Intragenomic and intraspecific heterogeneity of the 16S rRNA gene in seven bacterial species from the respiratory tract of cystic fibrosis patients assessed by PCR-Temporal Temperature Gel Electrophoresis.

16S rRNA gene-based cultivation-independent methods are increasingly used to study the diversity of microbiota during health and disease. One bias of these methods is the variability of 16S rRNA gene that may exist among strains of a same species (intraspecific heterogeneity) or between rrs copies in a genome (intragenomic heterogeneity). We evaluated the level of intraspecific and intragenomic 16S rDNA variability in seven species frequently encountered in respiratory tract samples in cystic fibrosis (CF). A total of 179 strains were subjected to V3 region 16S rDNA PCR-TTGE. Using this easy-to-perform and rapid method, different levels of V3 region rrs heterogeneity were demonstrated. No intraspecific and intragenomic rrs heterogeneity was demonstrated for Moraxella catarrhalis (n=16), Pseudomonas aeruginosa (n=31) and Streptococcus pneumoniae (n=14) showing a single PCR-TTGE band characteristic of the species. Low level of intraspecific heterogeneity was observed for Staphylococcus aureus (n=30), Stenotrophomonas maltophilia (n=29) and Achromobacter xylosoxidans (n=28), and 17%, 38% and 96% of these strains showed intragenomic heterogeneity (two to four different rrs copies), respectively. Haemophilus influenzae (n=31) displayed the higher level of intraspecific variability with 23 different PCR-TTGE patterns and 61% of the strains showed intragenomic rrs heterogeneity (two to four different rrs copies). Although only one hypervariable region of the 16S rRNA gene was explored, intraspecific and intragenomic rrs heterogeneity was frequently observed in this study and should be taken into consideration for a better interpretation of 16S rRNA gene-based diversity profiles in denaturing gels and to avoid any overestimation of the respiratory microbiota diversity in CF.

Drowning associated pneumonia: a descriptive cohort.

PURPOSE: Pneumonia is the most common infectious complication of drowning. Pneumonia is potentially life threatening and should be treated by effective antibiotic therapy. However the risk factors, microbiological causes, diagnostic approach and appropriate therapy for pneumonia associated with drowning are not well described. The microbiological ecology of the body of water where immersion occurred could be of import. The aim of this study was to report on microorganisms involved in pneumonia associated with drowning and out of hospital cardiac arrest after successful cardiopulmonary resuscitation. Additionally, we retrieved and undertook microbiological analysis on samples of water from our local river. METHODS: This retrospective study included all patients having suffered an out of hospital cardiac arrest due to drowning and admitted to our tertiary care academic hospital between 2002 and 2010. Data concerning bacteriological lung samples (tracheal aspirate or bronchoalveolar lavage) at admission were reported and compared to bacteriological samples obtained from our local river (the river Seine). RESULTS: A total of thirty-seven patients were included in the study. Lung samples were obtained for twenty-one of these patients. Lung samples were positive in nineteen cases, with a high frequency of multi-drug resistant bacteria. Samples from the Seine River found microorganisms similar to those found in drowning associated pneumonia. CONCLUSIONS: Drowning associated pneumonia can be due to multi drug resistant bacteria. When treating drowning associated pneumonia, antibiotics should be effective against bacteria similar to those found in the body of water where immersion occurred.

Variación del Índice de Masa Corporal en niños tratados por sobrepeso y obesidad del Instituto Materno Infantil y Especialidades. 2003-2006 / Variation of the index of body mass in children treated by overweight and obesity of the maternal and specialties institute. 2003-2006

