bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of high protein diets and leucine supplementation on body weight regulation and energy metabolism in mice
Authors: Noatsch Anne , Petzke Klaus J. , Klaus Susanne
Published: June 2021

High dietary protein (P) intake can improve glucose homeostasis, decrease body weight gain, and increase energy expenditure (EE). Leucine (Leu) was suggested to be involved in the regulation of genes stimulating energy metabolism. Therefore, we aimed to study the effects of high P and a high Leu containing adipogenic diets in mice. Male C57BL/6 mice were fed for 20 wks high fat diets (20%) containing either 10% (AP, control) or 50% whey P (HP). A third group was exposed to AP supplemented with L‐Leu (AP+L) as contained in HP. HP feeding and Leu supplementation resulted in significantly (P<0.05) lower food intake, reduced body weight gain (20.7±0.8, 14.8±1.4 and 9.7±0.8 g for AP, AP+L and HP, respectively), lower food energy efficiency (2.14±0.08, 1.57±0.11 and 1.11±0.08 g/MJ for AP, AP+L and HP, respectively), lower body fat mass (quantitative nuclear resonance), lower liver triacylglycerol concentrations, and higher lean body mass as well as higher weights of m. quadriceps and higher postabsorptive 15N‐lysine incorporation into skeletal muscle. EE (indirect calorimetry) was not different between groups. Parameters of glucose homeostasis were significantly (P<0.05) improved in HP and AP+L fed mice. Effects of Leu supplementation are generally intermediate to those of the HP as compared to AP. We suggest a role of Leu at the increase of EE; however, an expected higher uncoupling protein gene expression was not detected.
Effects of dihydrocapsiate on diet‐induced thermogenesis following 4 weeks of very low calorie dieting.
Authors: Lee T.Y. Amy , Zerlin Alona , Thames Gail , Li Zhaoping , Heber David
Published: June 2021

Dihydrocapsiate (DCT) is a naturally occurring compound which is structurally related to capsaicin from hot pepper but found in a non‐pungent pepper CH‐19 Sweet. It is also a well‐studied potential food ingredient that elicits some of favorable metabolic effects experienced from capsaicin without side effects. In order to determine whether DCT increased post‐prandial energy expenditure (PPEE) in response to a 400 Cal/60 g protein liquid test meal high (Pro‐Cal 100, R‐Kane, NJ) following a very low calorie diet (800 Cal and 120 g per day) utilizing the same liquid protein meals prescribed for 4 weeks, 51 subjects were entered into a randomized prospective controlled study and were given two doses of DCT (Ajinomoto USA, NJ) as capsules three times per day (3mg or 9 mg) or placebo. Data for 33 subjects (n=9 placebo; n=11@ 3 mg; n=13@ 9 mg) completing the study were evaluated. At baseline and 4 weeks, fasting basal metabolic rate and PPEE were measured in a metabolic hood (Vmax, Sensormedics). Fat free mass (FFM) was determined using a Bod Pod. Expected decreases in adaptive thermogenesis were not affected by DCT administration. PPEE normalized to FFM was increased significantly with peak effect at 1 hour after meal administration in the group receiving 9 mg/day by comparison to placebo (p<0.05). Respiratory quotient (RQ) increased by 0.04 in the placebo group (p<0.05) after the high protein meal, but did not change in groups receiving DCT. DCT resulted in increased PPEE after meals and inhibition of increase in RQ after meals during 4 weeks of very low calorie dieting suggesting it may provide metabolic enhancement to weight management efforts.
Maternal fructose consumption programs gene expression pattern in intestine of male offspring
Authors: Arentson Emily , Zou Mi , Teegarden Dorothy , Buhman Kimberly K , Donkin Shawn S.
Published: June 2021

Metabolic programming of insulin‐sensitive processes by maternal diet has been demonstrated for liver, pancreas, and muscle of offspring, but has not been evaluated for small intestine. To determine the effects of maternal (MAT) and postweaning diet (PWT) on intestinal programming, male offspring of Sprague Dawley females fed either a 60% starch diet (CON), 60% fructose diet (FRU) or protein restricted diet (LP) throughout pregnancy and lactation were weaned to either a CON, FRU or a 30% fat diet (FAT). Intestinal mRNA expression was evaluated at 18 weeks for all nine MAT×PWT groups. Glucose 6 phosphatase (G6Pase) was elevated in FRUFAT compared with FRUCON offspring (3.98 ± 0.70 vs. 1.16 ± 0.53, P < 0.05) with a similar tendency for PEP carboxykinase (PCK1) (2.95 ± 0.62 vs. 1.53 ± 0.47, P=0.07). G6Pase and PCK1 mRNA levels in offspring from CON and LP dams were not affected by the FAT diet (G6Pase: CONFAT vs. CONCON, 0.46 ± 0.50 vs. 1.17 ± 0.64, LPFAT vs. LPCON, 0.84 ± 0.56 vs. 1.92 ± 0.56, PCK1: CONFAT vs. CONCON, 0.57 ± 0.44 vs. 1.33 ± 0.53, LPFAT vs. LPCON, 0.95 ± 0.53 vs. 0.82 ± 0.53). Collectively, offspring from FRU dams had more (P < 0.05) PCG‐1α mRNA compared with offspring of CON or LP dams (1.31 ± 0.14 vs. 0.89 ± 0.13 or 0.85 ± 0.14, respectively). These data indicate a persistent adaptive response for intestinal gene expression in offspring that is sensitive to maternal fructose consumption and amplified with postweaning fat intake.

