Prescription patterns of somatostatin analogs in patients with acromegaly and neuroendocrine tumors.

PURPOSE: Acromegaly and neuroendocrine tumors are rare diseases that, under certain conditions, can be treated with somatostatin analogs. The aim was to determine the prescription patterns of somatostatin analogs in a group of patients with acromegaly and neuroendocrine tumors affiliated with the Colombian Health System. METHODS: A retrospective study. A cohort of patients from a drug dispensing database that collected all prescriptions of long-acting somatostatin analogs (octreotide, lanreotide, pasireotide). Sociodemographic variables, clinical variables (diagnosis and comorbidities) and pharmacological therapy variables (dose, changes, persistence of use, comedications) were considered. RESULTS: A total of 213 patients were identified, including 139 (65.3%) with acromegaly and 74 (34.7%) with neuroendocrine tumors. There was a predominance of women (58.7%) and a mean age of 59.7 ± 14.5 years. The most commonly used medications were lanreotide autogel (n = 107; 50.2%), octreotide LAR (n = 102; 47.9%) and pasireotide LAR (n = 4; 1.9%). During follow-up, 11.3% of patients experienced modifications of therapy, with a mean duration from the beginning of treatment to the change in medication of 25 ± 15.9 months. A total of 48.9% of the patients with acromegaly and 87.1% of individuals with neuroendocrine tumors received maximum approved doses of the drug. CONCLUSION: Patients with acromegaly and neuroendocrine tumors in Colombia are mainly women and are most frequently treated with lanreotide autogel for acromegaly and with octreotide LAR for neuroendocrine tumors. In addition, a high proportion are managed with maximum doses of long-acting somatostatin analogs.

Properties of antibacterial nano textile for use in hospital environments / Propiedades del nanotextil antibacteriano para su uso en entornosm hospitalarios / Propriedades do nano têxtil antibacteriano para seu uso em meios hospitalares

Abstract In hospital environments, there are several problems related to Healthcare-Associated Infections (HAIs), contaminated hospital textiles, can contribute to the spread and transmission of (HAIs), due to retention of viruses and bacteria. The antibacterial metallic nanoparticles immersed in hospital textiles can allow reduction of microorganisms. This paper presents a technological surveillance of the principal properties of antibacterial nanotextiles to be used in hospital environments, based on international standards. Initially, the search equation was determined for "antibacterial" AND "nanoparticle." Subsequently, the main properties were selected, by means of a multiple authors' review. Afterwards, the properties were related to international standards. Finally, we present the results found associated to the materials used to develop nonwoven textiles, and their properties for hospital environments, the sizes of samples and also the equipment required for characterization.

Resumen En los ambientes hospitalarios, existen varios problemas relacionados con las infecciones asociadas a la atención de la salud (HAI, por sus siglas en inglés), los tejidos hospitalarios contaminados, pueden contribuir a la propagación y transmisión de los HAIs, debido a la retención de virus y bacterias. Las nanopartículas metálicas antibacterianas sumergidas en tejidos hospitalarios permiten reducir los microorganismos. Este documento presenta una vigilancia tecnológica de las principales propiedades del nanotextil antibacteriano para uso en ambientes hospitalarios, basados en estándares internacionales. Inicialmente, la ecuación de búsqueda se determinó "antibacteriano" Y "nanopartícula". Posteriormente, se seleccionaron las principales propiedades, mediante la revisión de diferentes autores. Luego, las propiedades se relacionaron con los estándares internacionales. Finalmente, se presentan los resultados encontrados asociados a los materiales utilizados para el desarrollo de materiales no tejidos y sus propiedades para ambientes hospitalarios, tamaños de muestras y también el equipo necesario para la caracterización.

Resumo Nos ambientes hospitaleiros, existem vários problemas relacionados com as infecções associadas à atenção da saúde (HAI), os tecidos hospitalários contaminados, podem contribuir à propagação e transmissão dos HAIs, devido à retenção de vírus e bactérias. As nano partículas metálicas antibacterianas submergidas em tecidos hospitalários permitem reduzir os microorganismos. Este documento apresenta uma vigilância tecnológica das principais propriedades do nano têxtil antibacteriano para uso em ambientes hospitalários, baseados em padrões internacionais. Inicialmente, a equação de busca determinou-se "antibacteriano" e "nano partícula". Posteriormente, selecionaram-se as principais propriedades, mediante a revisão de diferentes autores. Posteriormente, as propriedades relacionaram-se com os padrões internacionais. Finalmente, apresentam-se os resultados encontrados associados aos materiais utilizados para o desenvolvimento de tecidos não tecidos e suas propriedades para ambientes hospitaleiros, tamanhos de amostras e também a equipe necessária para a caracterização.

