Fucosidosis and anesthesia

Fucosidosis is a rare, autosomal recessive lysosomal storage disorder caused by a severe deficiency of a-L-fucosidase. Patients usually have some problems with glycoprotein storage in the brain and other organs, and some structural abnormalities that need special consideration in anesthesia. It has 2 types, the early onset or infantile, and the juvenile. Here we present an 8-year old girl with deformities in the maxillofacial region, with big tongue, small and retracted chin, saddle nose, and short neck that could not be extended, causing difficult intubation, and congenital cardiac problems requiring a special anesthetic strategy.     

An image analysis approach for automatic malignancy determination of prostate pathological images

BACKGROUND: Determining malignancy of prostate pathological samples is important for treatment planning of prostate cancer. Traditionally, this is performed by expert pathologists who evaluate the structure of prostate glands in the biopsy samples. However, this is a subjective task due to inter- and intra-observer differences among pathologists. Also, it is time-consuming and difficult to some extent. Therefore, automatic determination of malignancy of prostate pathological samples is of interest. METHODS: A texture-based technique is first used to segment the prostate glands in the image. Features related to size and shape of these glands are then extracted and combined to generate an index, which is proportional to malignancy of cancer. A linear classifier is employed to classify the specimens into benign (low potential for malignancy) and malignant. RESULTS: The leave-one-out technique is employed to evaluate the method using two datasets. The first has 91 images with similar magnifications and illuminations while the second has 199 images with different magnifications and illuminations. In the experiments, accuracies of about 98 and 95% have been achieved for these two datasets, respectively. CONCLUSIONS: An image analysis approach is employed to evaluate prostate pathological images. Experimental results show that the proposed method can successfully classify the prostate biopsy samples into benign and malignant. They also show that the proposed method is robust to variations in magnification and illumination.     

Bone differentiation of marrow-derived mesenchymal stem cells using beta-tricalcium phosphate-alginate-gelatin hybrid scaffolds

The aim of the present study was to establish a 3D culture system for bone differentiation of mesenchymal stem cells (MSCs), using a new hybrid sponge. To manufacture the scaffold, a composite of beta-tricalcium phosphate-alginate-gelatin was prepared and cast as pellets of 1 cm diameter. The sponge was then fabricated by drying in freeze-dryer for 12 h. The porosity, mean pore size, compressive modulus and strength of the composite sponge fabricated in this study were 89.7%, 325.3 microm, 1.82 and 0.196 MPa, respectively. To establish a 3D culture system, the rat bone marrow-derived MSCs were suspended in 500 microl diluted collagen gel, loaded into the porous sponge and provided with medium with or without osteogenic supplements for 3 weeks. The day after loading, the cells appeared in the scaffold's internal spaces, where later some of them from either culture survived by anchoring on the surfaces. At the end of cultivation period, individually adhered cells from both cultures were observed to be replaced by cell aggregates, in which mineralized matrix was detected by alizarin red staining. Furthermore, RT-PCR analysis indicated that the bone-specific gene osteocalcin was expressed in cultures in both the presence and absence of the osteogenic supplements. Taken together, it seems that the studied scaffolds are cell-compatible and, more importantly, possess some osteo-inductive properties.     

Risk stratification and utilisation of thrombo-embolism prophylaxis in a medical-surgical ICU: a hospital-based study

To assess the risk for venous thrombo-embolism (VTE) and the utilisation of VTE prophylaxis in an open medical-surgical intensive care unit (ICU), patients were admitted and enrolled to the ICU at a multispecialty, tertiary care teaching hospital in Mumbai, for an observational study during a 3-month period. Risk factors for VTE and methods of VTE prophylaxis used were noted. Risk stratification was done, and the consultants attending the ICUs filled a questionnaire about VTE awareness, preferences and usage of VTE prophylaxis methods and reasons for not using thromboprophylaxis. Of 580 admissions, 488 were included in the study. As per the risk stratification, thromboprophylaxis was indicated in 466 (95%). The study group had an average VTE risk of 5.74, with a mean risk of 6.48 in medical patients and 5.0 in surgical patients. The most common risk factors in medical patients were bed rest >72 hours (56%), age >60 years (49.6%) and age 40-60 years (38.4%). Among surgical patients, the most common risk factors were major surgery (80.25%), central venous access (59.24%) and age 40-60 years (46.6%). Overall, VTE prophylaxis was used in 229 patients (47%), with 127 (55%) medical and 102 (45%) surgical patients. The most common methods of VTE prophylaxis used were elastic stockings (24.2%), low molecular weight heparin (15%) and low molecular weight heparin and stockings both (9.6%). Fear of bleeding was the commonest reason cited for the underutilisation of VTE prophylaxis. Almost half of all high-risk patients admitted to the ICU didn't receive any thromboprophylaxis. Consistent practice patterns of ICUs, and continuing medical education programmes addressing VTE prophylaxis will help improve the usage of VTE prophylaxis.     

