Set-up of a method using LC-UV to assay mometasone furoate in pharmaceutical dosage forms

A reliable method using LC-UV to assay mometasone furoate (MF) in creams or nasal sprays using the same chromatographic conditions was set up. Methanol:water 80:20 (v/v) (1.0 mL min-1) was used as mobile phase. MF was detected at 248 nm and analyzed in a concentration range from 1.0 to 20.0 µg mL-1. The method provided acceptable theoretical plates, peak simmetry, peak tailing factor and peak resolution a short run (5 min). The method showed specificity, good linearity (r = 0.9999) and the quantification limit was 0.379 µg mL-1. Furthermore, the method was precise (RSD < 2.0%), accurate (recovery > 97%) and robust.

Hydrozoa e Scyphozoa existentes no instituto paulista de oceanografia

Continuing the inventory of the Coelenterate Fauna of the Brazilian coast, 23 species of Hydrozoa and a Scyphomedusa are recorded from different points of the coast aproximately between 22º and 24º lat. S. Of these, 14 species were already known from other points of the coast and the remaining 10 were unknown from this latitude. Two species are here listed as new, they are: Halocordyle fragHis and Calicella gabriellae. 1. - Halocordyle fragilis, Stechow's (1923) correction of the generic name Pennaria has been followed, shows remarkable distinctive specific characters such as: alternate distribution of the cladii which do not lie all in the same plane, they are inserted at random on the hydrocaulus with a tendency towards a spiral dexiotropic distribution, sparse distribution of the pedicels which are alternate and not all located on the same side of the hydrocladium, and, finally, the long, wiry aspect of the colony which is as brittle as light glass. This characteristics are not sufficient, to my belief, to establish a new genus, since the polyps and the gonophores are entirely of the Halocordyle type, but they confer upon the species a very peculiar habitus. 2. - Calicella gabriellae forms erect hydrocauli which are extremely soft and flexuose, with hydrothecate and intermediate internodes. The thecae are deep, cylindrical, with a conical base, there are 10-12 long and narrow opercular valves. The gonangia are small, conical, with a short distal digitiform process. The included sporosac exhibits maturing ova. Dynamena heteroclonta described by Jarvis (1922) as a new species, is here considered as a new form of the very variable species Dynamena quadridentata. Schizotricha billardi nom. nov., is specifically separated from Sch. diaphana (Heller) to which it had been identified up to know. The Brazilian colonies agree with the French Somaliland specimens described by Billard (1904) and they are reunited in a new species named after the French worker. The main difference, between the two mentioned species are: shape and size of the gonothecae, of the thecae and of the nematophores. A description is given of all the species not previously recorded from the Brazilian coast as well as biological observations on Olindias sambaquiensis whose shoals during winter months are very large and may entangle partially the fishermen's nets. Its feeding and swimming habits are described and the species proves to be an excellent laboratory animal both for class and research purposes, for it easily endures long transportation (even up to 12 hours during summer months) as well as unoxygenized acquaria and nevertheless greadily feeds on any animal food.

Argentimetric assay of ranitidine in bulk drug and in dosage forms

Two simple, rapid and cost-effective methods based on titrimetric and spectrophotometric techniques are described for the assay of RNH in bulk drug and in dosage forms using silver nitrate, mercury(II)thiocyanate and iron(III)nitrate as reagents. In titrimetry, an aqueous solution of RNH is treated with measured excess of silver nitrate in HNO3 medium, followed by determination of unreacted silver nitrate by Volhard method using iron(III) alum indicator. Spectrophotometric method involve the addition a known excess of mercury(II)thiocyanate and iron(III)nitrate to RNH, followed by the measurement of the absorbance of iron(III)thiocyante complex at 470 nm. Titrimetric method is applicable over 4-30 mg range and the reaction stoichiometry is found to be 1:1 (RNH: AgNO3). In the spectrophotometric method, the absorbance is found to increase linearly with concentration of RNH which is corroborated by the correlation coefficient of 0.9959. The system obey Beer's law for 5-70 µg mL-1. The calculated apparent molar absorptivity and sandell sensitivity values are found to be 3.27 ´ 10³ L mol-1 cm-1, 0.107 µg cm-2 respectively. The limits of detection and quantification are also reported for the spectrophotometric method. Intra-day and inter-day precision and accuracy of the methods were evaluated as per ICH guidelines. The methods were successfully applied to the assay of RNH in formulations and the results were compared with those of a reference method by applying Student's t and F-tests. No interference was observed from common pharmaceutical excipients. The accuracy of the methods was further ascertained by performing recovery tests by standard addition method.

