14-3-3ε protein-immobilized PCL-HA electrospun scaffolds with enhanced osteogenicity.

Adipose-derived mesenchymal stem cells (ASCs) accelerate the osteointegration of bone grafts and improve the efficiency in the formation of uniform bone tissue, providing a practical and clinically attractive approach in bone tissue regeneration. In this work, the effect of nanofibrous biomimetic matrices composed of poly(ε-caprolactone) (PCL), nanometric hydroxyapatite (nHA) particles and 14-3-3 protein isoform epsilon on the initial stages of human ASCs (hASCs) osteogenic differentiation was investigated. The cells were characterized by flow cytometry and induction to differentiation to adipogenic and osteogenic lineages. The isolated hASCs were induced to differentiate to osteoblasts over all scaffolds, and adhesion and viability of the hASCs were found to be similar. However, the activity of alkaline phosphatase (ALP) as early osteogenic marker in the PCL-nHA/protein scaffold was four times higher than in PCL-nHA and more than five times than the measured in neat PCL.

Human papillomavirus infection in cyclosporin-induced gingival overgrowth in renal allograft recipients.

BACKGROUND: Host immunity plays an important role in the development of human papillomavirus (HPV)-associated disease. The HPV infection in oral cyclosporin-induced gingival overgrowth in renal transplant recipients has not been investigated previously. The aim of this study was to establish the HPV infection of cyclosporin-induced gingival hyperplasia in renal transplant recipients through morphological changes and use of the in situ hybridization technique. METHODS: We examined 13 renal transplant recipient biopsies with gingival overgrowth lesions and 4 healthy mucosa samples of these patients. The histopathological diagnoses were established on the basis of widely accepted criteria, and the pathologist was not aware of the HPV result. An in situ molecular hybridization was carried out under low stringent conditions to detect HPV species with mixed biotin-labeled probes of HPV 6 and HPV 11, and under high stringent conditions with HPV 6, HPV 11, HPV 16, and HPV 18 probes for HPV typing. RESULTS: The HPV prevalence among the 13 samples studied was 92.31% (12/13), of which 4 tested positive for HPV 6-11 and 1 for HPV 16. The 4 biopsies of normal mucosa from gingival overgrowth patients were also reactive for HPV DNA. In 11/12 (91.7%) HPV-positive cases, koilocytotic atypia was found. CONCLUSIONS: The suppression of T-cell function by cyclosporin therapy can result in an increase of HPV infection, adding to the proliferative activity of cyclosporin in the oral mucosa.

Conversion from azathioprine [correction of azathioprina] to mycophenolate mofetil in pediatric renal transplant recipients with chronic rejection.

BACKGROUND: Chronic rejection is the leading cause of graft failure. Both nonimmunological and immunological mechanisms contribute to this pathology. METHODS: We studied changes in kidney function, mixed lymphocyte culture, cell-mediated lympholysis, serum HLA class I antigens, cytotoxic antibodies, and lymphocyte population before and after 6 months of follow-up in 22 pediatric renal transplanted patients. The immunosuppressive protocol used was: cyclosporine, azathioprine, and corticosteroids. Eight patients demonstrated chronic graft rejection (by biopsy), group I; and eight patients had no clinical evidence of chronic and/or acute rejection, group II. Substitution of mycophenolate mofetil (MMF) (600 mg/m2 bid for azathioprine was done in patients of groups I and II. Another six patients with chronic rejection, did not receive MMF, group III. RESULTS: Creatinine clearance increased in group I (44+/-5 vs. 51.1+/- ml/min/1.73 m2, P<0.03) but it decreased in group III (30+/-3 vs. 25+/-2, P<0.01). Urinary protein excretion decreased only in group I (0.3+/-0.03 to 0.06+/-0.03 g/24 hr, P<0.03). During MMF therapy antidonor mixed lymphocyte culture decreased 62 and 70% (P<0.05) in group I and II. Cell-mediated lympholysis against lymphocyte of the donor decreased 65% (P<0.05) in group I. Cell-mediated lympholysis toward control cells decreased 54% (P<0.01) in group II. Serum HLA class I antigens, only decreased from 0.7+/-0.1 to 0.5+/-0.1 microl/ml, P<0.05, in group I. CD19+ decreased from 7.9+/-1.1 to 5.6+/-0.8%, P<0.05, and 7.8+/-1.2 to 5.5+/-0.9%, P<0.05, in groups I and II, respectively. CD16+ increased from 5.7+/-1.1 to 8.6+/-1.3 (P<0.05) only in group I. CONCLUSIONS: Our data suggest that substituting MMF for azathioprine therapy leads to an improvement in the immunosuppression and renal function in children with on-going chronic rejection.

