Structural and molecular changes in the aging choroid: implications for age-related macular degeneration.

Age-related macular degeneration (AMD) is a devastating disease-causing vision loss in millions of people around the world. In advanced stages of disease, death of photoreceptor cells, retinal pigment epithelial cells, and choroidal endothelial cells (CECs) are common. Loss of endothelial cells of the choriocapillaris is one of the earliest detectable events in AMD, and, because the outer retina relies on the choriocapillaris for metabolic support, this loss may be the trigger for progression to more advanced stages. Here we highlight evidence for loss of CECs, including changes to vascular density within the choriocapillaris, altered abundance of CEC markers, and changes to overall thickness of the choroid. Furthermore, we review the key components and functions of the choroid, as well as Bruch's membrane, both of which are vital for healthy vision. We discuss changes to the structure and molecular composition of these tissues, many of which develop with age and may contribute to AMD pathogenesis. For example, a crucial event that occurs in the aging choriocapillaris is accumulation of the membrane attack complex, which may result in complement-mediated CEC lysis, and may be a primary cause for AMD-associated choriocapillaris degeneration. The actions of elevated monomeric C-reactive protein in the choriocapillaris in at-risk individuals may also contribute to the inflammatory environment in the choroid and promote disease progression. Finally, we discuss the progress that has been made in the development of AMD therapies, with a focus on cell replacement.

When is facial diplegia regarded as a variant of Guillain-Barré syndrome?

A variant of Guillain-Barré syndrome (GBS) with predominant manifestation of facial diplegia (FD) has been described recently. This study aimed to characterize and determine the incidence of this FD-predominant GBS variant. The clinical and serological information of 900 consecutive patients were reviewed. In total, eight patients were identified between January 2007 and December 2010 as having FD accompanied by some features of GBS. These features were subjective sensory symptoms such as distal paresthesia (7/8, 88%), albumin-cytological (A/C) dissociation (7/8, 88%), antecedent infection (6/8, 75%), and minor nerve conduction study (NCS) abnormalities (5/7, 71%). One patient presented with the typical NCS feature of demyelinating neuropathy. Only two patients exhibited areflexia (2/8, 25%). None of the patients possessed any anti-ganglioside antibodies; however, the serum of two patients was positive for anti-mycoplasma antibody (2/6, 33%). FD variant of GBS occurred in less than 1% of our dataset. FD can be a regional variant of GBS when it is accompanied by supporting features, such as subjective tingling, A/C dissociation, and minor NCS abnormalities.

Expression of heat shock protein 90 in the kidneys of diabetic db/db mice.

OBJECTIVE: To identify novel genes regulated in diabetic nephropathy. MATERIALS AND METHODS: Total RNA from the renal cortex of db/+ and db/db mice was isolated and DNA microarrays specific for diabetes signaling pathways were used for expression profiling. Expression of mRNA and protein was determined by RT-PCR and western blotting. The terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick-end labeling (TUNEL) assay and immunohistochemical staining were assessed in renal cortex of db/db mice. RESULTS: Microarray data revealed that 7 genes show up- or down-regulated pattern and diabetic mice specifically decreased heat shock protein (Hsp) 90α expression of genes compared to control mice (diabetic mice 0.68 vs. control mice 1 relative density). Expression of Hsp90α mRNA and Hsp90 protein was significantly decreased in the renal cortex of diabetic mice. However, Hsp70 mRNA and protein expression was not changed. Apoptosis was increased in glomeruli of diabetic mice due to increased expression of cleaved caspase-3 and Bax. CONCLUSIONS: Our results suggest that Hsp 90 expression was decreased in diabetic glomeruli and decreased Hsp90 expression may mediate podocyte apoptosis in type 2 diabetic kidneys.

Aminoguanidine protects against apoptosis of retinal ganglion cells in Zucker diabetic fatty rats.

