bluuebunny/crossref_metadata_2025 huggingface dataset:
Phase I and II Clinical Trials on Recombinant H. pylori Vaccine
Authors: Zou Quan‐Ming , Zeng Ming , Zhu Feng‐cai , Mao Xu‐Hu , Tong Wen‐de , Guo Gang , Lu Dong‐Shui
Published: June 2021

Backgrounds: Phase I and II clinical trials on oral recombinant Hp vaccine were conducted from August 2003 to March 2004, in Yangzhong, Jiangsu. It is a recombinant Hp intramolecular adjuvant vaccine.

Methods: 653 healthy volunteers between age 6 and 15 that do not have Hp infection or Hp infection history were selected. All volunteers were clearly notified, and the experiment was random, double‐blinded, and controled. The volunteers were divided into 4 groups, which were given placebo, 15 mg, 30 mg, or 45 mg of vaccine per uptake. The vaccine and the placebo were taken orally on day 0, 14, and 28. Instant reaction was observed 30 min after each uptake. Thirty days of follow‐up visit were carried out, and abnormal reactions were monitored. At the same time, the levels of specific serum IgG, and total specific Ig of serum, along with specific sIgA of saliva and feces were detected before immunization, 14 days after the second immunization, 30 days, and 60 days after the third immunization.

Results: The vaccine is safe at all three doses, and no toxicity is observed. The levels of specific IgG, total specific Ig developed in serum were high in all three groups of subjects for a long period of time, and remained high 2 months after the last immunization. The antibody's positive seroconversion rates of the three groups of subjects, 15 mg, 30 mg, and 45 mg per uptake, were 95.6%, 97.8%, 92.3% respectively. However, the placebo group is only 3.8%.

Conclusions: This vaccine is safe and has strong immunogenicity. The effect of the vaccine is better after three immunizations than two immunizations.
Gamma‐irradiated pCD1‐ Yersinia pestis vaccine is protective: an anti‐LcrV response is not necessary to protect against the plague
Authors: Lambert Nathaniel Dane , Nilles Matthew L. , Bradley David S.
Published: June 2021

Yersinia pestis is the cause of plague and a potential weapon for bioterrorism. We have reported that immunization with lethally‐irradiated Y. pestis Kim5 is protective against subsequent i.v. challenges with the plague. We report here that pCD1‐Y. pestis, lacking the TTSS, provides protection comparable to wild‐type Y. pestis. Both strains provided protection against >107 CFU challenge of wildtype Y. pestis, compared to sham‐vaccinated mice that displayed an LD50 of 1.4X104 CFUs. This finding suggests that LcrV, located on pCD1 and previously shown to provide significant protection, was not essential for protection. A limited number of antibody specificities were generated by the irradiated wildtype or pCD1‐ Y. pestis, 6–8 unique specificities as determined by immunoblotting. The majority of these immunodominant specificities were shown to reside on the outer surface of the bacteria. Although this work has focused on humoral responses, T cell immunodominant epitopes can be rapidly identified with the irradiated vaccine as well. Lethal gamma‐irradiation provides a novel approach for the rapid production of vaccines that have remained elusive. Using a whole cell vaccine also allows for the rapid assessment of the immunodominant “footprint” to the majority of antigens present, aiding in the identification of putative sub‐unit vaccine candidates.
Carbohydrate mimetics as potential antiinfective treatments for Staphylococcus aureus
Authors: Fagan Diana L. , Royal Maurice , Vicarel Monica , Norris Peter
Published: June 2021

Antibiotic resistant strains of Staphylococcus aureus are an important cause of nosocomial infections. These strains are currently causing community‐acquired infections, resulting in an increased need for alternative therapies. S. aureus capsules appear to act as important virulence determinants, as bacteria with type 5 and 8 capsules cause 75–90% of nosocomial infections and anti‐capsular antibodies decrease mortality in infected mice. Our research involves construction of analogs of the aminosugars found in S. aureus capsules that we are testing for their ability to prevent capsule synthesis. To measure capsule production, bacteria were incubated with MV‐II‐065 or MV‐II‐063 (both n‐glucosyl 1,2,3‐triazoles with different functional groups), formalinized, trypsinized, and the bacteria used as an antigen in an ELISA. The presence of capsular carbohydrate was determined by incubation with anti‐capsular monoclonal antibodies, followed by a secondary incubation with peroxidase‐conjugate anti‐mouse Ig and a colorimetric substrate. Incubation type 5 S. Aureus with MV‐II‐065 decreased antibody binding, but the compound had no effect on Type 8 capsule synthesis. Studies with MV‐II‐063 (shown below) have indicated inhibition of capsule synthesis in Type 8 bacteria, while type 5 capsule synthesis remains to be tested.

In addition to their possible therapeutic potential, the differential effects of these two compounds may be used to investigate the synthetic pathways for these capsules. Support: NIH R15 2003‐2006, YSU PACER.
Dominant arginase expression in a model of progressive visceral leishmaniasis
Authors: Osorio Yaneth E. , Zhao Weiguo , Espitia Claudia , Travi Bruno L. , Hawel Leo , Byus Craig V. , Saldarriaga Omar A. , Melby Peter C.
Published: June 2021

