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Background Abnormal renal hemodynamic responses to salt‐loading are thought to contribute to salt‐sensitive (SS) hypertension. However, this is based largely on studies in anesthetized animals, and little data are available in conscious SS and salt‐resistant rats. Methods and Results We assessed arterial blood pressure, renal function, and renal blood flow during administration of a 0.4% NaCl and a high‐salt (4.0% NaCl) diet in conscious, chronically instrumented 10‐ to 14‐week‐old Dahl SS and consomic SS rats in which chromosome 1 from the salt‐resistant Brown‐Norway strain was introgressed into the genome of the SS strain (SS.BN1). Three weeks of high salt intake significantly increased blood pressure (20%) and exacerbated renal injury in SS rats. In contrast, the increase in blood pressure (5%) was similarly attenuated in Brown‐Norway and SS.BN1 rats, and both strains were completely protected against renal injury. In SS.BN1 rats, 1 week of high salt intake was associated with a significant decrease in renal vascular resistance (−8%) and increase in renal blood flow (15%). In contrast, renal vascular resistance failed to decrease, and renal blood flow remained unchanged in SS rats during high salt intake. Finally, urinary sodium excretion and glomerular filtration rate were similar between SS and SS.BN1 rats during 0.4% NaCl and high salt intake. Conclusions Our data support the concept that renal vasodysfunction contributes to blood pressure salt sensitivity in Dahl SS rats, and that genes on rat chromosome 1 play a major role in modulating renal hemodynamic responses to salt loading and salt‐induced hypertension.

Objectives To define the magnitude of buprenorphine presence in the urine drug screens of pregnant women and to assess the presence of illicit buprenorphine use versus the presence of prescribed buprenorphine use. Methods Initial prenatal drug screen results for all pregnant patients in our practice for a 1-year period were analyzed and tabulated. Results Buprenorphine was found in the urine drug screens of 16% of pregnant patients. The presence of buprenorphine was by far the highest for any substance associated with neonatal abstinence syndrome (NAS). We estimate that the exposure to buprenorphine of approximately one-third of individuals in our population is associated with illicit buprenorphine use. Conclusions The high rate of NAS in our region is primarily associated with both illicit and prescribed buprenorphine rather than other substances. Buprenorphine usage at the time that prenatal care is initiated, rather than opiate use at the onset of prenatal care, is the underlying factor that must be addressed if our region is to successfully combat our high rates of NAS.

Bazyler, CD, Mizuguchi, S, Zourdos, MC, Sato, K, Kavanaugh, AA, DeWeese, BH, Breuel, KF, and Stone, MH. Characteristics of a national level female weightlifter peaking for competition: A case study. J Strength Cond Res 32(11): 3029-3038, 2018-This study investigated physiological and performance changes of a national-level 69 kg female weightlifter after 3 competition phases over a 28-week training period. The athlete first trained for a regional championship (weeks 1-12), followed by a local competition (weeks 13-23) and the national championship (weeks 24-28). Body mass, vastus lateralis cross-sectional area (CSA), and unloaded and loaded squat jump performance were assessed weekly during each 4-week competition phase. Serum biomarkers and dynamic midthigh pulls were assessed before and after each competition phase. Weightlifting performance goals were met for the regional championship (total = 200 kg) and the local competition (total = 193 kg), but not the national championship (total = 196 kg). She lost more body mass in preparation for Nationals (-6.0 kg) compared with regionals (-2.5 kg) and the local competition (+2.2 kg). Vastus lateralis CSA very likely decreased after Nationals (precision = 99%, effect size = 2.08). Her testosterone:cortisol ratio likely increased (88%, 2.64), whereas interleukin-6 (79%, 2.47) and tumor necrosis factor-alpha (81%, 3.59) likely decreased after Nationals. Serum myostatin (99%, 1.95) and decorin (99%, 1.96) very likely decreased after the local competition. Unloaded squat jump height likely increased the week of regionals (89%, 0.95) and the local competition (99%, 1.83), whereas unloaded and loaded squat jump height possibly (69%, 0.99) and likely (82%, 1.52) decreased the week of Nationals. Dynamic midthigh pull vertical displacement likely increased after regionals (93%, 0.84) and likely decreased after Nationals (94%, 0.87). These findings indicate that biomarkers of stress, inflammation, and hypertrophy are related to changes in training volume-load; however, performance measures are needed to assess competition preparedness. Considering the reductions in muscle CSA corresponding with the large reductions in body mass and underperformance at the national championship, sport scientists, and coaches should instruct weightlifters to not attempt large losses in body mass (e.g., >3 kg) close to competition (e.g.

