Molecular interactions of the plasma membrane calcium ATPase 2 at pre- and post-synaptic sites in rat cerebellum

The plasma membrane calcium extrusion mechanism, PMCA (plasma membrane calcium ATPase) isoform 2 is richly expressed in the brain and particularly the cerebellum. Whilst PMCA2 is known to interact with a variety of proteins to participate in important signalling events [Strehler EE, Filoteo AG, Penniston JT, Caride AJ (2007) Plasma-membrane Ca(2+) pumps: structural diversity as the basis for functional versatility. Biochem Soc Trans 35 (Pt 5):919–922], its molecular interactions in brain synapse tissue are not well understood. An initial proteomics screen and a biochemical fractionation approach identified PMCA2 and potential partners at both pre- and post-synaptic sites in synapse-enriched brain tissue from rat. Reciprocal immunoprecipitation and GST pull-down approaches confirmed that PMCA2 interacts with the post-synaptic proteins PSD95 and the NMDA glutamate receptor subunits NR1 and NR2a, via its C-terminal PDZ (PSD95/Dlg/ZO-1) binding domain. Since PSD95 is a well-known partner for the NMDA receptor this raises the exciting possibility that all three interactions occur within the same post-synaptic signalling complex. At the pre-synapse, where PMCA2 was present in the pre-synapse web, reciprocal immunoprecipitation and GST pull-down approaches identified the pre-synaptic membrane protein syntaxin-1A, a member of the SNARE complex, as a potential partner for PMCA2. Both PSD95–PMCA2 and syntaxin-1A–PMCA2 interactions were also detected in the molecular and granule cell layers of rat cerebellar sagittal slices by immunohistochemistry. These specific molecular interactions at cerebellar synapses may allow PMCA2 to closely control local calcium dynamics as part of pre- and post-synaptic signalling complexes.     

流式细胞术检测化疗前后卵巢癌细胞凋亡的临床意义

目的：探讨化疗前后卵巢癌细胞及外周血淋巴细胞凋亡的因子Ｆａｓ、Ａｐｏ２．７的含量及其临床意义。方法：从７５例化疗前后卵巢癌患者的腹水中提取富含癌细胞的悬液,用Ａｎｎｅｘｉｎ Ｖ－ＦＴＴＣ／ＰＩ双染法分辨凋亡的亡细胞及坏死细胞及坏死细胞,并以两种凋亡因子克隆抗体Ｆａｓ和Ａｐｏ２．７标记上述细胞,均用流式细胞仪检测。结果：化疗后卵巢癌细胞凋亡率显著高于化疗前的临床疗效好,反之疗效差,被Ｆａｓ及Ａｐｏ２     

Influence of phenobarbital pretreatment on thein vitro ring a reduction of Δ4-3-oxo-steroids in rat liver microsomes

Summary The metabolism of 4-androstene-3,17-dione has been studied in rat liver microsomes. Treatment of the animals with repeated doses of phenobarbitalin vivo caused an enhanced rate of formation of 5-androstane-3,17-dione and of polar metabolites from this steroid. On the other hand, no significant increase was found in the 5-reductase activity present in the soluble cytoplasm after phenobarbital administration to the rats. Carbon monoxide and oxidized cytochrome c abolished the formation of polar metabolites from 4-androstene-3,17-dione but did not inhibit the 5-reductase activity. The present findings indicate that the hydroxylation and 5-reduction reactions, although both stimulated by phenobarbital treatment of the animals, do not involve common enzyme components.Supported by grants from the Swedisch Cancer Society, Swedish Medical Research Council (Project no. 13X-2525) and from Caroline Andriette Nobel's foundations for experimental medical research.     

