The function of the small subunits of ribulose bisphosphate carboxylase-oxygenase

When ribulose bisphosphate carboxylase-oxygenase from Synechococcus (strain RRIMP N1) was precipitated under mildly acidic conditions, most of its small subunits remained in solution. The precipitated enzyme readily redissolved at neutral pH and remained as an octamer of large subunits with some small subunits still attached. Optimum pH for this separation was 5.3 and disulfide-reducing reagents were not necessary. The fraction of small subunits removed by a single precipitation increased with increasing NaCl concentration. Catalytic activity of small subunit-depleted enzyme was linearly proportional to the fraction of small subunits remaining, while the carboxylase:oxygenase activity ratio and the affinity for CO2 remained constant. When isolated small subunits were added back to depleted enzyme preparations at neutral pH, reassociation occurred with return of catalytic activity. Under the usual assay conditions at pH 7.7, the binding constant of the small subunits was estimated to be about 10(-9) M. The small subunits also bound avidly to surfaces. However, loss of small subunits from the enzyme during the course of purification was minimal. The results are consistent with a simple model in which only those large subunits which have a small subunit bound to them are catalytically competent. Thus, an essential, even if indirect, role for the small subunits in catalysis is indicated.     

寒温带牧林交错区生境复杂度对啮齿类物种多样性的影响

采用铗日法对嫩江流域牧林交错区5个生境梯度中啮齿动物的多样性水平进行了研究,以探讨栖息地复杂程度对地面啮齿类物种多样性的影响.结果表明,黑线姬鼠、黑线仓鼠和大林姬鼠是嫩江流域牧林交错区中的优势种和常见种,不同物种在不同生境类型中呈明显的不均匀分布,反映了它们对不同生境的选择倾向性.同时,地面草本植物的覆盖度和丰富度对地面活动的小型哺乳动物的物种组成和群落结构具有较大的影响,啮齿动物的捕获率和生物量与草本植物的多样性水平呈现出显著的相关性关系,其物种丰富度随草本植物多样性指数的增加呈递增的趋势.此外,各生境中啮齿动物的物种多样性指数与草本植物多样性的相关关系并不显著,可能是因为灌丛区和林-灌交错区内大量的低矮灌丛与萌生丛改善了生境的隐蔽条件与食物资源,为更多物种的共存创造了良好的微环境,使得两生境中啮齿动物的物种多样性水平尤为突出.研究表明,小型啮齿动物的物种组成与栖息地的复杂程度有关,但地面植被对小型兽类生物量和物种多样性的影响力度并不是等同的,对地面小型兽类物种多样性影响的研究,统计生境复杂度或异质性时应考虑不同植被型/生境类型距地面的高度,不同植被型对小型兽类物种多样性的贡献程度随其距地面高度的增加而降低.     

浙江南部近海小黄鱼资源分布及其与环境因子的关系

根据2015—2016年浙江南部近海4个航次的底拖网资源调查数据,利用广义可加模型分析了调查期内小黄鱼资源的分布特征及其与环境因子的关系.结果表明:浙江南部近海的小黄鱼资源主要集中在鱼山渔场,夏季为小黄鱼资源的高产期,站点平均资源密度达到500.74 kg·h^-1·km^-2.不同季节影响小黄鱼资源密度及其分布的环境因子各不相同.其中,环境因子对秋季小黄鱼资源密度的影响效果并不显著.春季,小黄鱼主要分布于水深较浅的高盐水域;夏季,水温和盐度均与小黄鱼资源密度呈负相关关系,小黄鱼主要分布于中温高盐的鱼山海域;冬季,水温与资源密度呈正相关,小黄鱼栖息于水温适宜的外侧站点水域.总体上,小黄鱼资源的分布特征符合其洄游习性,但个别环境因子与资源密度的关系难以解释,仍需进一步研究.研究结果有助于了解浙江南部近海小黄鱼群体的生活习性,以及对小黄鱼资源的养护和管理.     