We previously reported that Prunella vulgaris aqueous extract (PVAE) promotes hepatic glycogen synthesis and decreases postprandial hyperglycemia in ICR mice. Inflammatory cytokines play a critical role in the pathogenesis of diabetes. This study was designed to examine whether PVAE has a protective effect on IL-1β-induced apoptosis in INS-1 cells. INS-1 pancreatic β cells were plated at 2×10(6)/ml and treated with PVAE (100 µg/ml) 30 min before the cells were challenged with IL-1β (10 ng/ml). Untreated INS-1 cells served as control. INS-1 cell cytotoxicity was examined by MTT and lactate dehydrogenase (LDH) activity assays. Caspase-3 activity and activation of the apoptotic signaling pathway were analyzed by western blotting. NF-κB binding activity was examined by EMSA. The levels of inflammatory cytokines in the supernatant were measured by ELISA. IL-1β treatment significantly induced INS-1 cell death by 49.2%, increased LDH activity by 1.5-fold and caspase-3 activity by 7.6-fold, respectively, compared with control cells. However, PVAE administration significantly prevented IL-1β-increased INS-1 cell death and LDH activity and attenuated IL-1β-increased caspase-3 activity. Western blot data showed that PVAE also significantly attenuated IL-1β-increased Fas, FasL and phospho-JNK levels in the INS-1 cells. In addition, PVAE treatment significantly attenuated IL-1β-increased NF-κB binding activity and prevented IL-1β-increased TNF-α and IL-6 expression in INS-1 cells. Our data suggest that PVAE has a protective effect on IL-1β-induced INS-1 cell apoptosis. PVAE also attenuates IL-1β-increased NF-κB binding activity and inflammatory cytokine expression in INS-1 cells. PVAE may have a benefit for type I diabetic patients.

Environmental and anthropogenic stressors can affect wildlife populations in a number of ways. For marine mammals (e.g. the Florida manatee Trichechus manatus latirostris), certain stres- sors or conservation risk factors have been identified, but sublethal effects have been very difficult to assess using traditional methods. The development of 'biomarkers' allows us to correlate effects, such as impaired reproduction, with possible causes. A recently developed biomarker (anti-Mullerian hor- mone, AMH) provides an enzyme-linked immunosorbent assay of gonadal function. The study objec- tive was to determine AMH levels in wild manatees. In total, 28 male and 17 female manatee serum samples were assayed. Animal demographics included collection date, body weight (kg) and total length (cm). In certain cases, age of individuals was also known. AMH levels ranged from 160 to 2451.85 ng ml -1 (mean = 844.65 ng ml -1 ) in males and 0.00 to 0.38 ng ml -1 (mean = 0.10 ng ml -1 ) in females. Linear regression analyses revealed a significant relationship between male AMH levels and body weight (R 2 = 0.452; p < 0.001) and length (R 2 = 0.338; p < 0.001). Due to the small sample size, regression analyses for female AMH and body weight and length were not significant. This rep- resents the first report of AMH detection in a marine mammal. AMH levels in male manatees are the highest of any species observed to date, whereas levels in females are within reported ranges. Fur- ther studies will promote improved conservation decision by assessing AMH levels in the manatee as a function of various stressors including, but not limited to, nutritional status, serious injuries (e.g. watercraft collisions), exposure to biotoxins or contaminants, or disease.

