Effects of Gender Difference and Caffeine Supplementation on Anaerobic Muscle Performance.

The primary purpose of this study was to investigate the effects of gender difference and caffeine supplementation to maximal voluntary isometric contractions (MVIC) and submaximal voluntary isometric contractions (T(lim)). 10 male (age: 20.10 ± 2.18 years, BMI: 22.23±1.96 kg/m(2)) and 10 female (age: 19.90±0.99 years, BMI: 21.76±2.65 kg/m(2)) elite collegiate athletes were recruited. Subjects ingested caffeine (6 mg/kg) or a placebo in a randomized, double-blind, placebo-control, and counter-balanced fashion. MVIC and T(lim) were measured after supplementations. T(lim) result was calculated based on the time to exhaustion of isometric contraction with 50% MVIC. Fatigue index (FI%) referred to the MVIC tested 20 s after the cessation of T(lim) test, and was indexed by the percentage of MVIC decline. No significant interaction effect was found between the gender factor and the supplementation factor for all dependent variables. Compared to the placebo, caffeine supplementation significantly increased MVIC (5.9%) and T(lim) (15.5%) (p<0.05) whereas it had no significant effect on FI%. This study demonstrates that caffeine supplementation at a 6 mg/kg dosage facilitates performances in MVIC and T(lim). The ergogenic effect of caffeine on muscle power and muscle endurance does not show a gender bias.

IFN-γ induces aberrant CD49b⁺ NK cell recruitment through regulating CX3CL1: a novel mechanism by which IFN-γ provokes pregnancy failure.

Interferon-γ (IFN-γ), a pleiotropic lymphokine, has important regulatory effects on many cell types. Although IFN-γ is essential for the initiation of uterine vascular modifications and maintenance of decidual integrity, IFN-γ administration can also cause pregnancy failure in many species. However, little is known about the effector mechanisms involved. In this study, using an IFN-γ-induced abortion mouse model, we reported that no Dolichos biflorus agglutinin lectin-positive uterine natural killer (uNK) cells were observed in the uteri from IFN-γ-induced abortion mice. By contrast, the percentage of CD3(-)CD49b(+) NK cells in the uterus and blood from a foetal resorption group was significantly higher than that of the control group. Similarly, significantly upregulated expression of CD49b (a pan-NK cell marker), CX3CL1 and CX3CR1 (CX3CL1 receptor) was detected in the uteri of IFN-γ-induced abortion mice. Using isolated uterine stromal cells, we showed that upregulated expression of CX3CL1 by IFN-γ was dependent on a Janus family kinase 2-signal transducers and activators of transcription 1 (JAK2-STAT1) pathway. We further demonstrated the chemotactic activity of CX3CL1 in uterine stromal cell conditioned medium on primary splenic NK cells. Finally, we observed increased recruitment of CD49b(+) NK cells into the endometrium after exogenous CX3CL1 administration. Collectively, our findings indicate that IFN-γ can significantly increase uterine CX3CL1 expression via activation of the JAK2-STAT1 pathway, thus inducing CD49b(+) NK cell uterine homing, and eventually provoke foetal loss. Thus, we provide a new line of evidence correlating the deleterious effects of IFN-γ on pregnancy with the aberrant regulation of CX3CL1 and CD49b(+) NK cells.

Error-specific medial cortical and subcortical activity during the stop signal task: a functional magnetic resonance imaging study.

The ability to detect errors and adjust behavior accordingly is essential for maneuvering in an uncertain environment. Errors are particularly prone to occur when multiple, conflicting responses are registered in a situation that requires flexible behavioral outputs. Previous studies have provided evidence indicating the importance of the medial cortical brain regions including the cingulate cortex in processing conflicting information. However, conflicting situations can be successfully resolved, or lead to errors, prompting a behavioral change in the observers. In particular, how does the brain use error signals specifically to adjust behavior on the fly? Here we employ a stop signal task (SST) to elicit errors approximately half of the time in high-conflict trials despite constant behavioral adjustment of the observers. Using functional magnetic resonance imaging, we show greater and, sequentially, less activation in the medial cortical regions when observers made an error, compared with when they successfully resolved high-conflict responses. Errors also evoked greater activity in the cuneus, retrosplenial cortex, insula, and subcortical structures including the thalamus and the region of the epithalamus (the habenula). We further showed that the error-related medial cortical activities are not correlated with post-error behavioral adjustment, as indexed by post-error slowing (PES) in go trial reaction time. These results delineate an error-specific pattern of brain activation during the SST. The results also suggest that the relationship between error-related activity and post-error behavioral adjustment may be more complicated than has been conceptualized by the conflict monitoring hypothesis.

