Hemodynamic parameters and reproductive outcome after intracytoplasmic sperm injection and fresh embryo transfer in patients undergoing oocyte retrieval with general anesthesia using fentanyl,remifentanil or alfentanil – A randomized clinical trial

ObjectiveAnesthesia for assisted reproductive technology is very important to provide less stressful and painful environment for patients, with minimal side effects on oocytes. In the present study, we aimed to evaluate hemodynamic parameters, recovery time and intracytoplasmic sperm injection (ICSI) outcome among patients underwent anesthesia with fentanyl, remifentanil or alfentanil.Material and methodsThis randomized double-blinded clinical trial was conducted in patients undergoing anesthesia for transvaginal ultrasound guided oocyte retrieval (TUGOR). Patients were randomly allocated to alfentanil (A; 15 μg/kg), fentanyl (F; 1.5 μg/kg) or remifentanil (R; 1.5 μg/kg) groups.ResultsThree hundred forty patients were assessed for eligibility and randomized for transvaginal oocyte retrieval following general anesthesia and 105 were lost to follow up. No statistically significant differences were noted among groups regarding basic characteristics. Although, time to respond to verbal command was significantly different among groups (A: 1.99 ± 1.64, F: 2.56 ± 1.72, R: 1.78 ± 1.34, P = 0.014). There were no significant differences among groups with respect to the first and second postoperative pain intensity, patient satisfaction, pre-induction and post-induction systolic and diastolic blood pressure (BP). Terminal systolic (A: 101.61 ± 9.15, F: 105.29 ± 12.61, R: 102 ± 12.91, P = 0.01) and diastolic (A: 59.97 ± 9, F: 65.63 ± 9.13, R: 63.69 ± 11.01, P = 0.003) BP was significantly different among groups. The fertilization rate was significantly different among groups (A: 51.6%, F: 54.4%, R: 62.2%, P = 0.018). Implantation rate, biochemical and clinical pregnancy rate was similar among groups.ConclusionsThe results of present study demonstrated that all three opioids have the same efficiency, in regards to patient satisfaction and pregnancy outcome. However, Anesthesia with alfentanil compared with fentanyl and remifentanil, seems to be inferior for TUGOR due to higher effect on fertilization rate and less hemodynamic stability.Registration numberIRCT201410258677N4.     

Cloning,Expression, and Purification of Hyperthermophile α-Amylase from Pyrococcus woesei

Objectives

In an attempt α-amylase gene from Pyrococcus woesei was amplified and cloned into a pTYB2 vector to generate the recombinant plasmid pTY- α-amylase.

Methods

Escherichia coli BL21 used as a host and protein expression was applied using IPTG. SDS-PAGE assay demonstrated the 100 kDa protein. Amylolytic activity of proteins produced by transformed E. coli cells was detected by zymography, and the rate of active α-amylase with and without the intein tag in both soluble conditions and as inclusion bodies solubilized by 4M urea were measured.

Results

Amylolytic activity of ∼185,000 U/L of bacterial culture was observed from the soluble form of the protein using this system.

Conclusion

These results indicate that this expression system was appropriate for the production of thermostable α-amylase.     

Prognostic Values of Fluctuations in Serum Levels of Alanine Transaminase in Inactive Carrier State of HBV Infection

Background:

Current guidelines introduce periodic monitoring of serum alanine transaminase (ALT) as the first-line modality in follow-up patients, with a hepatitis B virus (HBV) inactive carrier state.

Objectives:

This study aimed to determine the incidence rate and patterns of ALT fluctuations and prognostic values for the development of chronic HBV e antigen (HBeAg)-negative hepatitis B (CHB), HBV surface antigen (HBsAg) seroclearance, and liver-related complications.

Patients and Methods:

Treatment-naïve patients with a chronic HBV infection, HBeAg(-)/HBeAb(+), normal ALT levels, and HBV DNA < 2000 IU/mL, were followed-up every 6-12 months by assessing serum ALT levels. Serum HBV DNA was measured in cases of elevated ALT levels.

Results:

A total of 399 patients were followed-up for 8.9 years; ALT > upper limit of normal (ULN, i.e. 40 IU/L) was detected in 103 (25.8%) patients, with an annual incidence rate of 2.9%. ALT elevation was associated with; male gender, age, and higher serum ALT levels at study entry. Among the cases of ALT elevations, 16 (15.5%) patients had ALT levels > 2 × ULN. There were 38 (36.9%) patients who had ALT levels that remained > ULN over six months, and 21 (20.4%) patients experienced at least two episodes of ALT elevations. In 15 (14.6%) patients, elevated ALT levels were associated with increased HBV replication (i.e. HBV DNA > 2 000 IU/mL) and these were considered as CHB. However, elevation of ALT levels, even in the absence of HBV replication, increased the risk for the development of CHB up to 8-fold in prospective follow-ups. HBsAg seroclearance, cirrhosis, and hepatocellular carcinoma were detected in 43 (10.8%), 4 (1%), and 1 (0.25%) patients, respectively.

