The role of ABC-transporter gene polymorphisms in chemotherapy induced immunosuppression, a retrospective study in childhood acute lymphoblastic leukaemia

We examined the association of functional ABCB1 (MDR1) and ABCG2 (BCRP) polymorphisms with acute side effects of chemotherapy. Analyses were performed on clinical data from 138 patients treated with the ALL-BFM-95 protocol implying several substrates of these transporters. ABCB1 3435T>C, 2677G>T/A 1236C>T and ABCG2 421C>A genotypes were determined. A higher proportion of ABCB1 3435TT patients suffered excessive infectious complications than those harbouring at least one C allele (OR=2.5, p=0.03) during the whole half-year-long intensive phase of chemotherapy. Weaker associations were calculated when ABCB1 1236T-2677T-3435T haplotype homozygotes were tested against the remaining part of the population (OR=2.3, p=0.09). During the reinduction phase of therapy, the occurrence of severe leukocytopenia was similar among ABCB1 genotype groups. The frequency of any toxicities were not shown to differ according to the ABCG2 421C>A genotype. Our data suggest that the ABCB1 3435T>C genotype is associated with the infectious complications of the applied chemotherapy regimen.     

Adenine aminohydrolase from Leishmania donovani: unique enzyme in parasite purine metabolism

Adenine aminohydrolase (AAH) is an enzyme that is not present in mammalian cells and is found exclusively in Leishmania among the protozoan parasites that infect humans. AAH plays a paramount role in purine metabolism in this genus by steering 6-aminopurines into 6-oxypurines. Leishmania donovani AAH is 38 and 23% identical to Saccharomyces cerevisiae AAH and human adenosine deaminase enzymes, respectively, catalyzes adenine deamination to hypoxanthine with an apparent K(m) of 15.4 μM, and does not recognize adenosine as a substrate. Western blot analysis established that AAH is expressed in both life cycle stages of L. donovani, whereas subcellular fractionation and immunofluorescence studies confirmed that AAH is localized to the parasite cytosol. Deletion of the AAH locus in intact parasites established that AAH is not an essential gene and that Δaah cells are capable of salvaging the same range of purine nucleobases and nucleosides as wild type L. donovani. The Δaah null mutant was able to infect murine macrophages in vitro and in mice, although the parasite loads in both model systems were modestly reduced compared with wild type infections. The Δaah lesion was also introduced into a conditionally lethal Δhgprt/Δxprt mutant in which viability was dependent on pharmacologic ablation of AAH by 2'-deoxycoformycin. The Δaah/Δhgprt/Δxprt triple knock-out no longer required 2'-deoxycoformycin for growth and was avirulent in mice with no persistence after a 4-week infection. These genetic studies underscore the paramount importance of AAH to purine salvage by L. donovani.     

A Modified EPA Method 1623 that Uses Tangential Flow Hollow-fiber Ultrafiltration and Heat Dissociation Steps to Detect Waterborne Cryptosporidium and Giardia spp.

Cryptosporidium and Giardia species are two of the most prevalent protozoa that cause waterborne diarrheal disease outbreaks worldwide. To better characterize the prevalence of these pathogens, EPA Method 1623 was developed and used to monitor levels of these organisms in US drinking water supplies ¹². The method has three main parts; the first is the sample concentration in which at least 10 L of raw surface water is filtered. The organisms and trapped debris are then eluted from the filter and centrifuged to further concentrate the sample. The second part of the method uses an immunomagnetic separation procedure where the concentrated water sample is applied to immunomagnetic beads that specifically bind to the Cryptosporidium oocysts and Giardia cysts allowing for specific removal of the parasites from the concentrated debris. These (oo)cysts are then detached from the magnetic beads by an acid dissociation procedure. The final part of the method is the immunofluorescence staining and enumeration where (oo)cysts are applied to a slide, stained, and enumerated by microscopy.Method 1623 has four listed sample concentration systems to capture Cryptosporidium oocysts and Giardia cysts in water: Envirochek filters (Pall Corporation, Ann Arbor, MI), Envirochek HV filters (Pall Corporation), Filta-Max filters (IDEXX, Westbrook, MA), or Continuous Flow Centrifugation (Haemonetics, Braintree, MA). However, Cryptosporidium and Giardia (oo)cyst recoveries have varied greatly depending on the source water matrix and filters used^1,14. A new tangential flow hollow-fiber ultrafiltration (HFUF) system has recently been shown to be more efficient and more robust at recovering Cryptosporidium oocystsand Giardia cysts from various water matrices; moreover, it is less expensive than other capsule filter options and can concentrate multiple pathogens simultaneously^{1-3,5-8,10,11}. In addition, previous studies by Hill and colleagues demonstrated that the HFUF significantly improved Cryptosporidium oocysts recoveries when directly compared with the Envirochek HV filters⁴. Additional modifications to the current methods have also been reported to improve method performance. Replacing the acid dissociation procedure with heat dissociation was shown to be more effective at separating Cryptosporidium from the magnetic beads in some matrices^9,13 .This protocol describes a modified Method 1623 that uses the new HFUF filtration system with the heat dissociation step. The use of HFUF with this modified Method is a less expensive alternative to current EPA Method 1623 filtration options and provides more flexibility by allowing the concentration of multiple organisms.     

