The 3'-terminal primary structure of five eukaryotic 18S rRNAs determined by the direct chemical method of sequencing. The highly conserved sequences include an invariant region complementary to eukaryotic 5S rRNA.

The 3'-terminal sequences of 18S rRNA from chicken reticulocyte, mouse sarcoma, rat liver, rabbit reticulocyte and barley embryo were determined by the direct chemical sequencing method. The regions sequenced show complete homology for the first 73 nucleotides. A sequence 5'-proximal to the m6(2)Am6(2)A residues that is complementary to eukaryotic 5S RNAs is totally conserved. This supports the hypothesis that base-paired interaction between 5S and 18S rRNA, which are present in the large and small ribosomal subunits respectively, may be involved in the reversible association of ribosomal subunits during protein synthesis.     

The scale characteristics of two Aphanius species from southern Iran (Teleostei: Aphaniidae)

Fish scales are morphologically diverse among species, within species, and even among individuals. The present study investigated macro- and micromorphology of the scales between two closely related Aphanius species in southern Iran including Aphanius hormuzensis, collected from Gotab/Mehran River and A. stoliczkanus, from Howba hot sulphuric spring. The results indicated clear interspecific differences in scale macromorphology and microstructures. The scales of A. hormuzensis are characterised by a rounded shape, 7–9 (mean 8.0±0.83) primary radii, and a relatively large focus with oblong shape that positioned centrally on the scale, while the scales of A. stoliczkanus are characterised by a polygonal shape with few spines in posterior region, 8–11 (mean 10.0±0.78) primary radii and a small rounded focus that positioned posterocentraly on the scale. These species are morphologically close but clearly distinguished by their scale morphology and habitat requirements (e.g. water depth and food availability). Therefore, it can be assumed that clear differences in their scale morphology have been resulted by the combination of both genetic and environmental factors.     

Nucleoli and ribosomal RNA cistron numbers in Hordeum species and interspecific hybrids exhibiting suppression of secondary constriction

The interspecific hybrids of Hordeum exhibit selective suppression of secondary constriction formation in the chromosome(s) contributed by one of the two parents. A comparison of the number of SAT (secondary constriction) chromosomes in the metaphase cells and the maximum number of nucleoli in interphase cells revealed that the chromosomes capable of organising nucleoli were not always reflected through secondary constriction formation. — The rDNA (DNA complementary to rRNA) amounts were estimated by DNA-rRNA filter hybridisation in diploid and polyploid species of Hordeum and their hybrids. While similar rDNA proportions were present in diploid and autotetraploid lines of H. bulbosum, there were up to threefold differences between H. vulgare and allohexaploids. Furthermore, differences were also apparent between species of same ploidy level. Ribosomal RNA (18S+5.8S+26S) cistron numbers in each of the five experimental hybrids exhibiting the selective suppression of secondary constriction formation revealed no selective loss of rDNA. — The presence of a higher number of nucleoli than the number of SAT chromosomes seen and the presence of expected number of rRNA cistrons suggest that the suppression of secondary constriction formation is not due to selective loss of rDNA.     

Human immunodeficiency virus type 1 Nef protein inhibits activation pathways in peripheral blood mononuclear cells and T-cell lines.