Quantification of fecal neutrophils by MPO determination (myeloperoxidase) in patients with invasive diarrhea. Cuantificación de neutrofilos fecales mediante la determinación de MPO (Mieloperoxidasa) en pacientes con diarrea invasiva

Myeloperoxidase (MPO), a specific polymorphonuclear leukocyte enzyme, has been used previously to quantify the number of neutrophils in tissue. MPO activity was found to be linearly related to the number of neutrophil cells. In an attempt to use this method in leukocytes measuring in stool, fecal MPO was solubilized with hexadecyltrimethylammonium bromide and the MPO activity was measured by a dianisidine-H2O2 assay. Stools from 10 normal subjects and 39 patients with diarrhea produced by enteropathogenic bacteria were examined for leukocytes by MPO activity as well as microscopically using methylene blue stain, MPO activity was positive in 36 patients (92

) and leukocytes were present by microscopic observation in 30 (77

). Fecal leukocytes were not found in healthy controls and the MPO activity was undectable. Stool MPO activity had a range of from 1.6 to 2,830.0 x 10(3) UMPO per gram of feces (median 460.0). The number of neutrophils obtained through MPO activity had a range of 6.0 to 13,216.0/ mm3 (median 1,261.0). Fecal MPO activity is a simple biochemical assay for the detection and quantification of fecal leukocytes.

Cuantificación de neutrófilos fecales mediante la determinacián de MPO (Mieloperoxidasa) en pacientes con diarrea invasiva / Quantify of fecal neutrophils by MPO (myeloperoxidase) determination in patients with invasive diarrhea

La mieloperoxidasa (MPO) es una enzima especifíca de los polimorfonucleares neutrófilos, la cual ha sido previamente usada para cuantificar elmnúmero de neutrófilos en tejidos, desde que su actividad se correlaciona linealmente con el número de neutrófilos. Con el objetivo de demostrar la presencia y realizar la cuantificación de leucocitos en materia fecal la MPO se disolvió en Bromuro de hexadeciltrimetilamonio y la actividad fue medida usando un ensayo de H2O2O-dianosid-ina. Se midió la actividad de MPO en 39 pacientes con diarrea producida por bacterias enteropatógenas y en 10 sujetos control. La presencia de leucocitos fue también determinada mediante la observación microscópica usando azul de metileno. La actividad de MPO fue positiva en 36 (92 por ciento) de los pacientes y la observació microscópica resultó positiva en 30 (77 por ciento). En los sujetos control la actividad de MPO fue indetectable y no se encontraron leucocitos en material fecal. En los pacientes la actividad de MPO en materia fecal tuvo un recuento de 1.6 a 2830 x 10(3) UMPO por gramo de heces (mediana: 46.0). El número de neutrófilos obtenido a través de la actividad de MPO tuvo un recuento de 6 a 13216.0 x mm(3) (mediana: 1261.0), la actividad fecal de MPO es una determinación bioquímica simple para la detección y cuantificación de leucocitos en materia fecal.

Identification of mRNA-binding proteins during development: characterization of Bufo arenarum cellular nucleic acid binding protein.

