On-line extraction and determination of carbofuran in raw milk by direct HPLC injection on an ISRP column

A method has been developed for extraction and determination of carbofuran in milk. The method involved direct injection of raw milk on to a human serum albumin dimethyloctyl-silica gel (HSA-Cs) column and the use of 80:20 (v/v) 0.01 M phosphate buffer pH 5.5 - acetonitrile as mobile phase. UV spectrophotometric detection was performed at 220 nm. Identification was based on retention time. Quantification was performed by automatic peak-area determination and was calibrated by use of an external standard.

High volume peritoneal dialysis for acute renal failure

Background. Peritoneal dialysis (PD) is still widely used for acute renal failure (ARF) in developing countries despite concerns about its inadequacy. Continuous PD has been evaluated in ARF by analyzing the resolution of metabolic abnormality and normalization of plasma pH, bicarbonate, and potassium.Methodology: A prospective study was performed on 30 ARF patients who were assigned to high-dose continuous PD (Kt/V = 0.65 per session) via a flexible catheter (Tenckhoff) and automated PD with a cycler. Fluid removal, pH and metabolic control, protein Loss, and patient outcome were evaluated.Results: Patients received 236 continuous PD sessions; 76% were admitted to ICUs. APACHE II score was 32.2 +/- 8.65. BUN concentrations stabilized after 3 sessions, creatinine after 4, and bicarbonate and pH after 2. Fluid removal was 2.1 +/- 0.62 L/day. Creatinine and urea clearances were 15.8 +/- 4.16 and 17.3 +/- 5.01 mL/minute respectively. Normalized creatinine clearance and urea Kt/V values were 110.6 +/- 22.5 L/week/1.73 m(2) body surface area and 3.8 +/- 0.6 respectively. Solute reduction index was 41% +/- 6.5% per session. Serum albumin values remained stable in spite of considerable protein tosses (median 21.7 g/day, interquartile range 9.1 - 29.8 g/day). Regarding ARF outcome, 23% of patients presented renal function recovery, 13% remained on dialysis after 30 days of follow-up, and 57% died.Conclusion: High-dose continuous PD by flexible catheter and cycler was an effective treatment for ARF. It provided high solute removal, allowing appropriate metabolic and pH control, and adequate dialysis dose and fluid removal. Continuous PD can therefore be considered an alternative to other forms of renal replacement therapy in ARF.

Hydrolyzed alpha-lactalbumin as a source of protein to the exercising

This work describes the effect of feeding enzymatically hydrolyzed a-lactalbumin on blood sugar, albumin and fatty acids, muscular and hepatic glycogen of rats subjected to physical exercise. Three normoenergetic/normoproteic diets, containing either casein (C), alpha-lactalbumin (L) or alpha-lactalbumin hydrolyzate (H) were fed to thirty male Wistar rats for five weeks. During this period, half of the rats swam for 1 hr daily (T category) while the other half remained sedentary (S category). At the end of training, all rats were required to swim to exhaustion. The results showed that those rats of the T-category consuming diet H reached exhaustion with significantly higher concentrations of serum glucose ([H] 56.0 and [L] 32.3 mg/100ml), serum albumin ([H] 3.8 and [L] 2.1 mg/dl) and muscle glycogen ([H] 2.1 and [L] 0.6 mg/g), while no differences were observed between diets regarding the time of arrival to exhaustion. Results from diets C and L differed minimally. It was concluded that feeding the hydrolyzed protein may result in nutritional advantage to the exercising rat. (C) 1998 Elsevier B.V.

