Mechanism of kidney stone formation in chronic hemodialysis patients.
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Biomedical subjects
Publications and source records attributed to H Ozasa.
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We previously reported the isolation of a cDNA encoding the liver-specific isozyme of rat S-adenosylmethionine synthetase from a lambda gt11 rat liver cDNA library. Using this cDNA as a probe, we have isolated and sequenced cDNA clones for the rat kidney S-adenosylmethionine synthetase (extrahepatic isoenzyme) from a lambda gt11 rat kidney cDNA library. The complete coding sequence of this enzyme mRNA was obtained from two overlapping cDNA clones. The amino acid sequence deduced from the cDNAs indicates that this enzyme contains 395 amino acids and has a molecular mass of 43,715 Da. The predicted amino acid sequence of this protein shares 85% similarity with that of rat liver S-adenosylmethionine synthetase. This result suggests that kidney and liver isoenzymes may have originated from a common ancestral gene. In addition, comparison of known S-adenosylmethionine synthetase sequences among different species also shows that these proteins have a high degree of similarity. The distribution of kidney- and liver-type S-adenosylmethionine synthetase mRNAs in kidney, liver, brain, and testis were examined by RNA blot hybridization analysis with probes specific for the respective mRNAs. A 3.4-kilobase (kb) mRNA species hybridizable with a probe for kidney S-adenosylmethionine synthetase was found in all tissues examined except for liver, while a 3.4-kb mRNA species hybridizable with a probe for liver S-adenosylmethionine synthetase was only present in the liver. The 3.4-kb kidney-type isozyme mRNA showed the same molecular size as the liver-type isozyme mRNA. Thus, kidney- and liver-type S-adenosylmethionine synthetase isozyme mRNAs were expressed in various tissues with different tissue specificities.
To evaluate the usefulness of the plasma dehydroepiandrosterone (DHEA)-to-cortisol ratio (D/C) and the plasma aldosterone-to-plasma renin activity ratio (ALDO/PRA) as indicators of stress, we first monitored changes in these ratios associated with surgery in 13 patients who were healthy except for their localized gynecologic diseases. D/C and ALDO/PRA ratios were reduced by 37 and 42%, respectively, 4-5 days postsurgery compared to those 3-4 days before surgery (P less than 0.05 and P less than 0.01, respectively) and returned to preoperative levels 11-13 days after surgery. In contrast, individual hormone levels showed no significant changes associated with surgery. Having documented that these ratios may serve as indicators of stress, we then sequentially measured D/C ratios in patients with gynecologic malignancy subjected to cytotoxic chemotherapy or radiation therapy and in patients in the terminal stage. Although such therapies did not affect D/C ratios to a measurable extent, patients in the terminal stage gave consistently low D/C ratios in spite of normal vital signs (a D/C ratio below 6 was deemed low). Such low ratios occurred only sporadically in other patients and, again, individual values for DHEA and cortisol showed no consistent pattern. We believe that use of D/C ratios as an indicator of stress warrants further investigation.
We analyzed the relations between blood pressure in sitting and supine positions, left ventricular mass (LVM) and Doppler aortic, pulmonary and mitral flow velocity measurements in 163 healthy school children. Systolic blood pressure in a supine position correlated significantly with aortic acceleration (ATc) and ejection time (ETc), corrected by the square of R-R interval, pulmonary AT and peak flow velocities. Moreover, the systolic blood pressure in a sitting position correlated with pulmonary AT and LVM. LVM correlated with pulmonary ATc, the ratio of AT to ET and average acceleration, and aortic ETc and peak flow velocity. These data suggest the following: (1) the posture influences the relation between blood pressure and flow velocity, (2) pulmonary hemodynamics are influenced by systemic blood pressure in healthy children and (3) the development of LVM may be dependent not only on blood pressure but also on cardiac work in childhood.
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We have isolated cDNA clones encoding the rat liver S-adenosylmethionine synthetase by means of immunological screening from a phage lambda gt 11 expression library containing cDNA synthesized from adult rat liver poly(A)-RNA. The amino acid sequence deduced from the cDNA indicates that the rat liver enzyme for this protein contains 397 amino acid residues and has a molecular mass of 43697 Da. The deduced amino acid sequence of rat liver S-adenosylmethionine synthetase was 68% similar to those of yeast S-adenosylmethionine synthetases encoded by two unlinked genes SAM1 and SAM2. The rat liver S-adenosylmethionine synthetase also shows 52% similarity with the deduced amino acid sequence of the MetK gene encoding the S-adenosylmethionine synthetase in Escherichia coli.
cDNAs encoding the entire coding regions of the precursors (p) of rat long chain acyl-CoA (LCAD), short chain acyl-CoA (SCAD) and isovaleryl-CoA dehydrogenase (IVD) have been cloned and sequenced. Three cDNAs for rat liver LCAD together cover a 1440-base pair region. These cDNAs encode the entire 430-amino acid sequence of pLCAD, including the 30-amino acid leader peptide and the 400-amino acid mature LCAD. A single 1773 base pair cDNA for rat SCAD covers the entire coding region (414 amino acids), including the 26-amino acid leader peptide and the 388-amino acid mature peptide. Four identified IVD cDNAs, when combined, encompass a 2104 base region, and encode 424 amino acids including a 30-amino acid leader peptide and the 394-amino acid mature peptide. The identities of all cDNA clones have been confirmed by matching the amino acid sequences predicted from the respective cDNAs to the amino-terminal and tryptic peptide sequences derived from the corresponding purified rat enzyme. Comparison of the sequences of four rat acyl-CoA dehydrogenases, including LCAD, MCAD, SCAD, and IVD, and two of their human counterparts (MCAD and SCAD) reveals a high degree of homology (57 invariant and 92 near invariant residues: 30.6-35.4% of identical residues in pairwise comparisons), suggesting that these enzymes belong to a gene family and have evolved from a common ancestral gene.
