bluuebunny/crossref_metadata_2025 huggingface dataset:
Histone deacetylase inhibition augments thyroid hormone (T3)‐induced increases in cardiac performance in hypothyroid rats
Authors: Plato Craig F , Joly Kristin M , Glascock Chris , Pitts Kelly R
Published: June 2021

Co‐administration of an HDAC inhibitor (HDACi) potentiates T3‐induced α‐MyHC expression in hypothyroid rats. We investigated the effects of a non‐selective HDACi, scriptaid (SA), on T3‐mediated increases in cardiac function in hypothyroid (HT) rats. Male Sprague‐Dawley rats were rendered rats HT by an iodine deficient, 0.15% propylthiouracil diet (2 wk). HT rats were treated (4d) with vehicle or T3 (3 μg/kg i.p.) and SA (1.5, 15 mg/kg i.p.) or its vehicle. Following treatment, cardiac performance and biochemical markers were determined and compared to euthyroid rats (ET). HT had low heart rate (HR; 235+6 bpm), decreased maximal +dP/dt (2755+209 mmHg/sec), increased tau (18+0.9 msec) and reduced α‐MyHC expression (88921 dlu) relative to ET (363+9 bpm; 6426+322 mmHg/sec; 9.0+0.9 msec, and 1058000 dlu, respectively). Although T3 elevated HR (330+8 bpm) and maximal +dP/dt (5614+421 mmHg/sec), reduced tau (12+0.7 msec), and upregulated α‐MyHC (963233 dlu) in HT, SA co‐administration significantly increased T3‐mediated increases in maximal +dP/dt (7234+471 mmHg/sec), reductions in tau (9.8+0.3 msec) and upregulation of α‐MyHC expression (1637000 dlu) without elevating circulating T3 concentration. Thus, SA augments T3‐induced increases in cardiac function and α‐MyHC expression in HT. These findings suggest HDACi exerts beneficial epigenetic effects on cardiac T3‐mediated transcriptional activity.
Effects of 20‐HETE analogues on intracellular signaling cascades in renal proximal tubular cells
Authors: Akbulut Talha , Regner Kevin R. , Roman Richard J. , Park Frank
Published: June 2021

We recently demonstrated that the 20‐hydroxyeicosatetraenoic acid (20‐HETE) analogues, 5,14‐20‐HEDE (WIT003) and 5,14‐20‐HEDGE (HEDGE) protect the kidney from ischemia‐reperfusion injury (IRI), and the protection is more pronounced with the HEDGE compound. In the present study, we performed in vitro assays using the LLC‐PK1 proximal tubular cell line to investigate the signaling cascades that may underlie the renoprotective effect of 20‐HETE analogues. Incubation with 20‐HETE (20 µM) for 60 min resulted in a rapid increase in the activation of the pro‐survival Erk1/2 pathway, including c‐raf (123 +/‐ 1%), MEK1/2 (285 +/‐ 8%), and ERK1/2 (493 +/‐ 36%) as determined by Western blot analysis. Similar increases in the ERK1/2 pathway were also detected using equimolar amounts of HEDGE and WIT003 with WIT003 being more efficacious than 20‐HETE and HEDGE (750% versus 20‐HETE and 500% versus HEDGE, p<0.001). Interestingly, WIT003 also activated the pro‐apoptotic SAPK/JNK (514%, p<0.0001) and p38 MAPK (411%, p=0.0001) pathways. In contrast, 20‐HETE and HEDGE did not activate these pathways. Our results suggest that HEDGE closely mimics the effects of 20‐HETE to selectively activate pro‐survival Erk1/2 pathway in LLC‐PK1 cells. WIT003, unlike HEDGE and 20‐HETE, also activates pro‐apoptotic pathways and this may explain why it is less effective at protecting the kidney following IRI in vivo.
Cardiac Microenviroment Program Human Aminotic Fluid‐derived Stem Cell To Cardiac Phenotype
Authors: Guan Xuan , Delo D. M. , Wang Z. , Soker Shay
Published: June 2021

Objective

It is believed that certain microenvironment can induce differentiation of stem cell into a specific cell lineage. Therefore, we carried out in vitro coculture experiments to investigate the potential of human amniotic fluid‐derived stem cell (hAFS) to differentiate along cardiac lineage. Our long‐term goal is to explore the feasibility of using this cell source for cardiac cell therapy.

Methods

Neonatal rat Cardiomyocytes (NRVM) were obtained by enzymatic digestion of ventricles from neonatal Sprague‐Dawley rats and plated on 1% gelatin coated coverslips. hAFS cells were labeled by PKH26, cultured directly with NRVM. At different time interval, cells were stained with cardiac specific antibodies. For dye transfer experiment, NRVM were preloaded by calcein‐AM and cocultured with PKH26 labeled hAFS.

Results

As early as 24 hours after initiation of coculture, contracting PKH26‐positive cells were observed. However, their number was low. After 14 days of coculture, immunostaining detected PKH26 positive cells colocalized with cardiac myosin heavy chain, cardiac Troponin T and connxin 43 positive cells. We further conducted dye transfer experiments to test coupling between NRVM and hAFS cells. 18 hour after initiation of coculture, calcein‐AM and PKH26 dual positive cells was observed, suggesting the existence of coupling between the two cell types, possibly through connexin 43.
Biofilm formation by biologic nanoparticles may require extracellular RNA and intact ribosomal function
Authors: Schwartz Maria A , Hunter Larry W , Lieske John C , Miller Virginia M
Published: June 2021

