Tumoral CD105 is a novel independent prognostic marker for prognosis in clear-cell renal cell carcinoma.

BACKGROUND: Angiogenesis is essential for tumour growth and metastasis. There are conflicting reports as to whether microvessel density (MVD) using the endothelial marker CD105 (cluster of differentiation molecule 105) in clear-cell renal cell carcinomas (ccRCC) is associated with prognosis. Recently, CD105 has been described as a RCC cancer stem cell marker. METHODS: A total of 102 ccRCC were analysed. Representative tumour sections were stained for CD105. Vascularity (endothelial CD105) was quantified by MVD. The immunohistochemistry analysis detected positive (if present) or negative (if absent) CD105 tumoral staining. This retrospective population-based study was evaluated using Kaplan-Meier method, t-test and Cox proportional hazard model. RESULTS: We found that the expression of endothelial CD105 (MVD) negatively correlated with nuclear grade (P<0.001), tumour stage (P<0.001) and Leibovitch score (P<0.001), whereas the expression of tumoral CD105 positively correlated with these three clinicopathological factors (P<0.001). In multivariate analysis, tumoral CD105 was found to be an independent predictor of poor overall survival (P=0.002). CONCLUSIONS: We have shown for the first time that tumoral CD105 is an independent predictive marker for death risk and unfavourable prognosis in patients with ccRCC after curative resection.

Virulence profiles and antibiotic susceptibility patterns of Klebsiella pneumoniae strains isolated from different clinical specimens.

AIM OF THE STUDY: To detect virulence factors in 54 Klebsiella pneumoniae isolates from different clinical specimens: urine (26), blood (11), pus (11), lung (four), cerebrospinal fluid (one) and ascitic fluid (one). MATERIAL AND METHODS: PCR was used to investigate virulence genes encoding adhesins (fimH-1, mrkD, kpn, ycfM), siderophores (entB: enterobactin, iutA: aerobactin, irp-1, irp-2, ybtS, fyuA: yersiniabactin, iroN: catechols receptor), protectines or invasins (rmpA, magA, traT) and toxins (hlyA, cnf-1). The serum resistance, capsule and hypermucoviscosity, and ability to form biofilm and produce siderophores were sought by phenotypic assays. The in vivo virulence was assessed in mice infected by intraperitoneal way. Antimicrobial susceptibility was tested by diffusion method. RESULTS: The most common virulence genes were fimH-1 (100%), mrkD (96.3%), ycfM (96.3%), and entB (100%). kpn and yersiniabactin genes were found at medium rates of 63% and 46.3% and at lower prevalence, were genes traT (1.8%), iroN (3.7%), iutA (5.5%) and rmpA (3.7%). magA, hlyA and cnf-1 genes were not detected. The capsule, serum resistance, biofilm formation, mannose-sensitive or -resistant haemagglutination and hypermucoviscosity were observed in 100%, 92.6%, 88.8%, 94.4%, 68.5% and 9.2% of isolates, respectively. The prevalence of siderophores was consistent with that of genotypic detection. The LD50 in mice was very low (<10(2) CFUs) for isolates with the most virulence factors. A rate of 74.1% of isolates showed a multidrug resistance (MDR) pattern. CONCLUSIONS: The distribution of virulence profiles according to the clinical origin suggests a role of enterobactin in urinary infections and yersiniabactin in the invasiveness. The fimbriae F1 and F3, capsule, enterobactin, serum resistance and biofilm formation, were commonly found in isolates, they seem to be at the basis of classic pathogenicity of K. pneumoniae. The invasiveness enhancers, aerobactin, yersiniabactin, catechols receptor, mucoid factor and hypermucoviscosity, detected concomitantly in some isolates, constitute a threat for vulnerable populations, even more if they are in combination with antibiotic resistance.

CTX-M-15 extended-spectrum beta-lactamases in Enterobacteriaceae in the intensive care unit of Tlemcen Hospital, Algeria.

The aim of this study was to detect extended-spectrum beta-lactamases (ESBL) in Enterobacteriaceae isolates in the intensive care unit (ICU) of Tlemcen hospital in north-western Algeria. Antimicrobial susceptibility testing, molecular typing, characterization of ESBL-encoding genes and the genetic environment, conjugation experiments and plasmid analysis were carried out. In all, 28 Enterobacteriaceae isolates were isolated from specimens recovered from patients in the ICU and 2 from surfaces of the unit. Of these, 11 isolates (4 Escherichia coli, 5 Klebsiella pneumoniae and 2 Enterobacter cloacae) produced ESBL of the CT-X-M-15 type. Molecular typing of the isolates showed the clonal nature of 4 K. pneumoniae isolates. The bla(CTXM-15) gene was genetically linked to insertion sequence lSEcp1B and was transferable by conjugation from 3 isolates. Regular monitoring of resistance mechanisms, the establishment of a prevention strategy, and more rational and appropriate use of antibiotics are needed.