Diversidad de micorrizas arbusculares en pasto colosuana (Bothriochloa pertusa (L) A. Camus de fincas ganaderas del municipio de Corozal-Sucre / Diversity of arbuscular mycorrhizae in grass colosuana (Bothriochloa pertusa (L) A. Camus of livestock farms of the municipality of Corozal- Sucre

Objetivo. Caracterizar comunidades de hongos formadores de micorrizas arbusculares (HMA) en rizosfera del pasto Colosuana (Bothriochloa pertusa (L). A. Camus) en fincas ganaderas del municipio de Corozal, departamento de Sucre. Materiales y métodos. Se muestrearon 43 fincas ganaderas distribuidas en 4 zonas agroecológicas del municipio. Se realizó aislamiento de esporas del suelo, determinación del porcentaje de colonización en raíces, separación e identificación de morfotipos de HMA. Resultados. El ANOVA multifactorial mostró que el número de esporas de HMA y el porcentaje de colonización en raíces de Colosuana (Bothriochloa pertusa), fueron mayores en las fincas localizadas en la zona agroecológica 4 y 2, respectivamente. 31 morfotipos de HMA fueron aislados, un 96.9% correspondieron a morfotipos con características similares a especie dentro del género Glomus y 3.1% a Gigaspora. Conclusiones. La alta diversidad de morfotipos de HMA asociados a Colosuana se convierte en un recurso biológico alternativo, cuyo manejo, conservación y efectos sobre la productividad, genera beneficios ambientales al mejorar las condiciones físico-químicas y biológicas del suelo. Se propone su inclusión en el inventario de la diversidad de microorganismos en estos agroecosistemas.

Objective. Characterize communities of fungi that produce arbuscular mycorrhizal (AMF) in the rhizosphere of the Colosuana grass (Bothriochloa pertusa (L). A. Camus) in cattle farms located in the municipality of Corozal, Sucre. Materials and methods. 43 cattle farms were sampled in 4 agro-ecological zones of the municipality. Spore isolation from the ground was performed, determining the percentage of root colonization, separation and identification of morphotypes of AMF. Results. The multifactor ANOVA showed that the number of AMF spores and the percentage of root colonization of Colosuana (Bothriochloa pertusa) was higher on farms located in the agro-ecological zone 4 and 2, respectively. 31 AMF morphotypes were isolated; a 96.9% corresponded to morphotypes with characteristics similar to species within the genus Glomus and 3.1% to Gigaspora. Conclusions. The high diversity of morphotypes of AMF associated Colosuana become an alternative biological resource, and its management, conservation and effects on productivity, generate environmental benefits by improving the physic-chemical and biological conditions of the soil. Its inclusion in the inventory of the diversity of microorganisms in these agro ecosystems is proposed.

Evaluation of the ability of a feed additive to ameliorate the adverse effects of aflatoxins in turkey poults.

1. A study was conducted to evaluate the possible protective effect of a feed additive containing aluminosilicate and phytogenic substances against the adverse effects of aflatoxins in turkey poults. 2. Dietary treatments (6) were given to turkey poults from d 1 to d 42 of age. From d 1 to 21 the dietary treatments were as follows: 1, negative control, no aflatoxins or feed additive added; 2, feed additive control, 1 kg/t feed additive, no aflatoxins; 3, 250 ppb (microg/kg) aflatoxins, no feed additive; 4, 250 ppb aflatoxins + 1 kg/t feed additive; 5, 500 ppb aflatoxins, no feed additive; and 6, 500 ppb aflatoxins + 1 kg/t feed additive. From d 22 to 42, the dietary concentration of the feed additive was increased from 1 to 2 kg/t in all treatment groups receiving the feed additive (2, 4 and 6), while keeping constant the dietary concentrations of aflatoxins. 3. Aflatoxins at 250 ppb did not cause adverse effects on performance but affected certain toxicopathological parameters. At 500 ppb, adverse effects on performance and several toxicological parameters were observed. 4. Some of the adverse affects were partially or completely overcome by supplementation with the feed additive, including amelioration of the performance parameters, suppression of mortality and correction of the immunological alterations induced by the exposure to the aflatoxins.

