Oral administration of a recombinant cholera toxin B subunit promotes mucosal healing in the colon.

Cholera toxin B subunit (CTB) is a component of a licensed oral cholera vaccine. However, CTB has pleiotropic immunomodulatory effects whose impacts on the gut are not fully understood. Here, we found that oral administration in mice of a plant-made recombinant CTB (CTBp) significantly increased several immune cell populations in the colon lamina propria. Global gene expression analysis revealed that CTBp had more pronounced impacts on the colon than the small intestine, with significant activation of TGFß-mediated pathways in the colon epithelium. The clinical relevance of CTBp-induced impacts on colonic mucosa was examined. In a human colon epithelial model using Caco2 cells, CTBp, but not the non-GM1-binding mutant G33D-CTBp, induced TGFß-mediated wound healing. In a dextran sodium sulfate (DSS) acute colitis mouse model, oral administration of CTBp protected against colon mucosal damage as manifested by mitigated body weight loss, decreased histopathological scores, and blunted escalation of inflammatory cytokine levels while inducing wound healing-related genes. Furthermore, biweekly oral administration of CTBp significantly reduced disease severity and tumorigenesis in the azoxymethane/DSS model of ulcerative colitis and colon cancer. Altogether, these results demonstrate CTBp's ability to enhance mucosal healing in the colon, highlighting its potential application in ulcerative colitis therapy besides cholera vaccination.

Exome sequencing for bipolar disorder points to roles of de novo loss-of-function and protein-altering mutations.

Although numerous genetic studies have been conducted for bipolar disorder (BD), its genetic architecture remains elusive. Here we perform, to the best of our knowledge, the first trio-based exome sequencing study for BD to investigate potential roles of de novo mutations in the disease etiology. We identified 71 de novo point mutations and one de novo copy-number mutation in 79 BD probands. Among the genes hit by de novo loss-of-function (LOF; nonsense, splice site or frameshift) or protein-altering (LOF, missense and inframe indel) mutations, we found significant enrichment of genes highly intolerant (first percentile of intolerant genes assessed by Residual Variation Intolerance Score) to protein-altering variants in general population, an observation that is also reported in autism and schizophrenia. When we performed a joint analysis using the data of schizoaffective disorder in published studies, we found global enrichment of de novo LOF and protein-altering mutations in the combined group of bipolar I and schizoaffective disorders. Considering relationship between de novo mutations and clinical phenotypes, we observed significantly earlier disease onset among the BD probands with de novo protein-altering mutations when compared with non-carriers. Gene ontology enrichment analysis of genes hit by de novo protein-altering mutations in bipolar I and schizoaffective disorders did not identify any significant enrichment. These results of exploratory analyses collectively point to the roles of de novo LOF and protein-altering mutations in the etiology of bipolar disorder and warrant further large-scale studies.

Cord blood immune biomarkers in small for gestational age births.

Fetal growth restriction is a risk factor for development of adulthood diseases, but the biological mechanism of this association remains unknown. Limited biomarkers have been studied in settings of preterm birth and maternal inflammation, but the relationship between a wide range of immune biomarkers and fetal growth has not been studied. The hypothesis of this study was that fetal growth restriction is associated with altered immune biomarker levels. We examined the relationship between small for gestational age (SGA) status and 27 umbilical cord blood immune biomarkers. This study was part of a large-scale cohort study of preterm birth and low birth weight conducted at Boston Medical Center, an inner city, predominantly minority patient population. Growth status was determined based on birth weight standardized to an internal reference. There were 74 SGA births and 319 appropriate for age (AGA) births with complete clinical and biomarker data. Adjusting for covariates and using AGA as reference, SGA births had lower levels of log IL-1ß (ng/l; ß -0.38, 95% CI -0.57, -0.19, P < 0.01), log BDNF (ß -0.29, 95% CI -0.55, -0.03, P < 0.05) and log NT-3 (ß -0.46, 95% CI -0.77, -0.15, P < 0.01). No associations were found between other biomarkers and SGA. In conclusion, three biomarkers were selectively associated with SGA status. Our results provide information that could be used to guide additional studied aimed at determining mechanisms that contribute to fetal growth.

Design and production of genetically modified soybean protein with anti-hypertensive activity by incorporating potent analogue of ovokinin(2-7).