Progressive disease in the hamster model of visceral leishmaniasis (VL), caused by Leishmania donovani, mimics human VL. During progressive infection in hamsters, there is low expression of NOS2 but high splenic arginase I mRNA expression (p=0.005) and enzyme activity (p=0.016), and its downstream products, polyamines (p<0.008). In contrast, the controlled infection in mice is accompanied by high NOS2 expression but low arginase activity and low polyamine production. Increased arginase activity and polyamine synthesis was also evident in an in vitro model of infected hamster macrophages (p<0.03). Leishmania arginase made little contribution to the overall increase in arginase activity. During progressive disease, there was increased STAT6‐inducing activity in the serum and spleen, and increased splenic expression of IL‐4, IL‐10, IL‐13, and IL‐21. Parasite‐induced macrophage arginase expression was amplified by exogenous IL‐4 and IL‐10, possibly through the infection‐induced upregulation of IL‐4 and IL‐10 receptors. The targeting of this pathway has therapeutic potential for this and other intracellular pathogens. Supported by NIAID.
NK cell stimulatory receptors and ligands are induced post‐transplantation
Authors: Wai Lu‐En , Fujiki Masato , Zhuo Ming , Martinez Olivia M. , Krams Sheri M.
Published: June 2021

Studies suggest a role for NK cells in acute and chronic allograft rejection. Further we have shown that calcineurin inhibitors do not block NK cell function. NK cells express inhibitory receptors that bind to classical MHC class I molecules and prevent killing. NK cells also express stimulatory receptors that activate and/or regulate effector function. In the context of transplantation NK cells are a major source of IFNγ early post‐transplant, however the expression and functional significance of NK cell receptors in the alloimmune response remain unclear.

Using an experimental model of rat liver transplantation we show by quantitative real‐time PCR (qPCR), that the stimulatory receptors NKp46 and NKG2D are increased in allografts, but not syngeneic grafts, by day 3 post‐transplant. Similarly, we have cloned two rat NKG2D ligands and demonstrate that their expression is silent in normal liver, induced upon liver transplantation, and specifically increased in allografts by day 3. In contrast, increased expression of the stimulatory receptor NKp30 is detected in allografts on the first day post‐transplant. Further, NKp30 mediates NK cell interaction with dendritic cells leading to abundant IFNγ production. Our data demonstrate the concomitant expression of NK receptors and induced ligands in allografts and suggests that NK cells are a functional link between innate and adaptive immunity post‐transplantation. Supported by NIH AI44095
Farnesyl transferase inhibitors delay primary skin allograft rejection in class II MHC‐mismatched mice
Authors: Gaylo Alison E. , Batzel Erika J. , Laux Kathleen S. , Field Ken A.
Published: June 2021

We are investigating the possibility that farnesyl transferase inhibitors (FTIs) may be useful anti‐rejection drugs. We have shown that the FTI, L‐744,832 can significantly delay allograft rejection in mice using MHC class II mismatched skin allografts. When treatment with this FTI begins two days prior to the allograft procedure and extends for 5 days, we found that rejection was delayed in all treated mice, 26±14 days, compared to untreated mice, 13±4 days. Interestingly, this delay in rejection can extend for greater than 30 days in 30% of the treated mice. To determine whether FTI treatment can delay alloreactive effector responses, we examined the effect of L‐744,832 treatment on allograft rejection in mice that had previously been sensitized to the allo‐antigens. We found that treatment with this FTI did not significantly delay second‐set allograft rejection. This observation, coupled with the long delay in some of the first set allograft treatments, suggests that FTIs may be able to induce tolerance of the alloreactive CD4+ T cells. To further investigate the action of FTIs on graft rejection we are measuring the effects of treatment using mixed lymphocyte cultures and using in vivo tolerance assays. We suggest that FTIs may modulate acute immune responses without systemically suppressing the immune system.
IgVH genes encoding xenoantibodies directed at GalTKO xenoantigens in rhesus monkeys
Authors: Harnden Ivan , Kiernan Kathleen , Gunthart Mirja , Gregory Clare , Meisner Jessica , Kearns‐Jonker Mary
Published: June 2021

Galactoslytransferase‐deficient (GalTKO) pig organs carry great potential as a solution to the shortage of human organ donors. These organs do not initiate hyperacute rejection but undergo acute humoral rejection mediated by xenoantibodies directed at non‐gal xenoantigens. The structure, specificity and germline origin of these antibodies has not been determined.

METHODS: Three rhesus monkeys were immunized with sixty million GalTKO pig endothelial cells in order to induce high levels of xenoantibodies directed at GalTKO xenoantigens. The specificity of the immune response to GalTKO xenoantigens was confirmed by flow cytometry. Pre and post transplant peripheral blood B cells were used to prepare IgG and IgM cDNA libraries for identification of the IgVH genes encoding xenoantibodies.

RESULTS: IgVH genes encoding anti‐non‐gal xenoantibodies were most closely related to the human germline progenitor V3‐21. The CDR3 region is unique and the J region is most similar to JH4 in 75.8% of the IgG sequences. The germline progenitors encoding anti‐non‐gal xenoantibodies are restricted and are 91% similar to the IGHV3‐11 alleles that encode anti‐carbohydrate xenoantibodies in primates transplanted with wild type and hDAF transgenic pig organs.

CONCLUSIONS: The anti‐non‐gal xenoantibody response is restricted to usage of an IgVH repertoire known to encode antibodies to carbohydrates and glycolipids.
Dictyostelium discoideum amoeboid cells respond to microbial ligands
Authors: Snyder Michelle Dykstra , Cassilly Daniel , Samaroo Dave , Walk Alex , Sanborn Keri
Published: June 2021

