Application of Saccharomyces cerevisiae var. boulardii in food processing: a review.

Probiotics are increasingly being added to food in order to develop products with health-promoting properties. Particularly, Saccharomyces cereviceae var. boulardii yeast is recently being investigated like a starting-culture for development of functional and probiotic foods. Although the literature is abundant on the beneficial effects of S. boulardii on health, slight information is available on the effects of supplementing this probiotic to food systems. The aim of this paper is to examine the applications of S. boulardii to different food matrices and its implication in food processing (stability, sensorial properties and other technological implications) and the concomitant effects on nutrition and health.

Participation of PLA2 and PLC in DhL-induced activation of Rhinella arenarum oocytes.

Rhinella arenarum oocytes can be artificially activated, a process known as parthenogenesis, by a sesquiterpenic lactone of the guaianolide group, dehydroleucodine (DhL). Transient increases in the concentration of cytosolic Ca2+ are essential to trigger egg activation events. In this sense, the 1-4-5 inositol triphosphate receptors (IP3R) seem to be involved in the Ca2+ transient release induced by DhL in this species. We analyzed the involvement of phosphoinositide metabolism, especially the participation of phospholipase A2 (PLA2) and phospholipase C (PLC) in DhL-induced activation. Different doses of quinacrine, aristolochic acid (ATA) (PLA2 inhibitors) or neomycin, an antibiotic that binds to PIP2, thus preventing its hydrolysis, were used in mature Rhinella arenarum oocytes. In order to assay the participation of PI-PLC and PC- PLC we used U73122, a competitive inhibitor of PI-PLC dependent events and D609, an inhibitor of PC-PLC. We found that PLA2 inhibits quinacrine more effectively than ATA. This difference could be explained by the fact that quinacrine is not a specific inhibitor for PLA2 while ATA is specific for this enzyme. With respect to the participation of PLC, a higher decrease in oocyte activation was detected when cells were exposed to neomycin. Inhibition of PC-PLC with D609 and IP-PLC with U73122 indicated that the last PLC has a significant participation in the effect of DhL-induced activation. Results would indicate that DhL induces activation of in vitro matured oocytes of Rhinella arenarum by activation of IP-PLC, which in turn may induce IP3 formation which produces Ca2+ release.

Chemical activation in Rhinella arenarum oocytes: effect of dehydroleucodine (DhL) and its hydrogenated derivative (2H-DhL).

Mature oocytes are arrested in metaphase II due to the presence of high levels of active maturation promoting factor (MPF). After fertilization, active MPF levels decline abruptly, enabling oocytes to complete meiosis II. One of the first and universal events of oocyte activation is an increase in cytosolic Ca2+ that would be responsible for MPF inactivation. Mature oocytes can also be activated by parthenogenetic activation. The aims of this work are to test the ability of dehydroleucodine (DhL) and its hydrogenated derivative 11,13-dihydro-dehydroleucodine (2H-DhL) to induce chemical activation in amphibian oocytes and to study the participation of calcium in the process. Results indicated that DhL and 2H-DhL induced oocyte activation in a dose-dependent manner. After 90 min of treatment, DhL 36 µM was able to induce 95% activation, while 2H-DhL 36 µM was less active, with only 40% activation. Our results suggest that DhL induced the inhibition of MPF activity, probably by an increase in intracellular Ca2+ concentration. Extracellular Ca2+ would not be significant, although Ca2+ release from intracellular stores is critical. In this sense, IP3Rs and RyRs were involved in the Ca2+ transient induced by lactones. In this species, RyRs appears to be the largest contributor to Ca2+ release in DhL-induced activation. Although more studies are needed on the mechanism of action through which these lactones induce oocyte activation in Rhinella arenarum, the results of this research provide interesting perspectives for the use of these lactones as chemical activators in in vitro fertilization and cloning.

Immunocytochemical characteristics of elbow, knee and ankle muscles of the five-toed jerboa (Allactaga elater).

