Intra-renal and subcellular distribution of the human chloride channel, CLC-5, reveals a pathophysiological basis for Dent's disease

Dent's disease, which is a renal tubular disorder characterized by low molecular weight proteinuria, hypercalciuria and nephrolithiasis, is associated with inactivating mutations of the X-linked chloride channel, CLC-5. However, the manner in which a functional loss of CLC-5 leads to such diverse renal abnormalities remains to be defined. In order to elucidate this, we performed studies to determine the segmental expression of CLC-5 in the human kidney and to define its intracellular distribution. We raised and characterized antisera against human CLC-5, and identified by immunoblotting an 83 kDa band corresponding to CLC-5 in human kidney cortex and medulla. Immunohistochemistry revealed CLC-5 expression in the epithelial cells lining the proximal tubules and the thick ascending limbs of Henle's loop, and in intercalated cells of the collecting ducts. Studies of subcellular human kidney fractions established that CLC-5 distribution was associated best with that of Rab4, which is a marker of recycling early endosomes. In addition, confocal microscopy studies using the proximal tubular cell model of opossum kidney cells, which endogenously expressed CLC-5, revealed that CLC-5 co-localized with the albumin-containing endocytic vesicles that form part of the receptor-mediated endocytic pathway. Thus, CLC-5 is expressed at multiple sites in the human nephron and is likely to have a role in the receptor-mediated endocytic pathway. Furthermore, the functional loss of CLC-5 in the proximal tubules and the thick ascending limbs provides an explanation for the occurrences of low molecular weight proteinuria and hypercalciuria, respectively. These results help to elucidate further the patho-physiological basis of the renal tubular defects of Dent's disease.     

Thermo‐economic‐environmental multiobjective optimization of a gas turbine power plant with preheater using evolutionary algorithm

In this study, the gas turbine power plant with preheater is modeled and the simulation results are compared with one of the gas turbine power plants in Iran namely Yazd Gas Turbine. Moreover, multiobjective optimization has been performed to find the best design variables. The design parameters of the present study are selected as: air compressor pressure ratio (r_AC), compressor isentropic efficiency (η_AC), gas turbine isentropic efficiency (η_GT), combustion chamber inlet temperature (T₃) and gas turbine inlet temperature. In the optimization approach, the exergetic, economic and environmental aspects have been considered. In multiobjective optimization, the three objective functions, including the gas turbine exergy efficiency, total cost rate of the system production including cost rate of environmental impact and CO₂ emission, have been considered. The thermoenvironomic objective function is minimized while power plant exergy efficiency is maximized using a genetic algorithm. To have a good insight into this study, a sensitivity analysis of the results to the interest rate as well as fuel cost has been performed. In addition, the results showed that at the lower exergetic efficiency in which the weight of thermoenvironomic objective is higher, the sensitivity of the optimal solutions to the fuel cost is much higher than the location of Pareto Frontier with the lower weight of thermoenvironomic objective. Copyright © 2010 John Wiley & Sons, Ltd.     

Declining sperm counts in the United States? A critical review

PURPOSE: Recent reports suggest declining sperm counts in the United States. These reports did not include all available data and did not account for geographic variations noted in prior studies. We examined all available data on U.S. sperm counts and evaluated whether geographic variations account for the decline suggested. MATERIALS AND METHODS: We reviewed all 29 U.S. studies from 1938 to 1996 reporting manually counted semen analyses of 9,612 fertile or presumably fertile men. We determined mean sperm concentrations by geographic location with weighted analysis of variance, and assessed any changes with time by linear regression analysis. RESULTS: Mean sperm concentrations from New York were significantly higher than from all other U.S. cities (98.6 versus 71.6 x 10(6) sperm per cc, respectively, p = 0.006). There has been no statistically significant change with time for mean sperm concentrations reported from New York (p = 0.49) or from U.S. cities other than New York (p = 0.62). Analysis without separating by location revealed a decline (p = 0.047). CONCLUSIONS: Sperm concentrations are highest in New York compared to other U.S. cities. When accounting for this geographic difference and examining all available data, there appears to be no significant change in sperm counts in the U.S. during the last 60 years. Further studies addressing the causes of geographic variations are needed.     

KinMutBase, a database of human disease-causing protein kinase mutations

KinMutBase (http://www.uta.fi/laitokset/imt/KinMut Base.html) is a registry of mutations in human protein kinases related to disorders. Kinases are essential cellular signalling molecules, in which mutations can lead into diseases including, e.g., immunodeficiencies, cancers and endocrine disorders. The first release of KinMutBase contains information for nine protein tyrosine kinases. There are altogether 170 entries representing 273 families and 403 patients. Mutations appear both in conserved hallmark residues of the kinases as well as in non-homologous sites. The KinMutBase WWW pages provide plenty of information, namely mutation statistics and display, clickable sequences with mutations, restriction enzyme patterns and online submission.     

