β‐MHC and an increase of α‐MHC gene expression(β/‐71%,α/+37%) and protein content(β/−45 % α/+72%) in T2. THRAP1 protein increased 40%/T2 compared with S. APT induced physiological CH and improved diastolic function. MiR‐208a and β‐MHC expression decreased and THRAP‐1 increased. The decrease of miR‐208 was opposite to disease and can be a therapeutic strategy in pathological CH.

Support: FAPESP/CAPES/NIH
Feel the Burn, then Feel the Death. ExoU as a Phospholipase
Authors: Bord E. , Farrell Z. , Heur W. , Keslin W. , Laughlin R. , Leedom H. , LeMense A. , Lucre S. , Marable C. , Mielke S. , Olk S. , Orozco C. , Rosio C. , Schubert J. , Smith S. , Wade G. , Weeden M. , Sampe D. , Frank D. W.
Published: June 2021

A major cause of infection‐related deaths in immunocompromised patients is the protein toxin ExoU, encoded by the bacterium Pseudomonas aeruginosa. The Brown Deer SMART (Students Modeling A Research Topic) Team has modeled ExoU using 3D printing technology to have a better grasp on how the toxin interacts with eukaryotic cells. P. aeruginosa uses a type 3 secretion system (T3SS) to inject toxins including ExoU into the cell to disrupt its functionality. The T3SS is a needle‐like structure comprised of proteins that allow the bacterium to transfer effector proteins into innate immune cells. ExoU travels through the T3SS using a chaperone protein (SpcU). Once inside the eukaryotic cell, ExoU interacts with ubiquitin, where it refolds into an active potent phospholipase that breaks down cellular membranes using Ser142 and Asp344 as the catalytic amino acids. The exact mechanism is unknown but the C‐terminus (residues 580–683) helps in targeting the membrane, allowing ExoU to break it down. P. aeruginosa is able to freely reproduce inside the environment of the host organism as the immune system is not able to compensate for the infected cells and bacterium. Left unchecked, this infection will prove fatal. Research is being conducted to create an ExoU inhibitor to reduce the deaths it causes in patients with compromised immune systems. Supported by a grant from NIH‐CTSA UL1RR031973.
Binge drinking, blood pressure, and duration of physical activity as risk factors for cardiovascular disease in college students
Authors: Dubois Danielle Marie , Morrell Jesse Stabile
Published: June 2021

Inadequate physical activity (PA) and pre‐hypertension (PH) are known risk factors for cardiovascular disease (CVD), however, their relationship with college binge drinking has not been closely examined. Data from the ongoing cross‐sectional College Health and Nutrition Assessment Survey were collected between 2005–2012. Introductory nutrition students (n=5604) reported PA and drinking behavior via questionnaire; blood pressure (BP) was measured via automated cuff following standard protocol. The duration of daily PA was categorized as none, <30, 30–59, 60–89, or ≥90 min; BP was categorized as PH (≥120/85) or normotensive (<120/85). Fifty‐two percent of students reported binge drinking within the last two weeks. Male binge drinkers tended to be more likely than non‐binge drinkers to spend ≥90 min/day engaging in PA (37% vs 32%, p=.07). Female binge drinkers were more likely to spend 60–89 min/day engaging in PA vs non‐binge drinkers (36% vs 31%, p<.01). Male binge drinkers were more likely to have PH than non‐binge drinkers (59% vs 45%, p<.01); no difference in prevalence of PH among females were observed. Data suggests that binge drinking is associated with longer duration of PA in all students, and PH is more common among male students that report binge drinking as compared to those that report no binge drinking. Supported by NHAES grant NH00541.
Yuja extract ameliorates DSS‐induced colitis and induces colon cancer cell death by reducing COX‐2 expressions
Authors: Hwang Jin Taek , Sung Mi Jeong
Published: June 2021

In this study, we first evaluated the inhibitory effect of 70% ethanol extract of Yuja (Citrus Junos Tanaka) on colitis and colorectal cancer cells. Yuja extract abolished the LPS‐induced secretions of nitric oxide, tumor necrosis factor‐alpha, and interleukin‐6 in RAW264.7 macrophages. We further investigated these anti‐inflammatory effects using experimental colitis models. Colitis was induced by 3% dextran sodium sulfate (DSS) in BALB/c mice. Oral administration of Yuja extract reduced the colon shortening induced by DSS. Cyclooxygenase‐2 (COX‐2) expression was also decreased in colon tissue after Yuja extract administration. Next we also evaluated whether Yuja extract exerts an anti‐tumor activity in colorectal cancer cells, because ulcerative colitis has been believed to be a risk factor of colorectal cancer. Yuja extract also accelerated cell death and apoptotic body appearance in HT‐29 colorectal cancer cells. This antitumor activity was accompanied by reducing iNOS and COX‐2 expressions. Taken together, Yuja extract may be useful for preventing colitis and colorectal cancer by reducing COX‐2 expressions.