Biochemical adaptations of limb myofibres to intensive bipedal hopping were investigated using the five-toed jerboa Allactaga elater as a model in comparison with the rat. Immunofluorescence methods included immunoreactivity to anti-fast and anti-slow MHC and troponin I. There is no specialization of triceps caput mediale for postural function in the minute non-locomotor forelimbs, unlike quadruped mammals. The various elbow extensor heads and the flexor muscles are alike with regard to fibre type population and cross-sectional areas of each type of fibre. The extensor muscle in the elongated hindlimbs of the five-toed jerboa, at both the knee and the ankle joints, differ from each other extensively. One head, made up of an extremely high percentage of type I, fatigue-resistant fibres, is suited to postural function. Two extensor heads at each joint contain a very high percentage of type IIB fibres (having the greatest maximal velocity of contraction) and are able to produce the powerful acceleration needed to trigger the leap. The relative cross-sectional areas of the myofibres are characteristic of hopping locomotion: predominance in number of one type of myofibre in a muscle accompanies greater cross-sectional area, which increases muscle efficiency in either postural or accelerative function of the muscle.

Nuclear maturation inducers in Bufo arenarum oocytes.

The present study analyses the effect of dihydrotestosterone (DHT) and mammalian insulin on the nuclear maturation of Bufo arenarum oocytes under in vitro conditions. The response of fully grown follicle oocytes to DHT, shown by germinal vesicle breakdown (GVBD), occurred in a manner dependent on dose, time and sexual cycle period. The highest oocyte sensitivity to the hormone appeared during the breeding period, a fact evinced by high GVBD percentages after short incubation periods and at a low hormone concentrations. Insulin also proved effective in inducing nuclear maturation, although its action was only visible at high concentrations and after a long incubation period. The combination of insulin and steroid hormones (DHT or progesterone), both at subliminal doses, caused a noticeable potentiating synergism, resulting in a rapid and important increase in GVBD. Another effect of insulin was the acquisition by oocytes of steroid sensitivity during folliculogenesis.

Function and cytochemical characteristics of postural limb muscles of the rhesus monkey: A telemetered EMG and immunofluorescence study.

Using telemetered electromyography and immunocytochemical fibre typing (of both fresh frozen and preserved specimens), the present paper demonstrates clearly that at the elbow, knee, and ankle joints, the rhesus monkey (Macaca mulatta) is endowed with one extensor-muscle head specialized for posture. These postural heads are distinguished by (a) recruitment at low levels to maintain joint position against the effect of gravity, (b) recruitment near maximum levels during walking, and (c) high content and relatively large size of slow, fatigue-resistant (type I) muscle fibres. The nonpostural heads of the investigated muscles were recruited at levels correlated to the strenuousness of the effort and are notable by the small percentage and size of slow muscle fibres. The postmortem stability of the structural properties of myosin makes immunocytochemical fibre typing suitable for the study of preserved cadavers.

Design, characterization and testing of tRNA3Lys-based hammerhead ribozymes.

A hammerhead ribozyme targeted against the HIV-1 env coding region was expressed as part of the anticodon loop of human tRNA3Lys without sacrificing tRNA stability or ribozyme catalytic activity. These tRNA-ribozymes were isolated from a library which was designed to contain linkers (sequences connecting the ribozyme to the anticodon loop) of random sequence and variable length. The ribozyme target site was provided in cis during selection and in trans during subsequent characterization. tRNA-ribozymes that possessed ideal combinations of linkers were expected to recognize the cis target site more freely and undergo cleavage. The cleaved molecules were isolated, cloned and characterized. Active tRNA-ribozymes were identified and the structural features conducive to cleavage were defined. The selected tRNA-ribozymes were stable, possessed cleavage rates lower or similar to the linear hammerhead ribozyme, and could be transcribed by an extract containing RNA polymerase III. Retroviral vectors expressing tRNA-ribozymes were tested in a human CD4+ T cell line and were shown to inhibit HIV-1 replication. These tRNA3Lys-based hammerhead ribozymes should therefore prove to be valuable for both basic and applied research. Special application is sought in HIV-1 or HIV-2 gene therapy.