Exact solution for stability analysis of moderately thick functionally graded sector plates on elastic foundation

In this article, an exact analytical solution for buckling analysis of moderately thick functionally graded (FG) sector plates resting on Winkler elastic foundation is presented. The equilibrium equations are derived according to the first order shear deformation plate theory. Because of the coupling between the bending and stretching equilibrium equations of FG plates, these plates have deflection under in-plane loads lower than the critical buckling load acting on the mid-plane. The conditions under which FG plates remain flat in the pre-buckling configuration are investigated and the stability equations are obtained based on the flat plate assumption in the pre-buckling state. The stability equations are simplified into decoupled equations and solved analytically for plates having simply supported boundary condition on the straight edges. The critical buckling load is obtained and the effects of geometrical parameters and power law index on the stability of functionally graded sector plates are studded. The results for the critical buckling load of moderately thick functionally graded sector plates resting on elastic foundation are reported for the first time.     

Heat Transfer Coefficient for a Packed Bed of Shredded Materials at Low Peclet Numbers

The application of the Nusselt number (Nu) correlation of a packed bed of spherical particles overestimates the rate of heat transfer in the packed beds of shredded materials. This is due to the dependence of flow distribution on the particle geometry. This discrepancy is even more pronounced due to a channeling effect at low Peclet numbers (Pe). In this study, the heat transfer coefficient for a packed bed of shredded materials was derived at low Peclet numbers. In deriving the correlation, both numerical simulation and experimental work were employed. The experimental work was carried out in a packed bed of combustible materials for the flow rates of 15 × 10^{? 6} to 50 × 10^{? 6} m ³/s and a temperature range of 30 to 60°C. Using the results of experimental work along with numerical modeling and applying a power law formulation, the best-fit Nusselt number for the packed bed of shredded materials for Pe < 25 was derived to be Nu = 0.015 0.11 + Pe ^0.73 . The Nusselt number calculated from this correlation is in good agreement with the experimental data.     

TP53 Targeted Deep Sequencing of Cell-Free DNA in Esophageal Squamous Cell Carcinoma Using Low-Quality Serum: Concordance with Tumor Mutation

Circulating cell-free DNA (cfDNA) is emerging as a potential tumor biomarker. CfDNA-based biomarkers may be applicable in tumors without an available non-invasive screening method among at-risk populations. Esophageal squamous cell carcinoma (ESCC) and residents of the Asian cancer belt are examples of those malignancies and populations. Previous epidemiological studies using cfDNA have pointed to the need for high volumes of good quality plasma (i.e., >1 mL plasma with 0 or 1 cycles of freeze-thaw) rather than archival serum, which is often the main available source of cfDNA in retrospective studies. Here, we have investigated the concordance of TP53 mutations in tumor tissue and cfDNA extracted from archival serum left-over from 42 cases and 39 matched controls (age, gender, residence) in a high-risk area of Northern Iran (Golestan). Deep sequencing of TP53 coding regions was complemented with a specialized variant caller (Needlestack). Overall, 23% to 31% of mutations were concordantly detected in tumor and serum cfDNA (based on two false discovery rate thresholds). Concordance was positively correlated with high cfDNA concentration, smoking history (p-value = 0.02) and mutations with a high potential of neoantigen formation (OR; 95%CI = 1.9 (1.11–3.29)), suggesting that tumor DNA release in the bloodstream might reflect the effects of immune and inflammatory context on tumor cell turnover. We identified TP53 mutations in five controls, one of whom was subsequently diagnosed with ESCC. Overall, the results showed that cfDNA mutations can be reliably identified by deep sequencing of archival serum, with a rate of success comparable to plasma. Nonetheless, 70% non-identifiable mutations among cancer patients and 12% mutation detection in controls are the main challenges in applying cfDNA to detect tumor-related variants when blindly targeting whole coding regions of the TP53 gene in ESCC.     

Stable association of PYK2 and p130(Cas) in osteoclasts and their co-localization in the sealing zone

Bone resorption is initiated by osteoclast attachment to the mineralized matrix, cytoskeletal reorganization, cellular polarization, and the formation of the sealing zone. The present study examines the interaction between PYK2 and p130(Cas) (Crk-associated substrate), suggested to be part of the signaling pathway initiated by osteoclast adhesion. Using murine osteoclast-like cells (OCLs) and their mononuclear precursors (pOCs), generated in a co-culture of bone marrow and osteoblastic MB1.8 cells, we show that: 1) p130(Cas) is tyrosine-phosphorylated upon adhesion of pOCs to vitronectin or ligation of beta3 integrins; 2) p130(Cas) colocalizes with PYK2 and the cytoskeletal proteins F-actin, vinculin, and paxillin in the podosomal-rich ring-like structures of OCLs plated on glass and in the sealing zone in actively resorbing OCLs on bone; 3) p130(Cas) and PYK2 form a stable complex in pOCs, independent of tyrosine phosphorylation of either molecule, and this complex is present in Src (-/-) OCLs, in which neither protein is phosphorylated or associated with the osteoclast adhesion structure; 4) the association of p130(Cas) and PYK2 is mediated by the SH3 domain of p130(Cas) and the C-terminal domain of PYK2. These findings suggest that p130(Cas) and its association with PYK2 may play an important role in the adhesion-dependent signaling that leads to cytoskeletal reorganization and formation of the sealing zone during osteoclast activation.