Eicosanoid production by macrophages is an early response to microbial infection that promotes acute inflammation. The intracellular pathogen Listeria monocytogenes stimulates arachidonic acid release and eicosanoid production from resident mouse peritoneal macrophages through activation of group IVA cytosolic phospholipase A2 (cPLA2alpha). The ability of wild type L. monocytogenes (WTLM) to stimulate arachidonic acid release is partially dependent on the virulence factor listeriolysin O; however, WTLM and L. monocytogenes lacking listeriolysin O (DeltahlyLM) induce similar levels of cyclooxygenase 2. Arachidonic acid release requires activation of MAPKs by WTLM and DeltahlyLM. The attenuated release of arachidonic acid that is observed in TLR2-/- and MyD88-/- macrophages infected with WTLM and DeltahlyLM correlates with diminished MAPK activation. WTLM but not DeltahlyLM increases intracellular calcium, which is implicated in regulation of cPLA2alpha. Prostaglandin E2, prostaglandin I2, and leukotriene C4 are produced by cPLA2alpha+/+ but not cPLA2alpha-/- macrophages in response to WTLM and DeltahlyLM. Tumor necrosis factor (TNF)-alpha production is significantly lower in cPLA2alpha+/+ than in cPLA2alpha-/- macrophages infected with WTLM and DeltahlyLM. Treatment of infected cPLA2alpha+/+ macrophages with the cyclooxygenase inhibitor indomethacin increases TNFalpha production to the level produced by cPLA2alpha-/- macrophages implicating prostaglandins in TNFalpha down-regulation. Therefore activation of cPLA2alpha in macrophages may impact immune responses to L. monocytogenes.

Glucans are fungal cell wall glucose polymers that are released into the blood of infected patients. The role of glucans in infection is unknown. We examined serum glucan and cytokine levels in intensive care unit (ICU) patients with infections. There was an inverse correlation (p0.001) between serum glucan levels and interleukin (IL)-2), IL-4, tumor necrosis factoralpha (TNFalpha) and granulocyte macrophage-colony stimulating factor (GM-CSF) levels in infected ICU patients. The correlation between serum cytokines and serum glucan was only observed at glucan concentrations40 pg/ml. No change was observed at serum glucan levels of40 pg/ml. There was no correlation between serum glucan levels and systemic levels of IL-1beta, IL-5, IL-6, IL-8, IL-10 or IFNgamma. Interestingly, blood borne glucans did not suppress systemic cytokine levels in infected ICU patients, instead they were maintained at control levels. We conclude that circulating glucans may prevent cytokine upregulation in response to infection. This may represent an adaptive response to septic injury.

Objective: Determine if aspirin inhibits the IL-1β–stimulated expression of inducible nitric oxide synthase (iNOS), nitric oxide (NO), and prostaglandin E 2 (PGE 2 ) in rat ovarian dispersates cultured in vitro . Design: Prospective, controlled in vitro study. Setting: Academic research laboratory. Animals: Ovaries collected from immature rats. Intervention(s): Ovaries were collected from immature rats and enzymatically dispersed. Ovarian dispersates were placed into plates containing media alone or media supplemented with IL-1β (100 U/mL) and varying concentrations of aspirin (0, 1, 3, 5 and 10 mM). Ovarian dispersates were cultured in a humidified environment of 5% CO 2 in air at 37°C for 24 or 48 hours. Main Outcome Measure(s): Twenty-four– and 48-hour iNOS, nitrite (a stable metabolite of NO), and PGE 2 levels were determined from ovarian dispersates cultured in vitro. Result(s): Administration of IL-1β increased nitrite and PGE 2 levels over that observed in the control group after culture of ovarian dispersates for 24 and 48 hours. Aspirin dose dependently reduced the IL-1β–stimulated increase in nitrite production from ovarian dispersates after culture for 24 and 48 hours. Aspirin completely (24 hours) or dose dependently (48 hours) prevented the IL-β–stimulated increase in PGE 2. Coadministration of IL-1β and aspirin (10 mM) attenuates IL-1β–stimulated iNOS expression after culture for 24 and 48 hours. Conclusion(s): Aspirin significantly inhibits the IL-1β–stimulated expression of iNOS, NO, and PGE 2 in ovarian dispersates cultured in vitro.

Platelet activating factor (PAF) was administered to female mice in order to investigate its effect on ovulation rate and on oocyte quality including their in-vitro embryonic development, implantation and uterine receptivity. In experiment 1, 4-week-old female mice were assigned to receive PAF or phosphate buffered saline for 4 consecutive days. On the second day of this treatment, pregnant mares' serum gonadotrophin was administered and human chorionic gonadotrophin (HCG) 48 h later, after which copulation occurred. Oocytes were collected on the following day and evaluated. The mean number of oocytes and zygotes (two pronuclear stage embryos) recovered from the PAF-treated group was not different from the control group (31 versus 27), but the proportion of zygotes was higher in PAF-treated group than in controls (83 versus 68%, P C: n = 19), control embryo to PAF recipient (C-->PAF: n = 19), and control embryo to control recipient (C-->C: n = 22). Implantation and abortion were assessed on day 19 posttransfer. The implantation rate of C-->PAF (23.7%) was lower than C-->C (31.1%, P C (31.2%). Further, C-->PAF showed a higher abortion rate per embryo (29.6%) than PAF-->C (12.7%, P C (24.4%). In the present study, PAF administration enables females to produce oocytes with a higher potential for fertilization, in-vitro development and implantation, but has a detrimental effect on uterine receptivity to embryos.

