The Descending Projection of Serotoninergic , Fibers in the Raphe Nucleus of Brainstem and Its, Adjacent Areas in the Rat——A Double Labeled Study of HRP and Immunohistochemistry

Rats(Sprague Dawley,200-300g)wereoperated under anesthesia with nembuta1.Afterthe laminectomy,20-25%aqueous solution ofHRP(Sigma 6)was injected unilatera11y intothe spinal cord at cervical or lumbar leve1.Theanimals were allowed to survive for 36-48hours and transcardically perfused with 0.9%NaCl,3.5%paraforma1dehyde,and 10%sucr-ose in 0.1mol/L PB.The brain was removed andimmersed in 10%sucrose at 4℃.The brainstemwas cut serially at 40μm on freezing microtomy     

Role of PKC in the effects of α1-adrenergic stimulation on Ca2+ transients, contraction and Ca2+ current in guinea-pig ventricular myocytes

 The effects of α₁-adrenoceptor stimulation on intracellular Ca²⁺ transients, contractility and L-type Ca²⁺ current (I _Ca,L) were studied in single cells isolated from ventricles of guinea-pig hearts. The aim of our study was to elucidate the mechanisms of the positive inotropic effect of α₁-adrenergic stimulation by focussing on the role of protein kinase C (PKC). Phenylephrine, an α₁-adrenergic agonist, at concentrations of 50–100 μM elicited a biphasic inotropic response: a transient negative inotropic response (22.9±6.0% of control) followed by a sustained positive inotropic response (61.0±8.4%, mean±SE, n=12). The Ca²⁺ transient decreased by 10.2±3.9% during the negative inotropic phase, while it increased by 67.7±10% (n=12) during the positive inotropic phase. These effects were inhibited by prazosin (1 μM), a α₁-adrenergic antagonist. Phenylephrine increased the I _Ca,L by 60.8±21% (n=5) during the positive inotropic phase. To determine whether activation of PKC is responsible for the increases in Ca²⁺ transients, contractile amplitude and I _Ca,L during α₁-adrenoceptor stimulation, we tested the effects of 4β-phorbol 12-myristate 13-acetate (PMA), a PKC activator, and of bisindolylmaleimide I (GF109203X) and staurosporine, both of which are PKC inhibitors. PMA mimicked phenylephrine’s effects on Ca²⁺ transients, contractile amplitude and I _Ca,L. PMA (100 nM) increased the Ca²⁺ transient, contractile amplitude and I _Ca,L by 131±17%, 137±25% (n=8), and 81.1±26% (n=5), respectively. Prior exposure to GF109203X (1 μM) or staurosporine (10 nM) prevented the phenylephrine-induced increases in Ca²⁺ transients, contractile amplitude and I _Ca,L. Our study suggests that during α₁-adrenoceptor stimulation increase in I _Ca,L via PKC causes an increase in Ca²⁺ transients and thereby in the contractile force of the ventricular myocytes. Received: 16 July 1998 / Received after revision and accepted: 20 October 1998     

Effect of Andrographis paniculata as an Adjuvant in Combined Chemo-Radio and Whole Body Hyperthermia Treatment—A Preliminary Study

Modulation of immune responses to alleviate disease has been of interest for a long time. Intraperitoneal administration of Andrographis paniculata extract along with whole body hyperthermia (WBH) was found to enhance the total WBC count in cyclophosphamide (CTX) and radiation treated animals when compared to untreated control animals. Maximum inhibition in the solid tumor development was observed when the CTX and radiation exposed animals were treated with extract in combination with whole-body hyperthermia. Similarly myeloperoxidase activity in tumor tissue from CTX and radiation-treated animals was also significantly inhibited when they were administered with Andrographis paniculata extract along with whole body hyperthermia. Moreover the production of cytokines such as IL-2 and GM-CSF, which was reduced after combined CTX and radiation treatment was significantly increased by the simultaneous treatment of extract and whole body hyperthermia. The elevated level of serum Tumor Necrosis Factor (TNF-α) level, after CTX and radiation treatment was also lowered significantly after the administration of extract and simultaneous exposure of whole-body hyperthermia with respect to untreated tumor-bearing animals.     