This work was supported by the Yuja Food Cluster Co., Ltd.
Olanzapine impairs glucose and lipid metabolism in differentiated murine adipocytes
Authors: Kang Hye Won , Hong Ki‐Bae , Im Suji , Park Yooheon
Published: June 2021

Atypical antipsychotics such as olanzapine have become the primary choice of medication for treating psychosis. An unfortunate side effect of these medications is weight gain, a condition often associated with the development of metabolic syndromes. To elucidate the mechanism of impairing lipid and glucose metabolism, we examined the glucose and lipid metabolism in olanzapine treated 3T3‐L1 cells. 3T3‐L1 pre‐adipocytes were differentiated by the addition of insulin (1μg/ml), dexamethasone (1μM), IBMX (10mM), and 10% FBS in DMEM medium. Cells were treated with various concentrations of olanzapine (0, 5, 10 and 20μM). Lipid accumulation was estimated by measuring intracellular triacylglyceride contents. Level of mRNA and protein expression of the lipid metabolism related transcription factors were measured by RT‐PCR and Western Blot analysis, respectively. Cells treated with olanzapine showed significant and dose dependent increase in lipid accumulation. mRNA level of fatty acid synthase (FAS), acetyl‐CoA carboxylase (ACC), uncoupling protein‐2 (UCP‐2), and peroxisome proliferator‐activated receptor‐γ (PPAR‐γ) in the cells treated with olanzapine were significantly increased in dose dependent manner.
Lysophosphatidic acid regulates SPHK1 and GPR68/OGR1 in human gingival fibroblasts
Authors: Cerutis D. Roselyn , Weston Michael D. , Nunn Martha E. , Ogunleye Afolabi O. , McVaney Timothy P. , Headen Karmel V. , Miyamoto Takanari
Published: June 2021

Our laboratory has established that lysophosphatidic acid (LPA) regulates the biology of human oral fibroblasts, and that gingival fibroblasts (GF) express the LPA1–5 receptor subtypes. We have also shown that LPA controls immediate‐early gene transcription of inflammatory cytokines, and their receptors and regulators in GF; therefore, we hypothesized that LPA would modulate transcription of the isozymes SPHK1 and/or SPHK2, which make sphingosine‐1‐phosphate (S1P). S1P is under much investigation as a major regulator in immunity and inflammation. Critically, S1P is a primary controller of human neutrophil responses (such as chemotaxis, degranulation, and oxidative burst), making it highly relevant to the inflammation seen in periodontal disease. After GF treatment with 1 × 10−5 M 18:1 LPA for 2h or 8h, RNA was extracted. Agilent Whole Human Genome Oligonucleotide Microarray analysis revealed that LPA consistently and significantly upregulated SPHK1 versus control (3.9 ± 0.5 and 4.7± 0.7 fold at 2h and 8h, respectively). Of great interest, the proton‐sensing receptor GPR68/OGR1 was highly induced: 7.3 ± 4.0 fold (2h) and 18.8 ± 12.8 fold (8h). LPA1–3 can all dimerize with GPR68 /OGR1, which is linked to COX‐2 and inflammatory cytokine induction in other systems. QRT‐PCR was performed to validate the microarray results, using 3 biologic replicates for four periodontal disease‐relevant up‐regulated genes at the 8h time point. These results (level of significance, p < 0.01) provide preliminary evidence that LPA likely exerts complex actions during periodontal inflammation by inducing S1P production and through LPA1–3 hetero‐dimerizing with GPR68. Support: Health Future Foundation (D.R.C).
Topical application of Serotonin + L‐NAME in vivo induces inward remodeling of the rat cremasteric 1A arteriole via a mechanism that is antagonized by the addition of cystamine, a competitive inhibitor of transglutaminase II
Authors: Foote Christopher , Ramirez‐Perez Francisco I. , Hill Michael A. , Meininger Gerald A. , Martinez‐Lemus Luis A.
Published: June 2021

A 4 hr. exposure of isolated vessels, ex vivo, to a variety of vasoconstrictors results in decreased passive diameter: a process termed inward remodeling. In this study we sought to replicate these observations in intact 1A cremasteric vessels in anesthetized rats using intravital microscopy. Surprisingly, neither Angiotensin II (Ang II) + Norepinephrine (NE) alone or in combination with L‐NAME and/or indomethacin nor Endothelin‐1 (ET‐1) + L‐NAME were able to maintain 4 hrs. of constriction and induce remodeling of the cremasteric 1A arteriole. In contrast, Serotonin (5‐HT) + L‐NAME consistently maintained a 4 hr. constriction of the 1A arteriole and led to inward remodeling (a reduction in the maximal passive diameter induced by exposure to papaverine + adenosine + sodium nitroprusside). Furthermore, it was observed that a 10 min. application of 5‐HT + L‐NAME was sufficient to result in an 11.28 % +/− 1.9 reduction in the passive diameter. When the transglutaminase II (TG2) inhibitor, cystamine, was added to the superfusate, arteriolar constriction to 5‐HT + L‐NAME was attenuated and inward remodeling was abolished. These results suggest that protein cross‐linking, via TG2, may play a role in the process of 5‐HT induced inward remodeling in vivo. (NIH HL‐088105 to LAM and 1P01HL095486 to GAM).
Heart rate, PR‐interval and QT‐interval responses to 24hrs hyperoxia in Bmp2 and Bmp4 heterozygous mice
Authors: Howden Reuben , Cooley Ian , Van Dodewaard Caitlin , Arthur Susan , Cividanes Samantha , McCann Kelly , Gladwell Wes , Martin Jessica , Scott Gregory , Ray Manas , Mishina Yuji
Published: June 2021

Background

Prolonged hyperoxia (96hrs) leads to adverse murine cardiac responses, but clinical therapy with high O2 levels is often short‐term. Therefore, understanding susceptibility to short‐term hyperoxia exposure is clinically important. Bone morphogenetic proteins (BMP) 2 and 4 are important in cardiac development and may influence cardiac responses to hyperoxia. We investigated cardiac responses to 24hrs hyperoxia in Bmp2+/− and Bmp4+/− mice.

Methods

Bmp2+/−, Bmp4+/− and WT littermates were exposed to 24hrs hyperoxia (100% O2). ECG was recorded before and during exposure.