Pediatric poisoning from trimedoxime (TMB4) and atropine automatic injectors

OBJECTIVE: To describe the effects of combined trimedoxime (TMB4) and atropine poisoning from automatic injectors (AI) in children. STUDY DESIGN: Data was collected from two sources: calls to the Israel Poison Information Center (IPIC) during a 1-year period and a cohort of children who presented to pediatric emergency departments (EDs) after unintentional injection of an AI. Demographic data and data regarding the type of AI, site and time of injection, and the clinical manifestations were abstracted. RESULTS: Data were available for 142 patients. The median age was 8.5 years (range 1.25-18 years). The dose of atropine and TMB4 was higher than the recommended dose for age in 22 (15.5%) cases. There were few side effects attributable to atropine: dilated pupils (26.7%), dryness of mucous membranes (24.6%), and tachycardia (22.5%). Compared with children injected with an age-appropriate dose, children injected with an AI that contained a dose that exceeds the recommended one were more likely to be symptomatic ( P = .029). There were no side effects characteristic to oximes, and no specific medical intervention was required. CONCLUSIONS: Unintentional pediatric atropine and TMB4 injection, even an adult dose in a small child, does not cause significant side effects.     

Decreasing overuse of therapies in the treatment of bronchiolitis by incorporating evidence at the point of care

OBJECTIVE: To describe the effect of evidence-based point-of-care algorithms and rules, based on guideline recommendations, on the overuse of therapies for bronchiolitis.Study design Pre-postintervention for infants <1 year of age admitted with a first-time episode of bronchiolitis. Data collected for guideline-eligible patients discharged between January 15, 2002, and March 27, 2002, were compared with data collected for guideline-eligible patients discharged from the hospital with a diagnosis of bronchiolitis during the same time period in the first 5 years after the original guideline implementation (1997 to 2001). The primary outcome of interest was use of bronchodilator therapy. Secondary outcomes included use of guideline order sets, resource utilization, length of stay, and readmission. RESULTS: A total of 256 patients from 2002 were compared with 1272 historic patients. In 2002, the odds of receiving any bronchodilator, more than 1, more than 2, and more than 4 bronchodilators were all significantly less than predicted by the 1997 to 2001 year-to-year trend. The odds of receiving a nasopharyngeal wash for respiratory syncytial virus and a chest radiography (OR=0.680, CL=0.476, 0.973) were also significantly lower than what was predicted from use trends of previous years. CONCLUSIONS: Evidence-based point-of-care instruments can have a significant effect on unwarranted treatment variation.     

Clinical and cost implications of target joints in Canadian boys with severe hemophilia A

OBJECTIVE: To compare target joint-associated costs incurred by boys with severe hemophilia A 1 year before and 1 year after development of a target joint (pre-TJ, post-TJ). STUDY DESIGN: Resource utilization data were extracted retrospectively from medical and hemophilia clinic charts and patient diaries for 16 boys attending the Hospital for Sick Children (HSC)'s comprehensive care hemophilia program. Resources examined included drugs, medical care, hospitalization, laboratory tests, therapies, and transfusions received. All costs were figured using standard price lists and were discounted using an annual rate of 3%. RESULTS: Fifteen of the 16 boys developed at least one target joint, defined as three bleeds into any single joint within a consecutive 3-month period, at an average age of 54 months (range, 15-94 months), with ankles being most often affected, followed by elbows and knees (46% vs 28% and 23%, respectively). The total cost of treating a boy with on-demand Factor VIII (FVIII) increased by 119% after development of a target joint, from $20,091 (in 2002 Canadian dollars [$CDN]) in the year before to $43,890 in the year after target joint development. Factor VIII use accounted for 87% of the total cost in the year before target joint development and 93% in the year after. CONCLUSIONS: This study identified substantial increased costs of care associated with target joint development. This finding provides further support for more aggressive treatment aimed at reducing target joints-either more aggressive treatment of joint bleeds or institution of primary prophylactic therapy at an early age.     

