Hsp70 overexpression sequesters AIF and reduces neonatal hypoxic/ischemic brain injury.

Apoptosis is implicated in neonatal hypoxic/ischemic (H/I) brain injury among various forms of cell death. Here we investigate whether overexpression of heat shock protein (Hsp) 70, an antiapoptotic protein, protects the neonatal brain from H/I injury and the pathways involved in the protection. Postnatal day 7 (P7) transgenic mice overexpressing rat Hsp70 (Tg) and their wild-type littermates (Wt) underwent unilateral common carotid artery ligation followed by 30 mins exposure to 8% O(2). Significant neuroprotection was observed in Tg versus Wt mice on both P12 and P21, correlating with a high level of constitutive but not inducible Hsp70 in the Tg. More prominent injury was observed in Wt and Tg mice on P21, suggesting its continuous evolution after P12. Western blot analysis showed that translocation of cytochrome c, but not the second mitochondria-derived activator of caspase (Smac)/DIABLO and apoptosis-inducing factor (AIF), from mitochondria into cytosol was significantly reduced in Tg 24 h after H/I compared with Wt mice. Coimmunoprecipitation detected more Hsp70 bound to AIF in Tg than Wt mice 24 h after H/I, inversely correlating with the amount of nuclear, but not cytosolic, AIF translocation. Our results suggest that interaction between Hsp70 and AIF might have reduced downstream events leading to cell death, including the reduction of nuclear AIF translocation in the neonatal brains of Hsp70 Tg mice after H/I.     

Rat forebrain neurogenesis and striatal neuron replacement after focal stroke

The persistence of neurogenesis in the forebrain subventricular zone (SVZ) of adult mammals suggests that the mature brain maintains the potential for neuronal replacement after injury. We examined whether focal ischemic injury in adult rat would increase SVZ neurogenesis and direct migration and neuronal differentiation of endogenous precursors in damaged regions. Focal stroke was induced in adult rats by 90-minute right middle cerebral artery occlusion (tMCAO). Cell proliferation and neurogenesis were assessed with bromodeoxyuridine (BrdU) labeling and immunostaining for cell type-specific markers. Brains examined 10-21 days after stroke showed markedly increased SVZ neurogenesis and chains of neuroblasts extending from the SVZ to the peri-infarct striatum. Many BrdU-labeled cells persisted in the striatum and cortex adjacent to infarcts, but at 35 days after tMCAO only BrdU-labeled cells in the neostriatum expressed neuronal markers. Newly generated cells in the injured neostriatum expressed markers of medium spiny neurons, which characterize most neostriatal neurons lost after tMCAO. These findings indicate that focal ischemic injury increases SVZ neurogenesis and directs neuroblast migration to sites of damage. Moreover, neuroblasts in the injured neostriatum appear to differentiate into a region-appropriate phenotype, which suggests that the mature brain is capable of replacing some neurons lost after ischemic injury.     

Conversion of stable renal allograft recipients to a bioequivalent cyclosporine formulation

BACKGROUND: Gengraf capsule, an AB-rated generic cyclosporine for Neoral, has been shown to be bioequivalent in previous studies. The purpose of this pharmacokinetic study performed in stable renal transplant recipients was to evaluate interchangeability of Gengraf and Neoral. METHODS: Using an open-label, three-period design, 50 renal transplant recipients taking stable doses of Neoral completed a multicenter study. Subjects continued their Neoral regimen during period I (days 1-14). Subjects then switched from Neoral on a milligram-for-milligram basis to Gengraf during period II (days 15-28), followed by conversion to the same milligram-for-milligram dosing regimen of Neoral during period III (days 29-35). Twelve-hour pharmacokinetic evaluations (maximum observed blood concentration [C(max) ], concentration before dosing [C(trough) ], time to maximum observed concentration [T(max) ], and area under the blood concentration-vs.-time curve [AUC]) occurred on days 1, 14, 15, 28, and 29. Additional predose samples (C (trough)) were evaluated on days 7, 21, and 35. Laboratory and safety parameters were also evaluated. RESULTS: The pharmacokinetics of Gengraf (C(max), T(max), C(trough), and AUC) were indistinguishable from the Neoral values in stable renal allograft recipients. The bioequivalent capsules were interchangeable with respect to C(max), C(trough), and AUC at steady state and also on conversion from one capsule formulation to the other. The 90% confidence intervals (CI) for the Gengraf versus Neoral comparison at steady state (day 28 vs. day 14) were 0.95 to 1.03 for AUC and 0.92 to 1.04 for C(max). Trough concentrations remained consistent throughout the study, with no need for dosage adjustment in any of the subjects. Gengraf is well tolerated, with an excellent safety profile, comparable to the safety profile of Neoral. CONCLUSIONS The pharmacokinetics of Gengraf are equivalent and indistinguishable from those of Neoral. Gengraf is well tolerated and interchangeable with Neoral in stable renal transplant recipients.     

