Correlation with larval body size of mRNA levels of growth hormone, growth hormone receptor I and insulin-like growth factor I in larval torafugu Takifugu rubripes.

The full-length of insulin-like growth factor (IGF) complementary (c)DNAs encoded by igf-I and igf-II from torafugu pufferfish Takifugu rubripes were cloned in the present study. The deduced amino acid sequences of the two genes showed c. 80% identity each with those of Igf-I and Igf-II from other teleosts, respectively. Two growth hormone (GH) receptors, ghr1 and ghr2, were also cloned in silico using the T. rubripes Fugu genome database. The transcripts of T. rubripes igf-I were detected in slow muscle, heart, skin, gill, liver and intestine but not in fast muscle, spleen and testis of adult fish, whereas those of igf-II were found in all tissues examined. Subsequently, the accumulated messenger (m)RNA levels of igf-I and igf-II were investigated in an F(2) population derived from a male of an apparent fast-growing T. rubripes strain and a wild female T. rubripes together with those of other growth-related genes encoding Gh, Ghr1 and Ghr2, and with those of prolactin (Prl) and leptin (Lep) previously reported. The accumulated mRNA levels of igf-I, gh and ghr1 were significantly correlated to growth rate at larval stages in the population, but not for those of igf-II, prl, ghr2 and lep. Although it is unclear whether or not this phenotype is directly related to the heredity of the fast-growing strain, the findings suggest that the expression of igf-I, gh and ghr1 is involved in the regulation of growth rate at larval stages in T. rubripes.

Efficacy of periodontal disease and tooth loss to screen for low bone mineral density in Japanese women.

The relationship between oral indicators and bone mineral density (BMD) has been studied by many investigators, with mixed and complex results. The purpose of the present cross-sectional study was to evaluate the associations of periodontal conditions and tooth loss with metacarpal BMD (m-BMD) in a community-based cohort and the usefulness of tooth count as a potential screening tool to detect low BMD. Subjects were 356 Japanese women (171 premenopausal, mean age 37.9+/-8.0 years; 185 postmenopausal, mean age 63.3+/-7.7 years). Periodontal status was evaluated by the Community Periodontal Index of Treatment Needs (CPITN). m-BMD was measured by computerized X-ray densitometry. The proportion of subjects with periodontitis (CPITN 3 or 4) increased as m-BMD decreased. The odds ratio (OR) of osteopenia or osteoporosis in relation to periodontitis was 3.2 (95% confidence interval [CI], 2.0--5.3). After adjustment for age and menopausal status, the OR was 2.0 (95% CI, 1.1--3.7). Among postmenopausal women, those having fewer than 20 teeth were 1.6 times more likely to have low m-BMD than those having more than 20 teeth (chi-square for trend in postmenopausal group, 4.27; P<0.05). Receiver-operating curve (ROC) analysis indicated that number of teeth remaining or CPITN score had a greater than 50/50 chance to correctly identify women with osteoporosis or osteopenia, but the areas under the curve (0.72 and 0.67, respectively) are considered less than highly accurate screening tools. These results indicate that periodontitis and tooth loss after menopause may be useful indicators of m-BMD loss in Japanese women.

Low metacarpal bone density, tooth loss, and periodontal disease in Japanese women.

The relationship between periodontitis and systemic bone mineral density in Japanese women is undetermined. We tested the hypothesis that periodontitis was more frequent in women with low metacarpal bone mineral density (m-BMD). Subjects were 190 Japanese women (89 premenopausal, 101 post-menopausal). Periodontal status was evaluated according to the Community Periodontal Index of Treatment Need (CPITN). M-BMD was measured by computed x-ray densitometry. The proportion of subjects with periodontitis (CPITN > or = 3) increased as m-BMD decreased in pre-menopausal (18.2%, 36.9%, and 66.6% in the normal, borderline, and very low m-BMD groups, p < 0.02) and post-menopausal women (41.5%, 54.8%, 60%, and 68.4% in the normal, borderline, low, and very low m-BMD groups, p < 0.05). Among post-menopausal women, those with very low m-BMD had fewer teeth present than women with normal m-BMD (19.9+/-7.2 vs. 25.1+/-4.1, p < 0.01). These results indicate that m-BMD loss is associated with periodontitis in Japanese women, and with tooth loss after menopause.

