Relationship between intracranial pressure and antifungal agents levels in the CSF of patients with cryptococcal meningitis.

The purpose of this study was to evaluate the influence of intracranial hypertension in the cerebrospinal fluid (CSF) levels of amphotericin B and fluconazole levels of patients with cryptococcal meningitis. CSF samples and intracranial pressure were obtained by means of routine punctures performed at days 1, 7, and 14 of therapy, respectively. Amphotericin B and fluconazole CSF levels were measured by HPLC method as previously described. The minimum inhibitory concentration for amphotericin B, fluconazole, 5΄flucytosine, and voriconazole of each Cryptococcus isolate was performed according to CLSI. The predominant Cryptococcus species found was C. neoformans, and the major underlying condition was AIDS. Only one CSF sample had a detectable level for amphotericin B during the 14 days of therapy. Fluconazole CSF levels progressively increased from day 1 to day 14 of therapy for most cases. Fluconazole levels in the CSF were above the minimum inhibitory concentrations (MICs) for Cryptococcus during the initial 14 days of antifungal therapy. Variations of intracranial pressure did not affect amphotericin B and fluconazole levels in the CSF. The generalized estimating correlation (GEE) and Spearman correlation test (SCT) showed no significant correlation between the amphotericin B or fluconazole concentrations in the CSF and intracranial pressure (P = .953 and P = .093, respectively for GEE test and P = .477 and P = .847, respectively, for SCT). Combination therapy of amphotericin B with fluconazole was effective in 60% of the patients considering CSF cultures were negative in 9 of 15 patients after 14 days of therapy. Further studies are necessary to evaluate the role of intracranial hypertension on the therapeutic efficacy of different antifungal agents in patients with cryptococcal meningitis.

The beneficial effects of a water-based aerobic exercise session on the blood lipids of women with dyslipidemia are independent of their training status.

OBJECTIVES: To evaluate the acute effects of a session of water-based aerobic exercise on the blood lipid levels of women with dyslipidemia and to compare these results according to their training status. METHOD: Fourteen premenopausal women with dyslipidemia, aged 40-50 years, participated in two water-based aerobic exercise sessions, the first when they were generally sedentary and the second after they were trained with a water-based aerobic training program for 12 weeks. Both experimental sessions were performed using the same protocol, lasted 45 min, and incorporated an interval method, alternating 3 min at a rating of perceived exertion (RPE) of 13 and 2 min at an RPE of 9. Total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), and lipoprotein lipase enzyme (LPL) were obtained through venous blood collection before and immediately after each session. A generalized estimating equation method and Bonferroni tests were conducted (with time and training status as factors) for statistical analyses. RESULTS: At enrollment, the mean age of the participants was 46.57 years (95% confidence interval [CI] 44.81-48.34). The statistical analyses showed a significant time effect for all variables (TC: p=0.008; TG: p=0.012; HDL: p<0.001; LPL: p<0.001) except for LDL (p=0.307). However, the training status effect was not significant for any variable (TC: p=0.527; TG: p=0.899; HDL: p=0.938; LDL: p=0.522; LPL: p=0.737). These results indicate that the TC and TG levels reduced and the HDL and LPL concentrations increased from pre- to post-session in similar magnitudes in both sedentary and trained women. CONCLUSIONS: A single water-based aerobic exercise session is sufficient and effective to beneficially modify the lipid profile of women with dyslipidemia, regardless of their training status.

Effect of resistive exercise on muscle damage in water and on land.