Phosphoinositide-3-kinase (PI3K)/Akt dependent signaling has been shown to improve outcome in sepsis/septic shock. There is also ample evidence that PI3K/Akt dependent signaling plays a crucial role in maintaining normal cardiac function. We hypothesized that PI3K/Akt signaling may ameliorate septic shock by attenuating sepsis-induced cardiac dysfunction. Cardiac function and survival were evaluated in transgenic mice with cardiac myocyte specific expression of constitutively active PI3K isoform, p110α (caPI3K Tg). caPI3K Tg and wild type (WT) mice were subjected to cecal ligation/puncture (CLP) induced sepsis. Wild type CLP mice showed dramatic cardiac dysfunction at 6 hrs. Septic cardiomyopathy was significantly attenuated in caPI3K CLP mice. The time to 100% mortality was 46 hrs in WT CLP mice. In contrast, 80% of the caPI3K mice survived at 46 hrs after CLP (p30 days (p

Sepsis is a frequent complication in critical illness. The mechanisms that are involved in initiation and propagation of the disease are not well understood. Scavenger receptor A (SRA) is a membrane receptor that binds multiple polyanions such as oxidized LDL and endotoxin. Recent studies suggest that SRA acts as a pattern recognition receptor in the innate immune response. The goal of the present study was to determine the role of SRA in polymicrobial sepsis. SRA deficient (SRA−/−) and C57BL/6JB/6J (WT) male mice were subjected to cecal ligation and puncture (CLP) to induce polymicrobial sepsis. NFκB activity, myeloperoxidase activity, and co-association of SRA with toll like receptor (TLR) 4 and TLR2 was analyzed in the lungs. Spleens were analyzed for apoptosis. Serum cytokines and chemokines were assayed. Blood and peritoneal fluid were cultured for aerobic and anaerobic bacterial burdens. Long term survival was significantly increased in SRA−/− septic mice (53.6% vs. 3.6%, p, Author Summary Trauma and other critical illnesses can progress to septic shock. The mechanisms that result in this progression are not understood. For this reason, there are no proven treatments available, and the mortality rate from sepsis remains quite high. We have found that mice that lack a certain cell surface protein, scavenger receptor A, have a higher rate of survival from a surgically induced sepsis than those that have the receptor. Previously, this receptor has been found to play a role in atherosclerosis, and more recently, to play a role in the immune response to infection. In this study we have found that in addition to improved survival, mice without scavenger receptor A have fewer bacteria in their abdominal cavities and in their blood. They also have lower levels of inflammation. We demonstrated that scavenger receptor A interacts with another protein involved in inflammation and infection, toll like receptor 4. This interaction might be one mechanism for the effects seen in mice without scavenger receptor A. These studies provide a better understanding of the underlying mechanisms of sepsis. Drugs that target scavenger receptor A could result in better therapies for sepsis.

Heterotrimeric G(i) proteins play a role in signalling activated by lipopolysaccharide (LPS), Staphylococcus aureus (SA) and group B streptococci (GBS), leading to production of inflammatory mediators. We hypothesized that genetic deletion of G(i) proteins would alter cytokine and chemokine production induced by LPS, SA and GBS stimulation. LPS-induced, heat-killed SA-induced and heat-killed GBS-induced cytokine and chemokine production in peritoneal macrophages from wild-type (WT), Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice were investigated. LPS induced production of tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), IL-10 and interferon-gamma-inducible protein-10 (IP-10); SA induced TNF-alpha, and IL-1beta production; and GBS induced TNF-alpha, IL-6, IL-1beta, macrophage inflammatory protein-1alpha (MIP-1alpha) and keratinocyte chemoattract (KC) production were all decreased (P0.05) in Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice compared with WT mice. In contrast to the role of G(i) proteins as a positive regulator of mediators, LPS-induced production of MIP-1alpha and granulocyte-macrophage colony-stimulating factor (GM-CSF) were increased in macrophages from Galpha(i1/3) (-/-) mice, and SA-induced MIP-1alpha production was increased in both groups of Galpha(i) protein-depleted mice. LPS-induced production of KC and IL-1beta, SA-induced production of GM-CSF, KC and IP-10, and GBS-induced production of IL-10, GM-CSF and IP-10 were unchanged in macrophages from Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice compared with WT mice. These data suggest that G(i2) and G(i1/3) proteins are both involved and differentially regulate murine inflammatory cytokine and chemokine production in response to both LPS and Gram-positive microbial stimuli.

