Analysis and Development of Antigen-specific T Cells Derived from Peripheral Blood Mononuclear Cells of Healthy Donors.

BACKGROUND/AIM: Adoptive cell therapy using antigen-specific T cells is a promising treatment modality for cancer patients. Various methods to isolate specific T cells and identify corresponding T cell receptor (TCR) sequences are known. This study aimed to identify antigen-specific TCR from T cells isolated using carboxyfluorescein succinimidyl ester (CFSE), which marks proliferating activated T cells. MATERIALS AND METHODS: CFSE stained healthy donor peripheral blood mononuclear cells (PBMCs) were treated with cytomegalovirus (CMV) or Epstein-Barr virus (EBV) peptides for seven days. Then, proliferating T cells with decreased CFSE staining were isolated and single cell VDJ sequencing was performed on isolated T cells to identify antigen-specific TCRs. RESULTS: As antigen-specific TCR candidates, ten TCR clones were selected for the CMV antigen and five for the EBV antigen. The reactivity of ten CMV TCR-transduced T cells and one EBV TCR-transduced T cell toward T2 cells pulsed with CMV or EBV peptide was confirmed via NFAT-luciferase, IFN-γ ELISA, and cytotoxicity assays. CONCLUSION: Identification of antigen-specific TCRs with CFSE staining is a valid method for the development of effective immunotherapy. The identified CMV- or EBV-specific TCRs can be used for adoptive cell therapy to treat cancer.

Effects of Grape Seed Extract Supplementation on Endothelial Function and Endurance Performance in Basketball Players.

While dietary polyphenols supplements can improve endothelial function and blood flow to exercise, the effects of chronic supplementation with grape seed extract (GSE) containing a high dose of polyphenols on endurance performance are not known. Accordingly, in 12 elite athletes, we compared the effects of both GSE and placebo (PL) on submaximal VO2, time to exhaustion performance, and endothelial function during progressive cycling exercise for 14 days. Endothelial function was evaluated from the brachial artery via flow-mediated dilation (FMD). Compared to PL, GSE decreased submaximal VO2 at 80% and 120% of VO2peak and increased the time to exhaustion (p < 0.05). GSE also resulted in FMD-induced increase in brachial artery diameter (14.4 ± 5.2% vs. 17.6 ± 4.5%, p = 0.035). We demonstrated that chronic supplementation with GSE improved endurance performance and these effects may partially be due to vasodilation in active skeletal muscle mediated by enhanced endothelial function. Thus, our results suggest that GSE appears to be an ergogenic nutraceutical that can improve exercise performance in elite athletes.

Effects of Somatic, Depression Symptoms, and Sedentary Time on Sleep Quality in Middle-Aged Women with Risk Factors for Cardiovascular Disease.

Cardiovascular disease (CVD) is the second leading cause of death among Korean women, and its incidence is dramatically elevated in middle-aged women. This study aimed to identify the predictors of sleep quality, a CVD risk factor, in middle-aged women with CVD risk factors to provide foundational data for developing intervention strategies for the prevention of CVD. The subjects, 203 middle-aged women (40-65 years old) with one or more CVD risk factors were selected through convenience sampling and included in this descriptive correlational study. The effects of somatic symptoms, depression symptoms, and sedentary time on sleep quality were examined. CVD-related characteristics were analyzed using descriptive statistics, whereas the mean values of the independent variables were analyzed using t-tests and analysis of variance. Predictors of sleep quality were analyzed using multiple regression analysis. The results showed that sleep quality increased with decreasing somatic symptoms (ß = -0.36, p < 0.001), depression symptom score (ß = -0.17, p = 0.023), and daily sedentary time (ß = -0.13, p = 0.041), and the regression model was significant (F = 19.80, p < 0.001). Somatic symptoms are the most potent predictors of sleep quality in middle-aged women. Thus, intervention strategies that improve somatic symptoms are crucial for the enhancement of sleep quality, which deteriorates with advancing age.

Investigation of Pathogenic Escherichia coli Contamination of Vegetables Distributed in a Korean Agricultural Wholesale Market.

