First description of mutational analysis of MLH1, MSH2 and MSH6 in Algerian families with suspected Lynch syndrome.

Hereditary non-polyposis colorectal cancer (HNPCC) is an autosomal dominant disorder characterized by the early onset of colorectal cancer (CRC) linked to germline defects in Mismatch Repair (MMR) genes. We present here, the first molecular study of the correlation between CRC and mutations occurring in these genes performed in twenty-one unrelated Algerian families. The presence of germline mutations in MMR genes, MLH1, MSH2 and MSH6 genes was tested by sequencing all exons plus adjacent intronic sequences and Multiplex ligand-dependent probe amplification (MLPA) for testing large genomic rearrangements. Pathogenic mutations were identified in 20 % of families with clinical suspicion on HNPCC. Two novel variants described for the first time in Algerian families were identified in MLH1, c.881_884delTCAGinsCATTCCT and a large deletion in MSH6 gene from a young onset of CRC. Moreover, the variants of MSH2 gene: c.942+3A>T, c.1030C>T, the most described ones, were also detected in Algerian families. Furthermore, the families HNPCC caused by MSH6 germline mutation may show an age of onset that is comparable to this of patients with MLH1 and MSH2 mutations. In this study, we confirmed that MSH2, MLH1, and MSH6 contribute to CRC susceptibility. This work represents the implementation of a diagnostic algorithm for the identification of Lynch syndrome patients in Algerian families.

Clonal relationship between human and avian ciprofloxacin-resistant Escherichia coli isolates in North-Eastern Algeria.

The objectives of this study were to determine rates, patterns, and mechanisms of antibiotic resistance, and to assess connections between chicken commensal, human commensal, and pathogenic ciprofloxacin-resistant Escherichia coli isolates. All E. coli isolates collected from chickens, their farmers, and patients in the Constantine region (North-east Algeria) were analyzed for bla and plasmid-mediated quinolone resistance (PMQR) gene contents, phylogroups, Rep-PCR profiles, and multilocus sequence types. A high prevalence of resistance to fluoroquinolones (51.4 % to ciprofloxacin) was recorded in avian isolates. Of these, 22.2 % carried the aac(6')-Ib-cr gene, whereas lower resistance levels to these antibiotics were recorded in chicken farmers' isolates. None of the commensal isolates harbored the qnr, qepA, or oqxAB genes. One human pathogenic isolate was ertapenem-resistant and harbored the bla OXA-48 gene, 84 showed an extended-spectrum ß-lactamase phenotype, with bla CTX-M-15 gene prevalent in 87.2 % of them. Seventy isolates were resistant to fluoroquinolones, with aac(6')-Ib-cr present in 72.8 %, qnrB in 5.7 %, and qnrS in 10 %. Three Rep-PCR profiles were common to chicken commensal and human pathogenic isolates (phylogroups D and B1; ST21, ST48, and ST471 respectively); one was found in both chicken and chicken-farmer commensal strains (D; ST108), while another profile was identified in a chicken-farmer commensal strain and a human pathogenic one (B1; ST19). These findings suggest clonal and epidemiologic links between chicken and human ciprofloxacin-resistant E. coli isolates and the important role that poultry may play in the epidemiology of human E. coli infections in the Constantine region.

First description of OXA-48-producing Escherichia coli and the pandemic clone ST131 from patients hospitalised at a military hospital in Algeria.

The aim of the study was to assess the frequency and diversity of carbapenemases and extended-spectrum ß-lactamases (ESBL) produced by Escherichia coli isolates from patients hospitalised in the Regional Military Hospital of Constantine (Algeria). E. coli isolates were collected over a 2-year period from patients presenting E. coli infections. Strains with reduced susceptibility to ertapenem and/or positive for ESBL were characterised with regard to antibiotic resistance, bla genes, phylogenetic groups, O25 serotyping, quinolone resistance, repetitive sequence-based polymerase chain reaction (rep-PCR) profiles and multi-locus sequence typing (MLST). Of the 448 isolated E. coli, 94 (20.9 %) were multidrug-resistant. One of them (1.1 %) produced a bla OXA-48 and was identified as a B1 ST5 strain. The transposon bearing this gene was Tn1999.2. This strain was isolated from a patient coming from a border province with Tunisia, where this carbapenemase is endemic. In addition, 84 (18.8 %) isolates among them produced an ESBL with predominance (97.6 %) of bla CTX-M-15, which was coupled with qnr genes in 10.9 %. ESBL-producing strains were mainly detected in phylogroups D and A. They displayed 20 rep-PCR profiles and all the clonally related isolates were of the same sequence type (ST). Ten strains (9.4 %) belonged to the pandemic clone ST131. This study describes for the first time the presence of OXA-48-producing E. coli and the emergence of the intercontinental ST131 bla CTX-15-producing E. coli strains in Algeria.

Pre-harvest treatments with fungicides and post-harvest dips in sodium bicarbonate to control postharvest decay in stone fruit.