The purpose of this study was to compare the effects of resistance exercise in water and on land on blood levels of creatine kinase (CK), a known indirect marker of muscle damage. Nine men (age: 23 +/- 1.58 years; weight: 79.37 +/- 11.15 kg; height: 176.33 +/- 4.09 cm), who had not practiced resistance training for at least 6 months before this experiment, performed 3 sets of maximum elbow flexion and extension exercises in water and on land, separated by 2-minute periods of rest. A 10-repetition maximum test was used to load control on land with free weights, and in water the exercise was performed at maximum velocity with aquatic resistance equipment. The duration of the exercise in water was the same as that performed on land, to reproduce the same metabolic route. Plasma CK activity was determined using a commercially available kit (Labtest Enzymatic-UV) before exercise, after exercise, and at 24 and 48 hours after exercise. Significant increases in CK were found at 48 hours postexercise on land (preexercise land: 160.75 +/- 96.05 UxL; 48 hours: 326.87 +/- 240.84 UxL), and significant differences were found between land and water (preexercise water: 147.75 +/- 46.48 UxL; 48 hours: 121.75 +/- 30.86 UxL) for this measure. However, no significant differences were found in water. In conclusion, the water environment influenced the absence of significant muscle damage. This type of exercise protocol may be appropriate for situations in which limited muscle tissue damage is desired.

Water-Based Aerobic Training Successfully Improves Lipid Profile of Dyslipidemic Women: A Randomized Controlled Trial.

PURPOSE: This study aimed to investigate the effects of water-based aerobic training on the lipid profile and lipoprotein lipase (LPL) levels in premenopausal women with dyslipidemia. METHOD: Forty women were randomly assigned to: aquatic training (WA; n = 20) or a control group (CG; n = 20). The WA group underwent 12 weeks of water-based interval aerobic training twice a week at intensities ranging from 9 to 15 on the Borg Scale of Perceived Exertion. Total cholesterol (TC), triglycerides (TG), high- (HDL) and low- (LDL) density lipoprotein, TC/HDL ratio, LPL levels, and peak oxygen consumption (VO2peak) were evaluated before and after 12 weeks in both groups. RESULTS: The WA group elicited decreases in TC (9%; effect size [ES] = 0.69; 95% CI [0.05, 1.33]), LDL (16%; ES = 0.78; 95% CI [0.13, 1.42]), and the TC/HDL ratio (17%; ES = 1.13; 95% CI [0.46, 1.79]), as well as increases in VO2peak (10%; ES = 0.64; 95% CI [0.002, 1.27]) and HDL (10%; ES = 0.28; 95% CI [-0.35. 0.90]), without significant changes in TG (ES = 0.16; 95% CI [-0.46, 1.79]) and LPL (ES = 0.36; 95% CI [-0.27, 0.98]) levels. In the CG, no statistically significant changes in any of these variables were found (TC, ES = 0.19, 95% CI [-0.43, 0.82]; LDL, ES = 0.22, 95% CI [-0.40, 0.85]; HDL, ES = 0.05, 95% CI [-0.57, 0.67]; TG, ES = 0.09, 95% CI [-0.53, 0.71]; TC/HDL ratio, ES = 0.20, 95% CI [-0.42, 0.82]; LPL, ES = 0.02, 95% CI [-0.60, 0.64]; VO2peak, ES = 0.20, 95% CI [-0.42, 0.82]). CONCLUSION: Water-based interval aerobic training positively affected the lipid profile in premenopausal dyslipidemic women.

The beneficial effects of a water-based aerobic exercise session on the blood lipids of women with dyslipidemia are independent of their training status

OBJECTIVES: To evaluate the acute effects of a session of water-based aerobic exercise on the blood lipid levels of women with dyslipidemia and to compare these results according to their training status. METHOD: Fourteen premenopausal women with dyslipidemia, aged 40-50 years, participated in two water-based aerobic exercise sessions, the first when they were generally sedentary and the second after they were trained with a water-based aerobic training program for 12 weeks. Both experimental sessions were performed using the same protocol, lasted 45 min, and incorporated an interval method, alternating 3 min at a rating of perceived exertion (RPE) of 13 and 2 min at an RPE of 9. Total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), and lipoprotein lipase enzyme (LPL) were obtained through venous blood collection before and immediately after each session. A generalized estimating equation method and Bonferroni tests were conducted (with time and training status as factors) for statistical analyses. RESULTS: At enrollment, the mean age of the participants was 46.57 years (95% confidence interval [CI] 44.81−48.34). The statistical analyses showed a significant time effect for all variables (TC: p=0.008; TG: p=0.012; HDL: p<0.001; LPL: p<0.001) except for LDL (p=0.307). However, the training status effect was not significant for any variable (TC: p=0.527; TG: p=0.899; HDL: p=0.938; LDL: p=0.522; LPL: p=0.737). These results indicate that the TC and TG levels reduced and the HDL and LPL concentrations increased from pre- to post-session in similar magnitudes in both sedentary and trained women. CONCLUSIONS: A single water-based aerobic exercise session is sufficient and effective to beneficially modify the lipid profile of women with dyslipidemia, regardless of their training status.