AIM: The inhibition of advanced glycation end products (AGEs) and the receptor for AGEs (RAGE) mediated downstream signaling pathways have been suggested to have retinoprotective actions in diabetic retinopathy. Herein, we examined the protective effects of aminoguanidine (AG), an AGEs inhibitor, on diabetes-induced injury of retinal ganglion cells in the Zucker diabetic fatty (ZDF) rats. MATERIALS AND METHODS: Seven-week-old male ZDF rats were treated with AG (50 mg/kg body weight) once a day orally for 13 weeks. Serum and vitreous concentration of AGEs were examined. Expressions of AGEs and its receptor (RAGE) were assessed by immunohistochemistry. Southwestern histochemistry was used to detect activated nuclear factor (NF)-κB. RESULTS: At the end of the study, vitreal levels of AGEs were significantly reduced in ZDF rats treated with AG. Similary, immunohistochemical analysis showed that AG significantly reduced the positive areas for AGEs and RAGE. Furthermore, AG strongly inhibited the loss of retinal ganglion cells by apoptosis. AG also suppressed the activation of to NF-κB. CONCLUSIONS: Our results suggest that AG has retinoprotective properties through not only direct inhibition of AGEs formation but also downregulation of NF-κB.

HyDRa: control of parameters for deterministic polishing.

Deterministic hydrodynamic polishing with HyDRa requires a precise control of polishing parameters, such as propelling air pressure, slurry density, slurry flux and tool height. We describe the HyDRa polishing system and prove how precise, deterministic polishing can be achieved in terms of the control of these parameters. The polishing results of an 84 cm hyperbolic mirror are presented to illustrate how the stability of these parameters is important to obtain high-quality surfaces.

TGF-ß suppresses the expression of genes related to mitochondrial function in lung A549 cells.

TGF-ß is a mediator of lung fibrosis and regulates the alveolar epithelial type II cell phenotype. TGF-ß can induce epithelial mesenchymal transition of idiopathic pulmonary disease and cancer metastasis. Peroxisome proliferator-activated receptor gamma co-activator 1-alpha (PGC-1 α) is a key metabolic regulator that stimulates mitochondrial biogenesis and promotes remodeling of muscle tissue to oxidative fiber-type composition. Here, we report that the induction of TGF-ß decreased mRNA expression of PGC-1α, and PGC-1 target genes, such as the transcription factors NRF-2, ERR-α, and PPAR-γ in lung epithelial A549 cells. In addition, TGF-ß led to the reduction of super oxide dismutase 2 (anti-oxidant enzyme), cytochrome C (electron transport chain in mitochondria), and MCAD (a mitochondrial ß-oxidant enzyme) in A549 cells. Together, our results suggest that TGF-ß may suppress the transcriptional activity of the genes related to mitochondrial biogenesis or function. This mechanism may provide a novel insight into the understanding of fibrosis disease.

Variant of the anastigmatic telescope with three mirrors for back focal length.

In this paper, an optical design is presented for an anastigmatic telescope with back focal length corrected with exact ray tracing to eliminate spherical, coma, and astigmatism aberrations. The telescope is formed of three conical mirrors, two of them polished on the same substratum. The optical design is divided into three stages: we began the design obtaining the Gaussian parameters in a first-order solution; posteriorly, were obtained analytically the three mirrors' asphericity in a third-order design. The final design stage consists of the implementation of the Fermat's principle, the Abbe sine condition, and the Coddington equations for the exact correction for the three aforementioned aberrations.

Renal podocyte apoptosis in Zucker diabetic fatty rats: involvement of methylglyoxal-induced oxidative DNA damage.

Methylglyoxal (MGO) is a cytotoxic metabolite produced by in-vivo glycolysis that may result in diabetic complications. The aim of this study was to determine whether MGO and oxidative stress caused apoptosis of renal podocytes in the Zucker diabetic fatty (ZDF) rat, an animal model of type 2 diabetes mellitus. Male ZDF rats aged 21 weeks developed marked hyperglycaemia with proteinuria and albuminuria. Immunohistochemical evaluation of sections of kidney demonstrated expression of MGO and 8-hydroxydeoxyguanosine (8-OHdG) in the podocytes of both normoglycaemic and diabetic rats. Podocyte apoptosis was shown through application of the TUNEL method. These findings suggest that expression of MGO and 8-OHdG is caused by hyperglycaemia, and that this expression is associated with the observed apoptosis of podocytes and is related to diabetic nephropathy.

