bluuebunny/crossref_metadata_2025 huggingface dataset:
Does hypertension affect morphometric parameters of phrenic nerves?
Authors: Rodrigues Anaceres Ribeiro , da Silva Ferreira Renata , Salgado Helio Cesar , Fazan Valéria Paula Sassoli
Published: June 2021

We previously reported [1] important differences in phrenic nerve morphology between normotensive Wistar‐Kyoto and spontaneoulsy hypertensive rats (SHR) but the question whether the differences are due to hypertension or intrinsic to the rat strains still remains. We aimed to investigate the phrenic nerve morphology in young normotensive SHR and adult SHR with well established hypertension. Female SHR aged 5 (N=6) or 20 (N=6) weeks were anesthetized for arterial pressure (AP) recording. Proximal and distal segments of the right and left phrenic nerves were removed and prepared for epoxy resin embedding and light microscopy morphometry. Adult rats showed significantly higher body weight and AP, compared to young animals. Phrenic nerves showed morphologic and morphometric lateral and longitudinal symmetry for both groups but were significantly larger on older animals. Myelinated fiber area and diameter were also larger on older animals but their number was not different between groups. Myelinated axons were larger on older animals but the G ratio was not different between groups. As expected, aging affected phrenic nerve fascicle and myelinated fibers size with no alteration in fiber number. Also, hypertension development did not affect morphological parameters considered important for myelinated fiber function.

Grant Funding Source : FAPESP, CNPq, CAPES and FAEPA
Neuronal counting in the spinal cord dorsal horn of male adult Wistar rats after noxious stimuli during neonatal period
Authors: Machado Nathalia Leilane Berto , Sanada Luciana Sayuri , do Carmo Elisabete Cássia , de Oliveira Alexandre Leite Rodrigues , Fazan Valéria Paula Sassoli
Published: June 2021

The aim of the present study was to analyze the effects of noxious stimuli during neonatal period in the neuronal distribution in the spinal cord dorsal horn in adult rats. Six male Wistar rats with 180 days of life were separated in 2 groups: pain group (n=3) and control group (n=3). The pain group was stimulated with a needle on the right paw and the control group was stimulated with a cotton swab on the right paw, being both groups stimulated twice a day for 15 days. Were evaluate the hyperalgesia of both groups with Von Frey Monofilament and a specific calibrated forceps; the analysis of the neuronal distribution was performed by counting Nissl stained neurons with a specific software. Statistic tests were applied, and p value was considered significant when p was less than 0,05. We observed that the pain group presented a hyperalgesia in the right hindpaw when compared to the control group, but no differences were found in the neuronal counting in the spinal cord between groups. Our results suggest that noxious stimuli during neonatal period are capable of changing the sensorimotor system in the long term, but this change does not involve changes in the neuronal distribution in the dorsal horn.

Grant Funding Source : FAPESP, CNPq and FAEPA
The effect of quercitin on oxidative stress‐induced bone‐like cells (UMR 106‐01 BSP)
Authors: McCowan Brandon , Bria Mariana , Joseph Eugene E.
Published: June 2021

Antioxidants are reported to play a role in bone metabolism, prevention of bone loss, and the reduction of oxidative stress. A reasonable amount of research has been conducted linking quercetin, a flavonoid antioxidant, with treatment of cancer, disorders of skin and connective tissue. However, not much research has been done connecting quercetin with bone health. The purpose of this study was to determine whether quercetin exhibits a protective effect on UMR bone‐like osteosarcoma cells (UMR 106‐01 BSP) by decreasing the effects of oxidative stress in these cells. To test this, UMR cell cultures were co‐treated with differing concentrations of quercetin and 85μM of an oxidant, buthionine sulfoximine (BSO). Caspase‐3 and NF‐ κB (p65 subunit) analyses were performed to measure cell viability and nuclear activity, respectively, using a flow cytometer. F test ANOVA was used to compare variances and all values in the results were expressed as means ±SD. In all cases, p values of <0.05 were considered statistically significant. The results showed that quercetin effectively protected bone‐like cells against the destructive effects of oxidative stress at an optimal concentration very close to 50μM. As quercetin concentrations increased in co‐treated cell cultures, caspase‐3 levels decreased and NF‐κB activity increased. This suggests that quercetin protects bone cells from undergoing apoptosis due to oxidative stress in a dose‐dependent manner by inhibiting caspase‐3. In addition, quercetin may influence the NF‐κB pathway to alter DNA transcription in the nucleus. These findings may help in the elucidation of the potential benefits of quercetin on bone health. (Supported by the Dean's Office of the College of Arts and Sciences of La Sierra University, Riverside, CA).
Zygapophyseal joint space gapping following spinal manipulation in low back pain patients
Authors: Cramer Gregory D , Cambron Jerrilyn A , Cantu Joe A , Dexheimer Jennifer , Pocius Judith D , Gregerson Douglas , Fergus Michael , McKinnis Ray A
Published: June 2021

This study quantified lumbar zygapophyseal (Z) joint space separation (therapeutic gapping) in acute low back pain subjects after spinal manipulative therapy (SMT) or side‐posture positioning (SPP).

Methods

This IRB‐approved project enrolled 112 acute low back pain subjects. After 2 weeks of treatment all subjects received an MRI scan (supine position) of the L4/L5 and L5/S1 Z joints. Subjects were then randomized into SPP (non‐manipulation) Groups 1 or 4 (n=28, 26) or SMT (manipulation) Groups 2 or 3 (n=26, 26). SMT and SPP were performed with the most painful side of the lumbar region up. After the SMT or SPP procedure/intervention, a second MRI scan was taken with subjects either supine (Groups 2 and 4) or maintaining the SPP (Groups 1 and 3). Observers blinded to group and MRI scan (first or second) measured the central anterior‐posterior (A‐P) joint space on the scans. Difference between A‐P measurements taken post‐ and pre‐intervention was the Z joint “gapping difference.” Gapping differences were compared among and between groups (ANOVA).

