Hyaluronan synthases (HAS1-3) and hyaluronidases (HYAL1-2) in the accumulation of hyaluronan in endometrioid endometrial carcinoma

<p>Abstract</p> <p>Background</p> <p>Hyaluronan accumulation correlates with the degree of malignancy in many solid tumor types, including malignant endometrial carcinomas. To elucidate the mechanism of hyaluronan accumulation, we examined the expression levels of the hyaluronan synthases (<it>HAS1</it>, <it>HAS2 </it>and <it>HAS3</it>) and hyaluronidases (<it>HYAL1 </it>and <it>HYAL2</it>), and correlated them with hyaluronan content and HAS1-3 immunoreactivity.</p> <p>Methods</p> <p>A total of 35 endometrial tissue biopsies from 35 patients, including proliferative and secretory endometrium (n = 10), post-menopausal proliferative endometrium (n = 5), complex atypical hyperplasia (n = 4), grade 1 (n = 8) and grade 2 + 3 (n = 8) endometrioid adenocarcinomas were divided for gene expression by real-time RT-PCR, and paraffin embedded blocks for hyaluronan and HAS1-3 cytochemistry.</p> <p>Results</p> <p>The mRNA levels of <it>HAS1-3 </it>were not consistently changed, while the immunoreactivity of all HAS proteins was increased in the cancer epithelium. Interestingly, <it>HAS3 </it>mRNA, but not HAS3 immunoreactivity, was increased in post-menopausal endometrium compared to normal endometrium (p = 0.003). The median of <it>HYAL1 </it>mRNA was 10-fold and 15-fold lower in both grade 1 and grade 2+3 endometrioid endometrial cancers, as compared to normal endometrium (p = 0.004-0.006), and post-menopausal endometrium (p = 0.002), respectively. <it>HYAL2 </it>mRNA was also reduced in cancer (p = 0.02) and correlated with <it>HYAL1</it> (r = 0.8, p = 0.0001). There was an inverse correlation between <it>HYAL1 </it>mRNA and the epithelial hyaluronan staining intensity (r = -0.6; P = 0.001).</p> <p>Conclusion</p> <p>The results indicated that <it>HYAL1 </it>and <it>HYAL2 </it>were coexpressed and significantly downregulated in endometrioid endometrial cancer and correlated with the accumulation of hyaluronan. While immunoreactivity for HASs increased in the cancer cells, tumor mRNA levels for <it>HAS</it>s were not changed, suggesting that reduced turnover of HAS protein may also have contributed to the accumulation of hyaluronan.</p

Expression of Hyaluronan Synthases (HAS1–3) and Hyaluronidases (HYAL1–2) in Serous Ovarian Carcinomas: Inverse Correlation between HYAL1 and Hyaluronan Content

<p>Abstract</p> <p>Background</p> <p>Hyaluronan, a tumor promoting extracellular matrix polysaccharide, is elevated in malignant epithelial ovarian tumors, and associates with an unfavorable prognosis. To explore possible contributors to the accumulation of hyaluronan, we examined the expression of hyaluronan synthases (<it>HAS1</it>, <it>HAS2 </it>and <it>HAS3</it>) and hyaluronidases (<it>HYAL1 </it>and <it>HYAL2</it>), correlated with hyaluronidase enzyme activity hyaluronan content and HAS1–3 immunoreactivity.</p> <p>Methods</p> <p>Normal ovaries (n = 5) and 34 serous epithelial ovarian tumors, divided into 4 groups: malignant grades 1+2 (n = 10); malignant grade 3 (n = 10); borderline (n = 4) and benign epithelial tumors (n = 10), were analyzed for mRNA by real-time RT-PCR and compared to hyaluronidase activity, hyaluronan staining, and HAS1–3 immunoreactivity in tissue sections of the same specimens.</p> <p>Results</p> <p>The levels of <it>HAS2 </it>and <it>HAS3 </it>mRNA (<it>HAS1 </it>was low or absent), were not consistently increased in the carcinomas, and were not significantly correlated with HAS protein or hyaluronan accumulation in individual samples. Instead, the median of <it>HYAL1 </it>mRNA level was 69% lower in grade 3 serous ovarian cancers compared to normal ovaries (P = 0.01). The expression of <it>HYAL1</it>, but not <it>HYAL2</it>, significantly correlated with the enzymatic activity of tissue hyaluronidases (r = 0.5; P = 0.006). An inverse correlation was noted between <it>HYAL1 </it>mRNA and the intensity of hyaluronan staining of the corresponding tissue sections (r = -0.4; P = 0.025).</p> <p>Conclusion</p> <p>The results indicate that in serous epithelial ovarian malignancies <it>HAS </it>expression is not consistently elevated but <it>HYAL1 </it>expression is significantly reduced and correlates with the accumulation of hyaluronan. (233 words)</p