Grant Funding Source: NIH DK077581
UPREGULATION OF PI3K‐AKT PATHWAY FOLLOWING TREATMENT WITH EGFR SPECIFIC shRNA IN REGENERATING RAT LIVERS
Authors: Paranjpe Shirish , Bowen William C. , George Tseng C. , Luo Jian‐Hua , Orr Ann V. , Michalopoulos George K.
Published: June 2021

Liver regeneration after two‐third partial hepatectomy (PHx) is a complex process requiring interaction and cooperation of many growth factors and cytokines and crosstalk between multiple pathways. To investigate the role of EGFR in liver regeneration, we used two EGFR specific shRNA to inhibit EGFR expression in regenerating normal rat liver. Suppression of EGFR mRNA and protein was evident in treated rats. There was also a demonstrable decrease but not complete elimination of BrdU incorporation and mitoses at 24 hours after PHx, proapoptotic genes were upregulated, and genes associated with cellular proliferation were down regulated. However a compensatory upregulation in ErbB‐3, ErbB‐2, MET and Src resulted in an increase in formation of ErbB‐3‐ErbB‐2 heterodimers which in turn activated the PI3K‐Akt survival pathway and restoration of liver mass by hepatic hypertrophy. However, we did not observe any mortality associated with silencing of EGFR and no delay in regeneration was observed in EGFR silenced rats. The data indicate that although the MET and EGFR pathways are similar, the contributions made by MET and EGFR are unique and are not compensated by each other or other cytokines.
Blood‐brain barrier injury during spirochetal infection
Authors: Londono Diana , Cadavid Diego
Published: June 2021

The most severe and a relatively common complication of untreated relapsing fever (RF) borreliosis is brain hemorrhage; mortality rate can reach as high as 70% in the epidemic form. Subjects with epidemic RF who are stable prior to treatment have extraordinarily high levels of circulating IL‐10. Our studies in murine RF borreliosis have shown that IL‐10 protects the microcirculation of the brain and others organs from hemorrhagic and thrombotic complications. Mortality is higher in infected mice deficient only in IL‐10 compared to mice normal for IL‐10 but deficient in both B and T cells. In this study we examined morphologically and molecularly the brain of mice with relapsing‐remitting and persistent infection with B. turicatae (Bt) with or without IL‐10 deficiency for alterations in the BBB. The analysis shows significant BBB disruption in IL‐10 deficient infected mice often resulting in brain hemorrhage. Immufluorescence microscopy of vessels near hemorrhagic areas shows rupture of basal lamina, detachment of astrocytes foot processes and pericytes, leakage of serum proteins, decreased in tight junction proteins, endothelial cell activation and apoptosis, and recruitment of inflammatory cells. These results indicate that IL‐10 protects the BBB at multiple levels during spirochetal infection to prevent injury. Studies funded by NIH grant R21 NS057545‐02 to Diego Cadavid.
Regional localization of intestinal dendritic cell subsets control Th‐17 responses
Authors: Denning Tim , Norris Brian , Greene Brent , Pulendran Bali
Published: June 2021

Mucosal immunity is influenced in large part by the dynamic interplay between various intestinal antigen‐presenting cell subsets and T lymphocytes. Using 10‐color flow cytometry, we have defined the major DC and macrophage subsets in the small and large intestine LP as distinguishable by the reciprocal expression of CD103 and F4/80, respectively, and not exclusively by CD11b or CD11c. Intestinal F4/80+ macrophages expressed IL‐10, TGF‐β, retinoic acid‐generating enzymes and promoted the differentiation of inducible FoxP3+ regulatory T cells. Alternatively, CD11b+, but not CD11b−, CD103+ LP DCs expressed IL‐6 and TGF‐β as well as retinoic acid‐generating enzymes and efficiently induced the differentiation of TH‐17 cells. These CD103+ LP DC subsets displayed striking region‐specific localization along the intestine with CD11b−CD103+ LP DCs appearing most abundantly in the large intestine while CD11b+CD103+ LP DCs were most prevalent in the upper small intestine. The distribution pattern of CD11b+CD103+ LP DCs in particular correlated with the representation of TH‐17 cells in the intestine and during intestinal inflammation CD11b+CD103+ LP DCs accumulated in the colonic LP as did TH‐17 cells. Our results suggest that CD11b+CD103+ LP DCs may be targets for modulating IL‐17 producing T cell responses during intestinal inflammation.
Voltage‐dependent K+ channels in Immunogenic and Regulatory Functions of Lung Dendritic Cells
Authors: Wang Xiang , Shao Zhifei , Agrawal Devendra K.
Published: June 2021

Dendritic cell (DC) subsets display different functional roles in regulating immune responses. We identified two DC populations, CD11clowCD11bhigh and CD11chighCD11blow, in the lungs of asthmatic mice. The DC subsets show distinct expression patterns in their surface markers and chemokine receptors. Since K+ channels modulate the function of T‐lymphocytes and mast cells, we examined ion channel expression of both DC subsets using whole‐cell voltage‐clamp techniques. At least two types of voltage‐dependent K+ currents were detected in both DCs, demonstrated by differential inhibitory effects of the selective Kv1.3 blocker, margatoxin, and the non‐selective voltage‐sensitive Ca2+‐activated K+ channel blocker, charybdotoxin. Voltage‐dependent proton currents were also detected in both DCs, and this was blocked by 10 mM Zn+. This suggests the potential role of lung DCs in the phagocytosis through activating the NADPH oxidase. The CD11clowCD11bhigh DCs exhibited significantly larger amplitudes in all current than CD11chighCD11blow DCs (p<0.01), suggesting that CD11clowCD11bhigh DCs possess a higher ion channel expression level in the membrane. This differential expression level of ion channels may shed light on the distinct functional properties between the two DC subsets and their migration to the lymph node during allergic immune response.

