Vascular endothelial growth factor in dialysate in relation to intensity of peritoneal inflammation.

BACKGROUND: Peritoneal inflammation may induce changes in peritoneal microvessels, including neoangiogenesis/vasculogenesis, leading to increased peritoneal solute transport rate (PSTR) and loss of ultrafiltration capacity. We hypothesized that an inflammatory reaction in the peritoneal cavity during peritonitis induces increased synthesis of vascular endothelial growth factor (VEGF). We therefore studied the relationship between peritoneal inflammation markers, VEGF, and transport of fluid and solutes in rats during acute peritoneal inflammation induced by lipopolysaccharide (LPS) added to standard glucose-based dialysis solution. METHODS: Under ether anesthesia, male Wistar rats were injected intraperitoneally with 30 mL Dianeal 3.86% without (Control; n=6) or with LPS (microg/mL): 0.001 (LPS 0.001; n=6), 0.01 (LPS 0.01; n=7), 0.1 (LPS 0.1; n=7), 1.0 (LPS 1.0; n=8). After 8 hours, dialysate volume (IPV), peritoneal solute transport rate (PSTR) and dialysate cell count (DCC) were measured and effluent samples were collected. RESULTS: LPS i.p. resulted in increased PSTR and decreased IPV (p<0.005). DCC (cells/microL) and the neutrophil/macrophage ratio were higher for all LPS concentrations compared to the control group. After 8 hours, LPS-exposed rats had significantly higher dialysate levels of all investigated cytokines (TNF-alfa, MCP-1 and IL-10) than the control group. Addition of LPS resulted in increased dialysate VEGF concentrations (pg/mL) (LPS 0.001, 28.2+/-5.9; LPS 0.01, 38.9+/-11.6; LPS 0.1, 43.0+/-5.9; LPS 1.0, 46.6+/-11.3; Control, 14.5+/-9.8; p<0.0005 for all LPS vs. Control). CONCLUSIONS: The infusion of Dianeal 3.86% with different doses of LPS induced a strong acute intraperitoneal inflammatory reaction with increased DCC and cytokine levels, resulting in increased peritoneal solute transport and decreased IPV. LPS induced a dose-dependent parallel increase of the intraperitoneal concentrations of MCP-1, IL-10 and TNF-alfa, as well as of VEGF. These results suggest that intraperitoneal VEGF synthesis is induced in response to inflammation, and that this may be an important component in the process leading to peritoneal transport alterations.

The relationship between lifestyle, social characteristics and obesity in 3-year-old Japanese children.

It has been observed that obese children receive genetic and environmental effects that are associated with them being overweight. With regard to the latter, lifestyles such as eating habits and physical activity have been focused on. In the present study, the social characteristics which would dominate their lifestyles were investigated as background variables. For this purpose, 9668 Japanese children aged three years who were all born in Toyama prefecture, Japan, in 1998, served as birth cohort subjects. For the comparison between obese (Kaup Index; mass in kg/(height in m)2 > or = 18) and nonobese (Kaup index < 18) children, irregular snack intake, physical inactivity and reduced sleeping hours were chosen as statistically significant obesity-related lifestyle indicators for the children. For social characteristics, family construction (expanded family with grandparents/nonexpanded family), main caregiver (mother/other), attending a nursery school (yes/no) and mother's employment (full-time worker/other) were chosen. These were significantly associated with the obesity-related lifestyles mentioned above using multiple logistic regression analysis adjusted for other variables of social characteristics as well as for gender and birth month (July-December/January-June). The two greatest population-attributable risk percentages were observed for mother as main caregiver (-36.5%) and attending a nursery school (-28.9%) for irregular snack intake. Therefore, these two social characteristics substantially reduced the number of children with irregular snack intake. On the other hand, the two social characteristics were reversed in children with reduced sleeping hours (population-attributable risk percentage of mother as main caregiver: 15.4%; attending a nursery school: 17%). In contrast with favourable effects on snack intake these social characteristics showed an adverse influence on the sleeping habits of children.

Molecular mechanisms of promoter regulation of the gp34 gene that is trans-activated by an oncoprotein Tax of human T cell leukemia virus type I.