Heterotrimeric Gi proteins play a role in lipopolysaccharide (LPS) and Staphylococcus aureus (SA) activated signaling leading to inflammatory mediator production. We hypothesized that genetic deletion of Gi proteins would alter cytokine and chemokine production induced by LPS and SA. LPS- and heat killed SA-induced cytokine and chemokine production in splenocytes from wild type (WT), Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice were investigated. LPS- or SA-induced production of TNFalpha, IL-6, IFNgamma, IL-12, IL-17, GM-CSF, MIP-1alpha, MCP-1, MIG and IP-10 were significantly increased (1.2 to 33 fold, p0.05) in splenocytes harvested from Galpha(i2)(-/-) mice compared with WT mice. The effect of Galpha(i) protein depletion was remarkably isoform specific. In splenocytes from Galpha(i1/3) (-/-) mice relative to WT mice, SA-induced IL-6, IFNgamma, GM-CSF, and IP-10 levels were decreased (59% to 86%, p0.05), whereas other LPS- or SA-stimulated cytokines and chemokines were not different relative to WT mice. LPS- and SA-induced production of KC were unchanged in both groups of the genetic deficient mice. Splenocytes from both Galpha(i2) (-/-) and Galpha(i1/3) (-/-) mice did not exhibit changes in TLR2 and TLR4 expression. Also analysis of splenic cellular composition by flow cytometry demonstrated an increase in splenic macrophages and reduced CD4 T cells in both Galpha(i2) (-/-) and Galpha(i1/3) (-/-) mice relative to WT mice. The disparate response of splenocytes from the Galpha(i2) (-/-) relative to Galpha(i1/3) (-/-) mice therefore cannot be attributed to major differences in spleen cellular composition. These data demonstrate that G(i2) and G(i1/3) proteins are both involved and differentially regulate splenocyte inflammatory cytokine and chemokine production in a highly Gi isoform specific manner in response to LPS and Gram-positive microbial stimuli.

Mast cells are highly responsive cells that are capable of secreting a variety of inflammatory mediators, including histamine, heparin, serine proteases, leukotrienes, prostaglandins, and thromboxanes. Studies from several laboratories have demonstrated that mast cells have the capacity to produce a variety of cytokines in response to various stimuli. Characterization of the cytokine profiles in mast cells has routinely been determined by the performance of individual enzyme-linked immunosorbent assays. This process is expensive, time-consuming, and requires a great deal of material to characterize multiple cytokines. In this chapter, we describe a multiplex cytokine assay to detect 17 cytokines simultaneously in 50 microL of culture supernatant derived from stimulated human cord blood-derived mast cells.

Mast cells are highly responsive cells that are capable of secreting a variety of inflammatory mediators, including histamine, heparin, serine proteases, leukotrienes, prostaglandins, and thromboxanes. Studies from several laboratories have demonstrated that mast cells have the capacity to produce a variety of cytokines in response to various stimuli. Characterization of the cytokine profiles in mast cells has routinely been determined by the performance of individual enzyme-linked immunosorbent assays. This process is expensive, time-consuming, and requires a great deal of material to characterize multiple cytokines. In this chapter, we describe a multiplex cytokine assay to detect 17 cytokines simultaneously in 50 microL of culture supernatant derived from stimulated human cord blood-derived mast cells.