Mammalian innate immune cells detect pathogen‐associated molecular patterns (PAMPs) using evolutionarily‐conserved pattern‐recognition receptors that can be studied using a variety of model organisms. Dictyostelium discoideum is a unique model organism that exists for part of its lifecycle as single‐celled amoebae, but is induced to form a multicellular sporulating body upon starvation. As amoeboid cells, D. discoideum phagocytose bacteria for nutrient uptake; however, it has not been appreciated whether D. discoideum use pattern‐recognition receptors to detect bacterial prey. Here we show that D. discoideum cells indeed respond to known PAMPs, as exposure to the bacterial cell wall products LPS and peptidoglycan induces the production of reactive oxygen species. Furthermore, pretreatment of starving cells with LPS delays the development of sporulating bodies. Finally, as measured by qRT‐PCR, exposure of amoeboid cells to LPS and peptidoglycan upregulates the expression of genes homologous to those known to function in microbial recognition. Upregulated genes include a putative scavenger receptor and a gene containing the toll/interleukin‐1 receptor domain. We are currently creating D. discoideum cells that overexpress or are deficient for these gene products. The characterization of such genes should provide insight into conserved molecular mechanisms underlying microbial detection.
Acute effect of oral nutrients on fibrinogen synthesis in healthy young and elderly subjects
Authors: Caso Giuseppe , Maturi Jyothirmayi , Mileva Izolda , Kelly Patricia , Ahn Hongshik , Gelato Marie C , McNurlan Margaret A
Published: June 2021

The hepatic synthesis of fibrinogen is stimulated following inflammation and trauma. However, it is not known if fibrinogen synthesis is responsive to oral nutrients and, further, whether the meal effect may be influenced by age. The aim of the study was to assess the acute effect of oral feeding on fibrinogen synthesis, in both young and elderly subjects. Fibrinogen synthesis was determined in three separate occasions from the incorporation of L[2H5]phenylalanine (43 mg/kg) in 8 young (21–35 y) and 6 elderly (>60 y) subjects following the ingestion of water (control), a complete liquid meal (15% protein, 30% fat and 55% CHO), or only the protein component of the meal. The ingestion of the complete meal significantly enhanced fibrinogen fractional synthesis rates (FSR) in both groups by 30% (P < 0.005). A comparable stimulation of FSR was observed with only the protein component of the meal in both young (+36 %) and elderly subjects (+29 %) (P<0.001). Similar results were obtained when fibrinogen synthesis was expressed as absolute synthesis rates (ASR, i.e. mg/kg/day). The results demonstrate that fibrinogen synthesis is acutely enhanced after feeding and that this effect can be elicited by the protein component of the meal alone, both in young and elderly subjects.

Supported by NIH grants AG 017446 and 5‐MO1‐RR‐10710 and the NY State Empire Clinical Investigator Program (ECRIP)
Citrulline directly modulates muscle protein synthesis: evidence from in vitro study
Authors: Moinard Christophe , Jourdan Marion , Walrand Stéphane , Le Plenier Servane , Boirie Yves , Cynober Luc
Published: June 2021

Introduction : The ability of citrulline (CIT) to stimulate muscle protein synthesis has been recently reported. The question of how CIT can stimulate muscle protein synthesis thus arises. The aim of this work is to evaluate the direct effect of CIT on muscle protein synthesis rate using a model of incubated skeletal muscle. The study was performed in healthy and malnourished rats.

Material and methods : 3 month‐old rats were submitted to dietary restriction (50 % of the spontaneous food intake with a 5% protein diet) during 6 weeks (R, n=10) or fed ad libitum(AL, n=10). Epitrochlearismuscles were removed and were placed in a tube containing 3mL of appropriate medium (Krebs‐Ringer). Muscles were first incubated for 30 min at 37°C with 95%O2:5%CO2. Muscles were then incubated for 2 hours in medium containing insulin (0.01 U/ml) and [ring‐13C6] L‐phenylalanine (1mM) with (left) or without (right) CIT (2.5 mM) under the conditions described above. At the end of the incubation period, muscles and media were frozen and stored at −80°C. The absolute synthesis rate (ASR) was determined using the [ring‐13C6]‐phe enrichment of intracellular free amino acids and proteins by mass spectrometry.

Results:

Conclusions: this study demonstrates for the first time that CIT has a direct effect on muscle protein synthesis and that action is not related to the generation of other amino acids.

This work was supported by funds from the Ministry of Research and Technology
Post‐exercise meal timing alleviates decreased protein efficiency for skeletal muscle growth in rats fed high‐protein diet
Authors: Kondo Emi , Inoue Nagisa , Okada Sachiko , Yaji Keiko , Hirota Ayumi , Okamura Koji
Published: June 2021

To examine whether the upper limit of dietary protein for skeletal muscle growth would be increased if protein is ingested in proper timing in relation to exercise, rats fed at 8:00–10:00 and 20:00–22:00 were exercised with a climbing cage (Kondo et al. EB2007), 3 times a week for 8 weeks. The exercise started at 14:00 (14E) or 18:00 (18E). The proximate meal located 4h after exercise for 14E and immediately after exercise for 18E. The rats were fed either a normal protein diet (N, protein 18%) or a high protein diet (H, protein 35%), and each diet group was divided to a sedentary (S), 14E or 18E group. The rats were pair‐weighted. Nitrogen balance measured in the 7th week of the study was more positive in the H‐fed groups than the N‐fed groups without differences regarding both meal timing and exercise. After 8 weeks, the skeletal muscle mass (sum of the gastrocnemius, soleus, plantaris, Extensor digitorum longus, tibialis anterior, flexor hallucis longus and biceps brachii) was lower in the H‐fed rats than the N‐fed rats, and it was lower in 14EH (8.29g (SD 0.63)) than both SH (8.99 (0.38)) and 18EH (9.22 (0.53)) while SH and 18EH were comparable. The difference in the skeletal muscle mass between H‐fed and N‐fed was less in 18E than both S and 14E (SN > SH (−3.1%), 14EN > 14EH (−7.7%), 18EN = 18EH (−0.8%)). These results suggest that ingesting a meal in proper timing after exercise may not increase the upper limit of dietary protein for skeletal muscle growth, while it appears to alleviate the decreased efficiency of skeletal muscle growth in H‐fed animals.
Tissue‐specific requirement for TGF‐β signaling during craniofacial development
Authors: Chai Yang
Published: June 2021