Sex differences in the effect of body-composition variables on bone mass in healthy children and adolescents

BACKGROUND: Prophylactic interventions against osteoporosis require a determination of the factors that influence the accumulation of bone mass during growth. OBJECTIVE: The objective was to determine the independent sex-specific contribution of lean mass and fat mass to bone mineral content (BMC), after adjustment for anthropometric variables and lifestyle factors, in healthy children and adolescents. DESIGN: Healthy schoolchildren (184 boys and 179 girls) aged 10-17 y (x+/- SD: 13.0 +/- 2.1 y) participated in this cross-sectional study. Total and regional (lumbar spine, femoral neck, and distal one-third of the radius) BMC and body composition were measured by dual-energy X-ray absorptiometry. RESULTS: A significant effect of anthropometric variables and lifestyle factors on BMC was observed at all skeletal sites. Lean mass and fat mass showed robust correlations with BMC, even after adjustment for anthropometric variables and lifestyle factors. Lean mass contributed to 6-12% of the variance in BMC in boys and to 4-10% in girls. Fat mass accounted for 0.1-2% of BMC variance in boys and to 0.1-6.5% in girls. CONCLUSIONS: Both lean mass and fat mass are consistent predictors of BMC at multiple skeletal sites in healthy children and adolescents. The contribution of lean mass to BMC variance was larger in boys than in girls. In both sexes, the highest contribution of lean mass to BMC was observed at the femoral neck.     

Prevalence of hepatitis B infection markers in Lebanese children: the need for an expanded programme on immunization

This multi-centre, cross-sectional study was designed to reveal the present status of hepatitis B infection markers among Lebanese children, and provide recommendations regarding childhood immunization policies. A total of 841 children, aged between 6 months and 6.5 years, were enrolled from Lebanon's five districts. Their sera were tested for hepatitis B surface antigen and hepatitis B core IgG. The overall prevalence of hepatitis B virus infection markers was 0.8% with increasing age-specific rates from 0% at 6 months to 1.3 % at > 5 years. There was no statistically significant association between the presence of hepatitis B markers and family characteristics or risk factors for infection. The highest prevalence rates were among children from Beirut suburbs (2.9 %) and South Lebanon (1.6%). The risk of horizontal transmission of hepatitis B to uninfected children increased substantially after the age of 2 years. An expanded programme on immunization that integrates hepatitisB vaccine during the first year of life is needed.     

In Vitro Assessment of the Hemocompatible Properties of Polymers

Summary: Based on the available data in the literature and personal results, a system of in vitro tests, consisting of two levels of selection, was proposed for assessment of hemocompatible properties of polymers. The first level consists of rapid methods for preliminary assessment (clotting time, hemolytic activity of the sample, and number of adhesive platelets). The second level involves methods for final selection and consists of measurement of the kinetics of the initial stages of adsorption of albumin, the degree of activation of the complement system, the intensity of radical oxidation of the lipid fraction of the blood, and the degree of platelet spreading. Comparative experimental assessment of these methods was conducted with a number of polymers and revealed the confidence limits for the application of each test and suggested quantitative criteria for selection.     

