Untersuchung der eiweißspindeln des kakteen-X-Virus mit fluoreszierenden Antikörpern

A?koliv na zá kladě mnoha pokus? se p? edpokládalo, ?e tzv. bÍlkovinná v?etena v buňkách tzn. buně?né inkluse X-viru kaktus? (Ca XV), jsou slo?ena z ?etních prodlou?ených ?ásti Ca XV, p?esto to dosud nebylo proká zá no. Proto jsme se pokusili pomocÍ fluoreskujÍcÍch protilátek doká ?at, ?e bilkovinná v?etena jsou skute?ně agregáty virových ?ástic. V těto práci jsme pouzili tzv. nep?Ímé metody. Nejprve jsme p? sobili na buňky obsahujÍci tato v?etena homologiokým antisé rem proti Ca XV, zÍskanym imunizacÍ králÍk? a teprve potom jsme buňky vlo?ili do roztoku fluoreskujicÍch protilátek proti králicimu γglobulinu. BÍlkovinná v?etena svitila potom ve fluorescen?nÍm mikroskopu silně ?lutozeleně (bylopou?ito fluoresceinisothiocyaná tu). Tato fluorescence ná m uká zala, ?e nastala pozitivnÍ reakce a ?e bÍlkovianá v?etena jsou slo?ena z virových ?ástic. ?etné kontrolnÍ pokusy potvrdily ná? základnÍ pokus.     

Factors determining the biosynthesis of griseofulvin and similar substances

Была высказана гипотеза, что биосинтез гризеофульвине, цитромицетина, фульвиновой кислоты, фусаeубине, руброфусарина и родствеооых им вешеств протекаеР не путем конденсации линейнон цепочки, а из двух более коротких цепочек, которые возникают конденсацией 4 и 3 ацетатных единп. реакции, осушествляющиеся на уровне этих цапочек (илн прнмежувочных структур) метиляция, редукция, и, возможно, последующая дегидратация или только эноэизация— определяют направление дальнeйшeй кондeнсации в такой стeпeни, что конeчныe продуктыотдичаются своeй структурой.     

Root exudates of plants

Summary The composition of the root exudates of barley and wheat in the initial growth phases was investigated; amino acids, organic acids, sugars and certain aromatic compounds could be identified. A knowledge of the composition of root exudates is important from the standpoint of the interaction between the plants and the micro-organisms in the rhizosphere. Some aspects of the rhizosphere effect connected with the present work are discussed.     

Enzymes of the jejunal enterochromaffine cells in man

Summary A study of enzymatic equipment of enterochromaffine cells (e.c.) in jenual biopsies obtained with a Crosby capsule in normal humans and patients with nontropical sprue was undertaken. The following enzymes were demonstrated: alkaline phosphatase and adenosine triphosphatase (cell membrane), acid phosphatase (corpuscular), non-specific esterase (diffuse and corpuscular, predominantly eserine resistant, in corpuscular localization E 600 resistant), DPN- and TPN-diaphorases and dehydrogenases of lactic acid, malic acid, isocitric acid, glucoso-6-phosphoric acid, succinic acid, -hydroxybutyric acid and -glycerophosphoric acid. Enzyme activities were not equal in all cells suggesting some type of secretory cycle. In most patients with untreated nontropical sprue or with the disease in relapse e.c. were more numerous and hypertrophic with elevated activities of non-specific esterase and acid phosphatase. Implications of these results are briefly discussed.With 8 Figures in the Text, of which 2 in Colour     

Preparation of anti-inhibitor serum and its effect on the multiplication of influenza virus

Antiserum был подготовлен противингибитор вируса haemagglutination (IVH)одна из эмбриона chorioallantoic мембр анныепутем иммун изации морских св инокЭто анти-инги битора в сыворотке заблокирован ингибирование IVH из chorioallantoicмембран в haemagglutination испытанияно ника кого влияния на ре цепторы эритроци товСыворотке кро] ви не влияет рост chorioallantoic мембраны клет ок инет нейтрализ овать воздействи е на вирус гриппа Она снизилась спо собность chorioallantoic мембр ан для adsorb вируса гри ппа, а также степен ь размножения вир усав такого рода тканиАвторы хоте ли бы поблагодари ть г-н Г. Ruttkay-Ne -палубе и для проведения эле ктрофореза меру ния.     

Retrovirus-mediated transgenic keratin expression in cultured fibroblasts: specific domain functions in keratin stabilization and filament formation.

With retrovirus-mediated gene transfer, we used intact and deleted keratin proteins to investigate the molecular basis of intermediate filament function. Three levels of assembly show a different stringency for the involvement of individual keratin domains: protein accumulation requires the alpha helix domains; stable filament formation additionally requires both N- and C-terminal domains of either one of the two interacting keratins, suggesting that head to tail homotypic interaction is important for effective elongation; and higher order organization of the cytoplasmic network depends on correct type I-type II pairing of keratins. The presence of two distinct interaction sites along potentially different axes may explain the characteristic morphology of keratin intermediate filament networks.     