(Supported by LB506 DHHS State of Nebraska cancer and Smoking‐related Disease Program)
Matrix tethered EGF promotes survival of MSC secondary to preferential activation of surface‐restricted signaling pathways
Authors: Rodrigues Melanie , Sanchez Edgar , Griffith Linda , Wells Alan
Published: June 2021

Multipotential stromal cells (MSC) are being tested to replace traditional tissue transplantations. However, the major loss of MSC seen after implantation may be due to non‐specific inflammation killing these cells. Epidermal Growth Factor (EGF) promotes MSC expansion but does not cause MSC differentiation and when covalently tethered on an engineered biomaterial surface (tEGF) offers a survival advantage to MSC in the presence of FasL, while soluble EGF (sEGF) increases death. Thus, we hypothesize that tEGF by restricting signaling of EGF receptor to the plasma membrane changes the persistence and balance of intracellular signaling pathways towards survival in MSC. Our results show caspase8 and caspase3 activation upon treatment with FasL. Addition of sEGF to these cells shows enhanced apoptosis in line with earlier findings. However a key question is what occurs as MSC differentiate into the desired tissue. We now propose that as MSC differentiate their susceptibility to death signals diminish. Preliminary studies show reduced apoptosis in response to FasL as MSC are driven down the osteogenic lineage. Studies with tEGF are underway. Still, the early findings of relative resistance to death of the differentiating cells suggest that the tEGF survival signal need not persist which allows for a degradative scaffold to be used to repair wounds including non‐healing bone lesions.

Funding: NIGMS (AW), NIDCR (LG)
Passion and Technology: Separation of Egyptian Conjoined Twins
Authors: Salyer Kenneth E
Published: June 2021

Advances in technology in the last 30 years have exponentially improved accuracy in the diagnosis, treatment, and delivery of technically advanced surgery. Craniofacial surgery has become safer and more precise by using 3‐D models. These models are produced by using CT scan and MRI data which is stereographically or digitally implemented to provide anatomical reconstruction, which serves as a road map for the surgical team. Accurate pre‐operative sequencing, timing, and approaches can be planned out in detail before the surgery is performed.

Dismal results over the last 400 years have prompted some critics to condemn surgical attempts at separation of craniopagus twins. Alice Dreger says, “They shouldn't be separated; society needs to change its attitude about deformity.” Very few craniopagus twins have been successfully separated, and functioned in society.

Two Egyptian boys attached extensively at the head, shared their brain at the motor cortex. Their skulls were fused completely and shared their mutually conjoined head anatomy.

Extensive studies, including complete anatomic body models with 3‐D brain, venous system, and other vital anatomic structures, were produced from CT and MRI data. Ultimately, a complete step by step 36 hour operation was planned and executed, leading a team of 50 medical caregivers. This on‐going story will be relayed in detail during this presentation.
Advanced digital technology in prosthetic reconstruction
Authors: Verma Suzanne Nicole
Published: June 2021

Anaplastology is the art and science of restoring a malformed or absent part of the human body through artificial means. These custom prostheses are created to mimic contour, form, and color of the missing anatomy, and can provide the patient the opportunity to function in society as they would previously. Fabrication of facial prostheses has been revolutionized as digital technologies have opened the door to a substantial shift in the way anaplastologists conceptualize and provide facial prosthetic treatment. Computer‐driven systems have contributed to the exploitation of 3D imaging, simulation programs, CAD/CAM, and rapid prototyping systems, and helped identify new, accurate, cost and time‐efficient means of fabricating prostheses and providing patient treatment.
Tactile Medical Modeling Using Additive Manufacturing Technologies
Authors: Christensen Andrew M.
Published: June 2021

Three‐dimensional visualization of complex anatomical and biological structures has enabled research and clinical care to progress at rapid speed. Advances in medical imaging and physical biomodeling have allowed health care providers to prepare for complex reconstructive surgery using patient specific virtual and tactile data. Thanks to advances in the additive manufacturing field, tactile medical and biological objects of extreme complexity and accuracy can be created from medical imaging. Robust materials and multi‐material capable machines now produce models for more effective research and clinical planning.
The digital reconstruction process and the assessment of hard tissue structure and mechanics
Authors: Zapata Uriel , Wang Qian , Opperman Lynne A , Dechow Paul C
Published: June 2021

Modern scanning methods have made anatomical digital reconstruction critical for assessing skeletal function. Hard tissues from the crania of 4 different species were used to assess digital reconstruction for accurately modeling relationships between shape and function. Digital information from humans, alligators, dogs, and rhesus monkeys were collected by using computed tomography scans. DICOM files were processed by using segmentation methods to separate tissues. Once hard tissue shapes were obtained, functional conditions were assigned to the morphology, and biomechanical characteristics were considered to evaluate the functional load effects. Three‐dimensional morphologies were obtained in every case. However, acquiring tissue separation was a challenging process affected by both the precision of data collection and experience with the software. Interestingly, biomechanical features, material properties, and physiological actions were very important in modeling function. The digital reconstruction of hard tissues is the first step to understanding the relationship between function and morphological features. But the process is incomplete without a comprehensive definition of functional actions, material, and biomechanical characteristics of the anatomical elements. Supported by NSF BCS 0725126, NIH/NIDCR DE017259‐DE015437, and COLCIENCIAS (Bogotá, Colombia).

