Polymorphism of gamma-adducin gene in genetic hypertension and mapping of the gene to rat chromosome 1q55

Adducin (ADD) is a heterodimeric protein involved in cellular signal transduction. A mutation in the alpha subunit affects ion transport and blood pressure in primary hypertension of Milan rats (MHS) and humans. In rats this effect is modulated by another mutation in the beta subunit. The recently described gamma subunit is a new member of the ADD family that should take the place of beta subunit in cells and tissues expressing alpha but not beta-Add. A missense mutation (Q572K) has been found in the gamma subunit of the Milan rats. Nineteen normotensive and five hypertensive inbred rat strains were genotyped for the polymorphisms in alpha, beta and gamma-Add genes. A disequilibrium was evident in the distribution of MHS-like Add genotype, being more frequent between the hypertensive than the normotensive strains (Chi-Square = 13.03, p = 0.0003). In kidney, brain, spleen, liver and heart a cDNA differing from gamma subunit by an in-frame insertion of 96 nucleotides, was found by PCR amplification and confirmed by RNase protection analysis. The rat gamma-Add gene was localized to chromosome 1q55 by fluorescence in situ hybridization.     

The carboxy-terminal region of human lipoprotein lipase is necessary for its exit from the endoplasmic reticulum

Certain missense substitutions on the human lipase (hLPL) gene produce mutated proteins that are retained in different compartments along the secretory pathway. The purpose of the present study was to elucidate whether the C-terminal domain of the hLPL molecule could be important for secretion. We constructed by site-directed mutagenesis three carboxy-terminal mutated (F388-->Stop, K428-->Stop and K441-->Stop) hLPL cDNAs that were expressed in COS1 cells. Immunoblotting of cell extracts showed that all three constructs led to similar levels of protein. Both wild type (WT) hLPL and the truncated K441-->Stop hLPL were secreted to the extracellular medium, and presented a similar intracellular distribution pattern as shown by immunofluorescence. Neither F388-->Stop nor K428-->Stop hLPL protein was detected in cell medium. Immunofluorescence experiments showed that both truncated hLPL were retained within an intracellular compartment, which became larger. Double immunofluorescence analysis using antibodies against LPL and antiprotein disulfide isomerase as a marker showed that the truncated K428-->Stop hLPL was retained within the rough endoplasmic reticulum. This truncated protein was not found in other compartments in the secretory pathway, such as Golgi complex and lysosomes, indicating that it did not exit the endoplasmic reticulum. Further analysis of the C-terminal region of the LPL molecular model showed both that F388-->Stop and K428-->Stop hLPL truncated proteins are highly hydrophobic. As retention of secretory proteins in the rough endoplasmic reticulum is a quality control mechanism of the secretory pathway, we conclude that the C-terminal domain of hLPL is critical for correct intracellular processing of the newly synthesized protein.     

Electron microscopy of the lesions produced in the human dura mater by Quincke beveled and Whitacre needles

INTRODUCTION: Comparisons of Quincke needles and non traumatic "pencil point" needles in recent years have reported lower rates of post dural puncture headache using the later type. Our new understanding of the morphology of the human dura mater motivated us to study dural lesions caused by the Whitacre 25 G and Quincke 26 G needles, using scanning electron microscopy with the aim of determining whether there is an anatomic basis for the different outcomes. METHOD: The dura mater from three fresh cadavers of individuals aged 65, 70 and 72 years were punctured 40 times at an angle of 90 degrees each time. The Whitacre 25 G needle was used for 20 punctures and the Quincke 26 G needle was used for the other 20. Half the punctures were performed with the bevel in the parallel alignment and the other half with the bevel perpendicular to the spinal column. Fifteen min after causing the punctures, specimens were fixed in solutions of glutaraldehyde phosphate buffer and dehydrated in acetone. After critical point removal of the acetone, after the specimens were treated with carbon and metallized with gold. The lesions were examined externally and internally and expressed as the ratio of area of lesion to diameter of the needle that had caused them. RESULTS: Whitacre needle: each lesion consisted in the superimposition of multiple damaged layers that started to close individually. After 15 min the outermost layers were 90% closed and the innermost ones had closed entirely. Layers in the arachnoid surface of the dura mater had closed from 86 to 88%, while deeper layers in the thick part had closed 97 to 98%. Quincke needle: lesions were V-shaped or half-moon shaped, much like the opening formed by a can opener, on both the external and internal surfaces. Alignment of the bevel of the needle parallel to the spinal column did not lead to a different shape of puncture. After 15 min the lesions had closed 94 to 95% on the epidural surface and 95 to 96% on the arachnoid side, a difference attributable to the retraction of the arachnoid layers over the spinal column. CONCLUSION: Non traumatic beveled dural needles, termed "pencil point needles", only partially separate dural fibers, and lesions caused by these needles develop in a more complex way. The Quincke 26G needle produced a puncture that is morphologically different from that caused by the Whitacre 25G needle, although lesions produced by both types close more than 94% after 15 min. We believe the size of the lesion caused by these needles does not explain the difference in post dural puncture headache due to loss of spinal fluid.     