Members of the transforming growth factor‐β (TGF‐β) superfamily mediate a wide range of biological activities, including cell proliferation, differentiation, extracellular matrix formation, and induction of homeobox genes, suggesting that TGF‐β signaling is important in pattern formation and organogenesis. During craniofacial development, TGF‐β is expressed in the craniofacial epithelium and the cranial neural crest (CNC) derived mesenchyme, suggesting tissue‐specific functions in regulating craniofacial development. To investigate the tissue‐specific requirement of TGF‐β signaling, we generated animal models with epithelim or CNC‐derived mesenchyme specific inactivation of Tgfbr2 gene and investigated the molecular mechanism of TGF‐β signaling in regulating the fate of craniofacial epithelial or the CNC mesenchyme cells, respectively. Using mice with conditional inactivation of Tgfbr2 in the epithelium, we show that TGF‐β signaling is specifically required for regulating palate fusion and skin development. At the cellular level, TGF‐β signaling is critical for mediating apoptosis of MEE cells during palatal fusion. On the other hand, conditional Tgfbr2 gene ablation in the CNC resulted in complete cleft secondary palate, calvaria agenesis, and other skull defects with complete phenotype penetrance. Significantly, disruption of the TGF‐β signaling does not adversely affect CNC migration. Cleft palate in Tgfbr2 mutant mice results from a cell proliferation defect within the CNC‐derived palatal mesenchyme. The midline epithelium of the mutant palatal shelf remains functionally competent to mediate palatal fusion once the palatal shelves are placed in close contact in vitro. Our data suggests that TGF‐β IIR plays a critical, cell autonomous role in regulating the fate of MEE and CNC cells during palatogenesis. To explore the molecular mechanism of TGF‐β signaling in regulating craniofacial development, we analyzed downstream target genes affected by altered TGF‐β signaling in either the epithelium or the CNC‐derived mesenchyme. Comprehensive analysis reveals specific TGF‐β signaling network in regulating craniofacial development. Collectively, our study demonstrates that TGF‐β signaling plays an essential role in cell fate determination of the craniofacial epithelium and the CNC‐derived mesenchyme and provides a model for the study of abnormal craniofacial development.
Inflammation is strongly associated with Plasmodium falciparum malaria and predicts erythropoietin, soluble transferrin receptor, and zinc protoporphyrin concentrations in severely anemic Zanzibari preschool children
Authors: Cusick Sarah E , Tielsch James M , Ramsan Mahdi , Jape Jape K , Sazawal Sunil , Black Robert E , Stoltzfus Rebecca J
Published: June 2021

Plasmodium falciparum malaria (Pf) contributes to severe anemia (SA) in African children, and inflammation is likely to be an important mechanism. We investigated whether Pf and inflammation predicted zinc protoporphyrin (ZPP), transferrin receptor (TfR) and erythropoietin (EPO) concentrations in a community‐based sample of 225 SA [hemoglobin (Hb) ≤ 7 g/dL] Zanzibari children (median age 13 m). Mean Hb (6.3 g/dL), ZPP (286 μmol/mol heme), serum ferritin (48 μg/L) and C‐reactive protein (CRP, 23 mg/L) suggested prevalent iron deficiency and severe inflammation. EPO was high (261 mIU/mL, normal 8–18) and appropriate for the degree of anemia. TfR was high (19 mg/L, < 8) but low for the degree of anemia, suggesting dyserythropoiesis. 76% of children had Pf‐positive blood film, which was strongly correlated with CRP (p<0.01). 17% had fever, which was related to Pf (p<0.01) and less strongly to CRP (p=0.03). Increasing CRP quartile was associated with higher EPO (p for trend = 0.02) and lower TfR (p<0.01) and ZPP (p=0.03). Increasing quartile of Pf parasitemia showed trends with EPO, TfR and ZPP that were similar in direction but less significant. In multivariate analysis with ZPP, EPO or TfR as dependent variables, Pf was not significant once CRP was added. In conclusion, inflammation strongly predicted ZPP, EPO and TfR concentrations in SA children in a manner consistent with iron sequestration and blunted bone marrow.
Independent and interactive effects of iron, lead, and parenting on children's cognitive outcomes
Authors: Hubbs‐Tait Laura , Mulugeta Afework , Bogale Alemtsehay , Kennedy Tay S. , Baker Eric R. , Stoecker Barbara J.
Published: June 2021

This study examined effects of blood lead < 10 μg/dL, iron, and parenting on children's cognition measured by Peabody Picture Vocabulary Test‐III (PPVT‐III), a measure of receptive vocabulary, and McCarthy Scales of Developmental Abilities (MCSA) verbal (reasoning, receptive and expressive vocabulary) and perceptual (pattern reproduction, conceptual grouping) scales. Participants were 112 children, mean age (SD) = 4.15 (.55) y, attending Head Start in rural Oklahoma. Venous blood samples were analyzed for lead by ICP‐MS and iron status was assessed by serum transferrin receptors. Regressions revealed significant effects of lead on MCSA perceptual scores (R2 = .04) with parental home literacy controlled (R2 = .12) and significant effects of transferrin receptors on PPVT‐III (R2 = .10) and MCSA verbal scores (R2 = .04) with parental home literacy controlled (R2 = .15 and .17, respectively). Permissive parenting (linked to poor nutrition practices) interacted with lead to predict children's MCSA perceptual scores (R2 = .04, p < .05) beyond main effects of lead and permissive parenting (R2 = .08). The interaction was due to a negative relation (β = −.551) between lead and scores for children of highly permissive parents, t(97) = −3.114, p < .01. We conclude researchers should examine parenting and nutrition in research on the impact of low levels of lead on child cognition. Funding: USDA‐CREES Number OKL0 2605.
ANALYSIS OF METABOLIC RISK FACTORS AND EATING HABIT IN OBESE MIDDLE SCHOOL STUDENTS LIVING IN KYUNG‐GI AREA
Authors: Koo Jae Ok , Jang Kyung Ja.
Published: June 2021