Dorsal carpometarcarpal dislocation of ulnar fingers. A case report.

Multiple dorsal dislocations of the carpometacarpal joints are rare. Diagnosis requires a high index of suspicion, careful examination and good radiography. Treatment is controversial and is based on the presence or not of associated fractures. This paper presents a case of combined fifth and fourth finger carpometacarpal joint dorsal dislocation, successfully treated with closed reduction and percutaneous pinning. A discussion of this pattern of injury is also presented.

Infección por Helicobacter pylori en pacientes con patología gastrointestinal benigna / Infection by Helycobacter pylori in patients with benign gastrointestinal patology

El objetivo de este trabajo es establecer asociación entre infección por Helicobacter Pylori y presenciade gastropatías benignas en indiviuos sintomáticos de la ciudad de Maracaibo. Se estudiaron 80 pacientes mediante prueba rápida de ureasa, cultivo bacteriológico y serología (IgG e IgA). A 46 de ellos se les realizó también PCR. La frecuencia de infección en 53,8 por ciento, en contraste a una seroprevalencia del 71,3 por ciento. Veintitrés de los individuos (28,75 por ciento) resultaron negativos a las diferentes pruebas diagnósticas. La infección se detectó en todos los casos de ulceración duodenal (6/6); el 64 por ciento de las DNU (16/25); el 50 por ciento de las úlceras gástricas (6/12) y el 40,5 por ciento de las gastritis (15/37). En los individuos estudiados, la infección por H. pylori se asocia fuertemente al desarrollo de patologías digestivas benignas, especialmente a enfermedad ulcero-péptica duodenal. Se recomienda continuar las investigaciones, para esclarecer el papel etiológico del microorganismo en patologías como la DNU

Resistencia a antimicrobianos y presencia de plásmidos en cepas de Escherichia coli aisladas de aguas residuales crudas y tratadas por lagunas de estabilización con fines de reuso en agricultura / Resistance to antimicrobial agents and plasmids presence in Escherichia coli strains fron raw and treated waters of the stabilization pounds for reuse in agriculture

Se determinó resistencia a antibióticos y presencia de plásmidos en cepas de Eschericia coli recuperadas de aguas crudas y tratadas de un sistema de lagunas de estabilización. Se determinó la resistencia a los siguientes antibióticos: ácido nalidíxico (Na), ampicilina (A), amikacina (AK), cloranfenicol (C), carbenicilina (CB), trimetropin (Tr), tobramicina (Tb), tetraciclina (T), gentamicina (G) y Kanamicina (K). La extracción de plásmidos se realizó por el método de lisis alcalina. El 81,25 por ciento de las cepas de aguas crudas fueron resistentes a A, el 68,75 por ciento a T, el 46,87 por ciento a Cb, el 12,50 por ciento a Tr y el 3,12 por ciento a C y a Tb. En el 83,87 por ciento de las cepas de aguas tratadas, fueron resistentes A, el 16,12 por ciento a Tr y a T, el 12,9 por ciento a Cb y el 3,22 por ciento a AK, C y Tb. El 100 por ciento de las cepas fueron sensibles a Na, G y K. El orden de resistencia en las muestras crudas y tratadas fue: A>Tb>Tr>Cb>AK=C=Tb y A>T>Cb>Tr>C=Tb, respectivamente. El 16,12 por ciento y el 34,2 por ciento de las cepas de aguas residuales crudas y tratadas, respectivamente, presentaron resistencia a más de tres antimicrobianos. La presencia de plásmidos se detectó en el 87,10 por ciento de las cepas de aguas crudas y en el 90,62 por ciento de las cepas de aguas tratada. Las cepas de E. coli aisladas a la salida del sistema de tratamiento y presentaron plásmidos y resistencia a 7 de 10 antimicrobianos, lo que sugiere el intercambio genético entre las cepas y su potencial como patógenos, al combinar diferentes propiedades que contribuyan a su sobrevivencia

Cloning and characterization of a second acid phosphatase from Sinorhizobium meliloti strain 104A14.

