Surveillance of antibiotic resistance in invasive isolates of Neisseria meningitidis in Australia 1994-1999

A total of 1434 strains of Neisseria meningitidis isolated from cases of invasive meningococcal disease (IMD) in Australia between 1994 and 1999 were examined by standard methods for susceptibility to antibiotics used for treatment and prophylaxis. The proportion of isolates fully susceptible to penicillin decreased from 45% in 1994 to 26% in 1999 (P<0.001). All the other isolates were less sensitive to penicillin except for two meningococci with a penicillin MIC of 1 mg/l. The geometric mean penicillin MIC increased from 0.045 to 0.065 mg/l from 1994 to 1999. There was no significant difference in the geometric mean penicillin MICs of serogroup B and serogroup C meningococci. Penicillin susceptibility was significantly associated with a poorer outcome. Isolates from survivors of IMD had a higher geometric mean penicillin MIC (0.06 mg/l) than those from fatal cases (0.048 mg/l) (P< 0.001). This suggests that factors other than the decrease in susceptibility to penicillin observed were more relevant to outcome in IMD. All isolates were fully susceptible to ceftriaxone. Rifampicin resistance was infrequent (eight isolates in 6 years) and sporadic. A single isolate had decreased quinolone susceptibility. Despite the significant shift in susceptibility to penicillin recorded, this group of antibiotics remains a suitable treatment for IMD in Australia.     

Recurrent mosaic MTOR c.5930C > T (p.Thr1977Ile) variant causing megalencephaly,asymmetric polymicrogyria,and cutaneous pigmentary mosaicism: Case report and review of the literature

Genetic alterations leading to overactivation of mammalian target of rapamycin (mTOR) signaling result in brain overgrowth syndromes such as focal cortical dysplasia (FCD) and megalencephaly. Megalencephaly with cutis tri‐color of the Blaschko‐linear type pigmentary mosaicism and intellectual disability is a rare neurodevelopmental disorder attributed to the recurrent mosaic c.5930C > T (p.Thr1977Ile) MTOR variant. This variant was previously reported at low to intermediate levels of mosaicism in the peripheral blood of three unrelated individuals with consistent clinical findings. We report a fourth case of a 3‐year‐old female presenting with megalencephaly, obstructive hydrocephalus due to cerebral aqueductal stenosis, asymmetric polymicrogyria, dysgenesis of the corpus callosum, hypotonia, developmental delay, and cutaneous pigmentary mosaicism. Oligonucleotide and SNP chromosomal microarray (CMA), karyotype, and trio whole exome sequencing (WES) in the peripheral blood, as well as a targeted gene variant panel from fibroblasts derived from hyperpigmented and non‐hyperpigmented skin did not detect any abnormalities in MTOR or other genes associated with brain overgrowth syndromes. Unlike the previously reported cases, the de novo c.5930C > T (p.Thr1977Ile) MTOR variant was detected at 32% mosaicism in our patient only after WES was performed on fibroblast‐derived DNA from the hyperpigmented skin. This case demonstrates the tissue variability in mosaic expression of the recurrent p.Thr1977Ile MTOR variant, emphasizes the need for skin biopsies in the genetic evaluation of patients with skin pigmentary mosaicism, and expands the clinical phenotype associated with this pathogenic MTOR variant.     

Retrospective multicentre study on mechanical and enzymatic preparation of fresh and cryopreserved testicular biopsies

BACKGROUND: Isolation of sperm suitable for ICSI from fresh or frozen-thawed testicular sperm extraction (TESE) can be facilitated by mechanical or enzymatic processing of the samples. METHODS: A retrospective multicentre study was initiated to compare these two approaches. Eleven German centres provided data on their TESE cycles performed during the period 1996/1997. Quality of retrieved sperm, fertilization rates of injected oocytes, embryo quality, resulting pregnancy rates and evolution of pregnancies were evaluated. RESULTS: The percentage of cycles with at least some motile sperm available for injection was higher after mechanical preparation. Independent of the preparation method, fertilization rates were higher for motile compared with immotile sperm or elongated spermatids in all groups and in general higher for cryopreserved versus fresh samples. Embryo quality was significantly better after injection of motile sperm for all treatments and in particular after enzymatic versus mechanical processing of biopsies. Pregnancy rates were identical for embryos derived from sperm prepared mechanically or enzymatically from fresh or cryopreserved testicular samples. The abortion rate (32/172, 18.6%) and the rate of multiple implantations (32/140, 22.9%) were not different from results reported in the literature for ICSI using ejaculated sperm. CONCLUSION: In this retrospective multicentre study, no unequivocal advantage of one over the other preparation method could be identified in 839 ICSI cycles using testicular sperm from 549 patients.     

