Codistribution of collagen types IV and AB2 in basement membranes and mesangium of the kidney. an immunoferritin study of ultrathin frozen sections

Affinity-purified rabbit antibodies specific for collagen types I, III, AB2 and for a partially characterized type IV collagen derived from a murine tumor were used to study the distribution of collagens in the normal mouse kidney. Immunofluorescence staining of conventional frozen sections demonstrated that types I and III were present in bundles around large vessels and in fibers surrounding glomeruli and tubules, whereas types IV and AB2 were distributed in a linear fashion along basement membranes of tubules, glomeruli, and Bowman's capsule and in the mesangial stalk. The distribution of types IV nd AB2 was examined at the ultrastructural level by staining of 600- to 800-A thick frozen sections with a three-stage procedure employing specific collagen antibodies, biotinyl sheep antirabbit IgG, and avidin-ferritin conjugates. Labeling by this procedure demonstrated codistribution of types AB2 and the putative type IV in all three basement membranes. In addition, mesangial matrix was shown to contain both of these collagen types. These results support recent biochemical evidence of collagen heterogeneity in basement membranes, and also support the concept of a structural relationship between mesangial matrix and glomerular basement membranes.     

Non-indigenous terrestrial vertebrates in Israel and adjacent areas

We investigated characteristics of established non-indigenous (ENI) terrestrial vertebrates in Israel and adjacent areas, as well as attributes of areas they occupy. Eighteen non-indigenous birds have established populations in this region since 1850. A database of their attributes was compiled, analyzed, and compared to works from elsewhere. Most ENI bird species are established locally; a few are spreading or widespread. There has been a recent large increase in establishment. All ENI birds are of tropical origin, mostly from the Ethiopian and Oriental regions; the main families are Sturnidae, Psittacidae, Anatidae, and Columbidae. Most species have been deliberately brought to Israel in captivity and subsequently released or escaped. Most of these birds are commensal with humans to some degree, are not typically migratory, and have mean body mass larger than that of the entire order. ENI birds are not distributed randomly. There are centers in the Tel-Aviv area and along the Rift Valley, which is also a corridor of spread. Positive correlations were found between ENI bird richness and mean annual temperature and urbanization. Mediterranean forests and desert regions have fewer ENI species than expected. Apart from birds we report on non-indigenous species of reptiles (2) and mammals (2) in this region.     

Damage functions and thermal requirements of Meloidogyne javanica and Meloidogyne incognita on watermelon

The relationship between the initial (P_i) and final (P_f) population densities of Meloidogyne javanica and yield of watermelon, Citrullus lanatus, cv. Sugar Baby were determined in pot and field experiments. In the pots, the maximum reproduction rate of the nematode was 14, and the equilibrium density was 49 400 eggs/100 cm³ of soil. Yield data represented as fresh top weight fitted the Seinhorst damage function (P < 0.001, R² = 0.7), and the minimum relative yield (m) was 0.65 at P_i ≥ 3200 eggs/100 cm³ of soil and the tolerance limit (T) 74 eggs/100 cm³. In the field experiments (2011 and 2012), the maximum reproduction rate was 73 and 70, and the equilibrium density 32 and 35 second‐stage juveniles (J2)/100 cm³ soil. Yield data represented as fruit weight fitted the Seinhorst damage function in 2011 (P < 0.001, R² = 0.92) and the m‐ and T‐values were 0.63 and 20 J2/100 cm³ of soil, respectively. Meloidogyne incognita and M. javanica needed similar length of time for development to egg‐laying females and life cycle completion at 24.4°C.     

Non-indigenous land and freshwater gastropods in Israel

Few comprehensive works have investigated non-indigenous snails and slugs as a group. We compiled a database of non-indigenous gastropods in Israel to explore how they arrived and spread, characteristics of their introduction, and their biological traits. Fifty-two species of introduced gastropods are known from Israel (of which nine species subsequently went extinct): 19 species of freshwater snails and 33 species of terrestrial gastropods. The majority of these species are found only in human-dominated habitats. Most of those found in natural habitats are aquatic species. Most snails are introduced unintentionally from various parts of the Holoarctic region, reaching Israel as stowaways with horticultural imports and the aquarium trade, but some are brought intentionally to be used as pets or for food. Because the study of this group in Israel is very limited, information regarding their distribution in the country and their effects on other species is incomplete. Though only nine species of non-indigenous snails have been found to date in natural habitats, some of these are very abundant. More information and research is required to enable effective management schemes.     

Circulating surfactant protein -D is low and correlates negatively with systemic inflammation in early, untreated rheumatoid arthritis

Introduction

Surfactant protein D (SP-D) is a collectin with immuno-regulatory functions, which may depend on oligomerization. Anti-microbial and anti-inflammatory properties have been attributed to multimeric SP-D variants, while trimeric subunits per se have been suggested to enhance inflammation. Previously, we reported low circulating SP-D in early rheumatoid arthritis (RA), and the present investigation aims to extend these data by serial SP-D serum measurements, studies on synovial fluid, SP-D size distribution and genotyping in patients with early RA.

