Analysis of the biological and molecular variability of the Polish isolates of Tomato black ring virus (TBRV)

Tomato black ring virus (TBRV) is an important pathogen infecting many plant species worldwide. The biological and molecular variability of the Polish isolates of TBRV was analyzed. The analysis was performed based on the symptoms induced by various isolates on test plant species as well as on phylogenetic relationships between isolates. Isolates differed in their host range and symptomatology. In addition, genetic variation among isolates was characterized by restriction fragment length polymorphism analysis and confirmed by sequencing. The phylogenetic analysis revealed that the Polish isolates differ from each other and do not form a monophyletic cluster. Finally, we identified and analyzed sequences of defective RNA forms arising from the TBRV genome.     

Detection of Pepino mosaic virus isolates from tomato by one-step reverse transcription loop-mediated isothermal amplification

Loop-mediated isothermal amplification (LAMP) of DNA is a novel technique for the amplification of DNA under isothermal conditions. For the first time, we applied this method to develop a simple and sensitive tool for the detection of Pepino mosaic virus (PepMV). PepMV is an emerging pathogen that causes yield and quality losses in tomato crops. Specific RT-LAMP primers for PepMV detection were designed based on triple gene block sequences. The reaction was performed in a single tube at 65 °C for 30 min. The RT-LAMP assay is easy to perform and inexpensive, and it may be applied in the rapid and specific diagnosis of PepMV.     

Single mutation converts mild pathotype of the Pepino mosaic virus into necrotic one

Pepino mosaic virus (PepMV) is a member of the Flexiviridae family, genus Potexvirus. PepMV isolates from the same genotype differ in the symptoms they induce in Datura inoxia and Solanum lycopersicum. Necrotic isolates cause necrosis on these plants whereas mild isolates are very often asymptomatic or induce mild mosaic. Sequence analysis of the genomes of mild (PepMV-P22) and necrotic (PepMV-P19) strains revealed that they differ by several nonsynonymous mutations. Eleven mutations are located in the RdRp coding region and one mutation is placed in the TGB3 coding region. To investigate whether these mutations affect the symptom induction, both PepMV-P22 and PepMV-P19 were mutagenized. As a result we found that the genetic determinant responsible for necrosis induction was amino acid 67 of TGB3. This showed that one point mutation is sufficient to alter the virus from mild to aggressive in tomato and D. inoxia.     

Inhibition of lipopolysaccharide-inducible nitric oxide synthase and IL-1beta through suppression of NF-kappaB activation by 3-(1'-1'-dimethyl-allyl)-6-hydroxy-7-methoxy-coumarin isolated from Ruta graveolens L

The Ruta graveolens L. plant is used in traditional medicine to treat a large number of diseases. The methanol (50%) extract of the whole plant was observed to inhibit the expression of inducible nitric oxide synthase (iNOS) and the cycloxygenase-2 (COX-2) gene in lipopolysaccharide (LPS)-induced macrophage cells (J774A.1, [Raghav, S.K., Gupta, B., Agrawal, C., Goswami, K., Das, H.R., 2006b. Anti-inflammatory effect of Ruta graveolens L. in murine macrophage cells. J. Ethnopharmacol. 104, 234-239]). The effect of whole plant extract on the expression of other pro-inflammatory genes such as tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-12, interferon-gamma (IFN-gamma) and the activation of nuclear factor-kB (NF-kappaB) were investigated in LPS stimulated macrophage cells. An active compound was isolated from this methanol extract by further solvent fractionation and reverse phase high performance liquid chromatography (RP-HPLC). The purified compound was identified as 3-(1'-1'-dimethyl-allyl)-6-hydroxy-7-methoxy-coumarin having IUPAC nomenclature of 6-hydroxy-7-methoxy-3-(2-methyl but-3-en-2yl)-2H-chromen-2-one by ESI-MS, MALDI, FT-IR and NMR. Effect of this purified compound was assessed on iNOS, COX-2 and various pro-inflammatory cytokine genes and was observed to inhibit both the protein and mRNA expression of iNOS and IL-1beta in LPS challenged macrophages. Electrophoretic mobility shift assay (EMSA) and Western blot analyses indicated that the plant extract and the isolated active compound blocked the LPS-induced activation of NF-kappaB through the prevention of inhibitor-kB (IkB) degradation. The purified compound also showed the anti-oxidant activity. The active compound at a dose of 40 mg/kg body weight was observed to inhibit the iNOS and IL-1beta gene expression significantly in endotoxin-induced inflammatory model of BALB/c mice. The low level of nitric oxide production was also observed in the sera of compound treated mice. The normal behavioral condition in LPS challenged BALB/c mice was noticed when these were treated with active compound.     

