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Biomedical subjects

H H Hsu

Publications and source records attributed to H H Hsu.

At least 55 records · Page 3Linked to original sources

Recurrent and acquired hepatitis C viral infection in liver transplant recipients.

To examine the postliver transplant recurrence of hepatitis C virus (HCV) infection in patients with pretransplant infection, as well as its acquisition in patients without prior infection, we used the polymerase chain reaction to amplify HCV RNA in serum and/or liver samples of 89 patients with alcoholic and cryptogenic cirrhosis undergoing liver transplantation. Results were correlated with histologic findings from posttransplant liver biopsies. Ninety-five percent of patients with pretransplant infection had posttransplant viremia. In contrast, 35% of patients without pretransplant infection acquired the virus (P less than 0.0001). Pretransplant HCV infection predisposed patients to hepatitis in the new graft. HCV RNA was present in serum of 96% of patients with posttransplant hepatitis. Fifty-six percent of patients with posttransplant HCV infection had no evidence of liver damage at least 1 year posttransplant. However, of those patients with histologic hepatitis, chronic active hepatitis was common. It is concluded that although HCV infection recurs posttransplant in almost all infected patients, acquisition of the HCV infection with transplant is common. Pretransplant HCV infection is an independent risk factor for the development of posttransplant hepatitis. HCV infection accounts for the majority of posttransplant hepatitis not due to cytomegalovirus, and although many patients with posttransplant viremia have little evidence of histologic hepatitis, significant hepatic damage may occur.

Base Sequence↗

Comparison of normal and rachitic rat matrix vesicles.

Accumulated information about MVs in PO4-deficient rachitic rats shows them to be normal in most aspects. Both normal and rachitic MVs show the same ultrastructural features and selective spatial distribution in the growth plate, and both contain a nearly identical array of major proteins. Rachitic and normal MVs show the same avidity to calcify in vivo and in vitro. A slightly greater specific activity of ALP in rachitic MVs may enhance their calcifiability. We conclude that rachitic rat MVs are essentially 'normal' and can be used as an adequate model to study the mechanism of biological calcification.

Alkaline Phosphatase↗

Bone-inducing agent (BIA) from cultured human Saos-2 osteosarcoma cells.

The Saos-2 line of human osteosarcoma cells was established in culture in 1975. These cells produce a large amount of alkaline phosphatase but little or no matrix in vitro, and are unable to grow when transplanted into athymic mice. We decided to test our local strain of Saos-2 cells for bone-inducing ability in the skeletal muscle of athymic mice by implanting freeze-dried, acetone-defatted cells, with and without a collagen carrier. A bone-inducing activity (BIA) thus was demonstrated in 88% of 90 implants of devitalized Saos-2 cells. In further studies, we have used guanidinium hydrochloride (Gu-HCl) to extract, solubilize, and remove the Saos bone-inducing agent(s) in an active state which when reprecipitated by aqueous dialysis was able to induce ultrastructurally typical endochondrial bone formation in nude mouse muscle in 92% of 48 implants. This preliminary report is offered to alert investigators to the presence of an extractable BIA in Saos-2 cells.

Animals↗

Neoplasms and related proliferative lesions in control Sprague-Dawley rats from carcinogenicity studies. Historical data and diagnostic considerations.

Historical data are presented for neoplasms and related proliferative lesions from 1,170 Sprague-Dawley rats that served as controls in 9 carcinogenicity (2 year) studies conducted in the Safety Evaluation Facility of Ciba-Geigy Corporation, Summit, New Jersey. The most common neoplasm was pituitary adenoma, which occurred in 62.2% of the male and 84.7% of the female rats. Incidences of other neoplasms that occurred in more than 6.0% of the rats were, for males, benign pheochromocytoma (19.0%), cutaneous keratoacanthoma (7.9%), pancreatic islet cell adenoma (7.5%), benign testicular interstitial cell tumor (6.5%), and thyroid C-cell adenoma (6.5%). For females these incidences were mammary fibroadenoma (31.3%), mammary adenocarcinoma (16.8%), and mammary adenoma (6.5%). Focal cortical hypertrophy/cystic degeneration of the adrenal, a focal nonneoplastic lesion of zona fasciculata cells that often degenerate into large cysts, was present in 23.4% of all male and 82.7% of all female rats. Criteria for the differential diagnoses of selected neoplasms and related lesions are presented.

Animals↗

Conversion of N-(2-chloroethyl)-4-piperidinyl diphenylacetate (4-DAMP mustard) to an aziridinium ion and its interaction with muscarinic receptors in various tissues.

