bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lytic granule‐mediated, noncytolytic CD8+ T cell inhibition of HSV‐1 reactivation from neuronal latency
Authors: Knickelbein Jared E. , Hendricks Robert L. 
Published: June 2021

The purpose of our studies is to identify and characterize the mechanisms employed by HSV‐specific CD8+ T cells (HSV‐CD8) to block HSV‐1 reactivation from latency in sensory ganglia, potentially limiting recurrent herpetic disease. HSV‐CD8 can secrete IFN‐γ to inhibit reactivation in some, but not all neurons, suggesting involvement of additional effector mechanisms. We now show that the lytic granule components perforin (Pfn) and granzyme B (GrB) are necessary for optimal protection from HSV‐1 reactivation in sensory neurons both in vivo and in ex vivo trigeminal ganglia (TG) cultures. We further establish that HSV‐CD8 exhibit vectorial release of lytic granules into immune synapses formed with both susceptible fibroblasts and latently infected neurons in ex vivo TG cultures. Importantly, while lytic granule release activates the caspase system of fibroblasts leading to morphologic signs of apoptosis, similar lytic granule release does not induce caspase activation or morphologic signs of apoptosis in neurons. Together our findings establish that directed lytic granule release by HSV‐CD8 is required to block neuronal HSV‐1 reactivation, that lytic granules block HSV‐1 reactivation in neurons without activating caspases or inducing apoptosis, and that GrB is one (but apparently not the only) lytic granule component required to block HSV‐1 reactivation.Supported by NIH, Eye & Ear Foundation, & RPB.
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.
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.
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
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.
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
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
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.
Nucleus accumbens dopamine and it's role in positively reinforcing aggression
Authors: Couppis Maria Helena , Kennedy Craig H. 
Published: June 2021

Rationale: We studied whether mesocorticolimbic dopamine (DA) in the nucleus accumbens (NAc) mediates the rewarding properties of aggression.Objective: To address this issue, we studied the effect of administrating specific DA receptor antagonists into the NAc on aggression and general movement.Methods: A residentintruder model of aggression was used with male mice. An operant conditioning paradigm was used to establish a baseline rate of nose pokes to contingently access aggression as a positively reinforcing stimulus. We analyzed whether microinfusions into the NAc of SCH‐23390, a D1 like DA receptor antagonist, or sulpiride, a D2 like DA receptor antagonist, altered aggression and/or movement. Open‐field tests were used to further assess the effects of SCH‐23390 and sulpiride on movement.Results: Infusing either SCH‐23390 or sulpiride decreased responding for contingent aggression in a dosedependent fashion without disrupting general movement. Infusing SCH‐23390 or sulpiride did not disrupt movement in the open‐field tests at dosages reducing operant responding in the previous aggression tests.Conclusions: Our results suggest that both D1‐like and D2‐like DA receptors in the ventral striatum may mediate the rewarding properties of aggression as a positively reinforcing stimulus.
Smooth muscle specific knockdown of L‐type calcium channel by exogenous microRNA (miRNA)
Authors: Pang Li , Stimers Joseph , Rhee Sung , Palade Philip 
Published: June 2021

