Diagnosis of Trichomoniasis in Male Patients on Performing Nested Polymerase Chain Reaction.

OBJECTIVE: Trichomoniasis is a parasitic infection that occurs with the settlement of Trichomonas vaginalis in female and male urinary and reproductive tracts. This infection is generally asymptomatic in males, and males are thought to be a carrier for the transmission of infection. In this study, our aim was to detect trichomoniasis using nested polymerase chain reaction among males who were referred to a hospital with suspected urinary tract infection. METHODS: Urine samples were collected from 138 male patients between 18 and 50 years of age who were referred with suspected urinary system infection to the Urology Outpatient Clinic at Malatya University Medical Center Malatya between December 2013 and May 2014. Direct microscopy, two different culture methods, and nested Polymerase chain reaction (PCR) were performed for the investigation of T. vaginalis in urine samples. RESULTS: Urinary tract infection was diagnosed in 47 of the 138 patients according to white and red blood cell counts in the urine samples. T. vaginalis infection was detected in 6.5% (9/138) of the suspected patients by nested PCR, while none of the samples tested positive by direct microscopy and culture examinations. Statistical significance was found between infection of the urinary tract and nested PCR positivity for T. vaginalis. CONCLUSIONS: According to our results, nested PCR is the most sensitive method for the detection of trichomoniasis in male patients. We strongly recommend using nested PCR for the differential diagnosis of urinary infections in males.

VIP down-regulates the inflammatory potential and promotes survival of dying (neural crest-derived) corneal endothelial cells ex vivo: necrosis to apoptosis switch and up-regulation of Bcl-2 and N-cadherin.

The neuropeptide vasoactive intestinal peptide (VIP) is anti-inflammatory and protective in the immune and nervous systems, respectively. This study demonstrated in corneal endothelial (CE) cells injured by severe oxidative stress (1.4 mM H(2)O(2)) in bovine corneal organ cultures that VIP pre-treatment (0, 10(-10), 10(-8), and 10(-6) M; 15 min), in a VIP concentration-dependent manner, switched the inflammation-causing necrosis to inflammation-neutral apoptosis (showing annexin V-binding, chromatin condensation, and DNA fragmentation) and upheld ATP levels in a VIP antagonist (SN)VIPhyb-sensitive manner, while up-regulated mRNA levels of the anti-apoptotic Bcl-2 and the differentiation marker N-cadherin in a kinase A inhibitor-sensitive manner. As a result, VIP, in a concentration-dependent and VIP antagonist-sensitive manners, promoted long-term CE cell survival. ATP levels, a determining factor in the choice of apoptosis versus necrosis, measured after VIP pre-treatment and 0.5 min post-H(2)O(2) were 39.6 +/- 3.3, 50.8 +/- 6.2, 60.1 +/- 4.8, and 53.6 +/- 5.3 pmoles/microg protein (mean +/- SEM), respectively (p < 0.05, anova). VIP treatment alone concentration-dependently increased levels of N-cadherin (Koh et al. 2008), the phosphorylated cAMP-responsive-element binding protein and Bcl-2, while 10(-8) M VIP, in a VIP antagonist (SN)VIPhyb-sensitive manner, increased ATP level by 38% (p < 0.02) and decreased glycogen level by 32% (p < 0.02). VPAC1 (not VPAC2) receptor was expressed in CE cells. Thus, CE cell VIP/VPAC1 signaling is both anti-inflammatory and protective in the corneal endothelium.

Cardiac inotropic and hypertrophic effects of Nigella sativa supplementation in rats.

The intrinsic cardiac responses to 2-months Nigella sativa (black seed) oral supplementation (800 mg/kg) in rats were investigated. The isolated hearts were perfused in a Langendorff preparation to demonstrate the effects of N. sativa on the baseline inotropic and chronotropic functions and the myocardial flow rate. N. sativa supplementation induced moderate global (homogenous) cardiac hypertrophy, evident by significant increases in left ventricular and whole heart weights as well as the relative heart weight/body weight ratio. The isolated perfused hearts of Nigella-treated rats showed enhanced levels of baseline peak tension, maximum rate of tension development, heart rate and myocardial flow rate. Moreover, there was a significant increase in the tension developed per gram left ventricular weight. The present study provides the first evidence of a physiological cardiac hypertrophy in rats induced by long term N. sativa supplementation.

