Neural localization of addicsin in mouse brain

Addicsin is a member of the prenylated Rab acceptor (PRA) 1 domain family and a murine homolog of the rat glutamate-transporter-associated protein 3-18 (GTRAP3-18). This protein is considered to function as a modulator of the neural glutamate transporter excitatory amino acid carrier 1 (EAAC1). However, its molecular functions remain largely unknown. Here, we examined the regional and cellular localization of addicsin in the central nervous system (CNS) by using a newly generated antibody specific for the protein. Distribution analysis by Western blot and immunohistochemistry demonstrated that the protein was widely distributed in various regions of the mature CNS, including the olfactory bulbs, cerebral cortex, amygdala, hippocampus CA1–3 fields, dentate gyrus, and cerebellum. Double immunofluorescence analysis revealed that addicsin was expressed in the somata of principal neurons in the CNS such as the pyramidal cells and gamma-aminobutyric acid (GABA)-ergic interneurons scattered in the hippocampal formation. Furthermore, the protein showed pre-synaptic localization in the stratum lucidum of the CA3 field of the hippocampal formation. Subcellular localization analysis of highly purified synaptic fractions prepared from mouse forebrain supported the cytoplasmic and pre-synaptic distribution of addicsin. These results suggest that addicsin has neural expression and may play crucial roles in the basic physiological functions of the mature CNS.     

Hermansky–Pudlak syndrome type 2: Aberrant pre‐mRNA splicing and mislocalization of granule proteins in neutrophils

Hermansky–Pudlak syndrome type 2 (HPS2) is a syndrome caused by mutations in the beta‐3A subunit of the adaptor protein (AP)‐3 complex (AP3B1 gene). We describe five unreported cases with four novel mutations, one of which caused aberrant pre‐mRNA splicing. A point mutation c.2702C>G in exon 23 of the AP3B1 gene caused deletion of 112 bp in the mRNA in two siblings. This mutation activates a cryptic donor splice site that overrules the wild‐type donor splice site of this exon. Three other novel mutations in AP3B1 were identified, that is, a nonsense mutation c.716G>A (p.Trp239Ter), a 1‐bp and a 4‐bp deletion c.177delA and c.1839_1842delTAGA, respectively, both causing frameshift and premature termination of translation. Mass spectrometry in four of these HPS2 patients demonstrated the (near) absence of all AP‐3 complex subunits. Immunoelectron microscopy on the neutrophils of two of these patients showed abnormal granule formation. We found clear mislocalization of myeloperoxidase in the neutrophils even though the content of this protein but not the activity seemed to be present at normal levels. In sum, HPS2 is the result of the absence of the entire AP‐3 complex, which results in severe neutropenia with a defect in granule formation as the major hematological finding.     

稳心颗粒联合美托洛尔治疗冠心病心律失常的疗效分析

目的 探讨稳心颗粒联合美托洛尔治疗冠心病心律失常的临床疗效。方法 选择2016年1月~2017年1月我院收治的100例冠心病心律失常患者作为研究对象,随机分为观察组和对照组,每组50例,对照组采用稳心颗粒进行治疗,观察组采用稳心颗粒联合美托洛尔进行治疗,观察两组患者的治疗效果与不良反应。结果 观察组的临床效果优于对照组的临床效果,差异有统计学意义（P<0.05）;观察组的室性早搏发生频率、ST段压低、ST段压低持续时间低于对照组,;观察组的临床不良反应要低于对照组的情况,差异均具有统计学意义（P<0.05）。结论 稳心颗粒联合美托洛尔治疗冠心病心律失常的效果较为良好,具有高效性和安全性,值得在临床中广泛的应用和推广。     

小儿柴桂退热颗粒治疗小儿风寒感冒的临床分析

目的 分析小儿柴桂退热颗粒治疗小儿风寒感冒的临床疗效。方法 将2016年3月~2017年3月在我院儿科门诊患风寒感冒患儿88例进行研究,随机分为两组,各44例,对照组以小儿氨酚黄那敏颗粒治疗,观察组以小儿柴桂退热颗粒治疗,比较两组用药后咳嗽、咳痰、咽痛的治疗效果。结果 观察组咳嗽、咳痰、咽痛消失时间均短于对照组,治疗总有效率高于对照组,均存在统计学意义（P＜0.05）。不良反应仅对照组1例发生轻微恶心,不良反应发生率为2.27%。结论 小儿柴桂退热颗粒治疗小儿风寒感冒效果明显,可促使患儿临床症状尽早缓解,有助于减轻患儿痛苦,值得推广。     

Neuroprotective effect of carnosine in the olfactory bulb after vanadium inhalation in a mouse model