Sinorhizobium meliloti has two nonspecific periplasmic acid phosphatases. The NapD enzyme has been previously described, and a second acid phosphatase, NapE, is described in this report. NapE was partially purified from an S. meliloti napD mutant and characterized with respect to molecular mass and substrate range. As predicted from SDS-PAGE analysis, the subunit molecular mass of NapE is approximately 35.8 kDa and gel filtration experiments estimated the native molecular mass to be approximately 70 kDa, indicating that the active enzyme is a homodimer. NapE demonstrated significant activity with p-nitrophenyl phosphate, phenyl phosphate, and alpha-naphthyl-phosphate. The pH optimum was between 4.5 and 5.0. The gene encoding NapE was also sequenced and the inferred amino acid sequence from the predicted ORF was found to be 60% identical and 75% similar to that encoded by napD. An S. meliloti napE mutant was constructed and assessed for symbiotic competence. This mutant did not differ from the wild-type parent strain in nodulation and symbiotic efficiency.

The role of airway stents in the management of pediatric tracheal, carinal, and bronchial disease.

OBJECTIVE: A variety of stents are available to aid in the management of complex tracheal, carinal and bronchial stenoses. We reviewed our multi-institutional experience with airway stents in children. METHODS: Thirty-three children (age, 13 days-18 years) from four institutions have had a total of 40 stents placed to aid in the management of complex airway stenoses. Three stent types were utilized: 29 silastic stents, five expandable metal stents and six customized carinal stents (four patients had two stents and one patient had four stents). Thirty children had tracheal stents, six children had bronchial stents, and two infants had carinal stents (three children had stenting of more than one area and two had stenting of all three locations). Twenty-eight patients (age, 5 months-18 years; mean, 8.06 years; SEM, 1.13 years) had stents placed after a variety of airway reconstructive procedures. Four underwent stenting in a non-operative setting and one as preoperative stabilization. RESULTS: Twenty-seven patients survived. One patient died early due to bleeding. Five patients died late: two due to bleeding, one from mediastinitis, and two patients with functional airways died late from unrelated problems. Complications are related to stent type and location. Carinal stents can migrate; several techniques are available to help manage this problem. Wire stents are essentially non-removable requiring periodic dilation. Silastic stents stimulate granulation tissue formation requiring periodic bronchoscopic removal. CONCLUSION: Tracheal stenting can aid in the management of pediatric airway problems. Complications are common, but can be managed with appropriate intervention.

The ++Sinorhizobium meliloti lon protease is involved in regulating exopolysaccharide synthesis and is required for nodulation of alfalfa.

While screening for Sinorhizobium meliloti Pho regulatory mutants, a transposon mutant was isolated that constitutively expressed higher levels of acid and alkaline phosphatase enzymes. This mutant was also found to form pseudonodules on alfalfa that were delayed in appearance relative to those formed by the wild-type strain, it contained few bacteroids, and it did not fix nitrogen. Sequence analysis of the transposon insertion site revealed the affected gene to have high homology to Lon proteases from a number of organisms. In minimal succinate medium, the mutant strain was found to grow more slowly, reach lower maximal optical density, and produce more extracellular polysaccharide (EPS) than the wild-type strain. The mutant fluoresced brightly on minimal succinate agar containing calcofluor (which binds to EPSI, a constitutively expressed succinoglycan), and gas chromotographic analysis of purified total EPS showed that the glucose-to-galactose ratio in the lon mutant total EPS was 5.0 +/- 0.2 (mean +/- standard error), whereas the glucose-to-galactose ratio in the wild-type strain was 7.1 +/- 0.5. These data suggested that in addition to EPSI, the lon mutant also constitutively synthesized EPSII, a galactoglucan which is the second major EPS known to be produced by S. meliloti, but typically is expressed only under conditions of phosphate limitation. (13)C nuclear magnetic resonance analysis showed no major differences between EPS purified from the mutant and wild-type strains. Normal growth, EPS production, and the symbiotic phenotype were restored in the mutant strain when the wild-type lon gene was present in trans. The results of this study suggest that the S. meliloti Lon protease is important for controlling turnover of a constitutively expressed protein(s) that, when unregulated, disrupts normal nodule formation and normal growth.

Characterization of two inducible phosphate transport systems in Rhizobium tropici.

