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Cerebrospinal fluid western Blot profiles in the evolution of HIV-1 pediatric encephalopathy. 脑脊液western Blot分析HIV-1儿童脑病的演变
Pub Date : 1998-01-01
S M Ruţă, R Mătuşa, C C Cernescu

In order to obtain information on neurologic AIDS, 54 white caucasian children infected by nosocomial route with a median age of 46.2 +/- 7 months were followed up prospectively for a median of 12 months with three months Denver tests neurologic evaluation and six months serologic investigations in CSF and sera. Paired CSF and serum samples, collected on the same day, from children with AIDS encephalopathy, were analysed for the permeability of the blood brain barrier (BBB) and for intrathecal production of anti HIV specific antibodies. A prospective follow-up and repeated comparison of WB profiles and the presence of anti V3 antibodies in CSF and sera was done, as well as an evaluation of the modification in the CSF antibody specificity (anti gag Western Blot scoring) with disease progression. An increased intrathecal synthesis of IgG was recorded in all subjects, in spite of an unaltered BBB permeability. No significant differences were recorded for the anti gag score in the serum samples, which was stable between 9.1-10.4. By contrast, the score for CSF samples decreases significantly with disease progression, from 8.7 in children without encephalopathy, to 6.5 in those with stationary disease and 3.6 in the progressive encephalopathy group. A strong correlation was found between the level of anti p24 antibodies determined by ELISA and the anti gag score quantified by WB for the same CSF samples. The p24 antigen was found to be positive only in 3 cases, even after immune complex dissociation. Anti V3 antibodies were not detected in CSF samples from patients with functional BBB. The decline in anti-gag antibody reactivity in CSF is an early indicator of disease progression, reflecting a severe course of neurological impairments. The absence of anti V3 antibodies in the CSF samples suggests that the PND of neurotropic strains mapped in distinct positions into the V3 loop. These results reflect the selection of antigenic escape mutants which evolve in the CNS, distinct from the blood lymphotropic isolates.

为了获得神经系统艾滋病的相关信息,我们对54例经医院途径感染的白人儿童进行了中位年龄46.2±7个月的前瞻性随访,随访时间中位为12个月,其中3个月为丹佛神经学试验评估,6个月为脑脊液和血清血清学调查。在同一天从患有艾滋病脑病的儿童身上收集成对的CSF和血清样本,分析血脑屏障(BBB)的渗透性和鞘内抗HIV特异性抗体的产生。我们进行了前瞻性随访,反复比较了WB谱和脑脊液和血清中抗V3抗体的存在,并评估了脑脊液抗体特异性(抗gag Western Blot评分)随疾病进展的变化。尽管血脑屏障通透性没有改变,但所有受试者鞘内IgG合成均有所增加。血清抗呕吐评分稳定在9.1 ~ 10.4之间,差异无统计学意义。相比之下,脑脊液样本的得分随着疾病的进展而显著下降,从无脑病儿童的8.7分,到静止性脑病组的6.5分和进展性脑病组的3.6分。ELISA测定的抗p24抗体水平与WB测定的抗gag评分之间存在很强的相关性。即使在免疫复合物解离后,p24抗原仅在3例中呈阳性。在功能性血脑屏障患者的脑脊液样本中未检测到抗V3抗体。脑脊液中抗gag抗体反应性的下降是疾病进展的早期指标,反映了严重的神经损伤过程。脑脊液样本中缺乏抗V3抗体,表明嗜神经毒株的PND在V3环的不同位置上映射。这些结果反映了在中枢神经系统中进化的抗原逃逸突变体的选择,不同于血淋巴性分离株。
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引用次数: 0
Observations regarding the neurocytokaryological study of the experimental infection with herpes simplex virus type 1 and type 2 in the white mouse. 1型和2型单纯疱疹病毒实验性感染小白鼠的神经细胞核学观察。
Pub Date : 1998-01-01
M Petica, I Crişan, A Muţiu

Mice distributed in groups were inoculated with the herpes virus simplex type 1 and type 2 strains in various dilutions (10(-1)-10(-7)), which had been maintained on primary human embryo cell cultures. The animals were killed and homogenates were prepared of their brains, which were used for testing the infective titre in vivo and for the isolation of nuclei, by means of the Hymer-Kuff technique. For the cytokaryological examinations, smears were prepared from the homogenate, which were stained by May-Grünwald-Giemsa and Mann methods. The cell and nucleus types were evaluated morphologically, descriptively, according to the size and structure. On the basis of three criteria the following groups were set up: large sized cells with bulky nuclei; middle sized cells with nuclei of a medium size; small sized cells with small nuclei and visible nucleoli; small sized cells with small, tachychromatic nuclei; cells exhibiting karyolysis. In both the infection with herpes virus simplex type 1 and type 2, the small sized cells with small tachychromatic nuclei represent the highest rate. Between the variants of the elaborated models no significant differences regarding the cell and nucleus types are recorded. On the whole, the cytokaryological picture does not exhibit notable differences, but rather resemblances with respect to the proportionality of the nucleus types in the experimental variants studied, in the case of herpes simplex virus type 1 and 2 induced infections.