Results

No differences in baseline HR, QTcB, QRS, ST‐I, PR‐I, P‐wave duration (P‐d) or PR‐I/P‐d were found (P >;0.05). After ~18hrs hyperoxia, significant decreases in HR and increases in PR‐I and P‐d were found in Bmp4+/− mice only. Also, there were reductions in QTcB in the WT, but not in Bmp2+/− or Bmp4+/− mice after 17hrs hyperoxia (P <0.05).

Discussion

Results suggest that developmental effects of Bmp2 or Bmp4 disruption did not affect basal cardiac function or were not detectable by ECG. During hyperoxia, Bmp4+/− mice were susceptible in terms of HR and atrioventricular conduction responses, which may have important implications for patients treated with O2 who also harbor Bmp4 mutations. Conversely, QTcB in Bmp2+/− or Bmp4+/− mice was not affected by hyperoxia, unlike WT mice, suggesting a protective effect in these genotypes.
Isolated Vascular Smooth Muscle Stiffness as a Common Mechanism to the Increased Aortic Stiffness of Aging and Hypertension
Authors: Sehgel Nancy Lisa , Zhu Yi , Sun Zhe , Hong Zhongkui , Hunter William C. , Vatner Dorothy E. , Meininger Gerald A. , Vatner Stephen F.
Published: June 2021

Increased vascular stiffness is common to both hypertension and aging, and is thought to be due to changes in the extracellular matrix. We have previously found that aortic stiffness was increased in the aorta of aging monkeys, and that isolated vascular smooth muscle cell (VSMC) stiffness, as assessed by atomic force microscopy (AFM), was increased from 12.8±0.3 kPa to 41.7±0.5 kPa. To determine whether this was unique to aging, we examined aortic stiffness in spontaneously hypertensive rats (SHR). In SHR, mean aortic pressure (123±4 mmHg) was increased compared to control Wistar‐Kyotos (96±6 mmHg), as did in vivo aortic stiffness (7.65 mm/ms versus 3.27 mm/ms). In order to determine if a component occurred at the level of the VSMC, we used AFM nanoindentation to measure VSMC stiffness, and also found it increased in SHR compared to WKY (29.2 kPa versus 13.8 kPa). This suggests that increased stiffness is not simply due to changes in the extracellular matrix, but an important component occurs at the level of VSMCs. Since this mechanism is observed in both aging and hypertension, it appears that it is a general mechanism mediating increased vascular stiffness.
Simvastatin reduces hypertensive and vasoconstrictor effects of angiotensin II in normotensive rats.
Authors: Ufnal Marcin , Aleksandrowicz Marta , Drapala Adrian , Kozniewska Ewa
Published: June 2021

Accumulating evidence suggests that beneficial effects of statins may depend on their interaction with angiotensin II (Ang II). We checked whether orally given simvastatin affects hemodynamic and vascular responses to Ang II in rats. We maintained Sprague Dawley rats on either tap water (control) or water containing simvastatin for four weeks. Subsequently, we measured arterial blood pressure and heart rate at baseline and after iv infusions of Ang II. Vascular response to Ang II was studied in vitro in the organ chamber on isolated, pressurized femoral artery branches. We found no significant difference between the groups in mean arterial blood pressure and heart rate at baseline. Intravenous infusion of Ang II at the smallest dose causing pressor response in controls failed to affect blood pressure in the simvastatin group. Arteries of simvastatin treated rats showed decreased vasoconstriction to Ang II in comparison to controls. In conclusion, the study shows that simvastatin reduces the hypertensive response to Ang II. This may be mediated by diminished responsiveness of arteries to Ang II. The study was supported by the Polish Ministry of Science and Higher Education, Iuventus plus grant.
β‐amyloid peptide disturbs hypothalamic glucoregulation and induces hepatic insulin resistance
Authors: Isabel Arrieta‐Cruz , Gutiérrez‐Juárez Roger
Published: June 2021

β‐amyloid peptide (Aβ) is the main component of amyloid plaques, an established hallmark of Alzheimer's disease (AD), responsible in part for the progressive deterioration of cognitive and behavioral functions in AD. Available evidence suggests that hyperglycemia and insulin resistance, two hallmarks of type 2 diabetes mellitus (DM2), are factors that contribute to accelerating the appearance of AD through the impairment in the modulation of Aβ production and degradation. The mediobasal hypothalamus (MBH) has emerged as a center directly involved in the control of insulin action, glucose homeostasis and energy balance. Although a number of in vivo neurotoxic actions of Aβ have been described, a central, direct effect of Aβ on insulin action has not yet been reported. Here we examined the hypothesis that Aβ exerts an acute neurotoxic effect in the MBH resulting in the impairment of insulin action. Thus, we performed central (MBH) infusions of Aβ in normal young Sprague‐Dawley male rats during basal pancreatic insulin clamps to assess insulin action and glucose kinetics. Our results indicate that the acute infusion of Aβ, but not a scrambled‐control (SCR) peptide, decreased hepatic insulin sensitivity indicated by a failure of insulin to efficiently suppress hepatic glucose production (GP), without modifying glucose disposal. The % inhibition of GP was 9.8 ± 2.7% Aβ group vs. 28 ± 4.6% SCR group, while GP was 11.2 ± 0.6 vs 9 ± 0.3 mg/kg/min, respectively. These studies reveal a deleterious action of Aβ in the MBH that impairs insulin action, a novel neurotoxic effect of Aβ.