Rhizobium tropici forms nitrogen-fixing nodules on the roots of the common bean (Phaseolus vulgaris). Like other legume-Rhizobium symbioses, the bean-R. tropici association is sensitive to the availability of phosphate (P(i)). To better understand phosphorus movement between the bacteroid and the host plant, P(i) transport was characterized in R. tropici. We observed two P(i) transport systems, a high-affinity system and a low-affinity system. To facilitate the study of these transport systems, a Tn5B22 transposon mutant lacking expression of the high-affinity transport system was isolated and used to characterize the low-affinity transport system in the absence of the high-affinity system. The K(m) and V(max) values for the low-affinity system were estimated to be 34 +/- 3 microM P(i) and 118 +/- 8 nmol of P(i) x min(-1) x mg (dry weight) of cells(-1), respectively, and the K(m) and V(max) values for the high-affinity system were 0.45 +/- 0.01 microM P(i) and 86 +/- 5 nmol of P(i) x min(-1) x mg (dry weight) of cells(-1), respectively. Both systems were inducible by P(i) starvation and were also shock sensitive, which indicated that there was a periplasmic binding-protein component. Neither transport system appeared to be sensitive to the proton motive force dissipator carbonyl cyanide m-chlorophenylhydrazone, but P(i) transport through both systems was eliminated by the ATPase inhibitor N,N'-dicyclohexylcarbodiimide; the P(i) transport rate was correlated with the intracellular ATP concentration. Also, P(i) movement through both systems appeared to be unidirectional, as no efflux or exchange was observed with either the wild-type strain or the mutant. These properties suggest that both P(i) transport systems are ABC type systems. Analysis of the transposon insertion site revealed that the interrupted gene exhibited a high level of homology with kdpE, which in several bacteria encodes a cytoplasmic response regulator that governs responses to low potassium contents and/or changes in medium osmolarity.

Prevalencia de enteroparasitosis en individuos que acuden a la unidad docente asistencial de medicina familiar "Luis Sergio Pérez" / Prevalence of intestinal parasites in individuals attending

Establecer la prevalencia de enteroparásitos en individuos que asistieron a la Unidad Docente Asitencial de Medicina Familiar "Luis Sergio Pérez". Comparar la recuperación de Enteroparásitos a través del método directo y el método concentrado. Se analizaron 100 muestras de heces a través del método directo y el método concentrado de Ritchie. Se practicó la coloración de Zielh-Neelsen modificada a las muestras diarreicas y a la Técnica de Kato-Katz a aquellas muestras donde se observaron huevos de helmintos para determinar la intensidad geohelmíntica. Se obtuvo una prevalencia de infecciones parasitarias del 64 por ciento. La mayor casuística se observó en el sexo femenino (73 por ciento) y en el grupo etario mayor de 15 años (67,2). El porcentaje de parásitos patógenos detectados en las heces de los individuos examinados fue: Blastocystis hominis (38.7 por ciento), Giardia lamblia (14.2 por ciento), Entamoeba histolytica (12,2 por ciento), Trichuris trichiura (6,1 por ciento) y Ascaris lumbricoides (1,0 por ciento). Se encontró un 68 por ciento de coincidencia entre los resultados obtenidos por el método directo y la técnica de concentración de Ritchie. El 25 por ciento resultó postivo por el método concentrado y el 7 por ciento fue positivo para el directo. El empleo del Chi cuadrado con un nivel de significancia de 0,05 demostró ser significativo para B.hominis (p=0,000) y G.lamblia (p=0,000). Se obtuvo una elevada prevalencia de enteroparásitos en relación a las condiciones satisfactorias de saneamiento que caracterizan a la población en estudio. Los parásitos predominantes fueron: B.hominis y G.lamblia. El método de concentración resultó más efectivo que el método directo

Quistes de giardia en aguas negras / Giardia cysts in raw sewage

El estudio de las aguas negras ha sido propuesto para conocer el estatus de infección de una comunidad y para indicar la presencia y variación estacional de los agentes infecciosos. Esta investigación fue llevada a cabo para evaluar un método para determinar la concentración de quistes de Giardia en aguas negras. En una primera etapa se realizó la prueba de eficiencia de recuperación de quistes siguiendo el método de floculación con carbonato de calcio descrito por Vesey y Cols., 1993, a partir de muestras estériles de aguas negras y agua destilada sembradas con una concentración conocida de quistes. En una etapa siguiente fueron analizadas muestras de aguas negras. Los quistes de Giardia fueron cuantificados con un hemocitómetro luego de la concentración de estos en solución de sacarosa (gravedad específica 1,15) o mediante centrifugación simple. La eficiencia de recuperación osciló entre 66 por ciento y 75 por ciento para las muestras de aguas negras y agua destilada respectivamente. Las recuperaciones mayores (87 por ciento) fueron obtenidas cuando se omitió el paso de clarificación. Las concentraciones de quistes de Giardia estuvieron entre 2 por diez elevado a la tres y 3.7 por diez elevado a la cuatro quistes/ml

Follicular atresia as an apoptotic process: atresia-associated increase in the ovarian expression of the putative apoptotic marker sulfated glycoprotein-2.