A 2-chloroethylamine derivative [N-(2-chloroethyl)-4-piperidinyl diphenylacetate (4-DAMP mustard)] of the selective muscarinic antagonist N,N-dimethyl-4-piperidinyl diphenylacetate (4-DAMP) was synthesized, and its conversion to an aziridinium ion and interaction with muscarinic receptors was investigated. When dissolved in aqueous solution at pH 7.4 and 37 degrees, 4-DAMP mustard released an equivalent amount of chloride. The release of chloride was consistent with a first-order process having a half-time of 5.7 min. The aziridinium ion reached a peak concentration at 32 min, corresponding to 75% of the initial concentration of 4-DAMP mustard. When homogenates of rat brain, heart, and submaxillary gland were incubated with 4-DAMP mustard (9 nM) for 1 hr, washed extensively, and then assayed for muscarinic receptor binding properties, a 56% decrease in the binding capacity of N-[3H]methylscopolamine in the heart and brain and a 71% decrease in the gland were observed, without a significant change in the dissociation constants. The affinity of 4-DAMP mustard and its transformation products for muscarinic receptors was determined in competitive binding experiments with N-[3H] methylscopolamine, and the results show that the aziridinium ion of 4-DAMP mustard was the most potent form, compared with the parent 2-chloroethylamine (4-DAMP mustard) and the alcoholic hydrolysis product. The rates of receptor alkylation by 4-DAMP mustard were measured in the rat heart and gland. Virtually no alkylation (less than 1%) occurred in the heart at a 4-DAMP mustard concentration of 1.6 nM, after 30 min, whereas almost 50% alkylation was observed in the gland under the same conditions. Almost complete alkylation of receptors in the gland could be achieved at a 4-DAMP mustard concentration of 200 nM, after 1 hr. Treatment of the isolated rat ileum with 4-DAMP mustard caused an irreversible blockade of contractions elicited by the muscarinic agonist oxotremorine-M, and this blockade persisted after extensive washing. The results presented here show that 4-DAMP mustard forms an aziridinium ion that binds irreversibly to muscarinic receptors and exhibits selectivity for M3, compared with M2 muscarinic receptors.

Alkylation↗

Gastric bezoar caused by lecithin: an unusual complication of health faddism.

We discovered an unusual bezoar in a previously healthy man who had been ingesting large amounts of a vegetable-derived oil touted to contain lecithin, which he purchased from health food stores in the belief that it had beneficial effects in lowering cholesterol levels and improving memory. The large intragastric mass, composed of fatty acids and lecithin, led to considerable morbidity, including abdominal pain, early satiety, and significant weight loss, and required surgical removal.

Bezoars↗

Failure to detect hepatitis C virus genome in human secretions with the polymerase chain reaction.

Although hepatitis C infection has been clearly demonstrated to be transmitted through blood products or blood contamination, most cases of sporadic hepatitis C infection are unassociated with parenteral risk factors, and it is unclear how infection might be acquired by nonparenteral means. One potential mode of nonparenteral transmission is through body secretions. We used a highly sensitive and specific polymerase chain reaction assay to determine whether hepatitis C viral genomic RNA could be detected in secretions obtained from nineteen individuals with chronic hepatitis C virus infection. Although hepatitis C genomic RNA was found in all 19 sera, hepatitis C virus RNA was not detected in any samples of saliva, semen, urine, stool or vaginal secretions from these patients. Viral titers in serum ranged from 10(2) to 10(7) polymerase chain reaction units/ml. The sensitivity of our polymerase chain reaction assay indicates that, if hepatitis C virus were in secretions, it would be present in amounts less than 1 to 4 polymerase chain reaction units/ml. This contrasts with hepatitis B virus infection, in which serum titers frequently are in excess of 10(9) copies of hepatitis B genomes/ml. Body secretions have been found to contain up to 10(6) copies of hepatitis B genomes/ml. Our findings support seroepidemiological studies indicating that nonparenteral transmission of hepatitis C through secretions is uncommon and probably much less efficient than hepatitis B virus infection.

Body Fluids↗

Antibodies to hepatitis C virus in low-risk blood donors: implications for counseling positive donors.