Role of Glycoproteins Isolated from Epicoccum purpurascens in Host-Pathogen Interaction.

OBJECTIVE: Attachment to host matrix is an important provisory step for the institution of any fungal infection. The present study investigates the role of glycoproteins of Epicoccum purpurascens in host-fungal adherence. METHODS:Epicoccum spore-mycelial extract was fractionated on a concanavalin A-Sepharose column. Three glycoproteins of 12, 17 and 33 kDa (Epi p 1) were electroeluted and checked for hemagglutination and hemagglutination inhibition. The monosaccharide content of the highly potent protein Epi p 1 was determined by high-performance anion exchange chromatography and pulsed amperometric detection. The interaction of Epi p 1 with mannose-binding lectin (MBL) leading to the activation of the complement system was studied by immunoblot, ELISA and ELISA inhibition techniques. Immunoblot and immunoblot inhibition were carried out with culture filtrate to determine the nature of Epi p 1. RESULTS: 33 (Epi p 1)-, 17- and 12-kDa proteins were 58, 46 and 38 times more potent than crude extract in hemagglutination activity (HA). The HA of Epi p 1 was inhibited by N-acetyl glucosamine, glucose and laminin. Epi p 1 had a high mannose content, showed MBL binding in ion-dependent manner and caused complement activation. The protein was detected in culture filtrate and thus seems to play a significant role in fungal invasion. CONCLUSION: Epi p 1, an allergenic glycoprotein of E. purpurascens, is involved in host-fungal interactions through MBL.     

Role of scaphoid in the abduction and adduction movements of wrist joint

Being a carpal bone scaphoid has an important role in wrist movements. Wrist joint is a synovial modified ellipsoid joint where movements like flexion, extension and adduction, abduction take place around two axes (transverse and anteroposterior). These movements at the wrist joint are associated with considerable range of movements at the midcarpal joint, as same group of muscles act on both of these joints. A study has been done amongst 120 persons at the Institute of Postgraduate Medical Education and Research, Kolkata, West Bengal during the period from 1998-2000 to detect the important movements of scaphoid bone specially during the abduction and adduction of wrist joint (which occur in association with the intercarpal joints) and also to detect whether such movements have any speciality in the population of eastern part of India. It was found in this study that the scaphoid acts as a link bone between the two rows of carpal bones and prevents the buckling of midcarpal joint specially of the capitato-lunate joint interface.     

会阴穿支皮瓣的显微解剖研究和临床应用初步报告

目的本研究旨在通过对会阴区域的穿支血管定性和定量分析，即确定主要穿支血管的位置、数量、口径、蒂的长度类型、来源血管以及穿支所供应皮肤的面积，着重研究穿支皮瓣的血管解剖基础，为临床设计应用穿支皮瓣提供解剖学依据。方法选用经10％甲醛溶液固定、红色乳胶经桡动脉及髂外动脉联合灌注的成年尸体11具（22侧）。肉眼解剖，微细结构在10倍手术放大镜下观察，用游标卡尺（精确到0．2mm）测量。在尸体标本上，模拟临床皮瓣切取方式，对皮瓣内包含的血管、神经进行观测。结果我们发现该区直接与间接穿支血液供应丰富，其中以阴部外浅动脉腹股沟和会阴穿支、阴囊（唇）后动脉外侧穿支、闭孔动脉前皮支穿支四支最具临床意义，并且位置均比较恒定，分别位于皮瓣上、中、下方。上述四支穿支以及它们的伴行静脉彼此存在广泛的吻合，并在内收肌内侧以上的深筋膜层上方形成会阴区上方、中间、下方三组链式血管吻合网。另外还有分布于皮瓣上、中、下的四支重要的皮神经：生殖股神经股皮支、髂腹股沟神经皮支、阴囊（唇）后神经皮支、股后皮神经会阴支皮。结论会阴穿支皮瓣血供充足，静脉回流丰富、又具有神经支配及淋巴回流系统。切取方便，供区隐蔽，修复、重建会阴部外形及神经感觉均可以取得理想结果，是一种值得推广的方法。     