小鼠分组接种不同稀释度(10(-1)-10(-7))的单纯疱疹病毒1型和2型菌株,这些菌株在原代人胚胎细胞培养中保持不变。杀死动物,制备脑匀浆,用Hymer-Kuff技术测定体内感染滴度和分离细胞核。细胞核学检查,从匀浆中制备涂片,用may - gr nwald- giemsa法和Mann法染色。细胞和细胞核类型根据大小和结构进行形态学、描述性评价。按3项标准分为大细胞、大细胞核组;中等大小的细胞,细胞核中等大小;细胞体积小,细胞核小,可见核仁;小细胞,细胞核小,快染;表现出核溶解的细胞。在1型和2型单纯疱疹病毒感染中,具有小快染性细胞核的小细胞比例最高。在详细阐述的模型的变体之间,没有记录到关于细胞和细胞核类型的显着差异。总的来说,在1型单纯疱疹病毒和2型单纯疱疹病毒诱导感染的情况下,细胞核图并没有表现出显著的差异,而是在研究的实验变体中,在核类型的比例上表现出相似之处。
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引用次数: 0
Virological and pathomorphological aspects in experimental infections with some triassociated pneumotropic viruses in the white mouse. 实验性小鼠嗜肺病毒感染的病毒学和病理形态学研究。
Pub Date : 1997-01-01
M Petica, A Petrescu

Experimental infections were induced in white mice by intranasal administration of parainfluenza virus type 3, 739-2D strain, to which influenza virus A/Beijing, 353/89 (H3N2) strain, and respiratory syncytial virus, Long strain, were associated. The model was organized so as to obtain a triassociated infection, parainfluenza virus type 3 being inoculated the first and the other two viruses, in the following stages-II and III-, alternately. The infections were revealed by the presence of positive immunofluorescence reactions in the pulmonary tissue, of histological, histochemical and histoenzymatic lesions at the level of the respiratory apparatus, as well as of pathomorphological changes in other organs. At the pulmonary parenchyma level the inflammatory lesion had a 100% frequency. The severest pathomorphological picture was the diffuse, lymphohistiocytic and macrophagic bronchopneumonia. The cytoinfiltrate was characterized by a proportionality between lymphocytes and histiocytes and the lower but not insignificant presence of macrophages. The prevalent lesion was the thickening of interparietoalveolar septa, as a consequence of stasis hyperemia, oedema and lymphohistiomacrophagocytic infiltrate, to which dystrophic lesions, especially of biocytes, are often added. Different lesions are present in other organs, such as: in the liver-inflammatory, dystrophico-inflammatory and dystrophic lesions; in the kidney, pancreas, cerebellum and thymus--vascular changes, stasis hyperemia; in the spleen--hyperplasia of megakaryocytes. Generally, the type and severity of lesions vary from one viral-association-induced infection to another.

用鼻内注射副流感病毒3,739 - 2d株诱导小鼠实验感染,其中与流感病毒A/Beijing, 353/89 (H3N2)株和呼吸道合胞病毒Long株相关。该模型被组织为三相关感染,在接下来的阶段- ii和III-中交替接种第一种副流感病毒和其他两种病毒。感染表现为肺组织免疫荧光阳性反应,呼吸器官的组织学、组织化学和组织酶病变,以及其他器官的病理形态学改变。在肺实质水平,炎性病变的发生率为100%。最严重的病理形态表现为弥漫性、淋巴组织细胞性和巨噬性支气管肺炎。细胞浸润的特征是淋巴细胞和组织细胞成比例,巨噬细胞的存在较少但并非微不足道。最常见的病变是肺泡壁间隔增厚,这是瘀血、水肿和淋巴组织巨噬细胞浸润的结果,通常还会出现营养不良病变,尤其是生物细胞。不同的病变存在于其他器官,如:在肝脏炎性、营养不良炎性和营养不良病变;肾、胰、小脑、胸腺——血管改变,瘀血充血;脾脏巨核细胞增生。通常,病变的类型和严重程度因病毒相关感染而异。
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引用次数: 0
Spectral and biological characteristics of some Acyclovir salts. 一些无环鸟苷盐的光谱和生物学特性。
Pub Date : 1997-01-01
C N Zaharia, I Crişan, D Rojanschi, A Muţiu