Se realiza un estudio retrospectivo y longitudinal en el Instituto Materno Infantil y Especialidades de Santiago de Los Caballeros, República Dominicana para conocer la variación del Índice de Masa Corporal (IMC), el tiempo de permanencia en el tratamiento, el rango de edad más afectado, de acuerdo al sexo, en 45 pacientes, 23 femeninos y 22 masculinos, con obesidad y sobrepeso, durante el período junio 2003 a mayo 2006. Se excluyeron del estudio a pacientes que no clasificaron como obesidad exógena. Resultados: Al inicio del tratamiento 100% de los pacientes se encontraba en rangos de IMC total correspondientes a Obesidad intensa, y por encima del 95 Percentil de IMC de acuerdo a las gráficas convencionales. El rango de 11 a 13.11 años presentó mayor número de hombres obesos, en tanto que el sexo femenino superó al masculino en el rango 2 a 8. En la fase post tratamiento se observó una reducción del IMC en el grupo total que descendió de 100 por encima del p 95 a solo 53.5% sobre este percentil. El tiempo de Permanencia en el tratamiento fluctuó entre 2 y 24 meses, con una Moda de 3 y una DS de 4.9. El IMC del grupo se redujo en un promedio de 14.8%. La Moda se situó en 11. La DS 5.8. Se concluye que el tratamiento para la obesidad benefició a todos los pacientes, en el corto plazo y medio plazo, recomendándose implementar estrategias para una adhesión al tratamiento a largo plazo

It is a retrospective and longitudinal study, realized in the Maternal Infantile and Specialties Institute, in Santiago, Dominican Republic, for to determine the Body mass Index (IMC) variation, permanency time, age and sex, i n 45 patients, 23 girls and 22 boys, with overweight and obesity, since June 2003 until May 2006. Patients without exogenous overweight and obesity were excluded to the study. Results: Previous to the treatment 100% of the patients were corresponding with intense obesity, over 95 percentile, prevailing boys over girls in range 11 - 13.11 years, and girls over boys in range 2-8 years. Posterior to the treatment 53% of patients remained over 95 percentile, whereas the 47% place under 95 IMC percentile. The permanence time in the treatment fluctuated between 2-24 months. The modal value was 3, and the Standard Deviation (SD) 4.9. The BMI of the group was reduced in 14.8%, the modal value was 11, and SD 5.8. In conclusion: the obesity treatment beneficed all patients, in short and medial term. Recommendations is to work strategy for more adhesion at long term (AU)

Functional characterization of human CD34+ cells that express low or high levels of the membrane antigen CD111 (nectin 1).

Nectins are recently described adhesion molecules that are widely expressed on many tissues, including the hematopoietic tissue. Nectin 1 (CD111) is expressed on a higher proportion of mobilized peripheral blood (mPB) than cord blood (CB) CD34+ cells, and of CD34+/CD38+ cells when compared with CD34+/CD38- cells. We studied functional properties of human CB and mPB CD34+ cells that express low or high levels of CD111. CD34+/CD111(dim) cells contain a higher proportion of cells in G0/G1 phase than CD34+/CD111(bright) cells. CD34+/CD111(bright) cells contain more erythroid progenitors: CFU-E, than their counterparts, which on the opposite contain more HPP-CFC. Limiting dilution analyses demonstrate a higher frequency of immature progenitors: cobblestone-area colony-forming cells, CD34+/CD111(dim) than in CD34+/CD111(bright) cells. In vitro differentiation assays demonstrate a higher frequency of B-, T- and dendritic-cell precursors, but less NK-cell precursors in CD34+/CD111(dim) cells. Evaluation of engraftment in NOD-SCID mice shows that SCID repopulating cells are more frequent among mPB CD34+/CD111(dim) cells. Liquid culture of CD34+/CD111(dim) cells with erythropoietin shows that CD111 expression increases simultaneously with CD36, following CD71 and before glycophorin A expression. In conclusion, immature human hematopoietic progenitors express low levels of CD111 on their surface. During erythroid differentiation CD34+ cells acquire higher levels of the CD111 antigen.

[Diagnostic strategy when confronted with a moderate and prolonged increase of transaminases]. / Démarche diagnostique devant une augmentation modérée et prolongée des transaminases.

TWO TYPES OF SITUATIONS: Measurement of transaminase serum activity is a common biological test. Although the etiological scope of acute and severe hyper-aminotransferase is codified and limited, that of prolonged and moderate hyper-aminotransferase is much broader. IN THE CASE OF PROLONGED AND MODERATE INCREASE IN TRANSAMINASE SERUM ACTIVITY: The discovery of this abnormality during systematic biological controls is a frequent situation, and its management is relatively well standardised. It requires a rigorous diagnostic strategy, which includes the search for consumption of alcohol, overweight, chronic hepatic disease of viral origin and the nature of the medicinal products ingested. FROM AN ETIOLOGICAL POINT OF VIEW: The most frequent causes of moderate and prolonged hyper-aminotransferase are alcohol abuse, overweight, non-insulin-dependent diabetes, dyslipaemia, viral hepatitis and medicinal products. However, less frequent hepatic or extra-hepatic causes must not be neglected.