Biologic nanoparticles (NPs) isolated from human arterial calcifications and kidney stones form an adherent film on the bottom of flasks when placed in standard cell culture conditions. Since NPs have certain features consistent with microorganisms, we hypothesized that antibiotics and/or RNAse might inhibit this biofilm formation. NPs isolated from calcified human arterial tissue were cultured onto glass coverslips in 24‐well plates containing DMEM with 10% fetal calf serum, plus either no treatment (control), RNase, tetracycline or gentamicin. At 1, 7 and 14 days after inoculation, coverslips were fixed, mounted, and analyzed using a CytoViva darkfield scope and KS‐400 imaging software. In control wells, the percent coverslip covered by biofilm increased by 120% from day 1 to day 14, compared to 47% and 35% in wells treated with RNase and gentamicin, respectively. Wells treated with tetracycline showed a 3% decrease in biofilm by day 14. Since RNase should not enter intact cells, these results indicate that extracellular RNA may play a role in biofilm formation by NPs. Although tetracycline can chelate calcium, it also inhibits ribosomal function, as does gentamicin. Therefore these results also suggest that intact ribosomal function is required for NP‐associated biofilm growth. These experiments suggest that biofilm formation by NPs consists of biological as well as physicochemical processes.
MORPHOLOGICAL TYPES OF INTRAPARENCHYMAL SPATIAL DISTRIBUTION OF CAUDATE BRANCHES OF PORTAL HEPATIC VEIN. STUDY ON CORROSION CASTS
Authors: MATUSZ Petru L , HORDOVAN Eniko‐Christine , PUSZTAI Agneta Maria
Published: June 2021

Terminologia Anatomica (1998) homologates the caudate branches (CBr) of the portal hepatic vein (PHV) originating from the transverse portion of the left branch of the PHV. Our previous papers described the presence of CBr originating from other segments of the PHV. We analyzed the types of spatial distribution of CBr of the PHV according to their origin, on a number of 150 corrosion casts. The corrosion casts were made by injecting with plastic (AGO II paste and TECHNOVIT 7143) of the vasculo‐ductal systems, followed by parenchyma corrosion with hydrochloric acid. At the beginning we analyzed the branching modalities of the PHV trunk, finding three different aspects: Type I (modal, 97.33%), PHV forking into the right branch (RBr) and the left branch (LBr); Type II (2%), trifurcation of the PHV into LBr, anterior branch (ABr) and posterior branch (PBr); Type III (0.67%) bifurcation of PHV in LBr and PBr. On the study material we noticed the presence of 390 CBr, between 1‐6/cast. We found three morphological aspects of CBr: elongated (80%), forked (19.49%), in "bouquet" (0.51%). According to their number and origin, we found 23 morphological types of intraparenchymal spatial distribution of CBr of PHV. LBr is the origin of 75.39% CBr. We also found CBr from: RBr, trunk and bifurcation of the PHV, ABr and PBr. Knowledge about the CBr origin from other portions of the PHV than the transverse portion of the LBr is useful when planning and performing surgical approach of caudate lobe parenchyma (Supported by CNMP 41092/2007).
MORPHOLOGICAL TYPES OF INTRAPARENCHYMAL SPATIAL DISTRIBUTION OF CAUDATE LOBE ARTERIES. STUDY ON CORROSION CASTS
Authors: MATUSZ Petru L , HORDOVAN Eniko‐Christine , PUSZTAI Agneta Maria
Published: June 2021

Terminologia Anatomica (1998) homologates 2 arteries of the caudate lobe (CLA): right (RCLA) and left (LCLA), each originating from the homonym branch of the proper hepatic artery (PHA) (right branch ‐ RBr; left branch ‐ LBr). Our previous work evidenced variability both in origin and in number of CLA. We analyzed on 150 corrosion casts the types of spatial distribution of CLA according to their origin in different branching portions of PHA. The corrosion casts were made by injecting with plastic (AGO II paste and TECHNOVIT 7143) of the vasculo‐ductal systems, followed by parenchyma corrosion with hydrochloric acid. We analyzed at first the branching modalities of the PHA trunk and found 3 distinct aspects: Type I (modal, 74.67%) forking of PHA into right branch (RBr) and left branch (LBr); Type II (20.67%) forking of PHA into RBr and lateral branch (LBr); Type III (4.67%) forking of PHA in LBr and posterior branch (PBr). RBr is the origin of CLA in 33.67% cases, and LBr in 54.04%. We found origins of CLA from the anterior branch (1.05%), the posterior branch (1.40%), the lateral branch (2.81%) and the medial branch (7.02%). We found a higher frequency of origin of CLA from the medial branch in cases when it originates from the RBr. Evidence of CLA origin from other elements of the branching system of PHA than RBr and LBr is useful when planning and performing caudate lobe parenchyma surgery (Supported by CNMP 41092/2007).
Tendinous Architecture of Extensor Digitorum
Authors: Singh Keerti , Kesavi D. , Rajendran Melani
Published: June 2021

OBJECTIVE

To study the tendinous architecture of extensor digitorum during routine cadaveric dissections.

METHOD

Thirty (30) human cadavers were observed during routine dissections in the Department Of Anatomy of SRMC & RI, Chennai, India. The tendons of the extensor digitorum of these cadavers were examined on both sides for the number of extra slips. The origins and insertions of the extensor digitorum muscles were also observed.

RESULTS

Out of 30 cadavers, extra slips were observed in only thirteen (13) cadavers (43.3%). The adjacent tendons were connected by intertendinous slips.

CONCLUSIONS

The middle finger or the ring finger received doubled or tripled slips, which is a new finding. The index finger received only a single slip. No extra slip was found to the little finger or the thumb. The intertendinous connections were found to be normal as reported earlier. These accessory slips can be used in procedures of tendon rerouting by the plastic surgeons.