Results

Final data analysis was performed on 106 subjects (51 male and 55 female). Differences were significant (p=0.0005; Group 1=0.6 +0.9mm, Group 2=0.2 +0.4, Group 3=0.9 +0.7, Group 4=0.4 +0.3).

Conclusions

SMT (Group 3) resulted in the greatest amount of therapeutic gapping in acute low back pain subjects. Funding: NIH/NCCAM Grant # 2R01AT000123.
Kinetically differentiated post‐translational modifications in a transcription factor, Bcl11b
Authors: Filtz Theresa M , Vogel Walter K , Zhang Ling-juan , Liu Xiao , Arbogast Brian , Zweber Adam , Maier Claudia S , Leid Mark
Published: June 2021

Bcl11b is a highly modified transcription factor that is required for T cell development. Stimulation of primary thymocytes through the MAP kinase pathways leads to a series of post‐translational modifications of Bcl11b that ultimately alter its function from a repressor to a derepressor of a model target gene. These modifications include hyperphosphorylation and desumoylation at 5 min, significant dephosphorylation at 30 min and then subsequent resumoylation at 60 min, followed ultimately by ubiquitination‐mediated degradation at 4 to 8 hours. We sought to define the sites of Bcl11b phosphorylation and sumoylation to better understand the role of these modifications in integrating the signals from extracellular stimulation into alterations of the gene network in developing thymocytes. We identified 23 sites of phosphorylation and one site of sumoylation by mass spectrometry analysis. We identified a second sumoylation site by site‐directed mutagenesis. For the phosphorylation sites, we have begun to define the kinetics of each site following thymocyte stimulation. Thus far, four distinct kinetic patterns are observed among the 23 sites. Further, we have found differential association of the activated ERK kinase and PP6 phosphatase subunits in complex with Bcl11b. (This work was supported by the NIH, grant R15 GM096243, and the OSU College of Pharmacy.)
Effects of alkyl chain length of Zn N‐alkylpyridylporphyrins on photo‐mediated protein crosslinking
Authors: Craik James , Ghazal Monthir , Batinic-Haberle Ines , Benov Ludmil
Published: June 2021

Photodynamic therapy provides a minimally invasive approach for cancer treatment and Zn(II) N‐methylpyridylporphyrins can act as potent photosensitizers (PSs). Lipophilicity is a major factor determining transmembrane permeation of porphyrins and may influence their association with proteins. This study used SDS‐PAGE to investigate damage to purified lactate dehydrogenase and isolated human erythrocyte membranes by amphiphilic butyl and hexyl Zn(II) N‐alkylpyridylporphyrins under PS illumination conditions that kill cultured cancer cells. Localization of PSs and role of singlet oxygen were examined by using scavengers (L‐histidine and sodium azide), D2O, and hypoxia. Increased alkyl chain length gave increased photo‐crosslinking of membrane proteins. Significance of binding of Zn(II) N‐alkylpyridylporphyrins to proteins was examined by addition of photo‐inactive Mn N‐alkylpyridylporphyrin analogues. Limited PS‐dependent protein modification was found under hypoxic conditions and PS treatment of L‐histidine generated different products under aerobic and anaerobic conditions. SDS‐PAGE profiles suggested that rapid protein modification occurs at the cytosolic face of the erythrocyte membrane and that protein crosslinking is not favored between domains within membrane core regions. Hematoporphyrin D was much less potent than the Zn‐PSs in protein photo‐crosslinking. Financial support from Grant YM 02/08 from Kuwait University
Family 18 Chitinase from Bacillus pumilus SG2: Role of Chitin‐Binding Domain (ChBD) in the Enzymatic Activity
Authors: Ghasemi Seyedhadi , Amiriahmadvandi Maryam , Karimi Amirhossein
Published: June 2021

The Bacillus pumilus SG2 chiS and chiL genes, encode chitinases S (ChiS) and L (ChiL). Chitinase S consists of an open reading frame of 1,788 nucleotides and encodes 596 amino acids with a deduced molecular weight of approx. 66 kDa. The ChiS is an enzyme composed of a glycosyl hydrolases family 18 domain responsible for chitinase activity, one fibronectin type III domain, and a C‐terminal chitin‐binding domain (ChtBD). Chitinase L contains a representative of GH‐18 domain and ChiC (Chitinase C) domain. To delineate the properties of ChtBD domain, truncated form of chitinase S (ChiSΔC), lacking the chitin‐binding domain, was generated and expressed in Escherichia coli strain M15 cells using pQE‐30 vector. The hydrolytic activities toward insoluble and soluble substrates were significantly reduced by the truncation of ChtBD and ChiL. In addition, antifungal activity determined from the digestion rate of hyphae of fungal phytopathogens was reduced by the ChtBD truncation and ChiL. The results indicate that chitin‐binding domain plays an important role in the enzymatic activity of ChiS.
Hydrogen sulfide (H2S) regulates hypoxic signaling in T cells
Authors: Miller Thomas W , Song Timothy , Amarnath Shoba , Fowler Daniel H , Roberts David D
Published: June 2021

Hydrogen sulfide (H2S) is emerging as an important member of the gasotransmitter family that includes nitric oxide and carbon monoxide. We have recently reported that H2S is an important immunomodulatory molecule that potentiates the activation of human and murine T cells. As H2S signaling in T cells plays a vital functional role, it is critical to identify the relevant signaling target(s) and define the role of H2S in T cell differentiation and effector function. We have found that nanomolar levels of H2S dose‐dependently enhance HIF1α protein levels in activated T cells by 65% under hypoxic conditions. Under these conditions, H2S also enhanced VEGF gene expression. Consistent with recent reports of HIF1α‐dependent T cell differentiation, we observed an increase in Th17 cells and a decrease in Treg cells when splenic T cells were differentiated in the presence of 1000 nM H2S. We are currently characterizing the regulation of hypoxic T cell metabolism by H2S and the proximal target of H2S in this hypoxic response pathway. This work has exciting implications for understanding the progression of inflammatory bowel disease and for development of antitumor immune therapies.
Catalase overexpression and mitochondrial function in insulin resistant muscle cells
Authors: Silveira Leonardo Reis , Sampaio Igor H , Bomfim Lucas , Queiroz André , e Sousa Thais Amaral , Gonzaga Bruno , Zoppi Claudio , Carneiro Everardo Magalhães , Curi Rui
Published: June 2021