Two experiments were performed to determine whether administration of ergotamine tartrate altered embryo development (Exp. 1) and uterine competency to establish pregnancy (Exp. 2) in beef cattle. Animals were fed daily either 0 (CON) or 40 mug/kg body weight of ergotamine tartrate (ERGOT). Following a 30-d period on respective diets, animals in Exp. 1 were artificial inseminated at estrus (d = 0) and single embryo recoveries performed on day 7; whereas, animals in Exp. 2 received two frozen-thawed embryos on day 7. As an indicator of ergotamine effects, prolactin was decreased throughout both experiments in ERGOT compared to CON animals (p0.05). Furthermore, rectal temperature (RT) tended to increase during both experiments in ERGOT compared to CON (p= 0.06). In Exp. 1, embryo recovery (p=0.08) and the percentage of transferable embryos (p=0.09) tended to be greater for CON than for ERGOT. Percentage of embryos that developed to compacted morula or greater was increased for CON compared to ERGOT heifers (p0.05). In Exp. 2, pregnancy rates of transferred embryos did not differ between treatment groups. Thus, administration of ERGOT to simulate fescue toxicosis altered developmental potential of embryos, but does not appear to affect uterine competency to establish pregnancy.

LPS pretreatment of human pro-monocytic THP-1 cells induces tolerance to secondary LPS stimulation with reduced TNFalpha production. However, secondary stimulation with heat-killed Staphylococcus aureus (HKSa) induces priming as evidenced by augmented TNFalpha production. The pro-inflammatory cytokine, IFNgamma, also abolishes suppression of TNFalpha in LPS tolerance. The effect of LPS tolerance on HKSa and IFNgamma-induced inflammatory mediator production is not well defined. We hypothesized that LPS, HKSa and IFNgamma differentially regulate pro-inflammatory mediators and chemokine production in LPS-induced tolerance. THP-1 cells were pretreated for 24 h with LPS (100 ng/ml) or LPS (100 ng/ml) + IFNgamma (1 microg/ml). Cells were subsequently stimulated with LPS or HKSa (10 microg/ml) for 24 h. The production of the cytokines TNFalpha, IL-6, IL-1beta, and GMCSF and the chemokine IL-8 were measured in supernatants. LPS and HKSa stimulated TNFalpha (3070 +/- 711 pg/ml and 217 +/- 9 pg/ml, respectively) and IL-6 (237 +/- 8.9 pg/ml and 56.2 +/- 2.9 pg/ml, p0.05, n = 3, respectively) in control cells compared to basal levels (25 pg/ml). LPS induced tolerance to secondary LPS stimulation as evidenced by a 90% (p0.05, n = 3) reduction in TNFalpha. However, LPS pretreatment induced priming to HKSa as demonstrated by increased TNFalpha (2.7 fold, from 217 to 580 pg/ml, p0.05, n = 3 ). In contrast to suppressed TNFalpha, IL-6 production was augmented to secondary LPS stimulation (9 fold, from 237 to 2076 pg/ml, p0.01, n = 3) and also primed to HKSa stimulation (62 fold, from 56 to 3470 pg/ml, p0.01, n = 3). LPS induced IL-8 production and to a lesser extent IL-1beta and GMCSF. LPS pretreatment did not affect secondary LPS stimulated IL-8 or IL-1beta, although HKSa stimulation augmented both mediators. In addition, IFNgamma pretreatment reversed LPS tolerance as evidenced by increased TNFalpha levels while IL-6, IL-1beta, and GMCSF levels were further augmented. However, IL-8 production was not affected by IFNgamma. These data support our hypothesis of differential regulation of cytokines and chemokines in gram-negative- and gram-positive-induced inflammatory events. Such changes may have implications in the pathogenesis of polymicrobial sepsis.

Objectives: Hormones and cytokines are known to act as regulatory messengers between the neuroendocrine and immune systems. The innate immune system identifies infectious agents by means of pattern-recognition receptors. These receptors recognize pathogen-specific macromolecules called pathogen-associated molecular patterns. Fungal cell wall glucans nonspecifically stimulate various aspects of innate immunity via interaction with membrane receptors on immune-competent cells. Glucans are also released into the systemic circulation of patients with fungal infections. Recent evidence confirms the existence of glucan-specific receptors on cells outside the immune system. We hypothesized that glucans may directly interact with pituitary cells as an early signaling event in fungal infections. Methods: We characterized the receptor-mediated interaction of glucan derived from Candida albicans with pituitary cells using surface plasmon resonance. Prolactin levels were assayed by commercial ELISA. TLR2, TLR4 and CD14 mRNA levels were assessed by RT-PCR. Results: A single glucan-specific binding site was identified on rodent somatomammotroph (KD = 3.9 µM) and human folliculostellate cell (KD = 3.6 µM) membranes. Coincubation of glucan with somatomammotroph cells for 72 h significantly (p < 0.01) increased prolactin accumulation by 56–62% over that observed in cells treated with media alone. Glucan also increased TLR4 and CD14 gene expression in human folliculostellate cells. Conclusions: Pituitary cells directly recognize and respond to fungal cell wall glucans resulting in stimulation of pituitary cell TLR4 and CD14 gene expression. In addition, glucan stimulates secretion of prolactin, a hormone that plays an important role in the response to infection.