Effects of Nigella sativa supplementation for one month on cardiac reserve in rats.

Nigella sativa (N. sativa) has a long history of use in folk medicine. In a current study performed in this laboratory, two-month dietary supplementation with N. sativa extract to normal rats has shown a homogenous cardiac hypertrophy and enhanced cardiac contractility at baseline conditions. In the present study, shorter (one-month) duration of oral N. sativa administration was adopted to detect possible earlier cardiac responses. In addition, in vitro cardiac stress by the beta adrenoceptor agonist isoproterenol was used to assess the intrinsic cardiac reserve mechanisms. The hearts of Nigella-treated rats developed a moderate but significant hypertrophy that was evident by an increase in the heart weight to body weight ratio. The observed Nigella-induced cardiac hypertrophy was associated with an increase in the baseline cardiac inotropic properties as well as the maximal peak tension generation upon progressive cardiac stress by isoproterenol infusion. The demonstrated selective enhancement of the inotropic reserve favours the physiological nature of Nigella-induced cardiac hypertrophy, similar to that provoked by exercise training.

Multiple cellular and subcellular actions of general anaesthetics on cultured molluscan neurones.

The pond snail Lymnaea stagnalis has been used as a model system to study the cellular and subcellular actions of general anaesthetics. Here we describe the actions of general anaesthetics mainly on cultured, identified neurones, maintained in isolation to prevent the actions of synaptic inputs upon them. Using the whole-cell patchclamp technique, we have found that application of clinical concentrations of inhalational anaesthetics (halothane and isoflurane) and barbiturates depresses whole-cell calcium currents and potassium currents in a concentration-dependent manner. After loading cultured neurones with the ratiofluorescent dye fura-2AM, we find that halothane raises intracellular calcium concentration in a concentration-dependent manner, both in the presence and absence of extracellular calcium. Thus anaesthetics have multiple cellular and subcellular actions, some of which we have described, but most of which are yet to be discovered.

Effects of general anaesthetics on cultured Lymnaea neurones.

In order to elucidate the mode of action of general anaesthetics we are using neurones of Lymnaea stagnalis as a model system. Neurones exhibit mainly two types of responses to anaesthetics delivered at clinical concentrations, i.e., either gradually going into quiescence or exhibiting paroxysmal depolarizing shifts (PDS). In order to determine whether these differences are due to intrinsic membrane properties or because of synaptic effects, cultured neurones are being used so that cells can be studied in isolation from any synaptic effects. Cells in culture retain their basic electrophysiological characteristics and behave in a similar manner to the applied anaesthetics as do whole brain preparations. Demonstration of PDS and quiescence in cultured neurones shows that these phenomena are due to membrane effects and not due to synaptic inputs. The effects of anaesthetics observed seem to be consistent with the suggestion that anaesthetics may influence the inward calcium current or other calcium-dependent currents.

Differential effects of general anaesthetics on identified molluscan neurones in situ and in culture.

1. The only unifying principle of general anaesthesia is that general anaesthetics interact with membrane components and no single cellular mechanism appears to explain their widespread effects in the central nervous system. 2. The gastropod mollusc, Lymnaea stagnalis, provides an excellent model system for studies on general anaesthetics because it has large, uniquely identifiable nerve cells. Several of these cells are interneurones with identified neurotransmitters and monosynaptic connections to other cells. 3. Recent work on Lymnaea neurones suggests that calcium currents are depressed by volatile general anaesthetics applied in the clinical range, whilst evidence from other preparations indicates that there is a rise in intracellular calcium concentration following application of these substances. 4. Identified Lymnaea neurones have different responses to applied anaesthetics, irrespective of the anaesthetic used. Following application of halothane, barbiturates and several other anaesthetic agents, some cells gradually become quiescent after a short period, whilst in others a series of paroxysmal depolarizing shifts occur prior to quiescence. 5. Cultured neurones of Lymnaea, Helisoma and related species retain their characteristic action potential types and neurotransmitter identity. Their responses to anaesthetics are similar to those in the intact brain. They may also form synapses in culture. Thus, they are a useful tool for studying the cellular and subcellular actions of general anaesthetics.