We examined whether the reduction of mitochondrial capacity and insulin response imposed by the high availability of fatty acids are associated with an increase of intracellular hydrogen peroxide (H2O2). We investigated whether the oxidative stress induced by excess fatty acids can be reversed with improved antioxidant capacity through overexpression of catalase. The catalase gene was successfully transfected in skeletal muscle cells through pcDNA3 plamid as indicated by catalase mRNA and content (p<0.05). The H2O2 production was increased after palmitic acid treatment. This effect markedly reduced the citrate sintase activity as well as the mRNA levels of PGC1α (p<0.05). This effect was accompanied by a significantly reduction of p‐AKt which further was demonstrated to down regulate p‐CREB and PPAR‐β. In addition, oxygen consumption and uptake glucose were markedly reduced in the presence of palmitic acid (p<0.05). However, the catalase transfection was observed to prevent reduction of p‐AKt level which further was demonstrated to up regulate p‐CREB and PPAR‐β (p<0.05). Our results showed that fatty acid reduced the p‐AKt level. This effect was associated with a low p‐CREB level and consequently PGC1‐α transcription. The reduced mitochondrial oxygen consumption favored ROS production and insulin resistance. The catalase‐ induced effect on p‐AKt increased the oxygen consumption and glucose and fatty acid metabolism suggesting that mitochondrial biogenesis might be regulated by phosphorylated level of AKt.
Identification of the essential cell wall stress sensors in a Saccharomyces cerevisiae budding yeast lacking a myosin heavy chain gene (myo1Δ)
Authors: Santiago-Cartagena Ednalise , González-Crespo Sahily , Rodríguez-Medina José R.
Published: June 2021

The PKC1‐MAPK pathway in Saccharomyces cerevisiae is activated by stress sensor proteins Wsc1‐3, Mid2, and Mtl1, the plasma membrane proteins responsible for sensing different types of cell wall and physiological stress for maintenance of cell wall integrity. A deletion mutant of MYO1 (myo1Δ) encoding the myosin II heavy chain, has a primary defect in cytokinesis and structural deficiencies in the cell wall that activate the PKC1‐MAPK pathway. We proposed to investigate which of these cell wall stress sensors are involved in PKC1‐MAPK pathway activation in myo1Δ strains. Double mutants containing myo1Δ and a deletion of each stress sensor gene were constructed and tested according to cell morphology and viability phenotypes, as well as the level of activation of the PKC1‐MAPK pathway. The results showed that each of these double mutants displayed a cell morphology similar to myo1Δ. The wsc1Δmyo1Δ and wsc3Δmyo1Δ strains presented a severe synthetic growth defect. PKC1‐MAPK activity levels indicated that Wsc1p, Wsc3p, and Mid2p were the primary sensors involved in activation of this pathway. In conclusion, cell wall stress sensor proteins of the PKC1‐MAPK pathway can detect stress caused by the myo1Δ defect in cytokinesis at the cell wall. Supported by grants from NIAID/NIGMS (SC1AI081658), NCRR‐RCMI (G12RR003051) and MBRS‐RISE (R25GM061838).
Genetic depletion of the phosphatase PHLPP1 upregulates receptor tyrosine kinase signaling
Authors: Reyes Gloria Ximena , Newton Alexandra
Published: June 2021

The PH‐domain leucine rich repeat protein phosphatase PHLPP1 is a Ser/Thr phosphatase that negatively regulates Akt signaling. This pathway controls many cellular processes; loss of PHLPP leads to hyperactivation of Akt and an imbalance in cell homeostasis, triggering many diseases. Using cells derived from a phlpp1−/− mouse, we show that PHLPP1 suppresses the amplitude of the ERK pathway through an Akt‐independent mechanism. phlpp1−/− cells have increased receptor tyrosine kinase levels leading to augmented activation of downstream signaling. In these cells, the effects on ERK are due to an increased rate of receptor biosynthesis, not defective receptor degradation. Neither pharmacological inhibition of the PI3K‐Akt pathway, nor the MEK‐Erk pathway affects receptor levels, indicating that the PHLPP1 target is not Akt nor ERK. Furthermore, the catalytic and leucine rich repeat regions are critical to regulate receptor levels in normal cells. To find the mechanism controlling receptor levels, we performed pharmacological and PCR arrays to dissect the components responsible for this phenotype in phlpp1−/− cells. Our data indicate that general transcriptional machinery rather than only one transcriptional factor are involved. These data unveil an additional function of PHLPP1 in suppressing receptor tyrosine kinase signaling in cells by a transcriptional mechanism.

This work was funded by NIH GM067946.
Targeting the Metastasis Suppressor NDRG1: A New Strategy for the Treatment of Pancreatic Cancer
Authors: Richardson Des R , Chikhani Sherin , Lui Goldie , Sivagurunathan Sutharshani , Kovacevic Zaklina
Published: June 2021

Aims

The metastasis suppressor, N‐myc downstream regulated gene‐1 (NDRG1), is negatively correlated with tumor progression in pancreatic cancer. Moreover, NDRG1 is an iron‐regulated gene that is markedly up‐regulated by cellular iron‐depletion using novel anti‐tumor agents such as the chelator, Dp44mT, in pancreatic cancer cells. However, the exact molecular function(s) of NDRG1 remain to be established.

