Pitfall in the implication of intercellular adhesion molecule-1 expression on isolated hepatocytes.
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Animals↗
Biomedical subjects
K Fujiwara
Publications and source records attributed to K Fujiwara.
Up-regulation of type I procollagen C-proteinase enhancer protein messenger RNA in rats with CCl4-induced liver fibrosis.
Using a polyclonal antibody raised against a liver stellate cell (LSC) line derived from a rat CCl4-cirrhotic liver, we isolated 14 clones from a complementary DNA library prepared with total RNA extracted from the same cell line, with nucleotide sequences homologous to that of the type I procollagen C-proteinase enhancer protein (PCPE) gene. The longest PCPE insert of 1,530 base pairs contained an open reading frame coding for 468 amino acids. PCPE cDNA recognized by Northern blot a 1.7-kilobase messenger RNA (mRNA) in total RNA extracted from freshly isolated and early passaged LSC, LSC lines derived from normal (NFSC) and cirrhotic (CFSC) rat livers, and various LSC clones derived from CFSC. The expression of PCPE mRNA was increased threefold in CFSC compared with NFSC. PCPE mRNA was not detected in total rat liver, freshly isolated hepatocytes, or endothelial or Kupffer cells. However, the expression of PCPE mRNA was induced in fibrotic livers of rats treated with CCl4. PCPE mRNA expression in LSC was up-regulated by transforming growth factor beta1 (TGF-beta1) and down-regulated by tumor necrosis factor alpha (TNF-alpha), similar to the changes in alpha1 (1) procollagen mRNA induced by these cytokines. PCPE was not detectable in liver biomatrix proteins obtained from normal liver. However, PCPE was increased in liver biomatrix proteins from cirrhotic livers and was proportional to the amount of collagen. These data suggest that PCPE may play an important role in the processing of type I collagen during liver fibrogenesis, and that TGF-beta1 and TNF-alpha regulate its expression.
Amino Acid Sequence↗
Transforming growth factor-alpha (TGF alpha)-producing gastric carcinoma with acanthosis nigricans: an endocrine effect of TGF alpha in the pathogenesis of cutaneous paraneoplastic syndrome and epithelial hyperplasia of the esophagus.
A case of well-differentiated adenocarcinoma (Borrmann type 3) of the stomach in a 76-year-old man associated with the typical skin manifestations of acanthosis nigricans and with multiple protruding lesions showing epithelial hyperplasia of the esophagus is reported. The advanced tumor was located in the cardiac region of the stomach, and measured approximately 8 cm in diameter, with partial invasion to the esophagus. The associated cutaneous lesions were characterized by hyperpigmentation and by protruding verrucous papules on the torso, head, face, neck, upper extremities, perineum, and inguinal region. Histologically, the protruding skin lesions showed keratinocytes proliferation throughout the epidermis, resulting in diffuse hyperkeratosis, papillomatosis, and acanthosis of the skin. Immunohistological analysis showed coexpression of transforming growth factor alpha (TGF-alpha) and epidermal growth factor (EGF) receptors in the tumor from the stomach. It is reasonable to conclude from this evidence that gastric carcinoma cells secrete TGF alpha in an autocrine for auto-stimulation. EGF receptor expression was also noted on the papillomatous hyperplasia of the cutaneous lesion. Serum level of TGF alpha, determined by an enzyme-linked immunosorbent assay, was high (144 pg/ml; normal, 22.0 +/- 16 pg/ml (Mean +/- SD)). Serum TGF alpha abruptly decreased to 49 pg/ml on day 7 after the total gastrectomy, and then gradually increased to 77 pg/ml within 28 days. Amelioration of the cutaneous lesions and the protruding lesions in the esophagus was observed after surgical resection of the gastric carcinoma. This suggests that the TGF alpha stimulates the proliferation of keratinocytes involved with EGF receptor. Large amounts of circulating TGF alpha in the blood over a long period released by the primary tumor seem to act as an endocrine-like mechanism causing epidermal and esophageal epithelial cells to proliferate. There is a possible link in the pathogenesis of the acanthosis nigricans as a cutaneous paraneoplastic syndrome, and epithelial hyperplasia of the esophagus.
Acanthosis Nigricans↗
Monoclonal antibodies against glutaraldehyde-conjugated histamine: application to immunocytochemistry.