Human immunodeficiency virus type 1 (HIV-1) Nef protein causes the loss of cell surface CD4 and interleukin-2 (IL-2) receptor (Tac) from peripheral blood mononuclear cells (PBMC) and CD4+ T-cell lines. As both CD4 and the IL-2 receptor play crucial roles in antigen-driven helper T-cell signalling and T-cell proliferation, respectively, the role of Nef in the viral life cycle may be to perturb signalling pathways emanating from these receptors. However, the intracellular targets for Nef that result in receptor down-regulation are unknown. Using a recombinant glutathione S-transferase-full-length 27 kDa Nef (Nef27) fusion protein, produced in Escherichia coli by translation from the first start codon of HIV-1 nef clone pNL4-3, as an affinity reagent to probe cytoplasmic extracts of MT-2 cells and PBMC, we have shown interaction with at least seven host cell protein species ranging from 24 to 75 kDa. Immunoblotting identified four of these proteins as p56lck, CD4, p53, and p44mapk/erk1, all of which are intimately involved in intracellular signalling. To assess the relevance of these interactions and further define the biochemical activity of Nef in signal transduction pathways, highly purified Nef27 protein was introduced directly into PBMC by electroporation. Nef27-treated PBMC showed reduced proliferative responsiveness to exogenous recombinant IL-2. Normally, stimulation of T-cells by IL-2 or phorbol 12-myristate 13-acetate provokes both augmentation of p56lck activity and corresponding posttranslational modification of p56lck. These changes were also inhibited by treatment of PBMC with Nef, suggesting that Nef interferes with activation of p56lck and as a consequence of signalling via the IL-2 receptor. Further evidence for Nef interfering with cell proliferation was the decreased production of the proto-oncogene c-myb, which is required for cell cycle progression, in Nef-treated MT-2 cells. In contrast to the binding characteristics and biological effects of Nef27, the alternate 25-kDa isoform of Nef (Nef25) produced by translation from the second start codon of HIV nef pNL4-3 (57 nucleotide residues downstream) was shown to interact with only three cellular proteins of approximately 26, 28, and 56 kDa from PBMC and MT-2 cells, one of which was identified as p56lck. Also, proliferation and posttranslational modification of p56lck in response to IL-2 stimulation were not profoundly affected by treatment of PBMC with Nef25 compared with Nef27.(ABSTRACT TRUNCATED AT 400 WORDS)     

Auxin signaling is closely associated with Zn-efficiency in rice (Oryza sativa L.)

This study elucidates the involvement of auxin with Zn-efficiency (ZE) in Zn-efficient rice var. Pokkali. Pokkali showed no significant decrease in morpho-physiological features, electrolyte leakage and total soluble proteins due to Zn deficiency as compared with Zn-sufficient seedlings. However, auxin inhibitor under Zn deficiency severely affected these characteristics, suggesting that ZE is associated with auxin signaling in rice. Results further revealed significant decreases in the expression of Zn transporter genes (OsIRT1, OsZIP4 and OsZIP1), OsDMAS1 (deoxymugeneic acid synthase) and phytochelatin in roots due to auxin inhibitor. It implies that auxin signaling may trigger Zn uptake, transport and chelation in rice seedlings to withstand Zn-deficiency. Further, significant reduction of major S-metabolites (cysteine, methionine, glutathione) and antioxidant enzymes (superoxide dismutase and glutathione reductase) along with increased H₂O₂ content, due to auxin inhibitor under Zn deficiency compared with controls. Taken together, these findings reveal that mechanisms associated with ZE in Pokkali are dependent on auxin signaling.     

Uptake and processing of biofilm and free-cell vaccines of Aeromonas hydrophila in indian major carps and common carp following oral vaccination--antigen localization by a monoclonal antibody

Uptake and processing of biofilm (BF) and free-cell (FC) vaccines of Aeromonas hydrophila were studied in the Indian major carps catla Catla catla, and rohu Labeo rohita and in the common carp Cyprinus carpio following a single dose oral vaccination of 10(11) CFU g(-1) fish. Fish were sampled at 0.5, 1, 3, 6, 12, 24 h and at 2, 3, 5 and 10 d following vaccination and antigen localization was studied in the gut, kidney and spleen employing monoclonal antibody based immunofluorescence and immunoperoxidase. The general pattern of antigen localization was similar in catla, rohu and common carp. Initially, both the BF and FC antigens were localized in the gut lumen, followed by their uptake by intraepithelial vacuoles and macrophages. Antigen administered orally was also seen in the spleen and kidney. Both BF and FC antigens were detected in the gut lumen of carp within 30 min following oral delivery. However, BF antigen remained in the lumen of the hindgut for 48 h compared to 6 h in the case of FC antigen. In the early stages, BF antigen was localized in the gut epithelial vacuoles while FC antigen was associated with the small macrophages of the hindgut. Antigen localization in spleen and kidney was observed at 3 h and persisted even up to 10 d following oral delivery. In general, there was a distinct difference between BF and FC vaccines in the duration of retention and quantity of uptake in the gut, kidney and spleen.     