Kinetic studies on the inhibition of creatine kinase activity by branched-chain alpha-amino acids in the brain cortex of rats.

Maple syrup urine disease (MSUD) is a metabolic disorder biochemically characterized by the accumulation of branched-chain alpha-amino acids (BCAA) and their branched-chain alpha-keto acids (BCKA) in blood and tissues. Neurological dysfunction is usually present in the patients, but the mechanisms of brain damage in this disease are far from be understood. The main objective of this study was to investigate the mechanisms by which BCAA inhibit creatine kinase activity, a key enzyme of energy homeostasis, in the brain cortex of 21-day-old Wistar rats. For the kinetic studies, Lineweaver-Burk and a modification of the Chevillard et al. plots were used to characterize the mechanisms of enzyme inhibition. The results indicated that BCAA inhibit creatine kinase by competition with the substrates phosphocreatine and ADP at the active site. Considering the crucial role creatine kinase plays in energy homeostasis in brain, if these effects also occur in the brain of MSUD patients, it is possible that inhibition of this enzyme activity may contribute to the brain damage found in this disease. In this case, it is possible that creatine supplementation to the diet might benefit MSUD patients.

Effect of leucine administration on creatine kinase activity in rat brain.

Maple syrup urine disease (MSUD) is a metabolic disorder biochemically characterized by the accumulation of branched-chain amino acids (BCAA) and their branched-chain keto acids (BCKA) in blood and tissues. Neurological dysfunction is usually present in the patients, but the pathophysiology of brain damage is still obscure. Considering that brain energy metabolism is possibly altered in MSUD, the main objective of this study was to determine creatine kinase activity in the brain of rats subjected to acute and chronic administration of leucine. Chronic hyperleucinemia was induced by subcutaneous administrations of 4.8 micromol leucine/g body weight, twice a day, from the 6th to the 21st postnatal day. For acute hyperleucinemia, 21-day-old rats received three administrations of the amino acid at 3 h interval. Twelve hours after the chronic treatment or 1 h after the acute one, rats were killed and creatine kinase activity measured. The results indicated that acute or chronic administration of leucine altered creatine kinase activity in the brain of leucine-treated rats. Considering the crucial role creatine kinase plays in energy homeostasis in brain, if these effects also occur in the brain of MSUD patients, it is possible that alteration of this enzyme activity may contribute to the brain damage found in this disease.

Creatine kinase activity from rat brain is inhibited by branched-chain amino acids in vitro.

Maple syrup urine disease (MSUD) is an inherited metabolic disorder biochemically characterized by the accumulation of branched-chain amino acids (BCAAs) and their branched-chain keto acids (BCKAs) in blood and other tissues. Neurological dysfunction is usually present in the affected patients, but the mechanisms of brain damage in this disease are not fully understood. Considering that brain energy metabolism seems to be altered in MSUD, the main objective of this study was to investigate the in vitro effect of BCAAs and BCKAs on creatine kinase activity, a key enzyme of energy homeostasis, in brain cortex of young rats. BCAAs, but not their BCKAs, significantly inhibited creatine kinase activity at concentrations similar to those found in the plasma of MSUD patients (0.5-5 mM). Considering the crucial role creatine kinase plays in energy homeostasis in brain, if this effect also occurs in the brain of MSUD patients, it is possible that inhibition of this enzyme activity may contribute to the brain damage found in this disease.