Results

Using gene‐array analysis together with NDRG1 over‐expression and silencing, we identified molecular targets of NDRG1 in 3 pancreatic cancer cell lines. For the first time, we demonstrate that NDRG1 up‐regulates neural precursor cell expressed developmentally down‐regulated 4‐like (NEDD4L) and GLI‐similar 3 (GLIS3). Further studies examining the down‐stream effects of NEDD4L led to the discovery that NDRG1 affects the transforming growth factor β (TGF‐β) pathway, leading to up‐regulation of two key tumor suppressor proteins, namely phosphatase and tensin homologue deleted on chromosome 10 (PTEN) and mothers against decapentaplegic homolog 4 (SMAD4) in multiple cell lines. Moreover, NDRG1 inhibited the phosphatidylinositol 3‐kinase (PI3K) and Ras oncogenic pathways.

Conclusion

The identified target genes of NDRG1 and their effect on the TGF‐â signaling pathway reveal its molecular function in pancreatic cancer and a novel therapeutic avenue.
Efficient down‐regulation of glia maturation factor expression in brain and spinal cord
Authors: Zaheer Asgar , Wu Yanghong , Thangavel Ramasamy , Sahu Shailendra K , Zaheer Smita
Published: June 2021

Long‐lasting siRNA‐based down‐regulation of gene of interest can be achieved by lentiviral‐based expression vectors driving the production of short hairpin RNA (shRNA). We investigated an attractive therapeutic approach to target the expression of proinflammatory GMF by using lentiviral vector encoding GMF‐specific shRNA to reduce GMF levels in the spinal cord and brain of mice. To determine the effect of GMF‐shRNA on GMF protein levels, we performed quantitative ELISA analysis in brain and in thoracic, cervical and lumbar regions of spinal cord from mice followed by GMF‐shRNA (G‐shRNA) or control shRNA C‐shRNA) treatments. Our results show a marked reduction of GMF protein levels in brain and spinal cord of mice treated with GMF‐shRNA compared to control shRNA treatment. Thus, the down‐regulation of GMF by GMF‐shRNA was efficient and wide spread in CNS as evident by the significantly reduced levels of GMF protein in the brain and spinal cord of mice
Protein kinase C α phosphorylates argininosuccinate synthase at serine 328 in endothelial cells to affect nitric oxide production
Authors: Haines Ricci , Corbin Karen , Pendleton Laura , Eichler Duane
Published: June 2021

Nitric oxide (NO) signaling is required for vascular function and its homeostasis is altered in atherosclerosis and/or vascular complications of diabetes. The recycling of citrulline to arginine creates a distinct pool of arginine critical for NO production and is known as the citrulline‐NO cycle. Argininosuccinate synthase (AS) is the rate‐limiting enzyme for NO production, and therefore its regulation affects NO synthesis. Our laboratory has recently identified a biologically relevant AS phosphorylation site at serine 328 that responds to physiological cues in cultured bovine aortic endothelial cells (BAECs). In this report, we describe an investigation into the relevant pathways that regulate AS function through the phosphorylation of serine 328. We show by kinase inhibition, kinase knockdown, and in vitro kinase activity assays that protein kinase C α (PKCα) phosphorylates AS at serine 328 as the result of calcium dependent stimulation of NO in BAECs. In addition, we show signaling pathways that do not involve PKCα, such as insulin or VEGF signaling, decrease phosphorylation of serine 328. In conclusion, these data suggest that PKCα mediated phosphorylation of serine 328 occurs under conditions that promote NO production in response to distinct physiological cues that are dependent on calcium signaling in endothelial cells.
Resolving the structure of Bcy1, the Regulatory Subunit of Yeast Protein Kinase A
Authors: Núñez Lucas Matías Fernández , Bardeci Nicolás González , Rossi Silvia , Blumenthal Donald , Moreno Silvia
Published: June 2021

The major receptor for cAMP signaling in eukaryotes is the regulatory subunit (R) of protein kinase A. Saccharomyces cerevisiae has one R (Bcy1). R subunits are homodimers comprising an N‐terminal dimerization/docking domain (D/D), an inhibitory hinge region, and two tandem cAMP binding domains (CNB) at the C‐terminus. The crystal structure of Bcy1(168–416) has been solved. The relative orientation of the two CNB domains is different from RIα and RIIβ. We have analyzed the solution structure of Bcy1 whole molecule and of a monomer (Δ140)Bcy1 by small‐angle x‐ray scattering (SAXS). The Rg (gyration radius) and dmax (derived from the P(r) function) of the molecules were determined. Both structures were modeled ab initio using the DAMMIN program. The resulting structure of (Δ 140)Bcy1 compares very favorably with the crystal structure, thus confirming the particular relative orientation of the CNB domains The structure of the whole homodimer is predicted to be different from RI and RII. The N terminus of Bcy1 was modeled by homology using Swiss Model and further minimization with the GROMACS package. It shares with its counterparts the residues important for dimerization; however the exposed surface, that should interact with anchoring proteins (not described yet in yeast), has characteristics of its own.
Control of cortical actin assembly and cadherin‐catenin localization by GPCRs and RhoGTPases
Authors: Jimenez-Dalmaroni Maximiliano Javier , Heasman Janet , Wylie Christopher
Published: June 2021

Many common and permanently disabling human birth defects, such as spina bifida, are caused by the loss of cell shape changes and tissue movements in the early embryo. Therefore, it is important to understand how these cell shape changes are regulated during embryonic development. It has been previously shown that the actin cytoskeleton is required for normal tissue movements at these stages. Furthermore, the expression and localization of cadherin‐catenin complex are relevant for cell adhesion and F‐actin assembly and their lost can increase risk of metastasis in carcinomas and alter normal cell movements during embryonic development.