Grant Funding Source

University
The ligaments and articular surfaces of the talocalcaneal joint: a cadaveric study
Authors: Billard Kathryn , Padfield Beverly , Hrycyshyn Alan , Johnson Marjorie
Published: June 2021

Introduction

The articulation between the talus and the calcaneus, the subtalar joint, is important in chronic ankle instability. The relationship between the ligaments and the articular surfaces of this joint has not been thoroughly discussed in the literature. This study aims to describe and quantify the relationship between the morphology and attachments of the interosseous talocalcaneal ligament (ITCL) and the total surface area and configuration of the joint facets on the calcaneus.

Methods

20 subtalar joints from embalmed cadavers will be dissected and distracted. Measurements of the ITCL and the joint facets on the calcaneus will be reported.

Results

Preliminary results suggest a positive correlation between thickness of ITCL and total surface area of the talar facets on the calcaneus.

Discussion

Subtalar joint instability may contribute to recurrent ankle sprains. Knowledge of the ligaments and articular surfaces of the joint is critical for effective diagnosis and treatment. Reconstruction of the ITCL could be crucial in the treatment of chronic ankle instability. Precise information about the morphology, attachments and relationship of the ITCL to the joint facets is necessary to maximize the positive results of reconstructive surgery. This work was supported by the Anatomy & Cell Biology Department at the University of Western Ontario and the Ontario Graduate Scholarship program.

Grant Funding Source

OGS
Comparative histological methods for counting optic nerve fibers of the great horned owl, Bubo virginianus
Authors: Perkins Ashley Renee , Grady Shauna M , Walters Linda M , Suchocki Lawerence , Hall Margaret I
Published: June 2021

Optic nerve fibers are the axons of retinal ganglion cells, and as such they can provide information about the visual acuity and sensitivity capabilities of an animal. Currently, consistent optic nerve fiber counts are not available for most avians. Those that are available are dependent on method, with different methods yielding different counts. Here, optic nerve fibers are counted at light level for the Great Horned Owl. Juxtaposed sections of a single optic nerve are stained using three different techniques: Bielschowsky's silver impregnation, Bodian's protargol, and Luxol Fast Blue stain. The optic nerve is also observed using H&E to demonstrate details of microanatomy. We test two hypotheses: 1) different staining methods yield different myelinated optic nerve fiber counts due to variations in nerve fiber staining; and 2) hand counts and computer‐generated counts will vary depending on the stain. Counts are estimated for the nerve from 25 digital pictures taken at 1000X at regular intervals. Hand counts are performed multiple times by different observers and compared. Computer‐generated counts are performed in Image J. Research support comes from Midwestern University.

Grant Funding Source

AAA Annual Meeting Student/Postdoctoral & Young Faculty Travel Award Student Award
Attaching and Effacing Pathogen Infection Causes an Increase in Functional Connexin 43 Hemichannels to Generate Diarrhea.
Authors: Lin Ann En‐Ju , Li Yuling , Vogl A. Wayne , Bechberger John , Naus Christian , Finlay B. Brett , Guttman Julian A.
Published: June 2021

The attaching and effacing (A/E) pathogens enterohemorrhagic Escherichia coli and enteropathogenic E. coli cause serious diarrheal diseases. These organisms colonize the apices of the intestinal mucosa and inject pathogenic effectors into host cells altering sub‐cellular epithelial cell components. During these infections small molecules are released from the cells into the lumen of intestinal tracts, contributing to diarrhea. We investigate the role of gap junctions, a group of intercellular channels involved in the transport of small molecules and water, during A/E pathogen infection. Gap junctions are composed of paired connexon channels; however single connexon hemichannels also exist. Each connexon is formed by connexin (Cx) monomers. Using Citrobacter rodentium, a natural murine A/E pathogen, we show that Cx43 protein levels are increased and re‐localized during infection. Cx43 that normally localizes only to the lateral membrane of colonocytes was also localized at the infected cell apices forming functionally open hemichannels. During C. rodentium infection, Cx43 mutant mice had abolished diarrheal phenotypes while maintaining the bacterial colonization levels of wild‐type infections. These results provide the first evidence of connexon hemichannel formation to generate diarrhea and provides a novel target for future therapeutics to prevent bacterially‐induced diarrheal disease.

Grant Funding Source

CIHR and NSERC
Ability of n‐acetyl cysteine (NAC) to reverse TGF‐beta driven myofibroblast phenotype
Authors: Lu Daryn L , Vaughan Melville
Published: June 2021

The purpose of this study was to determine whether n‐acetyl cysteine (NAC) is able to reverse the Transforming Growth Factor‐β (TGF‐β) stimulated alpha‐smooth muscle actin (α‐sma) expression generated by myofibroblasts. Normal Carpal Tunnel fibroblasts and Dupuytren's diseased fibroblasts were grown on coverslips at a concentration of 0.5 x 105 cells per coverslip to allow sufficient time for growth to avoid quiescence. Three cell lines were used to help establish statistical significance along with one control.

This experiment was conducted in two phases. The first phase involved a control and a group treated with TGF‐β. All initial cell lines were grown in the presence of TGF‐β for an incubation period of 48 hours to stimulate the growth of myofibroblasts. The cells were then fixed and stained with immunofluorescence for a protein characteristic of α‐sma. After the initial incubation period, cells were harvested to determine how much TGF‐β increased α‐sma to establish a base line for future comparison. The second phase involved the group treated with TGF‐β to be split and treated with two additional treatments. One group continued with additional TGF‐β. The other group was treated with both TGF‐β and NAC to determine whether the base line increased, decreased or remained constant after an incubation time of 48 hours. After all cell groups were fixed and stained, the data was quantified and analyzed for statistical significance using an Analysis of Variance test (ANOVA).