CTX-M-15 extended-spectrum beta-lactamases in Enterobacteriaceae in the intensive care unit of Tlemcen hospital, Algeria

The aim of this study was to detect extended-spectrum beta-lactamases [ESBL] in Enterobacteriaceae isolates in the intensive care unit [ICU] of Tlemcen hospital in north-western Algeria. Antimicrobial susceptibility testing, molecular typing, characterization of ESBL-encoding genes and the genetic environment, conjugation experiments and plasmid analysis were carried out. In all, 28 Enterobacteriaceae isolates were isolated from specimens recovered from patients in the ICU and 2 from surfaces of the unit. Of these, 11 isolates [4 Escherichia coti, 5 Klebsiellapneumoniae and 2 Enterobacter cloacae] produced ESBL of the CT-X-M-15 type. Molecular typing of the isolates showed the clonal nature of 4 K, pneumonlae isolates. The bla[CTMX-15] gene was genetically linked to insertion sequence ISEcp1B and was transferable by conjugation from 3 isolates. Regular monitoring of resistance mechanisms, the establishment of a prevention strategy, and more rational and appropriate use of antibiotics are needed

Interleukin 10 promoter region polymorphisms in inflammatory bowel disease in Tunisian population.

OBJECTIVE: To test whether IL-10 promoter region polymorphisms are associated with susceptibility to inflammatory bowel disease, we examined the contribution of interleukin- 10 (IL-10) gene polymorphisms to Crohn's disease (CD) and Ulcerative colitis disease (UC) occurrence and also to CD phenotype. MATERIELS AND METHODS: SNPs at positions -627 (C > A) and -1117 (G > A) in the IL-10 promoter were determined in a sample of 105 Tunisian patients with IBD (75 CD and 30 UC) and 90 matched healthy controls. RESULTS: The 627 CA genotype is associated with ileal location (p = 0.015) and with stricturing (p = 510-3) and penetrating (p = 310-3) presentation of CD. An additive effect between IL10 variants and CARD15 3020 insC mutation (p = 0,006) on severe forms of CD was shown. CONCLUSIONS: In Tunisian population, the 3020insC insertion in NOD2/CARD15 gene is a marker of susceptibility to CD, while the A allele at position -627 in the IL-10 promoter increases the risk of CD ileal location and severe disease presentation. A genetic epistasis between IL-10 gene polymorphisms and CARD15/NOD2 gene mutation was suggested.

3020insC insertion in NOD2/CARD15 gene, a prevalent variant associated with anti-Saccharomyces cerevisiae antibodies and ileal location of Crohn's disease in Tunisian population.

OBJECTIVE: Our aim is to investigate the relation between CARD15 3020insC mutation, anti-Saccharomyces cerevisiae antibodies (ASCA) and disease phenotype, in Tunisian inflammatory bowel disease (IBD) patients. MATERIALS: A hundred Tunisian patients with IBD (75 Crohn's disease CD and 25 ulcerative colitis UC) and 60 matched healthy controls were studied. METHODS: CARD15 mutation was analysed by using an allele-specific polymerase chain reaction and sequencing. Assessment of ASCA in serum was performed by ELISA. RESULTS: The frequency of the mutation was significantly higher in Crohn's disease than in control (p = 0,0005; OR = 20.45; CI 95% = 2.86-413.85) and did not differ statistically in UC group (p = 0, 05) from control. ASCAs were present in 60% of CD and 20, 8% of UC. CONCLUSION: This study suggests that in northern Tunisian population, 3020insC mutation in NOD2/CARD15 gene is a prevalent mutation leading to the typical Crohn's disease including ileal location, stricturing and penetrating clinical types and ASCA expression.

Prevalence and characterization of extended-spectrum beta-lactamases in Klebsiella pneumoniae in Algiers hospitals (Algeria).