Daily versus thrice-weekly interferon alfa-2b plus ribavirin for the treatment of chronic hepatitis C in HIV-infected persons: a multicenter randomized controlled trial

Among HIV-infected persons, chronic hepatitis C virus (HCV) infection causes substantial morbidity and mortality. However, few studies have evaluated the safety and efficacy of interferon alfa (IFN) and ribavirin (RBV) therapy in co-infected persons. Accordingly, a randomized, controlled, open-label, multicenter trial was conducted to establish the safety, tolerability, and efficacy of IFN alfa-2b 3 mIU daily plus RBV 800 mg/d compared with IFN alfa-2b 3 mIU thrice weekly (TIW) plus RBV 800 mg/d in HCV treatment-naive, HIV-infected subjects with compensated liver disease and stable HIV disease. The primary endpoint was sustained virologic response (SVR), defined as an undetectable HCV RNA level 24 weeks after discontinuation of HCV therapy. At study entry, subjects in both groups were similar with respect to age, gender, HCV genotype, and HIV disease status. Of 180 randomized subjects, 162 received at least 1 dose of study medication, constituting the modified intention-to-treat population. After 12 weeks of therapy, 122 (75%) had serum HCV RNA levels assessed; of these subjects, early virologic response (undetectable HCV RNA or >2 log10 decrease from baseline) was observed in 33 (42%) and 13 (16%) of subjects taking daily and TIW IFN, respectively (P < 0.001). SVR was observed in 15 (19.0%) and 7 (8.4%) of subjects taking daily and TIW IFN, respectively (P = 0.05). Adverse events were similar in both groups. However, while no deaths or opportunistic infections were observed, nearly 30% of subjects stopped treatment due to adverse events and 7 subjects experienced a serious adverse event. In conclusion, SVR was achieved in 19% of HIV/HCV coinfected subjects treated with daily IFN plus RBV, but the effectiveness of therapy was substantially diminished by relatively high rates of treatment-related toxicity.     

Potentiation of neurotransmitter release coincides with potentiation of phosphatidyl inositol turnover. A possible in vitro model for long term potentiation

Carbachol (CCh) a cholinergic agonist which hydrolyses phosphatidyl-inositol bisphosphate (PIP2) to produce the breakdown products inositol trisphosphate (IP3) and diacylglycerol (DAG) was tested for its ability to induce [3H]norepinephrine ([3H]NE) release and to accumulate [3H]inositol phosphate ([3H]IP) under normal and membrane depolarizing conditions. Our results suggest two major points: first, muscarinic acetylcholine receptor (mAChR) agonists and depolarizing agents (of which KCl is the most effective) act in concert to induce potentiation of PI turnover and potentiation of neurotransmitter release. The simultaneous presence of both a depolarizing agent and a receptor agonist is obligatory for eliciting potentiatory effect. Facilitation of release by muscarinic agonist and K+, added together, was 2 to 5-fold above additivity and the levels of [3H]IP accumulated were 3-5-fold above additivity by K+ and CCh. Enhancement of release and of [3H]IP formation is reversed by pirenzepine, a muscarinic (MI) specific antagonist, Kdiss = 0.4 and 0.8 microM, respectively. Second, synergy of IP accumulation in correlation with synergy of neurotransmitter release elicited by mAChR activation and membrane depolarization, suggests a possible role for phospholipase C (PLC) in the bifurcating control of neurotransmitter release and for the involvement of PLC and voltage sensitive channels in mediation of long-term potentiation (LTP).     

蝎毒抗癌多肽对CNE-2Z细胞生长周期移行和p53表达的影响

目的 :观察蝎毒抗癌多肽 (APBMV)对CNE 2Z细胞生长周期和p5 3表达的影响。方法 :采用流式细胞仪双标法。结果 :APBMV处理CNE 2Z细胞 48h后 ,进入S期的细胞明显减少 ,处于G1期和G2期的细胞明显增多 ,APBMV 16mg/L处理CNE 2Z细胞 48h后 ,G1期、S期和G2期细胞的百分比分别为 5 9.7%、32 .3 %和 7.9% ,空白对照组的百分比分别为 5 3 .3%、45 .2 %和 1.5 % ,二者比较差异有显著性 (P <0 .0 1或P <0 .0 0 1)。空白对照组CNE 2Z细胞P5 3蛋白 2 4h和 48h表达量处于较高水平 (2 7.2 3± 0 .93)和 (31.6 7± 1.75 ) ,经APBMV处理 2 4h或 48h后 ,P5 3蛋白表达量明显下降 ,与空白对照组相比差异有显著性 (P <0 .0 1)。结论 :APBMV能够阻止CNE 2Z细胞周期的移行 ,并抑制抑癌基因p5 3的表达     