OBJECTIVE: To evaluate the possibility that morphologically confirmed/hypophysectomy-induced ovarian follicular atresia, a putative apoptotic process, is coupled to alterations in the steady-state levels of ovarian sulfated glycoprotein-2 (SGP-2) transcripts. METHODS: Hypophysectomy-induced follicular atresia in immature rats, morphologically confirmed at the light and electron microscopic levels, was correlated with alterations in the steady-state levels of ovarian SGP-2 transcripts as assessed by a solution hybridization/RNase protection assay. Cellular localization was accomplished by in situ hybridization technology. RESULTS: Hypophysectomy of the 24-day-old immature rat, an established precipitant of follicular atresia, led (3 days later) to a significant (P < .05) increase (up to 3.3-fold) in the relative abundance of densitometrically quantified ovarian SGP-2 transcripts compared with age-matched intact controls. Detailed time-course analysis after hypophysectomy revealed significantly (P < .05) increased ovarian SGP-2 mRNA expression as early as 2 days after hypophysectomy; no further increments were noted on days 4 or 8. Light microscopic analysis of comparable ovarian material 4 days after hypophysectomy revealed increased numbers of atretic follicles displaying large numbers of degenerating granulosa cells. Electron microscopic analysis of the degenerating cells of atretic follicles (from hypophysectomized rats) disclosed nuclear condensation and cytoplasmic shrinkage as well as apoptotic bodies at all levels of the granulosa cell layer. In situ hybridization established the granulosa cell of the intact untreated rat as the somatic cell concerned with SGP-2 gene expression. In turn, hypophysectomy led to an increase in SGP-2 expression at the level of the theca-interstitial cell, an effect prevented by the concurrent provision of pregnant mare serum gonadotropin (PMSG). The hypophysectomy-induced increase in ovarian SGP-2 transcripts was similarly reversed (54% inhibition by day 27) by the concomitant provision of FSH, an established antiatretic principle. The delayed administration (day 26) of a single dose of PMSG to rats hypophysectomized on day 24 eliminated the hypophysectomy-induced increase in ovarian SGP-2 transcripts as assessed on day 28. Qualitatively similar but quantitatively more pronounced increments in ovarian SGP-2 gene expression were obtained when atresia was induced by hypophysectomy of PMSG-primed immature rats. CONCLUSIONS: These observations establish the immature rat ovary as a site of SGP-2 gene expression and reveal hypophysectomy-induced follicular atresia to result in the up-regulation of ovarian (specifically, theca-interstitial) SGP-2 gene expression, an effect prevented by the concurrent provision of FSH or PMSG. To the extent that SGP-2 is an acceptable apoptotic marker, the present findings support the hypothesis that ovarian follicular atresia may be an apoptotic process.

Severe homozygous protein C deficiency: identification of a splice site missense mutation (184, Q-->H) in exon 7 of the protein C gene.

Single strand conformation polymorphism (SSCP) analysis of exon 7 of the protein C gene has identified a novel splice site missense mutation (184, Q-->H), in a newborn child with purpura fulminans and undetectable protein C levels. The mutations, seen in the homozygous state in the child and in the heterozygous state in her mother, was characterized and found to be a G to C nucleotide substitution at the -1 position of the donor splice site of intron 7 of the protein C gene, which changes histidine 184 for glutamine (184, Q-->H). According to analysis of the normal and mutated sequences, this mutation should also abolish the function of the donor splice site of intron 7 of the protein C gene. Since such a mutation is compatible with the absence of gene product in plasma and since DNA sequencing of all protein C gene exons in this patient did not reveal any other mutation, we postulate that mutation 184, Q-->H results in the absence of protein C gene product in plasma, which could be the cause of the severe phenotype observed in this patient.

Insulin-like growth factor I gene expression by primary cultures of ovarian cells: insulin and dexamethasone dependence.

