bluuebunny/crossref_metadata_2025 huggingface dataset:
Hyperoxia enhances response to respiratory syncytial virus (RSV) infection
Authors: Gladwell Wesley , Middleton Lance , Cook Donald , Kleeberger1 Steven R. , Ciencewicki Jon
Published: June 2021

RSV infects nearly all children by age 2, and typically causes mild host symptoms. Susceptible individuals develop severe lower respiratory tract infections and bronchiolitis, often leading to hospitalization or death. Clinical treatment often requires therapeutic oxygen. Previous studies showed that hyperoxia (>;95% O2) causes up‐regulation of TLR4, which has been implicated in host susceptibility and response to RSV infection. We hypothesized that hyperoxia enhances susceptibility and response to RSV infection in bronchial epithelial cells and inbred mice, and this effect is mediated by TLR4. BEAS‐2B cells were exposed to hyperoxia 18 hr and then infected with RSV. Cells were analyzed for markers of viral infection and pattern recognition receptors. Compared to air, hyperoxia caused a 4‐fold increase in viral load and increased TLR4 expression in cells 48 and 72 hr PI. C57BL/6J (WT) and C57BL/6J‐TLR4−/− mice were exposed to hyperoxia or air for 24 hr and 24 hr later were infected with a single dose of RSV or Hep 2.Viral load peaked 5 d PI in both strains however RSV in WT mice was significantly higher compared to TLR4−/− 1–5 d PI. Results indicate that hyperoxia enhanced susceptibility to RSV infection and exacerbated the resulting response, and this effect is partially mediated through TLR4. Support by NIEHS intramural research program.
Immunohistochemical Changes in 5 Respiratory Nuclei after Bilateral Carotid Body Denervation (CBD) in Sprague Dawley Rats
Authors: Mouradian Gary C , Forster Bert V , Hodges Matthew R
Published: June 2021

Bilateral CBD in rat has no effect on CO2 sensitivity but elicits hypoventilation 1–2 d post‐CBD followed by a return to baseline values 7–8 d post‐CBD. To investigate the neural mechanisms driving the recovery of breathing after CBD, we compared NK1 receptor staining intensity on neuronal membranes from 5 different respiratory nuclei (solitary tract, NTS; dorsal motor, DMV; hypoglossal, nXII; nucleus ambiguous, NA; pre‐botzinger complex, PBC). Brain tissue was obtained from sham (n=1) and CBD animals sacrificed 4–5 (n=1), and >;21 d (n=2) post‐CBD. Optical density (OD) measurements of NK1 in NTS, DMV, and nXII in the sham rat (0.91, 0.96, 0.80 OD, respectively) were similar to rats sacrificed after 21 d post CBD (0.84, 0.96, 0.74 OD, respectively). The rat sacrificed 4–5 d post‐CBD had less intense NK1 staining in the NTS (0.68 OD), DMV (0.77 OD), and nXII (0.65 OD) relative to the other group of rats. The sham rat had lower OD values in the NA (0.54 OD) and PBC (0.70 OD) than rats sacrificed after 21 d post‐CBD (0.93 and 0.93 OD, respectively). These preliminary data suggest that CBD causes a down‐regulation, and recovery from CBD due to up‐regulation of the NK1 receptor. Remaining tissue sections from the same animals will be used to compare AMPA type 2, NMDA type 1, the serotonin transporter, and 5HT2A receptor staining intensities in the same respiratory nuclei. Supported by NIH HL097033
A History of Giving Back and Caring: The Mount Sinai Tradition
Authors: Reidenberg Joy S. , Laitman Benjamin M. , Laitman Jeffrey T.
Published: June 2021

The Mount Sinai Hospital and School of Medicine has a long history of sharing knowledge with our neighboring community. In the past, medical education was considered a right and a privilege limited to the health professions. However, no career should have a monopoly on knowledge of the body. As scientist‐educators, our mission is to generate and disseminate knowledge. Outreach education to the lay community is a natural extension of that obligation. Our Center for Anatomy and Functional Morphology has spearheaded several educational programs that bring the community to the medical school for learning opportunities specifically focused on anatomy. Demonstrations are performed in our anatomy laboratory, where human cadaveric material can be safely displayed and touched by gloved hands. The experience is particularly memorable because the material is real, and the learning is kinesthetic. Students never forget the experience of holding a real human brain or heart in their hands. Viewing diseased organs successfully compliments and reinforces behavioral modification lessons taught in their school's curriculum, such as the harm inflicted by tobacco exposure or alcohol abuse.
Dietary Fiber Content and Benefit Labeling Did Not Increase High‐Fiber Bread Sales
Authors: Camire Mary Ellen , Morin Heidi England , Teisl Mario
Published: June 2021

Adolescent and young adult Americans generally do not consumer recommended amounts of dietary fiber (hereafter, fiber). This study attempts to increase sales of high‐fiber breads and wraps at a university food court by providing consumers information about fiber content, recommended servings, and the health benefits of fiber. IRB approval was obtained. Prior to the information treatment, 114 consumers aged 18–27 years completed a survey of fiber knowledge; a similar survey was given to 104 consumers after the treatment. For 2 weeks posters showed fiber content (grams/serving) of breads and wraps. After a 1‐week washout period, posters displayed fiber Dietary Reference Intakes (DRI), sources of fiber, and health benefits of fiber consumption were posted for 2‐weeks. Sales of the foods before, during, and after the interventions were estimated using dining services’ invoices. Sales of breads and wraps with higher (≥3 g/serving) fiber were not influenced by either type of labeling during the study period. Knowledge of fiber DRI was higher for women and for post‐intervention survey respondents. Other types of behavioral interventions may be needed to increase dietary fiber intake by young adults.
Anticancer effects of Garcinol in pancreatic cancer transgenic mouse model
Authors: Saadat Nadia , Akhtar Sarah , Gill Aminder , Razalli Nurul , Vemuri Shilpa , Goja Arvind , Geamanu Andreea , David Doina , Gupta Smiti
Published: June 2021