Funding: NIDDK DK45024 to R. G.‐J.
Identification of phenotypic adaptations to exercise training across mouse strains classified as high and low responders
Authors: Avila Joshua James , Kim Seung Kyum , Massett Michael P.
Published: June 2021

Exercise training responses vary across inbred strains of mice. The purpose of this study was to identify phenotypic adaptions to exercise training across mouse strains classified as high and low responders. Male mice from two inbred strains, a high responding strain, SJL/J, and a low responding strain, C57BL/6J, were assigned to one of the following groups (n ≈ 6/group): sedentary, low ‐ or high‐intensity exercise training for 4 or 8 wks. High intensity training consisted of treadmill running at 85% max. Low intensity training was performed at 65% max. Changes in time and work during a graded exercise test (GXT) were used to determine training responses. C57BL/6J exhibited a significant main effect for training intensity, but not training duration for changes in time and work. SJL/J showed no significant main effect for duration or intensity for change in work. For the change in time in the SJL/J mice, there was a significant effect of training intensity and a significant intensity*duration interaction. Protein analysis for PGC‐1alpha on hindlimb muscle from C57BL/6J indicate that PGC‐1alpha levels were highest in mice that performed four weeks high‐intensity exercise followed by four weeks low‐intensity with the lowest levels being in eight week hi‐intensity mice. These data suggests that intensity has the biggest influence over training capacity.

Supported by NIH grant HL085918 to MPM, William Townsend Porter Pre‐doctoral Fellowship from the American Physiological Society to JJA, Student Research Grant from the Sydney and JL Huffines Institute for Sports Medicine and Human Performance to JJA.
Myocardial IL‐6R expression and IL‐6 signaling following exercise
Authors: McGinnis Graham Ripley , Barberio Matthew , Ballmann Christopher , Peters Bridget , Quindry John , Amin Rajesh
Published: June 2021

Interleukin‐6 (IL‐6) is produced in and released from skeletal muscle during contraction. IL‐6 activity is regulated by a signaling complex with the IL‐6 receptor (IL‐6R). Basal expression of IL‐6R is minimal, but increases with exercise in skeletal muscle. The purpose of this study was to determine if exercise increases myocardial IL‐6R expression, and increases phosphorylation of signal transducer and activator of transcription (STAT3) in the heart. Wild type (WT, n=18) and IL‐6 knock‐out (KO, n=12) mice were grouped as sedentary or exercised. Mice were habituated to treadmill exercise for 4 days increasing from 10–40 min bouts. After 1 d rest, mice performed 3 d exercise for 60 min at 18 m/min. Sedentary animals had equal treadmill time at 0 m/min. Mice were sacrificed pre‐, 0, 30 and 60 min post exercise. Heart, gastroc and plasma were collected, snap frozen and stored at −80°C. Samples were normalized for protein concentration and probed for IL‐6R and P‐STAT3. IL‐6R increased post‐exercise in gastroc (~140%), heart (~250%) and plasma (~300%) in WT only. P‐STAT3 in gastroc increased (~18%) in both WT and KO. Minor increase in P‐STAT3 in heart (~10%) was only in WT. Further study is needed to confirm present findings. It appears exercise causes IL‐6/IL‐6R mediated STAT3 activation in the heart. Findings suggest that alternative cytokines may compensate for lack of IL‐6 in the KO mice. [JCQ Level 3 IGP 2011]
A comparison of fiber specific and whole muscle homogenate markers of muscle activation following HIT and END exercise
Authors: Scribbans Trisha D , Mitchell Andrew S , Vorobej Kira , Simpson Craig A , Quadrilatero Joe , Gurd Brendon J
Published: June 2021

Muscle activation following a bout of high‐intensity interval training (HIT) and endurance exercise (END) was examined in recreationally active men (n=6; age, 19.3±1.4 yrs; VO2peak, 51.9±5.1 ml/min/kg; WRpeak, 264.1±41.6 W). Muscle biopsies were obtained at rest and immediately following a bout of HIT (8 20s intervals, separated by 10s rest, at 170% WRpeak; 481.4±60.1 W) and END (30min of steady state cycling at 65% of WRpeak; 173.7±29.8 W). Bouts were performed a minimum of 1 week apart in random order. MHC expression was identified via immunofluorescent staining while glycogen content was determined via PAS staining. Changes in phosphorylated AMPK (p‐AMPK) and mTOR (p‐mTOR) were determined in whole muscle homogenates via western blotting. Changes in glycogen content were only examined in type I and IIA fibers as only 8 and 16 of 24 samples contained IIX and IIAX fibers, respectively. Significant (p<0.05) glycogen depletion was observed in both type I (HIT −55% END −42%) and IIA (HIT −49%; END −72%) fibers following both protocols but was greater (p<0.05) in type IIA fibers following END than HIT. Interestingly, whole muscle markers of activation, p‐AMPK and p‐mTOR were only elevated (p<0.01) following END (HIT: p‐AMPK, p=0.34; p‐mTOR, p=0.50). These data suggest that use of p‐AMPK and p‐mTOR as markers of whole muscle activation may not be appropriate following HIT. This research was supported by NSERC.
RENAL ISCHEMIA REPERFUSION INDUCED CARDIAC HYPERTROPHY: CONTRIBUTION OF EXTRACELLULAR MATRIX ON CARDIAC REMODELING
Authors: Silva Rogerio Cirino , Sonoda Mayra Trentin , Ramos Marcela Sorelli Carneiro
Published: June 2021

Objectives

Cardiac fibroblasts play a crucial role in cardiac function that is directly linked to extracellular matrix production. The focus of this study was to investigate the modulation of the extracellular matrix during cardiac hypertrophy induced by renal ischemic reperfusion.