We have developed mouse monoclonal antibodies (AHA-1-5, all IgG1 sub-isotype mAbs) against histamine (HA) conjugated to bovine serum albumin using glutaraldehyde-NaBH4. Among these, AHA-1 mAb was found to be the most useful for HA immunocytochemistry (ICC) in terms of specificity and sensitivity without non-specific immunobinding. AHA-1 was demonstrated to be specific to HA with an enzyme-linked immunosorbent assay (ELISA) binding test, simulating the ICC of tissue sections, and not reactive to any of the other amino acids and peptides with N-terminal histidine tested. By use of this antibody, indirect immunoperoxidase staining was observed in rat stomach fixed with glutaraldehyde (GA) in combination with NaBH4 reduction. In contrast, no immunoreactivity was seen in tissue fixed only with GA. Absorption controls indicated that the immunostaining could be completely inhibited by GA-conjugated HA, which was consistent with the results of an ELISA inhibition test. No cross-reactivity occurred with other GA-conjugated amino acids. ICC staining was dense in the cytoplasm of gastric enterochromaffin-like cells and very weak in mast cells. A new finding was that staining was noticed in some cell bands of the intermediate layer between the stratum lucidum and the stratum corneum of the stratified squamous epithelium of the gastric cardia, esophagus, tongue, and skin in rats. The results strongly suggest that the monoclonal antibody allowed highly specific detection of HA in animal tissues.
Animals↗
Surgical strategy for doubly committed subarterial ventricular septal defect with aortic cusp prolapse.
BACKGROUND: Many surgeons recommend early repair of doubly committed subarterial ventricular septal defect regardless of the clinical symptoms. We reviewed our patients of this anomaly with aortic cusp prolapse to justify this strategy. METHODS: We reviewed the preoperative and postoperative records of 27 patients with doubly committed subarterial ventricular septal defect and aortic cusp prolapse. The patients' ages ranged from 2 months to 11 years (median, 4.6 years). RESULTS: During the preoperative observation period, aortic regurgitation (AR) developed in 65% of the patients. In the 8 patients without AR before the operation, AR did not develop after the operation, whereas AR persisted in 12 (63%) of the 19 patients with preoperative AR. To identify the risk factors for persistent AR after the operation we analyzed the data for the patients with preoperative AR in the persistent AR group (n = 12) and eliminated AR group (n = 7) and found a longer period from the onset of AR to the operation in the persistent AR group (32.1 +/- 10.1 versus 5.6 +/- 1.9 months; p = 0.014). During the follow-up period 10 of the 17 patients with mild AR before the operation showed persistent AR in the postoperative period, but it did not progress. CONCLUSIONS: We conclude that early surgical repair with a minimum observation period is essential for prevention of residual AR. Even if a tiny AR is detected preoperatively, the patient should be surgically treated immediately.
Age Factors↗
Subcutaneous transposition of the ovary following hysterectomy.
OBJECTIVES: Preservation of ovarian function at hysterectomy in pre-menopausal women is desirable. A possibly malignant tumor from retained ovary is a concern. We demonstrate a new technique for exteriorization of the ovaries. METHOD: In 27 patients, the infundibulopelvic ligament was mobilized. The retroperitoneal space was then widely opened. The ovary was gently pulled through a fascial incision and fixed on the fascia followed by skin closure. RESULTS: Cystic formation was common (66.7%). Four patients had extreme enlargement of the transposed ovary. A needle puncture revealed intra-ovarian hematomas or fluid retention. One had puncture of the translocated ovary for in vitro fertilization more than two years following transposition. CONCLUSIONS: Possible advantages are (1) early detection and diagnosis of ovarian cysts, (2) easy surgical access to remove ovarian cysts, (3) transposition of ovaries outside the radiation field and (4) in vitro fertilization is facilitated. Comparative study for ovarian function is ongoing.
Abdomen↗
Effects of neurotropic pyrimidine heterocyclic compound, MS-430, on cultured hepatic parenchymal and stellate cells.
MS-430 is a novel synthetic pyrimidine derivative that stimulates regeneration of the nerve as a promoter for various growth factors such as epidermal growth factor (EGF) and nerve growth factor, and differentiation of astrocytes. The effects of MS-430 on the liver were tested using hepatocytes and stellate cells in primary culture isolated from rats. MS-430 enhanced EGF-induced DNA synthesis in hepatocytes while it alone failed to increase the basal DNA synthesis. Albumin mRNA expression in the cells and its amount in the medium were not changed by addition of EGF or MS-430 alone or both. Basic fibroblast growth factor (bFGF) increased DNA and but not collagen synthesis by hepatic stellate cells. Addition of MS-430 inhibited DNA synthesis by hepatic stellate cells at either presence or absence of bFGF, and collagen synthesis at the presence of bFGF. However, MS-430 had no effects on basal or bFGF-stimulated TGFbeta mRNA expression in the cells. These results suggest that MS-430 stimulated proliferation of hepatocytes as a comitogen for EGF without affecting albumin synthesis, and suppressed proliferation of activated hepatic stellate cells and their collagen synthesis without affecting TGFbeta expression.
