Toluene permeabilization differentially affects F- and P-type ATPase activities present in the plasma membrane of Streptococcus mutans

Streptococcus mutans membrane-bound P- and F-type ATPases are responsible for H+ extrusion from the cytoplasm thus keeping intracellular pH appropriate for cell metabolism. Toluene-permeabilized bacterial cells have long been used to study total membrane-bound ATPase activity, and to compare the properties of ATPase in situ with those in membrane-rich fractions. The aim of the present research was to determine if toluene permeabilization can significantly modify the activity of membrane-bound ATPase of both F-type and P-type. ATPase activity was assayed discontinuously by measuring phosphate release from ATP as substrate. Treatment of S. mutans membrane fractions with toluene reduced total ATPase activity by approximately 80% and did not allow differentiation between F- and P-type ATPase activities by use of the standard inhibitors vanadate (3 µM) and oligomycin (4 µg/mL). Transmission electron microscopy shows that, after S. mutans cells permeabilization with toluene, bacterial cell wall and plasma membrane are severely injured, causing cytoplasmic leakage. As a consequence, loss of cell viability and disruption of H+ extrusion were observed. These data suggest that treatment of S. mutans with toluene is an efficient method for cell disruption, but care should be taken in the interpretation of ATPase activity when toluene-permeabilized cells are used, because results may not reflect the real P- and F-type ATPase activities present in intact cell membranes. The mild conditions used for the preparation of membrane fractions may be more suitable to study specific ATPase activity in the presence of biological agents, since this method preserves ATPase selectivity for standard inhibitors.

Differential patterns of myosin Va expression during the ontogenesis of the rat hippocampus

Myosin Va is an actin-based, processive molecular motor protein highly enriched in the nervous tissue of vertebrates. It has been associated with processes of cellular motility, which include organelle transport and neurite outgrowth. The in vivo expression of myosin Va protein in the developing nervous system of mammals has not yet been reported. We describe here the immunolocalization of myosin Va in the developing rat hippocampus. Coronal sections of the embryonic and postnatal rat hippocampus were probed with an affinity-purified, polyclonal anti-myosin Va antibody. Myosin Va was localized in the cytoplasm of granule cells in the dentate gyrus and of pyramidal cells in Ammon's horn formation. Myosin Va expression changed during development, being higher in differentiating rather than already differentiated granule and pyramidal cells. Some of these cells presented a typical migratory profile, while others resembled neurons that were in the process of differentiation. Myosin Va was also transiently expressed in fibers present in the fimbria. Myosin Va was not detected in germinative matrices of the hippocampus proper or of the dentate gyrus. In conclusion, myosin Va expression in both granule and pyramidal cells showed both position and time dependency during hippocampal development, indicating that this motor protein is under developmental regulation.

A morphometric study of the lumbar spinous process in the Chinese population

Our goal was to analyze the anatomical parameters of the lumbar spine spinous process for an interspinous stabilization device designed for the Chinese population and to offer an anatomical basis for its clinical application. The posterior lumbar spines (T12-S1) of 52 adult cadavers were used for measuring the following: distance between two adjacent spinous processes (DB), distance across two adjacent spinous processes (DA), thickness of the central spinous processes (TC), thickness of the superior margin of the spinous processes (TS), thickness of the inferior margin of the spinous processes (TI), and height of the spinous processes (H). Variance and correlation analyses were conducted for these data, and the data met the normal distribution and homogeneity of variance. DB decreased gradually from L1-2 to L5-S1. DA increased from T12-L1 to L2-3 and then decreased from L2-3 to L4-5. The largest H in males was noted at L3 (25.45±5.96 mm), whereas for females the largest H was noted at L4 (18.71±4.50 mm). Usually, TS of the adjacent spinous process was lower than TI. Based on the anatomical parameters of the lumbar spinous processes obtained in this study, an “H”-shaped coronal plane (posterior view) was proposed as an interspinous stabilization device for the Chinese population. This study reports morphometric data of the lumbar spinous processes in the Chinese population, which provides an anatomical basis for future clinical applications.