Osso liofilizado bovino não-desmineralizado com células-tronco mesenquimais para engenharia tecidual: estudo experimental em sítio heterotópico / Bovine non-demineralized lyophilized bone with mesenchymal stem cells for tissue engineering: experimental study in heterotopic site

Objetivo: Avaliar o uso de células da medula óssea, com potencial osteogênico, agregadas a estrutura tridimensional de osso liofilizado bovino não-desmineralizado para engenharia tecidual óssea. Método: Os animais doadores de células da medula óssea, assim como os animais receptores dos construtos ósseos, foram camundongos de linhagem isogênica C57Bl/6. Utilizou-se modelo experimental heterotópico, com a implantação de construtos de osso liofilizado bovino não-desmineralizado (OL) no plano subcutâneo no dorso dos animais. Foram organizados 4 grupos de comparação (n=10 animais em cada grupo): 1) OL isoladamente (grupo controle); 2) OL + células mononucleares da medula (CMM); 3) OL + células-tronco mesenquimais (CTM); 4) OL + células-tronco mesenquimais diferenciadas em meio osteoindutor (CTMdif). A aferição foi realizada após 5 semanas, com avaliação histológica e determinação da atividade de fosfatase alcalina. Resultados: A avaliação histológica não mostrou diferença entre os grupos de comparação, com a observação em todas as amostras de tecido conjuntivo fibroso rico em neovasos estendendo-se por entre as trabéculas ósseas, sem osteoblastos ou osteócitos viáveis e sem neoformação óssea. Os resultados da atividade de fosfatase alcalina também não mostraram diferença entre os grupos de comparação, com análise de variância entre os grupos mostrando p=0,867. Conclusões: Os resultados mostraram que, no modelo estudado e com os métodos utilizados, a adição de células da medula óssea com potencial osteogênico sobre estrutura de osso liofilizado bovino não-desmineralizado não agregou propriedades osteogênicas ao material. Este estudo não confirmou a perspectiva inicial de utilizá-lo como estrutura tridimensional e carreadora celular na engenharia tecidual óssea, sendo necessários estudos subseqüentes que o avaliem em outros modelos experimentais, e que explorem separadamente cada etapa metodológica que possa influir no sucesso da engenharia tecidual óssea.

Objective: To evaluate the use of bone marrow cells with osteogenic potential seeded on bovine nondemineralized lyophilized bone scaffolds for bone tissue engineering. Method: Bone marrow cells donors, as well as the receptors of the bone constructs were C57BI/6 isogenic line mice. A heterotopic experimental model was used, with implantation of the constructs into subcutaneous pouches on the backs of the animals. Four comparison groups were set (n=10 animals each group): 1) LB alone (control group); 2) LB + marrow mononuclear cells (MMC); 3) LB + mesenchymal stem cells (MST); 4) LB + mesenchymal stem cells differentiated in osteoinductive medium (MSTdif). The constructs were harvested 5 weeks after implantation for histological analysis and alkaline phosphatase activity test. Results: The histological analysis did not show differences among the comparison groups. In all samples fibrous connective tissue rich in neovessels was observed extending through bone trabeculae, without viable osteoblasts or osteocytes and without new bone formation. Likewise, results of the alkaline phosphatase activity have not shown any difference among comparison groups, with the analysis of variance between groups showing p value=0.867. Conclusions: In this experimental model and with the methods used, the addition of bone marrow cells with osteogenic potential to a bovine non-demineralized lyophilized bone structure did not add osteogenic properties to the material. The initial perspective of using it as a scaffold for bone tissue engineering could not be confirmed, and further studies are required to assess it in other experimental models, and to explore separately each methodological step that might influence the success of bone tissue engineering.

Avaliaçäo da maturidade pulmonar fetal / Evaluation of fetal lung maturity

A avaliacao pulmonar laboratorial da maturidade pulmonar fetal através dos furfactantes é bastante útil na obstetrícia quando necessitamos interromper gestaçÝo pré-termo. Os autores discutem diversos testes descritos na literatura...