Recurrent Herpetic Stromal Keratitis in Mice,a Model for Studying Human HSK

Herpetic eye disease, termed herpetic stromal keratitis (HSK), is a potentially blinding infection of the cornea that results in over 300,000 clinical visits each year for treatment. Between 1 and 2 percent of those patients with clinical disease will experience loss of vision of the infected cornea. The vast majority of these cases are the result of reactivation of a latent infection by herpes simplex type I virus and not due to acute disease. Interestingly, the acute infection is the model most often used to study this disease. However, it was felt that a recurrent model of HSK would be more reflective of what occurs during clinical disease. The recurrent animal models for HSK have employed both rabbits and mice. The advantage of rabbits is that they experience reactivation from latency absent any known stimulus. That said, it is difficult to explore the role that many immunological factors play in recurrent HSK because the rabbit model does not have the immunological and genetic resources that the mouse has. We chose to use the mouse model for recurrent HSK because it has the advantage of there being many resources available and also we know when reactivation will occur because reactivation is induced by exposure to UV-B light. Thus far, this model has allowed those laboratories using it to define several immunological factors that are important to this disease. It has also allowed us to test both therapeutic and vaccine efficacy.     

皮肤软组织感染病原菌分布及耐药性分析

目的 调查皮肤软组织感染病原菌的种类及耐药性,为临床合理使用抗菌药物提供科学依据.方法 利用WHONET 5.6对2009年1月至2011年12月皮肤软组织感染患者脓液或分泌物细菌培养及药敏试验结果进行回顾性分析.结果 共分离菌株444株,金黄色葡萄球菌(SAU)分离率居第1位,171株占38.5％;表皮葡萄球菌居第2位,54株占12.2％;铜绿假单胞菌第3位,43株占9.7％;头孢哌酮/舒巴坦、哌拉西林/他唑巴坦、阿米卡星、亚胺培南和美罗培南对革兰阴性杆菌有较好的抗菌活性(耐药率≤3.3％);金黄色葡萄球菌对青霉素和红霉素的耐药率为93.6％和65.0％,对呋喃妥因、利奈唑胺、利福平、莫西沙星和左旋氧氟沙星耐药率为0％、0％、1.4％、2.2％和9.3％,未检出万古霉素耐药株;耐甲氧西林金黄色葡萄球菌(MRSA)检出率25.7％ (44/171);MRSA和甲氧西林敏感金黄色葡萄球菌(MSSA)对氨苄西林/舒巴坦、利福平、莫西沙星三种药物的敏感性差异有统计学意义.结论 引起皮肤软组织感染病原菌以阳性球菌尤其是金黄色葡萄球菌为主,临床上应尽量根据药敏试验结果选用抗生素,合理用药.     

灰仓鼠多房棘球蚴腹腔动物模型的建立

目的观察多房棘球蚴(Echinococcus muhilocularis E.m)在灰仓鼠腹腔内的生长发育规律,制作E.m灰仓鼠动物模型。方法 2000个E.m原头节/只剂量腹腔接种灰仓鼠,接种后10、15、18、22、39、60、80和100 d,测量包囊重量和抗体,计算包囊重与灰仓鼠体重的包囊系数,观察多房棘球蚴在灰仓鼠体内的发育规律。分别按100个/只、500个/只和2000个/只剂量腹腔接种灰仓鼠各10只,100 d后剖检比较不同感染剂量的包囊发育情况和感染率。结果 62只灰仓鼠中有59只感染了多房棘球蚴。18 d可观察到成熟原头节,感染率达到100%,15 d出现抗体阳性,60 d时包囊系数达到15%以上;100、500和2000三种剂量的感染率100%;结论建立的灰仓鼠腹腔多房棘球蚴动物模型具有感染率高,感染周期短,包囊生长发育和抗体滴度变化易观察,感染60 d可作为E.m动物模型成模观察期。包囊系数,抗体滴度和包囊发育变化可作为造模的主要参考指标。雌性灰仓鼠造模优于雄性。     

伤寒沙门菌动物模型研究进展

目前伤寒沙门菌引起的人类伤寒仍是一种严重危害人类健康的疾病,且近年来多重耐药伤寒沙门菌株频频出现,使伤寒的治疗更加棘手.由于该菌具有严格的宿主特异性,又缺乏理想的动物模型,其致病机制的研究、疫苗及药物的研发受制约.新近研究发现,免疫系统人源化小鼠模型和诱导型一氧化氮合酶基因敲除小鼠模型可用于伤寒沙门菌的体内实验.本文就其应用现状及缺憾作一综述.     

多药耐药菌肺感染患者应用改良后呼吸机对病室环境卫生学影响

目的:探讨应用改良后的呼吸机对病室环境卫生学的影响。方法:对10例产碳青霉烯酶肺炎克雷伯菌肺感染应用呼吸机的患者和40例肺部感染耐甲氧西林金黄色葡萄球菌(MRSA)使用呼吸机的患者进行分组试验,对病室环境卫生学监测结果进行统计分析。机器改良方法是呼吸机的呼气阀处连接一条螺纹管,将带菌空气引出,消毒后排放。结果:肺部感染产碳青霉烯酶肺炎克雷伯菌应用改良后呼吸机的一组在空气、床头桌、水池、操作中人员手的阳性率分别为20%、10%、20%、10%,较未改良的对照组为80%、70%、70%、50%有明显的下降。肺部感染MRSA使用有创改良呼吸机组空气、床头桌、水池、操作中人员手的阳性率分别为10%、10%、5%、10%均低于无创改良组的20%、20%、10%、20%。结论:此种方法有效地降低了环境的污染程度,降低了医源性医院感染的风险。加强洗手和隔离等卫生预防学措施可以有效的降低MRSA的医院感染率。