We investigated the molecular mechanism of transcriptional activation of the gp34 gene by the Tax oncoprotein of human T cell leukemia virus type I (HTLV-I). gp34 is a type II transmembrane molecule belonging to the tumor necrosis factor family and is constitutively expressed on HTLV-I-producing cells but not normal resting T cells. The transcriptional regulatory region of the gp34 gene was activated by HTLV-I Tax in the human T cell line Jurkat, in which endogenous gp34 is induced by Tax. Sequence analysis demonstrated that two NF-kappaB-like elements (1 and 2) were present in the regulatory region. Both NF-kappaB-like elements were able to bind to NF-kappaB or its related factor(s) in a Tax-dependent manner. Chloramphenicol acetyltransferase assays indicated that NF-kappaB-like element 1 was Tax-responsive, although the activity was lower than that the native promoter. NF-kappaB-like element 2 elevated promoter activity when combined with NF-kappaB-like element 1, indicating cooperative function of the elements for maximum promoter function. Unlike typical NF-kappaB elements, the NF-kappaB-like elements in gp34 were not activated by treatment of Jurkat cells with phorbol ester despite induction of the NF-kappaB-like binding activity. Chloramphenicol acetyltransferase reporter assays using the region upstream of the NF-kappaB-like elements identified an upstream region that reduced transcription from cognate and noncognate core promoters in a Tax-independent manner. Our results imply complex regulation of expression of the gp34 gene and suggest implication of gp34 in proliferation of HTLV-I infected T cells.

Serological and histological features of anti-HCV-positive individuals without serum HCV RNA.

Antibody to hepatitis C virus (anti-HCV) in patients who are negative for HCV RNA in serum may indicate a memory of past infection of HCV. However, their clinical features have not been well understood. Fourteen anti-HCV-positive but HCV RNA-negative individuals were examined for serological and histological features. As a result, it was found that they had only antibody to HCV core antigen C22-3 with or without antibody to nonstructural viral antigen C33c on a recombinant immunoblot assay (RIBA), and that an concentration of anti-C22 was low. Liver biopsy showed two having no evidence of an obvious hepatic injury, two having a minimal change, and two having portal fibrosis. HCV RNA was not found in the liver. These results corroborate an idea that the anti-HCV in HCV RNA-negative individuals implies a past infection of HCV. Furthermore, it is suggested that a combination of an appearance pattern of antibody to HCV antigens on RIBA and anti-C22 titer are an useful marker to distinguish anti-HCV-positive individuals without viremia from those with viremia.

Helicobacter pylori may cause "reflux" gastritis after gastrectomy.

Patients with "reflux" gastritis after gastrectomy suffer from a variety of symptoms, and this type of gastritis may sometimes compromise the quality of life of these patients. Since Helicobacter pylori is considered to be one of the most important pathogenetic factors in gastritis, the association between H. pylori and reflux gastritis was investigated in this study. A total of 145 patients with gastrectomy were entered into the study. Five biopsy specimens from the gastric remnant were taken at upper gastrointestinal endoscopy. One specimen was examined pathohistologically, and the remaining four were examined for H. pylori infection. Fifty-two patients (36%) demonstrated H. pylori infection. The prevalence of H. pylori was significantly higher in patients who had a partial gastrectomy, and it was significantly lower in patients who had undergone gastrectomy more than 4 years previously. The histologic gastritis score in patients with H. pylori infection was significantly higher. Furthermore, H. pylori was eradicated in patients with some symptoms of gastritis and no bile reflux to the residual stomach at endoscopy; in these patients the symptoms were relieved and the histologic gastritis score decreased significantly. In conclusion, possible involvement of H. pylori is suspected in the pathogenesis of "nonreflux" gastritis after gastrectomy.

Etiological involvement of Helicobacter pylori in "reflux" gastritis after gastrectomy.

OBJECTIVES: "Reflux" gastritis after gastrectomy is associated with various symptoms that are often detrimental to the patients' quality of life. However, prevention of the reflux does not always bring relief from the symptoms of gastritis. Helicobacter pylori (H. pylori) is now considered one of the most important pathogenetic factors in gastritis. The association between H. pylori infection and reflux gastritis after gastrectomy was investigated in the present study. METHODS: In total, 115 patients who had undergone gastrectomy were entered in this study. Five biopsy specimens from the gastric remnant were taken during upper GI endoscopy. One specimen was examined pathohistologically, and the remaining four were examined for H. pylori infection. The histological degree of gastritis was determined according to the score system of Rauws et al. RESULTS: Forty-six patients (40%) demonstrated H. pylori infection in their stomachs. The prevalence of the infection was significantly higher in patients with conventional gastrectomy than in those with subtotal gastrectomy. The prevalence of H. pylori infection was significantly lower in patients who had undergone gastrectomy more than 4 yr ago. The histological gastritis score in patients with H. pylori infection was significantly higher than in those without H. pylori infection. Furthermore, the eradication of H. pylori in patients with both serious gastritis symptoms and no bile reflux improved the symptoms and significantly decreased the histological gastritis score. CONCLUSIONS: The results suggest that H. pylori is a factor in the pathogenesis of reflux gastritis after gastrectomy.

[Residual gastritis after gastrectomy and Helicobacter pylori--its clinical significance].