It was expected that the group treated with additional TGF‐β would produce numerous quantities of myofibroblasts. It was also expected that the second group treated with both TGF‐β and NAC would remain constant without a significant increase in the myofibroblast population. Early results indicated NAC can reverse TGF‐β effects.

Grant Funding Source

University of Central Oklahoma Office of Research and Grants
System for large scale production of small RNAs through their in vivo expression within 5S rRNA‐derived scaffold.
Authors: Liu Yamei , Stepanov Victor , Dey Prasenjit , Jackson George W. , Willson Richard C. , Fox George E.
Published: June 2021

A stable system for manufacturing large quantity of small RNAs embedded into a 5S rRNA‐derived scaffold is under development. An RNA of interest will be incorporated into Bst aRNA, a deletion mutant of Vibrio proteolyticus 5S rRNA using standard molecular biological methods. The resulting chimeric RNA is expressed in Escherichia coli from either T7 or ribosomal P1/P2 promoters. The primary transcript is processed in vivo in the same way as endogenous 5S rRNA precursors but is not incorporated into E. coli ribosomes. As a result, the matured chimeric RNA accumulates to high levels in the bacterial cytoplasm. The RNA chimera can then be isolated, and the embedded RNA can be excised in vitro by appropriate enzymes. This system will provide a valuable and economic approach to produce defined small RNAs (such as siRNAs) in large quantity. Furthermore, the same system may be useful for the selection of novel small RNAs from a library of random RNA sequences incorporated into the 5S rRNA scaffold. The described approach will be of immediate interest in RNA pharmacological screening and drug development, and may ultimately find wider application.
Ribosomal frameshifting in response to hypomodified tRNAs in Xenopus oocytes
Authors: Carlson Bradley A , Lee Byeong Jae , Hatfield Dolph L
Published: June 2021

The question of whether hypomodified tRNAs enhanced mammalian retroviral ribosomal frameshifting (RF) was resolved several years ago when Phe tRNA lacking the highly modified wyebutoxine (Y) base at position 37 enhanced RF in retroviruses using UUU at the frameshift site (Carlson et al. Virology 255: 2, 1999). However, the level of RF in this earlier study and subsequent studies (Carlson et al. Virology 279: 130, 2001; Waas et al. JBC 282: 26026, 2007) far exceeded that naturally found with mammalian retroviruses which raised concerns whether Phe tRNA ‐Y only enhanced RF or was actually required for the event to occur. In the present study, we examined Xenopus oocytes as an "in vivo" protein synthesis system to assess whether the important issue of only an enhancement vs. an essential requirement could be resolved. Microinjection of oocytes with a construct encoding UUU at the frameshift site demonstrated that the level of RF was lower than that found naturally in retroviral RF, while co‐injection with Phe tRNA ‐Y enhanced RF and co‐injection with Phe tRNA +Y inhibited RF. Thus, Phe tRNA ‐Y does not only enhance RF, but is an essential requirement. This finding provides a novel avenue of inhibiting retroviral expression by reducing the level of intracellular hypomodified tRNA. This research was supported by the Intramural Research Program of the NIH, NCI, CCR.
The Effect of Chemerin on Glucose Homeostasis
Authors: Ernst Matthew C , Sinal Christopher J
Published: June 2021

Obesity, characterized by excess adipose mass, is a growing epidemic that increases the risk for developing cardiovascular disease and type 2 diabetes. Different mechanisms linking obesity with these comorbidities have been postulated, but remain poorly understood. Adipose tissue secretes a number of hormone‐like compounds, termed adipokines, that are important for the maintenance of normal glucose metabolism. Alterations in the secretion of adipokines are believed to contribute to the undesirable changes in glucose metabolism that commonly occur with obesity. Ultimately, this can result in the development of type 2 diabetes. We have identified a small secreted protein, chemerin, as a novel adipokine. Our study has shown that in mouse models of obesity, serum chemerin levels are significantly elevated and the expression of chemerin and its receptors, chemokine‐like receptor 1 and chemokine (C‐C motif) receptor‐like 2 are altered in white adipose, liver, pancreas, and skeletal muscle tissue. In addition, administration of exogenous chemerin exacerbates glucose intolerance and decreases serum insulin levels in obese mouse models. Therefore, chemerin has an important role in glucose homeostasis and may influence the metabolic derangements that result in obesity and type 2 diabetes. Funding: CIHR.
A cis‐proline in the hinge region of RNase A acts as a conformational gatekeeper for domain‐swapping
Authors: Karr Jessica , Miller Katherine , Marqusee Susan
Published: June 2021

Three‐dimensional domain swapping has been implicated in a wide range of functions including evolution of quaternary structure, activity regulation, and protein aggregation and amyloid formation. While the structural consequences of domain swapping have been well documented, little is understood about the specific structural and energetic features that cause this unique folding phenomenon to occur. An increased frequency of proline residues has been observed in hinge regions preceding swapped structural units, implying that the cis‐trans isomerization of proline may play an important role in domain swapping. For example, a trans proline may help extend the swapped arm, making it more accessible for exchange, while a cis‐proline may act as a conformational gatekeeper to prevent domain swapping by biasing the swapped arm towards contacts with the core of its own monomeric unit. Using RNase A as a model system, we have examined the role of a hinge region proline that undergoes cis to trans isomerization as the protein forms a domain‐swapped dimer. Results suggest that replacing the cis‐proline in the hinge region with a residue that energetically favors a trans configuration dramatically increases the ability of RNase A to form domain‐swapped dimers under physiological conditions.
Small scale modular programmable fermentation system.
Authors: Thatcher Chad , Lin Bae
Published: June 2021