The purpose of this study was to investigate the monthly contamination rate of pathogenic Escherichia coli, a major cause of food poisoning, in vegetables sold in agricultural wholesale markets, which distribute vegetables from all over the country, in the Incheon Metropolitan City area, South Korea, and to identify a source of the pathogen. In total, 1739 vegetables of 80 types, along with 109 soil, 67 manure, and 33 livestock feces samples, were tested for pathogenic E. coli using polymerase chain reaction, from September 2016 through August 2017. The average annual prevalence rate of vegetables was 5.8%, and the prevalence rate was above 5% from June through October. The highest prevalence rate (15.7%) was recorded in July. Water dropwort showed the highest prevalence rate (28.6%) among the vegetables examined. Pathogenic E. coli was detected in >20 types of the vegetables that were to be consumed without cooking. Among these, the prevalence rates of ponytail radish (n = 21), crown daisy (n = 86), young radish (n = 68), romaine lettuce (n = 133), perilla leaf (n = 103), Korean leek (n = 43), young Chinese cabbage (n = 68), and Chinese cabbage (n = 30) were 9.5%, 8.1%, 7.4%, 6.8%, 4.9%, 4.7%, 4.4%, and 3.3%, respectively. Among the vegetables cooked before consumption, prevalence rates were 28.6%, 27.3%, and 25.0% in wormwood, sweet potato stalk, and edible mountain vegetables (Saussurea sp., etc.), respectively. In soil, manure, and livestock feces, 36.7%, 26.9%, and 90.6% prevalence rates were confirmed, respectively. This study confirmed the pathogenic E. coli contamination of vegetables to be consumed without cooking. Therefore, to produce agricultural products that do not induce food poisoning and are safe for consumption, it is important to develop a process for killing the pathogenic microorganisms and set up a sanitary environment for effectively managing compost. In addition, it is necessary to establish surveillance systems to monitor the production chain.

A putative exosporium lipoprotein GBAA0190 of Bacillus anthracis as a potential anthrax vaccine candidate.

BACKGROUND: Bacillus ancthracis causes cutaneous, pulmonary, or gastrointestinal forms of anthrax. B. anthracis is a pathogenic bacterium that is potentially to be used in bioterrorism because it can be produced in the form of spores. Currently, protective antigen (PA)-based vaccines are being used for the prevention of anthrax, but it is necessary to develop more safe and effective vaccines due to their prolonged immunization schedules and adverse reactions. METHODS: We selected the lipoprotein GBAA0190, a potent inducer of host immune response, present in anthrax spores as a novel potential vaccine candidate. Then, we evaluated its immune-stimulating activity in the bone marrow-derived macrophages (BMDMs) using enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Protective efficacy of GBAA0190 was evaluated in the guinea pig (GP) model. RESULTS: The recombinant GBAA0190 (r0190) protein induced the expression of various inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-1α (MIP-1α) in the BMDMs. These immune responses were mediated through toll-like receptor 1/2 via activation of mitogen-activated protein (MAP) kinase and Nuclear factor-κB (NF-κB) pathways. We demonstrated that not only immunization of r0190 alone, but also combined immunization with r0190 and recombinant PA showed significant protective efficacy against B. anthracis spore challenges in the GP model. CONCLUSIONS: Our results suggest that r0190 may be a potential target for anthrax vaccine.

Predictive Factors on the Incidence of Heart Failure in Patients with Ischemic Heart Disease: Using a 10-Year Population-Based Korea National Health Insurance Cohort Data.

Early risk stratification and preventative strategies are required in patients with ischemic heart disease (IHD) to prevent heart failure (HF). We aimed to investigate the rate of progression to HF and to investigate the factors predicting the development of HF in a population with IHD for 10 years. A descriptive study was conducted using Korea National Health Insurance Service-National Sample Cohort (NHI-NSC) data (2005-2015). Among the patients diagnosed with IHD for the first time in 2005-2006, 2271 men and 2037 women who responded to the health check-up survey were finally selected. Cox Proportional Hazard regression analyses and the Kaplan-Meier survival analysis were used. HF incidence rates were 5.1% in men and 8.0% in women. The mean duration of transition to HF was 4.85 ± 2.73 years in men and 4.73 ± 2.73 years in women. The non-incidence rate of HF was higher in men than in women (Log-rank test, p = 0.0003). Bivariate analyses showed that older age, prevalence of hypertension and diabetes, less alcohol, and lower physical exercise were associated with the incidence of HF in both men and women. Multivariate analyses found that HF incidence in aged subjects ≥70 years was 1.46 times higher in men and 1.44 times higher in women compared to those in their 30 s (p < 0.001). Prevalence of hypertension reduced the incidence of HF by 0.78 and 0.87 for men and women, respectively. The prevalence of diabetes increased 1.23 times only in men. These findings suggest that special attention such as periodic counseling and education is needed to prevent progression to HF in elderly and diabetic patients during follow-up after IHD.

