Clinical Significance of Heparanase Splice Variant (T5) in Renal Cell Carcinoma: Evaluation by a Novel T5-Specific Monoclonal Antibody

T5 is a novel splice variant of heparanase, an endo-β-D-glucuronidase capable of cleaving heparan sulfate side chains at a limited number of sites. T5 splice variant is endowed with pro-tumorigenic properties, enhancing cell proliferation, anchorage independent growth and tumor xenograft development despite lack of heparan sulfate-degrading activity typical of heparanase. T5 is over expressed in the majority of human renal cell carcinoma biopsies examined, suggesting that this splice variant is clinically relevant. T5 is thought to assume a distinct three-dimensional conformation compared with the wild type heparanase protein. We sought to exploit this presumed feature by generating monoclonal antibodies that will recognize the unique structure of T5 without, or with minimal recognition of heparanase, thus enabling more accurate assessment of the clinical relevance of T5. We provide evidence that such a monoclonal antibody, 9c9, preferentially recognizes T5 compared with heparanase by ELISA, immunoblotting and immunohistochemistry. In order to uncover the clinical significance of T5, a cohort of renal cell carcinoma specimens was subjected to immunostaining applying the 9c9 antibody. Notably, T5 staining intensity was significantly associated with tumor size (p = 0.004) and tumor grade (p = 0.02). Our results suggest that T5 is a functional, pro-tumorigenic entity.     

Type II collagen antibody response is enriched in the synovial fluid of rheumatoid joints and directed to the same major epitopes as in collagen induced arthritis in primates and mice

Introduction

Antibodies towards type II collagen (CII) are detected in patients with rheumatoid arthritis (RA) and in non-human primates and rodents with collagen induced arthritis (CIA). We have previously shown that antibodies specific for several CII-epitopes are pathogenic using monoclonal antibodies from arthritic mice, although the role of different anti-CII epitopes has not been investigated in detail in other species. We therefore performed an inter-species comparative study of the autoantibody response to CII in patients with RA versus monkeys and mice with CIA.

Methods

Analysis of the full epitope repertoire along the disease course of CIA was performed using a library of CII triple-helical peptides. The antibody responses to the major CII epitopes were analyzed in sera and synovial fluid from RA patients, and in sera from rhesus monkeys (Macaca mulatta), common marmosets (Callithrix jacchus) and mice.

Results

Many CII epitopes including the major C1, U1, and J1 were associated with established CIA and arginine residues played an important role in the anti-CII antibody interactions. The major epitopes were also recognized in RA patients, both in sera and even more pronounced in synovial fluid: 77% of the patients had antibodies to the U1 epitope. The anti-CII immune response was not restricted to the anti-citrulline protein antibodies (ACPA) positive RA group.

Conclusion

CII conformational dependent antibody responses are common in RA and are likely to originate from rheumatoid joints but did not show a correlation with ACPA response. Importantly, the fine specificity of the anti-CII response is similar with CIA in monkeys and rodents where the recognized epitopes are conserved and have a major pathogenic role. Thus, anti-CII antibodies may both contribute to, as well as be the consequence of, local joint inflammation.     

Monoterpene content in Origanum syriacum as affected by environmental conditions and flowering

A high content of essential oil in the foliage of two chemotypes of field-grown Origanum syriacum L. was found between August and October. The content of phenolic monoterpenes in the essential oil was low from November to January, while the content of p -cymene was high. The effect of environmental conditions and flowering on the two chemotypes was similar. Controlled experiments revealed that under long photoperiods the content of essential oils in the foliage and of phenolic monoterpenes in the oil increased, while that of p -cymene decreased. A similar effect was observed when an 8 h natural light day was extended by 8 h low-intensity incandescent light. When the 16 h day was composed of 12 h natural light, the effect on oil content and composition was intensified. Under short-day photoperiods, increasing temperature or light intensity enhanced the relative content of p -cymene, while that of phenolic monoterpenes and the γ-terpinene generally decreased. Flowering decreased the essential oil content in the leaves and the relative content of the phenolic monoterpenes in the essential oil. Since environmental conditions influence flowering, they had both direct and indirect effects on the essential oil content and composition.     

Monocytes are essential for inhibition of synovial T-cell glucocorticoid-mediated apoptosis in rheumatoid arthritis

Introduction  

Rheumatoid arthritis (RA) is characterized by synovial inflammation with local accumulation of mononuclear cells such as macrophages and lymphocytes. We previously demonstrated that intra-articular glucocorticoids decrease the synovial tissue (ST) T-cell population and therefore aimed to investigate whether this is mediated through modulation of apoptosis.     