Expression of Genes and Proteins of the Sarcoplasmic Reticulum Са2+-Transport Systems in Cardiomyocytes in Concomitant Coronary Heart Disease and Type 2 Diabetes Mellitus

Bulletin of Experimental Biology and Medicine - We compared the expression of Са2+-ATPase (SERCA2a), calsequestrin (CASQ2), ryanodine receptors (RyR2) proteins and their genes (ATP2A2,...     

Chaperone activity of α-crystallin in relation to the sarcoplasmic reticulum Ca2+ pump of skeletal muscles in stress and adaptation

α-Crystallin, an endogenous low-molecular-weight protein with chaperone activity, exerted protective effects on membrane systems of Ca²⁺ transport into the sarcoplasmic reticulum of skeletal muscles. Protective action of α-crystallin depended on the body state. This effect was not observed in the control and after adaptation to stress, while after stress, especially against the background of adaptation, α-crystallin increased the rate of Ca²⁺ transport into the sarcoplasmic reticulum and thermal resistance of Ca²⁺ pump. The mechanisms of α-crystallin activation during stress are discussed. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 9, pp. 279–282, September, 1999     

Does Post-Warming Extended Culture Duration Affect the Clinical and Obstetric Outcomes of Patients of Advanced Maternal Age? A Single-Center Study

BackgroundThe single vitrified-warmed blastocyst transfer (SVBT) cycle has been increasingly utilized for assisted reproductive technology. Women of advanced maternal age (AMA) comprise a significant portion of patients who have undergone ‘freeze-all’ cycles. This study investigated the association between the post-warming extended culture duration and pregnancy outcomes in patients of AMA.MethodsThis retrospective cohort study analyzed the outcomes of 697 SVBT cycles between January 2016 and December 2017. The cycles were divided into 3 groups based on the age of the female partners: group I: < 35 years (n = 407), group II: 35–37 years (n = 176); and group III, 38–40 years (n = 114). Data are shown as the mean ± standard error of the mean. Data were analyzed using one-way ANOVA followed by Duncan’s multiple range test. Statistical significance was set at P < 0.001.ResultsThe blastocyst rate, clinical pregnancy rate, and live birth rate (LBR) was significantly lower in the AMA groups. However, there were no significant differences in LBR in the transfer between the AMA and younger groups according to blastocyst morphology and post-warming extended culture duration.ConclusionPost-warming extended culture of blastocysts is not harmful to patients of AMA. It could be a useful parameter in clinical counseling and decision making for fertility treatments.     

How does the apolipoprotein E genotype modulate the brain in aging and in Alzheimer's disease? A review of neuroimaging studies

The epsilon 4 allele of apolipoprotein E is a risk factor for Alzheimer's disease, but also a modulator of its clinical picture. In this paper, recent research in neuroimaging of aging and Alzheimer's disease in relation to apolipoprotein E is reviewed, emphasizing the advances but also the controversies. Further, the possible clinicopathological implications of these findings are discussed.     

How will Internet Use Affect the Patient? A Review of Computer Network and Closed Internet-based System Studies and the Implications in Understanding How the Use of the Internet Affects Patient Populations

The widespread use of the Internet by patients is transforming the delivery of health information. Little research has been done, however, to assess the relationship between patients' use of online health resources and self efficacy, behavior or health status. To understand these effects and create a national research agenda, professionals should establish theoretically based studies. This article provides an overview of studies using computer networks and Internet-based closed systems in which a specific population has access to online health tools similar to those available on the Internet. These studies provide a microcosm of the effects Internet use may have on a patient's health-related behaviors. Three areas of proposed research will be explored: content research; process research; and outcomes research.     

Voltage-dependent Na+ and Ca2+ currents in human pancreatic islet β-cells: evidence for roles in the generation of action potentials and insulin secretion

We describe three voltage-dependent inward currents in human pancreatic -cells. First, a rapidly inactivating Na⁺ current, blocked by tetrodotoxin (TTX) is seen upon brief depolarization to or beyond –40 mV. Second, a transient, low-voltage-activated (LVA), amiloride-blockable Ca²⁺ current is seen upon depolarization to or beyond –55 mV; it inactivates within less than 1s of sustained depolarization to –40 mV. Third, a more sustained, high-voltage-activated (HVA) Ca²⁺ current, which shows variable sensitivity to dihydropyridines is seen upon depolarization to or beyond –40 mV, and thereafter slowly inactivates over a time course of many seconds. Our pharmacological evidence suggests that all three currents contribute to action potential initiation and upstroke when the background membrane potential (V _m) is equal or negative to –45 to –40 mV, a situation often induced by glucose concentrations (5–6 mM) in the range of those seen post-prandially. Consistent with this, TTX drastically reduces both transient and sustained insulin secretion in the presence of 5–6 mM glucose, but has little effect in 10 mM glucose, at which concentration cells rapidly depolarize to –35 mV, a V _m sufficient to rapidly inactivate Na⁺ and LVA Ca²⁺ currents.     