A novel mechanism for thalassaemia intermedia.

Thalassaemia intermedia is a moderate form of thalassaemia resulting from various genetic defects. We report an undescribed mechanism leading to this condition: a somatic deletion of the beta-globin gene in the haemopoietic lineage of a heterozygous beta-thalassaemic patient. We did molecular studies and haemoglobin analysis of the patient and his parents. We found that the deletion gives rise to a mosaic of cells with either one or no functional beta-globin gene and it extends to a region of frequent loss of heterozygosity called LOH11A, which is located close to the beta-globin locus. Thus, loss of heterozygosity can be a cause of non-malignant genetic disease.

Forced expression of the Ikaros 6 isoform in human placental blood CD34(+) cells impairs their ability to differentiate toward the B-lymphoid lineage.

Studies in mice suggest that the Ikaros (Ik) gene encodes several isoforms and is a critical regulator of hematolymphoid differentiation. Little is known on the role of Ikaros in human stem cell differentiation. Herein, the biological consequences of the forced expression of Ikaros 6 (Ik6) in human placental blood CD34(+) progenitors are evaluated. Ik6 is one of the isoforms produced from the Ikaros premessenger RNA by alternative splicing and is thought to behave as a dominant negative isoform of the gene product because it lacks the DNA binding domain present in transcriptionally active isoforms. The results demonstrate that human cord blood CD34(+) cells that express high levels of Ik6 as a result of retrovirally mediated gene transfer have a reduced capacity to produce lymphoid B cells in 2 independent assays: (1) in vitro reinitiation of human hematopoiesis during coculture with the MS-5 murine stromal cell line and (2) xenotransplantation in nonobese diabetic-severe combined immunodeficient mice. These results suggest that Ikaros plays an important role in stem cell commitment in humans and that the balance between the different isoforms is a key element of this regulatory system; they support the hypothesis that posttranscriptional events can participate in the control of human hematopoietic differentiation.

Alteraciones digestivas frecuentes en niños con trastorno psicomotor / Frequents digestives alterations in children with psicomotrical trouble

La calidad de vida de los niños que padecen trastorno psicomotor puede estar directamente afectada por la presencia de diversas patologías que se suman a su enfermedad de base. Estas alteraciones pueden ser de carácter sensitivo motor, cognitivo y conductual. A nivel del tracto digestivo son frecuentes las patologías bucales de diverso orden, los trastornos deglutorios faríngeos y esofágicos, las alteraciones en el tono del cardias y del píloro, el vaciado gástrico lento, el reflujo gastroesofágico, la gastritis y el estreñimiento. Estas alteraciones, diversas en su expresión e intensidad, afectan en grado variable la conducta alimentaria y su calidad de vida total. El riesgo de padecer malnutrición es elevado en este grupo de pacientes. Por ello es importante la detección precoz de estos trastornos para aplicar a tiempo un tratamiento adecuado. La interacción de un equipo interdisciplinar y la aplicación de una terapia reeducativa es de suma trascendencia (AU)

Delta N89 beta-catenin induces precocious development, differentiation, and neoplasia in mammary gland.

To investigate the role of beta-catenin in mammary gland development and neoplasia, we expressed a stabilized, transcriptionally active form of beta-catenin lacking the NH(2)-terminal 89 amino acids (Delta N 89 beta-catenin) under the control of the mouse mammary tumor virus long terminal repeat. Our results show that Delta N 89 beta-catenin induces precocious lobuloalveolar development and differentiation in the mammary glands of both male and female mice. Virgin Delta N 89 beta-catenin mammary glands resemble those found in wild-type (wt) pregnant mice and inappropriately express cyclin D1 mRNA. In contrast to wt mammary glands, which resume a virgin appearance after cessation of lactation, transgenic mammary glands involute to a midpregnant status. All transgenic females develop multiple aggressive adenocarcinomas early in life. Surprisingly, the Delta N89 beta-catenin phenotype differs from those elicited by overexpression of Wnt genes in this gland. In particular, Delta N 89 beta-catenin has no effect on ductal side branching. This suggests that Wnt induction of ductal branching involves additional downstream effectors or modulators.