Enhanced host immune recognition of mastitis causing Escherchia coli in CD-14 transgenic mice.

Escherchia coli causes mastitis, an economically significant disease in dairy animals. E. coli endotoxin (lipopolysaccharide, LPS) when bound by host membrane proteins such as CD-14, causes release of proinflammatory cytokines recruiting neutrophils as an early, innate immune response. Excessive proinflammatory cytokines causes tissue damage, compromising mammary function. If present, soluble CD-14 (sCD-14) might out compete membrane bound CD-14, lessening the severity of the inflammatory response. To test this hypothesis transgenic mice, expressing sCD-14 in their milk (31 to 316 microg/ml), were evaluated. A cell culture study demonstrated, in the presence of LPS, milk from transgenic mice increased secretion of cytokine IL-8 compared to milk from nontransgenic littermates (18 +/- 3 vs. 35 +/- 2 ng/mL, p < 0.001). To assess protection afforded by the transgene, glands were infused with E. coli. Recovery of bacteria showed no clear relationship between sCD14 concentration and the number of organisms recovered; however, there was a strong relationship between sCD14 concentration and edema (r(2) = 0.999, p < 0.001), as measured by weight of fluid in harvested glands. Highest expressing lines had the least edema, suggesting the presence of sCD14 had an effect on reducing the inflammatory response to E. coli, thus, possibly protecting against gland tissue damage.

Influence of jasmonic acid on endogenous gibberellin and abscisic acid in salt-stressed chard plant.

The endogenous gibberellin (GA) and abscisic acid (ABA) contents as an effect of different application times of jasmonic acid (JA) in chard seedlings exposed to salt stress were investigated. Endogenous ABA content was increased by JA treatment after NaCl treatment, rather than after JA application before NaCl treatment. JA application after NaCl treatment caused higher ABA content than treatment with 160 mM NaCl alone. Total gibberellin content decreased after NaCl stress, but NaCl-reduction in total GA contents counteracted by exogenous JA. Total endogenous GA contents were increased in JA treatment after NaCl and were highest at 24 hr of JA application before NaCl exposure. JA treatment promoted the increase of dry weight compared to chard plant exposed to 160 mM NaCl. Thus, JA presumably induces gibberellin biosynthesis showing the promotion of growth and dry weight of chard plants under salt stress.

Generation and characterization of recombinant ScFv antibodies detecting Eimeria acervulina surface antigens.

In our previous attempt to generate monoclonal antibodies (MAbs) against coccidia parasites that more accurately reflect the natural avian humoral immune response, we produced two chicken B-cell hybridomas, 5D11 and 2-1. While both cell lines secreted antibodies reactive with sporozoites of Eimeria acervulina, they were produced in yields too low to conduct meaningful in vivo studies. To circumvent this problem, we produced four single chain variable fragment (scFv) antibodies from the V(H) and V(L) genes of hybridomas 5D11 and 2-1. The concentration of these recombinant antibodies expressed in E. coli and purified to homogeneity was 5-6 mg/L. Three of the antibodies exhibited antigen binding specificity to Eimeria surface antigens equivalent to that of the native MAbs. Nucleotide sequence analysis of the V(L) genes from hybridomas 5D11 and 2-1 and genomic DNA revealed vestiges of gene conversion with V(lambda) pseudogenes. These recombinant scFv antibodies will prove useful for further characterization of natural Eimeria surface antigens as potential vaccine candidates.

Production and characterization of monoclonal antibodies detecting chicken interleukin-2 and the development of an antigen capture enzyme-linked immunosorbent assay.