Drivers and barriers to seeking treatment for erectile dysfunction: a comparison of six countries

OBJECTIVE: To identify predictors of treatment-seeking behaviour in men with erectile dysfunction (ED) and the predictors (correlates) of individual drivers and barriers to seeking treatment. Although the prevalence and epidemiology of ED have been reviewed, there is little information about the treatment-seeking behaviour of men with this disorder. SUBJECTS AND METHODS: Data from the Cross-National Survey on Male Health Issues conducted between March and September 2000 were assessed by multivariate analysis. A cohort of 32 644 men aged 20-75 years was recruited during visits to their physicians. The men completed a short screening questionnaire, covering their overall health, and prostate, urinary and erectile problems. Men identified as having ED completed a detailed follow-up questionnaire. Logistic regression methods were used to identify predictors of treatment-seeking behaviour, and individual drivers and barriers to seeking treatment. RESULTS: Most men with ED had not sought treatment. The analyses suggested that ED, in conjunction with a desire to have sex, was necessary for men to seek treatment. Men seeking treatment commonly identified themselves as self-motivated or that they were influenced by a spouse or sex partner. The youngest group (20-39 years) was least likely to seek treatment. Among those who did not seek treatment, younger men were likely to believe that their ED would resolve spontaneously, whereas older men resisted seeking treatment because they felt that ED was a natural part of ageing. CONCLUSIONS: The data from this survey of men using the healthcare system confirmed other population-based reports that a minority of men with ED seek treatment. Subset analyses showed that treatment-seeking behaviour tended to be driven primarily by the man or by his sex partner. Common barriers to seeking treatment included the belief that ED would resolve spontaneously (younger men) and that ED was a normal part of ageing (older men).     

Penile revascularization surgery for arteriogenic erectile dysfunction: the long-term efficacy rate calculated by survival analysis

OBJECTIVE: To determine the subjective and objective outcomes (by survival analysis) after penile revascularization surgery in patients with arteriogenic erectile dysfunction (ED), selected by established strict criteria. PATIENTS AND METHODS: The study included 51 patients diagnosed with arteriogenic ED caused by localised arterial lesions and who had microscopic penile revascularization surgery between January 1996 and March 2002. Before surgery, all patients had a full examination, including a medical and sexual history, laboratory testing, intracavernosal pharmacological tests, colour Doppler ultrasonography (CDU), pharmacodynamic infusion cavernosometry and cavernosography, and digital subtraction angiography (DSA). Penile revascularization surgery was indicated only in patients aged <50 years and with no history of diabetes mellitus, hypertension or hyperlipidaemia. When there were communicating branches between the dorsal and cavernosal arteries, Hauri's procedure was used; when there were none or there was no evidence for them on both CDU and DSA because of severe narrowing or obstruction in the proximal common penile artery, the Furlow-Fisher modification of the Virag V procedure (FFV5) was used. The patency of the neo-arterial blood flow was assessed by CDU and effective rates calculated using the Kaplan-Meier method. The efficacy rate was recalculated whenever there was a recurrence. When occlusion of the neo-arterial blood was confirmed by CDU the date of occlusion was set as that midway between the last examination showing patency of the donor vessel and the latest examination indicating the occlusion. The patency period was the number of days from surgery to the date of occlusion. RESULTS: Of the 51 patients, 26 had Hauri's and 23 the FFV5 procedure (median age 32 years, range 21-49); in two patients with a previous pelvic fracture surgery was not possible because of scar formation in the dorsal area at the base of the penis. The mean (sd) subjectively estimated efficacy rate was 85.9 (6.3)% after 3 and 67.5 (10.7)% after 5 years of follow-up. The duration at 75% efficacy was 41.0 (5.6) months. The objectively estimated efficacy rate was 84.9 (7.3)% at 3 and 65.5 (13.5)% after 5 years of follow-up. The duration at 75% patency was 42.4 (9.5) months, and at 50% was 60.6 (19.4) months. There was no significant difference in subjective outcome between the FFV5 and Hauri procedures (P = 0.38, log rank test) and none objective outcome after surgery (P = 0.19, log rank test). Thirteen of the 18 patients in the Hauri group had venous dilatation in the deep dorsal, obturator, prostatic and the internal iliac veins. There were operative complications in four patients (hyperaemia of the glans in two, and one each with haemorrhage from the anastomosis site and scar contracture). CONCLUSIONS: The long-term efficacy rates (by the Kaplan-Meier method) of the Hauri and FFV5 procedures were both acceptable. The selection criteria gave acceptable outcomes from both procedures. Penile revascularization surgery is a treatment suitable only for young men and therefore attention must be given not only to the long-term outcome but also to long-term adverse events.