Animals↗
Lipoate addition to acyltransferases of alpha-keto acid dehydrogenase complexes and H-protein of glycine cleavage system.
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3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗
Relationship between radical production and natural killer cytotoxic factor (NKCF) in canine natural killer (NK) cell-mediated cytotoxicity.
The relationship between radical production and natural killer cytotoxic factor (NKCF) release via canine natural killer (NK)-mediated cytotoxic mechanism was examined. Radical production and NKCF release was induced in NK cells stimulated with either dead target cells, or their cytoplasmic membranes, as well as live target cells. Canine NKCF evoked target cell lysis but did not induce radical production. Radical production was inhibited by the addition of Tiron or n-propyl gallate, whereas NK-mediated cytotoxicity and NKCF release were only inhibited by the addition of n-propyl gallate. These results suggested that radical production and NKCF release may be induced by the contact and binding of NK cells to the target cell cytoplasmic membrane. Therefore, the release of NKCF from NK cells attached to the target cell cytoplasmic membrane may be associated with the production of radicals, especially hydroxyl radicals.
Animals↗
Cutaneous and neurologic disease associated with HTLV-I infection.
Human T-lymphotropic virus type I (HTLV-I) is the etiologic agent of HTLV-I associated myelopathy (HAM)/tropical spastic paresis (TSP), and adult T-cell leukemia/lymphoma (ATLL). ATLL has been associated with HTLV-I in the southeastern United States. However, to our knowledge, no case reports of HAM/TSP in association with ATLL occurring in the United States have been described. We describe a 40-year-old black woman with a 10-year history of recalcitrant psoriasiform eruption and erythrodermic flares. Medical history is additionally significant for a 2-year history of HTLV-I-associated myelopathy and lower extremity spastic paresis. Polymerase chain reaction with Southern blot analysis was used to detect HTLV-I proviral genome from frozen skin biopsy specimens and peripheral blood mononuclear cells.
Adult↗
Interleukin-8 stimulates leukocyte migration across a monolayer of cultured rabbit gastric epithelial cells. Effect associated with the impairment of gastric epithelial barrier function.
Acute Helicobacter pylori infection produces predominantly neutrophilic infiltration of the gastric mucosa. However, the precise mechanisms and mediators of neutrophil migration are not known. Interleukin-8 (IL-8), a potent chemotactic factor for neutrophils, is present at high concentration in the gastric mucosa of subjects with chronic gastritis caused by H. pylori infection. The aims of this study were to determine whether IL-8 stimulates polymorphonuclear leukocyte (PMN) migration across a cultured monolayer of rabbit gastric epithelial cells and whether PMN migration affects epithelial cell barrier function. Confluent gastric epithelial monolayers grown on the inserts were overlaid with PMNs and various amounts of IL-8 were administered into the well under the insert. Gastric epithelial barrier function was assessed by sodium back diffusion. IL-8 stimulated PMN migration across the monolayer in a dose- and time-dependent manner. PMN transmigration significantly increased sodium back diffusion. In conclusion, IL-8 induces PMN migration across a monolayer of cultured gastric epithelial cells. This IL-8 action is associated with impairment of gastric epithelial barrier function. Since H. pylori infection causes a local mucosal increase of IL-8, our present findings may explain the mechanism of H. pylori-induced PMN infiltration of the gastric glands and mucosal injury.
Animals↗
Recombinant human hepatocyte growth factor facilitates biliary transport after hepatocyte transplantation in Eisai hyperbilirubinemic rats.
Hepatocyte transplantation may offer an attractive treatment for inborn errors of liver metabolism. However, factor(s) are required as stimuli to induce proliferation of the limited number of hepatocytes transplanted. The Eisai hyperbilirubinemic rat (EHBR) is a Sprague-Dawley (SD) mutant rat with conjugated hyperbilirubinemia. EHBRs have impaired canalicular excretory transport of organic anions, bile acid glucuronide, and sulfate. Recombinant human hepatocyte growth factor (rhHGF) (100 microg/kg) was injected intravenously at 2-hr intervals for 10 hr, immediately and 35 days following the intraportal injection of 1 x 10(7) wild-type SD rat hepatocytes. Serum bilirubin concentrations decreased significantly within 35 days and were maintained at significantly reduced levels for 120 days following transplantation. Biliary excretion was demonstrated by the biliary transport of indocyanine green and sulfobromophthalein sodium into the bile. These results indicate that hepatic transport of bile acid conjugates in EHBRs can be restored by hepatocyte transplantation combined with repeated administration of exogenous rhHGF, in conjunction with functioning of the recipient's excretory biliary system.