The purpose of this study is to identify the characteristics and metabolic risk factors of obese middle school students. The study carried out to investigate the metabolic risk factors by using the results of 500 middle school students’ (female:173, male:327) periodical anthropometric data and biological data (blood pressure, serum cholesterol, blood glucose) collected and analyzed between 2003 and 2005. The prevalence of obesity among middle school students was 11.1% for 3 years (female: 8.3%, male:13.5%). The prevalence of obesity degree (mild obesity, moderate obesity and severe obesity) was 6.3%, 5.9% and 1.2% in males and 5.1%, 2.9% and 0.3% in females respectively. There was no change in the transition of obesity for 3 years. Female students showed that total cholesterol and blood glucose had significantly increased as BMI increased. Both males and females showed significant differences in systolic blood pressure, diastolic blood pressure and blood glucose by the year. The numerical values of these tests every year therefore metabolic risk factor was considered to be elevated. The prevalence of chronic disease among obese students was 30.9%, which consists of hypertension (3.8%), hypercholesterolemia (26.3%) and hyperglycemia (0.8%). Therefore this study found metabolic risk factors among some obese middle school students and the difference between males and females.
Activation maps in the nucleus of the solitary tract (NTS) in response to internal stimuli
Authors: Schwarz Jessica , Darcel Nicolas , Rampin Olivier , Andrey Philippe , Burguet Jasmine , Fromentin Gilles , Tomé Daniel , Maurin Yves
Published: June 2021

During digestion, integration of gut information by the NTS is involved in the induction of satiation. Little is known about the encoding of peripheral information by the NTS. Similarly to other sensory encoding processes, internal sensing may rely on stimulus specific activation patterns. The aim of this study was to search for such patterns elicited in the NTS by intragastric loads of macronutrients. Male C57BL/6J mice were gavaged with isocaloric loads of either trioleate or saccharose or peptone. Animals were perfused and their brains collected and postfixed. Serial coronal brainstem sections (20 μm thick) covering the entire caudo‐rostral extent of the NTS were collected. Immunodetection of Fos protein and luxol fast blue / neutral red staining were performed on odd‐ and even‐numbered sections, respectively. Digitized brainstem images (20x) were acquired. Using Free‐D, a 3D‐reconstruction software, the contours of the brainstem and of the NTS were delineated and the Fos‐positive neurons pointed. After registration, the piling‐up of the segmentations yielded three‐dimensional models of the NTS including the activated neurons. The activation maps are compared.
Glucose lowering properties of Cordymax‐7 in DIO mice with impaired glucose metabolism
Authors: Yang Jieying , Zhao Chunsheng , Tan Ningzhi , Zhu Jia‐Shi
Published: June 2021

CordyMax (CM), a fermented product of Cordyceps sinensis, has a function of improving insulin sensitivity and glucose metabolism in animals and humans (JACM 8:309 & 8:315, 2002; CJCP 16:274, 2007). We investigated the effect of CM7 on improving glucose metabolism in DIO mice. Male ICR mice were fed high‐calorie‐diet (HCD) for 14 weeks to develop Metabolic Syndrome X (MSX) with glucose intolerance. MSX mice were randomized into 5 groups, receiving vehicle, positive drug (Metformin Hydrochloride 0.5g/kg) or CM7 (0.25, 0.5, or 1.5g/kg) for 28 days. Body weight was monitored every week. Fasting blood glucose (FBG) and oral glucose tolerance test (OGTT) were assessed at Day 28. Compared to controls, CM7 (0.5g/kg) decreased FBG by 19.3% (p=0.041) and 180‐min blood glucose by 32.6% (p=0.024) after oral dextrose bolus (1g/kg). Our data suggests that CM7 is effective in improving glucose metabolism in DIO‐MSX mice. Other animal models will be used to further assess the efficacy of CM7 in glucose metabolism before clinical tests in humans.
Signal transduction links intramyocellular triglycerides to muscle insulin resistance in obesity
Authors: Guo ZengKui
Published: June 2021

In obesity, both the content and turnover of intramyocellular triglycerides (imcTG) in skeletal muscle are increased and the abnormality is associated with muscle insulin resistance (MIR). Diacylglycerol (DAG) and ceramide (Cer) in myocytes are also elevated; they act on PKC system to impair insulin sensitivity. To test the hypothesis that imcTG increases DAG and Cer fluxes thereby causing MIR, 14C‐triolein and 3H‐phosphatidylcholine were incubated in homogenate of gastrocnemius muscle from lean (n=4) and obese (n=4) rats. Plasma membrane and cytosol were prepared from incubated homogenates, the lipids extracted, DAG and Cer isolated by TLC and their 14C‐radioactivity was used to represent their flux from imcTG. DAG flux to plasma membrane in obese rats was greater (10.1±2.1 vs. 1.6±0.3 nmol/g•min, P<0.01), so was DAG flux to cytosol (55±6.1 vs. 3.9±0.7 nmol/g•min, P<0.01). In contrast, DAG flux to plasma membrane from phospholipids (PL) was lower in obese rats (0.12±0.02 vs. 0.28±0.1 nmol/g•min, P<0.05), consistent with reduced PL content (Horm. Metab. Res. 37:773, 2005). Cer flux to plasma membrane in obese rats was also higher (0.23±0.01 vs. 0.17 0.02 nmol/g•min, P<0.05) whereas the flux from PL was lower (0.02±0.005 vs. 0.06±0.02 g·min, P<0.05). The novel finding indicated that imcTG is a significant and preferred source of DAG and Cer in myocytes of obese rats. Therefore, imcTG contributes to MIR via PKC signaling pathways, explaining the imcTG‐MIR correlation.
Bone Mineral Density in Hutterite Children
Authors: Wey Christine , Binkley Teresa , Beare Tianna , Arneson Lacey , Specker Bonny
Published: June 2021