Protease-induced infectivity of hepatitis B virus for a human hepatoblastoma cell line.

The human hepatoblastoma cell line HepG2 produces and secretes hepatitis B virus (HBV) after transfection of cloned HBV DNA. Intact virions do not infect these cells, although they attach to the surface of the HepG2 cell through binding sites in the pre-S1 domain. Entry of enveloped virions into the cell often requires proteolytic cleavage of a viral surface protein that is involved in fusion between the cell membrane and the viral envelope. Recently, we observed pre-S-independent, nonspecific binding between hepatitis B surface (HBs) particles and HepG2 cells after treatment of HBs antigen particles with V8 protease, which cleaves next to a putative fusion sequence. Chymotrypsin removed this fusion sequence and did not induce binding. In this study, we postulate that lack of a suitable fusion-activating protease was the reason why the HepG2 cells were not susceptible to HBV. To test this hypothesis, virions were partially purified from the plasma of HBV carriers and treated with either staphylococcal V8 or porcine chymotrypsin protease. Protease-digested virus lost reactivity with pre-S2-specific antibody but remained morphologically intact as determined by electron microscopy. After separation from the proteases, virions were incubated with HepG2 cells at pH 5.5. Cultures inoculated with either intact or chymotrypsin-digested virus did not contain detectable levels of intracellular HBV DNA at any time following infection. However, in cultures inoculated with V8-digested virions, HBV-specific products, including covalently closed circular DNA, viral RNA, and viral pre-S2 antigen, could be detected in a time-dependent manner following infection. Immunofluorescence analysis revealed that 10 to 30% of the infected HepG2 cells produced HBV antigen. Persistent secretion of virus by the infected HepG2 cells lasted at least 14 days and was maintained during several reseeding steps. The results show that V8-digested HBV can productively infect tissue cultures of HepG2 cells. It is suggested that proteolysis-dependent exposure of a fusion domain within the envelope protein of HBV is necessary during natural infection.     

Characterization of a calcium/calmodulin-dependent protein kinase homolog from maize roots showing light-regulated gravitropism

Roots of many species respond to gravity (gravitropism) and grow downward only if illuminated. This light-regulated root gravitropism is phytochrome-dependent, mediated by calcium, and inhibited by KN-93, a specific inhibitor of calcium/calmodulin-dependent protein kinase II (CaMK II). A cDNA encoding MCK1, a maize homolog of mammalian CaMK, has been isolated from roots of maize (Zea mays L.). The MCK1 gene is expressed in root tips, the site of perception for both light and gravity. Using the [³⁵S]CaM gel-overlay assay we showed that calmodulin-binding activity of the MCK1 is abolished by 50 M KN-93, but binding is not affected by 5 M KN-93, paralleling physiological findings that light-regulated root gravitropism is inhibited by 50 M KN-93, but not by 5 M KN-93. KN-93 inhibits light-regulated gravitropism by interrupting transduction of the light signal, not light perception, suggesting that MCK1 may play a role in transducing light. This is the first report suggesting a physiological function for a CaMK homolog in light signal transduction.Abbreviations CaM calmodulin - CaMK (II) Ca²⁺/calmodulin-dependent protein kinase (II) - CBP CaM-binding protein - CDPK Ca²⁺-dependent protein kinase - MCK1 maize homolog of mamalian CaMK This work is supported by the National Aeronautics and Space Administration grant No: NAGW 238.     

钙离子和蛋白激酶C对大鼠缺血海马脑片谷氨酸堆积的影响

采用大鼠海马脑片体外缺血模型观察钙离子和蛋白激酶Ｃ（ＰＫＣ）对神经元胞外谷氨酸（ＧＬＵ）堆积的影响,结果显示：海马脑片在体外“缺血”１０ｍｉｎ,ＧＬＵ在胞外的浓度增加４倍（从３２±４升高到１１３±１０ｐｍｏｌ／（ｍｉｎ．ｍｇＰｒ）．ｎ＝６）．Ｎ型钙通道拮抗剂蝙蝠葛苏林碱（ＤＳＬ）或无钙培养液均能有效抑制这种浓度的升高（Ｐ＜０．０１）．提示缺血１０ｍｉｎ引发的ＧＬＵ浓度升高是受Ｃａ２＋内流调控的．当脑片在缺血状况下孵育３０ｍｉｎ,ＤＳＬ只部分抑制这种ＧＬＵ堆积,而无钙培养液则无影响,但这额外的ＧＬＵ堆积可被ＰＫＣ抑制剂Ｈ－７完全阻断,而被ＰＫＣ激动剂ＰＤＢ所加强;且不受钙调蛋白抑制剂Ｃａｌｍｄａｚｏｌｉｕｍ和８－溴－ｃＡＭＰ影响．提示缺血３０ｍｉｎ,胞外ＧＬＵ的堆积受钙内流和ＰＫＣ双重调控。