5 research outputs found
Prolyl 4-hydroxylase:genomic cloning of the human and mouse α(II) subunit, tissue distribution of type I and II isoenzymes, and cloning and characterization of a novel prolyl 4-hydroxylase from Caenorhabditis elegans
Abstract
The collagens are a family of extracellular matrix proteins with a
widespread tissue distribution. Collagen biosynthesis requires the hydroxylation
of a number of proline residues by prolyl 4-hydroxylase. This posttranslational
modification is essential for the synthesis of all collagens, as 4-hydroxyproline
deficient collagens cannot form stable triple helices at body temperature.
The genes for the human and mouse prolyl 4-hydroxylase α(II) subunits
were cloned and characterized in this study. The human and mouse genes are 34.6
and 30.3 kb in size, respectively, consisting of 16 exons and 15 introns. The
intron sizes vary from 48-49 bp to over 8 kb in both genes. The 5' flanking
regions contain no TATA box, but there are several motifs that may act as
transcription factor binding sites. A novel mutually exclusively spliced exon 12a
was identified in both genes. Both variants of the α(II) subunit were found
to be expressed in a variety of tissues and both formed a fully active
recombinant tetramer with the β subunit when expressed in insect
cells.
Tissue distribution of the type I and type II prolyl 4-hydroxylase
isoenzymes was studied in developing, mature, and malignant cells and tissues by
immunofluorescence and Western blotting. The results indicate that the type I
isoenzyme is the main form in many cell types. Skeletal myocytes and smooth
muscle cells appeared to have the type I isoenzyme as their only prolyl
4-hydroxylase form, whereas the type II isoenzyme was clearly the main form in
chondrocytes. A strong signal for the type II enzyme was detected in cultured
umbilical and capillary endothelial cells, whereas the type I isoenzyme could not
be detected in these cells by immunostaining or Western blotting. Similar studies
on primary chondro- and osteosarcomas and benign bone tumours indicated that the
type I isoenzyme is the predominant form in both types of bone sarcoma, whereas
the type II isoenzyme was more abundantly expressed in benign tumours. In
chondrosarcomas, the type II isoenzyme was expressed in the nonmalignant
chondrocytes, whereas their malignant counterparts switched their expression
pattern to that of the type I isoenzyme.
Two isoforms of the catalytic prolyl 4-hydroxylase α subunit, PHY-1
and PHY-2, have previously been characterized from Caenorhabditis
elegans. This study reports the cloning and characterization of a
third C. elegans α subunit isoform, PHY-3, which is
much shorter than the previously characterized vertebrate and C.
elegans α subunits. Nematodes homozygous for a
phy-3 deletion were phenotypically wild type and fertile,
but the 4-hydroxyproline content of their early embryos was reduced by about 90%.
The expression of PHY-3 was found to be restricted to spermatheca of late larvae
and adult nematode, indicating that PHY-3 is likely to be involved in the
synthesis of collagens of the early embryo egg shells
Histological findings of patients with adnexal torsion who underwent surgical treatment:short reminder
Abstract
Background: Ovarian torsion is a rare emergency condition in women. Early diagnosis is necessary to preserve fertility.
Case: Our study evaluated 40 patients, who underwent laparoscopic surgery. The aims of this retrospective study were to emphasize the importance of early diagnosis in ovarian torsion, evaluate the process of patient treatment, and investigate the number of patients treated by minimally invasive surgery. In this article, we present the outcomes from the patient data.
Results: Thirty-two percent (13/40) of patients were first evaluated by the surgeon to investigate right-sided lower abdominal pain. These patients were first misdiagnosed with appendicitis or urinary tract stones. Among these patients, necrotic ovary tissue was more common, most likely due to a longer delay seeking medical attention. A total of 77% (31/40) of patients underwent laparoscopic surgery on the same date that they were admitted to the hospital. No severe complications occurred in this group of patients. All histological findings were benign. In 52% (21/40) of patients, the adnexa was removed, whereas in 37% (15/40) of patients the torsed adnexa was treated by detorsion. A total of 27% (11/40) of patients had no diagnosis before undergoing surgery.