Biogenesis of small nucleolar ribonucleoproteins

Eukaryotic cells contain a very complex population of small nucleolar RNAs. They function, as small nucleolar ribonucleoproteins, in pre-ribosomal RNA processing reactions, and also guide methylation and pseudouridylation of ribosomal RNA, spliceosomal small nuclear RNAs, and possibly other cellular RNAs. Synthesis of small nucleolar RNAs frequently follows unusual strategies. Some newly discovered brain-specific small nucleolar RNAs of unknown function are encoded in introns of tandemly repeated units, expression of which is paternally imprinted. Recent studies of the protein components and factors participating in small nucleolar ribonucleoprotein assembly have revealed interesting connections with other classes of cellular ribonucleoproteins such as spliceosomal small nuclear ribonucleoproteins and telomerase. Cajal bodies emerge as nuclear structures important for the biogenesis and function of small nucleolar ribonucleoproteins.     

Global analysis of small RNA and mRNA targets of Hfq

Hfq, a bacterial member of the Sm family of RNA-binding proteins, is required for the action of many small regulatory RNAs that act by basepairing with target mRNAs. Hfq binds this family of small RNAs efficiently. We have used co-immunoprecipitation with Hfq and direct detection of the bound RNAs on genomic microarrays to identify members of this small RNA family. This approach was extremely sensitive; even Hfq-binding small RNAs expressed at low levels were readily detected. At least 15 of 46 known small RNAs in E. coli interact with Hfq. In addition, high signals in other intergenic regions suggested up to 20 previously unidentified small RNAs bind Hfq; five were confirmed by Northern analysis. Strong signals within genes and operons also were detected, some of which correspond to known Hfq targets. Within the argX-hisR-leuT-proM operon, Hfq appears to compete with RNase E and modulate RNA processing and degradation. Thus Hfq immunoprecipitation followed by microarray analysis is a highly effective method for detecting a major class of small RNAs as well as identifying new Hfq functions.     

Fine structure of small lymphocytes in the thymus of the mouse: Qualitative and quantitative analysis by electron microscopy

Summary Thymic small lymphocytes of dd-mice were qualitatively and quantitatively studied by electron microscopy. Differences in fine structure were revealed between cortical and medullary small lymphocytes. Cortical small lymphocytes are rounded in cell outline with a round nucleus. The cytoplasm surrounding the nucleus as a narrow rim is scanty and appears relatively dense due to an abundance of free ribosomes. The cell organelles are not well developed. On the other hand, medullary small lymphocytes are more irregular in shape with uneven cell membranes. Their nuclei are also more irregular in outline with frequent infoldings of the nuclear membrane. The cytoplasm is more abundant and paler with less numerous ribosomes. The cell organelles are better developed.Quantitative analysis was made of both cortical and medullary small lymphocytes by means of the point counting method. The nuclei of both cortical and medullary small lymphocytes are almost the same in size (a diameter of 4.9 ). The cell sizes are different between cortical and medullary lymphocytes: cortical small lymphocytes with a diameter of 5.5 were smaller than medullary ones with a diameter of 6.4 .Cortical small lymphocytes are very sensitive to the destructive effects of hydrocortisone, whereas the medullary ones are resistant.Periarteriolar lymphoid sheath in the splenic white pulp, which is known to be a thymus-dependent area, contains small lymphocytes that were similar in cytological details to medullary small lymphocytes of the thymus.In the light of the recent knowledge about a recirculating long-lived small lymphocyte pool, it appears probable that medullary small lymphocytes represent a contribution to the pool and that small lymphocytes with a long life span can be cytologically identified by electron microscopy.     

真菌小RNA的发生及其作用机制

小RNA是真核生物体内一种含量丰富的内源性非编码RNA,通过与其靶m RNA完全或非完全互补结合调控真核生物的基因表达。本文全面综述了真菌中已发现的小RNA种类、小RNA发生相关蛋白因子以及小RNA的具体作用机制,为进一步研究小RNA对真菌生长发育的调控机制提供重要参考。     

Quantitative study of deoxycytidine incorporation in large and small lymphocytes of the mouse