Using the Xenopus laevis embryo and human embryonic stem cells as models, we are currently investigating the mechanisms by which cortical actin assembly is regulated during development and the signaling pathways relevant for the cadherin‐catenin localization. We have identified two G‐protein couple receptors, a novel GPCR receptor termed xFlop and xLPA, and the small GTPase Rac1, as key molecules involved in this process. We hope this research could lead us to not only better understanding of cortical actin assembly and remodeling in early stages of Xenopus laevis embryos and humans, but also to develop novel treatments for spina bifida and human cancer metastasis.
The N‐terminal actin binding domain (ABD1) of dystrophin is in a closed conformation in solution and undergoes a conformational transition upon binding to F‐actin
Authors: Mallela Krishna , Singh Surinder
Published: June 2021

Deficiency of the vital muscle protein dystrophin triggers Duchenne/Becker muscular dystrophy. Dystrophin stabilizes muscle cells against forces associated with muscle contraction and stretch. It interacts with F‐actin using its N‐terminal actin‐binding domain (N‐ABD or ABD1), which is comprised of tandem calponin‐homology (CH) domains. A previously determined X‐ray crystal structure indicated that the N‐ABD is a domain‐swapped dimer, with each monomer adopting an ‘open’ conformation in which the two CH domains do not interact. This structure is controversial as it contradicts known structures of similar domains in other muscle proteins. Here we investigated the conformation of dystrophin N‐ABD in solution using a simple and elegant technique of pyrene excimer fluorescence. Our results clearly indicate that dystrophin N‐ABD is a monomer in solution and is in a ‘closed’ conformation in which the two CH domains appear to interact. Upon binding to F‐actin, this domain transitions into a more compact closed conformation. This study demonstrates for the first time the conformational transition of a dystrophin domain during function, and suggests that such structural dynamics play important role in controlling dystrophin function.
Clinically Oriented Anatomy and Physiology for a Master Physician Assistant Program
Authors: Escobar-Poni Bertha Corina
Published: June 2021

All that hard work behind a syllabus is very rarely perfection; in the end, is just ‘clay’ to be reshaped by the needs and expectations of both students and teachers. The previous statement summarizes my recent experience teaching a clinically oriented anatomy and physiology course to students in our Master Degree program for Physician Assistant. My first quarter was a disappointment, but the 2nd quarter was a success (based on student's evaluations). Although I did little changes in the online physiology modules, the greatest change was in the clinical anatomy section. The hallmark of the success revolved around two aspects: teamwork, and empowering through active learning. The implementation of three learning methods made a big difference: rotating teaching stations, training experts, and peer teaching. A student from each team was trained to become an expert in specific anatomic territory including clinical applications, thus be a resource in their teams. Each student took seriously the responsibility for their peers learning. Students left every lab with a sense of accomplishment, confident about how much they learned and making plans to meet after hours. Last year's learned lessons of dos and don'ts were pivotal to a great start with this new class. I am looking forward to our mutual learning and sharing.

Support: Henry Gray/Elsevier Distinguished Educator Award Plenary Symposium
Attenuated L1 expression by glioma cells results in decreased cell migration on monolayer co‐cultures
Authors: Bhatti Hamza , Galileo Deni
Published: June 2021

L1 is a neural recognition molecule primarily responsible for the invasive quality of glioblastoma multiforme (GBM) cells. It acts by signaling through integrin receptors, and more recently discovered, the fibroblast growth receptor (FGFR). In addition, L1 is abnormally expressed, proteolyzed by ADAM10, and released (shed) by high‐grade glioma cell lines. Using short‐hairpin RNA (shRNA) and recombinant DNA techniques, we used lentiviral vector to shut down L1 expression in a L1 expressing glioma cell line (T98G). The effect of attenuated L1 expression by glioma cells on cell motility was measured on a substratum of axons and glial cell monolayer isolated from embryonic chick brain tissue. The results show that glioma cells that secrete L1 (T98G‐pLKO.1) display greater motility on monolayer co‐cultures than glioma cells that do not secrete L1 (T98G‐shL1). Research support was provided by Howard Hughes Medical Insititute.
Molecular analysis and modeling of inactivation of CYP2D6 by four mechanism‐based inhibitors
Authors: Livezey Mara R. , Nagy Leslie D. , Diffenderfer Laura E. , Arthur Evan J. , Hsi David J. , Furge Laura Lowe
Published: June 2021

There are four known and confirmed mechanism‐based inactivators of human cytochrome P450 2D6: SCH 66712, EMTPP, paroxetine, and 3,4‐methylenedioxymethamphetamine (MDMA). SCH 66712 and EMTPP contain piperazine groups and substituted imidazole rings; previous studies with each have indicated that inactivation occurs by reaction with the substituents of the imidazole ring. Paroxetine and MDMA contain methylenedioxyphenyls; the moiety responsible for inactivation is unknown. The current study shows that each inactivator displays Type I binding with Ks values that vary by 2‐orders of magnitude. Comparison of Ki and kinact and partition ratio values shows Schering 66712 is the most potent inactivator. Molecular modeling experiments using AutoDock identify Phe120 as a key interaction for all four inactivators with face‐to‐face and edge‐to‐edge pi interactions apparent. Also, modeling suggests Thr309 could be a potential site for inactivation. Ligand conformations were scored according to their binding energies as calculated by AutoDock and correlation was observed between molecular models and Ks values. (Support: NIH 1R15‐GM086767‐01 & ‐01S1; HHMI [52006304] to Kalamazoo College).
Major Facilitator Superfamily Domain‐containing protein 2a is a novel regulator of hepatic lipid metabolism
Authors: Berger Justin H , Silver David L
Published: June 2021