Development and validation of spectroscopic methods for simultaneous estimation and dissolution of ofloxacin and ornidazole in tablet dosage forms

The aim of this work was to develop and validate simple, accurate and precise spectroscopic methods (multicomponent, dual wavelength and simultaneous equations) for the simultaneous estimation and dissolution testing of ofloxacin and ornidazole tablet dosage forms. The medium of dissolution used was 900 ml of 0.01N HCl, using a paddle apparatus at a stirring rate of 50 rpm. The drug release was evaluated by developed and validated spectroscopic methods. Ofloxacin and ornidazole showed 293.4 and 319.6nm as λmax in 0.01N HCl. The methods were validated to meet requirements for a global regulatory filing. The validation included linearity, precision and accuracy. In addition, recovery studies and dissolution studies of three different tablets were compared and the results obtained show no significant difference among products.

Terrain forms and spatial variability of soil properties in an area cultivatedwith citrus

The technique of precision agriculture and soil-landscape allows delimiting areas for localized management, allowing a localized application of agricultural inputs and thereby may contribute to preservation of natural resources. Therefore, the objective of this work was to characterize the spatial variability of chemical properties and clay content in the context of soil-landscape relationship in a Latosol (Oxisol) under cultivation of citrus. Soil samples were collected at a depth of 0.0-0.2 m in an area of 83.5 ha planted with citrus, as a 50-m intervals grid, with 129 points in concave terrain and 206 points in flat terrain, totaling 335 points. Values for the variables that express the chemical characteristics and clay content of soil properties were analyzed with descriptive statistics and geostatistical modeling of semivariograms for making maps of kriging. The values of range and kriging maps indicated higher variability in the shape of concave topography (top segment) compared with the shape of flat topography (slope and hillside segments below). The identification of different forms of terrain proved to be efficient in understanding the spatial variability of chemical properties and clay content of soil under cultivation of citrus.

Immunohistochemical study of genital and extragenital forms of canine transmissible venereal tumor in Brazil

Aiming to provide insight and discussing the problems related to the diagnosis and differential diagnosis of canine transmissible venereal tumor (CTVT), especially in its extragenital form, immunohistochemical evaluation was performed and a comparison was established by analysis of the microscopic appearance of 10 genital CTVTs and 13 exclusively extragenital CTVTs previously diagnosed by cytology and histopathology. CTVTs samples were incubated with biotinylated antibodies raised against specific membrane (anti-macrophage) and cytoplasmic antigens (anti-lysozyme, anti-S-100 protein, anti-vimentin and anti-CD18) and subsequently developed using streptavidin-biotin peroxidase and streptavidin-biotin-alkaline phosphatase methods. A strong reactivity with the anti-vimentin antibody was found in 100% of the tumors tested (22/22). No reactivity was found for the anti-lysozyme, anti-macrophage, anti-S-100 protein and anti-CD18. No histopathological or immunoreactivity differences between genital and extragenital CTVTs were found. These findings do not corroborate the hypothesis of histiocytic origin of CTVT (no reactivity to anti-lysozyme, anti-macrophage and anti-CD 18 antibodies). In addition, the antibody panel used is useful to narrow the differential diagnosis for lymphomas, histiocytic tumors, amelanotic melanomas, and poorly differentiated epithelial neoplasias, among others.

Multiple forms of cotyledonary b-galactosidases from Vigna unguiculata quiescent seeds

Cotyledonary b-galactosidases were isolated and partially purified from Pitiúba cowpea (Vigna unguiculata (L.) Walp.) quiescent seeds. The purification steps consisted of precipitation of the crude extract with ammonium sulphate in the range of 20-60% saturation, acid precipitation, DEAE-Sephadex ion-exchange chromatography and Lactosyl-Sepharose affinity chromatography. This purification process gave rise to three b-galactosidases-rich fractions: b-gal I, b-gal II and b-gal III, which were purified about 5, 509, and 62 fold, respectively. They reached maximal enzyme activity at different pH ranges: 3.5-4.5 for b-gal I, 3.0-3.5 for b-gal II, and 3.0-4.0 for b-gal III. Their maximal activities were reached when the temperature of the assay medium was 60° C, and preincubation of the enzymes at different temperatures has shown that they were heat-stable up to 50° C. There were no significant differences among the partially purified enzymes as far as their response to the different effectors tested, except for Mn2+ and EDTA, which affected differently b-gal I, b-gal II, and b-gal III. They were slightly affected by Mg2+, Ca2+, Zn2+, Co2+, tartarate, molybdate, glucose, and lactose, strongly inhibited by Cu2+ and galactose, and inactivated by Hg2+. These chemical and physical properties are similar to the ones found for other plant b-galactosidases. Although through this process of purification three isoforms of this enzyme were obtained, isoelectric focusing in polyacrylamide slab gel of these enzyme-proteins suggest that cotyledons of Pitiúba cowpea quiescent seeds possess four isoforms of b-galactosidases.