Methods

One-hundred-and-sixty disease-modifying antirheumatic drug (DMARD) naïve RA patients with disease duration less than six months were studied prospectively for four years (CIMESTRA (Ciclosporine, Methotrexate, Steroid in RA) trial) including disease activity measures (C-reactive protein, joint counts and Health Assessment Questionnaire (HAQ) score), autoantibodies, x-ray findings and SP-D. SP-D was quantified by enzyme-linked immunosorbent assay (ELISA) and molecular size distribution was assessed by gel filtration chromatography. Further, SP-D Met11Thr single nucleotide polymorphism (SNP) analysis was performed.

Results

Serum SP-D was significantly lower in RA patients at baseline compared with healthy controls (P < 0.001). SP-D increased slightly during follow-up (P < 0.001), but was still subnormal at four years after adjustment for confounders (P < 0.001). SP-D in synovial fluid was up to 2.5-fold lower than in serum. While multimeric variants were detected in serum, SP-D in synovial fluid comprised trimeric subunits only. There were no significant associations between genotype distribution and SP-D. Baseline SP-D was inversely associated to CRP and HAQ score. A similar relationship was observed regarding temporal changes in SP-D and CRP (zero to four years). SP-D was not associated to x-ray findings.

Conclusions

This study confirms that circulating SP-D is persistently subnormal in early and untreated RA despite a favourable therapeutic response obtained during four years of follow-up. SP-D correlated negatively to disease activity measures, but was not correlated with x-ray progression or SP-D genotype. These observations suggest that SP-D is implicated in RA pathogenesis at the protein level. The exclusive presence of trimeric SP-D in affected joints may contribute to the maintenance of joint inflammation.

Trial registration

(j.nr NCT00209859).     

Effect of Ca2+, Ba2+, and Sr2+ on alginate microbeads

Microcapsules of alginate cross-linked with divalent ions are the most common system for cell immobilization. In this study, we wanted to characterize the effect of different alginates and cross-linking ions on important microcapsule properties. The dimensional stability and gel strength increased for high-G alginate gels when exchanging the traditional Ca2+ ions with Ba2+. The use of Ba2+ decreased the size of alginate beads and reduced the permeability to immunoglobulin G. Strontium gave gels with characteristics lying between calcium and barium. Interestingly, high-M alginate showed an opposite behavior in combination with barium and strontium as these beads were larger than beads of calcium-alginate and tended to swell more, also resulting in increased permeability. Binding studies revealed that different block structures in the alginate bind the ions to a different extent. More specifically, Ca2+ was found to bind to G- and MG-blocks, Ba2+ to G- and M-blocks, and Sr2+ to G-blocks solely.     

C6-oxidation and c5-epimerization of locust bean galactomannan studied by high field NMR and circular dichroism

Galactose depleted locust bean gum was selectively oxidized in C(6) position and epimerized with mannuronan C(5)-epimerases to obtain the corresponding artificial uronanes. These new pseudo-alginates were characterized by NMR spectroscopy and circular dichroism (CD). Specifically, 1D and 2D NMR techniques allowed the degree of epimerization, the distribution of mannuronic and guluronic acid residues in the polysaccharidic chain, and the average G block length to be determined. In addition, NMR diffusion experiments showed that the epimerization reaction did not significantly degrade the polysaccharidic chains. Circular dichroism was used to investigate the kinetics of the epimerization reaction and to evidence the specific interaction between the epimerized locust bean samples with Ca(II) ions in dilute solution. All of the samples considered in this study form wall to wall gels in concentrated polymer solutions.     

Infantile convulsions with paroxysmal dyskinesia (ICCA syndrome) and copy number variation at human chromosome 16p11

Background

Benign infantile convulsions and paroxysmal dyskinesia are episodic cerebral disorders that can share common genetic bases. They can be co-inherited as one single autosomal dominant trait (ICCA syndrome); the disease ICCA gene maps at chromosome 16p12-q12. Despite intensive and conventional mutation screening, the ICCA gene remains unknown to date. The critical area displays highly complicated genomic architecture and is the site of deletions and duplications associated with various diseases. The possibility that the ICCA syndrome is related to the existence of large-scale genomic alterations was addressed in the present study.

Methodology/Principal Findings

A combination of whole genome and dedicated oligonucleotide array comparative genomic hybridization coupled with quantitative polymerase chain reaction was used. Low copy number of a region corresponding to a genomic variant (Variation_7105) located at 16p11 nearby the centromere was detected with statistical significance at much higher frequency in patients from ICCA families than in ethnically matched controls. The genomic variant showed no apparent difference in size and copy number between patients and controls, making it very unlikely that the genomic alteration detected here is ICCA-specific. Furthermore, no other genomic alteration that would directly cause the ICCA syndrome in those nine families was detected in the ICCA critical area.

Conclusions/Significance

Our data excluded that inherited genomic deletion or duplication events directly cause the ICCA syndrome; rather, they help narrowing down the critical ICCA region dramatically and indicate that the disease ICCA genetic defect lies very close to or within Variation_7105 and hence should now be searched in the corresponding genomic area and its surrounding regions.