Subunit specific antisera to the Escherichia coli ATP synthase: effects on ATPase activity, energy transduction, and enzyme assembly

We obtained antisera to each of the five subunits (α, β, γ, δ, and ?) of the F₁ portion of the proton-translocating ATPase from Escherichia coli (ECF₁). No cross-reaction between the antiserum to a given subunit and any of the other four subunits was observed by Ouchterlony immunodiffusion. The α antiserum reacted only with the denatured α chain. Antibodies to either subunit β or subunit γ inhibited the ATPase activity of the enzyme. The ATPase activity of the holoenzyme in the everted membrane vesicles was just as sensitive as purified ECF₁ to inhibition by the anti-β or anti-γ serum. A prolonged digestion of ECF₁ with trypsin removed intact γ from ECF₁, but did not alter the sensitivity of the ATPase to inhibition by the anti-γ serum. Proteolytic fragments were isolated from the trypsinized enzyme. They gave an immunoprecipitation band with the anti-γ serum, but none of the other subunit antisera. The antiδ serum detached ECF₁ from everted membrane vesicles and completely blocked both the ATP- and respiration-dependent pyridine nucleotide transhydrogenase, an energylinked membrane function. The δ antiserum had no effect on the ATPase activity of the ECF₁. The e antiserum stimulated the ATPase activity of purified ECF₁ as shown previously (P. P. Laget and J. B. Smith, Arch. Biochem. Biophys.197, 83, 1979), but strongly inhibited the holoenzyme in membrane vesicles. The α antiserum completely blocked the ATP-driven transhydrogenase. The same antiserum maximally inhibited the respiratory chain-driven reaction by only 35%. These observations indicate that the antiserum selectively affected energy transduction mediated by the ATPase. The protonmotive force generated by substrate oxidation was probably not dissipated by the ? antiserum. Adsorbing the δ or ? antiserum with everted membrane vesicles selectively removed those antibodies that reacted with membrane-bound ATPase. The adsorbed sera still reacted strongly with purified ECF₁, and prevented it from restoring ATP-dependent proton translocation in ECF₁-depleted vesicles. Therefore, it appears that more of the δ and the ? subunit is exposed in the purified ECF₁ molecule than in the membrane-bound enzyme.     

Evolutionary Relationships Among <Emphasis Type="Italic">Aspergillus terreus</Emphasis> Isolates and their Relatives

Aspergillus terreus is a ubiquitous fungus in our environment. It is an opportunistic human pathogen and economically important as the main producer of lovastatin, a cholesterol lowering drug. Our aim was to examine the genetic variability of A. terreus and closely related species using molecular and analytical techniques. Lovastatin production was examined by HPLC. Lovastatin was produced by seven isolates belonging to the species A. terreus. RAPD analyses were carried out using 25 different random primers. Neighbor-joining analysis of RAPD data (120 characters) resulted in clustering of the A. terreus isolates into distinct groups. Some correlation was observed between lovastatin producing abilities of the isolates and their position on the dendrogram based on RAPD profiles. The internal transcribed spacer region and the 5.8S rRNA gene of A. terreus and related isolates was also sequenced. Phylogenetic analysis of sequence data let us classify the isolates into different clades which mostly correspond to the species Aspergillus terreus, Aspergillus flavipes, Aspergillus niveus, Aspergillus carneus and Aspergillus janus/A. janus var. brevis. Aspergillus allahabadii, A. terreus var. aureus and A. niveus var. indicus belonged to the A. niveus clade, while an Aspergillus isolate previously classified as A. niveus was most closely related to A. flavipes isolates. Aspergillus anthodesmis formed a distinct branch on the tree. Although it was previously suggested based on 28S rDNA sequence data that Aspergillus section Terrei should include A. carneus and A. niveus isolates, phylogenetic analysis of ITS sequences indicate that A. flavipes isolates are more closely related to A. terreus than A. carneus isolates. Our data suggest that sections Terrei and Flavipedes should be merged. However, further loci should be analysed to draw more definite conclusions.     

The effect of phencyclidine on dopamine metabolism in the mouse brain

The administration of phencyclidine (PCP) to mice resulted in no change in brain levels of tyrosine, dopamine (DA), norepinephrine (NE), or homovanillic acid (HVA). Although PCP reduced plasma tyrosine levels, no effect of PCP on the utilization of DA of NE after blockade of synthesis with α-methyl-p-tyrosine (AMPT) was observed. In addition, PCP did not affect the probenecid-induced accumulation of HVA. However, PCP was observed to potentiate the haloperidol-induced increase in HVA concentration, and the haloperidol-induced decline in DA levels after AMPT. The former effect was blocked by baclofen, suggesting that PCP mobilizes DA for impulse-dependent release. This effect could not be attributed to an antagonism of presynaptic DA receptors. These effects are similar to those of the “non-amphetamine” stimulant class of drugs.     