Monitoring Culicine Mosquitoes (Diptera: Culicidae) as a Vector of Flavivirus in Incheon Metropolitan City and Hwaseong-Si, Gyeonggi-Do, Korea, during 2019.

The flaviviruses are small single-stranded RNA viruses that are typically transmitted by mosquitoes or tick vectors and are etiological agents of acute zoonotic infections. The viruses are found around the world and account for significant cases of human diseases. We investigated population of culicine mosquitoes in central region of Korean Peninsula, Incheon Metropolitan City and Hwaseong-si. Aedes vexans nipponii was the most frequently collected mosquitoes (56.5%), followed by Ochlerotatus dorsalis (23.6%), Anopheles spp. (10.9%), and Culex pipiens complex (5.9%). In rural regions of Hwaseong, Aedes vexans nipponii was the highest population (62.9%), followed by Ochlerotatus dorsalis (23.9%) and Anopheles spp. (12.0%). In another rural region of Incheon (habitat of migratory birds), Culex pipiens complex was the highest population (31.4%), followed by Ochlerotatus dorsalis (30.5%), and Aedes vexans vexans (27.5%). Culex pipiens complex was the predominant species in the urban region (84.7%). Culicine mosquitoes were identified at the species level, pooled up to 30 mosquitoes each, and tested for flaviviral RNA using the SYBR Green-based RT-PCR and confirmed by cDNA sequencing. Three of the assayed 2,683 pools (989 pools without Anopheles spp.) were positive for Culex flaviviruses, an insect-specific virus, from Culex pipiens pallens collected at the habitats for migratory birds in Incheon. The maximum likelihood estimation (the estimated number) for Culex pipiens pallens positive for Culex flavivirus was 25. Although viruses responsible for mosquito-borne diseases were not identified, we encourage intensified monitoring and long-term surveillance of both vector and viruses in the interest of global public health.

Augmented Hemodynamic Responses in Obese Young Men during Dynamic Exercise: Role of the Muscle Metaboreflex.

Studies found that cardiovascular responses to exercise are enhanced in individuals with obesity and are associated with a greater cardiac output (CO) response compared to normal weight controls. However, the mechanisms underlying these altered responses during dynamic exercise are not clear. We investigated whether the cardiovascular responses mediated by the muscle metaboreflex (MMR) activation are augmented in obese men during both static and dynamic exercise. Twenty males (10 obese (OG) and 10 non-obese (NOG)) were studied. Changes in CO, mean arterial pressure (MAP), and total vascular conductance (TVC) were compared between the two groups during dynamic handgrip exercise (DHE), post-exercise muscular ischemia (PEMI), and dynamic exercise corresponding to 40%, 60% and 80% workloads. Subjects completed 2 min of DHE at 30% of MVC, followed by 2 min of PEMI. MAP, CO, and TVC responses to DHE and dynamic exercise were significantly higher in OG, whereas there were no differences during PEMI. Increases in CO and MAP during mild to heavy dynamic exercise were seen in both groups, but the changes in these variables were greater in the OG. There were no significant differences in TVC between the two groups. Compared to NOG, the augmented blood pressure response to DHE and dynamic exercise in OG was associated with a greater increase in CO. Thus, the augmented CO and MAP responses were not associated with the activation of the MMR. Consequently, additional factors specific to obesity, such as the mechanoreflex, may have been involved.

Pure Laparoscopic Versus Open Left Hepatectomy Including the Middle Hepatic Vein for Living Donor Liver Transplantation.