Animals↗
Influence of estrogen metabolism on proliferation of human breast cancer.
In order to investigate the influence of estrogen metabolism on human breast cancer, estradiol 2- and 16 alpha-hydroxylase (2- and 16 alpha-OHase) activities were determined in the microsomal fractions of cancer tissues by using reverse phase HPLC. 2-OHase activity was detected in most cancer tissues and noncancerous tissues, but the activity was significantly lower in cancer tissues than in the paired noncancerous tissues (0.01 < p < 0.02). Interestingly the patients without lymph node metastasis had significantly higher 2-OHase activity in cancer tissues than those with lymph node metastasis (0.02 < p < 0.05). No correlation was observed between ER status and 2-OHase activity in cancer tissues. On the other hand, 16 alpha-OHase activity was detected only in one third of the breast cancer tissues examined. The activity was not significantly different from that in noncancerous tissues, although it was relatively higher in ER-positive cancer tissues when compared with that in ER-negative ones (0.05 < p < 0.1). Estrone sulfatase activity measured simultaneously in the cytosol fractions of some specimens was much higher in cancer tissues than in noncancerous tissues (0.02 < p < 0.05). We found, however, no correlation between estrone sulfatase activity and estradiol hydroxylase activity. Taken together, our results suggest that the increase in 2-OHase activity prevents the proliferation of breast cancer and that estradiol metabolism is regulated independently of the local biosynthesis of estrogen.
Aryl Hydrocarbon Hydroxylases↗
Effects of insulin on rat renal microvessels: studies in the isolated perfused hydronephrotic kidney.
Although insulin is demonstrated to decrease vascular tone, the role of insulin in renal microcirculation has not been fully determined. In the present study, the effect of insulin on renal microvascular tone was assessed using the isolated perfused hydronephrotic rat kidney. Insulin (300 microU/ml) had no effect on the basal renal microvessel diameter. In addition, insulin did not alter myogenic (that is, pressure-induced) constriction of preglomerular microvessels, with similar magnitude of constriction of preglomerular microvessels, with similar magnitude of constriction observed in response to elevated renal perfusion pressure from 80 to 180 mm Hg (interlobular artery, -23 +/- 3% vs. -19 +/- 4%; afferent arteriole, -22 +/- 3% vs. -21 +/- 4%, for control and insulin, respectively). In striking contrast, insulin dose-dependently reversed the norepinephrine (NE)-induced tone of interlobular arteries, afferent arterioles, and efferent arterioles, with 94 +/- 9%, 104 +/- 6%, and 86 +/- 10% reversal at 300 microU/ml, respectively. These vasodilator actions were markedly inhibited by N-Arg; in the presence of N-Arg, insulin (300 microU/ml) exerted only a modest dilator action on interlobular arteries (24 +/- 9% reversal), afferent arterioles (23 +/- 10% reversal), and efferent arterioles (14 +/- 9% reversal). A similar renal microvascular responsiveness to insulin was also observed during angiotensin II (Ang II)-induced constriction. In conclusion, the ability of insulin to dilate the renal microvasculature differs, with marked inhibitory action during NE/Ang II-induced constriction and almost no inhibition during myogenic constriction. Furthermore, the present study suggests that the insulin-induced renal vasodilation is mediated by nitric oxide.
Angiotensin II↗
Frequent detection of hepatitis A viral RNA in serum during the early convalescent phase of acute hepatitis A.