阴股沟皮瓣的显微解剖学基础研究

目的 明确阴股沟皮瓣的显微解剖基础.方法 对11具(22侧)成人尸体阴股沟区进行显微解剖学研究.结果 阴股沟区动脉血液供应充足,并且位置均比较恒定,其中以阴部外浅动脉和阴囊后动脉外侧支最为恒定.静脉回流丰富,神经支配可靠.结论 阴股沟皮瓣血运丰富,解剖简便易行,切取方便,供区隐蔽,再造性器官外形及神经感觉均较理想,是一种值得推广的方法.     

Anti-MBL autoantibodies in patients with rheumatoid arthritis: prevalence and clinical significance

Occurrence of autoantibodies in patients' sera is the characteristic feature of autoimmune disorders. We assessed the presence of anti-mannose binding lectin (MBL) autoantibodies in the sera of 107 rheumatoid arthritis (RA) patients and 121 control subjects by enzyme immunoassay. Elevated levels of anti-MBL autoantibodies in the sera of RA patients (P<0.0001) was detected for the first time. The ratios of anti-MBL positive in RA patients and controls were respectively 60.7% and 1.65%. Experiments were then designed to understand the functional relevance of these autoantibodies. An inverse correlation of anti-MBL autoantibodies with serum MBL levels (P=0.001) and MBL complex activity (P=0.02) was observed without genetic association between MBL polymorphisms and anti-MBL autoantibody secretion. A significant increase (P=0.038) in the level of anti-MBL autoantibodies was observed in 23 synovial fluid samples in comparison to the serum samples. Moreover, the anti-MBL autoantibodies were found to be more often present in the sera of RA patients (60.75% sensitivity, 98.35% specificity and 0.913 area under the ROC curve) in comparison to the IgM and IgG isotypes of rheumatoid factors (RF). Anti-MBL autoantibodies were still positive in 25.23% RA patients when both the RF isotypes were negative. Also, in RA patients, at all stages of disease activity and joint deformity, anti-MBL autoantibodies were more often present than both the RF isotypes. Therefore, the significant presence of anti-MBL autoantibodies enunciates that anti-MBL autoantibodies might have a diagnostic value; however, more studies are needed to confirm the role of anti-MBL autoantibodies in the diagnosis of rheumatoid arthritis.     

彩色多普勒能量图(CDE)检测乳腺癌血管生成活性与MVD相关性研究及其与转移的关系

目的：研究血管生成活性与肿瘤转移的关系，探讨彩色多普勒超声预测乳腺癌预后的应用价值。方法：43例乳腺癌病例，腋淋巴结转移即阳性组17例，腋淋巴结无转移即阴性组26例。分别于术前行彩色多普勒超声检查，检测肿瘤特性。于术后以CD34单克隆抗体作其病理组织切片的免疫组织化学染色，在显徽镜下进行徽血管计数即徽血管密度（MVD）。计量资料采用t检验，计数资料采用X^2检验及秩和检验。结果：（1）阳性组乳腺癌组织的MVD值（49．2士25．5）（个／100倍）高于阴性组乳腺癌组织的MVD值（32．1士14．4）（个／100倍），二者差异具有显著性意义（P〈0．05）；随着组织学级别的增加，乳腺癌组织的MVD值显著增大（P〈0．05）。（2）CDE显示肿块血流信号阳性组较阴性组丰富，且与其组织学分级及腋淋巴结转移率有关。随着组织学级别的增加，肿块内血流信号丰富程度有增加的趋势；随着肿块血流信号的增多，其腋淋巴结转移率有增大的趋势，至N型时显著性增大（P〈0．05）。结论：乳腺癌的血管生成活跃者，其腋淋巴结转移的可能性大。彩色多普勒能量图（CDE）技术的运用．有可能为临床预测乳腺癌预后提供简便、直观的形态学依据。