The significance of antibodies to hepatitis C virus (HCV) found by screening enzyme-linked immunoassay testing in a low-risk blood donor population is unclear. The rate of false positivity in this group as well as the usefulness of supplemental testing were examined by correlating the results of two screening enzyme-linked immunoassays (Ortho Diagnostics, Raritan, NJ, and Abbott Laboratories, North Chicago, IL) with supplemental antibody testing by the recombinant immunoblot assays (1 and 2) (Ortho) and neutralization assay (Abbott). Polymerase chain reaction was used to detect HCV genomic RNA to confirm viremia. Among 11.243 volunteer donors who were screened for the presence of antibodies to HCV by enzyme-linked immunoassay, 60 (0.53%) sera were repeatedly reactive. Twenty-five of these 60 sera were available for further testing. Seven sera were reactive by both screening enzyme-linked immunoassays, as well as by both recombinant immunoblot and neutralization assays. Six of these seven sera had detectable HCV genomic RNA by polymerase chain reaction. Among the remaining 18 sera, none were reactive by either recombinant immunoblot assays, whereas two sera were reactive by the neutralization assay only. None of the 18 samples had detectable HCV genomic RNA. Five of the six sera with elevated aminotransferase levels were among the seven sera reactive by all immunoassays. It is concluded that there is a significant false positivity rate associated with screening enzyme-linked immunoassay testing in a low-risk blood donor population. Supplemental testing correlates well with detection of hepatitis C genomic material by polymerase chain reaction and identifies donors who are truly infected.

Base Sequence↗

The partial sequencing of human nonspecific (bone/liver/kidney) alkaline phosphatase gene.

1. A charon 4A human fetal liver genomic library was screened for human nonspecific alkaline phosphatase gene using the cloned human bone cDNA as a hybridization probe. 2. A clone 2.2 Kb DNA was sequenced and found to contain a piece of sequences encoding the 4-44th amino acids of NH2 terminus. 3. The other cloned 1.6 Kb DNA contains two segments of sequences each corresponding to two separate regions of the cDNA for alkaline phosphatase. The first segment of the DNA codes for the 83-141st amino acids whereas the second for 141-199th.

Alkaline Phosphatase↗

Delay of diagnosis of growth hormone deficiency in Taiwan.

In view of the importance of early detection of growth hormone deficiency, the height and age at the first pediatric endocrinologic clinic visit of 45 patients with growth hormone (GH) deficiency between January 1988 to December 1989 were retrospectively analyzed. Twenty-three patients with idiopathic GH deficiency had a median age of 13 years (range, from 3.8 to 29.9 years) at their first visit and a median height standard deviation score (SDS) of -3.87 (-2.14 to -7.93). Nineteen children with hypothalamopituitary tumors at their first visit, at a median of 2.17 years (0.42 to 7.83 years) after diagnosis of the tumor, had a median height SDS of -3.15 (-0.58 to -4.70). Three children with GH deficiency secondary to cranial irradiation at their first visit, at 8.17 years (1.33 to 10 years) after treatment for their malignancy, had a height SDS of -3.15 (-2.13 to -3.72). It is well known that the earlier the replacement therapy is begun in such patients, the better the prognosis is for final adult height. Regular height measurement and routine screening for shortness at school entry may overcome this delay in diagnosis. In addition, patients who have received cranial irradiation or had hypothalamopituitary tumors should be regularly measured and studied if their height velocities become subnormal. Physicians dealing with children should keep the possibility of growth hormone deficiency in mind when examining a short child.

Adolescent↗

Matrix vesicle biogenesis in vitro by rachitic and normal rat chondrocytes.

Calcifying matrix vesicles (MVs) are released from chondrocytes and osteoblasts in monolayer culture. In the present studies, we tested the ability of rachitic versus normal rat growth plate chondrocytes in micromass or monolayer primary cultures to produce MVs. Unlike earlier reports of in vitro MV biogenesis by chicken chondrocytes in which most MVs were released into the medium, we found that most of the released rat matrix vesicles were entrapped in a newly formed cartilaginous matrix enveloping the cells. These matrix-associated MVs could be isolated by mild collagenase treatment and concentrated by differential centrifugation. Vesicle production slowed in the older 2- to 4-week-old cultures and, unlike vesicle release from cultured chicken chondrocytes, active vesicle production did not show a second burst of activity at 3 to 4 weeks. Alkaline phosphatase (ALP) activity diminished with time in culture in cells and matrix vesicles, suggesting a decrease in differentiative expression. Protein profiles on SDS polyacrylamide gels of native matrix vesicles and culture-derived MVs from rachitic and normal cells were quite similar and showed a typical simplified protein pattern as compared to chondrocyte plasma membrane proteins. There were distinctive proteins migrating at 130, 80 to 95, 66, 43, 20, and 14 kd. Culture-derived MVs showed vigorous in vitro calcifying activity that was ALP related. We conclude that 1) rachitic chondrocytes are essentially normal in their matrix vesicle production; 2) matrix entrapment of MVs is a characteristic of rat chondrocyte cultures; and 3) culture-produced MVs are similar to native MVs in protein profile and calcifiability, and thus can be studied as a model for normal MV composition and calcification.