[Hepatotoxicity of medications]. / Du nouveau dans l'hépatotoxicité des médicaments.

A TIMELY TOPIC: Liver toxicity remains a common problem despite adequate information for physicians and drug watch programs. The number of recent publications reporting severe drug-induced liver disease emphasizes the need for prudence. ACUTE AND CHRONIC HEPATOTOXICITY: Cases of acute drug-induced liver disease have been described for nearly all drug classes: a few examples concern hepatitis subsequent to administration of fluoxetin, acarbose, riluzole, coumarin, or orlistat. Fulminant hepatitis is fortunately an exceptional event but has been described after administration of ketoprofene, nimesulid, and clarithyromycin. Chronic liver disease has also resulted from the use of mesalazine, minocyclin or fibrates. Nevirapin prescribed for HIV infection can cause severe liver disease. OTHER AGENTS: Certain herbal agents, such as chelidoin for example, can cause cholestasis. Certain excipients can also be toxic for the liver. Ecstasy appears to be a frequent cause of sometimes severe liver disease in younger subjects.

Tratamiento nutricional del niño con patología neurológica / Nutritional treatment in child with neurologycal patology

El riesgo de padecer malnutrición es muy alto en niños y adolescentes que padecen determinado tipo de patología neurológica (PN) por una diversidad de factores que en ellos concurren: trastornos de coordinación motora de la deglución en sus distintas fases, reflujo gastroesofágico, esofagitis, gastritis, alteración del reflejo gastrocólico, estreñimiento, anorexia, etc. Estos trastornos alteran la proporcionalidad deseable entre la cantidad de alimento aportado al paciente y las ingestas netas del mismo. Debido a ello se hace necesaria 1) la valoración periódica y sistemática del estado nutricional, adaptada a este tipo de pacientes, y a partir de los datos recabados 2) la toma de decisiones oportunas referidas a su alimentación: requerimientos, opciones para el soporte nutricional, vía de administración y 3) tratamiento de las complicaciones digestivas si las hubiere. Con ello se contribuirá a mejorar su calidad de vida y a disminuir el nivel de estrés que viven las personas encargadas de su cuidado y alimentación. Importa mucho abordar con acierto las particularidades en su comportamiento alimentario. Estas acciones, complejas en su abordaje, requieren un enfoque interdisciplinar. La intencionalidad de este trabajo es contribuir a estos objetivos (AU)

Valoración del estado nutricional en niños y adolescentes con trastorno psicomotor / Evaluation of the nutritional state in children and teen-agers with psychomotrical troubles

Si en cualquier paciente es importante la valoración de su estado nutricional, ésta se hace imprescindible en niños y adolescentes con Trastorno Psicomotor. Se sabe que un déficit de energía, proteínas y micronutrientes, interfiere con el crecimiento, desarrollo y función neuromuscular de niños y adolescentes sanos. En los que padecen trastorno psicomotor es mayor el riesgo de presentar estos déficits, lo cual se convierte en factor de agresión añadido a su estado general que puede tener graves consecuencias. Por su importancia e interés clínico en beneficio de los pacientes con trastorno psicomotor, se presenta este planteamiento de Evaluación Nutricional, que si bien tiene como base los valores estándar para individuos sanos, de edad y sexo similares, aporta algunas especificaciones útiles para la valoración de quienes presentan trastorno psicomotor. Esta evaluación será provechosa, siempre que se realice de modo sistemático y con una periodicidad determinada (AU)

Giant cell arteritis localized to the colon associated with Crohn's disease.