Understanding how the body switches between storing and burning energy is a fundamental gap in our ability to better treat metabolic disease. The Major Facilitator Superfamily Domain‐containing protein 2a (MFSD2A) is a newly identified plasma membrane transporter that is nutritionally regulated in liver and brown adipose tissue. MFSD2A protein expression is not detectable in the livers of fed mice, but is highly induced during a fast in a PPARalpha‐ and glucagon receptor‐dependant manner. Additionally, MFSD2A turns over rapidly within 60 minutes, and disappears from the cell surface with similar kinetics. In order to investigate whether MFSD2A plays a role in lipid metabolism, we generated a conventional Mfsd2a knockout mouse model. In the absence of MFSD2A, normal triglyceride accumulation in the liver during a fast was not observed. Additionally, knockout mice exhibited increased energy expenditure and protection from diet‐induced weight gain, without changes to glucose homeostasis. Further data on the mechanism of MFSD2A will be presented. Taken together, our studies indicate that MFSD2A plays an important role in regulation of energy expenditure and liver triglyceride levels.
Biochemical Characterization Of PMM2‐depleted Zebrafish Suggests An Unexpected Mechanism For Glycosylation Deficiency In CDG‐Ia
Authors: Lehrman Mark A. , Gao Ningguo , Cline Abigail , Flanagan-Steet Heather , Freeze Hudson H. , Sadler Kirsten C. , Steet Richard
Published: June 2021

Congenital Disorder of Glycosylation (CDG) Type Ia is caused by mutation of the PMM2 gene encoding phosphomannomutase, which converts mannose‐6‐P (M6P) to mannose‐1‐P (M1P). Patients have abnormalities associated with impaired protein N‐glycosylation attributed to diminished M1P, a precursor of GDP‐mannose and the lipid‐linked oligosaccharide (LLO) needed for N‐glycosylation. We used pmm2‐directed morpholinos to develop a CDG‐Ia model using zebrafish, with reduced phosphomannomutase activity and developmental abnormalities consistent with CDG‐Ia. By Fluorophore‐Assisted Carbohydrate Electrophoresis, we observed losses of LLOs (58% of control) and total N‐glycans (67% of control) as expected for CDG‐Ia. However, the pmm2 depletion was insufficient to reduce M1P and GDP‐mannose concentrations. This suggested the CDG‐Ia phenotype resulted from another aspect of Pmm2 depletion. Aside from M1P loss, Pmm2 deficiency might also increase the enzyme's substrate, M6P. We reported recently that M6P signals in a novel pathway causing destruction of LLO molecules during viral infection. In pmm2 morphants, M6P was indeed elevated (187% of control), and surprisingly free glycans (products of LLO destruction) were increased (195% of control). Phenotypes were rescued by co‐injecting pmm2 mRNA. Thus, M6P elevation may cause LLO depletion in our zebrafish CDG‐Ia model. Supported by NIH‐ARRA RC1‐HD064159.
Apoptotic mechanism of Bisphenol A in human neuroblastoma
Authors: Park Bongkyun , Rhee Dong-kwon , Pyo Suhkneung
Published: June 2021

Bisphenol A (BPA) which is known to be a xenoestrogen and induces apoptosis in various cell lines is an endocrine disruptor of nervous system. Peptidylarginine deiminase IV (PADI4) is well known for immune cell differentiation and apoptosis. However, the precise mechanisms of BPA‐induced apoptosis remain unclear. In the study, we investigated the role of PADI4 in BPA‐induced neuronal cell death. Western bloting showed that BPA induced activation of caspase‐3 and apoptosis‐related genes (p53, Bax, and Bid) in a concentration dependent manner. Our data also showed that BPA increased the generation of ROS and phosphorylation of JNK and p38. MTT assay revealed that pretreatment with specific inhibitors for ROS, JNK and p38 reversed BPA‐induced cell death. Western bloting also showed that BPA increased protein levels of PADI4, whereas pretreatment with inhibitors of ROS, JNK and p38 decreased PADI4 and Bid expression induced by BPA. In addition, immunoprecipitation analysis demonstrated that BPA treatment resulted in increase of interaction between PADI4 and p53. Inhibition of PADI4 activity by siRNA also suppressed BPA‐induced p53 expression and apoptosis. Taken together, these results provide more insight into the mechanism of Bisphenol A‐induced neurodegeneration.
Factors associated with weight change up to 12 months postpartum in HIV‐infected women: findings from the HIV prevention of mother‐to‐child transmission (PMTCT) Kesho Bora trial
Authors: Cames Cecile , Cournil Amandine , Bork Kirsten
Published: June 2021

Data on postpartum weight change (PWC) in HIV‐infected women are sparse and inconsistent. This study analyzed, according to antiretroviral prophylaxis (ARV), clinical and infant feeding (IF) characteristics, PWC from 1 to 12 months in HIV‐infected women participating in a PMTCT trial in 5 study sub‐Saharan African sites.

Women with 200–500 CD4+ cells/mm3 were randomized to receive from 28–36 weeks gestation either a triple ARV (3 ARV), continued up to 6 months postpartum (PP) or a standard ARV (STANDARD) until 1 week PP.

Among 792 women, 78% breastfed for a median duration of 5.1 months (IQR: 2.8–6.0). At delivery, viral load (VL) was undetectable in 64% of 3 ARV women vs. 30% in STANDARD group (P<0.0001) and median CD4 count was 463 and 414 cells/mm3, respectively (P<0.0001). At 1 month PP, less than 3% were underweight while 49% were overweight or obese. Mean PWC at 12 months was +1.4kg in 3 ARV group and −0.7kg in STANDARD group (P<0.0001). In mixed linear model analyses, 3 ARV, baseline overweight or obesity and C‐section were independently associated with PWC (+65, +54 and +56 g/month, P<0.0001, <0.001 and =0.02, respectively). No association was found between either IF or CD4 count or VL and PWC.

3 ART increased weight gain over the first year PP and IF had no impact on PWC among these HIV‐infected women with intermediate advanced disease.

