Pioglitazone improves the phenotype and molecular defects of a targeted Pkd1 mutant

Mutations of either PKD1 or PKD2 are associated with autosomal dominant polycystic kidney disease (ADPKD). The molecular function of the gene product of PKD1, polycystin-1, in vitro has been elucidated recently, but the molecular pathological consequences of the loss of polycystin-1 in vivo have remained unclear. We have generated a mouse with a targeted deletion of exons 2-6 of Pkd1 to study the molecular defects in Pkd1 mutants. Homozygote embryos (Pkd1(-/-)) developed hydrops, cardiac conotruncal defects and renal cystogenesis. Total protein levels of beta-catenin in heart and kidney and c-MYC in heart were decreased in Pkd1(-/-) embryos. In the kidneys of Pkd1(-/-), the expression of E-cadherin and PECAM in basolateral membranes of renal tubules was attenuated, and tyrosine phosphorylation of epidermal growth factor receptor and Gab1 were constitutively enhanced when cystogenesis started on embryonic day (E) 15.5-16.5. Maternally administered pioglitazone, a thiazolidinedione compound, resolved these molecular defects of Pkd1(-/-). Treatment with pioglitazone improved survival of Pkd1(-/-) embryos and ameliorated the cardiac defects and the degree of renal cystogenesis. Long-term treatment with pioglitazone improved the endothelial function of adult Pkd1(+/-). These data indicated that molecular defects observed in Pkd1(-/-) embryos contributed to the pathogenesis of ADPKD and that thiazolidinediones had a compensatory effect on the pathway affected by the loss of polycystin-1. Pathways activated by thiazolidinediones may provide new therapeutic targets in ADPKD.     

Apathy and depression in cross-cultural survivors of traumatic brain injury

The disturbance of motivation and its relationship to depression continues to spark contradictory findings among European and North American populations. Could a cross-cultural study shed some light on the situation? This study aims to detect the prevalence of apathy and to test whether the Apathy Evaluation Scale (AES) can spot the presence or absence of depression in survivors of traumatic brain injury (TBI) in Oman. Eighty subjects who sustained a TBI were given an Arabic version of the AES and were also interviewed with the semistructured Composite International Diagnostic Interview (CIDI). The authors found that the incidence of apathy and depression among Omani people who sustained TBI is similar to that reported elsewhere. The AES has poor discriminatory power in identifying cases of depression. These findings emphasize the importance of developing assessment tools that are culturally sensitive in light of the rising incidence of TBI in developing countries such as Oman.     

Signaling complex formation of phospholipase Cbeta4 with metabotropic glutamate receptor type 1alpha and 1,4,5-trisphosphate receptor at the perisynapse and endoplasmic reticulum in the mouse brain

Upon activation of cell surface receptors coupled to the Gq subclass of G proteins, phospholipase C (PLC) beta hydrolyses membrane phospholipid to yield a pair of second messengers, inositol 1,4,5-trisphosphate (IP3) and 1,2-diacylglycerol. PLCbeta4 has been characterized as the isoform enriched in cerebellar Purkinje cells (PCs) and the retina and involved in motor and visual functions. Here we examined cellular and subcellular distributions of PLCbeta4 in adult mouse brains. Immunohistochemistry showed that high levels of PLCbeta4 were detected in the somatodendritic domain of neuronal populations expressing the metabotropic glutamate receptor (mGluR) type 1alpha, including olfactory periglomerular cells, neurons in the bed nucleus anterior commissure, thalamus, substantia nigra, inferior olive, and unipolar brush cells and PCs in the cerebellum. Low to moderate levels were detected in many other mGluR1alpha-positive neurons and in a few mGluR1alpha-negative neurons. In PCs, immunogold electron microscopy localized PLCbeta4 to the perisynapse, at which mGluR1alpha is concentrated, and to the smooth endoplasmic reticulum in dendrites and spines, an intracellular Ca2+ store gated by IP3 receptors. In the cerebellum, immunoblot demonstrated its concentrated distribution in the post-synaptic density and microsomal fractions, where mGluR1alpha and type 1 IP3 receptor were also greatly enriched. Furthermore, PLCbeta4 formed coimmunoprecipitable complexes with mGluR1alpha, type 1 IP3 receptor and Homer 1. These results suggest that PLCbeta4 is preferentially localized in the perisynapse and smooth endoplasmic reticulum as a component of the physically linked phosphoinositide signaling complex. This close molecular relationship might provide PLCbeta4 with a high-fidelity effector function to mediate various neuronal responses under physiological and pathophysiological conditions.     