Grant Funding Source: NIDCR DE017259 ‐ DE015437
Fetal Origins of Lung Disease: PPARγ and Epigenetics
Authors: Joss‐Moore Lisa
Published: June 2021

Fetal and early life events impact the developing lung, contributing to an increased risk of bronchopulmonay dysplasia and potentially increasing the incidence of adult pulmonary disorders. The transcriptional regulator PPARγ is important for lung development and its effects are likely the result of epigenetic mechanisms. Epigenetic determinants include DNA methylation and covalent modifications of histones. PPARγ gene products arise from the use of multiple promoters and alternative splicing; as such, PPARγ relies on epigenetics to affect appropriate gene expression. In addition, recent studies have demonstrated that PPARγ targets the histone lysine methyltransferase, Setd8, an enzyme that places a methyl group on lysine (K)20 of histone(H) 4. Importantly, Setd8 places this mark on histones associated with the PPARγ gene in addition to other targets. Animal studies have demonstrated reduced PPARγ mRNA and protein levels in the lung in response to numerous perinatal insults. In fetal growth restricted rat lungs, PPARγ levels are reduced in conjunction with reduced Setd8 levels. The H4K20Me mark is also decreased on the PPARγ gene and other genes important in lung developmental outcome. We speculate that one way that PPARγ effects lung development is by regulating levels of Setd8 and the H4K20Me mark on PPARγ as well as other target genes.
Nanoformulated superoxide dismutase 1 (SOD1): Implications for angiotensin II (AngII) and brain‐related cardiovascular diseases
Authors: Rosenbaugh Erin , Roat James , Gao Lie , Yang Rui‐Fang , Manickam Devika S. , Yin Jing‐Xiang , Schultz Harold D. , Bronich Tatiana K. , Batrakova Elena V. , Kabanov Alexander V. , Zucker Irving H. , Zimmerman Matthew C.
Published: June 2021

Due to possible toxicity and the inability of adenoviral vectors to target the brain, the therapeutic potential of adenoviral‐mediated gene transfer of SOD1 for the treatment of AngII‐dependent neuro‐cardiovascular diseases is limited. Thus, we developed a nanotechnology‐driven delivery system using a polyion complex micelle with a polyethylene glycol corona and a polyethyleneimine core that electrostatically binds SOD1 to form SOD1 nanozyme. We hypothesized that SOD1 nanozyme inhibits AngII intra‐neuronal signaling by delivering functional SOD1 to neurons. In catecholaminergic (CATH.a) neurons, SOD1 nanozyme, but not free SOD1 protein, inhibited the AngII‐induced increase in superoxide, as measured by DHE fluorescence and EPR spectroscopy. The AngII‐induced inhibition of K+ current (IKv) was also attenuated (P<0.05) in SOD1 nanozyme‐treated neurons (10 ± 1% IKv inhibition) compared to control and free SOD1‐treated neurons (38–40 ± 5% IKv inhibition). In conscious rabbits, the pressor response (21 ± 2 mmHg) induced by central‐injection of AngII (100 ng) was inhibited (6 ± 2 mmHg; P<0.05 vs. control) by intracarotid (i.c.) injection of SOD1 nanozyme (400 U/mL), but not free SOD1 protein (21 ± 2 mmHg). These data suggest that nanomedicine‐based delivery of SOD1 may provide a new therapeutic strategy for AngII‐dependent cardiovascular diseases associated with increased superoxide in the brain.
Lipid transporters and binding proteins; MsbA and NPC1
Authors: Sharom Frances J.
Published: June 2021

Understanding the molecular mechanism of proteins that translocate or transfer lipids presents a special challenge. The bacterial ABC protein, MsbA, is a homodimeric “half‐transporter” of known structure, which is thought to act as an outwardly‐directed ATP‐driven flippase for lipid A and possibly some phospholipids. We have purified MsbA with high ATPase activity and established its binding affinity for putative substrates using fluorescence spectroscopic tools. Reconstituted MsbA displays a high level of flippase activity for certain phospholipids, providing an excellent opportunity to explore the mechanism of lipid translocation at the molecular level.

Mutations in NPC1 cause Niemann‐Pick Type C disease, a fatal neurodegenerative condition characterized by accumulation of cholesterol and glycolipids in the late endosomes. NPC1 is a 190 kDa integral protein found in the late endosomal membrane. It is known to be a key player in intracellular cholesterol trafficking, but the nature of its substrates and its exact role remain enigmatic. Recent studies on purified NPC1 have shown that it binds several sterols, and the binding site has been localized to the N‐terminal luminal domain, which has been crystallized in sterol‐bound form. NPC1 may accept sterol from NPC2, a small soluble protein in the lumen of late endosomes, and transfer it to either the membrane, or an as yet unidentified cytoplasmic acceptor.
Metabolism as a key to HDAC inhibition by dietary constituents
Authors: Dashwood Roderick H.
Published: June 2021

Histone deacetylase (HDAC) inhibitors are gaining interest as cancer therapeutic agents. We have pursued this avenue from the perspective of dietary constituents which, after metabolism, alter HDAC activity in cancer cells. Butyrate, a short‐chain fatty acid generated via gut fermentation of dietary fiber, was shown over 30 years ago to affect histone status in erythroleukemia cells, via competitive HDAC inhibition. Cruciferous vegetables such as broccoli contain glucobrassicin, the precursor of sulforaphane (SFN). Metabolism of SFN and other isothiocyanates via the mercapturic acid pathway generates intermediates with HDAC inhibitory activity. Allium vegetables contain various organosulfur compounds which can be converted to small molecule thiols, such as allyl mercaptan, that act as competitive HDAC inhibitors. Natural organoselenium compounds, such as Se‐methylselenocysteine (MSC), are found in selenium‐rich foods such as Brazil nuts. Glutamine transaminase K converts MSC to methylselenopyruvate (MSP), and molecular modeling supported the interaction of MSP with zinc in the HDAC pocket. In colon and prostate cancer cells, MSP was a potent HDAC inhibitor. Thus, various dietary anti‐cancer agents alter HDAC activity and histone acetylation status. Studies supported by NIH grants CA122959, CA090890, CA65525, and P30 ES00210.
EPIGENETICS AND METABOLISM: THE CIRCADIAN CLOCK CONNECTION
Authors: Sassone‐Corsi Paolo
Published: June 2021