In the present work there are described some spectral characteristics in the ultraviolet and infrared region under various environment conditions for a series of synthetic analogues of deoxyguanosine, substances possessing antiviral properties. The study is performed on the Acyclovir compounds and on its Na, K and Li salts, synthetized in Romania, in comparison with the similar product Zovirax, of the "Wellcome" firm (England), used in the clinical practice for several years. The results show a very marked resemblance of the spectral behavior for all these products, a conclusion confirmed by the similar biological effects in the herpetic infection.

本文描述了一系列具有抗病毒作用的脱氧鸟苷类似物在不同环境条件下的紫外和红外光谱特征。该研究对在罗马尼亚合成的阿昔洛韦化合物及其钠、钾和锂盐进行了研究,并与在临床实践中使用了数年的“威康”公司(英国)的类似产品Zovirax进行了比较。结果表明,所有这些产物的光谱行为具有非常显著的相似性,这一结论在疱疹感染中具有相似的生物学效应。
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引用次数: 0
Cellular effects induced by some synthetic prostaglandins obtained at ICCF-Bucharest. 在ICCF-Bucharest获得的一些合成前列腺素诱导的细胞效应。
Pub Date : 1997-01-01
A Pleşa, F Cocu, R Repanovici

Four prostaglandin compounds synthetized at ICCF [chloprostenol triacetate (a), the isopropylic ester of the chloprostenol analogue (b), a PGA2 sulprostone analogue (c) and a PGE2 analogue (d)] were tested with respect to the cytotoxic effect, by successive studies on chicken embryo fibroblasts (CEF) and on human cells (HeLa). It was established that (b) and (d) display a high cytotoxicity, while (a) and (c) were relatively well tolerated by the cell layer. The spectrofluorometric determinations showed that these drugs did not induce modifications at the cell membrane level. The treatment with 4 micrograms/ml PGs (a) and (c) of HeLa cells inoculated with Sendai virus produced a significant decrease of TCID50/ml 24 hrs after drug introduction into the culture medium, from 10(6.5) to 10(4.5) and to 10(3.25), respectively.

通过对鸡胚成纤维细胞(CEF)和人细胞(HeLa)的连续研究,研究了在ICCF合成的四种前列腺素化合物[氯前列腺醇三醋酸酯(a),氯前列腺醇类似物的异丙酯(b), PGA2磺胺酮类似物(c)和PGE2类似物(d)]对细胞毒性的影响。(b)和(d)表现出较高的细胞毒性,而(a)和(c)对细胞层的耐受性相对较好。荧光光谱测定表明,这些药物在细胞膜水平上没有引起修饰。4微克/ml pg (a)和4微克/ml pg (c)处理仙台病毒接种的HeLa细胞,在药物进入培养基24小时后,TCID50/ml显著降低,分别从10(6.5)降至10(4.5)和10(3.25)。
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引用次数: 0
Experimental study of the possible implications of some apoptosis mechanisms in the pathogenesis of HIV infection. 1. In vitro apoptotic death of peripheral blood lymphocytes. 一些细胞凋亡机制在HIV感染发病机制中的可能意义的实验研究。1. 外周血淋巴细胞的体外凋亡。
Pub Date : 1997-01-01
F Topârceanu, C T Iucu, F Bârnaure

The human immunodeficiency virus (HIV) infection produces a gradual depletion of T-helper CD4+ lymphocytes as, surprisingly, a consequence of apoptosis of the uninfected lymphocytes. We suggested that this is the result of the action exerted by HIV inductors of apoptosis (for example, gp 120) in the absence of viral apoptosis suppressor, which confers protection to the infected cell. We intended to demonstrate this hypothesis within the framework of a complex study regarding the apoptosis mechanisms in HIV infection. We started this study by setting up an apoptosis model on HIV-negative peripheral blood lymphocytes (PBLs)* cultivated in vitro in the presence of dexamethasone (Dex). In this work we characterize this model morphologically and biochemically. Three unreported morphological changes observed by us--namely: I) fringing of nucleus with advancement of fringes up to the plasma membrane; II) segmentation and peripheral migration of condensed chromatin through a rotation movement; III) "flowering of the cell" consisting in the radial separation of the lymphocyte into centrally united "petals" with the tendency to form apically multiple apoptotic bodies--completed the classical morphology of the apoptosis phenomenon. The apoptotic death was confirmed by the oligonucleosomal (multiples of 200 bp) and mononucleosomal fragmentation of DNA isolated from lymphocytes.

