Development of Immune Response to Experimental Bovine Trichophyton vervucosum Infection

Abstract— Cell mediated and humoral immune responses to experimental Trichophyton verrucosum infection were assessed by sequential cutaneous biopsies, antibody assessments and microscopic monitoring of fungal presence. Histopathologic examination showed the accrual of lymphocytes and other inflammatory cells in the dermis of infected sites. Immunoperoxidase staining of frozen sections with monoclonal antibody preparations revealed an influx of macrophages, BoCD4+ and B0CD8+ lymphocytes and γδ T cells from the 5th day to the 33rd day of infection. A moderate influx of B cells was observed. Protein G-colloidal gold staining revealed the presence of immunoglobulins in the dermis and superficial epidermal layers. Trichophyton specific serum antibodies appeared between days 33 and 55. Microscopic assessment of infected tissues revealed an increase in T, verrucosum elements (mycelium and ectothrix spores) from days 19 to 55. Fungal elements in infected areas did not decrease until after both humoral and cell mediated elements of the immune response were established. These responses imply a combination of cell mediated and humoral events were associated with T. verrucosum immunity and clearance in the calf. Résumé— Le réponse immunitaire humorale et cellulaire a une infection expérimentale àT. verrucosum a été appréciée par des biospsies cutanées successives, des dosages d'anticorps et la recherche microscopique de champignons. L'examen histopathologique a montré un afflux de lymphocytes et d'autres cellules inflammatoires dane le derme des sites infectés. Les marquages en immunopéroxydase par un anticorps monoclonal de coupes congelées a montré un influx de macrophages, lymphocytes BoCD4+ et BoCD8+ et des cellules T γδ, du 5e au 33e jour de l'infection. Un marquage par une protéine G—or colloidal a révélé d'immunogolglobulines dans le derme et les couches supéerficielles de l'épiderme. Les anticorps spécifiques de Trichophyton sont apparus entre 33 et 55 jours. L'examen microscopique des tissus infectés a révélé une augmentation du nombre d'éléments de T. verrucosum (mycelium et spores ectothrix) du 19e au 55e jours. Les éléments fongiques dans les zones infectées n'ont pas diminué avant que les réponses humorales et cellulaires ne solent établies. Ces résponses impliquent qu'une coopération des réponses humorales et cellulaires étaient associées dans l'immunité et la défense contre T. verrucosum. Zusammenfassung— Die zellvermittelten und humoralen Immunantworten auf die experimentelle Infektion mit T. verrucosum wurden durch eine Serie von Hautbiopsien, Antikörperuntersuchungen und mikroskopischer Untersuchung auf das Vorhandensein von Pilzen ausgewertet. Die histopathologische Untersuchung zeigte eine Ansammlung von Lymphozyten und anderen Entzündungszellen in der Dermis der infizierten Stellen. Die Immunperoxidasefärbung der Gefrierschnitte mit monoklonalen Antikörperzubereitungen zeigte einen Influx von Makrophagen, BoCd4-und BoCD8-Lymphozyten und gamma-delta-T-Zellen vom 5. bis zum 33. Tag der Infektion. Es wurde auch ein mäßiger Influx von B-Zellen beobachtet. Die Protein-G-kolloidale Goldfärbung zeigte die Anwesenheit von Immunglobulinen in der Dermis und den oberflächlichen epidermalen Schichten. Trichophyton-spezifische Serumantikörper traten zwischen Tag 33 und 55 auf. Die mikroskopische Untersuchung infizierter Gewebe zeigte eine Zunahme von T. verrucosum-Bestandteilen (Myzel und exktothrixe Sporen) vom Tag 19 bis 55. Pilzteile in infizierten Bereichen verminderten sich weder, nachdem humorale, noch nachdem zellvermittelte Elemente der Immunantwort auftraten. Diese Reaktionen deuten an, daß eine Kombination von zellvermittelten und humoralen Vorgängen im Zusammenhang mit T. verrucosum-Immumtät und Abheilung beim Kalb vorliegt. Resumen Por medio de biposias cutáneas secuenciales, medida de anticuerpos y exámen microscópico de presencia de hongos, se estudió la respuesta inmunitaria de tipo humoral y celular producida por la infección experimental con T. verrucosum. El exámen histopatológico reveló la presencia de agregación de linfocitos y otras células inflamatorias procedentes de la dermis de los puntos infectados. La tintura por medio de inmunoperoxidasa de las secciones congeladas, con preparaciones monoclonales de anticuerpos, demostró un aflujo de macrófagos BoCD4 + y BoCD8 + linfocitos y linfocitos, Tαδ, desde el quinto hasta el día 33 la infección. También se observó un aflugo moderado de linfocitos B. La tintura aúrica de proteina coloidal G reveló la presencia de inmunoglobulinas en al dermis y capas superficiales de la epidermis. Los anticuerpos específicos para la especie Trichophyton aparecieron entre los días 33 y 55. El exámen microscópico de los tejidos afectados demostró un incremento, de los elementos füngicos T. verrucosum (micelio y esporas exótricas) desde los días 19 al 55. Los elementos fúngicos en áreas infectadas no disminuyeron hasta después del establecimiento de ambos tipos de respuesta inmunitaria, humoral y celular. Estas respuestas implican que la combinación de ciertos fenómenos de inmunidad celular y humoral, están relacionados con la desaparición y la inmunidad de la infección producia por T. verrucosum en el ternero.     