Ultraviolet irradiation was used to covalently cross-link poly(A)+RNA and associated proteins in eggs and embryos of the toad Bufo arenarum. Four major proteins with apparent sizes of 60, 57, 45 and 30-24 kDa were identified. It was observed that the same mRNA-binding proteins were isolated from eggs to gastrula and neural stages of development. The 30 kDa polypeptide, p30, appeared as the main ultraviolet (UV) cross-linked protein in the developmental stages analyzed. By means of polyclonal antibodies, it was determined that this polypeptide has a cytoplasmic localization and it was detected in liver, eggs and embryos. The presence of p30 was also analyzed by western blot during oogenesis and development. The 30 kDa polypeptide was present in all stages analyzed but it could not be detected in stages I-II of oogenesis. At the neural stage, the relative amount of p30 began to decrease, reaching its lowest levels after stages 26-30 (tail-bud in Bufo arenarum). On the basis of purification, immunoprecipitation and western blot assays the 30 kDa protein was identified as the Bufo arenarum cellular nucleic acid binding protein.

[Human papillomavirus detection in oral cancer lesions in the city of Córdoba]. / Detección del virus papiloma humano en lesiones cancerosas orales en la ciudad de Córdoba.

Oral cancer is a process that involves different etiological factors and mechanisms in the light of current view of viral cocarcinogenesis. Evidence from histology and DNA hybridization studies suggests that HPV is engaged in oral carcinogenesis. The Pathology Laboratory of the Dentistry School, National University of Córdoba, admits approximately 20% of all patients with cancerous lesions in this city. In the January 1992-December 1997 lapse, we examined 1950 biopsies with oral lesions, 4.77% (93/1950) of which were malignant neoplasms, 79.57% (74/93) were oral carcinomas. Thirty-three oral carcinomas (44.6%; 33/74) were selected at random and included in this study, 33 cells smears of normal oral mucosa of controls individuals were included. They were analyzed by conventional light microscopy and an in situ hybridization technique for the detection of HPV. Data were analyzed with chi square test. The prevalence of HPV among the 33 cancer samples studied was 27.27%, 9/33 tested positive for HPV in low stringent conditions. Only one was positive in high stringent condition for HPV16, a verrugous carcinoma. No HPV-DNA was detected in cells smears of controls. Among the HPV positive, 3/9 (33.33%) were squamous carcinomas and 5/9 (55.56%) were verrugous carcinomas. Only one was a melanoma. Verrugous carcinoma was the carcinoma most associated with the HPV infection (x2 = 20.5; 95% level of confidence). This would indicate a major role of HPV in the pathogenesis of verrucous carcinomas. The viral prevalence found in cancerous lesions reinforces the concept of heterogenic natures of oral cancer. HPV is a circumstance that increase the probability of malignancy, and when reducing, diminish the frequency of cancer.

Las formas elementales de la enfermedad: la sistematización de Laplantine / Elemental forms of the illness: the Laplantine systematization

En este artículo, se analiza el importante aporte de Francois Laplantine en el estudio de la antropología de la enfermedad, vía de acceso a la antropología de la salud en un sentido amplio. La importancia de este trabajo es que va más allá del análisis de los sistemas médicos tradicionales, sistematizando los discursos actuales válidos para la cultura occidental sobre el proceso salud-enfermedad, revelando la influencia de los factores culturales sobre el comportamiento y creencias de los enfermos, fundamental para mejorar la eficacia de los programas médicos en nuestros países latinoamericanos, cuya cultura básica es sincrética y oral. Al proponerse el concepto "Antropología de la enfermedad", se permite haciendo de la enfermedad un objeto de análisis ampliar el horizonte de custionamiento antropológico sobre la naturaleza del hombre y la cultura, antecedente básico para el desarrollo de una reflexión bioética en nuestro continente que repose en el "hombre total", no desinserto de su cultura y de su historia