令人惊讶的是,人类免疫缺陷病毒(HIV)感染导致t辅助CD4+淋巴细胞逐渐耗竭,这是未感染淋巴细胞凋亡的结果。我们认为,这是在缺乏病毒凋亡抑制因子的情况下,HIV细胞凋亡诱导剂(例如gp 120)发挥作用的结果,而病毒凋亡抑制因子可以保护受感染的细胞。我们打算在一项关于HIV感染中细胞凋亡机制的复杂研究框架内证明这一假设。我们首先建立体外培养的hiv阴性外周血淋巴细胞(pbl)*在地塞米松(Dex)作用下的凋亡模型。在这项工作中,我们在形态学和生物化学上表征了这种模型。我们观察到三种未报道的形态学变化,即:1)细胞核呈边缘状突起,边缘向上延伸至质膜;II)凝聚染色质通过旋转运动进行分割和外周迁移;3)“细胞开花”是指淋巴细胞径向分离成中央统一的“花瓣”,并倾向于在顶端形成多个凋亡小体——完成了细胞凋亡现象的经典形态学。从淋巴细胞中分离的寡核体(200 bp的倍数)和单核体DNA片段证实了细胞凋亡。
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引用次数: 0
Clinical manifestations in the West Nile virus outbreak. 临床表现为西尼罗病毒暴发。
Pub Date : 1997-01-01
E Ceauşu, S Erşcoiu, P Calistru, D Ispas, O Dorobăţ, M Homoş, C Bărbulescu, I Cojocaru, C V Simion, C Cristea, C Oprea, C Dumitrescu, D Duiculescu, I Marcu, C Mociorniţă, T Stoicev, I Zolotuşca, C Calomfirescu, R Rusu, R Hodrea, S Geamai, L Păun

During the summer of 1996 an unusual clustering of meningoencephalitis cases was recorded in the Capital City, Bucharest, and in some areas from South-East Romania. After an initial suspicion of an enteroviral etiology was discarded, the West Nile etiology was confirmed by specific antibodies demonstration through hemagglutination-inhibition and ELISA tests. This study included 251 patients with the diagnoses of West Nile acute encephalitis (166 cases), acute meningitis (57 cases) and acute febrile disease (33 cases). The patients' age ranged from 1 to 89 years (mean 51.1 years). The most frequent clinical manifestations were: fever (95.7% of cases), cephalalgia (92.6%), stiffness of the neck (89.1%), vomiting (62.5%), marked asthenia (46.5%), myalgia (28.9%). In addition, patients with encephalitis exhibited: alteration of consciousness (89.2% of cases), tremor of extremities (40.4%), ataxia (44%), paralysis (15.1%). The fatality rate was 15.1% in acute encephalitis, 1.8% in acute meningitis and 0% in the acute febrile disease.

1996年夏季,在首都布加勒斯特和罗马尼亚东南部的一些地区记录了不寻常的脑膜脑炎聚集病例。在最初怀疑是肠道病毒病因后,西尼罗河病因通过血凝抑制和ELISA试验证实了特异性抗体。本研究纳入251例诊断为西尼罗急性脑炎(166例)、急性脑膜炎(57例)和急性发热病(33例)的患者。患者年龄1 ~ 89岁,平均51.1岁。最常见的临床表现为发热(95.7%)、头痛(92.6%)、颈部僵硬(89.1%)、呕吐(62.5%)、明显虚弱(46.5%)、肌痛(28.9%)。此外,脑炎患者表现为意识改变(89.2%)、四肢震颤(40.4%)、共济失调(44%)、瘫痪(15.1%)。急性脑炎病死率为15.1%,急性脑膜炎病死率为1.8%,急性温病病死率为0%。
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引用次数: 0
A high number of severe neurologic clinical forms during an epidemic of West Nile virus infection. 西尼罗病毒感染流行期间出现大量严重神经系统临床表现。
Pub Date : 1997-01-01
C Cernescu, S M Ruţă, G Târdei, C Grancea, L Moldoveanu, E Spulbăr, T Tsai