Poor prognosis in Pancreatic cancer (PC) is attributable to weak diagnostic and treatment options. Surgical resection is possible for 20% of diagnosed cases while current chemotherapy (Gemcitabine) has a 15% response rate. Previously, we reported that Garcinol, a polyisoprenylated benzophenone, from Garcinia indica, shows a favorable dose dependant response in PC cells, alone and in combination with Gemcitabine. In this study we report the invivo effects of dietary Garcinol in transgenic pancreatic cancer mouse model (KPC).

Methods

KPC (k‐ras and p53 conditional mutant) mice were divided into 4 groups: KC‐cancer control; KGr− 0.05% Garcinol diet; KGm‐Gemcitabine injected; KGG− Garcinol+Gemcitabine. Littermates without KPC mutations served as controls. PC progression was monitored by MRI and confirmed by tissues stained with Hematoxylin and Eosin. Blood smears were probed for changes in lymphocytes.

Results

The KGr group showed improved survival rate. MRI scans showed reduction in tumor volume in KGr, KGm and KGG groups. This was supported by the reduced number of Pancreatic Intraepithelial Neoplasia 3 (PanIN 3) in KGr and KGG groups as compared to the KC group. In addition, an increase in NK (natural killer) cells was observed in KGr group.

Taken together, our data indicates that dietary Garcinol decreased pancreatic cancer progression in transgenic pancreatic cancer mice.

Funded by Intramural funds.

Grant Funding Source: Intramural funds
The Effects of Behaviorally‐Based Diet Education and Tai‐Chi on Dietary Intake and Resiliency in Community Dwelling Older Adults
Authors: Taetzsch Amy , Lobuono Dara , Beebe Nowen , Letendre Jonathan , Bekke Jillian , Delmonico Matthew , Xu Furong , Lofgren Ingrid
Published: June 2021

Resilience is the ability to recover after a physical, emotional, financial, or social challenge. Limited research exists on the impact of dietary and physical activity interventions on resiliency. Fifty‐four obese, older women were recruited at two senior centers for a 16‐week community‐based intervention study that implemented a modified Dietary Approaches to Stop Hypertension (DASH) diet and Tai Chi (TC) exercise. At baseline, participants’ mean age was 68.2±7.7 years, with BMI of 35.4±4.3 kg/m2, and total cholesterol of 200.7±33.1 mg/dL. The mean Dietary Screening Tool (DST) score was 67.3±10.6 and the mean physical resiliency scale score was 12.96 ± 2.09. Significance was set at p<0.05. There was no significant difference at baseline between waitlist control (WC) and intervention (DASH‐TC) participants in any of the reported variables. One‐way repeated measures ANOVA comparing baseline to post‐intervention was significant for total DST score, BMI, waist circumference, systolic and diastolic blood pressure, and glucose; all with a large effect size. There was no significant interaction effect between WC and DASH‐TC groups in terms of physical resiliency; this may be related to a possible Hawthorne effect. The combination of DASH diet plus TC didn't seem to impact resiliency in this population. However, further research is needed to verify these results in larger, more diverse populations.
Dietary wolfberry up‐regulates BCDO2 and enhances mitochondrial biogenesis in the retina of db/db type 2 diabetic mice
Authors: Lin Dingbo , Ji Hua , Medeiros Denis M
Published: June 2021

Objective

To investigate whether wolfberry regulated β,β ‐ carotene 9′,10′‐dioxygenase 2 (BCDO2) expression and enhanced mitochondrial biogenesis in the diabetic retina.

Methods

Male db/db and wild‐type mice at 6 weeks of age were fed the control or wolfberry diets for 8 weeks. At study termination, the retina tissues were collected for analysis by transmission electron microscopy, real‐time PCR, immunoprecipitation, and Western blot.

Results

Wolfberry up‐regulated expression of retinal BCDO2 transcript and protein. Wolfberry induced activation and nuclear enrichment of retinal AMP‐activated protein kinase α2. Wolfberry attenuated hypoxia and mitochondrial stress by declined expression of hypoxia‐inducible factor‐1α, vascular endothelial growth factor, and heat shock protein 60. Wolfberry reversed mitochondrial dispersion in the retinal pigment epithelium, increased mitochondrial copy number, elevated citrate synthase activity, and upregulated the expression of peroxisome proliferator‐activated receptor γ co‐activator 1 α, nuclear respiratory factor 1, and mitochondrial transcription factor A.

