Identification of Leptospira in Patients With Renal Failure Using Serological, Molecular and Pathological Based Techniques, in Shiraz, Iran

Leptospirosis is a relatively rare bacterial infection that affects people and animals caused by pathogenic species of Leptospira. The present study was conducted using nested polymerase chain reaction (PCR), microscopic agglutination test (MAT) and hematological and biochemical tests on 200 blood samples of renal disorders patients in Shiraz, Iran. Also nested-PCR assay and Warthin-Starry (WS) silver staining method was performed on 30 nephrectomised kidney sample. The frequency of pathogenic species of Leptospira infection in patients with renal disorders was 20 % and this infection was significantly correlated with BUN, anemia, RDW, MCV, MCH and hemoglobin levels (P < 0.01). MAT analysis showed that serum samples had positive titers against L. Grippotyphosa (13 samples), L. Ballum (6 sample), L. Pomona (3 samples), L. Canicola (2 samples), L. Icterohaemorrhagiae (1 sample) and L. Hardjo (1 sample) serovars. Twenty-three percent of the kidney samples from the patients with pyelonephritis were infected with the pathogenic species of Leptospira. This study showed that pathogenic Leptospira serovars are present in this area and in patients with renal disorders more attention should be paid to this zoonotic disease.

POLYMORPHISM OF TAGLGAP MICROSATELITE LOCUS AND ITS CONNECTION WITH ALLELELIC VARIETIES OF GLIADINS OF BREAD WHEAT

Introduction. Gliadins are monomeric and highly polymorphic storage proteins of wheat endosperm, which together with glutenins form a gluten complex that determines the breadmaking properties of wheat. Allelic variants of gliadins are an important feature in the selection of material for breeding, but their determination by electrophoresis in acid PAGE is quite difficult. Aim. The aim of this study was to investigate the polymorphism of the Taglgap microsatellite locus and to analyze its correspondence to the polymorphism of allelic variants of gliadins that have been revealed by acid PAGE electrophoresis. Methods. 140 cultivars and lines of bread wheat of Ukrainian and foreign selection were analyzed. Electrophoresis of storage proteins was performed in an acid PAGE according to the method of F. O. Poperellia (1989), allelic variants were designated according to the international nomenclature (Metakovsky et al., 2018). DNA was isolated by CTAB method and PCR was performed with primers to the Taglgap microsatellite (Devos et al., 1995). PCR products were fractionated in 7% PAGE and stained with silver staining method. Nucleotide sequences were searched by BLAST and aligned by MAFT methods. The main results. 19 allelic variants of gliadins and 11 alleles of the Taglgap locus were identified. In the collection of Ukrainian varieties there were Gli-B1b, Gli-B1c, Gli-B1d, Gli-B1e, Gli-B1f, Gli-B1g, Gli-B1h, Gli-B1l and Gli-B1o allelic variants and alleles of Taglgap 216 bp, 237 bp, 246 bp, 248 bp, 252 bp, 267 bp, 270 bp and null. In the foreign collection of varieties − Gli-B1a, Gli-B1b, Gli-B1c, Gli-B1d, Gli-B1e, Gli-B1f, Gli-B1g, Gli-B1h, Gli-B1i, Gli-B1j, Gli-B1k, Gli -B1l, Gli-B1m, Gli-B1n, Gli-B1o, Gli-B1p, Gli-B1q, Gli-B1r, Gli-B1s and 213 bp, 216 bp, 237 bp, 246 bp, 248 bp, 250 bp, 252 bp, 270 bp, 285 bp and null. Nucleotide sequence analysis in the NCBI database showed the presence of a number of other alleles of the Taglgap microsatellite not only in bread wheat but also in some species of the Triticum L. and Aegilops L. genus. Conclusions. The detected polymorphism correlates with the polymorphism of allelic variants of gliadins of Gli-B1 locus and makes it possible to identify Gli-B1a, Gli-B1d, Gli-B1h and Gli-B1l allelic variants, and for Ukrainian varieties with high probability also Gli-B1b allelic variant. However, this marker does not allow identifying Gli-B1c, which is important for selection.

Identifikasi Marka Polimorfik untuk Pemuliaan Padi Toleran Defisiensi Fosfor

<p>Information on polymorphisms<br />among rice parents are very important in rice<br />breeding for tolerance to phosphorus defficiency. A study<br />was conducted at the Molecular Biology Laboratory,<br />Indonesian Center Agricultural Biotechnology and Genetic<br />Resources (ICABIOGRAD) from October 2006 to July 2007 to<br />identify polymorphism markers from 6 rice genotypes. The<br />rice genotypes, i.e., Dodokan, Situ Bagendit, Batur, Kasalath,<br />NIL-C443, dan K36-5-1-1 were analyzed for polymorphisms<br />using 496 SSR markers, which cover the rice genomes.<br />Seven of the 496 markers were used as foreground and<br />recombinant selection markers, and the rests (489 markers)<br />were used as background selection markers. PCR amplifications<br />were separated on a 5% polyacrylamide gel and<br />colored by the silver staining method. Three different markers<br />among the seven foreground and recombinant selection<br />markers were selected from each crossing, which are<br />tightly linked with Pup1 gene and have a distance less than 5<br />cM. These markers are Dodokan vs Kasalath (RM277, SSR3,<br />RM519), Dodokan vs NIL-C443 (RM277, SSR3, RM519),<br />Dodokan vs K36-5-1-1 (RM277, SSR3, RM519), Situ Bagendit<br />vs Kasalath (RM28102, SSR3, RM519), Situ Bagendit vs NILC443<br />(RM28102, SSR3, RM519), Situ Bagendit vs K36-5-1-1<br />(RM511, SSR3, RM519), Batur vs Kasalath (RM277, RM1261,<br />RM519), Batur vs NIL-C443 (RM277, RM1261, RM519), and<br />Batur vs K36-5-1-1 (RM28102, SSR3). Variations in background<br />selection primers were found in each chromosome<br />and in each parent combinations. Primers on chromosome<br />4, 5, and 12 showed the lowest polymorphisms; more<br />primers are needed for these chromosomes.</p>

Changes in the number and size of nucleoli of Chara vulgaris L. antheridial filament cells during the period preceding light-induced re-initiation of cell divisions following a mitodepressive effect of darkness

The changes in number and size of nucleoli of <em>Chara vulgaris</em> antheridial filament cells were monitored with the use of Howell and Black's silver staining method. After a 3-day mitodepressive treatment with darkness the cells were exposed to light which reactivated mitotic activity after 18-20 hours. Eight-celled antheridial filaments were observed. In the period preceding light-induced re-initiation of mitoses a gradual reconstruction of the number and size of nucleoli characteristic of control, as well as their total area per nucleus appeared. The obtained results indicate that one of the important conditions for a cell to be able to divide is accumulation of nucleolus components characteristic of a given developmental stage and this controls nucleologenesis of the subsequent cell cycle.