In spontaneously hypertensive rats, vascular L‐type calcium channel pore forming α1 subunit (Cav1.2) protein expression, L‐type calcium current (CaL) and vascular tone are all increased, due to increased expression of the ubiquitously expressed isoform containing an exon 1b encoded N‐terminus. The objective of this study is to develop shRNA or miRNA constructs against the non‐cardiac form of Cav1.2 (exon1b isoform) to reverse the upregulation in a tissue‐specific manner. Cav1.2 exon 1b‐specific shRNA, miRNA, and appropriate controls were constructed and tested for silencing effects. In HEK cells, rat Cav1.2 expression vector (rCav1.2) was cotransfected with different constructs, CMV promoter‐driven miRNA (Cav1.2) had the strongest effect and decreased rCav1.2 protein ∼72%. More interestingly, miRNA(Cav1.2) driven by the vascular smooth muscle‐specific promoter SM22α reduced endogenous Cav1.2 expression (western blot) ∼51% and decreased CaL (barium) current (patch clamp) ∼46% in transfected A7r5 cells. Moreover, the SM22‐GFP‐miRNA(Cav1.2) had no effect on cotransfected rCav1.2 expression in HEK cells and showed little or no GFP expression in PC12 or HL1 cells.Conclusion:SM22–GFP‐miRNA(Cav1.2) can effectively and smooth muscle‐specifically reduce Cav1.2 expression. Coupled to adeno‐associated virus, this may provide a novel long‐term therapy for hypertension. Supported by NIH HL63903.
Modification of a calcein‐based assay for monitoring proteolytic activity in tissue suffusate
Authors: Mikel Kyle , Bingaman Susan , Huxley Virginia H. 
Published: June 2021

This investigation was aimed at developing an assay to characterize protein degradation in effluent collected from tissues on which microvascular permeability studies were being conducted (Bingaman S., FASEBJ: 20, 2006). In particular, we were measuring protein clearance from venules and assaying total protein in the effluent. The assumption we wanted to test was whether the protein levels reflected intact material, whether proteolysis was occurring, and whether the treatments which altered protein clearance reflected changes in proeolytic activity. The approach we used was based on the quenching of calcein (Ex 450 nm, Em 530 nm) by Cu(II) and the ability of proteolytic products (amino acids and peptides) to coordinate with the Cu(II), removing it from the calcein, allowing it to fluoresce (Dean, K., Biorganic & Medicinal Chem Letters, 2003). After optimization of assay conditions, samples collected from representative leak index experiments (Bingaman S., FASEBJ: 20, 2006) were tested. The results showed a clear difference between samples with respect to the distribution of proteins and small peptides under both control and test conditions. Therefore, it can be concluded that this assay can be used to screen for proteolytic activity in the microvasculature.Supported by NIH HLO78816, NASA NN04GA50G, AHA 0615515Z (RS), NIHCO6RRO17353 and Missouri Academy
Transforming Growth Factor‐β1 Inhibits CFTR‐dependent cAMP‐stimulated Alveolar Epithelial Fluid Transport: Critical Role of Interleukin‐8
Authors: Roux Jeremie , Ganter Michael T , Goolaerts Arnaud , Finkbeiner Walter , Matthay Michael A , Pittet Jean‐Francois 
Published: June 2021

β‐adrenergic agonist‐dependent stimulation of the lung fluid clearance is an important mechanism that protects the lung from alveolar flooding. In this study we hypothesized that critical mediators of acute lung injury (ALI), such as transforming growth factor‐β1 (TGF‐β1) and interleukin‐8 (IL‐8), could directly antagonize the epithelial response to β‐adrenergic agonists. Short circuit current experiments revealed that TGF‐β1 inhibits CFTR‐specific β‐agonist‐stimulated vectorial Cl‐transport across the apical membrane of primary rat and human alveolar epithelial type II (ATII). TGF‐β1 also significantly decreased β‐agonist‐stimulated cAMP production resulting in the inhibition of the CFTR promoter activity and gene expression in ATII cells. We found that this TGF‐β1‐dependent inhibition required IL‐8 and was mediated by desensitization the β‐adrenergic receptor through both TGF‐β1 and IL‐8 signaling pathways. Consistent with the in vitro results, we showed that TGF‐β1 requires IL‐8 to inhibit the β‐agonist‐stimulated fluid transport across the distal airspace epithelium in vivo in rats. In summary, the results show for the first time that TGF‐β1 and IL‐8 inhibit the stimulated vectorial chloride transport across the distal lung epithelium, thus impairing the β‐adrenergic‐dependent protective mechanism against alveolar flooding.NIH P50HL074005 (JFP), ALA Award and T32 GM008440 (JR)