Residual gastritis after gastrectomy brings the various symptoms such as abdominal pain, nausea, emesis and loss of appetite, and often hazards quality of life of the patient. Bile reflux to the stomach is believed as one of the important pathogenesis of residual gastritis, however the prevention for bile reflux cannot always heal the gastritis. Helicobacter pylori (H. pylori) is considered as one of the most important pathogenesis of gastroduodenal ulcer and gastritis, and H. pylori may possibly cause residual gastritis after gastrectomy. However, the association between infection with H. pylori and the residual gastritis has not revealed yet. In the present study, the association with H. pylori and the residual gastritis after gastrectomy was investigated in 56 patients who had undergone gastrectomy before. Twenty-four patients (42.9%) had H. pylori infection at their stomachs and the incidence of the infection in the patients with gastrectomy was significantly higher with subtotal gastrectomy. As for the histological gastritis score of Rauws (Rauws' score), Rauws' score of H. pylori positive group was significantly higher than H. pylori negative group. Furthermore, the eradication of H. pylori for the patients with serious symptoms of gastritis improved the symptoms and decreased significantly Rauws' score. These results suggest that H. pylori was associated with the pathogenesis of residual gastritis after gastrectomy.

Synergism between two distinct elements of the HTLV-I enhancer during activation by the trans-activator of HTLV-I.

We have conducted functional studies of the enhancer elements of human T-cell leukemia virus type I (HTLV-I) using the human T-cell lines Jurkat and MOLT 4, which are negative for HTLV-I, and MT-2 and TL-Mor, which carry the proviral genome of HTLV-I. Two distinct elements have been implicated in function of the HTLV-I enhancer. One is the 21-base-pair (bp) core element that is responsible for trans-activation by the HTLV-I trans-activator p40tax and that has the ability to bind to cyclic-AMP responsive element binding factor (CREB)-like factor(s). The other is a region interposed between the 21-bp elements. In this study we demonstrate that a subfragment (C26) in the region between the 21-bp elements is involved in trans-activation by p40tax, possibly through binding to an NF-kappa B-like nuclear factor or factors. Formation of the protein-DNA complex with the C26 subfragment was positively affected by p40tax. The C26 element conferred partial responsiveness to p40tax when linked to one copy of the 21-bp element that, by itself, showed little activation in response to p40tax. However, the C26 element alone, even when repeated, could not be activated by p40tax, unlike other NF-kappa B-binding elements. In contrast, the C26 element itself was profoundly activated upon stimulation with 12-O-tetradecanoylphorbol-13-acetate. These findings therefore suggest that the HTLV-I enhancer contains multiple functional elements, including binding sites for at least CREB- and NF-kappa B-like factors, which synergistically cooperate in activation of the HTLV-I enhancer in response to p40tax. Our results also demonstrate that TPA-dependent activation of the HTLV-I enhancer may be mediated through the C26 element.

Molecular cloning and characterization of a novel glycoprotein, gp34, that is specifically induced by the human T-cell leukemia virus type I transactivator p40tax.

We have cloned and sequenced a cDNA encoding gp34, a novel glycoprotein expressed in cells bearing human T-cell leukemia virus type I (HTLV-I). HTLV-I has a trans-acting transcriptional activator, p40tax, that is thought to be implicated in leukemogenesis through the activation of cellular enhancers. With a subline (JPX-9) of the human T-cell line Jurkat, in which p40tax is inducible, gp34 was shown to be of cellular origin and to be transcriptionally activated by p40tax. It was also demonstrated that two species of mRNA are generated from one copy of the gp34 gene and that these mRNAs encode the identical gp34 product and differ in the 3' untranslated region. Analysis of the deduced amino acid sequence of gp34 showed that it lacks typical signal peptides; however, it has a hydrophobic stretch for membrane anchoring and four possible N-linked glycosylation sites at the carboxy-terminal portion, indicating that it belongs to the family of membrane proteins whose carboxy-terminal portion protrudes out of the cell. The gp34 gene displayed relatively delayed induction compared with other genes activated by p40tax. Taken together with the observation of the dependence of gp34 expression on HTLV-I p40tax, unlike other p40tax-dependent genes such as those for the interleukin-2 receptor alpha chain and c-fos, which are expressed or induced under physiological conditions, we predict that the mechanism involved in the induction of gp34 expression by p40tax is distinct from and more intricate than those for the previously characterized genes.

[Combination chemotherapy with etoposide, ADM, and CDDP (EAP) for advanced gastric cancer].

