Viscosity and drag force involved in organelle transport: investigation of the fluctuation dissipation theorem.

We observed the motion of an organelle transported by motor proteins in cells using fluorescence microscopy. Particularly, among organelles, the mitochondria in PC12 cells were studied. A mitochondrion was dragged at a constant speed for several seconds without pausing. We investigated the fluctuation dissipation theorem for this constant drag motion by comparing it with the motion of Brownian beads that were incorporated into the cells by an electroporation method. We estimated the viscosity value inside cells from the diffusion coefficients of the beads. Then the viscosity value obtained by using the beads was found to be slightly lower than that obtained from the diffusion coefficient for the organelle motion via the Einstein relation. This discrepancy indicates the violation of the Einstein relation for the organelle motion.

Experience with CA 15.3 as a tumor marker in breast cancer.

CA 15.3, a new tumor marker, is a glycoprotein antigen produced in greater amounts by breast tumor cells. It can be quantitatively detected, circulating in human serum or plasma, using an immunoradiometric assay with monoclonal antibodies. In order to evaluate the usefulness of the method and to determine the cut-off for metastatic disease, the CA 15.3 levels were determined in 78 patients (5 patients with breast fibroadenoma and 73 patients with breast cancer). The conclusions of the study are that the CA 15.3 is a useful parameter in the management of patients in different stages of the disease: levels above 36 U/ml are suggestive of metastasis, and above 86 U/ml are indicative of them. On the other hand, CA 15.3 does not seem to be helpful in the pre-operative differential diagnosis of breast lumps.

Labeling of Salmonella typhimurium with iodine-131 to study phagocytic function in rats.

The present study describes a method for labeling Salmonella typhymurium with iodine-131 to evaluate both the morphological and the functional characteristics of the reticulo-endothelial system. A suspension containing 2 x 10(9) bacteria per ml was labeled with carrier-free Na131I without reductor, with a labeling yield of 46.5 +/- 3% and 3.5 +/- 1.3% of free Iodine-131. The biodistribution of the labeled bacteria in rats was studied with a large-field-of-view scintillation camera equiped with a pinhole collimator. Whole body images were obtained 15 and 30 minutes after intravenous injection of the labeled microorganisms. Images showed accumulation of bacteria in the liver and both normal and transplanted spleens of the animals. Autoradiographs of liver and spleen demonstrated labeled bacteria within the cells of the reticulo-endothelial system. The method described is easy to perform, has a good labeling yield and allows the functional evaluation of the reticulo-monophagocytic system, including transplanted spleens.

In vitro action of some disinfectants on Paracoccidioides brasiliensis yeast forms.

The fungicidal action of sodium hypochlorite (0.3, 1, 2.5, 5 and 10%); formaldehyde (2, 5, and 10%); and ethyl alcohol (70%) on yeast forms of Paracoccidioides brasiliensis Pb 18 and a newly-isolated Goiana strain was described. Contact between the fungus and the disinfectants was maintained for 1, 2, 24, 48 and 72 hours at room temperature. Viability was evaluated by the fluorescein diacetate-ethidium bromide treatment, culture in solid and liquid media (36 degrees C and 26 degrees C); yeast to mycelial germination at room temperature; and radiometric study of metabolic activity. All concentrations of disinfectants were found to be effective in inactivating Pb 18 and Goiana strains, except for the 1-hour contact with 2% formaldehyde, in which fluorescein diacetate-ethidium bromide treatment was found to reveal 40 and 27% of viable cells, respectively. The yeast to mycelial germination method was considered to reveal faster and similar results as compared to culture in solid and liquid media.

[Role of the process of endothelium formation on the wall of the aortic prosthesis in protection against infection. Experimental study in dogs]. / Rôle du processus de formation d'endothélium sur les parois de la prothèse de l'aorte dans la protection contre l'infection. Etude expérimentale sur des chiens.

An experimental canine model was developed in order to evaluate the role of endothelialization of aortic prostheses for protection against septicemic infection and the utilization of 131I radiolabeled bacteria for vascular experimentation. Two groups of dogs were submitted to insertion of a dacron prosthesis segment in the abdominal aorta with subsequent infusion of inert 131I radiolabeled bacteria. In the first group ("A"), formed by five dogs, the infusion was made 30 minutes after the insertion of the prosthesis and in the second group ("B"), formed by four dogs, in a new surgery performed 12 weeks after. Fragments of the animals' prostheses and aortas were collected after 30 minutes of septicemia and analyzed by scan electron microscopy and submitted to reading of radioactivity uptake by a well-type counter. The microscopy recognized a complete endothelialization of the prostheses of dogs of group "B" 12 weeks after their insertion. Statistical analysis comparing fragments of non-endothelialized prostheses, of endothelialized prostheses and aortas demonstrated that the lower radioactivity uptake of the endothelialized prostheses in relation to non-endothelialized ones was significant (p = 0.0143) and that there was no significant statistical difference in uptake in the aortas and in endothelialized prosthesis (p = 0.3173). It was, therefore, concluded that prosthesis endothelialization fully protected them against septicemic infection; bacteremia contaminated all the non-endothelialized prostheses; there was no bacterial adhesion in the endothelialized prostheses and the use of bacteria labeled with radioisotope 131I is appropriate for the study of infections in vascular prosthetic devices.

In vitro action of some disinfectants on Paracoccidioides brasiliensis yeast forms

Descreveu-se a acao fungicida do hipoclorito de sodio (0,3; 1; 2,5; 5 e 10 por cento); do formaldeido (em solucao aquosa a 2,5 e 10 por cento); e alcool etilico a 70,0 por cento sobre formas leveduriformes de 2 cepas de Paraccoccidioides brasiliensis: Pb 18 e cepa Goiana, recentemente isolada. A incubacao do fungo e desinfetantes foi realizada a temperatura ambiente por periodos de 1, 2, 24, 48 e 72 horas. A viabilidade foi avaliada pelo tratamento com diacetato de fluoresceina-brometo de etideo; pela cultura em meios solidos e liquidos a 36 graus Celsius e 26 graus Celsius; transformacao de levedura em micelio a temperatura ambiente; e estudo radiometrico da atividade metabolica. Todas as concentracoes de todos os desinfetantes estudados foram capazes de inativar ambas as cepas, exceto na incubacao com formaldeido a 2 por cento por 1 hora, em que o tratamento por diacetato de fluoresceina-brometo de etideo revelou 40 por cento e 27 por cento de celulas viaveis, respectivamente, para a cepa Pb 18 e Goiana. A transformacao de levedura em micelio foi considerada um metodo rapido, com resultados semelhantes ao cultivo em meios solidos e liquidos.