Inhibition of angiotensin II induced endothelin-1 gene expression by 17-beta-oestradiol in rat cardiac fibroblasts.

OBJECTIVE: To examine whether 17-beta-oestradiol (E(2)) may alter angiotensin II (Ang II) induced cell proliferation and to identify the putative underlying signalling pathways in rat cardiac fibroblasts. DESIGN: Cultured rat cardiac fibroblasts were preincubated with E(2) then stimulated with Ang II. [(3)H]Thymidine incorporation and endothelin-1 (ET-1) gene expression were examined. The effect of E(2) on Ang II induced NADPH oxidase activity, reactive oxygen species (ROS) formation, and extracellular signal regulated kinase (ERK) phosphorylation were tested to elucidate the intracellular mechanism of E(2) in proliferation and ET-1 gene expression. RESULTS: Ang II increased DNA synthesis, which was inhibited with E(2) (1-100 nmol/l). E(2), but not 17-alpha-oestradiol, inhibited Ang II induced ET-1 gene expression as shown by northern blotting and promoter activity assay. This effect was prevented by co-incubation with the oestrogen receptor antagonist ICI 182,780 (1 micromol/l). E(2) also inhibited Ang II increased NADPH oxidase activity, ROS formation, ERK phosphorylation, and activator protein-1 mediated reporter activity. CONCLUSIONS: The results suggest that E(2) inhibits Ang II induced cell proliferation and ET-1 gene expression, partially by interfering with the ERK pathway through attenuation of ROS generation. Thus, this study provides important new insight regarding the molecular pathways that may contribute to the proposed beneficial effects of oestrogen on the cardiovascular system.

Female sex hormones modulate the function of LPS-treated macrophages.

PROBLEM: To study the effects of estradiol (E2) or progesterone on macrophage function in the presence of lipopolysaccharide (LPS). METHOD OF STUDY: Male rat peritoneal macrophages were treated in vitro with 0.1 microg/mL of LPS and E2 or progesterone. RESULTS: At 10(-2) ng/mL, E2 significantly (P < 0.05; n = 6) enhanced tumor necrosis factor (TNF) release by LPS-treated macrophages. TNF release was significantly (P < 0.05; n = 6) inhibited by 10(2) ng/mL or 10(3) ng/mL of E2 and by progesterone at less than 10(-3) ng/mL or greater than 10(-1) ng/mL. E2 (10(-4) and 10 ng/mL) and progesterone (10(-6)-10(-4) ng/mL and 10(2) ng/mL) each significantly (P < 0.05, n = 8) enhanced H2O2 release by LPS-treated macrophages. E2 ( < 10(-2) and > 10 ng/mL) and progesterone (10(-7)-10(4) ng/mL) each significantly inhibited (P< 0.05; n = 6) NO2- release by LPS-treated macrophages. CONCLUSIONS: Exposure to LPS tended to diminish the effects of E2 and to enhance the effects of progesterone on the parameters determined here. Such LPS-associated alterations in the dose-response profile of macrophages to female sex hormones may contribute to gender-related differences in the immune response under normal and pathological conditions.

Loss of p19(ARF) eliminates the requirement for the pRB-binding motif in simian virus 40 large T antigen-mediated transformation.

