The role of His-18 in amyloid formation by human islet amyloid polypeptide

The 37-residue islet amyloid polypeptide (IAPP) is the major protein component of the amyloid deposits found in type-II diabetes. IAPP is stored in a relatively low pH environment in the pancreatic secretory granules prior to its release to the extracellular environment. Human IAPP contains a single histidine at position 18. Aggregation of IAPP is considerably faster at a lower pH (4.0 +/- 0.3) than at high pH (8.8 +/- 0.3), as judged by turbidity and thioflavine-T fluorescence studies. The rate of aggregation at low pH increases drastically in the presence of salt. CD experiments show that the conversion of largely unstructured monomers to beta-sheet-rich structures is faster at high pH. TEM studies show that fibrils are formed at both pH values but are more prevalent at pH 8.8 (+/-0.3). Both the free N terminus of IAPP and His-18 will titrate over the pH range studied. An N-terminal acetylated fragment consisting of residues 8-37 of human IAPP was also studied to isolate contributions from the protonation of His-18. Previous studies have shown that this fragment forms protofibrils that are very similar to those formed by intact IAPP. The effects of varying the protonation state of His-18 in the 8-37 analogue indicate that the rate of aggregation and fibril formation is noticeably faster when His-18 is deprotonated, similar to the wild type. However, the pH-dependent effects are larger for full-length IAPP than for the disulfide-truncated, acetylated analogue. TEM studies indicate differences in the morphology of the deposits formed at high and low pH. These results are discussed in light of recent structural models of IAPP fibrils.     

A defect in the ionotropic glutamate receptor 6 gene (GRIK2) is associated with autosomal recessive mental retardation

Nonsyndromic mental retardation is one of the most important unresolved problems in genetic health care. Autosomal forms are far more common than X-linked forms, but, in contrast to the latter, they are still largely unexplored. Here, we report a complex mutation in the ionotropic glutamate receptor 6 gene (GRIK2, also called “GLUR6”) that cosegregates with moderate-to-severe nonsyndromic autosomal recessive mental retardation in a large, consanguineous Iranian family. The predicted gene product lacks the first ligand-binding domain, the adjacent transmembrane domain, and the putative pore loop, suggesting a complete loss of function of the GLU_K6 protein, which is supported by electrophysiological data. This finding provides the first proof that GLU_K6 is indispensable for higher brain functions in humans, and future studies of this and other ionotropic kainate receptors will shed more light on the pathophysiology of mental retardation.     

Gamma–Radiation-Assisted Synthesis of Luminescent ZnO/Ag Heterostructure Core–Shell Nanocomposites

There is increasing interest in tuning the physical properties of semiconductor nanostructures using metal nanoparticles. In this work, ZnO nanosphere covered with Ag nanoparticles were synthesized using gamma–radiation-assisted method. The amount of deposited Ag nanoparticles is controlled by changing irradiation dose in the range of 30–100 kGy in order to tune the semiconductor–metal interaction. The successful deposition of Ag on the ZnO nanoparticles is examined by analyzing the morphology, microstructure, optical, and magnetic properties of ZnO/Ag nanoparticles through field emission scanning electron (FESEM), microscopy X-ray diffraction spectra, UV-visible absorption, photoluminescence measurement, and vibrating sample magnetometer. FESEM and elemental mapping results confirmed that Ag nanoparticles have been concentrated at the surface of spherical ZnO particles. Moreover, formation of pure metallic Ag nanoparticles has been confirmed by XRD analysis. UV-visible absorption spectra of obtained ZnO/Ag showed two combined peaks, a weak peak at the shoulder around 360 nm corresponds to ZnO and a sharp absorption at 420 nm refers to spherical Ag nanoparticles. Obtained results from photoluminescence revealed that the near-band-edge emission and defect-related visible emission bands of ZnO could be enhanced dramatically at the same time by deposition of Ag nanoparticles, which was ascribed to localized surface plasmon–exciton coupling and surface plasmon scattering. Controlling the semiconductor and metal coupling effect is interesting because of its application in highly efficient optoelectronic devices and biosensor.     

Antimicrobial susceptibility profiles associated with bacterial meningitis among children: a referral hospital-based study in Iran

Bacterial meningitis continues to be associated with high morbidity and mortality rate worldwide, especially in the pediatric age group. This study was performed to identify the microbial etiologies of meningitis among 31 children, who were admitted in the Emergency Ward of a referral pediatric hospital in Iran. Culture identification showed that Streptococcus pneumoniae (12 subjects), Haemophilus influenzae (11 subjects) were the most common bacteria, followed by Escherichia coli (7 cases) and Neisseria meningitidis (only one case). Antibiotic susceptibility tests revealed that vancomycin had the best effect on S. pneumoniae in comparison with other antibiotics, whereas H. influenzae and E. coli were more susceptible to ceftriaxone, ceftazidime, and ceftizoxime than other antibiotics. In conclusion, despite the advances in antibiotic therapy and vaccine development, bacterial meningitis still is a health problem. S. pneumoniae, H. influenzae, and N. meningitidis are the main sources of bacterial meningitis, but other organisms such as E. coli should also be suspected, when a case is admitted to a referral pediatric hospital.     