Intraspecific variation of Chiliadenus iphionoides essential oil in Israel

Chiliadenus iphionoides (Asteraceae), a shrub endemic to the Mediterranean region and widespread throughout Israel, is used in the traditional eastern Mediterranean medicine. Although recent research confirmed its pharmacological potential, C. iphionoides essential oil has not been adequately characterized chemically. Essential-oil samples were collected from representative wild populations throughout Israel and characterized by GC/MS analysis. Considerable interpopulation variation was found for the composition of the essential oils. Multivariate analysis showed a significant correlation between the chemical composition and the geographic location, with three main chemotypes identified.     

Quantitative study of optical and mechanical bone status using multispectral photoacoustics

Osteoporosis is a major public health problem worldwide. Here, we present a quantitative multispectral photoacoustic method for the evaluation of bone pathologies which has significant advantages over pure ultrasonic or pure optical methods as it provides both molecular information and bone mechanical status. This is enabled via a simultaneous measurement of the bone's optical properties as well as the speed of sound and ultrasonic attenuation in the bone. To test the method's quantitative predictions, a combined ultrasonic and photoacoustic system was developed. Excitation was performed optically via a portable triple laser‐diode system and acoustically via a single element transducer. Additional dual transducers were used for detecting the acoustic waves that were generated by the two modalities. Both temporal and spectral parameters were compared between different excitation wavelengths and measurement modalities. Short photoacoustic excitation wavelengths allowed sensing of the cortical layer while longer wavelengths produced results which were compatible with the quantitative ultrasound measurements.

     

Personal receptor repertoires: olfaction as a model

ABSTRACT: BACKGROUND: Information on nucleotide diversity along completely sequenced human genomes has increased tremendously over the last few years. This makes it possible to reassess the diversity status of distinct receptor proteins in different human individuals. To this end, we focused on the complete inventory of human olfactory receptor coding regions as a model for personal receptor repertoires. RESULTS: By performing data-mining from public and private sources we scored genetic variations in 413 intact OR loci, for which one or more individuals had an intact open reading frame. Using 1000 Genomes Project haplotypes, we identified a total of 4069 full-length polypeptide variants encoded by these OR loci, average of ~10 per locus, constituting a lower limit for the effective human OR repertoire. Each individual is found to harbor as many as 600 OR allelic variants, ~50% higher than the locus count. Because OR neuronal expression is allelically excluded, this has direct effect on smell perception diversity of the species. We further identified 244 OR segregating pseudogenes (SPGs), loci showing both intact and pseudogene forms in the population, twenty-six of which are annotatively "resurrected" from a pseudogene status in the reference genome. Using a custom SNP microarray we validated 150 SPGs in a cohort of 468 individuals, with every individual genome averaging 36 disrupted sequence variations, 15 in homozygote form. Finally, we generated a multi-source compendium of 63 OR loci harboring deletion Copy Number Variations (CNVs). Our combined data suggest that 271 of the 413 intact OR loci (66%) are affected by nonfunctional SNPs/indels and/or CNVs. CONCLUSIONS: These results portray a case of unusually high genetic diversity, and suggest that individual humans have a highly personalized inventory of functional olfactory receptors, a conclusion that might apply to other receptor multigene families.     

The Taxonomy and Phylogeny of Echinometra (Camarodonta: Echinometridae) from the Red Sea and Western Indian Ocean

The number of valid species in the genus Echinometra (Echinodermata, Echinoidea) and their associated identification keys have been debated in the scientific literature for more than 180 years. As the phylogeny and dispersal patterns of these species have been widely used as a prominent model for marine speciation, new insights into their taxonomy have the potential to deepen our understanding of marine speciation processes. In this study we examine Echinometra taxonomy, combining morphology and molecular tools. We present the taxonomy and phylogeny of Red Sea and Western Indian Ocean Echinometra. The currently available morphological keys were found to be limited in their ability to delineate all species within this genus. Nonetheless, morphological similarities between the Red Sea and Western Indian Ocean populations were high, and delimited them from the other species. These latter populations together formed a monophyletic clade, genetically distant from any of the other Echinometra species by more than 3%. Combining both traditional taxonomy and molecular evidence, we found that these populations were neither Echinometra mathaei nor E. oblonga, as previously considered. The morphological discrepancies of these populations, and their genetic divergence from the other Echinometra species, suggest that they should be considered as a new Echinometra species.