At least three domains of simian virus 40 large T antigen (TAg) participate in cellular transformation. The LXCXE motif of TAg binds to all members of the retinoblastoma protein (pRB) family of tumor suppressors. The N-terminal 70 residues of TAg have significant homology to the J domain of Hsp40/DnaJ and cooperate with the LXCXE motif to inactivate the pRB family. A bipartite C-terminal domain of TAg binds to p53 and thereby disrupts the ability of p53 to act as a sequence-specific transcription factor. The contribution of these three domains of TAg to cellular transformation was evaluated in cells that contained inactivating mutations in the pRB and p53 pathways. Cells that stably expressed wild-type or selected mutant forms of TAg were generated in mouse embryo fibroblasts (MEFs) containing homozygous deletions in the RB, INK4a, and ARF loci. It was determined that the J domain, the LXCXE motif, and the p53-binding domain of TAg were required for full transformation of wild-type and RB(-/-) MEFs. In contrast, INK4a(-/-) MEFs that lacked expression of p16(INK4a) and p19(ARF) and ARF(-/-) MEFs that lacked p19(ARF) but expressed p16(INK4a) acquired anchorage-independent growth when expressing wild-type TAg or mutant derivatives that disrupted either the pRB-binding or p53-binding domain. The expression and function of the pRB family members were not overly disrupted in ARF(-/-) MEFs expressing LXCXE mutants of TAg. These results suggest that inactivating mutations of p19(ARF) can relieve the requirement for the LXCXE motif in TAg-mediated transformation and that TAg may have additional functions in transformation.

The role of P wave in prediction of atrial fibrillation after coronary artery surgery.

Atrial fibrillation (AF) is a common arrhythmia after coronary artery bypass surgery (CABG). The purpose of this study was to determine the role of P wave duration, amplitude and dispersion in the prediction of AF after CABG. This study included 120 patients undergoing elective CABG. Clinical characteristics, 12-lead electrocardiogram (ECG), echocardiogram and coronary angiogram were obtained in all patients. We measured P wave duration, amplitude and dispersion from 12-lead ECG in each patient. After CABG, all patients were continuously monitored for AF attacks in the intensive care unit and ordinary ward. Our results showed that age greater than 60 years was the strongest predictor of postoperative AF (p<0.01), with a 3.7-fold greater likelihood of developing postoperative AF compared to ages less than 60 years. Gender was another independent predictor of postoperative AF, with men being 3.0 times more likely to develop postoperative AF compared to women (p = 0.03). The presence of prolonged P wave duration (> or =100 ms in lead II) was also an independent predictor (p = 0.04), with 2.9-fold greater risk of developing postoperative AF compared to a P wave duration of less than 100 ms. The P wave dispersion was similar between patients with and without postoperative AF (29+/-15 vs. 33+/-15 mm, p = NS). In conclusion, old age, male gender and prolonged P wave duration were independent predictors of AF after CABG. However, P wave dispersion and amplitude did not provide significant information in the prediction of postoperative AF.

Somatostatin and octreotide modulate the function of Kupffer cells in liver cirrhosis.

In our previous studies we have shown that somatostatin and octreotide modulate the function of peritoneal macrophages and Kupffer cells in noncirrhotic livers. However, the effects of somatostatin on the Kupffer cells in cirrhotic livers are not known. In the present study, Kupffer cells, obtained from male rats with carbon tetrachloride-induced cirrhotic livers, were treated in vitro with somatostatin or octreotide and their effects on the release of nitric oxide, tumor necrosis factor-alpha (TNF-alpha) and peroxide (H2O2) determined. At concentrations of 10(-13) or 10(-10) to 10(-6) M of somatostatin or 10(-12) to 10(-10) M, or 10(-6) M of octreotide, the amount of nitric oxide released by Kupffer cells was significantly suppressed relative to that of untreated cells. Kupffer cells treated with less than 10(-12) M or greater than 10(-12) M of somatostatin or octreotide released less TNF-alpha compared to the untreated controls. In addition, zymosan-induced H2O2 release by Kupffer cells treated with 10(-9) to 10(-7) M somatostatin or with 10(-15) to 10(-13) M and 10(-9) to 10(-7) M of octreotide was greater than that of the untreated controls. These findings demonstrate that somatostatin and octreotide modulate the release of nitric oxide, TNF-alpha and H2O2 by Kupffer cells in cirrhotic livers depending on the concentrations of hormones used.

