Video microscopy of intracellular pH in primary cultures of rabbit proximal and early distal tubules

The purpose of this study was to investigate intracytoplasmic pH (pH_i) regulation in primary cultures of proximal (PCT) and distal bright (DCTb) convoluted tubules. PCT and DCTb segments were microdissected from rabbit kidney cortex and cultured in a hormonally defined medium. The cultured epithelia were grown on semi-transparent permeable supports. The pH_i was determined by video microscopy and digital image processing using 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF) and measuring the ratio of BCECF fluorescence excited by two successive wavelengths (490 nm and 450 nm). Resting pH_i values, determined in bicarbonatefree medium (extracellular pH: 7.40), were 7.25±0.02 (n=23) and 7.17±0.04 (n=30) for cultured PCT and DCTb respecitively. After the acid-loading procedure, cultured proximal cells recovered their pH_i by means of the classic Na⁺/H⁺ antiporter, sensitive to amiloride and located in the apical membrane only. In cultured DCTb part of the pH_i recovery was mediated by a Na⁺/H⁺ exchange present in the basolateral side. Moreover, at physiological initial pH_i values, chloride removal from the apical solution caused the pH_i to increase in the presence of bicarbonate. In acidified cultured DCTb cells, a partial pH_i recovery was induced in sodium-free media by 15 mM HCO ₃ ^– in the presence of an outward chloride gradient. This pH_i change was completely abolished by 4,4-diisothiocyanostilbene 2,2-disulfonic acid (1 mM). These data suggest that DCTb cells possess in apical anion/base exchanger that resembles the Na⁺-independent Cl^–/HCO ₃ ^– exchanger.     

Education,training, and practice among nordic neuropsychologists. Results from a professional practices survey

Abstract

Objective: To investigate sociodemographic characteristics, clinical and academic training, work setting and salary, clinical activities, and salary and job satisfaction among practicing neuropsychologists in four Nordic countries. Methods: 890 neuropsychologists from Denmark, Finland, Norway, and Sweden participated in an internet-based survey between December 2013 and June 2015. Results: Three-fourths (76%) of the participants were women, with a mean age of 47 years (range 24–79). In the total sample, 11% earned a PhD and 42% were approved as specialists in neuropsychology (equivalent to board certification in the U.S.). Approximately 72% worked full-time, and only 1% were unemployed. Of the participants, 66% worked in a hospital setting, and 93% had conducted neuropsychological assessments during the last year. Attention deficit hyperactivity disorder, learning disability, and intellectual disability were the most common conditions seen by neuropsychologists. A mean income of 53,277 Euros was found. Neuropsychologists expressed greater job satisfaction than income satisfaction. Significant differences were found between the Nordic countries. Finnish neuropsychologists were younger and worked more hours every week. Fewer Swedish neuropsychologists had obtained specialist approval and fewer worked full-time in neuropsychology positions. Danish and Norwegian neuropsychologists earned more money than their Nordic colleagues. Conclusion: This is the first professional practice survey of Nordic neuropsychologists to provide information about sociodemographic characteristics and work setting factors. Despite the well-established guidelines for academic and clinical education, there are relevant differences between the Nordic countries. The results of the study offer guidance for refining the development of organized and highly functioning neuropsychological specialty practices in Nordic countries.     

Recovery,citizenship, and psychosocial rehabilitation: A dialog between Brazilian and American mental health care approaches

ABSTRACT

Over the past 30 years, the Brazilian psychiatric reform, based on the anti-asylum social movement and the psychosocial rehabilitation model of care, transformed radically the country’s system of mental health care. Even though tremendous gains were made, people with serious mental illness continue to have limited access to citizenship. In the United States the recovery movement has enhanced the central role of people with serious mental illness as an active and responsible agent in their recovery process, encouraging a more collaborative role for service users, but, still, there are challenges to citizenship. This article first identifies commonalities between these two movements, central to mental health advances in the two distinct cultures of Brazil and the United States. Then, though celebrating the cultural differences between the two societies, the authors consider the possibility of cross-fertilization and propose a set of suggestions that could help both countries to overcome the remaining barriers to social justice for those living with mental health issues. From the American recovery movement, the authors learn that society can and must make adaptations to allow people with mental health disorders to live as citizens regardless of the symptoms they might manifest. From Brazil, mental health workers can use solidarity as a tool to create inventive solutions for those with a mental health disorder to live as citizens, regardless of the symptoms they might manifest.     