When bone becomes marble: Head and neck manifestations of osteopetrosis

Osteopetrosis is a genetically determined bone disease resulting from malfunction of osteoclastic activity, leading to excessive deposition of immature bone. This may result in complete agenesis of the paranasal sinuses, oral complications and multiple cranial neuropathies. The case of a 12-year-old boy with osteopetrosis is presented.     

Assessment of active transport of HIV protease inhibitors in various cell lines and the in vitro blood--brain barrier

OBJECTIVE: To investigate the involvement of P-glycoprotein (Pgp) and the multidrug resistance-associated protein (MRP) on the active transport of the HIV protease inhibitors amprenavir, ritonavir and indinavir. METHODS: The transport behaviour of ritonavir, indinavir and amprenavir in the presence and absence of Pgp modulators and probenecid was investigated in an in vitro blood--brain barrier (BBB) co-culture model and in monolayers of LLC-PK1, LLC-PK1:MDR1, LLC-PK1:MRP1 and Caco-2 cells. RESULTS: All three HIV protease inhibitors showed polarized transport in the BBB model, LLC-PK1:MDR1 and Caco-2 cell line. The Pgp modulators SDZ-PSC 833, verapamil and LY 335979 inhibited polarized transport, although their potency was dependent on both the cell model and the HIV protease inhibitor used. Ritonavir and indinavir also showed polarized transport in the LLC-PK1 and LLC-PK1:MRP1 cell line, which could be inhibited by probenecid. HIV protease inhibitors were not able to inhibit competitively polarized transport of other HIV protease inhibitors in the LLC-PK1:MDR1 cell line. CONCLUSIONS: Amprenavir, ritonavir and indinavir are mainly actively transported by Pgp, while MRP also plays a role in the transport of ritonavir and indinavir. This indicates that inhibition of Pgp could be useful therapeutically to increase HIV protease inhibitor concentrations in the brain and in other tissues and cells expressing Pgp. The HIV protease inhibitors were not able to inhibit Pgp-mediated efflux when given simultaneously, suggesting that simultaneous administration of these drugs will not increase the concentration of antiretroviral drugs in the brain.     

Effects of prolonged and exclusive breastfeeding on child height, weight, adiposity, and blood pressure at age 6.5 y: evidence from a large randomized trial

BACKGROUND: The evidence that breastfeeding protects against obesity and a variety of chronic diseases comes almost entirely from observational studies, which have a potential for bias due to confounding, selection bias, and selective publication. OBJECTIVE: We assessed whether an intervention designed to promote exclusive and prolonged breastfeeding affects children's height, weight, adiposity, and blood pressure at age 6.5 y. DESIGN: The Promotion of Breastfeeding Intervention Trial (PROBIT) is a cluster-randomized trial of a breastfeeding promotion intervention based on the WHO/UNICEF Baby-Friendly Hospital Initiative. A total of 17,046 healthy breastfed infants were enrolled from 31 Belarussian maternity hospitals and their affiliated clinics; of those infants, 13,889 (81.5%) were followed up at 6.5 y with duplicate measurements of anthropometric variables and blood pressure. Analysis was based on intention to treat, with statistical adjustment for clustering within hospitals or clinics to permit inferences at the individual level. RESULTS: The experimental intervention led to a much greater prevalence of exclusive breastfeeding at 3 mo in the experimental than in the control group (43.3% and 6.4%, respectively; P < 0.001) and a higher prevalence of any breastfeeding throughout infancy. No significant intervention effects were observed on height, body mass index, waist or hip circumference, triceps or subscapular skinfold thickness, or systolic or diastolic blood pressure. CONCLUSIONS: The breastfeeding promotion intervention resulted in substantial increases in the duration and exclusivity of breastfeeding, yet it did not reduce the measures of adiposity, increase stature, or reduce blood pressure at age 6.5 y in the experimental group. Previously reported beneficial effects on these outcomes may be the result of uncontrolled confounding and selection bias.