Time-resolved high-performance liquid chromatography fluorescence detector using a nanosecond pulsed light source for detecting lanthanide-chelated compounds.

We have constructed a time-resolved fluorescence detection (TRFD) system for the analysis of amino compounds with high-performance liquid chromatography (HPLC) using lanthanide ion chelates. In order to carry out time-resolved measurements, we have employed a nanosecond pulsed xenon-arc lamp as an excitation light source. Amino compounds derivatized by isothiocyanobenzyl-EDTA (IEDTA) with the lanthanide chelate are mixed with an enhancer solution in a post-column manner and detected by TRFD. Taking advantage of a property of the long fluorescence lifetime of the lanthanide chelates, high selectivity against background fluorescence was achieved. In order to demonstrate the usefulness of TRFD, fundamental performance tests were carried out. Details of the system are also described.

Detection of a genetic variant, lysine-->glutamic acid at position 372 of human serum albumin, by capillary electrophoresis and structural identification.

A genetic variant of human serum albumin (alloalbumin) is detected by capillary electrophoresis (CE). Two albumin peaks, which were in the ratio of approximately one, were clearly separated. One of the peaks had the same migration time as normal albumin (Alb A) and the other (Alb X) had a longer migration time. SDS-polyacrylamide gel electrophoresis of CNBr fragments (CB) of Alb X indicated that the amino acid substitution was localized in the CB5 fragment (residue 330-446) of the molecule, because of anomalous migration of CB5 in the gel. The CE mapping of the tryptic peptides from the variant CB5 revealed clearly the existence of a new peptide, and the lack of two normal peptides. The sequence analysis of the variant peptide collected by CE micropreparation showed that the N-terminus of the variant peptide corresponded to that of T49 in Alb A. The substitution site, lysine-->glutamic acid at the position 372, was revealed by sequence determination of the variant peptide purified by reversed-phase HPLC.

Hepatectomy for metastatic renal cell carcinoma.

The experience with hepatectomy for metastatic renal cell carcinoma (RCC) has been very rarely reported, because multiple organ metastases ordinarily coexist when hepatic metastases are found out. Three patients who underwent hepatectomy for metastatic RCC are presented here. Radical nephrectomy was performed for the primary renal lesions in all the patients, and their hepatic metastases were resected simultaneously in one of them with a solitary tumor, and about one month later in two of them with multiple (3 and 6) tumors. These operations produced no distinct complications. The patients with 1 and 3 hepatic metastatic lesions survived without tumor recurrence for 12 and 21 months, respectively, while the patient with 6 hepatic metastatic lesions had tumor-free interval of only 2 months and died 10 months after hepatectomy due to lung metastasis. Hepatectomy may be the only promising treatment for hepatic metastases from RCC, but the indication for surgery should be evaluated according to the number of hepatic metastases.

Relationship between nitric oxide and non-adrenergic non-cholinergic inhibitory nerves in human lower esophageal sphincter.

Nitric oxide (NO) has recently been shown to be a neurotransmitter in the non-adrenergic non-cholinergic (NANC) inhibitory nerves in the gastrointestinal tract. To clarify the the role of NO in the human lower esophageal sphincter (LES), enteric nerve responses in lower esophageal tissue specimens obtained from patients with esophageal cancer (n = 7) and patients with gastric cancer (n = 6) were investigated. A mechanographic technique was used to evaluate in vitro LES muscle responses to electrical field stimulation (EFS) of adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers, including NG-nitro-L-arginine (L-NNA) and L-arginine. Findings were: (1) Cholinergic nerves were those mainly involved in the regulation of enteric- nerve responses to EFS in the steady state, and NANC inhibitory nerves acted on the LES; (2) L-NNA concentration-dependently inhibited the relaxation in response to EFS in the LES; and (3) this inhibitory effect in the LES was reversed by L-arginine. These findings suggest that cholinergic and NANC inhibitory nerves play important roles in regulating contraction and relaxation of the human LES, and that NO plays an important role as a neurotransmitter in NANC inhibitory nerves of the human LES.