A growing body of information now supports the existence of a complete intraovarian insulin-like growth Factor I (IGF-I) system replete with ligands, receptors, and binding protein(s). However, studies concerned with the regulation of ovarian IGF-I gene expression remain scarce. It was thus the objective of this communication to evaluate the expression of the IGF-I gene in the immature rat ovary under in vitro conditions. Whole ovarian dispersates or isolated granulosa cells were cultured for up to 96 h under serum-free conditions in the absence or presence of the indicated experimental agents. Extracted total RNA was subjected to a sensitive solution hybridization/RNase protection assay using 32P-labeled rat IGF-I and/or type I IGF receptor antisense RNA probes. Cultured in the absence or presence of FSH (100 ng/ml), whole ovarian dispersates (or isolated granulosa cells) displayed time-dependent (FSH-independent) decrements in the relative abundance of IGF-I transcripts apparent as early as 3 h after the onset of culture. No evidence of recovery was apparent by 96 h of culture. The apparent lack of an FSH effect did not reflect diminished biopotency as attested to by the ability of the hormone to promote time-dependent increments in the accumulation of progesterone. Importantly, the apparent decrease in ovarian IGF-I gene expression proved to be IGF-I specific in that type I IGF receptor transcripts displayed a substantial and sustained (for up to 96 h) FSH-independent increase beginning at the 24-h time point. At no point were IGF-II transcripts detected. The apparent decrease in the expression of IGF-I did not reflect the lack of extracellular matrix support in that neither laminin, collagen, nor whole serum supported sustained ovarian IGF-I gene expression. Treatment of whole ovarian dispersates with pharmacological concentrations of either insulin (1 micrograms/ml) or dexamethasone (10(-7) M) did not reverse the decline in IGF-I gene expression. Importantly, however, the combined application of both insulin and dexamethasone resulted in virtually complete preservation of IGF-I gene expression, the relative abundance of the corresponding transcripts proving uniform throughout. Taken together, these in vitro observations reveal irreversible (FSH-independent) decrements in ovarian IGF-I (but not type I IGF receptor) gene expression, the preservation of which required the concurrent provision of both insulin and dexamethasone.

Allograft replacement of the aortic valve versus the miniroot and valve.

Between February 1986 and April 1992, 27 patients ranging in age from 8 to 65 years (median, 18 years) underwent allograft replacement of the aortic valve with one death (operative mortality, 3.7%). The indications for operation were aortic regurgitation in 14 patients, aortic stenosis in 7, aortic stenosis/regurgitation in 4, and endocarditis in 2. Associated lesions included annuloectasia in 4 patients (1 with Marfan's syndrome), sinus of Valsalva aneurysm in 3, coronary artery disease in 4, and ventricular septal defect in 2. The freehand technique was used in 12 patients and the miniroot replacement in 15 patients. The postoperative transvalvular gradient was 17 mm Hg in freehand valves and 7 mm Hg in the miniroot valves. Initial postoperative study showed no or trivial aortic regurgitation in all 26 survivors except 1 freehand patient who had mild aortic regurgitation. At late study up to 6 years postoperatively all 10 of our available freehand patients showed progressive regurgitation, and it was severe in 4 patients. By contrast only half of the miniroot patients showed progressive regurgitation, and in none was it severe. Allograft replacement of the aortic valve can be done in children and adults with low mortality rate, short hospital stay, and excellent early function. Subsequent follow-up studies suggest that the miniroot replacements are superior to the freehand valves with lower transvalvular gradients, less valvular regurgitation, and delayed progression of valvular regurgitation.

Granulosa cell-derived insulin-like growth factor (IGF) binding proteins are inhibitory to IGF-I hormonal action. Evidence derived from the use of a truncated IGF-I analogue.

An increasing body of information now suggests that insulin-like growth factor (IGF) binding proteins (BPs) may serve as antigonadotropins at the level of the ovary. It is the objective of the present communication to evaluate the functional role of endogenous (granulosa cell-derived) IGFBPs by exploiting the unique properties of des(1-3)IGF-I, a naturally occurring IGF-I analogue characterized as a weak ligand of IGFBPs but not of type I IGF receptors. Given IGFBP-replete circumstances, des(1-3)IGF-I proved more potent (10-fold) than its intact counterpart in promoting the follicle stimulating hormone (FSH)-stimulated accumulation of progesterone by cultured rat granulosa cells. In contrast, des(1-3)IGF-I proved virtually equipotent to the unmodified principle under IGFBP-deplete circumstances. Taken together, these findings are in keeping with the notion and that the apparently enhanced potency of des(1-3)IGF-I (under IGFBP-replete conditions) is due to its diminished affinity for endogenously generated IGFBPs and that rat granulosa cell-derived IGFBPs are inhibitory to IGF (and thus inevitably to gonadotropin) hormonal action. Accordingly, the reported ability of gonadotropins to attenuate IGFBP release by granulosa cells may be designed to enhance the bioavailability of endogenously generated IGFs in the best interest of ovarian steroidogenesis.