Methods

C57bl/6J mice were subjected to unilateral 60 min kidney ischemia, followed by reperfusion of 5, 8, 12, 15 and 20 days. Levels of urea were measured to confirm renal failure, real time PCR, western blotting and FTIR (Fourier Transform Infra Red) were used to molecular analysis.

Results

Urea levels were increased in groups 5, 15 and 20 days (P<0.05). The group 15 days showed a significant increase in heart weight/body weight ratio when compared to sham group. MMP2 levels showed increased at 12 days of reperfusion (p<0.05), but Collagen levels were not altered in this cardiac hypertrophy model.

Conclusion

First, the results indicate that renal ischemia/reperfusion model is able to induce a transient cardiac hypertrophy in mice. This hypertrophy was not accompanied to modulation on collagen levels. Financial Support: FAPESP, CNPq and UFABC.
Integrating Anatomical Education into a Residency Curriculum of a Surgical Specialty
Authors: Cundiff Geoffrey
Published: June 2021

Anatomy is a cornerstone of modern medical education yet there is a continuing trend toward decreasing the time devoted to cadaveric dissection during medical school. Has the decrease in its use negatively impacted student's anatomical foundation? This is an especially relevant question for postgraduate education in surgical specialties, where an understanding of the pertinent anatomy is vital.

Medical students entering residency training in obstetrics and gynecology often do not have an adequate understanding of pelvic anatomy. In fact, the recent growth in professional development courses employing cadaveric dissections is a testament to practicing gynaecologists recognition of inadequate teaching of pelvic anatomy. Designing a program to address this knowledge deficit is a worthwhile aim, especially if it includes elements that will maximize the utility of anatomical knowledge in surgical practice. Learners need a three‐dimensional understanding of pelvic anatomy that is transferable to pelvic surgery. Such a program should have clinical correlations, and develop production sets that are foundational to gynaecologic procedures. Lastly, repetition is essential to knowledge retention.

A multimodality approach to teaching surgical anatomy is the most practical means to achieve these pedagogical goals. Clay modeling is a three‐dimensional technique that serves as an adjunct to lectures to refresh anatomy knowledge in residents entering an Obstetrics and Gynaecology program. The objective of the clay‐modeling session is to allow residents to conceptualize and build the anatomic structures of the pelvis in 3 dimensions using pre‐cut clay structures and a bony pelvis. While the lectures and supplementary teaching methods, are useful to build on the foundation provided by the clay‐modeling lab, the value of cadaveric dissection cannot be overstated. For Gynaecology, both vaginal and laparoscopic cadaver labs should be included, and can be achieved in sequence on the same cadaver. Repetition is the key to sustaining long‐term learning. Perhaps the best way to achieve this is by creating a clinical learning environment that highlights the importance of anatomical considerations and creates opportunities for cognitive reinforcement. Ultimately, the most important approach is to define it as a core competency required for advancement, and to develop discriminating means to test for it. As postgraduate accreditation bodies increasingly adopt competency‐based curricula, the need for valid methods of assessing anatomical competency becomes more pressing.
Kupffer Cell p38 MAPK Drives Hepatic Damage and Pulmonary Inflammation after Intoxication and Burn Injury
Authors: Chen Michael , O'halloran Eileen , Kovacs Elizabeth
Published: June 2021

Alcohol intoxication precedes half of adult burn injuries and leads to worsened clinical outcomes. We recently reported intoxication before burn alters the gut‐liver axis which drives pulmonary inflammation through interleukin‐6 (IL‐6) and we now seek to determine the mechanisms of this aberrant response. C57/BL6 mice were given ethanol (1.2 g/kg) by oral gavage 30 min prior to a 15% scald burn. Depletion of Kupffer cells (KCs) was achieved with clodronate liposomes (0.5 mg/kg). KCs were isolated and analyzed for p38 MAPK ± LPS stimulation (100 ng/ml) and p38 inhibition (SB203580, 20 μM). In vivo p38 inhibition was achieved via intraperitoneal injection of SB203580 (100 mg/kg) 30 min after injury. The absence of KCs attenuated hepatic damage as seen by decreases of 53% (p<0.05) in serum ALT, 37% (p<0.05) in hepatic triglycerides, as well as a 77% reduction (p<0.05) in serum IL‐6 levels compared to intoxicated burned mice receiving empty liposomes. This mitigation of hepatic damage was associated with a 34% decrease (p<0.05) in pulmonary neutrophil infiltration and attenuated alveolar wall thickening. KC's isolated from intoxicated burned mice had a 2‐fold (p<0.05) and 3‐fold (p<0.05) elevation of baseline and LPS‐stimulated p38 activation, respectively. This corresponded to a 2‐fold (p<0.05) increase in IL‐6 which was decreased by 80% (p<0.05) when p38 was inhibited. In vivo p38 inhibition after injury conferred nearly identical hepatic and pulmonary protection after the combined injury as mice depleted of KCs suggesting p38 signaling in KCs may serve as a therapeutic target in this common clinical scenario. This work was supported by NIH R01AA012034 (EJK), F30AA022856 (MMC), T32AA013527 (EJK) and the Falk Foundation.
Inhibition of hydrogen peroxide prevents the development of VEGF‐induced angiogenesis in portal hypertensive rats
Authors: Duan Ming , Qin Jun , He Yue , Luo Meng , Huang An
Published: June 2021

Aims

To determine specific effects of inhibiting hydrogen peroxide (H2O2) on splanchnic hemodynamics and angiogenesis in portal hypertension of rats.