Background Since the passage of The Dietary Supplement Health and Education Act in 1994, there has been a flood of new "dietary" supplements promoting anti-aging benefits such as the enhancement of growth hormone or testosterone levels. Androstenediol and androstenedione are such products. This study's purpose was to elucidate the physiological and hormonal effects of 200 mg/d of oral androstenediol and androstenedione supplementation in men aged 35 to 65 years while participating in a 12-week high-intensity resistance training program. Methods Fifty men not consuming any androgenic-enhancing substances and with normal total testosterone levels, prostate-specific antigen, hemoglobin, and hematocrit, and with no sign of cardiovascular or metabolic diseases participated. Subjects were randomly assigned to a placebo, androstenediol (diol), or androstenedione (dione) group using a double-blind study design. Main outcomes included serum sex hormone profile, body composition assessment, muscular strength, and blood lipid profiles. Results During the 12 weeks of androstenedione or androstenediol use, a significant increase in the aromatization by-products estrone and estradiol was observed in both groups ( P = .03). In the dione group, total testosterone levels significantly increased 16% after 1 month of use, but by the end of 12 weeks, they returned to pretreatment levels. This return to baseline levels resulted from increases in aromatization and down-regulation in endogenous testosterone synthesis based on the fact that luteinizing hormone was attenuated 18% to 33% during the treatment period. Neither androstenediol nor androstenedione enhanced the adaptations to resistance training compared with placebo for body composition or muscular strength. However, both androstenediol and androstenedione supplementation adversely affected high-density lipoprotein cholesterol (HDL-C) levels, coronary heart disease risk (representing a 6.5% increase), and each group's respective (low-density lipoprotein cholesterol [LDL-C]/HDL-C)/(apolipoprotein A/apolipoprotein B) lipid ratio (diol: +5.2%; dione: +10.5%; P = .05). In contrast, the placebo group's HDL-C levels increased 5.1%, with a 12.3% decline in the (LDL-C/HDL-C)/(apolipoprotein A/apolipoprotein B) lipid ratio. These negative and positive lipid effects occurred despite no significant alterations in body composition or dietary intakes in the supplemental groups or placebo group, respectively. Conclusions Testosterone precursors do not enhance adaptations to resistance training when consumed in dosages recommended by manufacturers. Testosterone precursor supplementation does result in significant increases in estrogen-related compounds, dehydroepiandrosterone sulfate concentrations, down-regulation in testosterone synthesis, and unfavorable alterations in blood lipid and coronary heart disease risk profiles of men aged 35 to 65 years.

To determine if known androgenic hormone precursors for testosterone in the androgen pathway would be readily transformed to testosterone, eight male subjects [mean age 23.8 (SEM 3) years, bodymass 83.1 (SEM 8.7) kg, height 175.6 (SEM 8.5) cm] underwent a randomized, double-blind, cross-over, placebo-controlled oral treatment with 200 mg of 4-androstene-3,17-dione (delta 4), 4-androstene-3 beta,17 beta-diol (delta 4 Diol), and placebo (PL). The periods of study were separated by 7 days of washout. Blood was drawn at baseline and subsequently every 30 min for 90 min after treatment. Analysis revealed mean area-under-the-curve (AUC) serum delta 4 concentrations to be higher during delta 4 treatment [2177 (SEM 100) nmol.l-1] than delta 4Diol [900 (SEM 96) nmol.l-1] or PL [484 (SEM 82) nmol.l-1; P0.0001]. The delta 4 treatment also revealed a significant effect on total testosterone with a mean AUC [1632.5 (SEM 121) nmol.l-1] that was greater than PL [1418.5 (SEM 131) nmol.l-1; P0.05] but not significantly different from those observed after delta 4Diol treatment [1602.9 (SEM 119) nmol.l-1; P = 0.77]. Free testosterone concentrations followed a similar pattern where mean AUC for the delta 4 treatment [6114.0 (SEM 600) pmol.l-1] was greater than after PL [4974.6 (SEM 565) pmol.l-1; P0.06] but not significantly different from those observed after delta 4Diol [5632.0 (SEM 389) pmol.l-1; P = 0.48]. The appearance and apparent conversion to total and free testosterone over 90 min was stronger for the delta 4 treatment (r = 0.91, P0.045) than for delta 4Diol treatment (r = 0.69, NS) and negatively correlated for PL (r = -0.90, P0.02). These results would suggest that delta 4, and perhaps delta 4Diol, taken by month are capable of producing in vivo increases in testosterone concentrations in apparently healthy young men as has already been observed in women after treatment with delta 4.