A multi‐purpose, modular programmable fermentation system has been developed for growing a wide variety of microbial and cell types, including microbial, mammal, and plant cells. This small scale fermentation system provides a flexible platform for vessels from 0.5L to 15 L and can support single jacket vessels for microbial and double jacket vessels for mammalian cell culture. The system includes a 15 step programmable module, integrated control panel and software package which enables real‐time recording and execution of user's defined protocols. The controller allows keep the media conditions in the various growth phases for maximum yield, accommodating batch, fed batch and continuous fermentation as required. A pH probe enables automatic control of either acid or base media condition via peristaltic pumps and the DO probe controls the aerator. Additional ports are available to extend and customize control of the media environment. The small footprint, modular and upgradeable design and proven high reproductivity demonstrated by this system make it an ideal choice for various research projects required producing microorganisms and recombinant products.
Liver α2,6‐sialylation by ST6Gal‐1: Ablation of the hepatic promoter alters the α2,6‐sialylation pattern of serum glycoproteins but not the overall degree of sialylation
Authors: Migliore Marina Theresa , Jones Mark , Lau Joseph , Kazim Latif
Published: June 2021

The sialyltransferase ST6Gal‐1, which mediates the synthesis of the α2,6‐sialyl linkage to Gal( ?1,4)GlcNAc, is expressed ubiquitously but with particular abundance in the liver. The acute phase response, the liver's response to insult or injury, results in addition of heavily sialylated proteins into circulation. The acute phase response is also accompanied by hepatic up‐regulation of ST6Gal‐1 expression, but the functional relevance of hepatic ST6Gal‐1 remains unclear.

Previously, we generated a mutant mouse, Siat1ΔP1, by inactivation of the P1 region in the liver of the ST6Gal‐1 gene. Past experiments have shown myelopoetic disruption involving altered sialylation of serum glycoproteins. Initial examination demonstrated an essential unchanged degree of overall sialylation between wild‐type and δP1 animals, and serum glycoproteins in animals with globally inactivated ST6Gal‐1 remain fully sialylated. We use 2D gel electrophoresis, lectin affinity, and silver staining for analysis of serum glycoproteins to reveal significant qualitative and quantitative changes in the sialylation of serum glycoproteins between genotypes. These changes were further magnified after subcutaneous exposure of the animals to turpentine. We pay special attention to these mouse models at three crucial time points. Thus, the extent and types of sialyl linkages on serum glycoproteins are important.
Regulation of the Transcription Factor YY1 in Mitosis
Authors: Rizkallah Raed , Alexander Karen , Kassarjian Ari , Hurt Myra
Published: June 2021

YY1 is a multifunctional protein involved in the regulation of basic cellular processes at the DNA, RNA, and protein levels. The link between YY1 and cell cycle regulation has long been proposed. YY1 has been shown to interact with, and regulate, Rb, p53, c‐Myc, and many other cell cycle regulatory proteins. In addition, YY1 plays a role in the up‐regulation of many genes needed at the G2/M transition of the cell cycle. On the other hand, YY1 up‐regulates genes at the exit from mitosis, at the M/G1 transition. The subcellular localization of YY1 has been previously investigated at the G1/S transition of the cell cycle. In this study, the regulation and localization of YY1 during mitosis have been examined. Using confocal microscopy imaging of HeLa cells, YY1 was found to be primarily nuclear at the G2/M transition, but rapidly distributes to the cytoplasm, away from the condensed chromosomes, during prophase. YY1 remains excluded from DNA throughout pro‐metaphase, metaphase, and anaphase. At telophase, YY1 was rapidly recruited back to the decondensing chromosomes. The YY1 protein levels do not change significantly during the cell cycle; however, the YY1 protein in HeLa whole cell extracts was unable to bind its DNA consensus site in an in vitro electrophoretic mobility shift assay. The possibility of posttranslational modification of YY1 during mitosis is the focus of this study. Funded by FSU College of Medicine.
Determinants for stop‐transfer and post‐import pathways for protein targeting to the chloroplast inner envelope membrane
Authors: Viana Antonio Barbosa , Li Ming , Schnell Danny
Published: June 2021

Chloroplast biogenesis relies on the proper transport of thousands of nuclear encoded proteins into this organelle. The majority of nucleus‐encoded chloroplast proteins are imported via the TOC‐TIC translocons in the chloroplast envelope. Several of these proteins are further targeted to suborganellar compartments by additional targeting systems downstream of the import apparatus. The thylakoid targeting systems were presumably conserved from the original endosymbiont. In contrast, our knowledge of the mechanisms of protein targeting and integration at the inner envelope membrane (IM) is very limited. Previous reports have shown that IM proteins are transferred to the inner membrane via a stop‐transfer mechanism. Recent studies have demonstrated that one component of the Tic machinery is imported into the stroma and re‐inserted into the IM in a post‐import mechanism. This led us to investigate these pathways in more detail. Our analyses confirm the existence of both stop‐transfer and post‐import mechanisms of protein targeting to the IM. Furthermore, our results point to the conclusion that the transmembrane helices are not interchangeable and not sufficient for the choice of the insertion pathway. Further deletion and substitution analyses are being performed to better understand the features that determine protein insertion and topology into the chloroplast IM. Supported by NIH, CAPES/Fulbright.
Hexabromocyclododecane decreases the lytic function and ATP levels of human natural killer cells
Authors: Hinkson Natasha C , Whalen Margaret M
Published: June 2021

This study investigates the effect of Hexabromocyclododecane (HBCD) on human natural killer (NK) cell lytic function, additionally; it investigates the effects of HBCD on ATP levels in NK cells. Human NK cells are a subset of lymphocytes that are capable of lysing tumor cells, virally infected cells, and antibody‐coated cells. Hexabromocyclododecane (HBCD) is a non‐aromatic, brominated cyclic alkane used primarily as an additive flame retardant. If HBCD interferes with NK cell function, this could increase risk of tumor development and/or viral infection. NK cells were exposed to various concentrations of HBCD for 24 h, 48 h, and 6 days before determining lytic function and ATP levels. ATP levels and lytic function were also determined in NK cells that were exposed to HBCD for 1 h followed by 24 h, 48 h, and 6 days in HBCD‐free media. The results indicated that exposure of NK cells to 10 µM HBCD for 24 h causes a very significant decrease in both NK cell ATP levels and lytic function (90.5% and 93.5%, respectively). Exposure of NK cells to 10 µM HBCD for 1 h followed by 24h in HBCD‐free media also showed a very significant decrease in lytic function (89.4%) and a slight decrease in ATP levels (46.1%). The results indicate that HBCD exposures decrease ATP levels and lytic function in NK cells.