AIM OF THE STUDY: To determine the prevalence and the diversity of extended-spectrum beta-lactamases (ESBLs) in 196 Klebsiella pneumoniae clinical isolates collected from three hospitals in Algiers. MATERIALS AND METHODS: Antibiograms were done on Mueller-Hinton agar plates with the disc-diffusion method and MICs were determined by agar-dilution method. Mating experiments were performed in agar medium. Plasmid DNA was extracted by the alcalin-lysis method. Total DNA was extracted with a Qiagen mini kit and screened for bla(TEM) and bla(CTX-M) genes by PCR. Linkage of bla(CTX-M) genes with insertion sequence ISEcp1B and class 1 integrons was investigated by PCR. PCR products were sequenced by the Sanger method. The epidemiological relationships between ESBL-producing K. pneumoniae isolates were analyzed by ERIC-PCR. RESULTS: Thirty-nine (19.9%) isolates were found to produce ESBLs belonging to CTX-M-1 group and TEM penicillinases (CTX-M-3, CTX-M-15 and TEM-1). ERIC-PCR analysis showed that the isolates are genetically unrelated. The bla(TEM) and bla(CTX-M) genes as well as aminoglycosides and sulfonamides resistance determinants were found located in self-transferable plasmids of approximately 85 kb. The class 1 integrons and the insertion sequence ISEcp1B were present in the isolates and in their transconjugants. ISEcp1B was found genetically linked to the bla(CTX-M) genes and located 127bp upstream, with the presence of the V and W sequences. CONCLUSION: The study revealed a high rate of ESBL-producing K. pneumoniae in Algerian hospitals, resulting from horizontal dissemination of mobile bla(CTX-M) genes.

Detection of cytokeratin 19 mRNA and CYFRA 21-1 (cytokeratin 19 fragments) in blood of Tunisian women with breast cancer.

Cytokeratin 19 (CK19) is an acidic protein of 40 kDa that is part of the cytoskeleton of epithelial cells. It is highly expressed by all epithelial cells and represents a useful indicator of epithelial differentiation. The soluble fragment of CK19 (CYFRA 21-1) can be a useful circulating tumor marker and can be detected in the serum of cancer patients. The development of metastasis in patients with cancer of epithelial origin is due to the migration of tumor cells from the original tumor to distant organs. In order to detect micrometastasis in patients with breast cancer, we evaluated and compared CK19 gene expression using RT-PCR in blood samples collected from 80 healthy women and 80 patients with localized or metastatic breast cancer. The concentration of the soluble CK19 fragment CYFRA 21-1 was measured in serum of all study subjects by radioimmunoassay employing specific monoclonal antibodies. The relationship between the expression of this molecular marker and clinical stage, tumor differentiation and CK19 mRNA transcripts was investigated. We found that CK19 mRNA expression in blood (as a direct index of the presence of circulating tumor cells) was not correlated with CYFRA 21-1.

Prevalence of beta-lactams resistance among Escherichia coli clinical isolates from a hospital in Algiers.

A high prevalence of beta-lactams resistance among Enterobacteriaceae have been reported worldwide; however, there are not sufficient data on this issue in Algeria. beta-Lactams susceptibility of 203 Escherichia coli clinical isolates was determined by agar diffusion method, and production of extended-spectrum beta-lactamases (ESBL) was screened by double-disk synergy test. This analysis showed five well-defined phenotypes: 1) 62 isolates (30.5%) were susceptible to all beta-lactams; 2) 135 isolates (66.5%) presented a broad-spectrum beta-lactamases phenotype (BSBL); 3) three isolates (1.5%) were defined as producing ESBLs; 4) two isolates (1%) were AmpC cephalosporinase producers; and 5) one isolate (0.5%) presented a phenotype of cell-decreased permeability to beta-lactams. Isoelectric focusing revealed beta-lactamases with isolectric points of 5.4 or 7.6 for isolates with BSBL phenotype; approximately 9.0 for two ESBL isolates; 5.4, 7.6 and approximately 9.0 for the remaining ESBL isolate; and 5.4 and approximately 9.0 for the AmpC isolates. The cefotaxime hydrolysis corresponds to the basic bands with an isoelectric point of approximately 9.0. Conjugation assay showed transfer of penicillinase and AmpC resistance phenotypes and their corresponding beta-lactamases to recipient E. coli BM21 in association with plasmids of 71.4 kb for the AmpC isolates and from 40-56 kb for penicillinase isolates. This result showed that the AmpC phenotype is plasmid mediated. ESBL isolates were found not to transfer their resistance through conjugation experiment. Polymerase chain reaction (PCR) experiments using primers specific to blaTEM, blaAmpC and blaCTX-M genes showed specific amplification with blaCTX-M primer for two ESBL isolates; blaTEM and blaCTX-M for the remaining ESBL isolate; and blaTEM and blaAmpC for the AmpC isolates and their corresponding transconjugants. The study showed a high rate of isolates producing penicillinase, and low frequencies of AmpC and ESBL phenotypes. AmpC beta-lactamases were plasmid mediated, and ESBLs belong to the CTM-M type.

