Immune responses to liver membrane antigens in patients with cystic fibrosis and liver disease

Biliary obstruction by viscid mucus, although important, may not be the only factor for the development of liver disease in some patients with cystic fibrosis. In the present study the relationship between immune responses to liver antigens and the presence of liver damage was investigated using the leucocyte migration test and lymphocyte cytotoxicity to isolated rabbit hepatocytes. Inhibition of leucocyte migration by purified liver-specific lipoprotein, derived from hepatocyte plasma membrane, was found in 9 of 11 children with liver disease, but in only 5 of 14 with cystic fibrosis and no overt liver disease (P < 0.025). Lymphocyte toxicity to isolated rabbit hepatocytes was significantly increased in 10 of 13 children with liver disease, but in only 6 of 29 children without liver disease (P < 0.001). Experiments using lymphocyte subpopulations showed that the cytotoxicity was mediated by a non-T-cell population and could be blocked with liver-specific lipoprotein in 7 out of 10 cases, suggesting that the reaction in these patients was specifically directed against liver-specific lipoprotein. The study suggests that sensitisation against liver membrane antigens, whether arising primarily or secondarily in some way to other hepatic lesions, may contribute to the progression of liver damage in cystic fibrosis.     

Results of 1-year screening of donors in The Netherlands for human T− lymphotropic virus (HTLV) type I: significance of Western blot patterns for confirmation of HTLV infection

BACKGROUND: Between January 1993 and January 1994, Dutch blood banks screened approximately 674,000 volunteer donors for the presence of antibodies to human T-lymphotropic virus type I (HTLV-I). STUDY DESIGN AND METHODS: Confirmatory testing was performed on samples from 870 different anti-HTLV-I-reactive donors (0.13% of the total tested). RESULTS: According to the authors' Western blot (WB) interpretation criteria, 15 (0.002%) donors tested HTLV-I-positive in the WB; 201 tested negative, and 654 (75% of donors reacting on enzyme-linked immunosorbent assay) tested indeterminate. Fresh samples from 234 of 870 anti-HTLV-reactive donors were tested for HTLV-I and type II (HTLV- II) DNA by polymerase chain reaction: all 15 WB-positive donors tested positive for HTLV-I DNA; 206 WB-indeterminate and 13 WB-negative donors tested negative for HTLV-I and -II DNA. Application of the manufacturer's (World Health Organization-based) guidelines for WB interpretation would have resulted in the misclassification of 48 (23%) of 206 polymerase chain reaction-negative donors as HTLV-I/II-positive. Risk factors were present in 14 of 15 HTLV-I-infected donors: 8 had a partner from an HTLV-I-endemic area, 4 were from HTLV-I-endemic countries, and 2 had received blood transfusions. CONCLUSION: HTLV-I and -II infection is rare among Dutch blood donors. HTLV screening will prevent few cases of HTLV-related disease, but it will prevent a further spread of the virus by transfusion. In a low-risk population, conservative guidelines for WB interpretation unnecessarily generate an excess of false-positive results.     

Validation of a new immunoblot assay (LiaTek HIV III) for confirmation of human immunodeficiency virus infection

BACKGROUND: Used as a supplemental assay, new anti-human immunodeficiency virus (HIV) immunoblots, employing recombinant and synthetic antigens, appeared to resolve the majority of samples with false-reactive Western blot results. Would it be possible to completely replace the Western blot by an immunoblot for confirmation and exclusion of HIV infection? STUDY DESIGN AND METHODS: The sensitivity of the new LiaTek HIV III immunoblot assay (Organon Teknika, Turnhout, Belgium) was tested on 416 Western-blot positive samples (386 HIV-1, 22 HIV-2, 1 HIV-1/2, and 7 HIV-O) and on 45 HIV-1 seroconversion samples. The specificity was tested on 146 samples from noninfected donors with false-positive results on a HIV screening test. RESULTS: All Western- blot-positive samples tested positive in the immunoblot (sensitivity: 100%). The immunoblot could not discriminate between HIV-1 and HIV-2 infection in 22 of 416 (5%) samples. The LiaTek assay showed reactivity in 28 of 45 seroconversion samples, whereas the Western blot reacted in 30 of 45 seroconversion samples. With false-positive donor samples, the immunoblot was indeterminate in 10 of 146 samples (specificity: 93%), and the Western blot was indeterminate in 44 of 146 samples (specificity: 70%). CONCLUSION: Like the Western blot, the immunoblot runs the risk of missing samples that are reactive by enzyme immunoassay during the early stage of HIV infection. Nevertheless, considering its superior specificity on false-positive donor samples, it appears that the immunoblot offers a cost-effective alternative to the Western blot assay for confirmation and exclusion of HIV infection.     