Sulfation of intrinsic glycoproteins of the rabbit vitreous

The experiments reported here were designed to characterize the intrinsic vitreous glycoproteins and to understand the process of their sulfation. Rabbits were injected intravitreally with S-35-sodium sulfate and killed at several time intervals after injection. In another series of experiments, rabbits were injected either with S-35-sodium sulfate, H-3-fucose or H-3-tyrosine, associated or not associated with tunicamycin administration. Vitreous from the control eyes was also digested with N-glycosidase.. Furthermore, ciliary bodies, the putative source of the intrinsic vitreous glycoproteins, were incubated with S-35-sodium sulfate in the presence or absence of the protein synthesis inhibitor cycloheximide, and the culture media recovered for analysis. These and the vitreous samples of the other experiments were processed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography. Except for serum albumin, practically all polypeptide bands of the vitreous and culture media were labeled with radioactive sulfate and were shown to undergo renewal. The experiments using tunicamycin or enzyme treatment suggest that radioactive sulfate was incorporated not only into the carbohydrate side chains of the glycoproteins but also into the amino acid tyrosine of the polypeptide backbone of these glycoproteins. (C) 1998 Academic Press.

BIOSENSORS FOR GLUCOSE NEEDLE-SHAPED FOR IN-VIVO MONITORING

Different procedures for obtaining a needle biosensor for the determination of glucose to be inserted subcutaneously in vivo, have been compared. Platinum wires with a diameter of 75 mum, teflon-coated were inserted in hypodermic needles and fixed with a two-component epoxy resin. Using a dip-coating procedure, several layers were deposited on electrodes. The first coating was cellulose acetate, the second was immobilized glucose oxidase (GOD) mixed with bovine serum albumin (BSA) and glutaraldheyde, the third coating was a polyurethane coating obtained with commercially available products. A large number of electrodes have been tried and statistically evaluated but they seem to be affected by poor reproducibility evidenced by a large spreading in successive calibration curves. Then, the polyurethane coating has been replaced by a thin polycarbonate membrane salinized and fixed on the tip of the needle. Reproducible results were achieved and first results of in vivo measurements on rabbits are reported.

REGULATION OF BOVINE ENDOMETRIAL SECRETION OF PROSTAGLANDINS AND SYNTHESIS OF 2',5'-OLIGOADENYLATE SYNTHETASE BY INTERFERON-ALPHA MOLECULES

Experiments were performed to (1) verify the inhibitory effect of bovine trophoblast protein-1 (bTP-1) on uterine prostaglandin synthesis, (2) evaluate whether other interferon-alpha (IFN-alpha) molecules also inhibit prostaglandin secretion, and (3) test whether the enzyme 2',5'-oligoadenylate synthetase (2-5A synthetase) can be induced in endometrium by interferon-alpha. In experiment 1, all interferon molecules (bTP-1, oTP-1, bIFN-alpha and hIFN-alpha) equally inhibited secretion of PGF and PGE2 from endometrial explant cultures obtained at day 17 of the estrous cycle. In experiment 2, endometrial explants obtained from day 17 of the cycle were cultured with and without bovine serum albumin (BSA; 50-mu-g/ml) and bIFN-alpha (0, 0.84, 4.2, and 42 nM). Addition of BSA to the culture medium greatly enhanced the accumulation of PGF into the medium. The bIFN-alpha inhibited accumulation of PGF and PGE2 in both the presence or absence of BSA by 12 h. All three concentrations of bIFN-alpha were equally effective in inhibiting prostaglandin accumulation. Additionally, all concentrations of bIFN-alpha increased the amounts of 2-5A synthetase in endometrium. In conclusion, these results confirm the inhibitory effect of bTP-1 on PGF release from endometrium and demonstrate that bTP-1 can also inhibit PGE2 secretion. Furthermore, other interferon-alpha molecules, including bIFN-alpha, hIFN-alpha, and oTP-1, also reduced PGF and PGE2 secretion in culture. It is likely, therefore, that conceptus and other interferon-alpha molecules exert similar effects on endometrium in vitro and that the antiluteolytic effects of bIFN-alpha in vivo are mediated in part by changes in endometrial prostaglandin synthesis.