Human intraovarian interleukin-1 (IL-1) system: highly compartmentalized and hormonally dependent regulation of the genes encoding IL-1, its receptor, and its receptor antagonist.

To delineate the scope of the human intraovarian IL-1 system we used a solution hybridization/RNase protection assay to test for expression of the genes encoding IL-1, its type I receptor (IL-1R), and its receptor antagonist (IL-1RA). IL-1 transcripts were not detected in whole ovarian material from days 4 or 12 of an unstimulated menstrual cycle but transcripts (IL-1 beta much greater than IL-11 alpha) were detected in preovulatory follicular aspirates from gonadotropin-stimulated cycles. Concurrently obtained peripheral monocytes did not contain IL-1 beta transcripts but macrophage-depleted follicular aspirates did, thus implicating the granulosa cells as the site of IL-1 expression. IL-1R transcripts were detected in RNA from whole ovaries and follicular aspirates but not in RNA from peripheral monocytes. IL-1RA transcripts were detected in whole ovarian material as well as in macrophage-free follicular aspirates. Cultured human granulosa and theca cells did not contain mRNA for IL-1 beta or IL-1RA but did contain mRNA for IL-1R. Treatment of cell cultures with forskolin (25 microM) induced IL-1 beta transcripts in granulosa but not theca cells. Forskolin also increased the basal levels of IL-1R transcripts in both granulosa and theca cells but did not induce IL-RA transcripts in either cell type. Taken together, these findings reveal the existence of a complete, highly compartmentalized, hormonally dependent intraovarian IL-1 system replete with ligands, receptor, and receptor antagonist.

Rat ovarian insulin-like growth factor binding protein-3: a growth hormone-dependent theca-interstitial cell-derived antigonadotropin.

To further the identification and characterization of insulin-like growth factor binding proteins at the level of the immature rat ovary, we have set out to study the ovarian expression, cellular localization, and hormonal regulation of the insulin-like growth factor binding protein (IGFBP)-3. To this end, use was made of a solution hybridization/RNAse protection assay wherein ovarian total RNA from immature (21-23 days old) female rats was hybridized with a 343 bases-long [32P]-labeled rat IGFBP-3 riboprobe. As in liver, a single protected fragment (315 bases-long) corresponding to IGFBP-3 transcripts was identified in whole ovarian material. Cellular localization studies revealed the IGFBP-3 gene to be exclusively expressed in the theca-interstitial rather than the granulosa cell compartment. To confirm presence and cellular distribution of the IGFBP-3 protein, media conditioned by cultured granulosa cells, theca-interstitial cells, and whole ovarian dispersates were subjected to Western Ligand Blotting. Importantly, media conditioned by cultured theca-interstitial (but not granulosa) cells displayed an IGFBP the size of rat IGFBP-3 (46kDa) as determined by comigration with a rat serum standard. A similarly-sized band was apparent in media conditioned by cultured whole ovarian dispersates reflecting in all likelihood the contribution of the theca-interstitial cell component. Significantly, deglycosylation of media conditioned by cultured theca-interstitial cells revealed the glycosylated nature of the 46kDa IGFBP species as judged by the apparent reduction in its molecular size to 35kDa. Similar alterations were noted in corresponding rat serum samples. Hypophysectomy of immature rats resulted in a modest but statistically insignificant decrease in the relative (densitometrically-quantified) abundance of ovarian IGFBP-3 transcripts, an effect further augmented by the systemic provision of either FSH or diethylstilbestrol (DES). In contrast, systemic treatment of hypophysectomized rats with GH produced a marked (3.2-fold) increase (P less than 0.05) in the steady state levels of ovarian (as well as hepatic) IGFBP-3 transcripts. However, the concurrent provision of either FSH or DES resulted in substantial (P less than 0.05) attenuation (78 and 57% inhibition, respectively) of the upregulatory GH effect. These findings document the highly compartmentalized expression of the IGFBP-3 gene at the level of the immature rat ovary, implicate the theca-interstitial cell as the sole source of its generation, reveal its pituitary dependence, and disclose its diametrically-opposed (indeed antagonistic) regulation by FSH (or estrogens) and GH.(ABSTRACT TRUNCATED AT 400 WORDS)