Dorsal telencephalon-specific RA-GEF-1 knockout mice develop heterotopic cortical mass and commissural fiber defect

Neural migration defects lead to various types of human malformations of cortical development including subcortical band heterotopia, which shows formation of a secondary cortical plate beneath the primary cortex and is typically caused by mutation of the DCX ( doublecortin ) gene. Subcortical band heterotopia is usually associated with mental retardation and epilepsy. We previously discovered RA-GEF-1 as a guanine nucleotide exchange factor (GEF) for Rap1 small GTPase. Here we have analysed its in-vivo role in formation of the adult cerebral cortex by using telencephalon-specific RA-GEF-1 conditional knockout (cKO) mice, generated by mating RA-GEF-1 ^{flox / flox} mice with Emx1-cre knockin mice. RA-GEF-1 cKO mice showed severe defects in their brain structures including an ectopic cortical mass underlying a relatively normal cortex. The ectopic cortical mass lacked the normal six-layered lamination but preserved the subcortical connectivity as revealed by retrograde tracing. Further, RA-GEF-1 cKO mice exhibited a lower threshold for the induction of epileptic seizures. These phenotypes have a resemblance to those of human subcortical band heterotopia. In addition, the agenesis of anterior commissures, the dorsal hippocampus commissure, the corpus callosum and the enlargement of the lateral ventricles were observed in cKO mice. Our findings suggest a crucial function of RA-GEF-1 in neural migration.     

The Guinea pig as a preclinical model for demonstrating the efficacy and safety of statins

Statins, because of their excellent efficacy and manageable safety profile, represent a key component in the current armamentarium for the treatment of hypercholesterolemia. Nonetheless, myopathy remains a safety concern for this important drug class. Cerivastatin was withdrawn from the market for myotoxicity safety concerns. BMS-423526 [{(3R,5S)-7-[4-(4-fluorophenyl)-6,7-dihydro-2-(1-methylethyl)-5H-benzo[6,7]cyclohepta[1,2-b]pyridin-3-yl]-3,5-dihydroxy-heptenoic acid} sodium salt], similar to cerivastatin in potency and lipophilicity, was terminated in early clinical development due to an unacceptable myotoxicity profile. In this report, we describe the guinea pig as a model of statin-induced cholesterol lowering and myotoxicity and show that this model can distinguish statins with unacceptable myotoxicity profiles from statins with acceptable safety profiles. In our guinea pig model, both cerivastatin and BMS-423526 induced myotoxicity at doses near the ED(50) for total cholesterol (TC) lowering in plasma. In contrast, wide differences between myotoxic and TC-lowering doses were established for the currently marketed, more hydrophilic statins, pravastatin, rosuvastatin, and atorvastatin. This in vivo model compared favorably to an in vitro model, which used statin inhibition of cholesterol synthesis in rat hepatocytes and L6 myoblasts as surrogates of potential efficacy and toxicity, respectively. Our conclusion is that the guinea pig is a useful preclinical in vivo model for demonstrating whether a statin is likely to have an acceptable therapeutic safety margin.     

Association between body mass index and functional independence measure in patients with deconditioning

OBJECTIVE: To assess the association of body mass index (BMI) with functional independence measure (FIM) score in patients with deconditioning. We also examined whether the association was different for motor and cognitive subscales of the FIM instrument. DESIGN: A retrospective study of 1077 inpatients admitted to the general medicine service for deconditioning at an acute rehabilitation hospital. Patients were classified into underweight (BMI < 18.5), normal range (BMI = 18.5-24.9), overweight (BMI = 25.0-29.9), obese class I (BMI = 30.0-34.9), obese class II (BMI = 35.0-39.9), and obese class III (BMI > or = 40). RESULTS: Median gain in FIM scores from admission to discharge was highest in obese class I patients (27 points), followed by obese class II patients (26 points). The most gain in FIM scores was accounted for by the motor subscale. Adjusting for age, gender, and length of in-hospital stay, obese class I patients had a 5.8-point (95% confidence limits = 1.2, 7.0) higher gain in FIM score compared with patients with BMI in the normal range. CONCLUSIONS: In an acute rehabilitation setting, obese patients had higher gains in FIM scores as compared with normal-range-BMI patients. Most of the improvements in FIM scores were accounted for by the motor subscale, with little or no improvement on the cognitive scale.     