The serologic confirmation of more than 800 cases hospitalized during the viral meningoencephalitis epidemic caused by the West Nile virus (WNV) that affected the South-East of Romania during the summer of 1996 consolidated the case definition in over 80% of the patients admitted to the hospital with neurological impairments. Other clinical forms of the WN infection were reported only scarcely during the epidemic and were seroconfirmed at a lower rate (60%). IgM capture ELISA (MAC-ELISA) is a test of choice for the rapid diagnosis. The major advantage of MAC-ELISA procedure is the high probability of accurate diagnosis of WN infection when the test is performed only with acute serum or cerebrospinal fluid (CSF) specimens obtained while the patient is still hospitalized. Rapid diagnosis by MAC-ELISA is important for the institution of public health control, but the results obtained have also some predictive values. We report the serological patterns of 65 pairs of CSF and serum samples collected in the early days of neuroinfection for diagnostic purposes. An unexpected onset of the intrathecal specific humoral immune response before serum immunoglobulins synthesis was recorded in 25% of cases. For 14 patients with intrathecal onset of IgM synthesis, their records evaluated retrospectively showed a severe evolution. The presence of only IgM antibodies in CSF is a characteristic which matched with other laboratory variables described which predict poor evolution in viral encephalitis: pleocytosis, elevated protein concentration in CSF (> 100 mg per deciliter), hyponatremia (< 130 mmol per liter).

1996年夏季,西尼罗病毒(WNV)引起的病毒性脑膜脑炎在罗马尼亚东南部流行期间,对800多例住院病例进行了血清学确认,巩固了80%以上因神经损伤而入院患者的病例定义。在流行期间,很少报告其他临床形式的西尼罗菌感染,血清确诊率较低(60%)。IgM捕获ELISA (MAC-ELISA)是快速诊断的首选检测方法。MAC-ELISA程序的主要优点是,当仅对患者住院期间获得的急性血清或脑脊液(CSF)标本进行检测时,其准确诊断WN感染的概率很高。MAC-ELISA快速诊断对公共卫生管理机构具有重要意义,但其结果也具有一定的预测价值。我们报告了65对脑脊液和血清样本的血清学模式收集在早期的神经感染诊断的目的。在血清免疫球蛋白合成之前,有25%的病例发生鞘内特异性体液免疫反应。对于14例鞘内发生IgM合成的患者,回顾性评估他们的记录显示了严重的演变。脑脊液中仅存在IgM抗体的特征与预测病毒性脑炎进化不良的其他实验室变量相匹配:多细胞增多、脑脊液中蛋白浓度升高(> 100 mg / dl)、低钠血症(< 130 mmol / l)。
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引用次数: 0
Role of human papillomaviruses in the malignant transformation of keratinocytes. 人乳头瘤病毒在角化细胞恶性转化中的作用。
Pub Date : 1997-01-01
R Achim

Human papillomaviruses (HPVs) are considered the main etiological agent of cervical cancer and of its precursory lesions (CIN). The HPV transformation potential consists in the presence of two oncogenes, E6 and E7, which inactivate the tumour-suppressor proteins p53 and pRB of the host cell, inducing uncontrolled transformation and proliferation mechanisms. These effects were demonstrated by in vitro experiments.

人乳头瘤病毒(hpv)被认为是宫颈癌及其前兆病变(CIN)的主要病因。HPV的转化潜力在于存在两个致癌基因E6和E7,它们使宿主细胞的肿瘤抑制蛋白p53和pRB失活,诱导不受控制的转化和增殖机制。体外实验证实了这些作用。
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引用次数: 0
Degree of polymerization dependent nucleic acid adsorption on cellulose. 聚合程度依赖于核酸在纤维素上的吸附。
Pub Date : 1996-01-01
O L Hörer

Experimental data on bichromatic multiple light scattering enhanced absorbance (MLSEA) photometry at 260 and 350 nm of nucleic acid chromophores on dried chromatography paper disks, previously impregnated with RNA solutions are analyzed. The extrapolated X(O) value of the adsorbed chromophore mole fraction X (9, 10) and mainly its slope dX(O)/dlogD in terms of the logarithmic plot of Debye length D (A), are strongly dependent upon the polynucleotide degree of polymerization P, providing a new ultramicroanalytical, rapid and simple parameter for relative P measurements. New experimental evidence is brought for the two-step adsorption model of polyelectrolytes at charged interfaces.

分析了在260 nm和350 nm处对核酸发色团进行双色多重光散射增强吸收(MLSEA)光度测定的实验数据。吸附的发色团摩尔分数X(9,10)的外推X(O)值及其斜率dX(O)/dlogD(德拜长度D (A)对数图)强烈依赖于聚合度P,为相对P的测量提供了一种新的超显微分析、快速和简单的参数。为聚电解质在带电界面的两步吸附模型提供了新的实验证据。
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引用次数: 0
期刊
Romanian journal of virology
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