Protein deficiency during pregnancy and lactation impairs glucose-induced insulin secretion but increases the sensitivity to insulin in weaned rats

We studied glucose homeostasis in rat pups from darns fed on a normal-protein (170 g/kg) (NP) diet or a diet containing 60 g protein/kg (LP) during fetal life and the suckling period. At birth, total serum protein, serum albumin and serum insulin levels were similar in both groups. However, body weight and serum glucose levels in LP rats were lower than those in NP rats. At the end of the suckling period (28 d of age), total serum protein, serum albumin and serum insulin were significantly lower and the liver glycogen and serum free fatty acid levels were significantly higher in LP rats compared with NP rats. Although the fasting serum glucose level was similar in both groups, the area under the blood glucose concentration curve after a glucose load was higher for NP rats (859 (SEM 58) mmol/l per 120 min for NP rats v. 607 (SEM 52) mmol/l per 120 min for LP rats; P < 0.005). The mean post-glucose increase in insulin was higher for NP rats (30 (SEM 4.7) nmol/l per 120 min for NP rats v. 17 (SEM 3.9) nnol/l per 120 min for LP rats; P < 0.05). The glucose disappearance rate for NP rats(0.7 (SEM 0.1) %/min) was lower than that for LP rats (1.6 (SEM 0.2) %/min; P < 0.001). Insulin secretion from isolated islets (1 h incubation) in response to 16.7 mmol glucose/l was augmented 14-fold in NP rats but only 2.6-fold in LP rats compared with the respective basal secretion (2.8 mmol/l; P <0.001). These results indicate that in vivo as well as in vitro insulin secretion in pups from dams maintained on a LP diet is reduced. This defect may be counteracted by an increase in the sensitivity of target tissues to insulin.

Implicações nutricionais no tratamento e recuperação de pacientes adultos com diarréia crônica.

The nutritional assessment by 24 hour-dietary recall, anthropometry and blood-components measurements was undertaken in 23 adult patients, 17 males and 6 females suffering of chronic diarrhea from pancreatitis (30%), inflammatory bowel disease (22%), short intestine syndrome (9%) and unknown diarrhea (35%). The nutritional assessment was done at the entry and repeated at the discharge of the hospitalization that averaged 35 days, during which the patients received specific medical treatment along with obstipating diets. The hospitalization resulted in overall improvement of the patients either clinically by reducing their defecation rate or nutritionally by increasing their protein-energy intake and the values of anthropometry and blood components (albumin, free-tryptophan and lymphocytes). When the patients where divided into two groups based on their fecal-fat output one could note the better nutritional response of the group showing steatorrhea than the non-steatorrhea group, with the serum albumin and the arm-muscle circumference being discriminatory between groups. However even in the better recovered patients the indicative values of a satisfactory nutritional status were not accomplished. Thus, these data suggest that besides the overall nutritional improvement seen in the studied chronic diarrhea patients the full-nutrition recovering would demand either or both a longer hospitalization and/or an early-aggressive nutritional support.

The hemolytic uremic syndrome as a complication of adriamycin nephropathy

Rats treated with two injections of adriamycin (week 0 and week 12) developed glomerusclerosis and severe tubulointerstitial lesions as described in the literature. In addition, a number of glomerular alterations were present. These included capillary loop dilation, insudation of eosinophilic material, necrosis, duplication of the glomerular basement membrane, severe mesangiolysis with disruption of the mesangial matrix and segmental double- contours. The renal arterioles and interlobular arteries showed endothelial cell swelling. The subendothelial space was infiltrated by fibrinoid material and there was intensive fibrinoid necrosis of the wall of both arteries and arterioles extending into the glomerular tuft. These alterations were very similar to those observed in the hemolytic uremic syndrome. This observation suggests that the two injections of adriamycin, with a long interval in between them, might induce renal lesions similar to those observed in the hemolytic uremic syndrome.

Preparation of N2, N2,7-trimethylguanosine affinity columns

2,2,7-trimethylguanosine (TMG) binding proteins from human cells were purified through TMG-affinity columns. TMG synthesis was improved and the TMG obtained was shown to be similar to the TMG in the 5' cap of the UsnRNAs. The eluates obtained with TMG-affinity chromatographies were very different from those isolated with m7G-affinity columns, thus suggesting that specific TMG- binding proteins were obtained. The fraction may be enriched with factors associated with import and/or hypermethylation of UsnRNPs.