Circadian rhythms govern a number of fundamental physiological functions in almost all organisms, from prokaryotes to humans. Disruption of these rhythms can have a profound influence to human health and has been linked to depression, insomnia, jet lag, coronary heart disease and a variety of neurodegenerative disorders. The complex program of gene expression that characterizes circadian physiology is possible through dynamic changes in chromatin transitions. A central element of the clock machinery, the protein CLOCK, has HAT enzymatic properties. It directs acetylation of histone H3 and of the non‐histonic protein BMAL1, its own dimerization partner, at K537, an event that is essential for circadian function. We show that the HDAC activity of the NAD+‐dependent SIRT1 enzyme is regulated in a circadian manner. SIRT1 associates with CLOCK and is recruited to the CLOCK:BMAL1 chromatin complex at circadian gene promoters. Genetic ablation of the Sirt1 gene or pharmacological inhibition of SIRT1 leads to disturbances in the circadian cycle. Finally, using liver‐specific SIRT1 mutant mice we show that SIRT1 contributes to circadian control in vivo. We propose that SIRT1 functions as an enzymatic rheostat of circadian function, transducing signals originated by cellular metabolites to the clock machinery. One question however arose: how acetylation can be oscillatory if the intracellular levels of CLOCK and SIRT1 do not cycle? We demonstrate that intracellular NAD+ levels cycle with a 24 h rhythm, an oscillation driven by the circadian clock. CLOCK:BMAL1 regulate the circadian expression of Nampt (nicotinamide phosphoribosyltransferase), a rate limiting step enzyme in the NAD+ salvage pathway. Our findings reveal an interlocked transcriptional‐enzymatic feedback loop that governs the interplay between cellular metabolism and circadian rhythms.
Quantitative Modeling of a Transcriptional Grammar: Short‐Range Repression in Drosophila
Authors: Arnosti David N. , Ay Ahmet , Fakhouri Walid , Sayal Rupinder , Dresch Jacqueline , Clifford Jacob , Chiu Chichia
Published: June 2021

Developmental gene regulation is characterized by complex networks of transcription factors and enhancers, but analysis of cis regulatory information on a genomic level by direct experimentation remains a challenge. Quantitative modeling offers an alternative path to develop a global understanding of the transcriptional regulatory code. Recent studies have focused on endogenous regulatory sequences; however distinct enhancers differ in many features, making it difficult to generalize to other cis‐regulatory elements. To isolate the effects of factor spacing, stoichiometry, and arrangement, we applied a systematic, quantitative approach to 27 simpler regulatory elements that were analyzed in the context of the Drosophila blastoderm embryo. Using a database of almost 1000 images, we present here the first quantitative analysis of short‐range transcriptional repressors, which play central roles in metazoan development. Our fractional occupancy‐ based modeling uncovered unexpected features of these proteins' activity that allow accurate predictions of regulation by the Giant, Knirps, Krüppel and Snail repressors, including modeling of endogenous enhancer sequences. This study provides essential elements of a transcriptional grammar that will allow extensive analysis of genomic information in Drosophila melanogaster and related organisms.
Increased expression of hPygo2 is a cellular response to HPV infection in cervical dysplasia
Authors: Popadiuk Cathy , Tzenov Youlian , Andrews Phillip , Kao Kenneth R
Published: June 2021

We previously demonstrated that the Wnt‐signaling component, hPygo2 was required for cancer growth and that ELF‐1, which is normally regulated by the retinoblastoma (Rb) tumor suppressor, activates hPygo2 expression. Because the E7 protein of Human Papillomavirus (HPV) suppresses Rb function, we hypothesized that hPygo2 expression would be persistent in cervical cells transformed by HPV. Western and immunofluorecence analyses indicated that primary endocervical cell lines (HEN) did not express hPygo2, but that HPV‐transformed endocervical cells and cervical cancer cell lines highly expressed hPygo2. Transfection of a dominant active form of Rb into HPV infected cells reduced hPygo2 expression. Immunohistochemical analysis of hPygo2 and HPV protein expression in a tissue microarray of cervical cancer progression showed weak expression in nuclei of parabasal cells of normal ectocervical epithelium. CIN II and III staged dysplasias showed high levels of expression in cytoplasm and the highest levels of expression were found in squamous cell carcinomas, primarily in the cytoplasm. HPV antibodies stained nuclei and cytoplasm of non‐neoplastic tissues very strongly, and to a lesser extent CINII, III and squamous cell carcinomas. These observations suggested that hPygo2 protein accumulates increasingly with dysplasia leading to frank cancer.
NPM‐ALK signals through glycogen synthase kinase 3β to promote oncogenesis
Authors: McDonnell Scott Robert Paul , Wey Elizabeth , Cheng Mengang , Zeng Zihua , Zu Youli , Elenitoba‐Johnson Kojo , Lim Megan
Published: June 2021