Growth kinetics, protease activity and histophagous capability of Uronema sp. infesting cultured silver pomfret Pampus argenteus in Kuwait

Laboratory-produced and -reared sub-adults of silver pomfret (zobaidy) Pampus argenteus affected by severe scuticociliatosis during April and May 2005 were investigated and the causative was a scuticociliate, presumptively identified as Uronema sp. The parasite was capable of producing highly necrotic skin lesions. Mortalities started with a rise in the seawater temperature from 20 to 21.5 degrees C, coinciding with an increase in the total bacterial load of the rearing tank water from 10(3) to 10(5) CFU ml(-1). The parasite was successfully cultured in vitro (85 passages), using 10% brain heart infusion broth inoculated simultaneously with an aquatic Vibrio sp. The parasite reached peak cell density (1.82 x 10(5) cells ml(-1)) on the third day of inoculation. The parasite cell count was found to be inversely related (r = -0.45) to the bacterial count in the incubation medium. The ciliate body dimensions measured, on average, 34.05 +/- 0.99 microm in length and 16.70 +/- 0.74 microm in width (n = 300). Silver staining revealed morphological characteristics of Uronema sp., including a truncated anterior end, a buccal apparatus, caudal cilium and 3 oral polykinetids, with the first one appearing as a single row. Average number of kineties was 12 +/- 3. There was an inverse relationship (R2 = 0.84) between the proteolytic activity and the number of in vitro passages of the scuticociliate. However, the parasite from a freshly infected fish (using inoculum from the tenth passage) showed higher proteolytic activity (31.2%) compared with that produced by the inoculum from in vitro cultures (9.75%). The ciliate also showed a distinct histophagous capability when tested in vitro using host muscle tissue.     

Propagation of Arthropod-Borne Rickettsia spp. in Two Mosquito Cell Lines

Rickettsiae are obligate intracellular alphaproteobacteria that include pathogenic species in the spotted fever, typhus, and transitional groups. The development of a standardized cell line in which diverse rickettsiae can be grown and compared would be highly advantageous to investigate the differences among and between pathogenic and nonpathogenic species of rickettsiae. Although several rickettsial species have been grown in tick cells, tick cells are more difficult to maintain and they grow more slowly than insect cells. Rickettsia-permissive arthropod cell lines that can be passaged rapidly are highly desirable for studies on arthropod-Rickettsia interactions. We used two cell lines (Aedes albopictus cell line Aa23 and Anopheles gambiae cell line Sua5B) that have not been used previously for the purpose of rickettsial propagation. We optimized the culture conditions to propagate one transitional-group rickettsial species (Rickettsia felis) and two spotted-fever-group rickettsial species (R. montanensis and R. peacockii) in each cell line. Both cell lines allowed the stable propagation of rickettsiae by weekly passaging regimens. Stable infections were confirmed by PCR, restriction digestion of rompA, sequencing, and the direct observation of bacteria by fluorescence in situ hybridization. These cell lines not only supported rickettsial growth but were also permissive toward the most fastidious species of the three, R. peacockii. The permissive nature of these cell lines suggests that they may potentially be used to isolate novel rickettsiae or other intracellular bacteria. Our results have important implications for the in vitro maintenance of uncultured rickettsiae, as well as providing insights into Rickettsia-arthropod interactions.     

Pathogenesis of mulberry leaf spot in response to meteorological aspects of Kashmir,India

In this study, the influence of climatic factors on the development of leaf spot diseases of mulberry (Morus alba L.) in Kashmir, India was studied. It was found that the highest disease incidence and severity of Goshoerami variety was found in tree type (13.86% and 1.98%) followed by dwarf (7.53% and 1.67%) and bush (3.38% and 0.99%), respectively. Due to infection of pathogen, the relative water content and chlorophyll content of leaves were consistently decreased. The order of reduction in relative water content of infected leaves was found as: bush?>?dwarf?>?tree, while as the reduction in chlorophyll content: bush?>?tree?>?dwarf. It was also found that temperature of 25–30?°C and relative humidity of above 80% favoured the development of the disease. Rapid development of disease was found during the month of June, July and August when average temperature and relative humidity was about 25–30°C and 80%, respectively.