The diagnosis of type A hepatitis is performed mainly by immunoglobulin M (IgM) anti-hepatitis A antibody assay, but it has not been established whether there is a correlation between changes in viremia and the clinical course of type A hepatitis. We examined hepatitis A virus (HAV) RNA in the sera from type A and non-A, non-B, non-C acute hepatitis and analyzed the relation of HAV viremia with alanine aminotransferase (ALT) and IgM-HA levels. Two hundred sera from 38 patients with type A acute hepatitis and 20 patients with non-A, non-B, non-C acute hepatitis were examined for the presence of HAV RNA. HAV RNA was detected by nested reverse transcription-polymerase chain reaction (RT-PCR) with primers located at the 5' non-translated region of HAV. HAV RNA was detected in 35 of 38 (92%) type A hepatitis patients and in 60 of 156 (38%) serum samples. In contrast, it was detected in none of 44 serum specimens from 20 non-A, non-B, non-C acute hepatitis patients. In type A hepatitis, the mean ALT level in HAV RNA positive specimens was 1,481 +/- 2,042 (range, 20-10,370) IU/L and that in HAV RNA negative specimens 186 +/- 330 (range, 8-1,698). The positivity of HAV RNA was correlated with the level of transaminase at the time of sample collection. The mean duration from the onset of symptoms to disappearance of HAV RNA was 18 +/- 14 days. The mean titer of IgM-HA in HAV RNA positive cases was 5.0 +/- 1.4, in negative cases 5.7 +/- 1.1, with no statistical difference. Our results indicate that HAV RNA in serum is detectable in the majority of type A hepatitis cases in their early convalescent phase by nested RT-PCR.
Acute Disease↗
Subcellular distribution and targeting of the intracellular chloride channel p64.
p64 is an intracellular chloride channel originally identified in bovine kidney microsomes. Using a combination of immunofluorescent and electron microscopic technique, we demonstrate that p64 resides in the limiting membranes of perinuclear dense core vesicles which appear to be regulated secretory vesicles. Heterologous expression of p64 in PancI cells, a cell type which does not normally express p64, results in targeting to a similar compartment. Mutagenesis experiments demonstrate that both the N- and C-terminal domains of the protein independently contribute to subcellular distribution of the protein. The C-terminal domain functions to prevent expression of p64 on the plasma membrane and the N-terminal domain is necessary to deliver p64 to the appropriate membrane compartment.
Animals↗
Measurement of lactate levels in serum and bile using proton nuclear magnetic resonance in patients with hepatobiliary diseases: its utility in detection of malignancies.
Proton nuclear magnetic resonance (1H-NMR) has been utilized for qualitative and quantitative measurement of the components of nonhomogeneous biological specimens, as it can analyze sensitively the chemical structure of organic compounds without pretreatment of the materials. Levels of lactate in serum and bile were measured by 1H-NMR in healthy volunteers and patients with non-malignant or malignant diseases of the liver and biliary tract, and the usefulness of such measurements for the diagnosis of hepatobiliary malignancies was determined. The mean (+/- SD) serum lactate levels were 0.52 +/- 0.33 mmol/l in five healthy volunteers, 1.38 +/- 1.59 mmol/l in 30 patients with non-malignant diseases and 2.95 +/- 2.00 mmol/l in 21 patients with malignant diseases, the differences among the three groups being significant. Biochemical enzymatic measurement of serum lactate levels revealed no such difference. In bile, the spectrum of lactate was observed in all of 16 patients with malignant diseases, but in none of two healthy volunteers and 12 patients with non-malignant diseases. The mean time required for the measurement was 36.77 min for serum and 6.40 min for bile. The measurement of lactate levels in serum and bile using 1H-NMR may be useful for the detection of hepatobiliary malignancies.
Adult↗
Antitumor effects of 5'-deoxy-5-fluorouridine in combination with recombinant human interleukin 2 on murine colon carcinoma 26.
The antitumor activity of recombinant human interleukin 2 (rIL-2) in combination with 5'-deoxy-5-fluorouridine (doxifluridine; 5'-DFUR) against marine colon carcinoma 26 (Colon 26) was studied. BALB/c mice were treated daily for 15 days with 5'-DFUR, rIL-2 or both, beginning on day 7 after subcutaneous transplantation of Colon 26. While mice treated with 5'-DFUR or rIL-2 alone died of tumor growth with pulmonary metastases within 9 weeks posttransplantation, the survival time was significantly prolonged in mice treated with both 5'-DFUR and rIL-2. Most of the combination-treated animals showed the regression of local tumors and the inhibition of pulmonary metastasis. Histopathologically, many tumor cells were degenerated and necrotized, with marked infiltration of mononuclear cells including large granular lymphocytes (LGLs) with periodic acid-Schiff-positive cytoplasmic granules. The cells were positive for CD3 epsilon, asialo GM1 and NK1.1. Spleen cells from the combination-treated mice showed high activities of natural killer (NK) cytotoxicity as well as growth inhibition of Colon 26 and Meth A fibrosarcoma in mice. The results suggest that the combination therapy of 5'-DFUR plus rIL-2 enhanced non-specific cytotoxicity of LGL/NK cells for Colon 26 in tumor-bearing mice and was effective in the inhibition of tumor growth.
Animals↗