Integrating a collaboration script and group awareness to support group regulation and emotions towards collaborative problem solving

International Journal of Computer-Supported Collaborative Learning - The research landscape displays increasing awareness of the important role of self-regulation and emotions in the process of...     

The Chemokine Receptor CCR3 Is Potentially Involved in the Homing of Prostate Cancer Cells to Bone: Implication of Bone-Marrow Adipocytes

Bone metastasis remains the most frequent and the deadliest complication of prostate cancer (PCa). Mechanisms leading to the homing of tumor cells to bone remain poorly characterized. Role of chemokines in providing navigational cues to migrating cancer cells bearing specific receptors is well established. Bone is an adipocyte-rich organ since 50 to 70% of the adult bone marrow (BM) volume comprise bone marrow adipocytes (BM-Ads), which are likely to produce chemokines within the bone microenvironment. Using in vitro migration assays, we demonstrated that soluble factors released by human primary BM-Ads are able to support the directed migration of PCa cells in a CCR3-dependent manner. In addition, we showed that CCL7, a chemokine previously involved in the CCR3-dependent migration of PCa cells outside of the prostate gland, is released by human BM-Ads. These effects are amplified by obesity and ageing, two clinical conditions known to promote aggressive and metastatic PCa. In human tumors, we found an enrichment of CCR3 in bone metastasis vs. primary tumors at mRNA levels using Oncomine microarray database. In addition, immunohistochemistry experiments demonstrated overexpression of CCR3 in bone versus visceral metastases. These results underline the potential importance of BM-Ads in the bone metastatic process and imply a CCR3/CCL7 axis whose pharmacological interest needs to be evaluated.     

Characterization and thermal degradation of poly(d,l‐lactide‐co‐glycolide) composites with nanofillers

Chemical and thermal characterization of poly(d ,l ‐lactide‐co‐glycolide) (PLGA) composites filled with hydroxyapatite (HA) or carbon nanotubes (CNT) were evaluated by infrared spectroscopy, differential scanning calorimetry, thermogravimetry, and dynamic–mechanical–thermal analysis. The morphology and distribution of the nanoparticles were studied by transmission electron microscopy. The composites were prepared by solvent casting using 30% HA or 1, 3, and 5% of pristine and functionalized CNT as nanoparticles and PLGA 75:25 and PLGA 50:50 as copolymer matrix. The Coats–Redfern and E₂ function methodologies were used to calculate the reaction order and the activation energy (E_a) of the thermal degradation process. It was found that the addition of nanoparticles increased the glass transition temperature (T_g) of the composites. Also, higher degradation temperatures and E_a values were obtained for PLGA–HA composites and compared with the neat copolymer, and the opposite behavior was exhibited by PLGA–CNT composites. The thermal and mechanical properties were highly dependent on the morphology and dispersion of the filler. The functionalization process of CNT promoted, to some extent, a better distribution and dispersion of CNT into the matrix, and these composites exhibited a slight enhancement on storage modulus. On the other hand, PLGA–HA composites showed a good dispersion but no improvement on the storage modulus below T_g. POLYM. ENG. SCI., 2013. © 2012 Society of Plastics Engineers     

Comparative study of layer-by-layer deposition techniques for poly(sodium phosphate) and poly(allylamine hydrochloride)

An inorganic short chain polymer, poly(sodium phosphate), PSP, together with poly(allylamine hydrochloride), PAH, is used to fabricate layer-by-layer (LbL) films. The thickness, roughness, contact angle, and optical transmittance of these films are studied depending on three parameters: the precursor solution concentrations (10^-3 and 10^-4 M), the number of bilayers deposited (20, 40, 60, 80, and 100 bilayers), and the specific technique used for the LbL fabrication (dipping or spraying). In most cases of this experimental study, the roughness of the nanofilms increases with the number of bilayers. This contradicts the basic observations made in standard LbL assemblies where the roughness decreases for thicker coatings. In fact, a wide range of thickness and roughness was achieved by means of adjusting the three parameters mentioned above. For instance, a roughness of 1.23 or 205 nm root mean square was measured for 100 bilayer coatings. Contact angles close to 0 were observed. Moreover, high optical transmittance is also reported, above 90%, for 80 bilayer films fabricated with the 10^-4 M solutions. Therefore, these multilayer structures can be used to obtain transparent superhydrophilic surfaces.     

Ring Opening of Decalin Over Zeolite-Supported Iridium Catalysts

Selective ring opening of decalin was investigated over various metal-modified zeolites in order to shed light on the key parameters affecting the selectivity to ring-opening products at low extent of cracking reactions. Ir-modified zeolites H-Y and H-Beta having different concentration and strength of acid sites were studied in a batch reactor at 250–310 °C and 20–60 bar H₂ and their performance was compared with that of Pt-modified zeolites. The concentration of Brønsted acid sites, reaction temperature and hydrogen pressure were shown to be the crucial reaction parameters. To obtain high selectivity to formation of ring-opening products from decalin, mildly acidic Ir-modified Beta was the best choice when the reaction was carried out at temperature of 270 °C or lower and at H₂ pressure of 60 bar. Under these conditions the selectivity and yield of ring-opening products exceeded 65 and 60%, respectively.