Preliminary immunohistochemical new findings in the myenteric plexus of patients with intestinal neuronal dysplasia type B.

In order to investigate the causes of abnormal peristalsis of the colon in intestinal neuronal dysplasia (IND), we studied the structure of the myenteric plexus of IND colon using silver-impregnation (Suzuki's method) as well as the innervation of both IND colons and normal colons using immunofluorescence technique with monoclonal antibodies to synaptic vesicles, and antisera to vasoactive intestinal polypeptide (VIP), substance P (SP), methionine-enkephalin (Met-Enk), and gastrin-releasing peptide (GRP). The following results were obtained. 1) In the IND colon, the number of identifiable myenteric ganglia was decreased. In a few cases of IND, irreversible neuron degeneration can be involved in the pathogenesis of IND. 2) In the IND colon, the distribution and fluorescence intensity of synaptic vesicles coincided with those of peptidergic nerve fibers. In the normal colon, synaptic vesicles were much more numerous in the circular muscle layers than in the longitudinal muscle layers, and the fluorescence intensity of those in the circular muscle layers was stronger than that of those in the longitudinal muscle layers. On the other hand, in IND colon, there were fewer synaptic vesicles in the circular muscle layers, and their fluorescence intensity was weak, while there were many synaptic vesicles in the longitudinal muscle layers, and their fluorescence intensity was strong. 3) Morphological abnormalities may exist in synaptic vesicles in the circular muscle layers of the IND colon. 4) Regarding the peptidergic nerve fibers, in the IND colon, innervation of circular muscle layers by Met-Enk-, GRP- and SP-immunoreactive fibers was reduced, and longitudinal muscles were more strongly innervated by immunoreactive fibers than those in the normal colon. 5) Disturbed innervation of non-adrenergic non-cholinergic excitatory nerves may cause the disturbance of muscle contractions in the IND colon. In addition, an imbalance of peptidergic and synaptic vesicle's innervations in both muscle layers may be related to the abnormal peristalsis of IND colon. Also, morphological abnormalities of synaptic vesicles may be concerned with its abnormal peristalsis.

Optical resolution of phenylthiohydantoin-amino acids by capillary electrophoresis and identification of the phenylthiohydantoin-D-amino acid residue of [D-Ala2]-methionine enkephalin.

This is an initial report to propose a protein sequence analysis system with DL differentiation using capillary electrophoresis (CE). This system consists of a protein sequencer and a CE system. After fractionation of phenyl-thiohydantoin (PTH)-amino acids using a protein sequencer, optical resolution for each PTH-amino acid is performed by CE using some chiral selectors such as digitonin, beta-escin and others. As a model peptide, [D-Ala2]-methionine enkephalin (L-Tyr-D-Ala-Gly-L-Phe-L-Met), was used and the sequence with DL differentiation was determined, with the exception of the fourth amino acid, L-Phe, using our proposed system.

Activation but not inactivation of the MTS1 gene is associated with primary colorectal carcinomas.

Several investigators have reported that alterations of the multiple tumor suppressor 1 (MTS1) gene frequently occurred in certain types of tumors. We examined the expression levels and somatic mutations of the MTS1 gene in primary colorectal carcinomas. The MTS1 transcript was quantitatively detected in 14 of 17 tumors, but in only one case of adjacent normal mucosa. Direct sequencing of the amplified MTS1 gene showed no somatic mutation in 17 primary colorectal carcinomas. These results indicate that inactivation of the MTS1 gene is not involved in pathogenesis of primary colorectal carcinomas, and that its enhanced expression is, instead, associated with primary colorectal carcinomas, especially markedly in the early stage of tumor progression.

[Successful combination chemotherapy for a post-operative gastric cancer patient with multiple liver metastases and elevated CEA].

A 75-year-old male was admitted one year after surgery for advanced gastric cancer. He was diagnosed as having multiple liver metastases with elevated serum CEA level. Combination chemotherapy consisting of THP-ADM, MMC and 5'-DFUR was done in the outpatient clinic. As a result, both multiple metastatic liver tumors and serum CEA level showed a remarkable response.