Correlations between alterations in length-dependent Ca2+ activation of cardiac myofilaments and the end-systolic pressure–volume relation

We have tested the hypothesis that alterations in length dependent activation (LDA) of cardiac myofilaments represent an important regulatory mechanism affecting the Frank-Starling mechanism as determined by the slope (E(es)) of the relation between left ventricular (LV) volume and end-systolic pressure. We employed a transgenic (TG) mouse model in which the cardiac isoform of TnI (cTnI) has been completely replaced with slow skeletal TnI (ssTnI), the embryonic/neonatal isoform in the heart. Compared to non-transgenic (NTG) controls, myofilaments from TG-ssTnI hearts demonstrate an increase in Ca(2+) sensitivity and a substantially blunted LDA that is unaffected by PKA-dependent phosphorylation. We measured in situ LV pressure and volume relations during basal conditions and isoproterenol (ISO) stimulation. In the basal state in TG-ssTnI hearts there was significant increase in end-systolic pressure and slight decrease in heart rate. ISO stimulation resulted in a significant increase in heart rate, ejection fraction, maximum dP/dt, preload-recruitable stroke work, maximum dP/dt versus end diastolic volume and cardiac output in both groups. During basal conditions there was no difference in the E(es) relation between NTG and TG-ssTnI groups. However, during ISO stimulation the E(es) relation was significantly different between NTG and TG-ssTnI groups. Our study provides the first direct evidence that enhancement in differences in LDA between cardiac myofilaments from NTG and TG-ssTnI hearts induced by post-translational modifications of sarcomeric proteins are reflected in the in situ beating heart by a different change in E(es). Thus, changes in LDA should be considered in interpreting results from in situ experiments on inotropic effects associated with physiological and patho-physiological states of the heart.     

Time resolved kinetics of the guinea pig Na–Ca exchanger (NCX1) expressed in Xenopus oocytes: voltage and Ca2+ dependence of pre-steady-state current investigated by photolytic Ca2+concentration jumps

Kinetic properties of the Na–Ca exchanger (guinea pig NCX1) expressed in Xenopus oocytes were investigated by patch clamp techniques and photolytic Ca²⁺ concentration jumps. Current measured in oocyte membranes expressing NCX1 is almost indistinguishable from current measured in patches derived from cardiac myocytes. In the Ca–Ca exchange mode, a transient inward current is observed, whereas in the Na–Ca exchange mode, current either rises to a plateau, or at higher Ca²⁺ concentration jumps, an initial transient is followed by a plateau. No comparable current was observed in membrane patches not expressing NCX1, indicating that photolytic Ca²⁺ concentrations jumps activate Na–Ca exchange current. Electrical currents generated by NCX1 expressed in Xenopus oocytes are about four times larger than those obtained from cardiac myocyte membranes enabling current recording with smaller concentration jumps and/or higher time resolution. The apparent affinity for Ca²⁺ of nonstationary exchange currents (0.1 mM) is much lower than that of stationary currents (6 μM). Measurement of the Ca²⁺ dependence of the rising phase provides direct evidence that the association rate constant for Ca²⁺ is about 5 × 10⁸ M⁻¹ s⁻¹ and voltage independent. In both transport modes, the transient current decays with a voltage independent but Ca²⁺-dependent rate constant, which is about 9,000 s⁻¹ at saturating Ca²⁺ concentrations. The voltage independence of this relaxation is maintained for Ca²⁺ concentrations far below saturation. In the Ca–Ca exchange mode, the amount of charge translocated after a concentration jump is independent of the magnitude of the jump but voltage dependent, increasing at negative voltages. The slope of the charge–voltage relation is independent of the Ca²⁺ concentration. Major conclusions are: (1) Photolytic Ca²⁺ concentration jumps generate current related to NCX1. (2) The dissociation constant for Ca²⁺ at the cytoplasmic transport binding site is about 0.1 mM. (3) The association rate constant of Ca²⁺ at the cytoplasmic transport sites is high (5 × 10⁻⁸ M⁻¹s⁻¹) and voltage independent. (4) The minimal five-state model (voltage independent binding reactions, one voltage independent conformational transition and one very fast voltage dependent conformational transition) used before to describe Ca²⁺ translocation at saturating Ca²⁺ concentrations is valid for Ca²⁺ concentrations far below saturation.     