Conclusions:

Fluctuations in serum ALT levels may change the prognosis of a HBV inactive carrier state.     

GAL3 receptor KO mice exhibit an anxiety-like phenotype

The neuropeptide galanin (GAL) is widely distributed in the central and peripheral nervous systems. It is a modulator of various physiological and pathological processes, and it mediates its effects via three G protein-coupled receptors (GAL_1–3 receptors). A role for GAL as a modulator of mood and anxiety was suggested, because GAL and its receptors are highly expressed in limbic brain structures of rodents. In recent years, numerous studies of animal models have suggested an involvement of GAL and GAL₁ and GAL₂ receptors in anxiety- and depression-related behavior. However, to date, there is sparse literature implicating GAL₃ receptors in behavioral functions. Therefore, we studied the behavior of GAL₃ receptor-deficient (GAL₃-KO) mice to elucidate whether GAL₃ receptors are involved in mediating behavior-associated actions of GAL. The GAL₃-KO mouse line exhibited normal breeding and physical development. In addition to behavioral tests, phenotypic characterization included analysis of hematology, amino acid profiles, metabolism, and sudomotor function. In contrast to WT littermates, male GAL₃-KO mice exhibited an anxiety-like phenotype in the elevated plus maze, open field, and light/dark box tests, and they were less socially affiliated than WT animals to a stranger mouse in a social interaction test. In conclusion, our data suggest involvement of GAL₃ receptors in GAL-mediated effects on mood, anxiety, and behavior, making it a possible target for alternative treatment strategies for mood disorders.Thirty years ago, Tatemoto et al. (1) isolated the neuropeptide galanin (GAL), a 29-aa (30-aa in humans) peptide, from porcine intestine. The peptide is highly conserved throughout evolution and found in many other species. GAL is widely distributed in the CNS and peripheral nervous system, and it has a variety of biological and physiological functions, ranging from energy homeostasis, reproduction, and feeding to cognition and learning (2). In the murine brain, GAL mRNA is extensively expressed in the hypothalamic and brainstem areas. The highest expression levels were observed in the preoptic, periventricular, and dorsomedial hypothalamic nuclei; bed nucleus of the stria terminalis (BNST); medial and lateral amygdala; locus coeruleus; and nucleus of the solitary tract (3). Furthermore, GAL coexists with the serotonin and norepinephrine systems in the rodent brain and acts as an inhibitory neuromodulator of norepinephrine, serotonin, dopamine, glutamate, and acetylcholine function (4). The expression pattern and neuromodulatory functions of GAL suggest a role for this neuropeptide in mood disorders like anxiety and depression. Accordingly, administration of GAL via the intracerebroventricular (i.c.v.) route or into the dopaminergic ventral tegmental area induced depression-like behavior in the rat forced swim test (FST) (5, 6). Several studies in GAL-overexpressing transgenic mice reported an increased depression-like behavior in the FST (7, 8) but found no differences in anxiety-related behavior under baseline conditions (7, 9). Holmes et al. (9) suggested, however, that GAL might have protective effects during periods of elevated stress, because GAL overexpression counteracted anxiogenic effects evoked by noradrenergic stimulation.To date, three GAL receptors (GAL_1–3 receptors) have been identified, and they are all members of the G protein-coupled receptor superfamily. The GAL receptor subtypes have substantial differences in their functional coupling and signaling activities, contributing to the diverse effects of GAL (2). In the CNS, GAL₁ and GAL₂ receptors are detected in the BNST, amygdala, hippocampus, hypothalamus, dorsal raphe nucleus, locus coeruleus, dorsal root ganglia, and thalamus. GAL₁ receptor is additionally expressed in the brainstem (medulla oblongata and lateral parabrachial nucleus) and in the dorsal horn of the spinal cord, and GAL₂ receptor expression is further found in the cerebral cortex, cerebellum, and spinal cord. Expression of GAL₃ receptor in the CNS is more limited, with mRNA being preferably detected in the hypothalamus (10, 11). This differential localization of the three GAL receptors in the brain, as determined by in situ hybridization, suggests that different functions of GAL might be mediated by individual receptor subtypes. Evidence from animal models indicates that all three GAL receptor subtypes are involved in functional processes related to anxiety and depression. Stimulation of the GAL₁ receptor with selective ligands results in a depression-like phenotype (12), whereas KO of this receptor in mice elicits increased anxiety-like behavior in the elevated plus maze (EPM) test but not in the light/dark (L/D) exploration, emergence, or open field (OF) tests (13). Consistent with an antidepressant-like effect of GAL₂ receptor signaling (14), Lu et al. (15) observed a depressive-like phenotype in GAL₂ receptor-deficient (GAL₂-KO) mice but found no GAL₂ receptor-mediated effects on anxiety-related behavior. GAL₃ receptor stimulation was suggested to induce a depression-like profile because decreased immobility and increased swimming in the rat FST were observed after treatment with the nonpeptidergic GAL₃ receptor-selective antagonist 1-phenyl-3-[[3-(trifluoromethyl)phenyl]imino]-1H-indol-2-one (SNAP 37889). Furthermore, SNAP 37889 induced anxiolytic-like behavior in the social interaction test (SIT) (16). In fact, GAL₃ receptor-selective antagonists like SNAP 37889 or SNAP 398299 and others (16, 17) have been and are still being developed to treat depressive disorders and/or anxiety. However, to our knowledge, any involvement of GAL₃ receptors in anxiety- or depression-related behavior was not previously verified in GAL₃ receptor-deficient (GAL₃-KO) animals. Therefore, we investigated behavior in a novel GAL₃-KO mouse line to elucidate whether the GAL₃ receptor is involved in mediating behavior-associated actions of GAL. Furthermore, for phenotypic characterization, analysis of GAL₃-KO animals included hematology, metabolism, and sudomotor function measurements.     