Identification and characterization of GFRalpha-3, a novel Co-receptor belonging to the glial cell line-derived neurotrophic receptor family.

A new family of neuronal survival factors comprised of glial cell line-derived neurotrophic factor (GDNF) and neurturin has recently been described (Kotzbauer, P. T., Lampe, P. A., Heuckeroth, R. O., Golden, J. P., Creedon, D. J., Johnson, E. M., Jr., and Milbrandt, J. (1997) Nature 384, 467-470). These molecules, which are related to transforming growth factor-beta, are important in embryogenesis and in the survival of distinct neuronal populations. These molecules signal through a novel receptor system that includes the Ret receptor tyrosine kinase, a ligand (i.e. GDNF or neurturin), and an accessory glycosyl-phosphatidylinositol-linked molecule that is responsible for high affinity binding of the ligand. Two accessory molecules denoted GDNF family receptor 1 and 2 (GFRalpha-1 and GFRalpha-2) have been described that function in GDNF and neurturin signaling complexes. We have identified a novel co-receptor belonging to this family based on similarity to GFRalpha-1, which we have named GFRalpha-3. GFRalpha-3 displays 33% amino acid identity with GFRalpha-1 and 36% identity with GFRalpha-2. Despite the similarity of GFRalpha-3 to GFRalpha-1 and GFRalpha-2, it is unable to activate Ret in conjunction with GDNF, suggesting that there are likely additional undiscovered ligands and/or Ret-like receptors to be identified. GFRalpha-3 is anchored to the cell membrane by a phosphatidylinositol-specific phospholipase C-resistant glycosyl-phosphatidylinositol linkage. GFRalpha-3 is highly expressed by embryonic day 11 but is not appreciably expressed in the adult mouse. In situ hybridization analyses demonstrate that GFRalpha-3 is located in dorsal root ganglia and the superior cervical sympathetic ganglion. Comparison of the expression patterns of GFRalpha-3 and Ret suggests that these molecules could form a receptor pair and interact with GDNF family members to play unique roles in development.

Somatostatin modulates the function of Kupffer cells.

Recent studies have shown that somatostatin modulates lymphocyte and peritoneal macrophage function, but the effects of somatostatin on hepatic macrophages (Kupffer cells) are not clearly defined. In the present study, hepatic macrophages obtained from male rats were treated in vitro with somatostatin or octreotide and their effects on the release of nitrite, tumor necrosis factor (TNF), and hydrogen peroxide (H2O2) determined. At concentrations of 10(-14) M to 10(-12) M, or greater than 10(-10) M, somatostatin suppressed nitrite release by Kupffer cells. At concentrations of less than 10(-9) M or greater than 10(-9) M, octreotide inhibited nitrite release by Kupffer cells. Kupffer cells treated with 10(-10) M to 10(-14) M or greater than 10(-8) M of somatostatin released significantly less amounts of TNF than did the untreated controls. TNF release by Kupffer cells treated with 10(-15) M to 10(-5) M of octreotide was significantly inhibited as compared to that of untreated controls. Kupffer cells treated with 10(-14) M to 10(-11) M and 10(-9) M to 10(-8) M of somatostatin released more H2O2 than did the untreated controls. The amount of H2O2 released by noncirrhotic Kupffer cells treated with 10(-6) M or 10(-5) M of somatostatin was less than that of controls. These findings demonstrate that somatostatin and octreotide modulate the release of nitric oxide, TNF, H2O be Kupffer cells depending on the concentration of hormones used.

Glial cell line-derived neurotrophic factor signals through the RET receptor and activates mitogen-activated protein kinase.

Glial cell line-derived neurotrophic factor (GDNF), a member of the transforming growth factor-beta family of growth factors, was first identified by its ability to promote the survival of midbrain dopaminergic neurons in culture. We demonstrate that GDNF treatment of several neuroblastoma cell lines leads to dose-dependent tyrosine phosphorylation of the RET receptor and that other transforming growth factor-beta family members are not able to activate the RET receptor. GDNF treatment of neuroblastoma cells also results in increased transcription of an Elk luciferase reporter gene, suggesting that GDNF activates the mitogen-activated protein kinase signal transduction pathway.