Single-Molecule Unbinding Forces between the Polysaccharide Hyaluronan and Its Binding Proteins

The extracellular polysaccharide hyaluronan (HA) is ubiquitous in all vertebrate tissues, where its various functions are encoded in the supramolecular complexes and matrices that it forms with HA-binding proteins (hyaladherins). In tissues, these supramolecular architectures are frequently subjected to mechanical stress, yet how this affects the intermolecular bonding is largely unknown. Here, we used a recently developed single-molecule force spectroscopy platform to analyze and compare the mechanical strength of bonds between HA and a panel of hyaladherins from the Link module superfamily, namely the complex of the proteoglycan aggrecan and cartilage link protein, the proteoglycan versican, the inflammation-associated protein TSG-6, the HA receptor for endocytosis (stabilin-2/HARE), and the HA receptor CD44. We find that the resistance to tensile stress for these hyaladherins correlates with the size of the HA-binding domain. The lowest mean rupture forces are observed for members of the type A subgroup (i.e., with the shortest HA-binding domains; TSG-6 and HARE). In contrast, the mechanical stability of the bond formed by aggrecan in complex with cartilage link protein (two members of the type C subgroup, i.e., with the longest HA-binding domains) and HA is equal or even superior to the high affinity streptavidin,biotin bond. Implications for the molecular mechanism of unbinding of HA, hyaladherin bonds under force are discussed, which underpin the mechanical properties of HA, hyaladherin complexes and HA-rich extracellular matrices

Structure of the regulatory hyaluronan binding domain in the inflammatory leukocyte homing receptor CD44

Adhesive interactions involving CD44, the cell surface receptor for hyaluronan, underlie fundamental processes such as inflammatory leukocyte homing and tumor metastasis. Regulation of such events is critical and appears to be effected by changes in CD44 N-glycosylation that switch the receptor "on" or "off" under appropriate circumstances. How altered glycosylation influences binding of hyaluronan to the lectin-like Link module in CD44 is unclear, although evidence suggests additional flanking sequences peculiar to CD44 may be involved. Here we show using X-ray crystallography and NMR spectroscopy that these sequences form a lobular extension to the Link module, creating an enlarged HA binding domain and a formerly unidentified protein fold. Moreover, the disposition of key N-glycosylation sites reveals how specific sugar chains could alter both the affinity and avidity of CD44 HA binding. Our results provide the necessary structural framework for understanding the diverse functions of CD44 and developing novel therapeutic strategies

A preformed basal lamina alters the metabolism and distribution of hyaluronan in epidermal keratinocyte "organotypic" cultures grown on collagen matrices

A rat epidermal keratinocyte (REK) line which exhibits histodifferentiation nearly identical to the native epidermis when cultured at an air–liquid interface was used to study the metabolism of hyaluronan, the major intercellular macromolecule present in basal and spinous cell layers. Two different support matrices were used: reconstituted collagen fibrils with and without a covering basal lamina previously deposited by canine kidney cells. REKs formed a stratified squamous, keratinized epithelium on both support matrices. Hyaluronan and its receptor, CD44, colocalized in the basal and spinous layers similar to their distribution in the native epidermis. Most (approximately 75%) of the hyaluronan was retained in the epithelium when a basal lamina was present while most (approximately 80%) diffused out of the epithelium in its absence. While REKs on the two matrices synthesized hyaluronan at essentially the same rate, catabolism of this macromolecule was much higher in the epithelium on the basal lamina (half-life approximately 1 day, similar to its half-life in native human epidermis). The formation of a true epidermal compartment in culture bounded by the cornified layer on the surface and the basal lamina subjacent to the basal cells provides a good model within which to study epidermal metabolism.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/42232/1/418-113-4-265_s004180000128.pd