Using radioautographic smear preparations of thymocytes and mesenteric lymph node (MLN) cells labelled with three different tritiated pyrimidine deoxyribonucleosides, the incorporation of DNA precursors was studied separately on large lymphocytes and small lymphocytes. Radioautographic reaction due to generally tritiated deoxycytidine ( [G-3H]CdR) labelling in vivo in large lymphocytes was more intense than that in small lymphocytes. When mice were sacrificed 6 hr after the administration of tritiated thymidine ( [3H]TdR), small lymphocytes were labelled more heavily than large lymphocytes. However, labelling intensity with [3H]TdR in large lymphocytes was greatly enhanced by the administration of 5-fluoro-deoxyuridine, whereas in small lymphocytes labelling intensity was only fairly enhanced by the same treatment. When cells were incubated in vitro with 5-tritium labelled deoxycytidine [( 5-3H]CdR) for 10 min, there was no significant difference in labelling intensities between large and small lymphocytes. In the case of [G-3H]CdR incorporation, the labelling intensity in large lymphocytes was found to be significantly stronger than that in small lymphocytes. Large as well as small lymphocytes incorporated [3H]TdR very well in vitro. However, addition of 5 X 0 X 10(-5) M of non-radioactive CdR to the medium greatly decreased the incorporation of [3H]TdR by large lymphocytes, whereas the effect of non-radioactive CdR in small lymphocytes was not so marked as that in large lymphocytes. Furthermore, the [3H]TdR-labelling percentages were decreased at the same rate by the addition of non-radioactive CdR in both large and small lymphocytes. These results indicate that large lymphocytes and a proportion of small lymphocytes have a strong tendency to convert CdR to thymidine mono-phosphate, which is utilized for DNA synthesis, whereas this ability is relatively weak in the rest of small lymphocytes. Thus, it is probably that this metabolic ability changes during the transition of the large lymphocyte to the small lymphocyte.     

Population dynamics of large and small mammals

We offer an evaluation of the Caughley and Krebs hypothesis that small mammals are more likely than large mammals to possess intrinsic population regulating mechanisms. Based on the assumption that intrinsic regulation will be manifest via direct density-dependent feedbacks, and extrinsic regulation via delayed density-dependent feedbacks, we fit autoregressive models to 30 time series of abundance for large and small mammals to characterize their dynamics. Delayed feedbacks characterizing extrinsic mechanisms, such as trophic-level interactions, were detected in most time series, including both small and large mammals. Spectral analyses indicated that the effect of such delayed feedbacks on the variability in population growth rates differed with body size, with large mammals exhibiting predominantly reddened and whitened spectra in contrast with predominantly blue spectra for small mammals. Large mammals showed less variance and more stable dynamics than small mammals, consistent with, among other factors, differences in their potential population growth rates. Patterns of population dynamics in small versus large mammals contradicted those predicted by the Caughley and Krebs hypothesis.     

Species interactions and habitat associations of birds inhabiting urban areas of Sydney,Australia

Abstract The absence of small birds from many suburban areas may be due to adverse garden characteristics, interspecific aggression or human behaviour such as supplementary food provisioning that encourages predators. We investigated the relationship between these factors and the presence of seven small bird species in Sydney through a community‐based survey. The survey was conducted by participants over a 7‐day period between 7 am and 10 am in November and early December 2000. Three dominant species, the noisy miner (Manorina melanocephala), pied currawong (Strepera graculina) and common myna (Acridotheres tristis) were each present in over 59% of gardens. Each small bird species was present in less than 40% of gardens. All small birds were negatively associated with noisy miners, but only the silvereye (Zosterops lateralis) was negatively associated with pied currawongs. None of the species of small birds was negatively associated with common mynas. Four species of small birds were associated with at least one habitat variable, notably the proportion of native vegetation. Although more birds were recorded in gardens in which meat was provided, there were significantly fewer small birds in these gardens. There were also more birds recorded in gardens where seed was provided, with red‐browed finches (Neochmia temporalis) positively associated with seed provisioning in most regions of Sydney. The presence of dogs and cats was not related to the total abundance of birds overall or small birds in gardens. While garden characteristics may influence the presence of small birds to some degree, the presence of noisy miners, a species that are thought to aggressively exclude other species from their territories, is likely to be an important influence on these species in suburban areas. Furthermore, supplementary feeding by people is likely to negatively influence some small birds. The presence of carnivorous pets does not seem to influence the presence of small birds at the scale of the individual garden.     