Methods: Portal hypertensive models were induced by partial portal vein ligation (PPVL). The rats then, received either polyethylene glycol‐catalase (H2O2 inhibitor; PPVL‐PEG) or vehicle (PPVL‐Veh) for 8 days. The hemodynamic studies were performed using colored microsphere technique. Intestinal expressions of VEGF/VEGF‐2 and CD31 (angiogenesis markers) was determined by western blotting, and levels of H2O2 were measured by the Hydrogen Peroxide Assay Kit.

Results: Portal pressure was significantly higher, associated with lower splanchnic arteriolar resistance in PPVL group (14.4±0.9 and 11.2±1.1mmHg) than those of normal rats (6.1±0.4 and 35.1±4.0 mmHg). Upregulation of VEGF/VEGF‐2 and CD31 in the intestine/mesentery of PPVL group was indicative of angiogenesis, which consequentially, initiated great collateral vessel formation, leading to the development of splanchnic hyperdynamic circulation. Moreover, levels of H2O2 in the intestine and mesentery of PPVL group of rats were significantly higher than those of normal rats. These alterations were significantly alleviated after treatment of portal hypertensive rats with PEG‐catalase, suggesting a key role of H2O2 in the responses.

Conclusions

H2O2 plays an important role in the pathogenesis of portal hypertension, characterized as the potentiation of splanchnic neovascularisation and development of splanchnic hyperdynamic circulation.
Walnut Consumption Changes the Relative Abundance of Bacteroidetes and Firmicutes in the Gut
Authors: Byerley Lauri , Ponder Monica , Lorenzo Brittany , Banks Shelia , Taylor Christopher , Luo Meng , Blanchard Eugene , Welsh David
Published: June 2021

Eating walnuts is associated with health benefits that include reducing the risk of heart disease and cancer. These chronic diseases are linked to changes in the gut microbiome; for example, a shift in the abundance of Firmicutes (F) to Bacteroidetes (B). Food is also known to modulate gut microbes. One way walnuts may exert their health benefits is by altering the gut microbiome. This study investigated walnut's effects on the two major gut bacterial phyla, F and B. Male Fischer 344 rats (n=20) were randomly assigned to one of two diets for 6 or 13 weeks: 1) walnut‐enriched (W) containing approximately 2 servings, and 2) replacement (R) in which walnuts were substituted by corn oil, additional protein and fiber. After 6 or 13 weeks, fecal samples were collected from the descending colon, the DNA isolated and analyzed. The V3 hypervariable region of 16S rRNA gene was deep sequenced on an Illumina MiSeq. The data were analyzed using the UPARSE and QIIME pipeline. Body weight (W: 340+24 g vs R: 340+24 g) and food intake (W: 15.4+2.6 g/day vs R: 14.9+2.0 g/day) did not differ significantly. F and B were the most abundant phyla present regardless of diet and constituted about 90% of the microbes detected. The length of time the diet was consumed did not change the relative abundance of F and B. However, inclusion of walnuts in the diet significantly altered the relative abundance of F and B, increasing the ratio of F to B (W: 3.2+0.15 vs R: 1.8+0.18, p<0.002). These data suggest a new mechanism, changing the gut microbial environment, by which walnuts may exert their beneficial health effects. (Support: California Walnut Commission)
Mechanisms of Ferritin Iron Mobilization in Macrophages and Hepatocytes
Authors: Tu David , Greenberg Whitney , Tran Ketson , Soriano Cesar , Linder Maria
Published: June 2021

We and others have demonstrated that to retrieve iron stored in ferritin, this large hollow protein (which carries iron‐ferrihydrite crystals) moves into lysosomes, where proteases must degrade its protein “shell” so that the iron may be solubilized and reenter the cytosolic labile iron pool. We have investigated two steps in this process, using hepatocyte and macrophage cell culture models as well as rodents, the first being that leading to ferritin autophagy by lysosomes, the other iron transfer from lysosome to cytosol. It has long been known that a portion of cellular ferritin contains covalently linked oligomers. To test whether these might be involved in ferritin autophagy to lysosomes, we isolated ferritin from liver and spleen of rats pre‐iron‐loaded to induce ferritin, after 5‐days of subsequent injections with saline (control), or erythropoietin (EPO) to induce ferritin iron mobilization. EPO‐treatment resulted in a marked drop in the proportion of ferritin oligomers compared to the controls. To determine whether DMT1 was mediating return of iron from lysosomes to cytosol, we preloaded J774 mouse macrophage lysosomes with iron dextran, and monitored the effects of a specific DMT1 inhibitor on formation of cytosolic ferritin. DMT1 inhibition lowered ferritin production. In mice with a conditional knockout of DMT1 in hepatocytes (provided by Mitchell Knutson), stimulation of erythropoiesis by bleeding mobilized less iron from liver, but not from spleen. We conclude that DMT1 is important for the lysosomal mobilization of ferritin iron, and that ferritin oligomers may preferentially enter lysosomes during autophagy.
Aeroallergens Derived from Alternaria alternata Stimulate COX2‐dependent Expression of Antioxidant Defense Enzymes in Human Bronchial Epithelial Cells
Authors: Patil Nandadevi , Maniak Peter , Kita Hirohito , O'Grady Scott
Published: June 2021