Pure laparoscopic donor hepatectomy (PLDH) has become increasingly accepted in the era of minimally invasive surgeries. However, the outcomes of pure laparoscopic donor left hepatectomy (PLDLH) are relatively less known than for left lateral sectionectomy or right hepatectomy. This study aimed to report our experience with and the outcomes of PLDLH including the middle hepatic vein (MHV) and to compare these outcomes with conventional donor left hepatectomy (CDLH). The medical records of living liver donors between January 2010 and January 2018 at Seoul National University Hospital were retrospectively reviewed. Donors who underwent left hepatectomy including the MHV were included. To minimize selection bias, donors who underwent CDLH after the initiation of the PLDH program were excluded. Finally, there were 18 donors who underwent CDLH and 8 who underwent PLDLH. The median (interquartile range [IQR]) warm ischemia time (11 [10-16] minutes versus 4 [2-7] minutes; P = 0.001) was longer in the PLDLH group than the CDLH group. The total operation time (333 [281-376] minutes versus 265 [255-308] minutes; P = 0.09) and time to remove the liver (245 [196-276] minutes versus 182 [172-205] minutes; P = 0.08) were also longer in PLDLH although not statistically significant. The length of postoperative hospital stay was significantly shorter in the PLDLH group (7 [7-8] days versus 9 [8-10] days; P = 0.01). There were no postoperative complications in the PLDLH group. The rate of complications in recipients was similar in both groups. In conclusion, PLDLH including the MHV appears to be safe and feasible. Further analysis including longterm outcomes is needed.

[Effects of a Daily Life-Based Physical Activity Enhancement Program for Middle-Aged Women at Risk for Cardiovascular Disease].

PURPOSE: The purpose of this study was to examine the effects of a daily life-based physical activity enhancement program performed by middle-aged women at risk for cardiovascular disease. METHODS: This study used a randomized control group pretest-posttest design. Middle-aged women aged 45 to 64 were recruited from two outpatient cardiology departments, and randomly assigned to an experimental group (n=28) and a control group (n=30). For the experimental group, after providing one-on-one counseling and education, we provided customized text messages to motivate them in daily life. To monitor the practice of physical activity, they also used an exercise diary and mobile pedometer for 12 weeks. Subjects' physical activities (MET-min/week) were measured using the International Physical Activity Questionnaire (IPAQ). Their physiological data were obtained by blood tests using a portable analyzer, and the data were analyzed using the SPSS 21.0/WIN program. RESULTS: There were significant differences in exercise self-efficacy, health behavior, IPAQ score, body fat, body muscle, and fasting blood sugar between the two groups. However, there were no significant differences in total cholesterol, hemoglobin A1c, high-density lipoprotein cholesterol, and waist-to-hip ratio. CONCLUSION: Strengthening physical activity in daily life without being limited by cost burden and time and space constraints. Therefore, it is essential to motivate middle-aged women at risk for cardiovascular disease to practice activities that are easily performed in their daily lives.

Grape Seed Extract Supplementation Attenuates the Blood Pressure Response to Exercise in Prehypertensive Men.

We tested the hypothesis that exaggerated pressor responses observed in prehypertensive males (N = 9) during dynamic exercise are attenuated following acute dietary supplementation with grape seed extract (GSE) (i.e., a single dose). Effects of placebo and GSE (300 mg) on systolic blood pressure, diastolic blood pressure, mean arterial pressure (MAP), cardiac output (CO), stroke volume (SV), total vascular conductance (TVC), and rate × pressure product (RPP) in response to two submaximal cycling workloads (40% and 60% VO2peak) were compared 2 h after ingestion of GSE or placebo on different days, 1 week apart. Endothelial function was also evaluated using flow-mediated dilation (FMD). Placebo treatment had no effect on any of the variables. GSE supplementation attenuated MAP at both workloads (40% VO2peak: 115 ± 1 vs. 112 ± 2 mmHg; 60% VO2peak: 126 ± 2 vs. 123 ± 2 mmHg) and RPP at the lower workload. Conversely, SV, CO, and TVC were augmented during both workloads. FMD was augmented by GSE (18.9 ± 2.0 vs. 12.4% ± 2.0%). These findings indicate that in exercising prehypertensive males, a single dose of GSE reduces blood pressure, peripheral vasoconstriction, and work of the heart and enhances O2 delivery; effects that may be due, in part, to endothelium-dependent vasodilation. We propose that acute GSE treatment represents an intervention that may minimize potential increases in the risk of cardiovascular events during dynamic exercise in prehypertensives.

CD44-shRNA recombinant adenovirus inhibits cell proliferation, invasion, and migration, and promotes apoptosis in HCT116 colon cancer cells.