Formation of the small subunit in the absence of the large subunit of ribulose 1,5-bisphosphate carboxylase in 70 S ribosome-deficient rye leaves.

Immunological tests with monospecific antisera to ribulosebisphosphate carboxylase (EC 4.1.1.39) and to its large and small subunits indicated the presence of a protein with antigenic properties of the small subunit in the absence of the large subunit in the leaves of young rye plants (Secale cereale L.) with a high-temperature-induced (32 °C) deficiency of 70 S plastid ribosomes. The small subunit-like protein was isolated from crude extracts of plastid ribosome-deficient 32 °C-grown leaf tissue by the use of columns with immobilized antibody. The main polypeptide retained by the immobilized antibodies had the same mobility after electrophoresis on sodium dodecyl sulfate-polyacrylamide gels as the small subunit of ribulosebisphosphate carboxylase and was also immunologically identical to the small subunit. The small subunit-like protein was present in the supernatant as well as in the membrane fraction of isolated 70 S ribosome-deficient plastids. At very young stages of normal leaves grown at a permissive temperature (22 °C) an excess of small subunit was observed that was also not integrated into the complete ribulosebisphosphate carboxylase molecule. From the results, we conclude that the synthesis of the small subunit occurs on cytoplasmic ribosomes and is not strictly coordinated with the translation of the large subunit in the chloroplast. During early leaf development, the formation of the large subunit seems to be the ratelimiting step in the synthesis of ribulosebisphosphate carboxylase.     

Dissociation of retinoblastoma gene protein hyperphosphorylation and commitment to enter S phase.

Mitogenic activities of simian virus 40 large T and small t antigens were studied in serum-deprived human diploid fibroblasts. Wild-type large T and small t cooperated in stimulating DNA synthesis and in inducing hyperphosphorylation of the Rb gene product (pRb). In contrast, a T antigen mutant defective for pRb binding (Rb- T) possessed no detectable mitogenic activity alone and failed to complement small t in stimulating DNA synthesis. Surprisingly, Rb- T and small t cooperated as strongly as wild-type T and small t with respect to pRb hyperphosphorylation. As a consequence, in two closely related conditions (i.e., stimulation by small t plus wild-type T versus small t plus Rb- T), the fraction of pRb in hyperphosphorylated forms dissociated from the fraction of cells in the S phase. These results indicate that pRb hyperphosphorylation is not always tightly coupled with a commitment to initiate DNA replication.     

Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5' terminal nucleotide

Argonaute (AGO) proteins recruit small RNAs to form the core of RNAi effector complexes. Arabidopsis encodes ten AGO proteins and a large network of small RNAs. How these small RNAs are sorted into specific AGO complexes remains largely unknown. We have cataloged small RNAs resident in four AGO complexes. We found that AGO2 and AGO4 preferentially recruit small RNAs with a 5' terminal adenosine, whereas AGO1 harbors microRNAs (miRNAs) that favor a 5' terminal uridine. AGO5 predominantly binds small RNAs that initiate with cytosine. Changing the 5' terminal nucleotide of an miRNA predictably redirected it into a different AGO complex and alters its biological activity. These results reveal a role for small RNA sequences in assorting among AGO complexes. This suggests that specialization of AGO complexes might involve remodeling the 5' end-binding pocket to accept certain small RNA sequences, perhaps explaining the evolutionary drive for miRNAs to initiate with uridine.     

The effect of habitat fragmentation on the bee visitor assemblages of three Australian tropical rainforest tree species

Tropical forest loss and fragmentation can change bee community dynamics and potentially interrupt plant–pollinator relationships. While bee community responses to forest fragmentation have been investigated in a number of tropical regions, no studies have focused on this topic in Australia. In this study, we examine taxonomic and functional diversity of bees visiting flowers of three tree species across small and large rainforest fragments in Australian tropical landscapes. We found lower taxonomic diversity of bees visiting flowers of trees in small rainforest fragments compared with large forest fragments and show that bee species in small fragments were subsets of species in larger fragments. Bees visiting trees in small fragments also had higher mean body sizes than those in larger fragments, suggesting that small‐sized bees may be less likely to persist in small fragments. Lastly, we found reductions in the abundance of eusocial stingless bees visiting flowers in small fragments compared to large fragments. These results suggest that pollinator visits to native trees living in small tropical forest remnants may be reduced, which may in turn impact on a range of processes, potentially including forest regeneration and diversity maintenance in small forest remnants in Australian tropical countryside landscapes.     