Computationally Assisted Structural Elucidation of Cembranoids from the Soft Coral Sarcophyton tortuosum

A persistent study on soft coral Sarcophyton tortuosum resulted in the characterization of two new cembranolides, tortuolides A and B (1 and 2), and a new related diterpene, epi-sarcophytonolide Q. Their structures were determined not only by extensive spectroscopic analysis but also by DFT calculations of ECD and NMR data, the latter of which was combined with statistical analysis methods, e.g., DP4+ and J-DP4 approaches. Anti-inflammatory and cytotoxicity activities were evaluated in this study.     

河南省樱桃谷鸭乙型肝炎病毒全基因组克隆与序列分析

对河南省樱桃谷鸭主产区信阳、新郑、焦作三地628份樱桃谷鸭血清样本应用PCR技术进行鸭乙型肝炎病毒（DHBV）检测,并将三地DHBV阳性样本各挑选1份进行DHBV全基因的扩增、克隆、测序及序列分析。结果显示,信阳、新郑、焦作三地樱桃谷鸭DHBV自然携带率分别为10.5%、8.9%、17.6%;3株DHBV基因组全长分别为3 021、3 027、3 024bp,均含有编码P、S和C蛋白的3个开放阅读框,各开放阅读框氨基酸同源性比较显示差异显著的区域位于P蛋白。序列分析显示,3株DHBV河南株核苷酸同源性为89.8%~93.9%,与参考株为89.4%~99.5%。遗传进化分析及P蛋白关键位点分析结果表明,信阳分离株为西方基因型,其余2株为中国基因型。本试验掌握了河南省樱桃谷鸭主产区DHBV自然感染情况,并成功克隆了3株樱桃谷鸭DHBV全基因序列,为该病毒的进一步研究提供了有益信息。     

Avermectins,a novel class of compounds: implications for use in arthropod pest control

对阿维菌素B_2a进行结构优化,在阿维菌素B_2a的C4\     

Role of NF-κB in inducing HEL cell proliferation and apoptosis during human cytomegalovirus infection