We previously found that Hutterite women and men have greater areal bone mineral density (aBMD) than non‐Hutterites. It is unknown whether this higher aBMD occurs at younger ages. We examined Hutterite children to test the hypotheses that aBMD in younger (<15y) children would be similar to reference data (Z score = 0); but greater than reference data (Z score>0) in older children after they enter the adult workforce at age 15. We measured hip and spine aBMD in 325 children aged 5 to 20 years: 186 children <15y and 139 children >=15 years. Older Hutterite females and males had higher % time in moderate+vigorous activity (21±10%, 29±11% respectively [mean±SD]), than younger females and males (15±10%, 18±10%). Younger females and males had hip Z‐scores (0.30±1.0, 0.44±1.0) greater than 0 (p = 0.002, p<0.001 respectively) indicating higher aBMD among young children compared to reference data. Younger males had spine Z‐score (−0.27±1.2) less than 0 (p = 0.04). None of the Z‐scores for the older age group differed from 0. Contrary to our hypothesis, younger Hutterite children had greater hip aBMD Z scores than the reference data; whereas older children did not. We speculate, based on previous data in adults that younger menarcheal age in this population may lead to greater hip Z scores among the younger females. The reason for low spine aBMD in young Hutterite males is being investigated.
Associations of maternal folate intake with antepartum and postpartum serum folate levels
Authors: Kim Hyesook , Lee Ah Young , Chang Namsoo
Published: June 2021

This study was carried out to evaluate the effect of maternal folate intake on the maternal serum folate level. Pregnant women of 18–20 weeks of gestation were recruited from prenatal clinic in Seoul, Cheonan and Ulsan. In 429 pregnant women, dietary intake during pregnancy was assessed by a 24‐hour recall method. Serum folate level was analyzed by the radioimmunoassay(RIA). The daily mean energy and protein intakes were 1804.9 kcal (90.9% of EER), 68.1g (125.8% of EAR, 97.0% of RI), respectively. The average daily folate intake was 238.2 μg DFE (45.95% of EAR, 39.82% of RI) for the pregnant women. The maternal antepartum(18–20 wk) and postpartum(at delivery) serum folate concentration were 10.4 ± 7.9 ng/ml, 13.5 ± 9.3 ng/ml. The maternal antepartum serum folate level was significantly lower than that of the postpartum serum(p<0.001). Pregnant women whose intake of folate was above the median value showed a greater increase in postpartum serum folate level than those with intakes below the median(p<0.05). Maternal intake of folate was positively correlated with postpartum serum folate level (p<0.05). These results suggest that the maternal folate intake may affect the postpartum folate level.

This work was supported by the Ministry of Environment and the Brain Korea 21 project in 2007
Integration of Embryology and Histology
Authors: Pawlina Wojciech , Ross Michael H. , Carmichael Stephen
Published: June 2021

Developmental anatomy can be used to organize the teaching of histology. Since certain tissues are derived from predictable embryonic layers or structures, the embryological organization can be used to organize tissues in a meaningful and useful manner. We offer several examples of this relationship to help students organize tissues by their developmental origins. First, all functional neurons originate from ectoderm, but the fate of cells coming from the neural tube differs from those that come from the neural crests. All neurons that have their final location within the central nervous system (CNS) are derived from the neural tube, whereas neurons whose nerve cell bodies are outside the CNS come from the neural crests. Second, almost all muscle tissue, which comprises about 40% of the human body mass, comes from the mesoderm. The exceptions are the smooth muscle cells within the eye, which are derived from the neural crest. Almost all connective tissues, with the exception of the notochord, are derived from the mesoderm, or more specifically from mesenchymal cells. Melanocytes (also satellite cells and Schwann cells) are derived from neural crest. Epithelial cells that contribute to the epidermis are derived from ectoderm, and almost all other epithelial tissues are derived from endoderm. Exceptions are endothelial cells (including endocardium) that come from mesoderm. This outlines the close relationship between developmental anatomy and histology that helps to categorize tissues by their embryologic origin.
Dietary chlorella supplementation improves oxidative stress induced by cadmium in SD rats
Authors: Hong Soyoung , Son Young Ae , Kim Mi Kyung
Published: June 2021

This study was designed to investigate the effect of chlorella supplementation on oxidative stress induced by dietary cadmium (Cd) in Sprague‐Dawley (SD) rats. Sixty male SD rats (14 week‐old) were fed diets containing 0%, 3% or 5% chlorella, and 0 ppm (−Cd) or 160 ppm (+Cd) Cd for 10 weeks. Serum and hepatic superoxide radical generation, erythrocyte and hepatic antioxidative enzyme activities (SOD, GPx, and catalase), and plasma MDA and renal 8‐hydroxy‐2′‐deoxyguanosine levels were measured. Serum superoxide radical generations of +Cd groups were significantly higher than −Cd groups, and 3% chlorella +Cd group was significantly lower than 0% chlorella +Cd group. Hepatic superoxide radical level in 5% chlorella −Cd group was significantly lower than 0% chlorella−Cd group. But, erythrocyte and hepatic SOD acitivities showed no difference among all the groups. Plasma MDA levels of 0% chlorella +Cd group were significantly higher than 0% chlorella −Cd group, and reduced in 3% chlorella +Cd group compared with 0% chlorella +Cd group. There findings suggest that chlorella supplementation showed antioxidative capacity against oxidative stress induced by dietary Cd. *This work was supported by the grants from Daesang.Co.Ltd. and the second stage of Brain Korea 21 project in 2007.
Antioxidative Potential of Soybean Varieties with Different Levels of Isoflavones and Anthocyanins
Authors: Kim Jong‐Sang , LIM Jisun , Seo Ji Yeon , Lee InAE , Hwang Young Hyun
Published: June 2021