Immunotherapy‐responsive allodynia due to distal acquired demyelinating symmetric (DADS) neuropathy

Introduction: Distal acquired demyelinating symmetric (DADS) neuropathy is a distal variant of chronic inflammatory demyelinating polyradiculoneuropathy. It is characterized by chronic distal symmetric sensory or sensorimotor deficits. Sensory ataxia is a common clinical presentation. Nerve conduction studies typically show markedly prolonged distal motor latencies. Methods: We report 2 patients with chronic progressive generalized pain and fatigue, with normal neurological examinations except for allodynia. Results: Nerve conduction studies were typical of DADS neuropathy. Monoclonal protein studies were negative. Cerebrospinal fluid protein levels were elevated. Sural nerve biopsies revealed segmental demyelination and remyelination. One biopsy had marked endoneurial and epineurial lymphocytic infiltration. Immunomodulatory therapy alleviated the pain and fatigue and markedly improved distal motor latencies in both patients. Conclusions: DADS neuropathy can present with pain and a normal neurological examination apart from allodynia. Nerve conduction studies are necessary for diagnosis. These patients respond to immunotherapy better than typical DADS neuropathy patients with sensory ataxia. Muscle Nerve 54 : 973–977, 2016     

Use of complementary and alternative medicine by patients with cancer: a cross-sectional study at different points of cancer care

Complementary and alternative medicine (CAM) is widely used by cancer patients. In order to learn more on the usage of CAM, its reasons and motifs as well as sources of information along the trajectory of treatment, we decided to evaluate the prevalence and predictors for the use of CAM by cancer patients while being under active treatment with chemo- or radiotherapy or in aftercare. We distributed a standardized questionnaire among patients attending a department of radio-oncology, an ambulance for oncology and offices of general practitioners (GPs). Five hundred and six patients took part. Most attributed cancer to stress and trauma (23.7 and 16.4 %) or genes (20.8 %). Forty-four percentage reported knowing a physician with competence in CAM, and in all settings, most patients named the GP. Fifty-one percentage admitted using CAM, 35 % informed the oncologist about using CAM, 56 % informed the GP, and 26 % did not inform any physician. Most often used CAM was vitamin D (17 %) and selenium (16 %). Most important goals were to strengthen the immune system (59 %) and become active (52 %). Most patients were satisfied with the CAM methods they used. Yet, with some methods, dissatisfaction was up to 30 %. The GP has an important function concerning CAM in oncology as most patients believe the GP to have best knowledge in CAM. In order to integrate complementary medicine into evidence-based medicine, physicians should be trained on how to communicate on CAM with the patient and with each other. Explaining cancer and cancer therapies in a way lay persons are able to understand may be helpful. Physicians should actively address patients’ needs of involvement not only in decision making, but also actively in the therapy.     

Reduced lysosomal clearance of autophagosomes promotes survival and colonization of Helicobacter pylori