Conclusions

Animal data suggest that increased consumption of dietary wolfberry enhances mitochondrial biogenesis leading to retinoprotection in diabetes. Grant Funding Source: National Institutes of Health COBRE Grant P20‐RR‐017686
Developmental expression and transcriptional regulation of claudin‐6 in the murine lung
Authors: Jimenez Felix R , Lewis Josh B , Wood Tyler T , Reynolds Paul R
Published: June 2021

Claudins contribute to tight junctions that prevent paracellular transport of extracellular fluid and diverse substances. Claudin‐6 was evaluated by qPCR and immunohistochemistry and its expression was confirmed to peak at E12.5 and diminish as development progressed. Co‐immunofluorescence revealed that Claudin‐6 co‐localized with Sox2 and Sox9, both factors that influence cell differentiation in the proximal and distal lung, respectively. Claudin‐6 was also detected in cells that express TTF‐1, a critical regulator of pulmonary branching morphogenesis. In order to assess transcriptional control of Claudin‐6, its promoter was ligated into a luciferase reporter vector and co‐transfected with TTF‐1 and other important transcriptional regulators. Factor‐specific effects were confirmed using site‐directed mutagenesis of putative response elements located in the Claudin‐6 promoter. This work revealed for the first time that Claudin‐6 might be an important tight junctional component during lung organogenesis. Furthermore, these and other data may reveal important Claudin‐6‐ mediated aspects of lung cell metastasis, remodeling, and disease states coincident with abnormal membrane permeability. This work was supported by a grant from the Flight Attendant's Medical Research Institute (FAMRI, PRR) and a BYU Mentoring Environment Grant (PRR).
In vivo imaging of cartilage and bone
Authors: Weinans Harrie
Published: June 2021

In‐vivo imaging in animal experiments has recently made enormous progression. Several clinical imaging techniques have been tuned to micro versions for small animal research with resolutions in the range of 20 microns for CT. Micro‐CT can be connected to other small animal imaging modalities such as luminescence and SPECT, thereby providing great opportunities for in‐vivo tracing of molecules, cells and their interactions. In research for skeletal diseases such as osteoarthritis, osteoporosis or bone regeneration a variety of mouse and rat models can be studied with in‐vivo micro‐CT thereby making sequential scans of individual animals at different time points in the disease or treatment process. Sometimes this is done in connection with other imaging techniques to further unravel the biological mechanisms. Micro‐CT can also be applied with contrast agents that diffuse in cartilage dependent on properties related to tissue quality such as e.g. the concentration of glycosaminoglycans. Altogether in‐vivo micro‐CT enables an enormous improvement in evaluating skeletal tissue in experimental animal models that triggers or speeds up the testing of new intervention strategies.
Polyphenols and phenolic acids modulate inflammatory cytokine release by Jurkat human CD4+ T‐cells
Authors: Ford Christopher Thomas , Richardson Sian , McArdle Francis , Crozier Alan , McArdle Anne , Jackson Malcolm J
Published: June 2021

Altered circulating levels of inflammatory cytokines are a risk factor for the development of age related diseases. The objective of this study was to assess the effect of polyphenols or phenolic acids on the release of cytokines from a human CD4+ T‐cell line (Jurkat cells). Cells were treated with a range of polyphenols or phenolic acids at 1 μM or 30 μM. Cells were treated with phorbol 12‐myristate 13‐acetate (PMA) stimulation after 24 hours and cytokine release was determined 24 hours later. Four compounds were identified that resulted in a significant reduction in interleukin 2 (IL‐2) and tumour necrosis factor α (TNFα) release by PMA‐stimulated Jurkat cells. These molecules were dihydroferulic acid (−56.5% TNFα, p<0.001), feruloylglycine (− 49.9% TNFα, p<0.001), quercetin (−67.2% IL‐2, p<0.001; −91.8% TNFα, p<0.001), and 3‐O‐methylquercetin (−91.5% TNFα, p<0.001). A further 3 compounds, epigallocatechin gallate (−46.2% IL‐8, p<0.01), punicalagin (−43.1% IL‐8, p<0.05), and callistephin (−48.5% IL‐8, p<0.01), reduced the release of cytokines by non PMA‐stimulated Jurkat cells. In conclusion, we have identified 7 dietary polyphenols or phenolic acids that reduced the release of inflammatory cytokines by a human CD4+ T‐cell line. The authors thank the British Biological Sciences Research Council Diet and Health Research Industry Club (BBSRC DRINC) programme for financial support.
Comparison of the bioaccessibility and Caco‐2 accumulation of lutein from human milk and infant formula
Authors: Lipkie Tristan E , Jouni Zeina E , Ferruzzi Mario G
Published: June 2021

In vivo studies in infants indicate that the bioavailability of lutein from infant formula is four times lower than from human milk. The purpose of this study was to assess the delivery of carotenoids from infant formula and human milk by using a coupled in vitro digestion and Caco‐2 human intestinal cell culture model. Thirteen human milk samples were assessed, including matched samples from three donors collected at 1, 3, and 6 months postpartum, as well as 10 infant formula prototypes with various formulations and carotenoid concentrations. The relative bioaccessibility of lutein did not differ (p>;0.05) between human milk (32±3%) and infant formula (30±3%), nor between lactation stages (p>;0.05). However, Caco‐2 uptake efficiency of lutein from digested media was 4 to 6‐fold greater (p<0.005) from digested human milk than from infant formula, and was significantly influenced by sample donor (p<0.05) but not by lactation stage (p>;0.05). While further studies are required to determine mechanisms involved, these data suggest the disparity between human milk and infant formula may be due to effects on intestinal absorptive processes, and not physical/chemical solubility or partitioning into mixed micelles. Funding provided by Mead Johnson Nutrition.
Hedgehog Signaling Pathway Regulates Autophagy in Human Hepatocellular Carcinoma Cells
Authors: Wang Ying , Han Chang , Wu Tong
Published: June 2021