There is ample evidence that soybean has antioxidative activity and protect tissues from oxidative stress‐induced injury. Although isoflavones present in soy are believed to be major components responsible for the antioxidative activity, our recent study showed that anthocyanins present in black soybean had strong antioxidative potential. We, therefore, were interested in investigating whether black soybean possesses stronger antioxidant potential than yellow counterpart. Seven groups of mice were employed in the present study, each comprising of 10 mice. Group 1, 2, 3 received water injection (control), Fe‐NTA (8 mg/kg bw), Fe‐NTA + alpha‐tocopherol (2000 U), respectively, while group 4, 5, 6, 7 were fed diets containing 10% (w/w) normal yellow soy, normal black soy, high‐isoflavone soy, black high‐isoflavone soy, respectively, from a week before Fe‐NTA injection to one week after the injection. Biomarkers for oxidative stress including superoxide dismutase, catalase, glutathione peroxidase, phosphoglycerate kinase 1, TBARS, FRAP were compared among tissues collected from experimental groups. Overall, high‐isoflavone and black high isoflavone varieties of soy appear to effectively protect the animals from Fe‐NTA‐induced oxidative stress.
Efficacy of Juice Powder Concentrates on Redox State in Heavy Smokers
Authors: Bamonti Fabrizia , Sedda Valentina , Dellanoce Cinzia , Gregori Dario , Schiraldi Gianfranco , Novembrino Cristina , Guinea Marie , Accinni Roberto
Published: June 2021

Smoking can contribute to human pathogenesis through the production of reactive oxygen species, and also by weakening the antioxidant system. To study the efficacy of supplementation with dehydrated concentrated of fruit and vegetable juice powder (FVJ) and the same plus berry (FVJ+), on the oxidative balance, 61 healthy heavy smokers (35 males, age 40–60y; 20 cigarettes/day or more for at least 10 y) were randomized into 3 groups: A (placebo, n=19), B (FVJ, n=21), C (FVJ+, n=21). They were supplemented for 12 weeks and underwent blood sampling at baseline (T0), and at end of supplementation (T1), for plasma and blood thiols HPLC analysis. No differences between groups’ aminothiol concentrations were found at T0. Total homocysteine (Hcy) decreased significantly in all compartments both in group A and B (plasma: p<0.001 for A and B; blood: p=0.027 A vs C, and p=0.018 B vs C), as well as the reduced Hcy (plasma: p=0.005 A vs C; blood: p=0.005 A vs C, and p<0.001 B vs C). Only group B showed a significant decrease of total blood glutathione (GSH) compared to group C (p=0.027). Supplementation with FVJ effectively improved Hcy metabolism, and concomitantly folate levels. Also FVJ+ supplementation was effective in improving GSH cycle.
Cigarette smoke induces cPLA2 expression via NADPH oxidase, MAPKs, AP‐1 and NF‐κB in human airway smooth muscle cells
Authors: Cheng Shin‐Ei , Jou Mei‐Jie , Kou Yu‐Ru , Yang Chuen‐Mao
Published: June 2021

Cigarette smoking (CS) causes a variety of airway inflammatory diseases, including chronic obstructive pulmonary disease and asthma. After allergens stimulation, increased of cPLA2 expression may play a critical role in airway inflammation. However, the mechanisms underlying CS‐induced cPLA2 expression in airway smooth muscle cells (ASMCs) are still unknown. In this study, CS‐induced NADPH oxidase activation and superoxide production, led to increase cPLA2 expression in human ASMCs. The mechanism of cPLA2 expression is dependent on MAPKs phosphorylation, NF‐κB and AP‐1 activation. Pretreatment with the inhibitors of NADPH oxidase (DPI, apocyanin), N‐acetylcysteine, MEK1/2 (UO126), p38 MAPK (SB202190), and JNK (SP600125) suppressed CS‐induced cPLA2 expression, revealed by RT‐PCR and Western blotting analyses. Transfection with siRNAs of JNK1, p42 and p38 also reduced cPLA2 expression. Induction of cPLA2 by CS was inhibited by a NF‐κB inhibitor (helenalin) and a selective inhibitor of AP‐1 (curcumin). Moreover, promoter assays revealed that increases of cPLA2, NF‐κB and AP‐1 luciferase activities in response to CS were attenuated by these inhibitors. These results suggested that in ASMCs, activation of NADPH oxidase, MAPKs pathway, NF‐κB, and AP‐1 is essential for CS‐induced cPLA2 expression.
Prevalence of human papillomavirus (HPV) infection in a cohort of young women in Delhi, India
Authors: Datta Palika , Patro Raj Kumar , Bhatla Neerja , Singh Neeta
Published: June 2021

Introduction: HPV is strongly associated with cervical cancer and is highly prevalent in sexually active women. In Indian women, it affects nearly 132,000 and kills 76,000 annually. Geographical variations in the epidemiological aspects of HPV such as incidence, prevalence, age and type distribution are seen and knowledge of these variables in a region‐specific context is necessary for designing effective interventional strategies.

Objective: To determine the prevalence of HPV infection and associated genotypes in cervical samples in Northern India.

Methodology: 800 exfoliated cervical swab samples were collected from unscreened married women aged 16–24 years. HPV testing was done by Hybrid Capture assay 2 and PCR using PGMY L1 consensus primers. HPV genotyping was done for positive samples by Reverse Line blot array.

Results: HPV DNA was detected in 69 (8.6%) samples. In HPV positive samples, type 16 was seen in 36.2% and HPV 51 and 59, 13% each. A single HPV type was found in 40 (58%) cases and mixed infection seen in 29 (42%).