Effects of cryopreservation with a newly-developed magnetic field programmed freezer on periodontal ligament cells and pulp tissues

The purpose of this study was to evaluate the effects of long-term cryopreservation on the isolated human periodontal ligament cells (PDL) and pulp tissues. In the first part of study, 10 freshly extracted teeth were selected and divided into two groups. In the cryopreserved group, the teeth were frozen for 5 years using a programmed freezer combined with a magnetic field, known as Cells Alive System “CAS”. As for the control group, freshly extracted teeth were used. In each group, extracted PDL tissues were cultured and gene expression and protein concentration of collagen type I, alkaline-phosphatase (ALP) and vascular endothelial growth factor (VEGF) was compared between the two groups. In the second part, pulp tissues were obtained from 10 mature and immature third molars which were freshly extracted or cryopreserved for three months. Expression of VEGF and nerve growth factor (NGF) mRNAs and the protein concentration in the supernatant were investigated. Results indicated that long-term cryopreservation with the use of CAS freezer cannot affect the growth rate and characteristics of PDL cells. There was no significant difference in VEGF expression and VEGF and NGF protein concentration of pulp cells derived from cryopreserved teeth with immature apex and control group with mature root formation. Finally, proper PDL regeneration and appropriate apexogenesis after transplanting magnetically cryopreserved immature tooth was clinically confirmed. These findings demonstrate that teeth banking with the use of magnetic field programmed freezer can be available for future autotransplantation as a treatment modality for replacing missing teeth.     

Effects of Zinc Oxide Nanoparticles on Performance,Egg Quality,Tissue Zinc Content,Bone Parameters,and Antioxidative Status in Laying Hens

The objective of this study was to observe the effects of dietary supplementation with zinc oxide nanoparticles (ZnO-NPs) on performance, egg quality, tissue Zn content, bone parameters, superoxide dismutase activity (SOD), and egg malondialdehyde (MDA) content in laying hens. A total of 288 laying hens at 64 weeks of age were randomly assigned to 4 treatments, 6 replicates, with 12 birds each. Experimental diets included the based diet (without Zn supplementation), and basal diet supplemented with 40, 80, and 120 mg Zn/kg from ZnO-NPs. Feed intake and egg mass were significantly higher in the 40 and 80 ZnO-NPs groups than the other groups. The birds in the 80 ZnO-NPs group had significantly higher egg product than the 120 ZnO-NPs and control groups. Egg shell thickness and shell strength significantly increased in the 40 and 80 ZnO-NPs groups. Moreover, Haugh unit significantly improved in the groups supplemented with ZnO-NPs, compared to the control group. Bone-breaking strength was significantly greater in the 80 ZnO-NPs group than the 120 ZnO-NPs and control groups. Also, ash weight was significantly greater in the 40 and 80 ZnO-NPs groups than the control group. There were significant differences among the groups in the Zn content in plasma, tibia, liver, pancreas, and egg. Relative to the control group, ZnO-NPs supplementation significantly increased the activities of SOD in the liver, pancreas, and plasma. The MDA content in egg was significantly reduced in the groups supplemented with ZnO-NPs. In conclusion, this study demonstrates that ZnO-NPs as dietary supplementation can improve the performance of laying hens, and levels of 40 to 80 ZnO-NPs are the optimal concentrations.

     

Aggregation of islet amyloid polypeptide: from physical chemistry to cell biology

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Highlights? Islet amyloid contributes to type-2 diabetes and to islet transplant failure. ? IAPP is natively unfolded, but is one of the most amyloidogenic proteins known. ? High resolution models of IAPP amyloid fibers are available. ? Anionic lipids and glycosaminoglycans catalyze IAPP amyloid formation in vitro. ? There are multiple mechanisms of IAPP induced β-cell toxicity.     

Autosomal recessive mental retardation: homozygosity mapping identifies 27 single linkage intervals, at least 14 novel loci and several mutation hotspots

Mental retardation (MR) has a worldwide prevalence of around 2% and is a frequent cause of severe disability. Significant excess of MR in the progeny of consanguineous matings as well as functional considerations suggest that autosomal recessive forms of MR (ARMR) must be relatively common. To shed more light on the causes of autosomal recessive MR (ARMR), we have set out in 2003 to perform systematic clinical studies and autozygosity mapping in large consanguineous Iranian families with non-syndromic ARMR (NS-ARMR). As previously reported (Najmabadi et al. in Hum Genet 121:43-48, 2007), this led us to the identification of 12 novel ARMR loci, 8 of which had a significant LOD score (OMIM: MRT5-12). In the meantime, we and others have found causative gene defects in two of these intervals. Moreover, as reported here, tripling the size of our cohort has enabled us to identify 27 additional unrelated families with NS-ARMR and single-linkage intervals; 14 of these define novel loci for non-syndromic ARMR. Altogether, 13 out of 39 single linkage intervals observed in our cohort were found to cluster at 6 different loci on chromosomes, i.e., 1p34, 4q27, 5p15, 9q34, 11p11-q13 and 19q13, respectively. Five of these clusters consist of two significantly overlapping linkage intervals, and on chr 1p34, three single linkage intervals coincide, including the previously described MRT12 locus. The probability for this distribution to be due to chance is only 1.14 × 10(-5), as shown by Monte Carlo simulation. Thus, in contrast to our previous conclusions, these novel data indicate that common molecular causes of NS-ARMR do exist, and in the Iranian population, the most frequent ones may well account for several percent of the patients. These findings will be instrumental in the identification of the underlying genes.