Hedgehog (Hh) signaling plays an important role in the regulation of cellular functions. Classical Hh signaling is characterized by Smoothened (Smo)‐dependent activation of Gli1 and Gli2. Aberrant activation of Hh signaling has been implicated in several human cancers including hepatocellular carcinoma. In this study we examined the molecular mechanisms of Hh signaling pathway in HCC cells. Treatment of cultured human HCC cells with the Hh signaling ligand (recombinant Shh) or agonist, SAG and purmorphamine, prevented autophagy. In contrast, GANT61 (a small molecule inhibitor of Gli1 and Gli2) induced autophagy, as determined by immunobloting for microtubule‐associated protein light chain 3 (LC3) and p62, GFP‐LC3 puncta, monodansylcadaverine (MDC) staining and transmission electron microscopy. Hh inhibition‐induced autophagy was associated with Bnip3 upregulation. Knockdown of Bnip3 by RNAi impaired GANT61‐induced autophagy. Furthermore, inhibition of Hh signaling increased HCC cell apoptosis and decreased cell viability. The inhibition of autophagy by 3‐methyladenine (3‐MA) or Beclin‐1 siRNA partially suppressed GANT61‐induced cell apoptosis and cytotoxicity. In a tumor xenograft model using SCID mice inoculated with Huh7 cells, administration of GANT61 inhibited tumor formation and decreased tumor volume; this effect was partially blocked by the autophagy inhibitor, 3‐MA. These findings illustrate that hedgehog inhibition induces autophagy through upregulation of Bnip3 and this mechanism contributes to apoptosis.
A Common Docking (CD) Domain in Progesterone Receptor‐B Links Rapid Signaling Events to JAK/STAT‐dependent Gene Expression Required for Breast Cancer Cell Proliferation
Authors: Lange Carol Ann , Hagan Christy R
Published: June 2021

Progesterone receptors (PR) are context‐dependent transcription factors required for mammary gland development. PR target genes include paracrine factors that mediate mammary stem cell selfrenewal. In breast cancer cells, progestin‐bound PRs induce MAPK activation leading to regulation of growth‐promoting genes. Herein, we describe a common docking (CD) domain unique to PR‐B, a motif first described in MAPKs that facilitates direct interactions between MAPKs and MEKs or MAPK‐phosphatases (MKPs). Mutation of the PR‐B CD domain (mCD PR) did not alter progestin‐induced MAPK signaling. However, mCD PR failed to undergo phosphorylation on Ser81, a ck2‐ dependent site required for progestin‐regulation of growth‐promoting genes. PR Ser81 phosphorylation mapped to CD domain‐dependent binding of PR to MKP3. Phospho‐mutant S81A PRs failed to regulate PR‐B‐specific genes, including STAT5 and Wnt1, critical mediators of mammary stem cell expansion. Inhibition of JAK/STAT signaling blocked progestin‐induced STAT5 and Wnt1 expression. Our studies reveal a novel scaffolding action of MKP3 required for ck2‐dependent PR Ser81 phosphorylation. Co‐regulation of select target genes by phospho‐ Ser81 PR and STAT5 is likely a global mechanism required for the activation of growth promoting programs active during mammary gland development and relevant to mechanisms of breast cancer progression.
Student Concept Mapping: Conceptualization of Physiological Processes
Authors: Lee Karnatz Matthew , Ackerman Ralph A
Published: June 2021

We created an inquiry based concept mapping exercise for an upper level undergraduate physiology course with a lab component. We specifically focused on complex regulated systems (blood pressure, respiration, fluid balance, etc...) within physiology to promote conceptualization, not memorization. Students worked in small groups to determine what factors would be included and how they are related. Students utilized peer‐based feedback and multiple drafts to learn how to create a concept map. Assessment was accomplished by having students create a concept map to solve case studies or general disturbance problems. Once experienced at concept mapping, student groups were able to create a concept map to an unknown system within twenty minutes. This exercise can be used within a two hour long lab, or modified to a fifty minute lecture. Results are analyzed by comparing student performance to previous classes not using the inquiry based concept mapping approach.
Protein structure determination via X‐ray diffraction methods in an undergraduate biophysical laboratory course
Authors: Centeno Lani May , Dutnall Robert N.
Published: June 2021

Understanding the relationship between structure and function of biological macromolecules is a commonly emphasized concept in biochemistry and molecular biology. Teaching experimental aspects of structure determination is increasingly important so that students can navigate structure databases, understand method limitations, and critically interpret the growing body of structural information. Practical experience also has potential advantages to students starting graduate school or careers in industry. We sought to test the feasibility of undergraduate students in a laboratory class solving a protein structure, starting from purified protein, using X‐ray diffraction methods. We chose the serine protease trypsin as a target protein and used an in‐house X‐ray diffractometer recently acquired for teaching and research. We assessed strategies to grow crystals and collect high quality, high resolution (1.1 – 1.3 Å) data sets within a short time frame. Using the Phenix software program, we solved the structure via sulfur‐single wavelength anomalous diffraction and/or molecular replacement methods, and built and refined molecular models. Our results indicate that all aspects of this process can be implemented in a laboratory class. There is potential for altering the target protein to address hypothesis‐driven questions, and also for sharing data with other institutions for instructional purposes.
The effects of electrical stimulation and vibration therapy on the atrophic muscles of rats with spinal cord injury
Authors: Zamarioli Ariane , Butezloff Mariana , Issa João P , Maranho Daniel , Okubo Rodrigo , Falcai Mauricio , Shimano Antonio C
Published: June 2021