Funding: ANRS, DFID, ECDTP, Trasher Research Fund, GSK Fund, CDC, NIH, WHO
Relation between Dairy Calcium Intake, Bone Mineral Density and Lipids in Obese Post‐Menopausal Women
Authors: Mamed Narmina , Beeson Lawrence , Fakhrawi Dina , Darnel Alan , Cordero-MacIntyre Zaida
Published: June 2021

Objective

To assess the relationship between lipids and BMD in obese postmenopausal women placed on a dairy calcium weight‐reduction diet.

Methods

Analysis of data performed on 56 obese postmenopausal women (mean age: 55.61±8.19; mean BMI: 32.95±6.12 kg/m2; mean weight: 86.88±17.25; and mean BMD level: 1.05±0.17 gms/cm2) randomly distributed into a Low Calcium (800 mg/d) or High Calcium (1400 mg/d) diet to evaluate differences in BMD, body mass index (BMI) and lipid profiles (total cholesterol (TC), low‐density lipoproteins (LDL), high‐density lipoproteins (HDL), and triglycerides(TG)).

Results

Significant reduction for combined treatments in BMI : −0.72±1.02 kg/m2, 95% CI: (−0.99, −0.44) (p <0.001); LDL : −18.14±33.55 mg/dL, 95% CI (−27.12, −9.15) (p <0.001); and TC : −81.82±139.59 mg/dL, 95% CI (−137.04, −26.59) (p=0.005). Significant increase in HDL : 3.79±11.51 mg/dL, 95% CI (0.65, 6.93) (p=0.019). Treatment effect for HighCal: ΔBMD : 0.03 (p=0.16); ΔLDL, ΔTC and ΔTG: −25.41 (p=0.11), −22.14(p=0.07) and −2.65(p=0.64), respectively. LowCal: ΔBMD : −0.04 (p=0.16); ΔLDL, ΔTC and ΔTG: −10.86 (p=0.11), −5.96(p=0.07) and 4.53(p=0.64), respectively. ΔBMD was correlated with ΔLDL and ΔTC : −0.27377 (p=0.0519) and −0.27133 (p=0.0541), respectively.

Conclusion

Observed decrease in BMI, LDL, and TC; and increase in HDL. There was a weak correlation between Δ BMD, LDL and TC.

Sponsor: Loma Linda University
The relationship between life style factors and BMD in men from the Fourth KNHANES
Authors: Lee Okhee , Lee Jungyoon
Published: June 2021

Bone mineral density for adults were measured in the Fourth KNHANES. We evaluated the relationship of life style factors with bone mineral density of total femur, femoral neck and lumbar spine in Korean middle‐aged men. The used life style factors were body weight, dietary pattern, nutrient intake, marital status, smoking, alcohol drink, exercise habits. Their relationships with BMDs in men with age of 40–65y were examined by regression analysis. Foods and nutrients intake were used for analysis to energy intake by the residual method. Two dietary pattens were found by factor analysis and labeled as ˈhealthy Korean dietary patternˈ and ¡® unhealthy dietary pattern including many animal foodsˈ. Age, body weight, marital status, early age smoking, high intensive exercise explained the variance of BMD of total femur. The variances of BMD in femoral neck could be explained by age, body weight, marital status, high intensive exercise. In contrast, the BMD of lumbar spine could be explained by body weight and P intake. In conclusion, two dietary patterns had no relationships with bone mineral densities of femur and lumbar spine. Also, to maintain healthy femur, regularly exercise of high intensity were essential in middle aged men.
Prevalence and Determinants of Overweight and Obesity in a National Sample of 13–19 Year Old Lebanese Adolescents
Authors: Hwalla Nahla , Naja Farah , Sibai Abla , Adra Nada , Karam Sabine , Nasreddine Lara
Published: June 2021

The objectives of this study were to estimate the prevalence of overweight and obesity among Lebanese adolescents and to assess their socio‐demographic, lifestyle and dietary determinants. A nationally representative sample (N=446) of male and female 13–19 year old Lebanese adolescents were studied. Anthropometric, dietary, socio‐demographic variables were collected. The association between dietary patterns and overweight (including obesity) was also examined. Prevalence rates of obesity according to WHO 2007 standards were higher among males as compared to females (19.1% and 4.5%). Family history of obesity and sedentary levels were positively associated with overweight (OR=1.11) while female gender was negatively associated with overweight (OR=0.48). Higher odds of overweight were observed among adolescent boys with the highest intake of fat (OR=2.64) while lower odds were observed among adolescents with the highest intake of carbohydrates and fibers (OR= 0.50). Of the two dietary patterns identified (Western &Traditional), the Traditional was associated with lower odds of obesity (OR= 0.35). Findings of this study classify Lebanon among counties with high prevalence of male adolescent obesity. The identified determinants of overweight and obesity shed light on areas to be addressed and call for interventions and policy strategies to limit this epidemic in the country.
Nutrient Intake among Children with Autism
Authors: Moore Erin , Crook Tina , James Jill , Gonzales Dana , Hakkak Reza
Published: June 2021

Questions remain about potential nutrition deficits associated with autism. The purpose of this study was to determine if children with autistic disorder have adequate nutrient intake and if multivitamin use (MVIU) impacts intake. Participants (n=54) in the Autism IMAGE (Integrated Metabolic and Genomic Endeavor) Study at a local children's hospital completed Block Food Frequency Questionnaires (FFQ) for children ages 2 to 7. FFQ results were compared to Dietary Reference Intake (DRI) and Acceptable Macronutrient Distribution Ranges (AMDR). Mean intakes for % kcals from carbohydrate, protein and fat were within AMDR at 55% (SD=7), 14% (SD=3) and 33% (SD=5), respectively. However, intakes were below DRI for calcium (75% DRI ±45) and vitamins E (77% DRI ±26) and D (25% DRI ±19). A significant, positive relationship was found between MVIU and carbohydrate (p<.05); however, no significant relationships were found between MVIU and calcium, vitamins E or D, calories, protein or fat. Our results show that diets of children with autism are lacking in some micronutrients while macronutrient intake falls within recommended ranges.