Inhibition of the growth of a human nasopharyngeal carcinoma cell line by bFGF is mediated via FGFR-1.

The growth of CG-1 human nasopharyngeal carcinoma cell line and five of its randomly selected, single cell-derived subline cells is inhibited by bFGF in an autocrine and paracrine manner. In contrast, aFGF, which has a 55% homology in amino acid sequence with bFGF, stimulates cell growth. Basic FGF binds to specific cell surface high-affinity receptor sites with an apparent Kd of 105 pM. Of the two lines examined, the high-affinity binding sites for bFGF are calculated to be 1200 and 2600 per cell. The biological effect of bFGF is conveyed through its binding to the high-affinity receptor sites and the binding is dependent on the presence of cell surface heparin-like molecules, as treatment of cells with heparitinase or sodium chlorate abolishes high-affinity binding and growth inhibition. In contrast, similar treatment has no obvious effect on the growth-stimulatory effect of aFGF. Experimental results are also presented showing that the growth inhibition by bFGF is mediated through type I FGF receptors. These results suggest that bFGF and aFGF act via distinct receptor types to oppositely regulate the growth of CG-1 and subline cells.

Correlation of transformation from epithelial to mesenchymal-like morphology and endogenous bFGF levels in human nasopharyngeal carcinoma cells.

CG-1 human nasopharyngeal carcinoma cells in monolayer culture formed both cohesive, epithelial-like colonies and scattered, fibroblastic-like colonies in mixed proportions. In the presence of exogenously added bFGF (4 ng/ml), about 85% of the colonies formed were fibroblastic-like. CG-1 cells were capable of synthesizing and releasing bFGF, and, when compared by the immunological method, cells in fibroblastic-lke colonies were found to contain higher levels of endogenous bFGF than cells in the epithelial-like colonies. Furthermore, cells in the peripheral region of the epithelial-like colonies, which were fibroblastic-like in morphology, also appeared to contain higher levels of endogenous bFGF. In addition, in the presence of suramin, neutralizing antibody to bFGF, or neutralizing antibodies to bFGF and EGF, the number of cohesive colonies formed was greatly increased. Moreover, addition of the 2 M NaCl-eluted heparin-Sepharose fraction of the CG-1 cell-coditioned medium promoted the formation of dispersed colony in a dose-dependent manner. The results suggest that bFGF can regulate CG-1 cell phenotype in an autocrine manner.

Acidic FGF and EGF are involved in the autocrine growth stimulation of a human nasopharyngeal carcinoma cell line and sub-line cells.

The growth of a recently established human nasopharyngeal carcinoma cell line, CG-1, and 5 randomly selected, single-cell-derived sub-lines in serum-free medium and in fetal-bovine-serum(FBS)-containing medium was investigated. In basal medium supplemented with insulin, transferrin, fibronectin and high-density lipoprotein, cell growth was moderately stimulated by aFGF and EGF in a dose-dependent manner. In contrast, in medium containing as little as 0.5% FBS, most of the stimulatory effect of the aforementioned growth factors observed was masked. Western blotting analysis of the cell lysates and conditioned media showed that CG-1 and sub-line cells were all capable of synthesizing and releasing aFGF- and EGF-immunoreactive proteins. The amounts of these 2 growth factors synthesized and released appeared to vary among the parental cell line and sub-line cells. Moreover, the rate of basal proliferation of these cells appeared to be positively correlated with the amounts of aFGF- and EGF-immunoreactive proteins produced. Addition of the neutralizing antibodies to aFGF and EGF exerted a dose-dependent suppression on cell growth in medium containing 0.5% FBS. The results suggest a role of aFGF and EGF in autocrine growth stimulation of CG-1 and sub-line cells, and may explain the moderate response of these cells to exogenously added aFGF and EGF.