Oral tolerance induction with altered forms of ovalbumin

As a T cell-dependent phenomenon, oral tolerance is not expected to depend necessarily on native configuration of antigens. We investigated the induction of oral tolerance with modified ovalbumin (Ova). Oral administration of heat-denatured (HD-Ova) and cyanogen bromide-degraded ovalbumin was less effective than native Ova in inducing oral tolerance in B6D2F1 mice. HD-Ova was effective in suppressing delayed-type hypersensitivity (DTH) reactions but did not suppress specific antibody formation. Injection of Ova directly into the stomach, but not into the ileum or cecum, suppressed subsequent immunization to DTH reactions. Gavage with protease inhibitors (aprotinin or ovomucoid) before gavage with Ova was ineffective in blocking tolerance induction. Treatment with hydroxyurea to destroy cycling cells 24 h before gavage with Ova blocked oral tolerance induction and also the possibility to passively transfer tolerance to naive recipients with the serum of mice gavaged with Ova 1 h before. The implications of these findings about oral tolerance induction are discussed

Influence of exercise on the activity and the distribution between free and bound forms of glycolytic and associated enzymes in tissues of horse mackerel

The effects of short-term burst (5 min at 1.8 m/s) swimming and long-term cruiser (60 min at 1.2 m/s) swimming on maximal enzyme activities and enzyme distribution between free and bound states were assessed for nine glycolytic and associated enzymes in tissues of horse mackerel, Trachurus mediterraneus ponticus. The effects of exercise were greatest in white muscle. The activities of phosphofructokinase (PFK), pyruvate kinase (PK), fructose-1,6-bisphosphatase (FBPase), and phosphoglucomutase (PGM) all decreased to 47, 37, 37 and 67%, respectively, during 60-min exercise and all enzymes except phosphoglucoisomerase (PGI) and PGM showed a change in the extent of binding to subcellular particulate fractions during exercise. In red muscle, exercise affected the activities of PGI, FBPase, PFK, and lactate dehydrogenase (LDH) and altered percent binding of only PK and LDH. In liver, exercise increased the PK activity 2.3-fold and reduced PGI 1.7-fold only after 5 min of exercise but altered the percent binding of seven enzymes. Fewer effects were seen in brain, with changes in the activities of aldolase and PGM and in percent binding of hexokinase, PFK and PK. Changes in enzyme activities and in binding interactions with subcellular particulate matter appear to support the altered demands of tissue energy metabolism during exercise.

Circulating forms of parathyroid hormone detected with an immunofluorometric assay in patients with primary hyperparathyroidism and in hyperparathyroidism secondary to chronic renal failure

In patients with uremia, intact parathyroid hormone (PTH) measurement appears to overestimate the biologically active hormone in circulation. The recent description of the accumulation in these patients of a non-intact PTH form measured by the standard immunometric assays, re-opened the question. In this study we submitted serum samples from 7 patients with primary hyperparathyroidism (PHP) and from 10 patients with hyperparathyroidism secondary to chronic renal failure (SHP) to preparative HPLC in order to discriminate the molecular forms measured by our currently used immunofluorometric assay for intact PTH. The elution profile obtained with the HPLC system showed two clearly defined peaks, the first one corresponding to a lower molecular weight form, and the second to the intact PTH (1-84) form. In patients with SHP the area under the curve for the first peak (mean 29.5%, range 20.6 to 40.4%) was significantly greater than that observed for patients with PHP (mean 15.6%, range 5.6 to 21.9%). This confirms previous studies showing accumulation of molecular forms of slightly lower molecular weight, presumably PTH (7-84), in patients with SHP and, to a lesser extent, in patients with PHP. The real necessity of assays that discriminate between these two molecular forms is debatable.

Skin secretion of Siphonops paulensis (Gymnophiona, Amphibia) forms voltage-dependent ionic channels in lipid membranes

The effect of the skin secretion of the amphibian Siphonops paulensis was investigated by monitoring the changes in conductance of an artificial planar lipid bilayer. Skin secretion was obtained by exposure of the animals to ether-saturated air, and then rinsing the animals with distilled water. Artificial lipid bilayers were obtained by spreading a solution of azolectin over an aperture of a Delrin cup inserted into a cut-away polyvinyl chloride block. In 9 of 12 experiments, the addition of the skin secretion to lipid bilayers displayed voltage-dependent channels with average unitary conductance of 258 ± 41.67 pS, rather than nonspecific changes in bilayer conductance. These channels were not sensitive to 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid or tetraethylammonium ion, but the experimental protocol used does not permit us to specify their characteristics.