Carnosine (β‐alanyl‐L‐histidine) is synthesized in the olfactory system, has antioxidant activity as a scavenger of free radicals and has been reported to have neuroprotective action in diseases which have been attributed to oxidative damage. In neurodegenerative disorders, such as Parkinson's and Alzheimer's diseases, impairment of olfactory function has been described. Vanadium derivatives are environmental pollutants, and its toxicity has been associated with oxidative stress. Vanadium toxicity on the olfactory bulb was reported previously. This study investigates the neuroprotective effect of carnosine on the olfactory bulb in a mice model of vanadium inhalation. Male mice were divided into four groups: vanadium pentoxide (V₂O₅) [0.02 mol/L] inhalation for one hour twice a week; V₂O₅ inhalation plus 1 mg/kg of carnosine administered daily; carnosine only, and the control group that inhaled saline. The olfactory function was evaluated using the odorant test. Animals were sacrificed four weeks after exposure. The olfactory bulbs were dissected and processed using the rapid Golgi method; cytological and ultrastructural analysis was performed and malondialdehyde (MDA) concentrations were measured. The results showed evidence of olfactory dysfunction caused by vanadium exposure and also an increase in MDA levels, loss of dendritic spines and necrotic neuronal death in the granule cells. But, in contrast, vanadium‐exposed mice treated with carnosine showed an increase in dendritic spines and a decrease in neuronal death and in MDA levels when compared with the group exposed to vanadium without carnosine. These results suggest that dendritic spine loss and ultrastructural alterations in the granule cells induced by vanadium are mediated by oxidative stress and that carnosine may modulate the neurotoxic vanadium action, improving the olfactory function.     

Mast cell activation markers for in vitro study

ABSTRACT

Mast cells (MCs) are well known for their role in allergic conditions. This cell can be activated by various types of secretagogues, ranging from a small chemical to a huge protein. Mast cell activation by secretagogues triggers the increase in intracellular calcium (iCa²⁺) concentration, granule trafficking, and exocytosis. Activated mast cells release their intra-granular pre-stored mediator or the newly synthesized mediator in the exocytosis process, in the form of degranulation or secretion. There are at least three types of exocytosis in mast cells, which are suggested to contribute to the release of different mediators, i.e.,, piecemeal, kiss-and-run, and compound exocytosis. The status of mast cells, i.e., activated or resting, is often determined by measuring the concentration of the released mediator such as histamine or β-hexosaminidase. This review summarizes several mast cell components that have been and are generally used as mast cell activation indicator, from the classical histamine and β-hexosaminidase measurement, to eicosanoid and granule trafficking observation. Basic principle of the component determination is also explained with their specified research application and purpose. The information will help to predict the experiment results with a certain study design.     

Calcium reduces mitochondrial membrane potential of cultured rat cerebellum granule cells under toxic action of glutamate

Glutamate-induced decrease in mitochondrial membrane potential of granule cell is prevented by cobalt ions and the noncompetitive selective antagonist of NMDA-receptors MK-801. Similar to glutamate, the calcium ionophore A23187 reduces this potential. Translated fromByulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 123, No. 4, pp. 378–380, April, 1997     

An electron microscopic and immunohistochemical study of Merkel cell carcinoma

Two cases of Merkel cell carcinoma (MCC) were examined by electron microscopy and immunohistochemistry. Histologically, tumor cells, which extended from the dermis into the subcutis, showed anastomosing bands with partial trabecular pattern. The ultrastructural study showed tumor cells in case 1 with numerous neurosecretory granules. The number of granules in case 2, however, was smaller compared with that in case 1. Perinuclear bundles of filaments were present in case 2, but few bundles were observed in case 1. By immunohistochemistry, cytokeratin (CK)-8, -18, -19, and -20 and epithelial membrane antigen were stained positively within tumor cells in both cases. It was interesting that staining patterns of chromogranin A and of neuron-specific enolase were different in the two cases. These data indicated that CK-20 is a useful marker for diagnosing MCC and that ultrastructural and immunohistochemical differences in both cases were the result of phenotypic variation.     

Expression of reg protein in rat regenerating islets and its co-localization with insulin in the Beta cell secretory granules

Summary Regenerating islets can be induced by the administration of poly(ADP-ribose) synthetase inhibitors to 90% depancreatized rats. In screening a regenerating isletderived cDNA library, we previously isolated a novel gene, reg (regenerating gene), which encodes a 165-amino acid protein with a 21-amino acid signal sequence. In the present study, we have examined the expression and localization of reg protein in the regenerating islets by immunocytochemical techniques using a monoclonal antibody against a recombinant rat reg protein of 144 amino acids without the signal sequence. Light microscopy examination showed strong immunoreactivity for reg protein in the regenerating islets of the rats at two weeks and two months after the 90% pancreatectomy, whereas reg protein was almost undetectable in normal rat islets or in the islets of the rats one year after the pancreatectomy. Almost all the reg protein-positive cells were stained for insulin. By applying the immunogold technique at the ultrastructural level, it was demonstrated that both reg protein and insulin occur in the central granular core of the regenerating Beta cell secretory granules. These results suggest that reg protein is synthesized in and secreted from the regenerating Beta cells and that its expression is closely associated with Beta-cell regeneration.