Sucrose-phosphate phosphatase from Anabaena sp. strain PCC 7120: isolation of the protein and gene revealed significant structural differences from the higher-plant enzyme.

The present study describes the first isolation and characterization of a prokaryotic protein and gene for sucrose-phosphate phosphatase (SPP), the enzyme that catalyzes the terminal step in sucrose synthesis. For gene isolation, a 2,015-bp DNA fragment containing an open reading frame with about 31% amino acid identity to Synechocystis SPS was amplified from Anabaena sp. PCC 7120 DNA. Surprisingly, expression of the putative gene in Escherichia coli demonstrated that it encoded an SPP protein. The expressed protein cross-reacted with antibodies against the native form of Anabaena SPP and its biochemical properties were identical to those of the enzyme purified from the cyanobacterial cells. Comparisons of the Anabaena SPP with the higher-plant enzyme revealed important differences in the C-terminal region, molecular mass, subunit composition and immunoreactivity. Nevertheless, two conserved motifs, including four invariant aspartate residues similar to those found in members of the phosphohydrolase superfamily, were identified in the Anabaena SPP deduced amino acid sequence.

[Splenic artery aneurysm. Two successfully operated cases]. / Gli aneurismi dell'arteria splenica. Su due casi operati con successo.

The splenic artery aneurysms usually have an asymptomatic course; their rupture is associated with high mortality. The clinical history and the treatment of two patients with splenic artery aneurysm are reported. Of the two cases, one was asymptomatic, diagnosed with upper abdomen ultrasound, the other one was treated as an emergency because presented with shock. Both patients underwent surgical procedure; ligature of the splenic artery and splenectomy were performed. Early diagnosis is important in these lesions because the progressive enlargement and eventual rupture are the natural history. Ultrasonography (US) and computed tomography (TC) are very helpful as diagnostic tools, however angiography represents the method of choice since it could be the first step of the embolization treatment. If the diameter is more than 2.5 cm surgical treatment must be performed. Ruptures are treated with emergency operations, when possible. In high-risk patients non operative management by selective embolization may be a suitable alternative.

[Presence of Mycoplasma in laboratory cell cultures from Cordoba, Argentina]. / Presencia de micoplasmas en cultivos celulares en laboratorios de la ciudad de Córdoba, Argentina.

In this paper we determined the prevalence of mycoplasma contamination in 17 cell lines. Eighty per cent of the laboratories that currently use cell culture techniques participated in this study. Hoechst 33258 dye was used to detect mycoplasma contamination. The relationship between culture maintenance conditions and the presence of mycoplasma were analyzed, considering the use of antibiotics in the culture media, fetal calf serum (FCS) quality, culture media processing, use of disponsable labware, type of laminar flow cabinet, quantity of operators, and cell culture system. Thirty-five per cent of the analyzed cell lines showed mycoplasma contamination. Those lines belonged to 2 of the 8 surveyed laboratories. When confronting the working conditions versus mycoplasma contamination, 66% of the laboratories that employ non-certified FCS or reuse their labware, show mycoplasma contamination. Mycoplasma presence was found in 50% of the laboratories that use closed culture system, or more than one operator. Laboratories that process their culture media or that include antibiotic in the growing media, show a 40% contamination. The results obtained help to establish working conditions necessary to avoid introducing or spreading the microorganism.

Presencia de micoplasmas en cultivos celulares en laboratorios de la ciudad de Córdoba, Argentina / Mycoplasma presence in cell cultures in laboratories of Cordoba, Argentina

En este trabajo se determinó la presencia de micoplasmas en 17 líneas celulares continuas, provenientes de 8 laboratorios de cultivos celulares que corresponden al 80 por ciento de los que utilizan cultivos celulares en la ciudad de Córdoba. La contaminación fue determinada con el reactivo fluorescente Hoechst 33258. Se analizó la relación entre las condiciones de trabajo de cada laboratorio con la contaminación de sus líneas celulares. Las variables de trabajo estudiadas fueron: uso de antibióticos, calidad del suero fetal bovino (SFB), procesamiento del medio de cultivo, uso de materiales descartables, tipo de cabina de flujo laminar, sistema de crecimiento de cultivo y número de operadores. El índice de contaminación por micoplasmas fue 35 por ciento. De los laboratorios evaluados, sólo dos presentaron contaminación. Al evaluar las condiciones de trabajo en cada laboratorio y relacionarlas con la contaminación por micoplasmas, surgió 66 por ciento de infección entre los laboratorios que utilizaban SFB no certificados o reutilizaban el material de laboratorio; 50 por ciento, entre los laboratorios que contaban con sistemas cerrados de crecimiento celular o con un número de operadores mayor a uno; y 40 por ciento en aquéllos que incluían el procesamiento de los medios de cultivo (filtración o dilución) o utilizaban antibióticos en el medio de crecimiento. Los resultados obtenidos de este análisis contribuyen a sugerir condiciones de trabajo que tiendan a disminuir el riesgo de contaminación (AU)