Anaplastic large cell lymphoma (ALCL) is characterized by the expression of nucleophosmin‐anaplastic lymphoma kinase (NPM‐ALK). The mechanism of activation of downstream molecules that regulate cell growth in ALCLs has not been elucidated. Using mass spectrometry‐based phosphoproteomic screen, we identified GSK3β as a potential signaling protein of NPM‐ALK. We hypothesize that NPM‐ALK signals through GSK3β and contributes to the oncogenic phenotype of ALCL. To assess the role of GSK3β in NPM‐ALK induced oncogenesis, we utlilized an ALK inhibitor and stable tetracyclin‐inducible shRNA knockdown of NPM‐ALK in ALCL‐derived cell lines. ALCL derived cell lines were evaluated for cell viability and levels of P‐S9GSK3β and NPM‐ALK protein. The expression of P‐S9GSK3β and CDC25A (a known GSK3β substrate) was evaluated in biopsies of ALCL tissues using immunohistochemistry. Inhibition of ALK kinase activity resulted in a dose‐dependent decrease in cell viability which was rescued by a small molecule inhibitor of GSK3β (GSK3 IX). NPM‐ALK kinase activity was critical for the phosphorylation of GSK3β at serine‐9 but not total GSK3β. P‐S9GSK3β and CDC25A were expressed in neoplastic cells of ALCL tissue biopsies. Our results provide support for the role of GSK3β/CDC25A as mediators of NPM‐ALK oncogenesis. Funding support comes from University of Michigan, Department of Pathology development funds.
Altered TLR2 Expression and Signaling contribute to the Hyperinflammation in diabetic wounds
Authors: Dasu Ravi Krishna Mohan , Jialal Ishwarlal
Published: June 2021

Toll‐like receptor‐2 (TLR2) is a pivotal pathogen recognition signaling receptor and plays a key role in inflammation and injury. Hyperglycemia, inflammation, and oxidative stress induce TLR2, and are major pathophysiological mechanisms in impaired diabetic wound healing process. However, an in vivo contribution of TLR2 to the inflammatory state of diabetic wounds remains elusive. The aim of the study is to examine the contribution of TLR2 to the hyperinflammation in wounds of diabetic mice. Diabetes was induced in male C57BL/6J (WT) and TLR2 Knockout mice (KO) using streptozotocin with suitable non diabetic controls (n=6/group). Full thickness excisional wounds were made on the backs of all the mice aseptically. Total RNA and protein were subjected to RT‐PCR, Western blots, and wound sizes were recorded. TLR2 mRNA expression increased 2.5 fold in diabetic wounds of WT mice (P<0.05) with corresponding Myd88 expression, IRAK‐1phosphroylation, and NF‐κB activation compared to non diabetic wounds. Wounds of diabetic TLR2KO mice showed less lipid peroxidation (−70%), Myd88 signaling, NF‐κB(−62%), IL‐1β(−82%), and smaller wound sizes compared to diabetic WT mice after 10 days. Collectively, our findings demonstrate that increased TLR2 expression and signaling contribute to the diabetic wound hyper inflammatory state and TLR2 deficiency decreases inflammation and improves diabetic wound healing.
Immunogenicity of Avian H5N1 Influenza Virus Recombinant Vaccines in Cats
Authors: Uhl Elizabeth W , Tompkins S. Mark , Harvey Stephen B , Gabbard Jon , Michel Frank , Perozo Yaneth , Hogan Robert J
Published: June 2021

Domesticated cats can be infected with highly pathogenic avian influenza H5N1 virus and potentially can be vector or reservoir hosts in an influenza pandemic. We examined the immunogenicity of the hemagglutinin (HA) of H5N1strain VN/1203/04 in cats. ELISAs showed that a single dose of recombinant H5 HA protein can induce a robust antibody response against both whole inactivated virus and recombinant HA antigen. A marked increase in hemagglutination inhibition (HI) titers was also observed in sera from cats given a single dose of the H5 HA protein. However, cats given the commercial H5N1 vaccine required 2 doses before increased H5 HA‐specific antibody titers were observed. Despite boosting, cats receiving the commercial H5N1 vaccine had HI titers lower than those given one injection of recombinant protein. Cats vaccinated with plasmid DNA encoding HA failed to develop HA‐specific antibody responses. Finally, sera from cats that received the protein vaccine neutralized wild type H5N1 influenza virus (VN/120304). The results of this study indicate that recombinant H5 HA protein‐based vaccines may be more effective that either inactivated influenza virus or DNA vaccines in cats.
Proton pump inhibitor modulation of gastric mucosal CINC‐1 in indomethacin‐induced rat gastritis
Authors: Zinkievich J. Michael , Nandi Jyotirmoy , Acharya Nisha , Jha Shalinee , Levine Robert A.
Published: June 2021