Historico da vacinacao antivariolica no Brasil

A historical chronology of smallpox vaccination in Brazil is presented, with emphasis on the State of Sao Paulo. We also present the scientific and philosophical concepts that influenced the regulation and practice of vaccination in Sao Paulo based on the historiographic bibliography, legislation about vaccination, and the debates in the state legislative body. Discovered by Jenner in 1796, the vaccine reached in Brazil in 1804 and was only used in the colonial capital, the city of Rio de Janeiro. In the 1890 decade, smallpox, side by side with yellow fever, typhoid fever and other pestilential diseases, was the major health problem in the State of Sao Paulo. There was also the fear that the vaccine might transmit syphilis, an Unfounded attitude since the product used in Sao Paulo (the 'animal vaccine') was elaborated from bovine serum. The immediate necessity to fight a highly lethal disease that threatened the State population and the coffee-growing business led to the abandonment of the fears and of the liberal principles in favor of the sanitary needs. The vaccine became compulsory in 1891 in the State of Sao Paulo and its application met no resistance on the part of the population, in contrast to the so-called 'Vaccine Revolt' that would occur in Rio de Janeiro in 1904.

Avaliações bioquímicas séricas e hepáticas do extrato de yacon (Polymnia sonchifolia) Poepp. & Endl.

Yacon is a medicinal plant, the tuberous roots of which have been thought to contain a large amount of fructan (oligosaccharides). Purpose - The aim of paper was to study the effect of aqueous extracts of yacon on biochemical parameters of clinical importance in rats. The animals (male, wistar, weighing approximately 300g) were divided in 3 groups: G1(n=8)= water control; G2(n=8)= aqueous extract of roots (0,17g/100g/day); G3(n=8)= aqueous extract of leaves (25mg/100g/day). The serum samples were obtained after 30 days, and the biochemical parameters were measured. The livers were removed and homogenized in 0,01M phosphate buffer pH 7,0 and then the supernatant fractions were used for enzyme assay. Significantly increased serum glucose was observed in G2 (206,72±91,27 mg/dL). The groups G2 and G3 rats had higher (p<0,05) urea concentration, while creatinin level decreased (p<0,05). The serum albumin concentration showed a tendency to remain in G2 (2,44±0,45 g/dL) and G3 (2,84±0,50 g/dL). Aqueous extracts administration markedly decreased (p<0,05) the activities of ALT in the liver and greatly increased these enzymatic activities in blood. The serum alkaline phosphatase activity increased (p<0,05) in rats receiving the aqueous extract of root (192,75±20,95 U/dL), while aqueous extract of leaves reduced it (129,57±19,93 U/dL). The results indicated that the extract of yacon promoted changes of the biochemical parameters.

Systemic hypertension in patients with glomerulonephritis

Although systemic hypertension is very common in patients with glomerulonephritis there is a dispute if this alteration is consequence of the glomerulonephritis per se or is a consequence of the renal failure secondary to the glomerular lesion. With the aim to analyze the factors associated with systemic hypertension, 196 patients with different forms of nephritis were studied. The systemic arterial pressure was measured by standard sphygmomanometer, renal function was evaluated by the determination of the serum creatinine concentration or creatinine clearance. The diagnosis of the type of glomerulonephritis was made on the basis of an examination of kidney biopsy specimens. The prevalence of arterial hypertension among patients with glomerulonephritis was 62.7%. The hypertensive patients were older (hypertensive = 30.6 ± 12.8; normotensive = 25.4 ± 1.6 years; P = 0.03). The prevalence of arterial hypertension was lower in patients with minimal glomerular lesion (12.5%), though their ages were also lower (18.1 ± 3.6 and 29.1 ± 1.03 years; P = 0.03). Arterial hypertension did not correlate with the serum levels of creatinine and albumin; creatinine clearance and 24-h proteinuria. In conclusion: In the patients with glomerulonephritis, the presence of arterial hypertension was associated with a higher mean age whereas the intensity of proteinuria, the level of renal function or the type of glomerulonephritis was not different between the two groups.