Eleven monoclonal antibodies (mAbs) which are specific for chicken interleukin-2 (chIL-2) were produced and characterized by enzyme-linked immunosorbent assay (ELISA), Western blotting and neutralizing assays. These mAbs were used to develop a mAb-based antigen capture ELISA specific for chicken IL-2 detection. Anti-IL-2 mAbs bound specifically to E. coli-derived rchIL-2 in ELISA and identified a 16kDa IL-2 polypeptide band in Western blot. Several mAbs were shown to neutralize the biological activities of both rchIL-2 and native chicken IL-2 as measured by concanavalin A (ConA)-induced lymphocyte proliferation assay, IL-2 bioassay, and natural killer cell assay. Among the neutralizing mAbs, the mAb chIL-2/11 was most potent in neutralizing IL-2 activity. To develop a sensitive ELISA for the detection of chicken IL-2, an antigen capture ELISA was developed using the mAb chIL-2/16 as the antigen capture antibody and rabbit anti-IL-2 peptide antibody as the detection antibody. Using the mAb-based antigen capture ELISA, significant correlation between the level of IL-2 detected in bioassays and in ELISA was observed. These results showed that the mAb-based antigen capture ELISA is less time-consuming and more reliable compared to a conventional IL-2 bioassay for chicken IL-2. These neutralizing mAbs will facilitate basic immunobiological studies of the role of IL-2 in normal and disease states in chickens.

Phenotypic variation of a Thr704Met mutation in skeletal sodium channel gene in a family with paralysis periodica paramyotonica.

OBJECTIVES: Patients with paralysis periodica paramyotonica exhibit a clinical syndrome with characteristics of both hyperkalaemic periodic paralysis and paramyotonia congenita. In several types of periodic paralysis associated with hyperkalaemia, mutations in the skeletal muscle sodium channel (SCN4A) gene have been previously reported. Phenotypic variations of mutations in SCN4A, however, have not been described yet. The present study aimed to evaluate genetic variations in a family with clinical and electrophysiological characteristics of paralysis periodica paramyotonia. METHODS: Seven members of a family affected with symptoms of paralysis periodica paramyotonia were studied by electrophysiological and genetic analyses. There were increased serum potassium concentrations in four members during paralytic attacks induced by hyperkalaemic periodic paralysis provocation tests. Short exercise tests before and after cold immersion were carried out in four patients to distinguish electrophysiological characteristics of hyperkalaemic periodic paralysis and paramyotonia. Sequencing analyses of SCN4A were performed on one patient and a normal control to identify polymorphisms. Restriction fragment length polymorphism (RFLP) analysis was then performed at the identified polymorphic sites. RESULTS: Electrophysiological studies showed both exercise sensitivity and temperature sensitivity. Compound motor action potential (CMAP) amplitudes were decreased (7.3%-28.6%) after short exercise tests. The CMAP amplitudes were even more severely decreased (21.7%-56.5%) in short exercise tests after cold exposure. Three polymorphic sites, Gln371Glu, Thr704Met, and Aspl376Asn were identified in SCN4A. RFLP analyses showed that all affected patients carried the Thr704Met mutation, whereas unaffected family members and a normal control did not. CONCLUSION: Phenotypic variation of the Thr704Met mutation, which was previously reported in patients with hyperkalaemic periodic paralysis, is described in a family affected with paralysis periodica paramyotonia.

Analysis of chromosome aberrations in nuclear-power-plant workers considering the lifetime of lymphocytes.

PURPOSE: To analyse chromosome aberrations in nuclear-power-plant workers taking account of the mean lifetime of lymphocytes (MLTL). MATERIALS AND METHODS: Analysis of chromosome aberrations was performed on peripheral lymphocytes from 395 nuclear-power-plant workers and 135 controls. An equivalent acute dose (EAD) was calculated utilizing MLTL values of either 4.3 or 10 years. RESULTS: Using an MLTL value of 10 years produced an EAD range of 0.O1 mSv -182mSv(mean 46.6mSv), while using an MLTL, of 4.3 years produced results ranging from 0.01 mSv to 86.2 mSv (mean 23.4 mSv). A significant increase of chromosome-type exchange by the equivalent acute dose was observed using an MLTL of either 10 or 4.3 years when including the control in the analysis, but a significant increase was not seen when only the exposed was considered. A significant increase of chromosome-type deletion by EAD was seen even when only the exposed group was considered. CONCLUSIONS: EAD values based on an MLTL of either 4.3 or 10 years, as well as cumulative dose, showed no significant association with chromosome aberrations, when radiation workers only were analysed. The narrow dose range examined in this study might have contributed to this finding.