AIM:To investigate whether human cytomegalovirus(HCMV) regulate human embryonic lung fibroblast(HEL) cell proliferation and apoptosis by activating NF-κB.METHODS:Immunohistochemistry and Western blot analysis were used to detect the NF-κB translocation and/or Bcl-2 and the levels of I-κBα during HCMV infection. Apoptotic cell were examined by flow cytometry, and the HEL cell proliferation was determined by MTT.RESULTS:The levels of NF-κB in the nucleus reached highly 48 h postinfection, and the levels of I-κBα were low 24 h postinfection. The activity of NF-κB was inhibited 120 h postinfection. The levels ofbcl-2was accorded with the activity of NF-κB. HCMV promoted HEL cells to proliferate before 72 h postinfection and induced apoptosis 120 h postinfection.CONCLUSION:NF-κB plays a role in HEL cell proliferation and apoptosis during HCMV infection, and it involves in the pathological mechanisms of diseases associated with HCMV infection.     

In UV‐B Irradiated Rice High Levels of Mn Limits Chloroplasts Injury Triggered by Oxidative Stress

The molecular mechanism to control the oxidative burst exerted by Mn accumulation in rice (Oryza sativa L.) plants, grown in hydroponics containing 0.5, 2 and 8 mg l^–1 Mn and irradiated with a total biological effective UV‐B irradiation of 20.825 kJ m^?2, was investigated in the chloroplasts at the 15th and 21st days after germination. In both experimental periods, Mn accumulation kinetics in the leaves and in the chloroplast lamellae displayed overall increases. Coupled to higher membrane selectivity, superoxide production and acyl lipids peroxidation in the thylakoids decreased, prompting upper rates of the Hill and Mehler reactions. Connected to UV‐B irradiation, higher accumulated Mn in thylakoids was found to be chelated in a 36.5 kDa protein, with Mn/protein ratio of about 1 and high content of Gln, Asp, Glu, Leu and Gly, being its EPR spectrum characteristic of high‐spin Mn(II), in a S = 5/2 ground state. As this protein exhibited enzymatic catalysis of superoxide dismutation, it was concluded that, under UV‐B irradiation, the high internal tolerance of Oryza sativa L. to Mn during the vegetative growth also triggers the synthesis of a manganese protein that mimics superoxide dismutase functioning, therefore furnishing an additional intimate protection against oxidative stress.     

不同温度下气调及红桔油对嗜卷书虱的熏蒸作用研究

应用四元二次旋转组合设计,研究了温度,ＣＯ２,Ｏ２及红桔油四因子五水平不同组合对嗜卷书虱的熏蒸作用。结果表明四因子对嗜卷书虱熏蒸作用的效果影响显著;单因子效应分析表明,四因子中ＣＯ２浓度对试虫死亡率的影响最大,红桔油浓度次之,温度第三,Ｏ２浓度作用最小;因子间互作分析表明,温度和其它三个因子以及Ｏ２和红桔油间存在着极的交互效应,而ＣＯ２和Ｏ２以及红桔油间交互效应不显著。     

白头翁提取方法比较

本试验采用热回流和超声两种不同提取方法,以水及不同浓度的乙醇溶液为溶剂,制备10个样品,采用高效薄层色谱法、高效液相色谱法,通过比较得膏率、色谱图、白头翁皂苷B4峰面积等,考察提取方法对白头翁活性成分的影响。结果表明,两种提取方法中,热回流提取得膏率、白头翁皂苷B4得率均较超声提取法高;乙醇浓度对皂苷B4的提取有一定影响,热回流提取70%～80%乙醇提取率最高,超声提取50%～70%乙醇提取率最高。研究结果为白头翁提取方法的选用提供了科学数据。     

悬浮培养在口蹄疫疫苗中的应用

通过生物反应器中进行BHK21细胞悬浮培养并逐级放大,分别接种口蹄疫OJMS/2000株与Asia 1/JSL株,纯化灭活后制备50批疫苗,结果均符合《口蹄疫O型、亚洲I型二价疫苗（OJMS株＋JSL株）制造及检验规程》（以下称规程）所规定的各项标准,病毒146S抗原含量比转瓶培养提高10倍以上、疫苗的不良反应得到进一步改善。