Supported by 1Dept of Dietetics & Nutrition and 2LEND to JJ
Dietary patterns and hypertension among Chinese adults: a nationally representative cross‐sectional study
Authors: Wang Dong , He Yuna , Li Yanping , Luan Dechun , Yang Xiaoguang , Zhai Fengying , Ma Guansheng
Published: June 2021

Background

Little is known about the relationship between dietary patterns and hypertension among Chinese population.

Method

A nationally representative sample of 23 671 participants aged 18–59 years were recruited by the 2002 China National Nutrition and Health Survey. We conducted factor analysis using information from a validated food frequency questionnaire to derive dietary patterns.

Results

Three major dietary patterns, defined as ¡®Westernˈ, ¡®traditional northernˈ, and ¡®traditional southernˈ, were identified. T he score of the Western pattern is positively associated with blood pressure, whereas the score of the traditional southern pattern is negatively associated with blood pressure. In multivariate analyses the traditional northern pattern score was associated with an odds ratio (OR) of 1.30 (95% CI 1.11–1.53, P trend = 0.0001) comparing with the lowest quartile. The OR for the top quartile of score for the traditional southern pattern was 0.73 (95% CI, 0.59–0.89, P trend = 0.0040) compared with the lowest quartile of traditional southern pattern score.

Conclusion

We observed a strong positive relationship between the traditional northern pattern and hypertension. In addition, the traditional southern pattern was negatively associated with hypertension.
Caloric Titration Method (CTM) for weight loss in overweight and obese adults: 12 month results
Authors: Pacanowski Carly Rachael , Levitsky David A
Published: June 2021

The primary objective of this study is to experimentally test the efficacy of daily self weighing paired with a graphical representation of the individual's weight over time (versus no intervention) in producing a slow, clinically meaningful weight loss. Adults volunteered to participate in a weight loss trial (n = 178). Inclusion criteria consisted of having a BMI of at least 27.0kg/m2, absence of diabetes, pregnancy and eating disorder/history of eating disorder. Participants randomly assigned to the control group were tracked for the first year, and then given access to the intervention: frequent (ideally daily) weighing using a bathroom scale paired with an internet program designed to prompt 1% reductions in body weight by displaying a graphical representation of the user's weight. Despite lack of a statistically significant difference in average weight loss between groups (p = 0.10), 12 month analyses suggest that for some individuals, this intervention appears to be an effective self‐guided strategy for weight loss. Future research that identifies individuals that would benefit from this type of self‐directed approach is necessary.
Bioelectrical Impedance Analysis (BIA) and Anthropometric Data in Two Populations of Ecuadorian Children
Authors: Chukwueke Ihuoma Okezie , Beeson Lawrence W , Castillo Fritz , Cordero-MacIntyre Zaida
Published: June 2021

Objective

To make comparisons between Bioelectric Impedance Analysis (BIA) and anthropometry measurements in Ecuadorian children.

Methods

School age Ecuadorian children (171) between 7–15 years participated in the study. Body composition, fat free mass (FFM), and percent fat mass (PFM) were predicted from height, weight, skin fold thicknesses, circumferences (arms, waist, hips, and proximal thighs) and BIA measurements. FFM and PFM of subjects were evaluated by BIA‐Tanita (Model TBF 300A) and predicted using anthropometric equations.

Results

Significant correlations (Spearman's) were observed between Houtkooper's equation and BIA in measuring FFM (kg) (r=0.975, p=<0.0001) and between Slaughter's equation and BIA in measuring PFM (r=0.695, p=<0.0001) when combined for males and females. A Scatterplot for Houtkooper's equation vs. BIA for FFM fitted resulted in a regression line with a positive slope of 0.980 and a percentage variance of (Adj. R2=0.968). The Bland‐ Altman plots for Houtkooper's equation and BIA for FFM showed agreement between the two methods. A Scatter plot for Slaughter's anthropometric equation vs. BIA for PFM for both genders was fitted with a regression line with a positive slope of 0.778 resulted in a low percentage of variance of (adj. R2=0.559).

Conclusion

There were significant correlations between BIA and anthropometric measurements

Sponsor: Loma Linda University
Prepregnancy BMI and gestational weight gain as determinants of pregnancy complications and outcomes by race/ethnicity
Authors: Shin Dayeon , Song Won O.
Published: June 2021

Women of childbearing age have become race/ethnic diverse. We aimed to identify determinants of pregnancy outcomes by women's prepregnancy BMI (ppg BMI), gestational weight gain (GWG) and race/ethnicity in 3809 women aged 13–59y in NYC PRAMS 2004–07. Using SAS 9.1, logistic regression analyses were performed adjusting for age, marital status, education, income and gestational wks. The highest rate of preterm labor was seen in Hispanics (HP; 26.9%); infant ICU, pregnancy‐induced high blood pressure (HBP) and C‐section in African Americans (AA; 44.4%, 19.4%, 42.7%, resp.); and gestational diabetes (GDM) in Asian/Pacific Islanders (API; 18.7%). Compared to normal ppg BMI, obese ppg BMI had a higher rate (p<0.05) of GDM (OR=2.49), HBP (OR=1.88), and C‐section delivery (OR=1.37); whereas underweight ppg BMI had an OR=0.5 for HBP. Excessive GWG was associated with risks of HBP (OR=1.56) and C‐section (OR=1.34). The risk rates increased by the interaction of race/ethnicity, ppg BMI and GWG: infant ICU in API underweight ppg BMI (OR=2.44); preterm labor in AA obese ppg BMI (OR=0.65); and C‐section delivery in AA excessive GWG (OR=1.73); HBP (OR=2.08) and C‐section (OR=1.76) in non‐Hispanic White obese ppg BMI; and GDM in all race/ethnic obese ppg BMI. The general one set of GWG guidelines needs to be further evaluated to improve the health of mothers and infants of diverse population.

Grant Funding Source: None