The cell-surface glycoprotein CD44 is closely associated with cell proliferation, tumor invasion, and metastasis. Previous studies have reported that knockdown of CD44 with short hairpin RNA (shRNA) reduced cell proliferation and migration, and induced apoptosis. However, more efficient means of delivering small interference RNA are still necessary. We developed an in vitro model of CD44-shRNA recombinant adenovirus (Ad-CD44-shRNA) and evaluated its ability to alter tumor invasion, migration, and apoptosis in human colon cancer cells. An shRNA against CD44 was used for knockdown of CD44 expression, and recombinant adenovirus was constructed using AD293 cells. The Ad-CD44-shRNA-treated HCT116 colon cancer cells showed a significant decrease in cell proliferation, migration, and invasion, while apoptosis was increased. The Ad-CD44-shRNA also decreased the phosphorylation of Akt and GSK-3ß. The levels of Bcl-2 and Bcl-xL expression were downregulated, whereas the expression levels of Bax, cleaved caspase­3 and -9, and PARP were increased in Ad-CD44-shRNA-treated colon cancer cells. These results support the feasibility of an adenovirus-mediated RNA interference therapy targeting human colon cancer via the CD44 as a potential future therapeutic intervention.

The role of metaboreceptor on exercise in hyperthermic environment with college basketball players.

The objective of this study is to review physiological differences of college basketball players cardiovascular responses and group IV metaboreceptor interactions appearing post muscular ischemia exercise (PEMI) caused by a static handgrip exercise (SHE). The subjects were placed in a temperature and moisture stabilized indoor environment for 2 h in order to measure blood pressure. For the SHE, maximal voluntary contraction of arms with a relative strength of 50 % of the maximum muscular strength was put into isometric training for 2 min. After completing the exercises, cuffs worn on the arms of the subjects were pressurized up to 200 mmHg by applying PEMI to block the artery and vein. In this way, the cardiovascular responses created by SHE and PEMI were measured. Blood samples of subjects were collected from the vein of each upper arm before SHE and after PEMI to measure the metabolite hormone and catecholamine in the blood. Results from the measurements showed a significant decrease of blood pressure under high temperature environments compared to normal temperature environments. With respect to PEMI, increases in blood pressure under the high temperature environment were significantly lower compared to the normal temperature environment. In conclusion, this study revealed that college basketball players with good physical strength had higher sensitivities of arterial baroreceptor. However, blood pressure was not increased accordingly because the increase of cutaneous vasoconstriction due to stimuli of the metaboreceptor under a high temperature environment could not be compensated by arterial baroreflex due to the increase of total vascular conductance.

Clinical Application of the Threshold Equalizing Noise Test in Patients with Hearing Loss of Various Etiologies: A Preliminary Study.

BACKGROUND AND OBJECTIVES: We aimed to make a preliminary assessment of the prevalence of cochlear dead regions (DRs) and the factors affecting the results of the threshold-equalizing noise (TEN) test in patients with hearing loss of various etiologies. SUBJECTS AND METHODS: Between May and July 2014, 109 patients (191 ears) with hearing loss who visited our outpatient clinic were prospectively enrolled. Pure tone audiometry and TEN (HL) test were performed for all the patients. DR at each frequency was indicated by masked thresholds of ≥10 dB above the TEN level and ≥10 dB above the absolute threshold. RESULTS: DR was present in 15.7% (n=30) of the 191 ears. According to disease entity, 16.6% of patients with sensorineural hearing loss had a DR. However, DR was absent in patients with chronic otitis media. According to audiometric configurations, DR was most common in moderately severe, flat hearing loss. Significantly worse hearing thresholds for both mean hearing level and hearing threshold at each frequency were found in the presence of DR (p<0.001). Logistic regression analysis showed that only the mean hearing level (odds ratio: 1.053, 95% confidence interval: 1.021-1.085) affected the presence of DR. CONCLUSIONS: Although performance of the TEN test is limited by frequencies and hearing levels, it provides additional information regarding DRs and may therefore have the potential to be used as a prognostic tool for diverse diseases causing hearing loss.

Mechanisms Underlying Exaggerated Metaboreflex Activation in Prehypertensive Men.