Clinical classification of tetanus patients

The authors propose a clinical classification to monitor the evolution of tetanus patients, ranging from grade I to IV according to severity. It was applied on admission and repeated on alternate days up to the 10th day to patients aged > or = 12 years admitted to the State University Hospital, Recife, Brazil. Patients were also classified upon admission according to three prognostic indicators to determine if the proposed classification is in agreement with the traditionally used indicators. Upon admission, the distribution of the 64 patients among the different levels of the proposed classification was similar for the groups of better and worse prognosis according to the three indicators (P > 0.05), most of the patients belonging to grades I and II of the proposed classification. In the later reclassifications, severe forms of tetanus (grades III and IV) were more frequent in the categories of worse prognosis and these differences were statistically significant. There was a reduction in the proportion of mild forms (grades I and II) of tetanus with time for the categories of worse prognostic indicators (chi-square for trend: P = 0.00006, 0.03, and 0.00000) whereas no such trend was observed for the categories of better prognosis (grades I and II). This serially used classification reflected the prognosis of the traditional indicators and permitted the comparison of the dynamics of the disease in different groups. Thus, it becomes a useful tool for monitoring patients by determining clinical category changes with time, and for assessing responses to different therapeutic measures.

Leptin levels in different forms of Chagas' disease

Leptin is produced primarily by adipocytes. Although originally associated with the central regulation of satiety and energy metabolism, increasing evidence indicates that leptin may be an important mediator in cardiovascular pathophysiology. The aim of the present study was to investigate plasma leptin levels in patient with Chagas' heart disease and their relation to different forms of the disease. We studied 52 chagasic patients and 30 controls matched for age and body mass index. All subjects underwent anthropometric, leptin and N-terminal pro-brain natriuretic peptide (NT-proBNP) measurements and were evaluated by echocardiography, 12-lead electrocardiogram (ECG), and chest X-ray. All patients had fasting blood samples taken between 8:00 and 9:00 am. Chagasic patients were divided into 3 groups: group I (indeterminate form, IF group) consisted of 24 subjects with 2 positive serologic reactions for Chagas' disease and no cardiac involvement as evaluated by chest X-rays, ECG and two-dimensional echocardiography; group II (showing ECG abnormalities and normal left ventricular systolic function, ECG group) consisted of 14 patients; group III consisted of 14 patients with congestive heart failure (CHF group) and left ventricular dysfunction. Serum leptin levels were significantly lower (P < 0.001) in the CHF group (1.4 ± 0.8 ng/mL) when compared to the IF group (5.3 ± 5.3 ng/mL), ECG group (9.7 ± 10.7 ng/mL), and control group (8.1 ± 7.8 ng/mL). NT-proBNP levels were significantly higher (P < 0.001) in the CHF group (831.8 ± 800.1 pg/mL) when compared to the IF group (53.2 ± 33.3 pg/mL), ECG group (83.3 ± 57.4 pg/mL), and control group (32 ± 22.7 pg/mL). Patients with Chagas' disease and an advanced stage of CHF have high levels of NT-ProBNP andlow plasma levels of leptin. One or more leptin-suppressing mechanisms may operate in chagasic patients.

Influence of levels and forms of selenium associated with levels of vitamins C and E on the performance, yield and composition of tilapia fillet

The objective of this study was to evaluate the influence of vitamin C (VC), vitamin E (VE), and two selenium sources on the performance, yield, and composition of Nile tilapia fillet. The experiment was conducted in a completely randomized design consisting of six treatments with the addition of 100, 200, and 400 mg kg-1 VC and VE and 0.1, 0.2, and 0.4 of two sources of selenium. Each treatment had five replicates of 30 fish each. The diet with 200 mg kg-1 VC and VE + 0.2 mg.k-1 organic selenium resulted in weight gain, length gain, and feed conversion ratio similar to that of the treatment with 400 mg kg-1 VC and VE + 0.4 mg kg-1 organic or inorganic selenium. The addition of organic selenium to the diet improved the weight gain and feed conversion ratio in comparison with the addition of inorganic selenium. The diet with 0.2 mg kg-1 organic selenium showed glutathione peroxidase level equal to the diet with 0.4 mg kg-1 of inorganic selenium. Carcass and fillet yields were not affected by treatments; however, there was treatment effect on the fillet's chemical composition.