Electrophysiologic assessments in pudendal and sacral motor nerves after ileal J-pouch-anal anastomosis for patients with ulcerative colitis and adenomatosis coli.

PURPOSE: To clarify neurologic function with respect to external anal sphincter and puborectalis muscles after J configuration ileal J-pouch-anal anastomosis for patients with ulcerative colitis and adenomatosis coli, we examined the terminal motor latency in the pudendal and sacral motor nerve (S2-4). METHODS: Latency of the response in the external anal sphincter muscle following digitally directed transrectal pudendal nerve stimulation (PNTML) and in the puborectalis muscle following transcutaneous magnetic stimulation of the cauda equina at the levels S2-4 (SMNLTSS) were measured in 12 patients with ileal J-pouch-anal anastomosis; they were divided into a group with continence (7 cases) and a group with soiling (5 cases). Results were compared with data obtained from 12 patients before operation and 15 controls. RESULTS: Conduction delay of PNTML and SMNLTSS in patients with soiling was longest, followed by delay in those without any soiling, then delay in patients before operation, and then controls. In addition, significant differences were also noted between conduction delay of PNTML in controls and those who are incontinent and experience soiling (P < 0.05 and P < 0.01, respectively), and there were significant differences also noted between conduction delay of PNTML in patients before operation and those who are incontinent and experiencing soiling (P < 0.05 and P < 0.01, respectively). Conduction delay of PNTML and SMNLTSS were found in patients before operation rather than in controls. No significant differences were noted between conduction delay of PNTML and SMNLTSS in patients before operation and controls. Significant differences were also noted between conduction delay of PNTML and SMNLTSS in patients who are incontinent and experiencing soiling (P < 0.01, respectively). CONCLUSION: These findings support the hypothesis that soiling after this procedure may be partially caused by damage to pudendal and sacral motor nerves (S2-4).

Heterotopic splenic autotransplantation in a neonate with splenic rupture, leading to normal splenic function.

Loss of the spleen in infants and young children frequently results in overwhelming infection. To preserve splenic function, heterotopic splenic autotransplantation after splenectomy was performed on a newborn in whom the spleen was traumatized beyond repair because of birth trauma. Postoperatively, the percentage of pitted red blood cells that accurately reflects splenic reticuloendothelial function was normal. Splenic scintigrams obtained 3 weeks after the operation showed uptake by the reimplanted tissue. The otherwise mandatory long-term antibiotic prophylaxis could be reduced to 3 weeks in the newborn. At 7 years of age, the patient has not shown any increased susceptibility to infections.

[Development of quality of life questionnaire for patients with colorectal cancer in surgical area--a study of reliability and validity of Tokyo Yamabuki Forum Version].

We developed a new questionnaire in the surgical area based on a core quality of life (QOL) questionnaire for patients with gastrointestinal cancer. In this study, we investigated the validity and reliability of a QOL questionnaire (Tokyo Yamabuki Forum Version) for patients with colorectal cancer. The questionnaire was composed of 17 items including 5 scales (basic sensory scale, psychological scale, physiological scale, defection-related scale and active scale) and a face scale as an global scale. The time needed to answer questionnaires was expected to be around 7 minutes and the questionnaires should basically be answered by the patients themselves everyday in the hospital. The study was performed in 10 hospitals in the Tokyo area, and 394 samples collected from 21 patients with rectal and colonic cancers were analyzed. A number of respondents failed to answer the question "Do you feel your foods tasty?", so we judged this item inappropriate and deleted it from the analysis. Fifteen items, including 5 scales showed satisfactory internal consistency and construct validity in correlation and factor analyses. Performance status showed a low correlation between each item, each scale and the global scale, while SDS and STAI showed an inordinately negative correlation with the fundamental and physical scales. Especially, SDS revealed an extremely close correlation with the active scale, and STAI showed an excessive correlation with the psychological scale. In the time course of QOL under chemotherapy, reductions (aggravations) were observed in both the total score of 15 items and global scale within one week postoperatively, but after that recovered to preoperative levels at 2 weeks postoperatively. A tendency to QOL improvement was observed 2 weeks after starting chemotherapy or chemoimmunotherapy. QOL of 13 patients was measured over 3 months, and the longest term was 8 months. The results suggested that this QOL questionnaire has sufficient reliability and validity to be usable for patients with colorectal cancer in the surgical area and that this model is applicable for long-term QOL surveys and frequent measurement.