Spinal cord injury (SCI) often leads to a severe atrophy in the sublesional muscles. In this study we investigated the effects of electrical stimulation (ES) and vibration therapy (VT) on the magnitude of muscle loss in a rodent transection model of complete SCI. Young adult Wistar rats were divided into 4 groups: Sham, SCI, SCI+ES and SCI+VT. Both therapies initiated 3 days after SCI and were performed 3x/week, 20min for 30 days. Triceps surae were stimulated during ES (300‐μs rectangular pulses delivered at 50 Hz with a 5‐sec on/15‐sec off duty cycle), whereas stimulation amplitude was adjusted (20 to 150 mA) to produce a contraction that could be maintained for the duration of the stimulus. Rats were submitted to VT with frequency of 60Hz and vertical displacement of 1mm. On day 33 post‐SCI, gastrocnemius were dissected and analyzed by means of mechanical tests, morphology and histology. SCI significantly reduced cross‐sectional area, weight and mechanical resistance of muscles. We also observed that SCI caused a decrease of muscle volume and an increase of conjunctive tissue. ES showed efficacy at preventing the deleterious effects of SCI on muscle, by increasing cross‐sectional area, mechanical resistance, muscle fibers and by decreasing conjunctive tissue.
Endothelial Cell‐Dependent miR‐145 Expression Regulates TGFβ Signaling in Vascular Smooth Muscle Cells
Authors: Lilly Brenda , Zhao Ning
Published: June 2021

Blood vessel formation is a tightly regulated process that serves a critical role in both health and disease. Our lab has focused on the interactions between endothelial and smooth muscle cells that regulate proper blood vessel formation and function. In doing so, we identified miR‐145 as a microRNA that is robustly induced in vascular smooth muscle cells by endothelial cells. miR‐145 has been implicated in having an important role in the differentiation of smooth muscle cells. Our data show that miR‐145 is regulated by Notch signaling in smooth muscle cells, and when overexpressed leads to reduced migration. To investigate the underlying mechanisms, we searched for predicted targets of miR‐145 and identified TGFβ receptor II (TGFBR2) as a possible target. Overexpression of miR‐145 mimic and luciferase assays using the 3′‐UTR of TGFBR2 confirmed it is a direct target of miR‐145. Our results indicate that endothelial cell‐induced miR‐145 is regulated by Notch signaling, and once induced has the capacity to regulate TGFß signaling in smooth muscle cells. These data suggest that within smooth muscle cells miR‐145 acts to selectively regulate the TGFß pathway to govern smooth muscle function. Funded by AHA and Nationwide Children's Hospital.
Structure‐switching biosensors for the quantitative measurement of protein:protein interactions involved in aberrant transcriptional activity
Authors: Bonham Andrew J
Published: June 2021

Aberrant gene regulation is a key feature in a variety of human diseases, and this process is often initiated by altered proteinprotein interactions between different transcription factors. These physical associations can cause conformational changes or steric hindrance in the DNA‐binding domains of the transcription factors involved, altering site specificity and affinity. To better understand this process, we have developed sensitive, DNA‐based structure‐switching fluorescent biosensors for the quantitative measurement of the functional effects of these interactions on DNA‐binding activity. These biosensors allow convenient, quantitative measurement of both the kinetics and thermodynamics of DNA‐binding, and may have future academic and medical roles in assessing aberrant transcriptional regulation. Here, these biosensors were used to investigate the complex regulation process that occurs between the basic helix‐loop‐helix transcription factors Myc, Max, and Miz‐1. These transcription factors associate to form DNA‐binding complexes of varying affinity and specificity, and play key roles in cell growth and cancer progression. Ultimately, these biosensors may allow for greater insight into the process of oncogenesis and provide more reliable detection of the transcriptional regulation associated with early stage cancers.
A Single Gas Chromatography/Mass Spectrometry Method for Detection of Individual Enantiomers of a Broad Spectrum of G‐Type Chemical Warfare Nerve Agents
Authors: Kajih Takwen , Smith John , Rusek Tom , Kasten Shane , Cerasoli Douglas
Published: June 2021

The organophosphorus (OP) nerve agents are among the most toxic chemical compounds known. A subset of OP nerve agents known as G‐series compounds (including GA (tabun), GB (sarin), GD (soman) and GF (cyclosarin)) all share a chiral center at the phosphorus atom, resulting in a pair of stereoisomers designated as P(+) and P(−); the in vitro capacity to inhibit acetylcholinesterase as well as the in vivo toxicity of G‐agents is associated predominantly with the P(−) isomer of each compound. We have developed a gas chromatography/mass spectrometry (GC/MS) method to analyze the stereoisomers of all four G‐series nerve agents using a single chiral GC column. This approach allows successful baseline (or near baseline) resolution of each enantiomer of the G‐series agents under the same chromatographic conditions. Based on a variety of approaches, we have assigned each isomer peak as P(+) or P(−). The utility of the method (and variations thereof) was demonstrated by using this approach to determine the stereospecific rates of hydrolysis of GF as catalyzed by the enzyme OPH (from B. diminuta). The method described here will be generally applicable for screening enzymes to identify candidates that promote rapid hydrolysis of G‐agents, and thus may be effective as Bioscavengers of OP nerve agents.