Action of anti-bothropic factor isolated from Didelphis marsupialis on renal effects of Bothrops erythromelas venom

Acute renal failure is the most common complication in the lethal cases caused by snakebites in Brazil. Among the Brazilian venom snakes, Bothrops erythromelas is responsible for the majority of accidents in Northeastern Brazil. Didelphis marsupialis serum could inhibit myonecrotic, hemorrhagic, edematogenic hyperalgesic and lethal effects of envenomation determined by ophidian bites. In the present study, we evaluated the action of the anti-bothropic factor isolated from D. marsupialis on the renal effects promoted by B. erythromelas venom without systemic interference. Isolated kidneys from Wistar rats were perfused with Krebs-Henseleit solution containing 6% bovine serum albumin. We analyzed renal perfusion pressure (PP), renal vascular resistance (RVR), glomerular filtration rate (GFR), urinary flow (UF), and the percentages of sodium and potassium tubular transport (%TNa +, %TK +). The B. erythromelas venom (10 μg mL -1) decreased the PP (ct=108.71±5.09 mmHg; BE=65.21±5.6 mmHg*) and RVR (ct=5.76±0.65 mmHg mL -1 g -1 min -1; BE=3.10±0.45 mmHg mL -1 g -1 min -1*) . On the other hand, the GFR decreased at 60 min (ct 60=0.76±0. 07 mL g -1 min -1; BE 60=0.42±0.12 mL g -1 min -1*) and increased at 120 min (ct 120=0.72±0.01 mL g -1 min -1; BE 120=1.24±0.26 mL g -1 min -1*). The UF increased significantly when compared with the control group (ct=0.14±0.01 mL g -1 min -1; BE=0.47±0.08 mL g -1 min -1*). The venom reduced the %TNa + (ct 90=79.18±0.88%; BE 90=58.35±4.86%*) and %TK + (ct 90=67.20±4.04%; BE 90=57. 32±5.26%*) The anti-bothropic factor from D. marsupialis (10 μg mL -1) incubated with B. erythromelas venom (10 μg mL -1) blocked the effects on PP, RVR, %TNa +, and %TK +, but was not able to reverse the effects in UF and GFR promoted by venom alone. However, the highest concentration of D. marsupialis serum (30 μg mL -1) reversed all the renal effects induced by the venom. In conclusion, B. erythromelas venom altered all the renal functional parameters evaluated and the anti-bothropic factor from D. marsupialis was able to inhibit the effects induced by the venom in isolated kidney. © 2005 Elsevier Ltd. All rights reserved.

Efeitos de diferentes sistemas de cultivo in vitro sobre o crescimento de folículos pré-antrais isolados de ovários de fetos bovinos

The objective of this study is to use different in vitro culture systems of preantral follicles from Nelore breed bovine fetuses in the last gestation quarter. The evaluation of treatments considered the time of growth of isolated follicles. Preantral follicles were mechanically isolated and submitted to the individual culture, for 9 days, in media no supplemented or supplemented with fetal calf serum (FCS), bovine serum albumin (BSA) or synthetic defined supplement substitute of serum KnockoutSR (KNO). We have also evaluated the effects of collagen gel or fetal calf fibroblast monolayer as substratum for in vitro cultures. The increase on the follicular diameter was followed in the first day (0 h), at the 72 h, 144 h and 216 h. Considering cultures of isolated follicles, the results have shown that the association between media supplemented with FCS and collagen gel was significantly more efficient on the increase of the follicular diameter than other treatments. It is not still established a system of appropriate cultivation that sustains the differentiation and multiplication of the granular cells and that maintains the contact of the same ones with the oocyte to provide molecules and factors that supply the metabolic demand. We also understand that our results also represent another promising step on the search for the ultimate system of in vitro culture of preantral follicles from bovines.