Evasion of autophagy is key for intracellular survival of bacteria in host cells, but its involvement in persistent infection by Helicobacter pylori, a bacterium identified to invade gastric epithelial cells, remains obscure. The aim of this study was to functionally characterize the role of autophagy in H. pylori infection. Autophagy was assayed in H. pylori‐infected human gastric epithelium and the functional role of autophagy was determined via genetic or pharmacological ablation of autophagy in mouse and cell line models of H. pylori infection. Here, we showed that H. pylori inhibited lysosomal function and thereby promoted the accumulation of autophagosomes in gastric epithelial cells. Importantly, inhibiting autophagosome formation by pharmacological inhibitors or genetic ablation of BECN1 or ATG5 reduced H. pylori intracellular survival, whereas inhibition of lysosomal functions exerted an opposite effect. Further experiments demonstrated that H. pylori inhibited lysosomal acidification and the retrograde trafficking of mannose‐6‐phosphate receptors, both of which are known to positively regulate lysosomal function. We conclude that H. pylori subverts autophagy into a pro‐survival mechanism through inhibition of lysosomal clearance of autophagosomes. Disruption of autophagosome formation offers a novel strategy to reduce H. pylori colonization in human stomachs. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

上颌骨缝牵张的组织学变化

目的:观察缝牵张过程中环上颌骨缝的组织反应和组织再生机制。方法:实验于2005-03/09在解放军第四军医大学口腔医学院颌面外科实验室完成。15周龄杂种犬12只,随机数字法分为实验组8只,对照组4只。实验组安置口外自制牵引支架,自鼻腭孔引出牵引钩,橡皮圈连接牵引支架和牵引钩,向前持续弹性牵引,牵引力约600g。牵引1周和4周时,各处死实验组犬4只和对照组犬2只,对前颌缝,腭横缝,颧颌缝,颧颞缝区组织作大体观察和组织学观察。结果:实验犬12只均进入结果分析。①骨缝区成骨主要发生在侧带与骨缘之间,缝牵张初期骨缘外侧出现囊状分离带,成骨细胞与成纤维细胞大量增殖,新骨与结缔组织纤维排列方向与牵引力方向基本平行。②中央带纤维较致密,在牵引过程中,纤维排列方式未发生明显变化,而起到了屏障作用,阻止了缝骨性融合的发生。③牵张早期,缝区组织同时可见Ⅰ和Ⅲ型胶原,随后成骨细胞在Ⅲ型胶原基质的基础上分泌沉积Ⅰ型胶原而成骨,最终Ⅲ型胶原降解,Ⅰ型胶原完全代替Ⅲ型胶原成为骨基质的主要成份。缝区组织天狼猩红染色未见Ⅱ型胶原。结论:缝牵张成骨是以膜内成骨方式为主,无明显的软骨内成骨现象。中央带具有屏障作用,阻止牵张过程中骨缝发生骨性融合。     

自体血清培养人骨髓间充质干细胞及其向神经细胞分化

目的:目前多采用胎牛血清培养骨髓间充质干细胞,实验应用自体血清培养骨髓间充质干细胞并诱导其向神细胞分化,拟验证其可行性及并分析其可能的机制。方法:实验于2005-10/2006-08在同济医学院附属同济医院神经外科实验室完成。培养骨髓间充质干细胞所用骨髓来源为本院需行骨髓穿刺的健康成人(共3例,年龄26～44岁),本人及家属知情同意。梯度离心法分离成人骨髓间充质干细胞,并利用自体血清进行体外培养扩增。主要观察指标:采用相差显微镜观察细胞形态变化;透射电镜观察骨髓间充质干细胞的超微结构;流式细胞术鉴定细胞表面标记物;3～5代细胞利用生长因子和N2添加剂进行诱导分化,采用RT-PCR检测Netin和神经元特异性烯醇化酶mRNA在诱导前后的表达。结果:①细胞形态变化:利用自体血清培养的骨髓间充质干细胞主要呈长或宽扁的梭形,在体外传代扩增速度较快。②超微结构:电镜下细胞表面可见细胞核较大,不规则,核质比较小,胞质中有丰富的细胞器,可见缝隙连接。③细胞表面标记物鉴定:多数细胞为CD44、CD105阳性,CD34、CD45阴性。④Netin和神经元特异性烯醇化酶mRNA的表达:诱导后部分细胞呈神经元样改变,Nestin mRNA表达水平于3d达高峰,随后减少,神经元特异性烯醇化酶mRNA于5～7d达高峰,并可持续数日。结论:应用自体血清可成功培养骨髓间充质干细胞,并在体外条件下定向诱导分化为神经元样细胞。