Alternaria aeroallergens stimulate allergic inflammation and exacerbation of respiratory distress in patients with asthma. Our previous studies have demonstrated that release of ATP and subsequent increases in intracellular [Ca2+] was the result of Alternaria induced oxidative stress. Stimulation of Human Bronchial Epithelial Cells (HBECs) with Alternaria extract for 12 hours induced the expression of several phase II antioxidant enzymes, including cyclooxygenase 2 (COX2) which exhibited a 19 fold increase in mRNA expression. HBE cells loaded with CellRox™ Orange, a fluorogenic probe used for detecting oxidative stress, revealed that blocking COX2 inhibited resveratrol mediated inhibition of Alternaria induced oxidative stress. Furthermore induction of COX2 following Alternaria exposure was associated with an increase in expression of prostaglandin D2 synthase and PGD2 secretion into the extracellular media. Moreover, 15‐deoxy Δ12,14‐protaglandin J2 (15d‐PGJ2) a metabolic product of PGD2 was also shown to decrease Alternaria induced oxidative stress. These results suggest that upregulation of COX2 following sustained exposure to Alternaria induces PGD2 synthesis and subsequent formation of 15d‐PGJ2, a known activator of Nrf2 signaling and ultimately, increased expression of phase II antioxidant enzymes.
Mice Deficient in the Src Family Tyrosine Kinase yes are Protected from Hyperoxia‐Induced Lung Injury
Authors: Nelin Leif , Jin Yi , Liu Yusen , Chicoine Louis , Chen Bernadette
Published: June 2021

Endothelial cell apoptosis is an early event in the development of inflammatory lung diseases such as acute respiratory distress syndrome (ARDS) and bronchopulmonary dysplasia (BPD). In pulmonary endothelial cell culture studies we found that the Src family tyrosine kinase (STK) yes activated caspase 3, while the STK fyn inhibited caspase 3 activation. Therefore, we tested the hypothesis that mice deficient in yes will be protected from hyperoxia‐induced lung injury. Wild‐type (WT), yes‐/‐ and fyn‐/‐ mice were exposed to >95% O2 or room air for 48 hours. The animals were weighed and lung tissue harvested. The left lungs were used for determination of wet‐to‐dry weight ratios (W/D) and the right lungs for western blotting for p21 and cleaved caspase‐3, as well as qPCR for cationic amino acid transporter (CAT)‐1, CAT‐2, arginase1, arginase2, inducible nitric oxide synthase (iNOS), and cyclooxygenase‐2 (COX2). In the mice exposed to hyperoxia, yes‐/‐ mice lost less body weight than did the WT or fyn‐/‐ mice. Furthermore, yes‐/‐mice also had significantly lower lung W/D than did the WT or fyn‐/‐ mice. Cleaved caspase‐3 protein levels were lower in the yes‐/‐mice than in WT and fyn‐/‐ mice, while fyn‐/‐ mice showed higher p21 protein levels than did WT and yes‐/‐mice. Following hyperoxia exposure the mRNA levels for CAT‐1, arginase 2, and iNOS were lower in the yes‐/‐mice than in WT and fyn‐/‐ mice. Our results demonstrate that yes is an important positive regulator of hyperoxia‐induced iNOS expression and apoptosis in a mouse model of inflammatory lung disease.
Inhibition of phosphodiesterase‐4 (PDE4) by roflumilast decreases ethanol consumption
Authors: Zhang Han‐Ting , Liu Xin , Hao Pi‐da , Yang Ming‐feng , Li Da‐wei , Zhang Zong‐yong , Sun Bao‐liang
Published: June 2021

Excessive alcohol drinking normally causes alcoholism, which is one of the most damaging psychiatric disorders in the world. However, there are no ideal treatments for alcoholism in clinic. Phosphodiesterase‐4, an enzyme that specifically hydrolyzes intracellular cyclic AMP (cAMP), may play an important role in the regulation of ethanol consumption. This is supported by the findings that inhibition of PDE4 by rolipram, a prototypic PDE4 inhibitor, reduces ethanol intake and self‐administration. Roflumilast, another selective PDE4 inhibitor, has been approved for treatment of chronic obstructive pulmonary diseases in clinic. It was of interest to know whether roflumilast altered ethanol consumption. The two‐bottle choice paradigm was used to assess ethanol intake and preference in C57BL/6J mice treated with roflumilast (1, 3, or 10 mg/kg) or rolipram (0.5 mg/kg; positive control). The effect of roflumilast was verified using ethanol drinking‐in‐dark. Locomotor activity was determined using the open‐field test. Roflumilast decreased ethanol intake and preference in two‐bottle choice in a dose‐dependent manner, with the significant change at the highest dose (10 mg/kg) of roflumilast, similar to rolipram. Neither roflumilast nor rolipram affected sucrose or quinine drinking, although roflumilast at the highest dose decreased locomotor activity. These data provide additional demonstration for the role of PDE4 in ethanol intake and suggest that roflumilast may be beneficial for treatment of alcoholism.
Cocaine‐Induced Lipidomic Alterations in Human Blood for the Study of Addictive Behaviors
Authors: Pati S. , Sahin S. , Wagner J.J. , Furnari M. , Phillips K.A. , Epstein D.H. , Preston K.L. , Cummings B S
Published: June 2021

While a majority of cocaine research focuses on identification of therapies targeting dependence, few detection strategies exist to aid in clinical determination of susceptibility or likelihood of relapse. To enhance our understanding of the full spectrum of cocaine‐induced effects, we used a lipidomics approach to investigate the ability of cocaine to remodel the blood lipidome. Lipids were extracted from 190 human blood samples (123 drug users, 67 non‐users) and analyzed using electrospray ionization‐mass spectrometry. The resulting data were assessed using multivariate analysis, which indicated differences in the lipid profiles of drug users compared to non‐users. Identification of m/z features suggested alterations in glycerolipids, fatty acids, phosphatidylcholines, and phosphatidic acids. A subset of repeat abusers were asked to rate the positive and negative effects following their first cocaine exposure in comparison to their most recent use. These scores were used to assess the extent of enhanced rewarding and/or adverse effects of the drug, referred to as “sensitization”. Relative levels of several m/z features found to be significantly altered between the lipid profiles of sensitized and nonsensitized cocaine users correlated with positive and/or negative sensitization scores. These findings indicate a potential effect of cocaine on the blood, which could be exploited for the identification of lipid species correlating to the occurrence of cocaine‐induced responses and addictive behaviors in humans.