AIMS: Intestinal vasculitis is uncommon and usually accompanies systemic vasculitis. Although intestinal vascular changes including vasculitis have been studied intensively, and are found regularly in Crohn's disease, giant cell arteritis is distinctly unusual. We describe a case of giant cell arteritis localized to the colon of an 18-year-old girl suffering from Crohn's disease. METHODS AND RESULTS: After three years of medical treatment, the patient underwent a proctocolectomy. The medium-sized arteries of the mesocolon demonstrated striking changes characterized by intimal fibrous thickening and an inflammatory infiltrate with giant cells, most predominant at the intima-media junction. Epithelioid cells and sarcoid-like granulomas were not observed. The internal elastic lamina was fragmented. Neither clinical symptoms nor laboratory findings showed evidence of systemic vasculitis. Neither the chest CT scan nor the echo-Doppler of the temporal arteries, supra-aortic and abdominal vessels revealed any abnormality. CONCLUSIONS: This case illustrates an extremely rare feature in the spectrum of vascular lesions in Crohn's disease which have to be differentiated from temporal and Takayasu's arteritis.

Flow cytometric assessment of amphotericin B susceptibility in Leishmania infantum isolates from patients with visceral leishmaniasis.

Amphotericin B susceptibility was measured by a flow cytometric membrane potential assay in Leishmania infantum promastigotes isolated from 11 immunocompetent children treated with liposomal amphotericin B and 19 HIV-infected young adults treated with intralipid amphotericin B. Susceptibility levels were measured by the 90% inhibitory concentrations (IC90) representing the concentrations of drug that induced a 90% decrease in membrane potential compared with the control culture. In immunocompetent children, treatment was fully effective whatever the susceptibility of isolates to amphotericin B. In immunocompromised adults, on the contrary, unresponsiveness and relapses could be observed in all cases and IC90 increased in the course of successive treatments: a decrease of amphotericin B susceptibility in both promastigote and amastigote forms could be observed in a patient who had six relapses. These results suggest that the success of amphotericin B treatment depends greatly on patient immunity status, and indicate that successive relapses could enhance emergence of amphotericin B resistant isolates. The results demonstrate that the flow cytometric membrane potential assay can be used as an easy and reliable tool for studying the evolution of interactions between amphotericin B and the parasite membrane during long-term treatments.

Early progenitor cells from human mobilized peripheral blood express low levels of the flt3 receptor, but exhibit various biological responses to flt3-L.

The biological effects of flt3-L, and the expression of its tyrosine kinase receptor (flt3, CD135) were investigated on the immature subsets of human circulating peripheral blood progenitors obtained from cancer patients or normal volunteer donors, after mobilization with rhG-CSF or chemotherapy. flt3 was expressed at low levels, and its expression increased concomitantly with expression of CD38 within the CD34+ cell population. Despite this low-level expression, flt3-L exerted synergistic effects with a combination of c-kit ligand, IL-3, IL-6, GM-CSF and G-CSF, mainly to induce proliferation of CD34+/CD38- cells. In addition, flt3-L increased the detection of HPP-CFC, both immediately after cell selection, and after 7 and 14 d of cultures. We conclude that flt3-L is active on circulating early mobilized haemopoietic progenitors, despite the low- level expression of its receptor.

Chromosome region 8p11-p21: refined mapping and molecular alterations in breast cancer.

Several genes, most of them unknown, of the short arm of chromosome 8 are involved in malignant diseases. Numerous studies have implicated a portion of the 8p11-p21 region as the location of one or more tumor suppressor genes involved in a variety of human cancers, including breast cancer. We and others have reported linkage analyses suggesting the presence of a putative breast cancer susceptibility gene. Furthermore, several oncogenes of the 8p11-p12 region are involved in reciprocal translocations in myeloproliferative and myelodysplastic disorders and in amplification in breast cancer. To facilitate the analysis of the 8p11-p21 region and the cloning of candidate oncogenes and tumor suppressor genes, a high-resolution physical and transcriptional map was established with 39 yeast artificial chromosomes and 94 markers, including so-called sequence-tagged sites and expressed sequence-tagged sites derived from either known genes or expressed sequence tags corresponding to unidentified transcripts. In addition, four novel transcripts were identified and localized precisely within the map. This transcription map provides a detailed description of gene order for the 8p11-p21 region and will be helpful in the identification of candidate genes for diseases. From this basis, we refined the mapping of two types of molecular alterations that occur at 8p11-p21 in sporadic breast cancers, i.e., amplification and deletion.

A neutralizing anti-TGF-beta1 antibody promotes proliferation of CD34+Thy-1+ peripheral blood progenitors and increases the number of transduced progenitors.