Prevalence of β - lactams resistance among Escherichia coli clinical isolates from a hospital in Algiers

La prevalencia de la resistencia a los betalactámicos entre las enterobacterias es alta en todo el mundo, pero en Argelia no se dispone desuficientes datos. Se determinó la sensibilidad a los betalactámicos de 203 cepas clínicas de Escherichia coli mediante difusión en agar y seanalizó la producción de betalactamasas de espectro extendido (BLEE) mediante la técnica de la sinergia del doble disco. Este análisis mostrócinco fenotipos bien definidos: 1) 62 cepas (30,5%) fueron sensibles a todos los betalactámicos; 2) 135 cepas (66,5%) presentaron unfenotipo caracterizado por betalactamasas de amplio espectro (BLEA); 3) 3 cepas (1,5%) se definieron como productoras de BLEE; 4) 2 cepas(1%) fueron productoras de cefalosporinasa tipo AmpC; y 5) una (0,5%) presentó un fenotipo de disminución de la permeabilidad celulara los betalactámicos. La determinación del punto isoeléctrico mostró betalactamasas con puntos isoeléctricos de 5,4 o 7,6 para las cepascon fenotipo BLEA; ∼9,0 para 2 cepas productoras de BLEE; 5,4, 7,6 y ∼9,0 para la tercera cepa productora de BLEE; y 5,4 y ∼9,0 para lascepas productoras de cefalosporinasa AmpC. La hidrólisis de cefotaxima se corresponde con las bandas básicas con un punto isoeléctricode ∼9,0. El ensayo de conjugación mostró una transferencia de los fenotipos de resistencia de penicilinasas y cefalosporinasa AmpC y suscorrespondientes betalactamasas a E. coli BM21 en asociación con plásmidos de 71,4 kb para las cepas productoras de cefalosporinasaAmpC y de 40-56 kb para las productoras de penicilinasas. Este resultado mostró que el fenotipo productor de cefalosporinasa AmpC estámediado por plásmidos. Las cepas productoras de BLEE no transfirieron su resistencia mediante el ensayo de conjugación. La reacción encadena de la polimerasa utilizando primers específicos para los genes blaTEM, blaAmpC y blaCTX-M mostró una amplificación específica con elprimer para blaCTX-M en dos cepas productoras de BLEE; los primers para blaTEM y blaCTX-M para la tercera cepa productora de BLEE; y losprimers para blaTEM y blaAmpC para las cepas productoras de cefalosporinasa AmpC y sus cepas transconjugantes correspondientes. El estudiomostró una alta tasa de cepas productoras de penicilinasas y una baja frecuencia de fenotipos productores de AmpC y BLEE. Las betalactamasasproductoras de cefalosporinasa AmpC estaban mediadas por plásmidos y las BLEE pertenecieron al tipo CTM-M

A high prevalence of β-lactams resistance among Enterobacteriaceae have been reported worldwide; however, there are not sufficient dataon this issue in Algeria. β-Lactams susceptibility of 203 Escherichia coli clinical isolates was determined by agar diffusion method, and productionof extended-spectrum β-lactamases (ESBL) was screened by double-disk synergy test. This analysis showed five well-defined phenotypes:1) 62 isolates (30.5%) were susceptible to all β-lactams; 2) 135 isolates (66.5%) presented a broad-spectrum β-lactamases phenotype(BSBL); 3) three isolates (1.5%) were defined as producing ESBLs; 4) two isolates (1%) were AmpC cephalosporinase producers; and 5) one isolate(0.5%) presented a phenotype of cell-decreased permeability to β-lactams. Isoelectric focusing revealed β-lactamases with isolectric pointsof 5.4 or 7.6 for isolates with BSBL phenotype; ∼9.0 for two ESBL isolates; 5.4, 7.6 and ∼9.0 for the remaining ESBL isolate; and 5.4 and ∼9.0for the AmpC isolates. The cefotaxime hydrolysis corresponds to the basic bands with an isoelectric point of ∼9.0. Conjugation assay showedtransfer of penicillinase and AmpC resistance phenotypes and their corresponding β-lactamases to recipient E. coli BM21 in association withplasmids of 71.4 kb for the AmpC isolates and from 40–56 kb for penicillinase isolates. This result showed that the AmpC phenotype is plasmidmediated. ESBL isolates were found not to transfer their resistance through conjugation experiment. Polymerase chain reaction (PCR) experimentsusing primers specific to blaTEM , blaAmpC and blaCTX-M genes showed specific amplification with blaCTX-M primer for two ESBL isolates;blaTEM and blaCTX-M for the remaining ESBL isolate; and blaTEM and blaAmpC for the AmpC isolates and their corresponding transconjugants. Thestudy showed a high rate of isolates producing penicillinase, and low frequencies of AmpC and ESBL phenotypes. AmpC β-lactamases wereplasmid mediated, and ESBLs belong to the CTM-M type