Conclusions: Multiple high‐risk HPV infections were most prevalent in young women and HPV‐16 was the dominant type. Etiologic role and persistence are being studied by doing 6 monthly follow up. HPV incidence, prevalence and type can give an indication of the natural history of HPV infection in the younger age group. This study was supported by the Department of Biotechnology, Government of India.
Morphometric Evaluation of Colon Crypts in Experimental DMH Administration
Authors: Cabrita Antonio Silverio , Farinha Rpdrigo , Rafael Ana Isabel , Matos‐Beja Manuel Correia
Published: June 2021

In this study we evaluate changes in the bowel of animals submitted to the administrations of DMH in a colon cancer induction protocol, by studying how this carcinogen affects the morphology of the crypts.

We used 60 Wistar male rats, four weeks old separated in three groups, as follows: group I, kept with no manipulation (control group); group II submitted to the administration of 15 mg/Kg DMH twice a week, for eight weeks, after which they were sacrificed; group III submitted to the same conditions of group II, sacrificed after 4 weeks after the last administration of DMH. We calculated the colon CWHR (Crypt width – height ratio) for three different sections ‐ proximal, medial and distal.

For the proximal region, CWHR for a 95% confidence interval varies between 0.172 and 0.225 for group I, between 0.178 and 0.204 for group II and 0.132 and 0.156 for group III.

For the medial region, CWHR for a 95% confidence interval varies between 0.133 and 0.177 for group I, between 0.146 and 0.191 for group II and between 0.108 and 0.140 for group III.

For the distal region, CWHR for a 95% confidence interval varies between 0.120 and 0.288 for group I, between 0.313 and 0.401 for group II ans bwtween 0.137 and 0.177 for group III.

According to these results, the tested carcinogen affects the colon crypts morphology early after the administration.
+ACA BRCA1 promoter polymorphism confers increased risk of breast cancer among African‐American women
Authors: White Kristen K. , Basta Patricia V. , Booker Jessica K. , Millikan Robert C. , Coleman William B.
Published: June 2021

We previously identified an ACA insertion polymorphism (+ACA) in the BRCA1 promoter that creates a consensus binding site (AACAACAC) for the FAC1 transcriptional repressor (located 600 bp upstream of the exon 1a transcriptional start site). Expression of FAC1 in individuals that carry the +ACA polymorphism may result in loss of BRCA1 expression, contributing to breast cancer susceptibility. Genotyping of 589 women from the Carolina Breast Cancer Study (308 cases, 281 controls) yielded higher +ACA allele frequencies among African‐American (AA) cases (27%) compared to AA controls (17%, P = 0.0005), but there was no significant difference among Caucasian (CC) cases and controls (34% versus 37%, P = 0.50). Genotype distributions among AA and CC cases and controls showed no significant departures from Hardy Weinberg equilibrium. Age‐adjusted odds ratios (ORs) for breast cancer among AA women were 4.6 (95% CI 1.3–17.0) for +ACA/+ACA and 1.6 (95% CI 1.0–2.7) for wt/+ACA compared to wt/wt homozygotes. Among CC women, the relevant ORs were 0.7 (95% CI 0.3–1.5) for +ACA/+ACA and 1.1 (95% CI 0.7–1.8) for wt/+ACA. These results suggest a statistically significant association between the +ACA BRCA1 promoter polymorphism and breast cancer susceptibility among AA women.

Support: NIH CA78343, CA58223, CA16086; Komen Foundation BCTR0100575, Friends For an Earlier Breast Cancer Test.
Hepatitis C treatment based on differential neural estimator
Authors: CABRERA AGUSTIN IGNACIO , GOMEZ YOLANDA , BAUTISTA MARIA ESTHER , RAMIREZ‐SOTELO MARIA GUADALUPE
Published: June 2021

Hepatitis C is suffered approximately by the world population's three percent and it has become an important social problem that has shown to be in continuous growing, for each case the treatment is different so a general procedure is not applicable to this sickness. For that reason develop new ways to treatment are needed. This report show the application of the Hepatitis C Virus mathematical model and the differential neural networks (DNN) to develop a system estimator in order to rebuild the states variables of this sickness and the design a control function based on Hamilton Jacobi Technique (HJT), a technique used in control systems. The estimator was trained just one time with the information given of a sick patient without previous treatment. This control is a tool that gives us an approximation of the dosage of Interferon Alpha 2β (IFN‐2β) to be prescribed to a patient in order to maintain the viral load in low levels. This algorithm was tested by numerical simulations and the good performance shows us the possibility to develop Hepatitis C treatments for medications effective dose.
Antibody microarray longevity study: How long‐term storage affects the dynamic range of printed slides
Authors: Bickel Daniela , Warner Roscoe L. , Johnson Kent J.
Published: June 2021

Following development of an antibody microarray we undertook a six month study of a single batch of chips to determine the loss of sensitivity over time. Picogram amounts of capture antibodies are spotted onto epoxy‐silane slides using a Piezorray printer. A single printing, of eleven slides containing 30 anti‐rat antibodies was stored at 4°C in a desiccator until used. A freshly printed slide was analyzed to assure accuracy of print. During the six month test‐period slides were arrayed by first hybridizing with protein standards in separate wells of a pro‐plate system and bound proteins detected using biotin‐conjugated secondary antibodies. Streptavidin conjugated Alexafluor is used to generate the fluorescent signal. A single slide was analyzed with protein standards every two weeks, starting 3 weeks post‐printing and ending at the 6 month time‐point. A single slide was stored at room temperature in a desiccator and tested 3 months post‐printing. We found less than a 2% change in the detection threshold relative to the dynamic range of the standard curves for the slides stored at 4°C. While under the same conditions the slide stored at room temperature exhibited a 15% degradation at 3 months. These findings demonstrate that antibody microarrays can be used for extended periods of time post‐printing, allowing for a single batch of chips to be used for large ongoing studies. Work supported by a grant from Pfizer of Ann Arbor.