Epidural metastasis in chemoresistant Wilms' tumor with perilobar nephroblastomatosis.

An 8-year-old boy with vertebral and epidural metastases was diagnosed with Wilms' tumor associated with perilobar nephroblastomatosis (NB) based on histologic examination. During combined chemotherapy with vincristine, actinomycin D, doxorubicin, and cyclophosphamide (NWTS-3 J protocol), a rapid increase in tumor size was observed. The treatment was replaced with etoposide and carboplatin (JET regimen). A transient response was sustained for 5 months during this chemotherapy. However, regrowth of the tumor was observed and the patient died 11 months after the initial chemotherapy.

Unique presentation of heterotopic pancreatic tissue arising from small-bowel mesentery.

Heterotopic pancreatic tissue in the small-bowel mesentery is an extremely rare anomaly: no such case has been reported in the English literature. We report a 5-year-old boy in whom the aberrant tissue formed a retroperitoneal cyst. The condition was further complicated by a ventral pancreatic duct defect. The unique clinical manifestations and difficulty in making the diagnosis are discussed.

Biological characteristics of neuroblastoma with spontaneous tumor reduction: a case report.

The authors examined the biological characteristics of a neuroblastoma with spontaneous tumor reduction. A 6-month-old boy with a pelvic neuroblastoma underwent surgical extirpation of the tumor 1 month after diagnosis. The size of the tumor reduced spontaneously while he was awaiting operation. The low proliferative activity of the tumor cells and the presence of apoptosis in the tumor tissue were shown by an immunohistochemical method using anti-PCNA antibody and a DNA fragmentation analysis, respectively. These results suggest that the spontaneous tumor reduction seen in this patient may well be caused by the overwhelming apoptosis of tumor cells.

A role of nitric oxide in Hirschsprung's disease.

Nitric oxide (NO) has recently been shown to be a neurotransmitter in the nonadrenergic noncholinergic (NANC) inhibitory nerves in the gastrointestinal tract. To clarify the significance of NO in Hirschsprung's disease (HD), enteric nerve responses in colonic tissue obtained from HD patients were investigated. Colonic tissue specimens were obtained from four patients with HD and from 11 patients without constipation who were used as controls. A mechanograph was used to evaluate in vitro colonic responses to electrical field stimulation (EFS) of the adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers, and NG-nitro-L-arginine (L-NNA) and L-arginine with the following results: (1) NANC inhibitory nerves were found to act on normal human colon, but had no effect on aganglionic colon; (2) L-NNA concentration dependently inhibited the relaxation in response to EFS in the normal colon, but had no effect on aganglionic colon; and (3) this inhibitory effect was reversed by L-arginine in the normal colon, but had no effect on the aganglionic colon. Nitric oxide mediates the relaxation reaction of NANC inhibitory nerves in the human colon, but the effect of NO was absent in aganglionic colon. The loss of action by NO may be implicated in the impaired motility observed in aganglionic colon.

Terminal motor latency in the pudendal nerves after colectomy with mucosal proctectomy and ileal J pouch-anal anastomosis for ulcerative colitis.

Pudendal nerve terminal motor latencies (PNTML) were measured in eight patients with ulcerative colitis who underwent colectomy with mucosal proctectomy and ileal J pouch-anal anastomosis, using a new digitally directed transrectal stimulation and recording technique, and the results were compared with data obtained from 15 control subjects. The conduction delay of PNTML in the patients with some degree of fecal incontinence was the longest, followed by those without any incontinence, and then the control subjects. These findings support the hypothesis that fecal incontinence after this procedure may be partially caused by damage to the pudendal nerve.