Lansoprazole, a proton pump inhibitor, improves indomethacin‐induced gastritis by decreasing gastric acid secretion. Rat CINC‐1 is the homologue to the human growth‐regulated oncogene product, is increased at sites of inflammation, and induces neutrophil influx. Our aim was to investigate the modulation of inflammatory chemokines by lansoprazole in indomethacin‐induced rat gastritis. Male Sprague‐Dawley rats (n = 6/group) were treated orally with indomethacin (25mg/kg), with or without lansoprazole (10mg/kg) pretreatment. Control rats received vehicle alone. Rats were euthanized 4 h or 18 h after drug or vehicle administration. Ulcer index (UI), myeloperoxidase (MPO) activity, malondialdehyde (MDA), prostaglandin E2 (PGE2), and CINC‐1 were measured. Compared to control, indomethacin significantly increased UI (470%), mucosal MPO activity (250%), MDA level (267%), and tissue CINC‐1 (395%) at 4 h. These values remained elevated at 18 h. PGE2 synthesis was markedly inhibited by 85% with indomethacin at 4 h, but rebounded to near control levels at 18 h. Serum CINC‐1 was unchanged at 4 h, but was elevated 9‐fold at 18 h with indomethacin. Pretreatment with lansoprazole significantly reduced indomethacin‐induced UI, MPO, MDA, and tissue CINC‐1 at both time points. However, lansoprazole had no effect on PGE2 synthesis or serum CINC‐1 levels. In conclusion, our data suggest that lansoprazole limits neutrophil influx by reducing mucosal CINC‐1 levels and therefore facilitates improvement of indomethacin‐induced gastritis.
Teaching pathophysiology in a systems‐based curriculum: use of a Student Auscultation Manikin (SAM)
Authors: Fitzovich Douglas E , Yonts R. Christopher
Published: June 2021

One purpose of a systems‐based preclinical medical education curriculum is to facilitate integration of basic and clinical science knowledge and skills. At this institution (LMU‐DCOM) introduction of preclinical medical students to fundamental clinical skills begins immediately. The development of activities that promote integration of basic and clinical sciences is essential for a successful systems‐based medical education. At LMU‐DCOM the use of an auscultation manikin (Cardionics) has been introduced in the second year cardiovascular systems course. The manikin permits an instructor to demonstrate the proper use of auscultation techniques as the students listen to typical normal and abnormal sounds through wireless stethoscopes. Questionnaires using a 4‐point Likert scale assessed student attitudes regarding the laboratory. Open‐ended questions regarding the cardiovascular course as a whole and the SAM were also administered. Students overwhelmingly responded positively to the SAM exercise. Responses were 68.3% positive regarding understanding of normal cardiac valvular function, 68.3% positive regarding diagnosis of valvular diseases, and 65.85% positive for recognition of non‐valvular defects. 90.2% of students wanted more time with the SAM. This laboratory was most often cited as the most effective component of the course. Performance on a written quiz immediately after the laboratory was 87.6% ± 22.4 (SD) versus 83.7% ± 6.2 for the cardiovascular system course examinations. This experience with the auscultation manikin demonstrates that simulation exercises can stimulate interest in learning and effectively promote integration of basic pathophysiology and clinical science practical procedures in the preclinical curriculum. This work was supported by LMU‐DCOM.
Interdisciplinary Physiology: Walkable Neighborhoods, Obesity & Diabetes
Authors: McFarland Jenny Lee
Published: June 2021

As science educators, one of our goals is to help students learn to solve complex problems that they will encounter as individuals, scientists and health care providers. In physiology these problems involve human health and disease; solving them requires an interdisciplinary approach inextricably linked to social and environmental factors. Students need help putting these pieces together, making connections amongst disciplines. An integrative learning activity was used in a course for liberal arts students (biology of human disease) and a sophomore level human anatomy and physiology course for science and pre‐nursing students. In this activity students explore the relationships among the physical environment, diet, exercise, obesity and diabetes as they analyze the causes and consequences of obesity and explore solutions to this complex public health issue. This activity helps students realize that, although physiological knowledge is essential for understanding the obesity epidemic, it is not sufficient. Sociological factors must also be considered as well as environmental ones. Interdisciplinary activities and explicit fostering of interdisciplinary thinking do impact student learning of physiology and student engagement in these courses.
Playing a Jeopardy‐like quiz game to integrate new material in A & P
Authors: Halpin Patricia A.
Published: June 2021

Anatomy & Physiology (A&P) is taught nationwide to allied health majors who use a wide variety of learning styles to learn the material. Many students are challenged to rapidly learn new material and at the same time to integrate the new material with their growing knowledge base. In an effort to solve this problem the class plays a Jeopardy‐like quiz game at the end of each unit of new material. The class of 30 is divided into six equal groups. Each group gets the opportunity to answer one question each round. They discuss the question and answer as a group. The winning team each week is given credit that is added to their individual grades. Using a Likert scale the class was surveyed and it was determined that 83% strongly agreed (5) that playing the game helped them review the new material. It was also determined that playing the game was a good way to integrate old material into the material of that day's lecture (39% strongly agreed (5) + 52% agreed (4)). Another benefit of the game is that the students felt it was a good way to review material for exams (26% strongly agreed (5) + 39% agreed (4)). When asked if they liked playing the game most of the class responded favorably (35% strongly agreed (5) + 43% agreed (4)). In conclusion playing the game provides a fun learning tool for students to review and integrate new material.
Frontiers in Physiology: A fellowship program that fosters the teaching and learning of science
Authors: Limson Mel , Ngo Kathleen Kelly , Matyas Marsha Lakes
Published: June 2021

The APS Frontiers in Physiology Professional Development Fellowship supports middle/high school science teachers in learning about scientific research and creating student‐centered learning environments. Fellows in the year‐long program examine their current teaching methods and learning environments, engage in cutting‐edge biomedical research alongside APS members, attend a week‐long workshop with mentor teachers and physiologists to learn effective teaching strategies and physiology content, enhance and develop classroom materials, and participate in Experimental Biology. 2006–2008 Fellows (n=45) provided qualitative and quantitative evaluation feedback via surveys (pre/post), online discussions, reflections, and portfolios. Overall, Fellows increased their understanding of the research process and how scientific knowledge is generated and shared. They also diversified and deepened their science teaching skills. The multiple components of the fellowship program demonstrate an effective strategy in fostering an integrative approach to learning both physiology content and the teaching of science. Frontiers in Physiology is sponsored by the APS, a Science Education Partnership Award from the National Center for Research Resources and the National Institute of Diabetes and Digestive and Kidney Diseases at the NIH.
Collaboration in the Classroom: Integrating Fundamental Physiological Concepts through Experimental Design and Disease
Authors: Johnson Kathryn M.S.
Published: June 2021