Conclusions: Rapid and accurate diagnosis is essential to preserve ovarian function
The serum levels of circulating matrix metalloproteinase MMP-9, MMP-2/TIMP-2 complex and TIMP-1 do not change significantly during normal pregnancy:a pilot study
Abstract
Objective: Matrix metalloproteinases (MMPs) are important regulators of vascular and uterine remodeling. They exhibit proteolytic activity implicating the efficiency of trophoblast invasion to the uterine wall involving marked hemodynamic and uterine changes. In this pilot study sera of 13 women with normal pregnancy was analyzed to evaluate the usage of MMPs as diagnostic tool. The concentrations of circulating MMP-9, MMP-2/TIMP-2 complex and TIMP-1 in different time points during normal pregnancy has not been studied. The serum levels of MMP-9, TIMP-1, TIMP-2 and MMP-2/TIMP-2 complex were determined by enzyme-linked immunosorbent assay (ELISA). Using the same method, we have shown that serum MMPs are elevated in spontaneous early pregnancy failure as compared to normal pregnancy.
Results: The serum levels of MMP-9 and TIMP-1 were stable throughout pregnancy. The level of MMP-2/TIMP-2 complex was slightly increased after week 15 without statistical significance. For our best knowledge, this is a first study of the serum levels of MMP-9, MMP-2/TIMP-2 and TIMP-1 on different time points during normal pregnancy. Further measurements with the correlation to the outcome of the pregnancy are needed
TUFT1, a novel candidate gene for metatarsophalangeal osteoarthritis, plays a role in chondrogenesis on a calcium-related pathway
Abstract
Osteoarthritis (OA) is the most common degenerative joint disorder and genetic factors have been shown to have a significant role in its etiology. The first metatarsophalangeal joint (MTP I) is highly susceptible to development of OA due to repetitive mechanical stress during walking. We used whole exome sequencing to study genetic defect(s) predisposing to familial early-onset bilateral MTP I OA inherited in an autosomal dominant manner. A nonsynonymous single nucleotide variant rs41310883 (c.524C>T, p.Thr175Met) in TUFT1 gene was found to co-segregate perfectly with MTP I OA. The role of TUFT1 and the relevance of the identified variant in pathogenesis of MTP I OA were further assessed using functional in vitro analyses. The variant reduced TUFT1 mRNA and tuftelin protein expression in HEK293 cells. ATDC5 cells overexpressing wild type (wt) or mutant TUFT1 were cultured in calcifying conditions and chondrogenic differentiation was found to be inhibited in both cell populations, as indicated by decreased marker gene expression when compared with the empty vector control cells. Also, the formation of cartilage nodules was diminished in both TUFT1 overexpressing ATDC5 cell populations. At the end of the culturing period the calcium content of the extracellular matrix was significantly increased in cells overexpressing mutant TUFT1 compared to cells overexpressing wt TUFT1 and control cells, while the proteoglycan content was reduced. These data imply that overexpression of TUFT1 in ATDC5 inhibits chondrogenic differentiation, and the identified variant may contribute to the pathogenesis of OA by increasing calcification and reducing amount of proteoglycans in the articular cartilage extracellular matrix thus making cartilage susceptible for degeneration and osteophyte formation
Association of prevalent vaginal microbiome of mother with occurrence of type I diabetes in child
Abstract
Type I diabetes (T1D) is a rapidly increasing autoimmune disease especially in the Western countries and poses a serious global health problem. Incidence of T1D cannot be fully explained by genetic background, and environmental factors have been assumed to play a role. Environmental conditions and composition of human microbiome have been found to correlate with the incidence of T1D. We asked whether mothers’ prevalent vaginal microbiome could correlate with the incidence of T1D in child. To test this hypothesis, we collected samples of vaginal microbiomes from eight mothers that had at least one child with T1D (child age maximum of 11 years at the time of sampling), born with a vaginal delivery. Eight control mothers had child/children with vaginal delivery and no diabetic child/children. The microbiomes were studied by using 16S rRNA Ion Torrent high throughput sequencing. We found that composition of total and Lactobacillus microbiome was altered, and saw an indication that diversity of vaginal microbiomes of the mothers with a diabetic child could be higher. Based on these pilot observations, we strongly encourage a larger population study to verify whether mother vaginal microbiome diversity and composition are linked to the prevalence of T1D in children