Selective IgG subclass deficiency: quantification and clinical relevance

Each of the four human IgG subclasses exhibits a unique profile of effector functions relevant to the clearance and elimination of infecting microorganisms. The quantitative response within each IgG subclass varies with the nature of the antigen, its route of entry and, presumably, the form in which it is presented to the immune system. This results in antibody responses to certain antigens being predominantly or exclusively of a single IgG subclass. An inability to produce antibody of the optimally protective isotype can result in a selective immunodeficiency state. This is particularly apparent for responses to certain bacterial carbohydrate antigens that are normally of IgG2 isotype. A failure to produce the appropriate specific antibody response may result in recurrent upper and/or lower respiratory tract infection. Careful patient investigation can identify such deficiencies and suggest appropriate clinical management. In this review we outline the biology and clinical relevance of the IgG subclasses and summarize current rational treatment approaches.     

兔关节软骨细胞的琼脂糖凝胶培养及其力学性质的研究

目的对兔膝关节软骨细胞进行体外琼脂糖凝胶三维培养,并对其力学性能进行检测。方法将琼脂糖凝胶、细胞、血清和培养基在正确的比例下混合,形成软骨细胞凝胶块。取培养7d、14d和21d时的软骨细胞凝胶块分别对细胞外基质进行组织学观察和免疫组化染色,同时采用Instron 5544材料试验机检测其极限应力、极限应变、抗压模量和切线模量的力学性能变化。结果在琼脂糖凝胶中培养的关节软骨细胞具有典型的软骨细胞特性,能够正常合成细胞外基质,形成有一定弹性和抗压缩能力的软骨样凝胶块。而且随着培养时间增长,基质合成增加,培养21d时的极限应力(0.0226±0.006)N/mm、抗压模量(0.608±0.061)N/mm和切线模量(0.096±0.004)N/mm,比7d时的(0.0204±0.004)MPa,(0.558±0.036)N/mm,(0.029±0.002)N/mm和14d时的(0.0213±0.008)N/mm,(0.586±0.095)N/mm,(0.049±0.005)N/mm有明显增加,但极限应变却没有明显差异(P>0.05)。结论软骨细胞-琼脂糖凝胶块的力学性能与细胞外基质的合成直接相关。     

IL-1β作用于人牙周膜成纤维细胞后TRAF6蛋白表达的研究

目的通过研究IL-1β干预下,TRAF6在人牙周膜成纤维细胞中的蛋白表达,为细胞因子网络对细胞功能调控作用的研究提供新资料。方法原代培养人牙周膜成纤维细胞,并进行免疫学鉴定。取5～8代细胞以IL-1β梯度浓度干预,采用免疫组织化学方法检测TRAF6在人牙周膜成纤维细胞中的表达。结果TRAF6在正常的人牙周膜成纤维细胞中呈阴性表达,IL-1β干预后,TRAF6呈阳性表达,并且随IL-1β干预的浓度的升高而增强。结论IL-1β作用后,TRAF6在牙周膜成纤维细胞出现表达,且与IL-1β作用浓度相关。     

Thyroxine induces transition of red towards white muscle in cultured heart cells

Summary Thyroid hormones (TH) have previously been shown to alter the force and velocity of cardiac muscle contractions. To investigate the mechanism responsible for these alterations, excess amounts of thyroxine (T₄, 1M) were applied on rat heart cells grown in cell culture. We found the following biochemical alterations: a) 40% decrease in the myoglobin content within 2 days; b) 25% increase in the rate of Ca-uptake into sacroplasmic reticulum (SR) in myocytes following chemical skinning; and c) a two-fold increase in Na–K-ATPase activity measured by⁸⁶Rb-uptake. These changes support our hypothesis that TH induce the transition of slow-twitch (red) muscles towards the fast-twitch (white) muscle type. This may explain the changes in contractile activity known to occur under TH influence.     

伯氏疟原虫的体外培养和扫描电镜观察

采用蜡烛缸法比较大鼠血清、人脐带血清、成人B型血清、兔血清、小牛血清和加有次黄嘌呤的小牛血清对伯氏疟原虫体外生长的影响.并对培养前和培养12、20、28h的小牛血清培养物作扫描电镜观察。结果表明,所用血清均不能明显改善长期培养效果;对短期培养的效果,以大鼠血清和人脐带血清为优,次黄嘌呤对疟原虫体外生长有潜在的抑制作用。电镜观察表明,小牛血清能使大鼠红细胞发生凝集,红细胞和疟原虫表面所形成的表面粘附物能妨碍裂殖子入侵和疟原虫对代谢物的吸收和排泄。     

人轮状病毒持续感染细胞系的建立

引起细胞病变的人轮状病毒DS-1株在MA-104细胞建立了持续感染状态。持续感染细胞系无明显细胞病变、生长良好,能持续携带和产生较高滴度的人轮状病毒,且所产生病毒活性较高,与正常DS-1病毒相似,持续感染细胞系具有极强的抗VSV攻击能力,而其培养液上清中无高水平干扰素存在。人轮状病毒持续感染细胞系的建立对病毒感染机理的研究和减毒活疫苗研制有一定现实意义。