BACKGROUND Rubella and congenital rubella syndrome remain a problem in the United States; institutionalized individuals are at higher risk. We assessed demographic variables and rubella serology in a population of institutionalized adults with mental retardation. METHODS Subjects were 181 institutionalized adults with mental retardation. We reviewed charts to determine patient's age, sex, race, and fertility status (if female), cause of mental retardation, and rubella history. Serologic testing to determine rubella immunity was done. RESULTS We found that 26.4% of subjects were nonimmune. Sex, cause of mental retardation, and history of rubella vaccination were not related to serologic immunity status. Of the 29 fertile female subjects, 8 (27.6%) were nonimmune to rubella. CONCLUSION Subjects with unknown immunization status had a similar serologic profile to those whose medical records indicated immunization. Neither history of infection nor immunization was predictive of serology status.

Previous studies have demonstrated that bacterial lipopolysaccharide (LPS) and heat killed Staphylococcus aureus (SA) activation of inflammatory cells depended in part upon activation of heterotrimeric Gi proteins. It has also been shown that (1 --3) beta-D-glucan can suppress inflammatory cell activation by microbial products although the cellular mechanism of the glucan effect remains to be clearly defined. We hypothesized that Gi proteins function as a common convergent signaling pathway for both LPS and SA leading to monocyte mediator production. Additionally, we hypothesized that soluble glucan suppresses LPS and SA induced cytokine production via Gi protein coupled signaling. Human THP-1 promonocytic cells were pretreated with pertussis toxin (PTx, 100 ng/ml or 1 microgram/ml) 6 hours prior to stimulation with LPS (10 microgram/ml) and SA (10 microgram/ml) and/or soluble glucan (10 microgram/ml). Both LPS and SA significantly (p0.05) induced cytokine production IL-6TNF alphaIL-1 betaGM-CSFIL-10IFN gamma. The induction of these cytokines was significantly (p0.05) suppressed by PTx. Glucan treatment alone had no effect on cytokine production but suppressed (P0.05) LPS and SA induced cytokines. PTx further augmented (p0.05) the inhibitory effect of glucan on the LPS and SA induced cytokine expression. The data support the hypothesis that Gi proteins function as a common signaling protein for both LPS and SA induction of pro-and anti-inflammatory cytokines and that soluble glucan effectively suppresses cytokine production to the microbial stimuli. In contrast, the effect of soluble glucan on inhibiting cellular activation by LPS and SA is Gi protein independent.

Background The problem of sexual abuse has been well described. It is believed that as many as 20% of women and 5–10% of men have been sexually abused as children. The perpetrator is usually a family member or other person well known to the victim. Perpetrators as young a 7 years old have been reported. Child care giver knowledge about sexual abuse has not been well described. We believe this is the first effort to describe the knowledge base of child care providers concerning the topic of sexual abuse This research will hopefully promote efforts to prevent sexual abuse. Methods A one page questionnaire examining knowledge about sexual abuse was offered to new prenatal patients presenting for care at the East Tennessee State University, Department of Obstetrics and Gynecology. Participant's (n=211) were asked questions concerning their knowledge of rates of sexual abuse among both males and females, and about perpetrator identification and age. Demographic information obtained included the participant's age, educational status and whether she was currently involved in the care of children. Participants’ interest in obtaining more information about sexual abuse was also recorded. Data were analyzed using the CATMOD procedure of SAS. Results 211 of 217 women agreed to participate in the survey. Women with greater than 12 years of education were found to be more likely (p Conclusion Sexual abuse is a topic that many new mothers are interested in learning more about. The knowledge base of these individuals can be improved. In order to prevent sexual abuse, it would be appropriate to educate child care givers about the dangers of such abuse.