The effect of methylphenidate on the sleep-wake cycle of brain-injured patients undergoing rehabilitation

BACKGROUND AND PURPOSE: A number of neuro-stimulants are routinely used as part of post-acute care of hospitalized brain-injured patients. To our knowledge, the effect of these stimulants on the sleep-wake cycles of brain-injured patients undergoing rehabilitation has not been addressed. We examined the effect of one of the most commonly used neuro-stimulants, methylphenidate, on the sleep-wake behavior of brain-injured patients undergoing rehabilitation at a dedicated brain injury clinic. PATIENTS AND METHOD: For this study, records of patients admitted between January and December 1999 were scrutinized retrospectively for the data on observationally defined sleep-wake distribution. A total of 30 patients diagnosed with traumatic brain injury were identified as having been observed for a full 24h a day for at least 10 days. Some of these patients (n=17) were administered methylphenidate on clinical grounds. They served as the experimental group, while the unmedicated patients (n=13) served as controls. For the present analysis, the sleep-wake cycles were arbitrarily designated as nighttime and daytime, respectively. A cumulative sleep-wake quantity in a 24-h period was also observed. RESULT: The average number of hours of sleep during a 24-h period was not significantly different for the two cohorts. Similar trends emerged for the nighttime and daytime observations. On the whole, methylphenidate appears not to have unfavorable effects on sleep-wake cycles, presently defined as nighttime, daytime and 24-h, in the traumatic brain injury population. CONCLUSION: This study sought to gain better understanding of the effect of methylphenidate on daytime sleepiness and nighttime sleep, and the data suggest that administration of methylphenidate does not appear to have an adverse effect on sleep-wake quantity.     

Dynamic distribution of muscle-specific calpain in mice has a key role in physical-stress adaptation and is impaired in muscular dystrophy

Limb-girdle muscular dystrophy type 2A (LGMD2A) is a genetic disease that is caused by mutations in the calpain 3 gene (CAPN3), which encodes the skeletal muscle–specific calpain, calpain 3 (also known as p94). However, the precise mechanism by which p94 functions in the pathogenesis of this disease remains unclear. Here, using p94 knockin mice (termed herein p94KI mice) in which endogenous p94 was replaced with a proteolytically inactive but structurally intact p94:C129S mutant protein, we have demonstrated that stretch-dependent p94 distribution in sarcomeres plays a crucial role in the pathogenesis of LGMD2A. The p94KI mice developed a progressive muscular dystrophy, which was exacerbated by exercise. The exercise-induced muscle degeneration in p94KI mice was associated with an inefficient redistribution of p94:C129S in stretched sarcomeres. Furthermore, the p94KI mice showed impaired adaptation to physical stress, which was accompanied by compromised upregulation of muscle ankyrin-repeat protein-2 and hsp upon exercise. These findings indicate that the stretch-induced dynamic redistribution of p94 is dependent on its protease activity and essential to protect muscle from degeneration, particularly under conditions of physical stress. Furthermore, our data provide direct evidence that loss of p94 protease activity can result in LGMD2A and molecular insight into how this could occur.     

缺血预处理对大鼠四动脉结扎后氧化应激及脑水肿的保护作 …

观察缺血预处理对大鼠四动脉结扎法的全脑缺血／再灌注损伤的保护作用。预处理后,缺血３０ｍｉｎ,再灌注１０及３０ｍｉｎ。记录并比较ＥＥＧ、ＭＤＡ、脑含水量和脑梗塞范围。脑缺血期间脑电波变平,缺血预处理减少脑电波的变化,并且减少梗塞范围。再灌注１０和３０ｍｉｎ后,对梗塞抑制率分别为２０％和５４％。缺血预处理也可将ＭＤＡ和含水量恢复到正常水平。预处理消除氧化应激反应和脑水肿,比减少脑梗塞范围更有效。