Our previous report demonstrated that two human cardiac alpha- and beta-myosin heavy-chains (MHCs) which correspond to MYH6 and MYH7 respectively, according to Human Gene Mapping nomenclature, were mapped to human chromosome 14 and that human cardiac and skeletal MHC genes do not cosegregate. For further analysis, the regional mapping method was used. DNA from 4 human deletion and 3 human duplication cell lines were prepared for southern blotting, hybridized with human cardiac alpha- and beta-MHC DNA probes, and the hybridization intensity relative to 46,XX or 46,XY DNA was estimated. The results showed that two human cardiac MHC genes segregated with the 14cen----q13 region of the long arm of human chromosome 14. In situ hybridization of 3H-labeled human cardiac alpha-MHC probe to normal human metaphase chromosome independently confirmed this result.
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In a patient with mos 45,X/46,XYnf, the diagnosis was confirmed with a Y chromosome-specific DNA probe, Y-190. The patient was a phenotypic female without Turner syndrome stigmata other than short stature. She showed some evidence of virilization and high serum testosterone. Her peripheral blood karyotype was mos 45,X/46X, +mar. Although this marker chromosome resembled a Y chromosome, there was no quinacrine bright region on its long arm. Southern blot analysis of her peripheral blood mononuclear cell DNA with Y-190 as a probe showed strong hybridization with this probe. Gonadectomy was performed, and bilateral gonadoblastomas were found.
Demonstration and staging of carcinoma of the cervix with magnetic resonance (MR) imaging was evaluated prospectively in 67 patients with histologically proven lesions. Findings were correlated with surgical/pathologic results obtained within 2 weeks. MR imaging had an accuracy of 95% in demonstrating invasive disease (stage IB or higher). It was capable of depicting the location and extent of tumor invasion of cervical stroma and helped detect tumor beneath relatively normal epithelium or within the endocervical canal that had not been detected by means of colposcopic biopsy. The overall accuracy of MR imaging in staging carcinoma of the cervix was 76%, and in demonstrating parametrial status, the overall accuracy was 89%. In 39 patients with proven invasive disease, the accuracy in demonstrating parametrial status was 82%. In 13 of these 39 patients the low-signal-intensity stromal ring of the cervix on MR images was completely preserved and there were no false-positive results. MR imaging is a highly promising method for directly demonstrating and staging carcinoma of the cervix and seems to be capable of providing answers to crucial questions regarding mode of therapy.
The potential of magnetic resonance (MR) imaging in differentiation of adenomyosis from leiomyoma was evaluated in 93 patients who had a palpable enlarged uterus that was suspect for leiomyoma or adenomyosis. In all cases, MR images were correlated with surgical/pathologic findings. Pathologic findings showed that 71 enlarged uteri were due to leiomyoma, including one leiomyosarcoma, and 16 were due to adenomyosis. The other six patients were shown to have an enlarged uterus attributable to simultaneous involvement of both lesions. On T2-weighted images, adenomyosis appeared as an ill-defined, relatively homogeneous low-signal-intensity area embedded with sparse high-intensity spots. In contrast, leiomyomas were well-circumscribed masses with a spectrum of signal intensity. The cause of uterine enlargement was correctly diagnosed with MR images in 92 of the 93 cases. It is concluded that MR imaging is highly accurate in helping to distinguish between adenomyosis and leiomyoma in cases of enlarged uterus.
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We have examined amyloid-like kidney stones, commonly found in patients on maintenance hemodialysis. Extraction of protein from the stones and Western blot analysis were performed. beta 2-Microglobulin (beta 2MG), serum amyloid P component (SAP), lysozyme and PAS-positive substance were identified in the stones. It is suggested that calcium-mediated association of beta 2MG, lysozyme, SAP and PAS-positive substance may have an important role in the process of the formation of kidney stones in chronic hemodialysis patients.
Complementary DNAs encoding the precursor of human placental short chain acyl-coenzyme A (CoA) dehydrogenase (SCAD) (EC 1.3.99.2) were cloned and sequenced. The cDNA inserts in these clones were 1,852 bases in length combined, and encoded the entire 412-amino acid precursor SCAD (mol wt 44,303). This sequence included the 24-amino acid leader peptide moiety (mol wt 2,576) and 388 amino acids corresponding to the mature protein (mol wt 41,727). The comparison of SCAD and medium chain acyl-CoA dehydrogenase sequences revealed a high degree of homology, suggesting that these enzymes evolved from a common ancestral gene and belong to a gene family. We also studied mutant human SCAD in cultured skin fibroblasts from three patients with hereditary SCAD deficiency. Labeling fibroblast cultures with [35S]-methionine followed by immunoprecipitation with anti-SCAD antibody revealed that a normal size variant SCAD protein was synthesized. In all of the three SCAD-deficient cell lines, the size of variant SCAD mRNA as determined by Northern blotting using one of the normal SCAD cDNA as a probe was also normal, and no difference was observed on Southern blots in the restriction patterns of mutant genomic DNA using EcoRI, TaqI, HincII, and BamHI. These results suggest that the defects in SCAD in these cell lines are caused by a point mutation.