With the primary goal of developing “physiology intuition” by understanding how physiological systems interact, students in Human Anatomy & Physiology, an upper‐level undergraduate one‐semester course taught at a liberal arts college, paired to become a team of experts on a specific physiological system (ex. cardiovascular). Modeling how scientists often learn new content by integrating information with previous expertise, expert teams explored how larger physiological concepts, such as homeostasis, applied to their system. Mirroring collaborations in the scientific community, teams worked with other expert teams to develop a hypothesis focused on the interaction of their physiological systems. Students designed an experiment to test their hypothesis, generated simulated data, and drew conclusions based on primary literature. This laid the foundation for novel large‐scale physiological experiments designed and conducted by the students. Later in the semester, partnerships explored diseases that affect both of their physiological systems. After developing a hypothesis or investigating a disease, groups gave oral presentations and submitted a written report explaining their collaborations and findings. By the end of the semester, each team of experts had collaborated with every other team, thereby investigating relationships between their system and all other physiological systems.
Factors influencing variation in the diameter of anterior and posterior intercostal arteries
Authors: Lane Richard Durelle , Baptista Carlos , Hankin Mark , Bennett‐Clarke Carol
Published: June 2021

The anterior and posterior intercostal arteries are the principal blood supply to the intercostal spaces and ribs. Each intercostal space is supplied by an anastomosing anterior and posterior intercostal artery, as well as their collateral branches. These anastomoses can provide important alternate routes of blood flow for individuals with clinical conditions such as a coarctation of the aorta, or when an internal thoracic artery is harvested for a coronary bypass graft. The clinical significance of these vessels raises several questions regarding their relationships, such as: Is there a correlation between the relative diameters of the ipsilateral paired intercostal arteries under normal conditions?; Is there a pattern to the changes in vessel diameter in different intercostal spaces, particularly comparing superior with inferior spaces?; Do the left posterior intercostal arteries increase in diameter when the left internal thoracic artery has been harvested for a bypass graft? In this study, cadaveric paired samples of the 2nd through 6th anterior and posterior intercostal arteries were collected and their diameters measured. Initial data analysis suggests that both anterior and posterior intercostal arteries increase in diameter as one progresses from the 2nd to the 6th intercostal spaces. Analysis of these vessels will be correlated with specific clinical conditions noted in each cadaver.
SEGMENTATION OF LIVER CAUDATE LOBE. STUDY ON CORROSION CASTS
Authors: Matusz Petru L. , Hordovan Eniko‐Christine
Published: June 2021

Terminologia Anatomica (1998) considers liver's caudate lobe (CL) as an independent volume of parenchyma, described as the dorsal portion of the liver – Segment I. In the anatomic literature there are a number of descriptions of CL segmentation. Rex (1888), Hjorstjo (1948), Couinaud (1954 and 1957), Reiferscheid (1957), Diaconescu (1963), Platzer (1988), Terminologia Anatomica (1988), Lanz and Wachsmuth (1993) and Terminologia Anatomica (1998) describe CL as a unique segment. Ciobanu (1958) describes 2 segments at the level of CL. Healey et al (1953), Couinaud (1998), Kogure et al (2000) describe 3 segments of CL parenchyma. We analyzed the segmental distribution of the portal branches of CL and the disposition of the fissural planes of venous drainage for the CL on 150 hepatic corrosion casts. They were made by injecting with plastic of the vasculo‐ductal systems, followed by parenchyma corrosion. The distribution of the portal branches of the CL reveals the presence in the parenchyma of the CL of two segments (papillary process and paracaval portion) in 82% cases, and of three segments (papillary process, paracaval portion and caudate process) in 18% cases. These aspects must be considered when planning hepatic surgery (Supported by CNMP 41092/2007).
Distribution of mucusal surface area in nasal and paranasal spaces of a monkey and mouse lemur
Authors: Smith Timothy D , Eiting Thomas P , Rossie James B
Published: June 2021

Internal nasal anatomy differs markedly between the two major groups of primates. Strepsirrhines (e.g., lemurs) have more numerous turbinals and recesses compared to haplorhines (e.g., monkeys). However, no detailed quantitative comparisons have been made. Using adult mouse lemurs (Microcebus sp) and tamarins (Saguinus sp), we measured nasal and paranasal surface area (SA) in two serially sectioned heads and digitally reconstructed the airways using two dry crania. Olfactory SA was also quantified. Measurements reveal that mucosal SA on turbinals is 17% greater in Microcebus than in Saguinus. The ethmoturbinal complex of Microcebus accounts for half of the total olfactory mucosa in the nasal airway. In contrast, most (61%) olfactory mucosa in Saguinus is distributed on other surfaces (e.g., nasal septum). Paranasal spaces account for 17% of total internal nasal SA in Microcebus and 26% in Saguinus, perhaps revealing different degrees of secondary pneumatization. Taken together, these results suggest different patterns of mucosal distribution between haplorhines and strepsirrhines. The distribution of mucosa between these two groups may reflect differences in airflow through the nose, and particularly, into and out of the olfactory region. Under the constraint of a reduced ethmoid region, haplorhines may have redistributed, rather than reduced, olfactory mucosa.

This study was funded by the National Science Foundation, # BCS‐0820751.