Crecimiento de niños en zonas rurales / Growth of chidren in rural zones

El propósito de este estudio fue analizar la evolución del crecimiento en estatura de escolares de talla baja. Mediante rastreo, en escolares de 6 a 8 años de zonas rurales de alta vulnerabilidad social de las regiones Metropolitana y Sexta de Chile, se seleccionaron 127 niños con déficit de talla para la edad entre menos 1 y menos 3 puntajes Z de la mediana del patrón NCHS/OMS (casos) y 130 controles cuya estatura estaba entre más o menos medio puntaje Z. Se analizó la información de las fichas de los consultorios donde eran controlados. Se observó un incremento menor de estatura en los casos antes y después de la edad de 24 meses (p <0,001). La mayor des-aceleración de la talla ocurrió en el primer año de vida. Los casos mostraban valores inferiores a puntajes Z (-0,65 vs -0,13) desde el nacimiento, llegando a -1,72 alos 18 meses y a -1,83 a los 72 meses. Los controles se mantuvieron alrededor de la mediana durante este mismo período. La desnutrición y talla baja al nacer se asociaron significativamente con talla baja en el escolar (ventaja u OR= 4,94 y 3,36 respectivamente). Estos resultados sugieren la conveniencia de reforzar las acciones de prevención y control prenatales y posnatales si se desea disminuir la frecuencia de talla baja en los escolares chilenos

Prevention of asthma with ketotifen in preasthmatic children: a three-year follow-up study.

Growing morbidity and mortality rates call for research towards more effective methods of preventing asthma. During the last decade several groups have reported the results of natural history and asthma prevention studies. However, the attempt to prevent development of asthma in genetically predisposed children, has not resulted in a generally accepted management scheme. The aim of this study was to evaluate the effectiveness of ketotifen in preventing the onset of asthma in infants considered to be at high risk of developing the disease, but who had no history of respiratory obstruction. These children have been described as preasthmatic. In this double-blind, placebo-controlled, parallel study, 100 infants with a family history of major allergy and elevated serum IgE levels, but with no history of bronchial obstruction, were treated with either ketotifen (n = 50) or placebo (n = 50) over a 3-year period. There were no statistically significant differences between the two groups with regard to age, sex, degree of hereditary allergy, levels of serum IgE upon joining the study, and family smoking habits. At the end of 3 years, only four of the 45 infants who had received ketotifen had developed asthma (9%). Of the 40 children given placebo, 14 had developed asthma (35%) (P = 0.003). These results suggest that ketotifen is effective in preventing the onset of asthma in preasthmatic children.

Influencia de la ingesta de butirato de sodio en la carcinogénesis colónica murina / Influence of sodium butyrate intake on murine colonic carcinogenesis

El papel de los ácidos grasos de cadena corta (AGCC) en la carcinogénesis colónica murina (CCM) no fue aclarado. Evaluamos el efecto de la hemicolectomía derecha (HCD) (colon derecho, fuente de AGCC) y de la ingesta en agua de bebida de butirato de sodio (Buti.Na) al 2 por ciento a pH 7 o de cloruro de sodio (CINa) 4g/l, en la CCM. Formamos 7 grupos de 12 ratas Wistar macho de 150 g: HCD, Buti. Na, CINa, control (C). La mitad recibió dimetilhidrazina (DMH) 20 mg/Kg subucutánea semanal durante 12 semanas. La necropsia fue realizada a los 6 meses. Determinamos el contenido de AGCC en materia fecal por cromatografía gaseosa. El 70 por ciento de las ratas con DMH desarrolló tumor. El n§ de animales con tumor, por grupo fue: HCD 4/6, Buti.Na 4/6, CINa 3/5, C 6/6. El n§ de tumores promedio por animal, por grupo fue: HCD 1,17 ñ 0,48, Buti.Na 3/5, C 6/6. El n§ de tumores promedio por animal, por grupo fue: HCD 1,17 ñ 0,48, Buti.Na 1,50 ñ 0,76, ClNa 1,20 ñ 0,49, C 1,50 ñ 0,22. El grupo Buti.Na (DMH) presentó una concentración significativamente menor de butirato (p < 0,05) en relación a los demás grupos. En conclusión, el suplemento en agua de bebida de Buti.Na, CINa y la HCD redujeron en forma no significativa la CCM, con este número de animales

[Postmortem diagnosis of cardiac amyloidosis in the aged. Anatomoclinical correlation]. / Diagnostic post-mortem de l'amyloïdose cardiaque du vieillard. Corrélation anatomoclinique.