Mechanisms of H2O2-induced oxidative stress in endothelial cells exposed to physiologic shear stress

Hydrogen peroxide (H2O2) is produced by inflammatory and vascular cells and induces oxidative stress, which may contribute to vascular disease and endothelial cell dysfunction. In smooth muscle cells, H2O2 induces production of O2 by activating NADPH oxidase. However, the mechanisms whereby H2O2 induces oxidative stress in endothelial cells are not well understood, although O2 may play a role. Recent studies have documented increased O2 in endothelial cells exposed to H2O2 via uncoupled nitric oxide synthase (NOS) and NADPH oxidase under static conditions. To assess responses to H2O2 in porcine aortic endothelial cells (PAEC) under shearing conditions, a constant flow rate of 24. 4 ml/min was applied to produce physiologically relevant shear stress (8. 2 dynes/cm). Here we demonstrate that treatment with 100 muM H2O2 increases intracellular O2 levels in PAEC. In addition, we demonstrate that l-NAME, an inhibitor of NOS, and apocynin, an inhibitor of NADPH oxidase, reduced O2 levels in PAEC treated with H2O2 under physiologic shear suggesting that both NOS and NADPH oxidase contribute to H2O2-induced O2 in PAEC. Co-inhibition of NOS and NADPH oxidase also reduced intracellular O2 levels under shear. We conclude that H2O2-induced oxidative stress in endothelial cells exhibits increased intracellular O2 levels through NOS and NADPH oxidase under shear. The inhibition of NOS and NADPH with H2O2 exposure is nonlinear, suggesting some interdependent or compensating system within endothelial cells. These findings suggest a complex interaction between H2O2 and oxidant-generating enzymes that may contribute to endothelial dysfunction in cardiovascular diseases.     

Prevalence and functionality of paucimorphic and private MC4R mutations in a large, unselected European British population, scanned by meltMADGE

Identification of unknown mutations has remained laborious, expensive, and only viable for studies of selected cases. Population-based "reference ranges" of rarer sequence diversity are not available. However, the research and diagnostic interpretation of sequence variants depends on such information. Additionally, this is the only way to determine prevalence of severe, moderate, and silent mutations and is also relevant to the development of screening programs. We previously described a system, meltMADGE, suitable for mutation scanning at the population level. Here we describe its application to a population-based study of MC4R (melanocortin 4 receptor) mutations, which are associated with obesity. We developed nine assays representing MC4R and examined a population sample of 1,100 subjects. Two "paucimorphisms" were identified (c.307G>A/p.Val103Ile in 27 subjects and c.-178A>C in 22 subjects). Neither exhibited any anthropometric effects, whereas there would have been >90% power to detect a body mass index (BMI) effect of 0.5 kg/m(2) at P=0.01. Two "private" variants were also identified. c.335C>T/p.Thr112Met has been previously described and appears to be silent. A novel variant, c.260C>A/p.Ala87Asp, was observed in a subject with a BMI of 31.5 kg/m(2) (i.e., clinically obese) but not on direct assay of a further 3,525 subjects. This mutation was predicted to be deleterious and analysis using a cyclic AMP (cAMP) responsive luciferase reporter assay showed substantial loss of function of the mutant receptor. This population-based mutation scan of MC4R suggests that there is no severe MC4R mutation with high prevalence in the United Kingdom, but that obesity-causing MC4R mutation at 1 in 1,100 might represent one of the commonest autosomal dominant disorders in man.     

Regulation of T-type calcium channels by Rho-associated kinase

We investigated the regulation of T-type channels by Rho-associated kinase (ROCK). Activation of ROCK via the endogenous ligand lysophosphatidic acid (LPA) reversibly inhibited the peak current amplitudes of rat Ca(v)3.1 and Ca(v)3.3 channels without affecting the voltage dependence of activation or inactivation, whereas Ca(v)3.2 currents showed depolarizing shifts in these parameters. LPA-induced inhibition of Ca(v)3.1 was dependent on intracellular GTP, and was antagonized by treatment with ROCK and RhoA inhibitors, LPA receptor antagonists or GDPssS. Site-directed mutagenesis of the Ca(v)3.1 alpha1 subunit revealed that the ROCK-mediated effects involve two distinct phosphorylation consensus sites in the domain II-III linker. ROCK activation by LPA reduced native T-type currents in Y79 retinoblastoma and in lateral habenular neurons, and upregulated native Ca(v)3.2 current in dorsal root ganglion neurons. Our data suggest that ROCK is an important regulator of T-type calcium channels, with potentially far-reaching implications for multiple cell functions modulated by LPA.