Neuropeptide Y and peptide YY protect from weight loss caused by Bacille Calmette–Guérin in mice

Background and Purpose

Immune challenge of mice with Bacille Calmette–Guérin (BCG) has been reported to cause transient weight loss and a behavioural sickness response. Although BCG-induced depression involves the kynurenine pathway, weight loss occurs independently of this factor. Because neuropeptide Y (NPY) and peptide YY (PYY) are involved in the regulation of food intake, we hypothesized that they play a role in the BCG-induced weight loss.

Experimental Approach

Male wild-type, PYY knockout (PYY−/−), NPY knockout (NPY−/−) and NPY−/−;PYY−/− double knockout mice were injected with vehicle or BCG (approximately 10⁸ colony-forming units per mouse), and their weight, locomotion, exploration and ingestion were recorded for 2 weeks post-treatment.

Key Results

Deletion of PYY and NPY aggravated the BCG-induced loss of body weight, which was most pronounced in NPY−/−;PYY−/− mice (maximum loss: 15%). The weight loss in NPY−/−;PYY−/− mice did not normalize during the 2 week observation period. BCG suppressed the circadian pattern of locomotion, exploration and food intake. However, these changes took a different time course than the prolonged weight loss caused by BCG in NPY−/−;PYY−/− mice. The effect of BCG to increase circulating IL-6 (measured 16 days post-treatment) remained unaltered by knockout of PYY, NPY or NPY plus PYY.

Conclusions and Implications

These data show that NPY and PYY are both required to protect from the action of BCG-evoked immune challenge to cause prolonged weight loss and disturb energy balance. The findings attest to an important role of NPY and PYY in orchestrating homeostatic reactions to infection and immune stimulation.     

The homeostatic role of neuropeptide Y in immune function and its impact on mood and behaviour

Neuropeptide Y (NPY), one of the most abundant peptides in the nervous system, exerts its effects via five receptor types, termed Y1, Y2, Y4, Y5 and Y6. NPY's pleiotropic functions comprise the regulation of brain activity, mood, stress coping, ingestion, digestion, metabolism, vascular and immune function. Nerve‐derived NPY directly affects immune cells while NPY also acts as a paracrine and autocrine immune mediator, because immune cells themselves are capable of producing and releasing NPY. NPY is able to induce immune activation or suppression, depending on a myriad of factors such as the Y receptors activated and cell types involved. There is an intricate relationship between psychological stress, mood disorders and the immune system. While stress represents a risk factor for the development of mood disorders, it exhibits diverse actions on the immune system as well. Conversely, inflammation is regarded as an internal stressor and is increasingly recognized to contribute to the pathogenesis of mood and metabolic disorders. Intriguingly, the cerebral NPY system has been found to protect against distinct disturbances in response to immune challenge, attenuating the sickness response and preventing the development of depression. Thus, NPY plays an important homeostatic role in balancing disturbances of physiological systems caused by peripheral immune challenge. This implication is particularly evident in the brain in which NPY counteracts the negative impact of immune challenge on mood, emotional processing and stress resilience. NPY thus acts as a unique signalling molecule in the interaction of the immune system with the brain in health and disease.