Research support: Defense Threat Reduction Agency (DTRA).
Structural analysis of the GH10 xylanases Xyn10A and Xyn10C from Xanthomonas axonopodis pv. citri
Authors: Aadland Kelsey Solveig , Nong Guang , Preston James , Hurlbert Jason
Published: June 2021

Phytopathogenic species of Xanthomonas produce several glycoside hydrolases believed to contribute to pathogenesis by catalyzing the depolymerization of cell wall polysaccharides in susceptible hosts. The sequenced genomes of several Xanthomonas species have revealed two adjacent operons containing genetic orthologs to those encoding well‐characterized xylanolytic enzymes in other bacteria. In Xanthomonas axonopodis pv. citri, one of these operons was found to contain two genes encoding endoxylanases of glycoside hydrolase family 10 (GH10): xyn10A (xylanase 10A) and xyn10C (xylanase 10C). As part of a multidisciplinary and multi‐institutional effort to understand the role of these xylanases in the pathogenic process, we have begun structural analyses of the proteins encoded in this operon. We have cloned, expressed and purified Xyn10A and Xyn10C in Escherichia coli to sufficient purity to initiate crystallization trials. We have employed computational homology modeling with ab initio loop building to generate models of Xyn10A and Xyn10C that will be used in molecular replacement phasing of x‐ray diffraction data collected, thereby allowing us to determine the three‐dimensional structures of the proteins. This knowledge will be used to design protein‐specific inhibitors of the xylanases in order to abrogate the infectious abilities of X. axonopodis pv. citri.
Neuroprotection of estrogen of hypothalamic cell line due to hypoglycemic injury
Authors: Chakraborty Sanjoy
Published: June 2021

Glucose is the main metabolic fuel of the brain. Recurrent episodes of hypoglycemia can lead to seizures, coma, and even death. Hypothalamus involved in feeding and energy balance is most vulnerable to this injury. In the present study we report neuroprotection of estrogen on hypoglycemic injury in hypothalamic cell line. The cell viability, toxicity and proliferation were determined using different bioassays like cell count, MTT and LDH assay. There was a marked decrease in the cell count when exposed to hypoglycemia and the cell count increased in the presence of estrogen (p<0.0001). Under hypoglycemic conditions within 24 hours there was a significant loss in mitochondrial function with about 80% of retrieval in the presence of estrogen. The attenuation of the death pathways was only seen within the first 24 hours and gradually decreased with time. Cell damage and cellular toxicity was further analyzed by LDH. LDH increased in the absence of glucose and decreased in the presence of estrogen. In order to determine whether estrogen exerted its action through the AKT pathway, AKT activity was determined using PRAS40. There was reduction of PRAS40 when cells were exposed to hypoglycemic shock which dissipated in the presence of estrogen. Taking together the biochemical and molecular data it seems that estrogen shows neuroprotection against hypoglycemic shock and maybe via the activation of Akt pathway.
The Use of Pyrosequencing to Analyze Microbial Populations in Poultry Management to Minimize Downstream Food Contamination
Authors: Guttala Vamsy Priya , Bray Joey , Medrano Enrique Gino , Clack Beatrice A
Published: June 2021

The use of antibiotics in animals grown for human consumption has been banned in many parts of the world and is becoming less favorable in the US. With less or no antibiotics fed, the risk of microbial contamination of food is higher when processed. In the current study, the objective was to identify and compare the effect of both ceca and litter microbial communities present before and after treatment. The treatments consisted of 1) challenging newborn chicks at 3 days and then again at 38 days with either Salmonella enteritidis or Campylobacter jejuni or no bacteria in the presence of or without probiotics added to the feed and 2) windrow composting or no composting of litter as part of poultry house management. Aseptically ceca and litter samples were collected and stored at 4°C until processed. Bacterial genomic DNA was isolated from both sets of samples using either the Promega SV Wizard Genomic DNA Purification kit (Ceca) or Zymo‐spin soil microbe DNA midiprep kit (litter). The DNA from each sample was then amplified using 16s universal forward and reverse primers containing the specific Roche defined library‐L specific sequences as well as Multiplex Identifiers (MID) to allow for automated software identification of the samples after pooling, multiplexing and sequencing. We report the bioinformatic pipeline and the microbial distributions at the genus and species level for each treatment.. Pyrosequencing can be used to obtain valid comparisons between effects of various treatments on pathogenic versus beneficial bacteria in growth and management of animals used for human consumption.

Funding: Office of Research and Sponsored Projects, Stephen F. Austin State University; Ed and Gwen Cole, Nacogdoches, TX.
Nicotine increases the expression of alpha7‐nicotinic receptors (alpha7‐nAChRs) in human squamous cell lung cancer cells via Sp1/GATA pathway
Authors: Dasgupta Piyali , Lau Jamie K , Brown Kathleen C , Luo Haitao , Chen Yi C
Published: June 2021

Nicotine, the addictive component of cigarettes, promotes the proliferation of lung cancers via nicotinic acetylcholine receptors (nAChRs), specifically the α7‐nAChR subtype. We explore the effect of nicotine treatment on α7‐nAChR levels in squamous cell carcinomas of the lung (SCC‐L) in vitro and in vivo. Nicotine increased α7‐nAChR levels in three human SCC‐L cell lines. We also found that the levels of α7‐nAChR in SCC‐Ls (isolated from heavy smokers) were greater than the SCC‐L tumors isolated from moderate smokers. Nicotine‐induced up‐regulation of α7‐nAChR was dependent on α7‐nAChRs. The up‐regulation of α7‐nAChR upon nicotine treatment was confirmed in chicken chorioallantoic membranes (CAM) models. Real‐time PCR and luciferase assays showed that nicotine increased α7‐nAChR levels by transcriptional mechanisms in SCC‐L cells. ChIP showed that nicotine induced the binding of GATA4 or GATA6 to Sp1 on the α7‐nAChR‐promoter, thereby increasing its levels in H520 human SCC‐L cells. We also performed ChIP assay in SCC‐L tumors isolated from patients with heavy to moderate smoking history and obtained similar results. Long‐term exposure to nicotine could up‐regulate α7‐nAChRs on SCC‐L, thereby promoting its progression and metastasis. Our data would also be relevant to SCC‐L patients exposed to nicotine via second‐hand smoke or nicotine patches/gums used to quit smoking.
TCTP is Androgen‐Regulated and Over‐Expressed in Prostate Cancer
Authors: Storm Margrethe , Kaarbø Mari , Qu Su , Wæhre Håkon , Risberg Bjørn , Danielsen Håvard , Saatcioglu Fahri
Published: June 2021