Blockade of Ca2+‐activated K+ channels in T cells: an option for the treatment of multiple sclerosis?

Voltage- and Ca(2+)-dependent K(+) channels in the membrane of both T and B lymphocytes are important for the cellular immune response. In the current issue of the European Journal of Immunology, Reich et al. demonstrate that selective blockade of the intermediate-conductance Ca(2+)-activated K(+) channel (the IK channel encoded by the KCNN4 gene) prevents cytokine production in the spinal chord and ameliorates the development of EAE caused by injection of myelin oligodendrocyte glycoprotein (MOG)(35-55) in mice. These data renew the focus on the IK channel as a potential target for the development of new immune-suppressant drugs for the treatment of autoimmune diseases.     

Differential effects of a green tea-derived polyphenol (−)-epigallocatechin-3-gallate on the acidosis-induced decrease in the Ca2+ sensitivity of cardiac and skeletal muscle

(-)-Epigallocatechin-3-gallate (EGCg), a green tea-derived polyphenol, has received much attention as a protective agent against cardiovascular diseases. In this study, we determined its effects on the acidosis-induced change in the Ca(2+) sensitivity of myofilaments in myofibrils prepared from porcine ventricular myocardium and chicken pectoral muscle. EGCg (0.1 mM) significantly inhibited the decrease caused by lowering the pH from 7.0 to 6.0 in the Ca(2+) sensitivity of myofibrillar ATPase activity in cardiac muscle, but not in skeletal muscle. Studies on recombinant mouse cardiac troponin C (cTnC) and chicken fast skeletal troponin C (sTnC) using circular dichroism and intrinsic and extrinsic fluorescence spectroscopy showed that EGCg bound to cTnC with a dissociation constant of approximately 3-4 muM, but did not bind to sTnC. By presumably binding to the cTnC C-lobe, EGCg decreased Ca(2+) binding to cTnC and overcame the depressant effect of protons on the Ca(2+) sensitivity of the cardiac contractile response. To demonstrate isoform-specific effects of the action of EGCg, the pH sensitivity of the Ca(2+) response was examined in cardiac myofibrils in which endogenous cTnC was replaced with exogenous sTnC or cTnC and in skeletal myofibrils in which the endogenous sTn complex was replaced with whole cardiac Tn complex (cTn). The results suggest that the binding of EGCg to the cardiac isoform-specific TnC or Tn complex alters the effect of pH on myofilament Ca(2+) sensitivity in striated muscle.     

Does human papillomavirus play a role in the development of bladder transitional cell carcinoma? A comparison of PCR and immunohistochemical analysis

AIM: To investigate the role of human papillomavirus (HPV) in the development of bladder transitional cell carcinoma (TCC). METHODS: Seventy eight paraffin wax embedded TCC samples were tested for the presence of HPV by two methods. First, immunohistochemistry was carried out using a polyclonal antibody capable of detecting the capsid protein of all known papillomaviruses. The second method was a consensus GP5+/6+ primer mediated polymerase chain reaction (PCR) technique, with the products analysed by both agarose gel electrophoresis and an enzyme immunoassay using type specific oligonucleotide probes for 10 different mucosal genotypes. To exclude false negative results because of the poor quality of DNA extracted from paraffin wax embedded samples, the series was extended to include 20 further blocks for which the corresponding snap frozen unfixed tissue was available. RESULTS: The two methods produced contrasting results, with 47 of the 78 samples positive for HPV antigen and none positive for HPV DNA. HPV DNA was not detected in the 20 additional paraffin wax embedded TCCs or in the 20 paired unfixed samples. In contrast, HPV DNA was amplified by PCR from all six of the paraffin wax embedded cervical carcinoma and anogenital wart control samples. CONCLUSION: The disparity between the two sets of results is probably caused by false positives resulting from the non-specificity of the polyclonal antibody used for immunohistochemistry. These results suggest that HPV is unlikely to play an aetiological role in the development of bladder TCC.