Supported by NIH grant SG06 GM008092‐33
'Rotation/twist' model for the EGF/ErbB receptor family activation
Authors: Maruyama Ichiro N.
Published: June 2021

The epidermal growth factor (EGF)/ErbB receptor family consists of four members, and plays pivotal roles in the development of organisms ranging from worms to humans. Furthermore, aberrant activation of the receptors is frequently implicated in a variety of human cancers. Ligand‐induced dimerization is proposed as a molecular mechanism underlying the activation of all the growth factor receptor tyrosine kinases including the ErbB receptor family. Before ligand binding, however, it remains controversial whether the receptors have a monomeric or dimeric structure. We have recently found that all the ErbB receptors have preformed, yet inactive, homo‐ and heterodimeric structures either on the cell surface or in the nucleus by bimolecular fluorescence complementation (BiFC). The spontaneous dimerization occurs in endoplasmic reticulum (ER), where ligands are unlikely to be available, before newly synthesized receptors reach the cell surface. These results are consistent with previous observations by in vivo chemical cross‐linking, cysteine disulfide bridging, FRET and FCCS. Based on current and previous results, we will propose the ?erotation/twist' model for the ErbB receptor family activation by ligand.
Isoangustone A isolated from hexane/ethanol extract of Glycyrrhiza uralensis induces apoptosis in DU145 human prostate cancer cells
Authors: Seon MiRa , Lim Soon Sung , Kim Jong‐Sang , Park Jung Han Yoon
Published: June 2021

Glycyrrhiza uralensis (licorice) is one of the most frequently prescribed agents in traditional herbal medicinal. We have previously observed that hexane/ethanol extract of Glycyrrhiza uralensis (HEGU) induced apoptosis and cell cycle arrest in DU145 human prostate cancer cell growth. This study examined the effects of the active compound isoangustone A in HEGU on apoptosis of DU145 cells. Isoangustone A (0 ‐ 7.5 mg/L) dose‐dependently reduced the viable cell numbers and increased apoptotic cell numbers in DU145 cells. Western blot analysis of total cell lysates revealed that isoangustone A increased the levels of Fas, TRAIL, cleaved caspase‐8 and Mcl‐1S and decreased the levels of Mcl‐1L. In addition, isoangustone A dose‐dependently increased the cleavage of poly(ADP‐ribose) polymerase and caspases 9, 7, and 3. We have demonstrated that isoangustone A inhibits DU145 cell growth via the induction of apoptosis.
Inhibition of fatty acid synthase by peptides from hydrolysates of β‐conglycinin enriched soybean
Authors: Roman Maxine , Martinez‐Villaluenga Cristina , Bringe Neal A , Gonzalez de Mejia Elvira
Published: June 2021

The synthesis of fatty acids is a target pathway for chemotherapy development because it has been linked to the growth and survival of cancer cells. The inhibition of fatty acid synthase (FAS) causes apoptosis of cancerous cells. Due to the chemical instability and negative side effects of existing FAS inhibitors, it is important to search for alternative inhibitory sources. The objectives of this study were to study the inhibitory effect of soybean hydrolysates (SH) derived from β‐conglycinin (BC) enriched soybean on FAS and characterize the responsible peptides. Total RNA from 3T3‐L1 adipocytes was isolated and FAS RNA expression was measured using a real‐time quantitative PCR method with SYBR green fluorescence dye. SH (100 μM) derived from soybean enriched BC (up to 47% total protein) showed a marked decrease of FAS mRNA abundance relative to the negative control. However, SH from 24% BC did not affect FAS gene expression (P < 0.0001, R2 = ‐0.90). It was concluded that soybean enriched in β‐conglycinin generates peptides that down regulate gene expression of enzymes involved in fatty acid synthesis in vitro, which can inhibit FAS. The potential health implications of this observation warrant further investigation.

Supported by The Monsanto Company, Cargill Co. and University of Illinois
Angiogenesis from endothelial progenitor cells is co‐regulated by vascular endothelial growth factor and matrix stiffness.
Authors: Hanjaya Putra Donny , Truitt Rachel , Yee Jane , Yee Derek , Ceci Doug , Gerecht Sharon
Published: June 2021

Angiogenesis from endothelial progenitor cells is co‐regulated by vascular endothelial growth factor and matrix stiffness. Endothelial Progenitor Cells (EPCs) in the circulatory system have been suggested to maintain vascular homeostatis and contribute to adult vascular regeneration. These processes require that EPCs break down the extracellular matrix (ECM), migrate, differentiate into endothelial cells, and line the lumen of the blood vessels. Evidently the ECM plays a critical role by providing biochemical and biophysical cues that regulate cellular behavior. Using our controllable in vitro model of angiogenesis we show that vascular endothelial growth factor (VEGF) and matrix stiffness regulate angiogenesis from EPCs. When exposed to low levels of VEGF, EPCs proliferate independently of matrix stiffness. Exposure to high VEGF concentrations results in upregulation of MMPs, demonstrating EPCs migratory and invasive abilities. Under these conditions, the stiffness of the matrix affects the progression of angiogenesis. Immunofluorescence and TEM images confirm the formation of vacuoles, which fully develop into lumen within a chain elongated cell on the softest matrix. RNAi studies further suggest that EPCs sense matrix stiffness through signaling cascades downstream of integrin ligation leading to the activation of the RhoGTPase and Cdc42. These results suggest that dual regulation of VEGF and matrix compliances are required to modulate angiogenesis dynamics of EPCs.