The objective of this study was to evaluate the effectiveness of different commercial formulations of fungicides containing one or more active ingredients in controlling postharvest decay of Thyrinthos and Boccuccia apricots, Red top peaches and Caldesi nectarines. Field treatments consisted of two sprays with cupric compounds, at the end of leaf fall and before bud swelling, one with sulfur compound, at fruit about half final size stage, and one with one of the following commercial formulations at the label suggested rates, one week before harvest: Teldor (fenexamid 50%; Bayer Crop Protection), Folicur (Tebuconazole 4.35%; Bayer Crop Protection), Signum (boscalid 26.7%, pyraclostrobin 6.7%; Basf Crop Protection), Score (difenoconazole 23.23%, Syngenta Crop Protection) and Switch (cyprodinil 37.5%, fludioxonil 25%, Syngenta Crop Protection). After harvest the fruit were stored for 1 week at 6 degrees C and 90% RH followed by 1 week at 20 degrees C and 60% RH to simulate retail conditions, or placed directly at 20 degrees C. All formulations significantly reduced decay in all cultivars. Switch, Signum and Folicur were the most active, while Score was slightly less effective. Teldor activity was low, especially in Thyrintos apricots, where the percentage of rotten fruit was slightly lower than in control fruit. Brown rot was the most representative disease, but in apricots a high percentage of fruit was affected by blue mold and grey mold. Rhizopus rot generally developed as a secondary disease on fruit previously affected by other pathogens and was more frequent in control and Teldor treated fruit. Preharvest sprays with Signum 3 days before harvest reduced postharvest decay after 1 week storage at 20 degrees C in Glo haven peaches and Venus nectarine harvested at advanced stage of maturity. Combining pre-harvest sprays with Signum and a 2-min postharvest dip in 2% sodium bicarbonate at 20 degrees C further reduced decay. In Sothern regions of Italy, the use of synthetic fungicides only immediately before harvest in years when the weather conditions are not favorable to brown rot and other pathogens inducing postharvest decay, combined with a postharvest treatment with sodium bicarbonate could be a feasible integrated approach to reduce the risk of selection of resistant strains of fungi to synthetic fungicides while controlling effectively postharvest decay.

Immunostimulating activity of the hot water-soluble polysaccharide extracts of Anacyclus pyrethrum, Alpinia galanga and Citrullus colocynthis.

Hot water polysaccharide extracts of Anacyclus pyrethrum (L.) Link. (family Compositae) Citrullus colocynthis (L.) Schrad. (family Cucurbitaceae) and Alpinia galanga (L.) Willd. (family Zingiberaceae) were tested for their immunostimulating activity in mice. The fractions from Anacyclus pyrethrum and Alpinia galanga showed a marked stimulating effect on the reticulo-endothelial system (RES) and increased the number of peritoneal exudate cells (PEC), and spleen cells of mice. In this case, the optimum doses were 50 and 25 mg/kg for the two fractions, respectively. On the other hand, the polysaccharide extracts of both Anacyclus pyrethrum and Alpinia galanga markedly enhanced the proliferation of the murine spleen cells in vitro using two tests (in vitro and in vivo effect). The results of the in vivo effect at a doses of 50 and 25 mg/kg, showed a stimulation index better than obtained with the in vitro effect at 50 and 25 microg/ml for Anacyclus pyrethrum and Alpinia galanga, respectively. While the extract of Citrullus colocynthis showed much weaker and variable immunostimulating activity.

A novel C-->A transversion within the distal CCAAT motif of the Ggamma-globin gene in the Algerian Ggammabeta+-hereditary persistence of fetal hemoglobin.

Hereditary persistence of fetal hemoglobin (HPFH) is a group of genetically heterogeneous conditions characterized by the continued expression of fetal hemoglobin in adulthood. These constitute natural models for understanding the mechanism(s) of the hemoglobin switch. Many large deletions in the beta-globin gene cluster and point mutations in one of the fetal globin gene promoters have been described before. In this study we describe a novel C-->A transversion (-114) in the distal CCAAT box of the Ggamma-globin gene promoter associated with the Ggammabeta+-HPFH phenotype in an Algerian family. Individuals heterozygous for this mutation exhibit moderate raise in Hb F levels (0.6-3.5%). Much higher Hb F levels (3.8-11.2%) are observed when a beta(o)-thalassemia allele is present in trans to the hereditary persistence of fetal hemoglobin allele. This novel Algerian HPFH mutation further stresses the importance of the distal CCAAT box in the postnatal regulation of gamma-globin gene expression.

Solubilization of plasma membranes in anionic, non-ionic and zwitterionic surfactants for iso-dalt analysis: a critical evaluation.

A critical study has been made of the solubilizing properties of anionic, nonionic and zwitterionic surfactants to be used in the first dimension of two-dimensional isoelectric focusing-sodium dodecyl sulphate (IF-SDS) maps. Excess of SDS is a powerful solubilizing agent, but prevents proteins from entering the IF gel. Nonidet P-40 (NP-40)-urea mixtures are mediocre solubilizing agents, but are compatible with the IF dimension. Zwitterionic detergents (zwittergents) appear to exhibit a well balanced solubilizing power and are able to generate sharp two-dimensional maps, with round spots and minimal vertical and horizontal streaking. SB3-12 and SB3-14 appear to have the best solubilizing properties; shorter surfactants (SB3-8) exhibit a poor solubilization efficiency, while longer detergents (SB3-16) bind too strongly to hydrophobic regions in proteins. The random performance of non-ionic detergents has been attributed to their ability to form mixed micelles with the carrier ampholytes used in the IF step: depending on the relative ratio of NP-40 to Ampholines, different types of micelles would be formed, which, when reaching appropriate stoichiometries and charge densities, would mimic the behaviour of natural zwittergents. Acceptable two-dimensional maps can be obtained when the sample is lysed in limiting amounts of SDS (SDS:protein = 1:3), suggesting that excess of free SDS in solution is deleterious to the IF process.