APLIKASI ANALISIS KLASTER DAN INDEKS TRIX UNTUK MENGKAJI VARIABILITAS STATUS TROFIK DI TELUK PEGAMETAN, SINGARAJA, BALI

Tren variabilitas status trofik adalah informasi dasar pengelolaan ekosistem perairan dalam kaitannya dengan tingkat eutrofikasi. Tujuan penelitian ini adalah menentukan tren variabilitas status trofik baik secara spasial dan temporal sehingga dapat diterapkan dalam konsep pengelolaan perairan. Selama tiga bulan penelitian (Agustus-Oktober 2014), penelitian ini telah berhasil mengumpulkan data dari 48 titik pengamatan baik saat pasang maupun surut yang bertepatan saat bulan purnama maupun gelap. Data yang dikumpulkan lalu dianalisis secara statistik (analisis klaster) dan penentuan indeks TRIX. Hasil analisis secara spasial menunjukkan bahwa lokasi penelitian terbagi menjadi dua wilayah yakni: K1 untuk kawasan pesisir teluk, K2 untuk tengah, dan ujung teluk dengan nilai masing-masing adalah 4,97±0,92 dan 5,51±0,90. Analisis secara temporal menghasilkan dua kelompok yakni pada bulan Agustus (A1) dan September-Oktober (A2) dengan nilai indeks TRIX masing-masing 4,28±0,99 dan 5,78±0,27. Berdasarkan analisis klaster dan indeks TRIX telah menunjukkan secara jelas bahwa kondisi kawasan klaster K1 lebih baik dibandingkan kawasan klaster K2. Kawasan K1 masih dapat dikembangkan untuk budidaya laut keramba jaring apung (KJA), namun dalam perkembangannya, budidaya laut secara terintegrasi berbasis integrated multi-trophic aquaculture (IMTA) dapat diterapkan guna menjaga daya dukung lingkungan perairan Teluk Pegametan. Tren secara temporal menunjukkan pola yang mengikuti fluktuasi air atau pasang surut, serta jadwal kematian massal ikan setiap tahunnya dan berbeda nyata pada saat Agustus hingga September-Oktober. Hasil penelitian ini diharapkan memberikan informasi tentang kondisi perairan Teluk Pegametan sehingga dapat dilakukan pengelolaan kawasan yang lebih baik untuk kegiatan budidaya laut yang berkelanjutan

APLIKASI ANALISIS KLASTER DAN INDEKS TRIX UNTUK MENGKAJI VARIABILITAS STATUS TROFIK DI TELUK PEGAMETAN, SINGARAJA, BALI

Tren variabilitas status trofik adalah informasi dasar pengelolaan ekosistem perairan dalam kaitannya dengan tingkat eutrofikasi. Tujuan penelitian ini adalah menentukan tren variabilitas status trofik baik secara spasial dan temporal sehingga dapat diterapkan dalam konsep pengelolaan perairan. Selama tiga bulan penelitian (Agustus-Oktober 2014), penelitian ini telah berhasil mengumpulkan data dari 48 titik pengamatan baik saat pasang maupun surut yang bertepatan saat bulan purnama maupun gelap. Data yang dikumpulkan lalu dianalisis secara statistik (analisis klaster) dan penentuan indeks TRIX. Hasil analisis secara spasial menunjukkan bahwa lokasi penelitian terbagi menjadi dua wilayah yakni: K1 untuk kawasan pesisir teluk, K2 untuk tengah, dan ujung teluk dengan nilai masing-masing adalah 4,97±0,92 dan 5,51±0,90. Analisis secara temporal menghasilkan dua kelompok yakni pada bulan Agustus (A1) dan September-Oktober (A2) dengan nilai indeks TRIX masing-masing 4,28±0,99 dan 5,78±0,27. Berdasarkan analisisklaster dan indeks TRIX telah menunjukkan secara jelas bahwa kondisi kawasan klaster K1 lebih baik dibandingkan kawasan klaster K2. Kawasan K1 masih dapat dikembangkan untuk budidaya laut keramba jaring apung (KJA), namun dalam perkembangannya, budidaya laut secara terintegrasi berbasis integrated multi-trophic aquaculture (IMTA) dapat diterapkan guna menjaga daya dukung lingkungan perairan Teluk Pegametan. Tren secara temporal menunjukkan pola yang mengikuti fluktuasi air atau pasang surut, serta jadwal kematian massal ikan setiap tahunnya dan berbeda nyata pada saat Agustus hingga September-Oktober. Hasil penelitian ini diharapkan memberikan informasi tentang kondisi perairan Teluk Pegametan sehingga dapat dilakukan pengelolaan kawasan yang lebih baik untuk kegiatan budidaya laut yang berkelanjutan