小细胞肺癌发生阻塞性肺炎相关因素分析

目的通过对小细胞肺癌患者是否发生阻塞性肺炎的相关因素对比分析,了解伴有阻塞性肺炎小细胞肺癌患者临床特点。方法采用回顾性分析方法,收集2003年1月至2013年1月大连医科大学附属第二医院所收治的经组织学或细胞学确诊的小细胞肺癌患者共450例。经核查资料后,进入统计学的伴有阻塞性肺炎组100例,不伴有阻塞性肺炎组112例,均为初治。对以下因素进行分析：性别、年龄、居住环境、吸烟情况、分期、肿瘤位置、病理类型、初始疗效、NSE（神经元特异性烯醇化酶）。计数资料率和构成比的比较采用卡方检验。结果本组研究中伴有阻塞性肺炎组患者100例中,吸烟患者占76%,小细胞未分化癌患者占81%,中心型癌患者占52%,广泛期患者占57%,NSE阳性率为92%,均高于不伴有阻塞性肺炎组患者。不伴有阻塞性肺炎组的小细胞癌患者初始疗效有效率明显高于伴有阻塞性肺炎组患者。两组在性别、年龄、居住环境方面差异无统计学意义。结论伴有阻塞性肺炎组的小细胞肺癌患者NSE阳性率高于不伴有阻塞性肺炎组。吸烟是小细胞肺癌患者发生阻塞性肺炎的高危因素。阻塞性肺炎降低小细胞癌患者化疗初始疗效。     

Pancreatic small cells: analysis of quiescence, long-term maintenance and insulin expression in vitro

We have previously identified a novel population of small cells in human and canine pancreas characterized by immature morphology, quiescence, and a glucose-responsive insulin secretion. Based on their immature phenotype and predominant presence in small islets, we have hypothesized that small cells serve as islet progenitors. This hypothesis remains untested, however, due to persistent quiescence and scarcity of small cells in vitro. We have recently developed a culture medium that allowed for modest small cell proliferation. In this study we characterized the expression of genes potentially involved in small cell growth regulation by Q-RT-PCR. Our results suggest that quiescence of small cells correlates with up-regulation of Cdk inhibitors p27(Kip1), p16(INK4a) and p21(CIP1), PTEN, Hep27 and Foxo1a and with down-regulation of c-Myc and the receptors for EGF, FGF2 and HGF. The exit from quiescence correlates with activation of EGFR expression and down-regulation of p27(Kip1) and p16(INK4a). We also report here that small cells can be maintained in long-term non-adherent cultures preserving insulin and glucagon production for up to 208 days. Therefore, expansion of small cells in vitro may have a significant potential for the treatment of diabetes. This study is an important step in understanding the mechanisms involved in small cell growth regulation, which is required to fully evaluate their functional potential.     

Localization of L-glutamate decarboxylase immunoreactivity in the major pelvic ganglion and in the coeliac-superior mesenteric ganglion complex of the rat

The localization of L-glutamate decarboxylase (GAD), the GABA-synthesizing enzyme, was studied in the rat major pelvic ganglion and in the coeliac-superior mesenteric ganglion complex by indirect immunofluorescence technique with a specific antiserum raised in rabbits. GAD immunoreactivity was demonstrated in small cells of these ganglia. The GAD-immunoreactive small cells were 10-20 microns in diameter and formed clusters or occurred as solitary cells. The principal neurons were non-reactive but they were surrounded by immunoreactive processes. Studies on colocalization of GAD with tyrosine hydroxylase (TH), the rate-limiting enzyme of the catecholamine synthesis, in the major pelvic ganglion and in the coeliac-superior mesenteric ganglion complex indicated that all GAD-immunoreactive small cells were also labelled with TH. In the major pelvic ganglion all TH-immunoreactive SIF cells were also immunoreactive for GAD. However, in the coeliac-superior mesenteric ganglion complex there occurred TH-immunoreactive small cells which showed no immunoreactivity to GAD. It is suggested that the small GAD-immunoreactive cells represent small intensely fluorescent (SIF) cells.     