RNA-polymerase III-driven expression cassettes in human gene therapy.

tRNAs, U6 small nuclear RNA and adenovirus virus associated RNAs are transcribed by RNA polymerase III. These RNAs are expressed in high levels and possess increased stability due to their small, compact size. Promoters expressing these RNAs have therefore been used to express a variety of interfering RNAs, including ribozymes, antisense RNAs and sense or decoy RNAs. Intracellular expression and activity of RNA polymerase III-driven interfering RNAs have been demonstrated in several studies. This review will summarize the design and application of RNA polymerase III-driven expression cassettes in human gene therapy using retroviral vectors as the primary method of gene delivery.

Developmental changes in the fibre composition of elbow, knee, and ankle extensor muscles in cercopithecid monkeys.

Developmental changes of muscle fibre composition in the various heads of the elbow, knee, and ankle extensors have been studied in three genera of cercopithecid monkeys. In order to circumvent the technical hindrances of usual histoenzymological procedures (fresh muscles need to be frozen at once at -80 degrees C), immunofluorescence methods were used and technical adjustments were successfully carried out to make the study of formaldehyde-preserved muscles possible. Clear responses to antibodies against adult fast myosin in newborn macaques demonstrated that, at birth, adult myosins have already replaced the fetal isoforms, thus providing a reliable marker for the study of postnatal evolution of the muscle fibre composition. For each one of the three joints, from birth to adulthood, the percentage of slow, fatigue-resistant fibres increases only in that head of the extensor muscle groups which is specialized in maintaining posture by counteracting gravity (the 'postural' head). Hence, the question is raised of the relationships between such cytological evolution, developmental changes in postural behaviour, and body weight increase.

Broncoscopia en el diagnóstico del cáncer broncogénico / Broncoscopy in the diagnosis of bronchogenic carcinoma

Se presentan los resultados obtenidos con el estudio broncoscópico de 637 pacientes con carcinoma broncogénico. Las imagenes broncoscópicas observadas fueron: masa tumoral, estenosis bronquial, hemorragia y mucosa de aspecto normal. El pulmón derecho fue el más afectado con 203 casos, mientras que en el izquierdo se presentaron 11. En la tráquea se encontraron tumoraciones en 31 enfermos. Se señalan los sitios de las vías aéreas donde se localizaron estas alteraciones y su frecuencia. El estudio citológico del lavado y cepillado bronquial mediante broncoscopia evidenció 181 casos con células neoplásicas, cifra que se incrmentó en un 3.4 por ciento en muestras de concentrado de expectoración en líquido de carbowax en los días subsecuentes a la broncoscopia. Se efectuaron 348 biopsias bronquialeas durante el estudio broncoscópico, encontrando que el tipo histológico de cáncer más frecuente correspondió al epidermoide con 165 casos, seguido del adenocarcinoma con 143. Se considera que la fibrobroncoscopia es un método seguro y que debe efectuarse en los pacientes con sospecha clínica de padecer cáncer pulmonar

[Levogram in congenital cardiopathies. Its practical usefulness in newborn infants and small children given its low risk]. / El levograma en las cardiopatías congénitas. Su utilidad práctica en recién nacidos y niños pequeños por su bajo riesgo.

Four hundred and fifty six angiocardiograms were studied to evaluate the validity of levogram for diagnosis. They were separated in three groups: 1. Right angiocardiogram with a levogram. 2. Right angiocardiogram and left angiocardiogram. 3. Left angiocardiogram only. The distribution according to age, and type of heart disease is shown. The correlation between levogram and left angiocardiogram was looked for. The findings at survey were correlated with the angiocardiographic diagnosis and a high correlation was established. The usefulness of this procedure to diagnose specific type of heart disease in the small child is emphasized.