Animals↗

The isolation and partial sequencing of human bone alkaline phosphatase gene.

1. A charon 4A human fetal liver genomic library was screened for human alkaline phosphatase sequences using the cloned human bone cDNA as a hybridization probe. 2. A positive clone was obtained and then characterized by restriction endonuclease cleavage analysis, hybridization experiments and partial DNA sequencing.

Alkaline Phosphatase↗

Purification and partial amino acid sequencing of alkaline phosphatase from rachitic rat epiphyseal cartilage.

1. Alkaline phosphatase of rachitic epiphyseal cartilage was purified to apparent homogeneity by sequential application of monoclonal affinity, DEAE-cellulose, and Sepharose CL-6B chromatography. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the enzyme showed the presence of a dominant band corresponding to a molecular weight of 80,000. 2. The N-terminal amino acid sequence was determined as follows: Phe-Val-Pro-Glu-Lys5-Glu-Lys-Asp-Pro-Ser10-Tyr-Trp-Arg-Gln-+ ++Gln15-Ala-Gln-Glu- Thr-Leu20-Lys-Asn-Ala-Leu-Lys25-Leu-Gln-Lys-?-Asn-Val-Asn-?- Ala-Lys35-?-Ile-?- Met-Phe40-Leu-(Gly?)-Asp-(Ala/Gly?)-Met45-?-Val-?- (Val/Gly?).

Alkaline Phosphatase↗

Purification and partial amino acid sequencing of rat bone tumor (UMR106) alkaline phosphatase.

Cultured rat osteosarcoma (UMR106) alkaline phosphatase was purified to apparent homogeneity by sequential application of polyclonal antibody affinity, DEAE-cellulose, and Sepharose CL-6B chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the enzyme preparation treated with sodium dodecyl sulfate and mercaptoethanol showed the presence of a dominant band (using silver staining) corresponding to a molecular weight of 80,000. The amino acid composition was similar to those of various alkaline phosphatases. The N-terminal amino acid sequence was determined as follows: Phe-Val-Pro-Glu-Lys-Glu-Lys- Asp-Pro-Ser-Tyr-Trp-Arg-Gln-Gln-Ala-Gln-Glu-Thr-Leu- Lys-Asn-Ala-Leu-Lys-?-Gln-Lys-?-Asn-Val-Asn-Ala-Lys.

Alkaline Phosphatase↗

Purification and partial amino acid sequencing of bovine kidney alkaline phosphatase.

Bovine kidney alkaline phosphatase (ALPase) was purified by the sequential application of monoclonal anti-bovine cartilage ALPase affinity, DEAE-cellulose, and Sepharose CL-6B chromatography. Sodium dodecyl sulfate-polyacrylamide-gel electrophoresis showed the presence of a single band corresponding to a molecular weight of 80,000. The N-terminal amino acid sequence of bovine kidney alkaline phosphatase was determined as follows: Leu-Val-Pro-Glu-Lys-Asp-Pro-?-Tyr-Trp-Arg-Asp-Gln-Ala-Gln.

Alkaline Phosphatase↗

Purification of bone alkaline phosphatase from human osteosarcoma.

Purification of human bone alkaline phosphatase, derived from human osteosarcoma tissue, has been carried to electrophoretic homogeneity. The purification procedure involved three major steps: (1) chromatography on hydroxylapatite; (2) ion exchange chromatography; and (3) gel filtration. The resultant purified enzyme is a glycoprotein, has a molecular weight of approximately 80,000 (consistent with previous reports for the bone isoenzyme), and is characteristically inhibited by modest heat (56 degrees C, 30 min) and L-homoarginine but not by L-phenylalanine. The isolation and purification procedure described can lead to the production of significant amounts of highly purified bone alkaline phosphatase. Purified ALP can be used for an analysis of minor structural differences that appear to exist between the bone, liver and kidney isoenzymes. Such information could lead to the development of a clinical diagnostic procedure specifically for bone alkaline phosphatase.

Alkaline Phosphatase↗