The absence of specific clinical signs makes the diagnosis of cardiac amyloidosis difficult. Moreover, it is established that this condition, the prevalence of which increases with age, aggravates the prognosis of cardiac failure. The present study was undertaken to identify the clinical or paraclinical signs enabling more accurate diagnosis of this disease. Analysis of 2589 autopsy reports from the University Institutes of Geriatrics of Geneva between January 1972 and January 1990 recensed 58 cases of microscopic cardiac amyloidosis, but this diagnosis was not made in any of these patients before death. Of the potential indicators, the good specificity but poor sensitivity of atrial fibrillation and low voltage electrocardiogram was confirmed. On the other hand, the author's research found the association of radiological cardiomegaly and a raised erythrocyte sedimentation rate in nearly 70% of cases of cardiac amyloidosis with a false positive rate of only 10% in a control group.

[Influence of sodium butyrate intake on murine colonic carcinogenesis]. / Influencia de la ingesta de butirato de sodio en la carcinogénesis colónica murina.

The role of short chain fatty acids (SCFA) in murine colonic carcinogenesis (MCC) has not yet been clarified. In rats, Freeman et al have reported an increased number of colonic tumors induced with dimethylhydrazine (DMH) and sodium butyrate in drinking water. On the other hand, Deschner et al showed that tributyrin intake did not increase MCC induced with azoxymethane. Both of them have reported high levels of fecal butyric acid with sodium butyrate and tributyrin intake. Although salt intake has been positively associated with colorectal cancer some authors do not support this association. We have evaluated the influence of right hemicolectomy (RH) (right colon as main source of SCFA) and the intake of 2%-pH 7 sodium butyrate (S.BUT) and 4 g/l sodium chloride (S.CHL) in drinking water, in MCC. Forty eight male Wistar rats weighing 150 g were divided into 4 groups: RH, S.BUT, S.CHL, control (C). Half of the animals received weekly DMH 20 mg/kg subcutaneously for 12 weeks. Necropsy was performed after 6 months. We have determined fecal SCFA content by gas chromatography. Neoplasm was present in 70% of rats treated with DMH. The number of animals with tumors was: RH 4/6, S.BUT 4/6, S.CHL 3/5, C 6/6. Tumor frequency was: RH 1.17 +/- 0.48, S.BUT 1.50 +/- 0.76, S.CHL 1.20 +/- 0.49, C 1.50 +/- 0.22. S.BUT group, treated with DMH, presented a lower butyric acid concentration (p < 0.05) in comparison with other groups. We have no explanation for this finding; gastric absorption of sodium butyrate may be an important factor.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of right hemicolectomy on nitrogen balance in the rat.

Rats with right hemicolectomy were divided into two groups. One group was free-fed (RHFF), the other group pair-fed (RHPF) and both were compared with a group of control animals. The nitrogen balance (NB) was studied for 5 days at 2 and 14 weeks after surgery. The two groups of rats with right hemicolectomy excreted significantly more fecal nitrogen than the control group (p less than 0.01) during the two periods of study. At 2 weeks the RHFF group increased N intake so that their NB was similar to that of the control group. In the same period the nitrogen lost in urine and the NB were reduced in the RHPF rats (p less than 0.01). At 14 weeks the results were similar to those for 2 weeks although the increase in food intake for RHFF and the decrease in NB for RHPF were not statistically significant. The rats with right hemicolectomy showed a decreased gain in body weight. This decrease was more pronounced in the RHPF group. The nitrogen content of the carcass and liver was significantly reduced in the RHPF group 4 months after surgery. It is concluded that the rats with right hemicolectomy were characterized by an increment in the nitrogen lost in feces, and when they were only allowed the same intake of food as the control group, the animals showed marked abnormalities in growth and in the nitrogen content of the organism.