EAP therapy has been performed on 50 cases of advanced gastric cancer from January 1988 to September 1989. Adriamycin 20 mg/m2, Cisplatin 40 mg/m2 and Etoposide 100 mg/m2 were administered on day 1 and 7, 2 and 8, and 4, 5 and 6, respectively, with not less than 2 courses every 3 to 4 weeks. Complete success, PR, NC and PD were obtained in 48, 21, 20 and 7 cases, respectively, the rate of effectiveness being 43.8% with a confidence interval 95% of 30-58%. The rate of effectiveness by lesions for evaluation was high (30.4, 100, and 50% for primary lesion, Virchow's lymphnodal metastasis and liver metastasis, respectively). MST was 5.1 months for EAP therapy, which was highly effective but led to no prolonged survival period. Thus, it is thought that good control of leukopenia, a dose limiting factor remains to be examined.

[Loco-regional cancer chemotherapy with a new drug delivery system, "anticancer drug-fibrin clot"].

Five different types of anticancer drugs were individually entrapped into fibrin clots using our own material, "G.T.XIII" to provide an "anticancer drug-fibrin clot" for regional cancer chemotherapy. Anticancer drugs used in the present study were ADM, MMC, MTX, 5-FU and cDDP. The release of drugs from fibrin clots was studied in vitro. Each fibrin clot was intraperitoneally administered to cancer (AH-130)-bearing rats to evaluate the oncolytic effects. The activities of anticancer drugs delivered from the clots were maintained for more than two weeks. Survival terms of cancer bearing rats were remarkably prolonged with the anticancer drug-fibrin clots. Neither recurrence of ascites nor metastases of malignant cells was observed in the rats treated with such clots. Our newly devised anticancer drug-fibrin clots showed a sustained release of oncolytic drugs and favorable antineoplastic effects. This newly devised drug delivery system suggested a clinical potential for regional cancer chemotherapy.

Protein phosphorylation mediated by IL-2/IL-2 receptor beta-chain interaction.

We previously found that IL-2 rapidly induced protein phosphorylation of a 67-kDa (pp67) and four 63-kDa (pp63s) cellular proteins in various T cells. Here, we show that the IL-2-stimulated phosphorylation is mediated by the IL-2R beta-chain composed of the high affinity IL-2R, and induced by activation of Ca2+/phospholipid-dependent protein kinase C (PKC). The IL-2-stimulated phosphorylation was always observed in various T cell lines bearing high affinity IL-2R, but never observed in cells which express only low affinity IL-2R consisted of alpha-chain alone. When the expression of high affinity IL-2R was modified by anti-IL-2R mAb for reducing the affinity to 8- to 10-fold lower without affecting the sites of IL-2R, the effective dose of IL-2 on phosphorylation of pp67 increased 8 to 10 times. When cells were treated with pronase, approximately 95% sites of low affinity IL-2R were selectively decreased, but the IL-2 dose dependency for pp67 phosphorylation was little affected. These data exactly suggest that protein phosphorylation in response to IL-2 such as pp67 and pp63s, is mediated by high affinity but not low affinity IL-2R. Furthermore, the IL-2-stimulated phosphorylation of these proteins was also observed in MLA 144 cells which express only low affinity IL-2R consisting of beta-chain alone. In addition, various phorbol esters and tumor promoters, which activate PKC, were also demonstrated to induce the phosphorylation of a pp67 and pp63s in these T cell lines. Therefore, the present study suggests that IL-2/IL-2R beta-chain interaction triggers the phosphorylation of pp67 and pp63s, where the PKC may have an important role.

[Intra-arterial infusion chemotherapy with adriamycin for the treatment of locally advanced breast cancer--studies of serum and tissue concentrations of adriamycin].

The effect of intra-arterial infusion chemotherapy with adriamycin (ADM), with special references to its pharmacokinetic behavior and also the tissue concentrations, was studied in 8 patients with locally advanced breast cancer. A high clinical response rate (CR + PR) of 70.0% was obtained by the treatment, and also remarkable degenerative changes of tumor cells were histologically noted in 5 out of 9 surgical specimens (55.6%). Side effects such as alopecia (100%), leukopenia (87.5%) and stomatitis (85.5%) were frequently observed, but none of them delayed the following mastectomy. Pharmacokinetic studies of intra-arterial administration of ADM showed a low serum concentration and its rapid clearance from serum. ADM also seemed to have a great affinity to the regional lymph nodes (1.21 micrograms/g tissue), compared to cancer (0.61 micrograms/g) and normal breast tissues (0.51 micrograms/g). These results suggested that intra-arterial infusion chemotherapy with ADM might be applicable not only for the preoperative treatment of locally advanced breast cancer, but also for the control of lymph node metastasis.

[Changes in immunological capacity of patients with cancer of the large intestine].

The changes of host immunity during the cancer treatment were studied in 27 patients with large bowel cancer. As the result, compared with preoperative status, increasing tendency of serum IgG, IgA levels, normalization of T cell per cent and remarkably lowered CH50 level were found postoperatively. However, in the patients treated with immunochemotherapy, no definite changes were found during the treatment. It was suggested that the CH50 level might have close relation to the curativity or the prognosis in these operative cases.