TCTP has been implicated in a number of important cellular processes related to cell growth, cell cycle progression, malignant transformation and inhibition of apoptosis. In addition to these intracellular functions, TCTP has been shown to have extracellular functions and play an important role in immune cells. TCTP was previously shown to be expressed in prostate cancer cells, but its function in prostate cancer, or its relative expression compared to normal prostate, has not been assessed. Here we show that TCTP expression is regulated by androgens in prostate cancer cells in vitro and in vivo. Knockdown of TCTP reduced colony formation and increased apoptosis in LNCaP cells, implicating TCTP as an important factor for prostate cancer cell growth. In addition, LNCaP cells treated with recombinant TCTP increased colony formation, suggesting that secreted TCTP may function as a proliferative factor in the prostate. Furthermore, immunohistochemical analysis of human prostate cancer specimens showed that TCTP is over‐expressed in prostate cancer compared with normal prostate tissue. These results implicate TCTP as an important factor in prostate cancer both through inhibition of apoptosis and positively regulating cell proliferation.
Novel Strategy for Lineage Tracing of Cancer Stem Cells
Authors: Nguyen Tuan Minh , Chang Edmund C. , Rosen Jeffrey M.
Published: June 2021

The cancer stem cell (CSC) theory hypothesizes a subset of the tumorigenic cells able to self‐renew and wholly reconstitute original tumors by lineage restriction. We developed a lentiviral barcoding system as a means of genetic lineage tracing of cell subpopulations sorted by surface marker expression to investigate the proliferation kinetics, plasticity and clonality of tumor‐initiating cells (TICs). The abundance of each barcode in the reconstituted tumor reflects the frequency of TIC progeny in individual cell fractions. We determined that the CD29H/CD24H subfraction consistently out‐competes other subfractions while forming populations of different immune‐profiles, indicating that this subfraction retains the two cardinal features of self‐renewal and ability to give rise to progeny through lineage restriction ascribed to TICs, and suggests that not a lot of plasticity happens between non‐TICs and TICs. Also, iPCR was successfully applied into the lentiviral barcoding system, enabling the analysis of the TIC's clonality over serial transplant passages to understand the CSC's hierarchy. This novel lentiviral barcoding strategy provides future framework for defining specific biomarkers and evaluating therapeutics directed towards CSCs. This lineage tracing methodology should be applicable not only to genetically engineered mouse models but also to primary human breast cancer xenografts.
Use of HOMA indexes for the identification of metabolic syndrome and insulin resistance among Cuban‐Americans
Authors: Exebio Joel C , Ajabshir Sahar , Zarini Gustavo G , Shaban Lemia , Huffman Fatma G
Published: June 2021

Objective

To determine cut off points for HOMA‐1 and HOMA‐2 to identify insulin resistance and metabolic syndrome.

Methods

Subjects without diabetes residing in South Florida were enrolled (n=168, age 37 to 83 years). The HOMA‐1 and HOMA‐2 90th percentile in the healthy group (n=89) was used as the cut‐off point for insulin resistance. A ROC curve was constructed to determine the cut‐off point for metabolic syndrome.

Results

HOMA indexes were significantly correlated with fasting plasma glucose, central obesity, triglycerides and HDL cholesterol (P < 0.01). The cut‐off points for insulin resistance for HOMA‐1 and HOMA‐2 were >; 4.27 and >; 2.4 and for metabolic syndrome were >; 2.98 (69% sensitivity and 70.7% specificity) and >; 1.55 (65.4% sensitivity and 63.3% specificity), respectively.

Conclusion

HOMA cut‐off points may be used as a screening tool to identify insulin resistance and metabolic syndrome among Cuban‐Americans living in South Florida.
Maternal mid‐upper arm circumference (MUAC) is associated with lower birth weight among HIV‐exposed but not unexposed infants in a peri‐urban antenatal clinic in northwestern Tanzania
Authors: Wilkinson Amanda L , Pedersen Sarah H , Urassa Mark , Mkwashapi Denna M , Andreasen Aura , Todd Jim , Kinung'hi Safari , Changalucha John M , McDermid Joann M
Published: June 2021

Women in rural and semi‐urban areas of Africa face additional healthcare access barriers affecting birth outcomes and HIV‐exposed infants have a poorer prognosis compared to unexposed infants. Our objective was to characterize associations between maternal nutritional status during pregnancy and birth outcomes of HIV+ and HIV− women enrolled in a semi‐urban antenatal clinic cohort in northwestern Tanzania. Of the 46 HIV+ and 72 HIV−women enrolled, 21 HIV+ and 36 HIV− women delivered at/returned to clinic within 48 hours and were included in the analysis. Maternal anthropometrics were assessed at 20–36 weeks gestation, and HIV+ women were receiving antiretroviral therapy by delivery. Twenty nine percent of HIV+ and 22% of HIV−women had evidence of malnutrition (MUAC <25cm), but neither MUAC nor tricep skinfold (TSF) thickness differed by HIV status. HIV‐exposed neonates had lower birth weight and length compared with unexposed infants. Despite similarities in maternal anthropometrics, maternal MUAC and TSF predicted infant birth weight only in HIV‐exposed infants. Findings suggest that in HIV+ pregnant women living in rural or semi‐urban areas, maternal nutritional status based upon simple‐to‐measure anthropometry is an important determinant of infant anthropometrics among HIV‐exposed infants, and maternal MUAC identifies HIV+ women at higher risk of delivering low birth weight infants in this setting.