Origanum vulgare L. essential oil as inhibitor of potentially toxigenic Aspergilli

Origanum vulgare L. essential oil has been known as an interesting source of antimicrobial compounds to be applied in food conservation. In this study, the effect of O. vulgare essential on the growth of A. flavus, A. parasiticus, A. fumigatus, A. terreus and A. ochraceus was assessed. The essential oil had a significant inhibitory effect on all assayed fungi. MIC was 0.6 µL.mL-1 for all fungi, while MFC was in the range of 1.25-2.5 µL.mL-1. The radial mycelial growth of A. flavus and A. parasiticus was strongly inhibited over 14 days at 0.6, 1.25 and 2.5 µL.mL-1 of oil in solid medium. The mycelial mass of all fungi was inhibited over 90% at 0.6 and 0.3 µL.mL-1 in liquid medium, while it was 100% at 1.25 µL.mL-1. The oil in a range of concentrations (0.6 to 2.5 µL.mL-1) was effective in inhibiting the viability and spores germination in a short time of exposure. The main morphological changes caused by the essential oil in A. parasiticus observed under light microscopy were absence of conidiation, leakage of cytoplasm, loss of pigmentation, and disrupted cell structure. These results demonstrated that O. vulgare essential oil produced a significant fungitoxic effect supporting its possible rational use as anti-mould compound in food conservation.

Biochemical and color changes of fresh-cut melon (Cucumis melo L. cv. Galia) treated with UV-C

The importance of minimally processed commodities in the retail groceries of most developed countries has been rising continuously during the last decades. Cantaloupe melon is used more than any other fruit in fresh-cut processing. Ultraviolet (UV) light has been extensively used to simulate biological stres in plants and for determining resistance mechanisms of plant tissues. In this study the effect of ultraviolet irradiation on some properties of fresh-cut cantalope melon was determined during storage. Freshly cut cantalope melons cubes treated with ultraviolet irradiation at the doses of 1, 2 or 3 min before storage, and then placed in a cold room at 5±1°C temperature and 85-90% RH. Hue angle values of control group is low compared to UV-C treated samples, whereas L values of is high. EL of UV treated samples higher than those of control group. Total soluble solids of fresh-cut melon samples in UC3 treatment increased during storage. The results indicate that UV-C treatments on fresh-cut cantaloupe melon cubes increased total soluble solids independently from water loss.

Biochemical changes and color properties of fresh-cut green bean (Phaseolus vulgaris L. cv.gina) treated with calcium chloride during storage

Calcium chloride is widely used in industries as a firming agent, and also to extend shelf-life of vegetables. The aim of this study was to determine, the effect of different doses of calcium chloride on biochemical and color properties of fresh-cut green bean. Fresh-cut green beans were dipped for 90 seconds in 0.5%, 1%, 2% and 3% solution of calcium chloride at 25°C. The fresh-cut green bean samples were packaged in polystyrene foam dishes, wrapped with stretch film and stored in a cold room at 5±1°C temperature and 85-90% RH. Calcium chloride treatments did not retain the green color of samples. Whiteness index, browning index and total color difference (ΔE) values of CaCl2 treated samples were high. Saturation index and hue angle were low compared to the control, especially at higher doses of CaCl2. Polyphenol oxidase (PPO) enzyme activity in samples treated with CaCl2 at 3% doses, was low at the 7th days of storage than with other treatments. Fructose and sucrose content of samples increased in all treatment groups whereas glucose level decreased during the first 4th days of storage.

Building-up influence: post-war industrialization in the State of Minas Gerais, Brazil

This paper examines the post-War industrialization process in the Brazilian State of Minas Gerais, focusing on one of its desirable outcomes, namely the capacity to generate growth through the impact of strong input-output linkages. This process is placed into historical perspective considering the ideas that permeate the economic development debate throughout the period of analysis. Changes in the regional economic structure are assessed through the use of three input-output tables for the years of 1953, 1980 and 1995. By adopting the fields of influence methodology as the analytical core, it is shown that the efforts towards the creation of a more integrated regional economy have generated stronger influence of the targeted sectors (metal products, transportation equipment, chemical, and services). However, structural changes also contributed to strengthen leakage in the system originated in traditional economic activities.