Mindsets and Failures : Neural Differences in Reactions to Mistakes among 2nd Grade Finnish Girls

Mindsets have been identified as an important factor in explaining learning differences among students. Growth mindset students have been shown to recover from mistakes easier than fixed mindset students, and recent neuroscientific research has shown differences in the brain’s event-related potentials to errors in fixed and growth mindset participants. The purpose of this study was to examine and evaluate these differences in the Finnish elementary school context. To achieve this, event-related potentials of five fixed and five growth mindset 8-9-year-old female students were recorded during a go/no-go task. Differences between the two groups emerged, however, they were different from the results of some previous studies in the field. These findings are discussed in the light of earlier neuroscientific research related to mindsets, including limitations and suggestions for future research in the field.Peer reviewe

Decline after immobilisation and recovery after remobilisation of synovial fluid IL1, TIMP, and chondroitin sulphate levels in young beagle dogs

OBJECTIVE: To monitor the concentration of markers of cartilage and synovium metabolism in the knee (stifle) joint synovial fluid of young beagles subjected to immobilisation and subsequent remobilisation.METHODS: The right hind limb of 17 dogs was immobilised in flexion for 11 weeks. Simultaneously, the contralateral left knee was exposed to increased weight bearing. The remobilisation period lasted 50 weeks. Litter mates served as controls. The concentration in joint lavage fluid of interleukin 1 (IL1) was measured by immunoassay, the activity of phospholipase A2 (PLA2) was determined by an extraction method, chondroitin sulphate (CS) concentration by precipitation with Alcian blue, hyaluronan (HA) by an ELISA-like assay using biotinylated HA-binding complexes, matrix metalloproteinase 3 (MMP-3), and tissue inhibitor of metalloproteinases 1 (TIMP-1) by sandwich ELISA, and synovitis was scored by light microscopy.RESULTS: Synovitis or effusion was absent in all experimental and control groups. Immobilisation decreased the joint lavage fluid levels of IL1 (p<0.05), TIMP (p< 0.05), and the concentration of CS down to 38 (p<0.05) in comparison with untreated litter mates with normal weight bearing. Immobilisation did not affect the activity of PLA2, or the concentration of MMP-3 or HA in synovial fluid. Joint remobilisation restored the decreased concentrations of markers to control levels. Increased weight bearing did not change the concentrations of markers in comparison with the control joints with normal weight bearing.CONCLUSIONS: 11 weeks joint immobilisation decreased the concentration of markers of cartilage and synovium metabolism in the synovial fluid, and remobilisation restored the concentrations to control levels. The changes in joint metabolism induced by immobilisation, as reflected by the markers, are thus different from those found in osteoarthritis, where increased levels of these markers are associated with enhanced degradation and synthesis. These findings suggest that the change induced in joint metabolism by immobilisation is reversible in its early stages

Evolution within a language: environmental differences contribute to divergence of dialect groups

Background: The processes leading to the diversity of over 7000 present-day languages have been the subject of scholarly interest for centuries. Several factors have been suggested to contribute to the spatial segregation of speaker populations and the subsequent linguistic divergence. However, their formal testing and the quantification of their relative roles is still missing. We focussed here on the early stages of the linguistic divergence process, that is, the divergence of dialects, with a special focus on the ecological settings of the speaker populations. We adopted conceptual and statistical approaches from biological microevolution and parallelled intra-lingual variation with genetic variation within a species. We modelled the roles of geographical distance, differences in environmental and cultural conditions and in administrative history on linguistic divergence at two different levels: between municipal dialects (cf. in biology, between individuals) and between dialect groups (cf. in biology, between populations).Results: We found that geographical distance and administrative history were important in separating municipal dialects. However, environmental and cultural differences contributed markedly to the divergence of dialect groups. In biology, increase in genetic differences between populations together with environmental differences may suggest genetic differentiation of populations through adaptation to the local environment. However, our interpretation of this result is not that language itself adapts to the environment Instead, it is based on Homo sapiens being affected by its environment, and its capability to adapt culturally to various environmental conditions. The differences in cultural adaptations arising from environmental heterogeneity could have acted as nonphysical barriers and limited the contacts and communication between groups. As a result, linguistic differentiation may emerge over time in those speaker populations which are, at least partially, separated.Conclusions: Given that the dialects of isolated speaker populations may eventually evolve into different languages, our result suggests that cultural adaptation to local environment and the associated isolation of speaker populations have contributed to the emergence of the global patterns of linguistic diversity