Grant Funding Source

AHA
Survey of biological active molecules in the Chinese traditional herbal formula Liu Wei Di Huang Wan for the treatment of Type II Diabetes
Authors: Mao Sena , Chuang G. , Tam W. , Tam G , Tam Chick
Published: June 2021

Objective

Identify biological active molecules in the ancient herbal formula Liu Wei Di Huang Wan (LWDHW) to treat diabetes in China.

Methods

Handbook of Active Constituents from Chinese Materia Medica & Phytomedicine (ISBN: 9620731549) by Wing and Gen.

Result

LWDHW (aka Rehmannia Six) is consist of six herbs: Rehmannia glutinosa (Shou Di Huang), Dioscorea opposita (Shan Yao), Poria cocos (Fu Ling), Paeonia moutan (Mu Dan), Cornus officinalis (Shan Zhu Yu) and Alisma plantago (Ze Xie). The emperor herb Rehmannia glutinosa contains jionoside A1 & B1, jiocarotenosides, four iridoid glucosides (catalpol, leonuride, aucubin, and melittoside); and four glycosides (rehmannioside A, B, C, and D) from the roots of species of var. purpurea and hueichingensis; glycosides (desrhamnosyl acteoside, forsythiaside, purpureaside A and B) from the leaves of species of Digitalis purpurea; and glycosides (purpureaside A & B & C, ateoside) from the callus of the same species.

LWDHW improves hyperglycemia and increases insulin sensitivity.

Conclusion

LWDHW has the bioactive function for the treatment of diabetes based on Traditional Chinese Medicine (TCM).
The effect of silicon supplementation on bone mineral density relative to Ca intake levels in ovariectomized rats
Authors: Kim Mi‐Hyun , Bae Yun‐Jung , Choi Mi‐Kyeong , Chung Yoon‐Sok
Published: June 2021

The purpose of this study was to investigate the effect of silicon (Si) supplementation on bone mineral density (BMD) and bone metabolism parameters relative to Ca intake levels in ovariectomized rats. A total of 72 female Wistar rats (six weeks) were ovariectomized (OVX) and divided into six groups, and Si (500 mg of Si per kg of feed) was or was not administered with diets containing various levels of Ca (0.1%, 0.5%, 1.5%) for ten weeks. Si supplementation significantly increased the BMD of the femur and tibia in Ca‐deficient OVX rats, while no change was observed with Si supplementation in the BMD of the spine, femur and tibia in the adequate and high Ca groups. Serum ALP and osteocalcin levels were not affected by Si supplementation or Ca intake levels. CTX levels were significantly decreased as a result of Si supplementation in Ca‐deficient OVX rats. In summary, Si supplementation produced positive effects on bone mineral density in Ca‐deficient OVX rats by reducing bone resorption. Therefore, Si supplementation may also prove to be helpful in preventing osteoporosis in postmenopausal women whose calcium intake is insufficient. <'This work was supported by the Korea Research Foundation Grant funded by the Korean Government (KRF‐2006‐331‐C00315)'>
Immunnostimulating and Anti‐stress Activities of Hot Water Extract from Pinus densiflora Sieb
Authors: Kim Jin‐Hwa , Joo Shin Youn , Kim Soon Im
Published: June 2021

This study performed to examine immunostimulating activities and anti‐stress activities of hot water extract from Pinus densiflora Sieb(PD). It is known that psychological stress affect the relationship between stress and immunological functions. This relationship has been revealed through complex interaction among the neurological, endocrine and immological systems. The immunostimulating activities were investigated with MTT assay by using human T cells, B cells and NK cells. The growth of human T cell was increased up to 109% in adding 100mg/l of hot water extract from PD and B cell was increased up to 107% at all concentrations. NK cell activation was significantly improved up 1.2 times higher than the case of control group. Anti‐stress effect of PD extracts was estimated by measuring blood chemical value and the internal organs weight in ICR mice. The forced swimming(5min) was given to animals as a stress factor. By these results, further studies on the product development using PD are required. This Research was financed by Seoul Research and Business Development Program(No. 10582), Republic of Korea.
Efficacy of a proposed binder against mycotoxins in rats.
Authors: Qiang Zhiyi , Myers Ronald K. , Hendrich Suzanne
Published: June 2021

A mycotoxin binder (MB) was hypothesized to decrease adverse effects of mycotoxins. 84 Sprague‐Dawley rats (21 day old) were fed control diet (AIN93G), MB (0.5% in diet), Toxins (12 ppm FB1, 8 ppm DON and 0.5 ppm ZEN), Toxins+MB or pairfed AIN93G equal to the Toxin‐fed group. Kidney wt in Toxin‐fed rats was decreased 5.1 % compared with controls (P<0.05), but Toxin+ MB‐fed rats did not differ from controls on d 15. Plasma alanine transaminase increased 48.6% and 27.9% in Toxin‐fed males and females, showing liver damage (P<0.05) compared with control group, and these FB1‐associated changes were prevented in rats fed Toxin+ MB. By HPLC analysis, the concentrations of urinary sphingosine (So) and sphinganine (Sa), biomarkers of FB1 toxicity, as Sa/So increased 11.3 and 12.0 fold in males or 11.6 and 10.4 fold in females fed the toxins (P<0.05) on both the 14th and 28th d compared with controls, with significantly less increase in males and females fed Toxin+MB (P<0.05) than in toxin fed animals on d 14. This lessened Sa/So ratio in rats fed Toxin + MB persisted at d 28 only in males. Histopathology analysis of tissues indicated that mild degenerative changes of tubular epithelial cells were found in males fed toxins, but they were not seen in the Toxin+MB group suggesting protection from toxic effects. Therefore, MB prevented or lessened effects of mycotoxins in rats. Funded by Kemin Nutrisurance, Des Moines, IA.
Transcriptomic signature of performance efficiency in finishing beef cattle
Authors: Burnett Derris D. , Kriese‐Anderson Lisa A , Wolfe Dwight F , Bergen Werner G
Published: June 2021