东亚钳蝎蝎毒的指纹图谱

目的 :探索东亚钳蝎蝎毒 (scorpionofButhusmartensiivenom)的分析及鉴定方法 ,建立相关质量标准。方法 :采用高效液相色谱法 (highperformanceliquidchromatography ,HPLC)分析检测 8个蝎毒样品。结果 :蝎毒可分离出40多个组分 ,实验所使用的 8个省内所产蝎毒具有非常相似的图谱。蝎淋巴液和 3种蛇毒与蝎毒图谱有显著不同。同一样品 (二次冻干粉 )连续测定 5次进样量的精密度、重现性均达到有关规定。结论 :采用HPLC法获得了蝎毒的指纹图谱 ,并测定了可影响蝎毒质量的蝎淋巴液和可能用于掺伪的 3种蛇毒的图谱。本研究为蝎毒及其制剂的进一步开发应用提供了有价值的实验依据。     

癌组织中p16基因甲基化分析

目的探讨抑癌基因p16在胃癌组织中是否存在甲基化异常及其与胃癌发生发展的关系.方法对20例胃癌组织及相应正常胃粘膜组织应用甲基敏感酶(HpaII)和甲基非敏感酶(MspI)酶切,结合PCR扩增技术,对p16基因外显子1、外显子2的二核苷酸胞嘧啶特定序列5'-CCGG-3'位点甲基化进行检测.结果20例胃癌组织中,p16基因外显子1、2异常甲基化分别为5例(25%)和9例(45%),正常组织未发现甲基化异常;14例高甲基化标本中,中分化胃癌4例,低分化7例,高分化1例;有2例存在外显子1、2同时甲基化异常,二者均为低分化胃癌,进展期胃癌(Ⅲa、Ⅳ期各1例)中1例呈现泳动易位;外显子2甲基化异常多发生于晚期肿瘤患者(P<0.05).结论p16基因甲基化异常可能会造成基因功能丧失,从而失去对细胞增殖的负性调控作用,导致胃癌发生与进展;外显子2高甲基化与临床进展有关,可能为晚期事件.     

蝎毒抗癌多肽及其分离组分分析及鉴定方法

目的 :探讨蝎毒抗癌多肽 (antineoplasticpolypeptidefromButhusmartensiivenom ,APBMV)及其分离组分的分析及鉴定方法。方法 :采用SepharoseFF阳离子交换凝胶柱 (2 5cm× 5cm交换柱 ,流速 0 .8ml/min ,梯度洗脱 12 0 0min)层析法分离APBMV ,用高效毛细管电泳 (highperformancecapillaryelectrophoresis,HPCE)及高效液相色谱法 (highperformanceliquidchromatography ,HPLC)分析检测。结果 :用SepharoseFF阳离子交换凝胶层析可将APBMV分离为 2个峰。经HPCE、HPLC分析显示 ,APBMV主要成分为峰Ⅰ～Ⅳ ,其分离组分分别以峰Ⅰ或峰Ⅲ为主 ,其纯度分别为86 .35 %、6 9.5 7%。结论 :SepharoseFF用于进一步分离APBMV可望获得较高纯度的单一有效成分 ,HPCE及HPLC均可较好地显示蝎毒中小分子多肽的组成及相对含量 ,其结果基本一致     

Incidence of accidental awareness during general anaesthesia in obstetrics: a multicentre,prospective cohort study

General anaesthesia for obstetric surgery has distinct characteristics that may contribute towards a higher risk of accidental awareness during general anaesthesia. The primary aim of this study was to investigate the incidence, experience and psychological implications of unintended conscious awareness during general anaesthesia in obstetric patients. From May 2017 to August 2018, 3115 consenting patients receiving general anaesthesia for obstetric surgery in 72 hospitals in England were recruited to the study. Patients received three repetitions of standardised questioning over 30 days, with responses indicating memories during general anaesthesia that were verified using interviews and record interrogation. A total of 12 patients had certain/probable or possible awareness, an incidence of 1 in 256 (95%CI 149–500) for all obstetric surgery. The incidence was 1 in 212 (95%CI 122–417) for caesarean section surgery. Distressing experiences were reported by seven (58.3%) patients, paralysis by five (41.7%) and paralysis with pain by two (16.7%). Accidental awareness occurred during induction and emergence in nine (75%) of the patients who reported awareness. Factors associated with accidental awareness during general anaesthesia were: high BMI (25–30 kg.m^-2); low BMI (<18.5 kg.m^-2); out-of-hours surgery; and use of ketamine or thiopental for induction. Standardised psychological impact scores at 30 days were significantly higher in awareness patients (median (IQR [range]) 15 (2.7–52.0 [2–56]) than in patients without awareness 3 (1–9 [0–64]), p = 0.010. Four patients had a provisional diagnosis of post-traumatic stress disorder. We conclude that direct postoperative questioning reveals high rates of accidental awareness during general anaesthesia for obstetric surgery, which has implications for anaesthetic practice, consent and follow-up.