The subset of blood cells that expresses both CD34 and Thy1 (CD90) cell surface molecules is enriched in hematopoietic stem cell activity and can be obtained from the peripheral blood of cancer patients after mobilization by chemotherapy and granulocyte colony-stimulating factor (G-CSF). Because transforming growth factor-beta1 (TGF-beta1) is a potent inhibitor of hematopoietic progenitor proliferation and differentiation, in this study we analyzed the impact of neutralizing TGF-beta1 activity during culture and retroviral transduction of CD34+Thy1+ cells. When purified CD34+Thy1+ cells were cultured in the presence of a neutralizing antibody against TGF-beta1, the percentage of cycling cells, proliferation, and absolute number of clonogenic progenitors were increased in comparison to the cultures performed without the addition of antibody. Antibody-mediated neutralization of TGF-beta1 during retroviral transduction performed by coculture of CD34+Thy1+ cells with a MFG-S-nlsLacZ retroviral vector-producing cell line did not affect the percentage of transduced progenitors as assessed by direct X-Gal staining of colonies in clonogenic assays. However, due to the better expansion of CD34+Thy1+ cells in the presence of anti-TGF-beta1, the absolute number of transduced progenitors recovered at the end of the culture was increased.

The ECG in pulmonary embolism. Predictive value of negative T waves in precordial leads--80 case reports.

BACKGROUND AND STUDY OBJECTIVE: The value of the ECG for the diagnosis of pulmonary embolism (PE) is debatable. Once the diagnosis of PE has been established, however, the ECG could allow the massive forms to be distinguished. The purpose of our study was to analyze the ECG signs in patients hospitalized for PE in a cardiology unit. DESIGN: Taking a series of 80 consecutive patients hospitalized for PE, we analyzed the ECGs on admission and then during hospitalization. We sought to evaluate changes in ECG signs compared with angiographic and hemodynamic changes in PE. RESULTS: T-wave inversion in the precordial leads is the most common abnormality (68%), and represents the ECG sign best correlated to the severity of the PE. Among those patients with anterior T-wave inversion, 90% had a Miller index over 50% (mean, 60 +/- 8%). Eighty-one percent had a mean pulmonary arterial pressure (PAP) over 30 mm Hg (mean, 37 +/- 8%). This subepicardial ischemic pattern is an even stronger marker of severity when it appears as early as the first day (p < 0.01). Its reversibility is correlated to the changes in PE. After thrombolysis in particular, normalization of repolarization systematically indicates mean Miller and PAP indexes of < 20% and < 20 mm Hg, respectively. CONCLUSIONS: The anterior subepicardial ischemic pattern is the most frequent ECG sign of massive PE. This parameter is easy to obtain and reflects the severity of PE. Its reversibility before the sixth day points to a good outcome or high level of therapeutic efficacy.

Efficiency of retroviral transduction into hematopoietic cells by cocultivation procedure does not correlate with viral titer.

Relative transduction efficiency with retroviral vector-producing clones was assayed by cocultivating TF-1, a human CD34+ hematopoietic cell line and YR-2, a sheep B-lymphoid cell line, with LacZ containing vector-producing cells, and then by scoring the percentage of X-Gal+ cells. At the same time, viral titer was estimated by titration assay with murine fibroblasts. Results clearly demonstrated a lack of correlation between viral titer and efficiency of transduction into hematopoietic cells, which depends neither on the type of packaging cell line, PG-13 and GP-envAM12 in this study, nor on the type of LacZ containing retroviral vector. These results strongly favor consideration of interactions between producers and target cells of the study for the screening of producing cell lines.

Novel sequence tagged sites from the human 8p12-p21 chromosomal region involved in breast cancer.

Seventeen sequence-tagged sites (STSs) from the human 8p12-p21 chromosomal region were identified in a subset of nonchimeric yeast artificial chromosomes (YACs). YAC ends were cloned, sequenced and a PCR primer pair was designed for each of the STSs thus generated. STSs were mapped by means of PCR on somatic cell hybrids specific for chromosome 8. These STSs will contribute to the construction of physical, high-resolution genetic and transcriptional maps necessary to clone potential tumor suppressor genes of the 8p12-p21 region involved in breast cancer.