The development of arylsulphatase in the small intestine of the rat

1. Arylsulphatase activity was measured in stomach, proximal and distal third of small intestine, colon, liver and kidney of foetal and neonatal Sprague-Dawley rats and Swiss mice, with nitrocatechol sulphate as substrate. 2. The specific activity in the distal small intestine, but not in the stomach, proximal small intestine or colon, increased about fourfold between 5 and 16 days after birth in both conventional and germ-free rats. 3. No comparable increase occurred in the distal small intestine of the mouse. 4. The specific activity of acid phosphatase in the distal small intestine of the rat rose only slightly when the arylsulphatase activity increased. 5. The pH optimum and Michaelis constant of arylsulphatase activity of the distal small intestine were similar for 1-day-old, 9-day-old and adult rats. 6. When extracts of distal small intestine of 1-day-old and 9-day-old rats were incubated together, the arylsulphatase activities were additive.     

Myoelectric response of the small intestine to the orad presence of the tapeworm Hymenolepis diminuta.

During its 24-hr migratory cycle in the small intestine, Hymenolepis diminuta is located in the orad part of the small intestine during the early morning hours and then in the caudad part of the small intestine during the late afternoon and early evening. During the later period, tapeworm-induced alterations of interdigestive myoelectric activity, a correlate of smooth muscle contraction or intestinal motility, are most intense in the ileal region. The hypothesis tested was that the tapeworm-induced changes in intestinal motility are local responses of the intestine responding to the close proximity of the lumenally positioned tapeworm and to the nutritional state of the host. The small intestine was monitored before and for 20 days after infection using electrodes implanted on the serosa of the small intestine. Myoelectric recordings were analyzed for the frequency of the normal patterns of interdigestive myoelectric spiking patterns and the altered myoelectric spiking related to tapeworm infection. During the morning hours, when the tapeworms are situated in the orad small intestine, no changes were observed during the normal myoelectric pattern of the digestive phase in any region of the intestine. When examined after the conversion of the digestive to interdigestive phase of motility, only on day 10 postinfection was the interdigestive phase significantly altered. It was concluded that the presence of the tapeworm in the orad small intestine during the satiety stage of the rat causes no changes in the electric events of the small intestine, with the exception of day 10 postinfection. Because tapeworms in the orad small intestine do not induce the tapeworm-altered myoelectric activity observed in the afternoon and evening with caudally positioned tapeworms, tapeworm-altered motility is not simply a response of the small intestine to the local presence of the tapeworm.     

Mutational analysis of delta antigen: effect on assembly and replication of hepatitis delta virus.

Hepatitis delta virus requires a helper function from hepatitis B virus for packaging, release, and infection of hepatocytes. The assembly of large delta antigen (HDAg) is mediated by copackaging with the small surface antigen of hepatitis B virus (HBsAg), and the assembly of small HDAg requires interactions with large HDAg. To examine the molecular mechanisms by which small HBsAg, large HDAg, and small HDAg interact, we have established a virion assembly system in COS7 cells by cotransfecting plasmids encoding the small HBsAg, the small HDAg, and large HDAg mutants. Results indicate that sequences within the C-terminal 19-amino-acid domain flanking the Cxxx isoprenylation motif are important for the assembly of large HDAg. In addition, a large HDAg mutant bearing extra sequences separating the C-terminal 19-amino-acid domain from the common regions of the small and large HDAgs is capable, like the wild-type large HDAg, of copackaging with small HBsAg. The ability of assembly is also demonstrated for a large HDAg mutant from which nuclear localization signals have been removed. Furthermore, a cryptic signal within the N-terminal 50 amino acid residues other than the putative N-terminal coiled-coil structure and a subdomain between amino acid residues 50 and 65 of the large HDAg are important for the assembly of small HDAg as well as the trans-dominant negative regulation of large HDAg in hepatitis delta virus replication.