Tachykinins preferentially excite certain complex cells in the infragranular layers of feline striate cortex.

Microiontophoretically administered substance P (SP) affected the visually evoked responses (VER) and the spontaneous firing of 22 (14%) of the 152 neurons recorded from the striate cortex of anaesthetised cats. Enhancing effects were seen in 14 neurons and suppressant actions in 8 neurons. Most of the cells excited by SP were located in infragranular layers and had complex receptive fields; a few belonged to the movement-sensitive class or responded only weakly to visual stimulation. Of the neurons recorded in layer V, about 70% were excited by SP; the respective proportions were 8% in layer VI, and 2% in layer IV. Cells suppressed by SP had either simple or unimodal receptive fields including hypercomplex varieties; most of them were located in layer IVc. The effects of other tachykinins (neurokinin A, neurokinin B) and of the NK-3 receptor agonist Senktide tested in 36 cells were identical to those of SP with respect to types, and intracortical locations, of cells affected. During the enhancement induced by the tachykinins functional parameters of the neurons such as orientation and direction sensitivity were not substantially affected. It seems likely therefore that the effect of tachykinins in the primary visual cortex is not a shaping of receptive field properties, but rather a modulation of the general excitability of neurons projecting to subcortical centers, in particular to the midbrain and pons.

Mannose 6-phosphate receptor dependent secretion of lysosomal enzymes.

BHK and mouse L cells transfected with the cDNA for the human 46 kd mannose 6-phosphate receptor (MPR 46) secrete excessive amounts of newly synthesized mannose 6-phosphate containing polypeptides. The secretion is dependent on the amount, the recycling and the affinity for ligands of MPR 46. Incubation of transfected cells with antibodies blocking the binding site of MPR 46 reduces the secretion, and cotransfection with the cDNA for the human 300 kd mannose 6-phosphate (MPR 300) restores it to normal values. These results indicate that the two mannose 6-phosphate receptors compete for binding of newly synthesized ligands. In contrast to ligands bound to MPR 300, those bound to the MPR 46 are transported to and released at a site, e.g. early endosomes or plasma membrane, from where they can exit into the medium. Since antibodies blocking the binding site of MPR 46 reduce secretion also in non-transfected BHK and mouse L cells, at least part of the basal secretion of M6P-containing polypeptides is mediated by the endogenous MPR 46.

Epidemiologic characteristics and multiple risk factors of stomach cancer in Taiwan.

Both descriptive and analytical studies were carried out to examine epidemiologic characteristics and multiple risk factors of stomach cancer in Taiwan. The age-adjusted mortality rate of stomach cancer has been decreasing since the early 1970s for both males and females. The male-to-female ratio of the disease has remained around 2:1 in the past three decades. Comparison of the incidence of stomach cancer among Chinese in different countries showed a much lower incidence among Chinese in the USA than those in southeastern Asia. A hospital-based matched case-control study carried out in Taipei metropolitan areas showed a positive association of stomach cancer with blood type A, chronic gastric diseases, cigarette smoking, alcohol drinking, green tea drinking as well as consumption of salted meat, cured meat, smoked food, fried food and fermented beans. There was also a significant negative association between the disease and the consumption of milk.

Epidemiologic characteristics and multiple risk factors of lung cancer in Taiwan.

The specific aim of this study was to examine epidemiologic characteristics and multiple risk factors of lung cancer in Taiwan. The age-adjusted mortality from lung cancer has been increasing since the early 1950s with a constant male-to-female ratio of around 2.0. International comparison of cumulative mortality from lung cancer showed a much lower male-to-female ratio in Chinese than in other populations. Significantly high mortality from lung cancer was observed in highly urbanized cities and the endemic area of chronic arsenicism in Taiwan. Significant associations of active and passive cigarette smoking with epidermoid carcinoma, small cell carcinoma and adenocarcinoma of the lung were observed in a hospital-based case-control study carried out in Taipei metropolitan areas. Alcohol drinking, coffee drinking and various types of indoor air pollution were not related to lung cancer after the cigarette smoking habit was adjusted through a multiple logistic regression analysis.