四川江油渔洞子剖面下三叠统飞仙关组底部龙介类的古生态意义

三叠系龙介类化石在欧洲早有报道。在我国四川江油渔洞子剖面下三叠统飞仙关组底部牙形刺Hindeodus pavus带微生物岩中首次发现环节动物龙介类化石Spirorbisphlyctaena,与以蓝菌为主的自养型底栖微生物群落共生,是早三叠世生物复苏的先驱者。     

贵州桐梓红花园剖面下奥陶统桐梓组灰岩微相和区域沉积分异

早奥陶世桐梓红花园剖面位于扬子区的内陆架沉积带,碳酸盐岩相的桐梓组浅滩序列中频繁出现强水动力环境下三叶虫、棘皮类、腕足类、砾屑和鲕粒单独或混合构建的沉积单元,有些滩相单元是在高能带风暴浪中快速堆积的,间或出现有云坪带沉积的白云岩。盐度异常和强水动力条件皆不利于后生动物造礁,仅见薄层钙质微生物席。通过与红花园之南的贵阳乌当以及之北的桐梓水坝塘剖面桐梓组的岩相学比较,厘定它们分别由蒸发云坪为主—高能滩为主—潮下带为主的相变序列构成,藉此划定桐梓组沉积期由南到北古地理背景差异性,认为朝北方向海水有逐渐变深的趋势,水深控制了白云岩化强度和海水能量指标。     

Heat and chill requirements of Fraxinus flowering in Galicia (NW Spain)

Fraxinus pollen data from eight Galician localities (1999-2003), recorded using 7-day Lanzoni VPPS pollen traps, were studied to determine their temporal and spatial distribution. The determination of the chill and heat required to trigger flowering and the start cumulative date were calculated using ten years of pollen data from Ourense. The sum of maximum temperatures from the 55 days before the peak date showed the lowest standard deviation coefficient and the mean quantity of accumulated heat was 741. Temperatures below 0°C and/or rainfall at the beginning of flowering caused a longer period before the peak date was registered. Temperatures recorded in November were very important for chill accumulation and determine the heat requirement needed to trigger Fraxinus flowering in Galicia.     

Preparation of highly infective Leishmania promastigotes by cultivation on SNB-9 biphasic medium

Protozoan hemoflagellates Leishmania are causative agents of leishmaniases and an important biological model for study of host-pathogen interaction. A wide range of methods of Leishmania cultivation on both biphasic and liquid media is available. Biphasic media are considered to be superior for initial isolation of the parasites and obtaining high promastigote infectivity; however, liquid media are more suitable for large-scale experiments. The aim of the present study was the adaptation and optimization of the cultivation of Leishmania promastigotes on a biphasic SNB-9 (saline-neopeptone-blood 9) medium that was originally developed for Trypanosoma cultivation and combines the advantages of biphasic and liquid media. SNB-9 medium is characterized with a large volume of the liquid phase, which facilitates the manipulation with the culture and provides parasite yields comparable to parasite yields on such liquid medium as Schneider's Insect Medium. We demonstrate that SNB-9 very considerably surpasses Schneider's Insect Medium in in vitro infectivity of the parasites. Additionally, we show that the ratio of apoptotic parasites, which are important for the infectivity of the inoculum, in Leishmania culture in SNB-9 is higher than in Leishmania culture in Schneider's Insect Medium. Thus, we demonstrate that the cultivation of Leishmania on SNB-9 reliably yields highly infective promastigotes suitable for experimental infection.     

中国蓼属头状蓼组植物叶表皮微形态及其分类学意义

采用光学显微镜对中国蓼属头状蓼组17种7变种植物的叶片下表皮微形态进行了观察研究,结果表明,其叶片下表皮微形态特征分为4种类型:(1)气孔器类型为无规则型,表皮细胞不规则形,垂周壁波状或深波状;(2)非典型不等型,偶有无规则型,表皮细胞多边形或不规则形,垂周壁弓形、波状或深波状;(3)平列型,表皮细胞不规则形,垂周壁深波状;(4)平列型兼有非典型不等型,表皮细胞不规则形,垂周壁波状。根据其叶片下表皮气孔器类型,结合该组植物形态、习性等特征,将中国蓼属头状蓼组植物划分为4个系,即掌裂叶系、多年生系、蓼子草系以及一年生直立系。     

老年股骨粗隆部不稳定骨折的治疗策略

目的:探讨老年患者股骨粗隆不稳定骨折的治疗方案。方法:选取近几年收治的56例老年粗隆部粉碎性不稳定骨折患者,分别采用了内固定和人工关节置换手术治疗,其中内固定36例,人工关节置换20例。结果:本组平均随访24个月,内固定组出现2例内置物断裂,再次人工关节置换,2例感染,1例清创后好转,1例取出内置物后创口方愈合;人工关节组1例术后关节脱位。术后harris评分内固定组75.6,人工关节组85.6。结论:老年患者,不同程度骨质疏松,较多都伴有不同程度的基础疾病,粗隆部粉碎骨折内置物选择及治疗难度大,人工关节置换对于功能康复及降低并发症等都有明显的优势。     

Nuclear DNA contents, rDNAs, and karyotype evolution in subgenus Vicia: III. The heterogeneous section Hypechusa

Summary. Nuclear DNA contents, automated karyotype analyses, and sequences of internal transcribed spacers from ribosomal genes have been determined in the species belonging to section Hypechusa of the subgenus Vicia. Karyomorphological results and phylogenetic data generated from the comparison of rDNA (genes coding for rRNA) sequences showed that sect. Hypechusa is not monophyletic; however, some monophyletic units are apparent (one including Vicia galeata, V. hyrcanica, V. noeana, and V. tigridis, another including V. assyriaca, V. hybrida, V. melanops, V. mollis, and V. sericocarpa), which partly correspond to morphology-based infrasectional groups. The relationships among these species and the species in sections Faba, Narbonensis, Bithynicae, and Peregrinae have been also investigated. Correspondence and reprints: Dipartimento di Agrobiologia ed Agrochimica, Università della Tuscia, via San Camillo de Lellis, 01100 Viterbo, Italy.