Grant Funding Source: Cornell University Unrestricted Grant
The proof is in the pudding: creation of an evidence‐based medical school nutritional education curriculum with cooking demonstrations
Authors: Joo Esther , Monlezun Dominique , Birkhead Andrew , Leong Benjamin , Abu‐Shamat Layla , Kay Danielle , Sarris Leah , Harlan Timothy
Published: June 2021

Inadequate nutritional education can result in what Vetter et al. (2008) demonstrated as only 14% of responding physicians reporting confidence in patient nutritional counseling. Wash et al. (2011) of Harvard Medical School report their integrated nutritional curriculum was unable to produce changes in medical student attitude and nutritional competence. Tulane School of Medicine recently responded to this growing need by creating its innovative medical student curriculum through the Tulane Center for Culinary Medicine. The curriculum includes pre‐session peer‐reviewed readings, lecture sessions, live cooking demonstrations, and clinical discussions. Medical students then translate clinically relevant culinary lessons into leading their own community‐based cooking demos. Curriculum validation is accomplished through longitudinal collection of participant surveys of competence and nutritional attitudes, correlated with their standardized board exams in their third and fourth clinical years, as well as their graduate medical education as residents. Research support: Tulane University Center for Culinary Medicine

Grant Funding Source: Tulane University Center for Culinary Medicine
Self regulation and diet as predictors of life satisfaction
Authors: Frye Kala M , Morrell Jesse Stabile , Carey Gale B , Warner Rebecca
Published: June 2021

This study examined whether self regulation predicts healthy diet and whether diet predicts life satisfaction among university students. Participants (n=333; 78% female) were recruited in Sept 2012 from the College Health and Nutrition Assessment Survey, an ongoing cross‐sectional study examining the health of traditionally‐aged (18–24 years) college adults at the University of New Hampshire. Data were collected from an online survey containing questions about self regulation skills, servings of fruits, vegetables and milk per day, servings of fish per week, and life satisfaction. Structural equation modeling was used to assess whether dietary variables may partially mediate the association between self regulation and life satisfaction. Statistically significant mediation was found only for fruits and vegetables; higher scores on self regulation predicted more daily fruit/vegetable consumption, and higher fruit/vegetable consumption predicted higher life satisfaction.
Role of Beta 3 Adrenergic Receptor in Facultative Thermogenesis
Authors: Preite Nailliw Zanini , Nascimento Bruna Pascarelli Pedrico , Lancellotti Carmen Lucia Penteado , Ribeiro Miriam Oliveira
Published: June 2021

Obesity, characterized by an increase in body fat, is related to a defect in facultative thermogenesis (FT). Brown adipose tissue (BAT) is the main site of this FT and is activated by Sympathetic Nervous System through the release of norepinephrine (NE) that binds to Beta adrenergic receptors (B‐ADR). Our goal was to evaluate the role of B3 isoform of B‐ADR in diet and cold‐induced FT. B3 knockout mice (KOB3) kept their body temperature normal when exposed to cold (4°C) and BAT thermogenic capacity was also normal when stimulated by NE i.v. infusion. In spite of a normal BAT activity, the absence of B3 receptor worsens diet‐induced obesity, with body weight gain significantly higher in KOB3 when compared to wild‐type (WT) mice (16g ± 1,4 vs. 12g ± 3,7) and larger epididymal adipocytes area (13079,81μm2 ± 10611,7 vs. 4501,55μm2 ± 1996,61). Also, a significant macrophages and neutrophils infiltration was observed in white adipose tissue (WAT), indicating inflammatory process in this tissue. Our results suggest that B3 receptor is not important in cold induced FT but it is fundamental to regulation of the body weight. It also indicates that this isoform mediates NE induced lipolysis since its absence results in very large adipocytes and inflammation in WAT.

Supported by CAPES.
Dermial®: A new natural ingredient with anti‐aging and regenerative properties for skin
Authors: Torrent Anna , Montell E. , Vergés J. , Ruhí R. , Ubía A. , Romero J. , Zurbano M.J.
Published: June 2021

Objective

To determine the anti‐aging and regenerative effect of Dermial®, a new ingredient with Cosmetic and Nutricosmetic potential.

Dermial® is a natural product rich in glycosaminoglycans, mainly hyaluronic acid, and also type I collagen, both beneficial for skin health.

Methods

The anti‐aging action was determined using a culture of human dermal fibroblasts (HDF). Skin hydration effect was quantified by measuring the synthesis of hyaluronic acid. The potential to increase skin elasticity was determined quantifying elastin levels. Effects on proliferation and migration of HDF were also tested.

Results

Dermial® showed a hydrating effect at the 3 tested concentrations in a dose‐dependent manner. This nutricosmetic ingredient was able to increase skin moisture up to 30% and to induce a statistical significant effect on the elastin synthesis up to 16%. In addition, this new ingredient was a potent inductor of fibroblast migration and was also able to increase up to 100% the proliferation index compared to the baseline culture.

Conclusions

These data demonstrate that Dermial® exhibits antiaging properties due to its hydrating action and its stimulating effect of elastin production. Moreover, it shows a powerful regenerative activity. All together, these results suggest that Dermial® may be useful as an anti‐aging treatment.

Supported by BIOIBERICA S.A., Barcelona, Spain.