Metabolic redundancies and inefficiencies at the molecular level associated with feed efficiency (FE) and residual feed intake (RFI) of food animals may be ascertained by gene expression (GE) profiling. The objective of this study was to examine relationships between GE (mRNA abundance) of fatty acid synthase (FAS), PPARα, PPAR γ2, carnitine palmitoyl transferase (CPT‐1b), leptin, uncoupling protein 2 (UCP‐2), ubiquitin conjugating enzyme (E2) and polyubiquitin (PQ) genes in skeletal muscle (SKM) and adipose tissue (AT) with FE and RFI in finishing cattle. Crossbred cattle (bulls, 8; steers, 8) were placed on a feedlot diet and AT and SKM biopsies were taken prior to completion of the trial. Gene expression was determined with real time RT‐PCR. Adipose FAS, leptin and PPARγ2 GE were unrelated to RFI. SKM UCP‐2 and E‐2 but not CPT‐1b, PPARα and PQ GE were correlated with RFI (P<.05). AT leptin GE was higher (P<.05) while SKM UCP‐2 GE was lower (P<.05) in bulls than steers. While PQ GE was not correlated with RFI it was highly correlated (P<.01) with E2 GE. Results indicate that AT genes for fat deposition are likely minimally related to RFI and FE, while SKM GE associated with mitochondrial metabolism and protein turnover are correlated with RFI. The transcriptomic signature for efficiency in beef cattle appears more related to SKM than AT metabolism.
Beneficial immune effects in humans after regular consumption of juice from Concord grapes as determined by structural equation modeling
Authors: Rowe Cheryl A. , Nantz Meri P. , Nieves CJ , West Robin L. , Percival Susan S.
Published: June 2021

In a 9‐wk, randomized, double blind, placebo controlled human study, we measured several biological functions related to immunity. We previously reported that regular consumption of 100% grape juice made from Concord grapes resulted in increased γδ T cell numbers and higher serum vitamin C while consumption of a placebo juice resulted in reduced γδ T cell proliferation, lowered serum antioxidant capacity and increased DNA strand breaks in response to an oxidative stress. Structural equation modeling (SEM) was utilized to explore multiple relationships in the data. SEM is advantageous over traditional analyses in that it permits looking at an unobserved construct (in this case, biological function) which represents a multivariate combination of measures variables (in this case, the γδ T cell data, vitamin C, and DNA strand breaks). Our analysis shows that juice consumption was the strongest predictor of biological function at Time 2 (9 weeks), accounting for 61% of the variance. This analysis should be viewed as exploratory because it is, to our knowledge, the first attempt to test this type of model of biological function. Funded by Welch Foods, Inc. and the UF/IFAS Agriculture Experiment Station
Effect of diets high in fats or carbohydrates on the immune system of young BALB/c mice
Authors: Martínez‐Carrillo Beatriz E , Zúñiga‐Torres Guadalupe M. , Valdes‐Ramos Roxana , Rivera‐Aguilar Victor , Jarillo‐Luna Rosana A , Campos‐Rodríguez Rafael
Published: June 2021

Introduction

Increased dietary fat or carbohydrates affects metabolism, the immune system is no exception.

Objective

To evaluate the effect of dietary increase in fat or carbohydrate content on the immune system of a mouse model.

Methods

24 Balb/c mice were divided in three groups at 21 days of age. Control (C), high‐carbohydrate (HCO) or high fat (F) diets were fed ad libitum for 9 weeks. Intra‐ocular blood and total small intestines were obtained for analyses of lymphocytes using anti‐CD3+, CD4+ y CD8+ antibodies by flow‐cytometry.

Results

CD3+ cells were higher in the HCO (18.10% ±1.51) and F groups (17.31% ±1.53), than the C group (12.40% ± 1.42) (ANOVA=34.15 and <0.001). No differences were found in CD4+ cells between any of the groups. CD8+ cells were higher in the F group (14.83% ±1.42) compared with the C group (11.06% ±1.32), but not with the HCO group (12.42 ± 0.9) (ANOVA=258.23 and p< 0.001). Lamina propria CD3+ cells were lower in the F group (30.92% ± 3.62) when compared with HCO‐CD3+ (56.36 ± 6.76) and C‐CD3+ (44.34 ± 3.84) (ANOVA=83.38, p<0.001). F‐CD4+ (1.48% ±0.333) and F‐CD8+ (3.46% ±1.87) were lower the C‐CD4+ (19.98 ± 3.59), C‐CD8+ (22.29 ± 3.82), HCO‐CD4+ (20.24 ± 4.5) and HCO‐CD8+ (26.93 ± 4.6) (CD4+ ANOVA=52.78, p<0.001; CD8+ ANOVA=92.71, p<0.001).

Conclusions

T‐lymphocytes are decreased in peripheral blood by high carbohydrate or low fat diets, and decreased in the small intestine by a diet high in fat.