PURPOSE: Previously, we found that the pressor response to muscle metaboreflex activation is enhanced in prehypertension and associated with peripheral vasoconstriction. However, mechanisms underlying this exaggerated response are not clear. Therefore, we tested the hypothesis that activation of this reflex is augmented owing to increased production of muscle metabolites (i.e., lactate, K, and H). METHODS: Twenty-two men (11 normotensive and 11 prehypertensive) were studied. Changes in cardiac output (Q˙), mean arterial pressure (MAP), and total peripheral resistance (TPR) were compared between the two groups during static exercise (SE) and postexercise muscular ischemia (PEMI). Subjects completed 2 min of SE at 50% of maximal voluntary contraction (MVC) followed by 2 min of PEMI. Venous blood samples for determination of metabolites and hormones (catecholamines, vasopressin, and plasma renin activity) were taken from the exercising and nonexercising arm, respectively. RESULTS: Mean arterial pressure responses to SE (39 ± 3 vs 31 ± 2 mm Hg) and PEMI (24 ± 3 vs 19 ± 3 mm Hg) were significantly higher in the prehypertensive group. Increases in lactate and decreases in pH during PEMI were seen in both groups. However, changes in these variables were greater in the prehypertensive group (lactate, 50.1 ± 6.2 vs 32.8 ± 7.6 mg·dL; pH, -0.06 ± 0.02 vs -0.01 ± 0.01) (P < 0.05). Postexercise muscular ischemia did not evoke increases in hormones in either group. CONCLUSIONS: Compared to the normotensive group, the augmented pressor response to the metaboreflex in the prehypertensive group was associated with greater production of muscle metabolites that activate its afferent arm. The augmented response was not associated with activation of the vasopressin and renin-angiotensin systems and greater activation of the sympathetic nervous system was not apparent. Consequently, additional factors specific to prehypertension, such as arterial stiffness, may have been involved.

Bacillus anthracis genomic DNA enhances lethal toxin-induced cytotoxicity through TNF-α production.

BACKGROUND: Bacillus anthracis is the etiological agent of anthrax. Lethal toxin (LT) produced by B. anthracis is a well-known key virulence factor for anthrax because of its strong cytotoxic activity. However, little is known about the role of B. anthracis genomic DNA (BAG) in anthrax pathogenesis. RESULTS: We examined the effect of BAG on TNF-α production and LT-mediated cytotoxicity during B. anthracis spore infection in mouse macrophage cell lines (RAW264.7 cells and J774A.1) and BALB/c mice. Infection of RAW264.7 cells with B. anthracis spores induced TNF-α expression in a multiplicity of infection (MOI)-dependent manner, and this enhancement was attenuated by the toll-like receptor (TLR) 9 inhibitor oligodeoxynucleotide (ODN)2088. BAG led to TNF-α expression in a dose- and time-dependent manner when applied to RAW264.7 cells. TNF-α expression induced by BAG was reduced by either pretreatment with TLR9 inhibitors (ODN2088 and chloroquine (CQ)) or transfection with TLR9 siRNA. Furthermore, BAG-induced TNF-α production in TLR9(+/+) macrophages was completely abrogated in TLR9(-/-) macrophages. BAG enhanced the phosphorylation of mitogen-activated protein kinases (MAPK), and BAG-induced TNF-α expression was attenuated by pretreatment with MAPK inhibitors. A reporter gene assay and confocal microscopy demonstrated that BAG increased NF-κB activation, which is responsible for TNF-α expression. Treatment with BAG alone showed no cytotoxic activity on the macrophage cell line J774A.1, whereas LT-mediated cytotoxicity was enhanced by treatment with BAG or TNF-α. Enhanced LT-induced lethality was also confirmed by BAG administration in mice. Furthermore, LT plus BAG-mediated lethality was significantly recovered by administration of Infliximab, an anti-TNF-α monoclonal antibody. CONCLUSIONS: Our results suggest that B. anthracis DNA may contribute to anthrax pathogenesis by enhancing LT activity via TLR9-mediated TNF-α production.

Immunoproteomically identified GBAA_0345, alkyl hydroperoxide reductase subunit C is a potential target for multivalent anthrax vaccine.

Anthrax is caused by the spore-forming bacterium Bacillus anthracis, which has been used as a weapon for bioterrorism. Although current vaccines are effective, they involve prolonged dose regimens and often cause adverse reactions. High rates of mortality associated with anthrax have made the development of an improved vaccine a top priority. To identify novel vaccine candidates, we applied an immunoproteomics approach. Using sera from convalescent guinea pigs or from human patients with anthrax, we identified 34 immunogenic proteins from the virulent B. anthracis H9401. To evaluate vaccine candidates, six were expressed as recombinant proteins and tested in vivo. Two proteins, rGBAA_0345 (alkyl hydroperoxide reductase subunit C) and rGBAA_3990 (malonyl CoA-acyl carrier protein transacylase), have afforded guinea pigs partial protection from a subsequent virulent-spore challenge. Moreover, combined vaccination with rGBAA_0345 and rPA (protective antigen) exhibited an enhanced ability to protect against anthrax mortality. Finally, we demonstrated that GBAA_0345 localizes to anthrax spores and bacilli. Our results indicate that rGBAA_0345 may be a potential component of a multivalent anthrax vaccine, as it enhances the efficacy of rPA vaccination. This is the first time that sera from patients with anthrax have been used to interrogate the proteome of virulent B. anthracis vegetative cells.