Presencia de micoplasmas en cultivos celulares en laboratorios de la ciudad de Córdoba, Argentina / Mycoplasma presence in cell cultures in laboratories of Cordoba, Argentina

En este trabajo se determinó la presencia de micoplasmas en 17 líneas celulares continuas, provenientes de 8 laboratorios de cultivos celulares que corresponden al 80 por ciento de los que utilizan cultivos celulares en la ciudad de Córdoba. La contaminación fue determinada con el reactivo fluorescente Hoechst 33258. Se analizó la relación entre las condiciones de trabajo de cada laboratorio con la contaminación de sus líneas celulares. Las variables de trabajo estudiadas fueron: uso de antibióticos, calidad del suero fetal bovino (SFB), procesamiento del medio de cultivo, uso de materiales descartables, tipo de cabina de flujo laminar, sistema de crecimiento de cultivo y número de operadores. El índice de contaminación por micoplasmas fue 35 por ciento. De los laboratorios evaluados, sólo dos presentaron contaminación. Al evaluar las condiciones de trabajo en cada laboratorio y relacionarlas con la contaminación por micoplasmas, surgió 66 por ciento de infección entre los laboratorios que utilizaban SFB no certificados o reutilizaban el material de laboratorio; 50 por ciento, entre los laboratorios que contaban con sistemas cerrados de crecimiento celular o con un número de operadores mayor a uno; y 40 por ciento en aquéllos que incluían el procesamiento de los medios de cultivo (filtración o dilución) o utilizaban antibióticos en el medio de crecimiento. Los resultados obtenidos de este análisis contribuyen a sugerir condiciones de trabajo que tiendan a disminuir el riesgo de contaminación

[Deep venous thrombosis in a patient with superficial thrombophlebitis. Apropos of 2 cases]. / Trombosi venosa profonda in paziente con tromboflebite superficiale. A proposito di due casi.

Deep vein thrombosis in patient with superficial thrombophlebitis of the leg. Description of two cases. Two clinical cases already observed at our Department of Vascular Surgery, regarding patients with superficial thrombophlebitis (STF) associated with a deep venous thrombosis (DVT) are presented. The diagnosis of DVT was achieved exclusively on the basis of the duplex scanner because the two patients were completely asymptomatic. The chance that symptoms and clinical signs of STF cover a DVT is a possibility that should always be considered. For this reason every STF should be further investigated with duplex scanner in order to exclude that the deep venous circulation get involved at the same time. The DVT in these cases is generally due to the spreading of the thrombosis process from the perforating branches. The chance of a multiple onset of thrombosis in the superficial and deep circulation represents anyway an hypothesis worth of studying.

Culture amplification in human colon adenocarcinoma cell line (CaCo-2) combined with an ELISA as a supplementary assay for accurate diagnosis of rotavirus.

Culture amplification in colon adenocarcinoma cell line (CaCo-2) combined with enzyme immunoassay (Pathfinder ELISA) was developed as a supplementary tool for rotavirus diagnosis. One hundred and thirty stools in which results by polyacrylamide gel electrophoresis (PAGE) were in agreement with those obtained by ELISA were amplified in the CaCo-2 cell line. After the first passage 100% specimens were revealed as positive by ELISA. This result was confirmed by PAGE and direct electron microscopy (EM) which increased the rates of rotavirus detection up to 100% after the third and fifth cell passages, respectively. All of the amplified negative stools were confirmed as negative. Among discordant results, three of the eight specimens positive by ELISA but negative by PAGE were confirmed as true positive after the third cell passage. False positive ELISA results could be discarded when the samples were culture amplified and retested by the same ELISA. Using the CaCo-2 amplification-ELISA as supplementary assay, sensitivity and specificity were 1.000 and 0.953 for ELISA and 0.917 and 1.000 for PAGE, respectively. The combined CaCo-2 cell line amplification-immunoassay method proved to be suitable both to evaluate increase in sensitivity of newly developed rotavirus assays and for rotaviral amplification before antigen assays.

[Clinico-statistical evaluation of oral hygiene in school-age patients with special reference to 1000 cases referred to orthodontic treatment and under observation at the Torino University Dental Clinic]. / Valutazione clinico-statistica dell'igiene orale in pazienti in età scolare con particolare riferimento a 1000 casi motivati al trattamento ortodontico o in osservazione presso la Clinica Odontoiatrica Università di Torino.

The importance of dental hygiene and the insufficient sensibilization to the problem, statistically proved, are underlined.