*This work was sponsored in part by the National Institute on Drug Abuse (NIDA), Intramural Research Program, National Institutes of Health (NIH).
FGF21 Mediates Alcohol‐Induced Adipose Tissue Lipolysis by Activation of Systemic Release of Catecholamine in Mice
Authors: Zhao Cuiqing , Liu Liming , Mohammadi Moosa , McClain Craig , Feng Wenke
Published: June 2021

Alcohol consumption leads to adipose tissue lipoatrophy and mobilization of free fatty acids (FFA), which contributes to hepatic fat accumulation in alcoholic liver disease (ALD). Fibroblast growth factor 21 (FGF21) is a metabolic regulator and plays an important role in glucose and lipid homeostasis in various diseases. Here, we demonstrate that FGF21 knockout (KO) mice are resistant to chronic‐binge alcohol exposure‐induced epididymal white adipose tissue lipolysis. Alcohol consumption increases FGF21 expression both in the liver and adipose tissue. Alcohol exposure causes adipose intracellular cAMP elevation, and lipolytic enzymes and coating protein activation, leading to FFA mobilization in both wild type and FGF21 KO mice. However, alcohol‐induced systemic elevation of catecholamine release, which is known to be a major player in adipose lipolysis by binding to the β‐adrenergic receptor and subsequently PKA activation, was markedly inhibited in KO mice. Supplementation with recombinant human FGF21 to alcohol‐exposed KO mice caused increased fat loss in parallel with an increase of circulating norepinephrine concentration. Furthermore, alcohol consumption‐induced fatty liver is blunted in the KO mice, indicating an inhibition of fatty acid reverse transport from adipose to the liver in the KO mice. In conclusion: FGF21 deficient mice are protected from alcohol‐induced adipose tissue excess‐lipolysis through a mechanism involving sympathetic nervous system (SNS) activation and catecholamine release. Strategies targeting the SNS activation to block FGF21‐mediated excess‐adipose lipolysis might be developed to prevent/treat alcohol‐induced fatty liver disease.
Discovery and Characterization of Novel BKCa Channel Modulators Using a New Cell‐Based Assay Platform
Authors: Lee Sojung , Lee Byoung‐Cheol , Park Chul‐Seung
Published: June 2021

The large‐conductance calcium‐activated potassium channel (BKCa channel) is expressed in various tissues and considered as an attractive therapeutic target due to its critical physiological functions. We recently developed a cell‐based assay system utilizing a hyperactive mutant BKCa channel (J Biotechnol. 167:41‐6, 2013). Since an additional maneuver to raise intracellular Ca2+ is not necessary in this assay, the activity of the mutant BKCa channels can be monitored using fluorescence measurement under physiological membrane voltages. Using the cell‐based assay, we have screened approximately 2,000 compounds of synthetic and natural origins, and identified initial candidate compounds of 13 activators and 12 inhibitors. We then investigated the mechanism of action for a single natural compound using electrophysiological means. This compound potently activated the BKCa channel from the outside of the cells by shifting the voltage‐dependent activation profile to more negative voltages. In the presence of the compound, the deactivation of macroscopic channel current was dramatically slowed down suggesting that the binding of the activator stabilizes the channel's open conformation. We are currently examining the characteristic of BKCa channel activator at a single‐channel level. Our findings verify that the newly developed cell‐based assay can be effectively applied for high‐throughput screening of BKCa channel and help to discover new sets of the channel modulators.
Synthetic Membrane Anchored Ligands: An Innovative Approach to Drug Discovery
Authors: Harwood Benjamin , Doyle Jamie , Fortin Jean‐Philippe , Raman Venkata , Krishnaji Subrahmanian , Kumar Krishna , Kopin Alan
Published: June 2021

Novel strategies are needed to expedite the generation and optimization of peptide probes targeting G protein‐coupled receptors (GPCRs). We have previously shown that membrane tethered ligands (MTLs), constructs comprised of a membrane anchor, an extracellular linker, and a peptide ligand, can be used to develop targeted receptor modulators. Although MTLs provide a useful tool to identify and/or modify active peptides, a major limitation of this strategy is the reliance on recombinant protein expression. We now report the generation and pharmacological characterization of prototype peptide‐linker‐lipid conjugates, synthetic membrane anchored ligands (SMALs), which are designed as mimics of corresponding recombinant MTLs. Using a range of GPCRs (NK1R, CCK2R, CMKLR1, MrgprX1, GLP‐1R), we illustrate the feasibility of the approach and highlight selected pharmacological properties of MTLs/ SMALs both in vitro and in vivo. Our results suggest that MTLs offer an expedited method to validate peptides that will have enhanced function as SMALs. Once identified and optimized as an active MTL, SMAL mimics can be generated and characterized. These synthetic constructs offer the possibility of direct administration rather than recombinant expression. Our work illustrates a two‐step approach of optimizing MTLs and then synthesizing corresponding SMALs. These membrane anchored probes provide powerful tools to investigate GPCR physiology and provides a basis for the development of novel therapeutics. Efforts are in progress to apply the MTL/ SMAL approach to the study of other cell surface proteins.