Augmentation of the exercise pressor reflex in prehypertension: roles of the muscle metaboreflex and mechanoreflex.

This study investigated the hemodynamic mechanisms underlying the exaggerated blood pressure response to muscle contraction in prehypertensive humans and the potential role of skeletal muscle metabo- and mechanoreceptors in this response. To accomplish this, changes in peak mean arterial blood pressure (ΔMAP), cardiac output, and total peripheral resistance (ΔTPR) were compared between prehypertensive (n = 23) and normotensive (n = 19) male subjects during 2 min of static contraction (at 50% of maximal tension), 2 min of postexercise muscle ischemia (metaboreflex), and 1 min of passive dorsiflexion of the foot (tendon stretch, mechanoreceptor reflex). These variables were assessed before and during the interventions. Percentage increases from baseline in MAP and TPR in response to the exercise pressor reflex were augmented in the prehypertensives, compared with the normotensives (44% ± 5% vs. 33% ± 4% and 34% ± 15% vs. 2% ± 8%, respectively) (p < 0.05). Metaboreflex-induced increases in MAP and TPR were also augmented in the prehypertensives (28% ± 5% vs. 14% ± 4% and 36% ± 12% vs. 14% ± 9%, respectively) (p < 0.05). In response to the mechanoreflex, no differences in the percentage increase in MAP or TPR were seen between groups. The results indicate that the reflex pressor response to static contraction is augmented in prehypertension and suggest that this phenomenon is due, at least in part, to enhanced activation of metaboreceptors.

Skeletal muscle metaboreflex is enhanced in postmenopausal women.

This study determined whether an elevated muscle metaboreflex contributes to the excessive blood pressure response to exercise in postmenopausal women. Thirty healthy female volunteers were studied (15 postmenopausal and 15 premenopausal). Stroke volume, heart rate, cardiac output (CO), systolic blood pressure, diastolic blood pressure, and total vascular conductance (TVC) were continuously assessed throughout the experiment. To activate the muscle metaboreflex, occlusion of the vasculature was induced via inflation of a blood pressure cuff (2 min) on the upper arm following static handgrip exercise. Muscle metaboreflex activation increased mean arterial pressure (MAP) in both groups. However, this pressor response was greater in the postmenopausal women (ΔMAP: 21.4 ± 3 vs. 14.5 ± 2 mmHg) (P < 0.05) even though the corresponding increase in CO was less (ΔCO: 0.0 ± 0.2 vs. 0.3 ± 0.2 l/min) (P < 0.05). TVC decreased in both the groups but was more pronounced in the postmenopausal group (ΔTVC: -10.7 ± 2.6 vs. -17.1 ± 3.6 ml/min/mmHg) (P < 0.05). In conclusion, the exaggerated blood pressure response to exercise in postmenopausal women is mediated, in part, by an overactive metaboreflex that is associated with enhanced peripheral vasoconstriction.

Tumor suppressor BRCA1 epigenetically controls oncogenic microRNA-155.

BRCA1, a well-known tumor suppressor with multiple interacting partners, is predicted to have diverse biological functions. However, so far its only well-established role is in the repair of damaged DNA and cell cycle regulation. In this regard, the etiopathological study of low-penetrant variants of BRCA1 provides an opportunity to uncover its other physiologically important functions. Using this rationale, we studied the R1699Q variant of BRCA1, a potentially moderate-risk variant, and found that it does not impair DNA damage repair but abrogates the repression of microRNA-155 (miR-155), a bona fide oncomir. Mechanistically, we found that BRCA1 epigenetically represses miR-155 expression via its association with HDAC2, which deacetylates histones H2A and H3 on the miR-155 promoter. We show that overexpression of miR-155 accelerates but the knockdown of miR-155 attenuates the growth of tumor cell lines in vivo. Our findings demonstrate a new mode of tumor suppression by BRCA1 and suggest that miR-155 is a potential therapeutic target for BRCA1-deficient tumors.