The potent anti-hypertensive peptide, RPLKPW, has been designed based on the structure of ovokinin(2-7). The sequence encoding this peptide was introduced into three homologous sites in the gene for soybean beta-conglycinin alpha' subunit. The native alpha' subunit as well as the modified, RPLKPW-containing alpha' subunit were expressed in Escherichia coli, recovered from the soluble fraction and then purified by ion-exchange chromatography. The RPLKPW peptide was released from recombinant RPLKPW-containing alpha' subunit after in vitro digestion by trypsin and chymotrypsin. Moreover, the undigested RPLKPW-containing alpha' subunit given orally at a dose of 10 mg/kg exerted an anti-hypertensive effect in spontaneously hypertensive rats, unlike the native alpha' subunit. These results provide evidence for the first time that a physiologically active peptide introduced into a food protein by site-directed mutagenesis could practically function in vivo even at a low dose.

Designing potent derivatives of ovokinin(2-7), an anti-hypertensive peptide derived from ovalbumin.

We obtained a potent anti-hypertensive peptide, RPFHPF, by replacing the amino acid residues of ovokinin(2-7) (RADHPF), an orally active anti-hypertensive peptide derived from ovalbumin. After intravenous administration in anesthetized Wistar rats, the designed peptide [Pro2, Phe3]-ovokinin(2-7) had a long-lasting hypotensive activity at a dose of 10 mg/kg, while that of ovokinin(2-7) was only transient even at a dose of 100 mg/kg. After oral administration in conscious spontaneously hypertensive rats (SHRs), [Pro2, Phe3]-ovokinin(2-7) significantly lowered the systolic blood pressure in a dose-dependent manner. It is noteworthy that the minimum effective dose of [Pro2, Phe3]-ovokinin(2-7) was 0.3 mg/kg, about one-thirtieth of that of ovokinin(2-7). On the other hand, orally administered [Pro2, Phe3]-ovokinin(2-7) did not show any significant hypotensive effect in normotensive Wistar-Kyoto rats (WKYs) even at a dose of 3 mg/kg. Taken together, [Pro2, Phe3]-ovokinin(2-7) proved to be an ideal, potent anti-hypertensive peptide with little effect on normal blood pressure when given orally.

Bioactive peptides derived from food proteins preventing lifestyle-related diseases.

Many kinds of bioactive peptides which might prevent lifestyle-related diseases are released from food proteins after enzymatic digestion. Inhibitory peptides for angiotensin I-converting enzyme (ACE) having anti-hypertensive effect have been isolated from enzymatic digests of various food proteins. LKPNM, which was isolated from the thermolysin digest of dried bonito was activated 8-fold by ACE itself and showed a prolonged effect after oral administration. Two vasorelaxing peptides, ovokinin and ovokinin(2-7), showing antihypertensive effect after oral administration were obtained from ovalbumin digests. We found that low molecular weight peptides derived from food proteins lowered serum cholesterol without increasing excretion of cholesterol and bile acids. An immunostimulating peptide isolated from an enzymatic digest of soybean protein prevented alopecia induced by cancer chemotherapy.

A novel anti-hypertensive peptide derived from ovalbumin induces nitric oxide-mediated vasorelaxation in an isolated SHR mesenteric artery.

In this report, we deal with the isolation of a novel vasorelaxing peptide from a chymotryptic digest of ovalbumin and its vasorelaxing activities. This peptide is composed of Arg-Ala-Asp-His-Pro-Phe (RADHPF) in its sequence, corresponding to residues 359-364 of ovalbumin. This peptide (30-300 microM) exerted a dose-dependent vasodilation in an isolated mesenteric artery from a spontaneously hypertensive rat which was pre-constricted by phenylephrine, besides the relaxation being endothelium-dependent. It is noteworthy that the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester inhibited this relaxation, implying involvement of nitric oxide in its mechanism of action. Following oral administration of RADHPF at a dose of 10 mg/kg, the systolic blood pressure in a spontaneously hypertensive rat was significantly lowered.

Gastroduodenal intussusception secondary to a gastric carcinoma.

A case of gastroduodenal intussusception secondary to transpyloric prolapse of a primary gastric carcinoma is reported. Both the condition itself and the leading tumor, gastric carcinoma, are extremely exceptional. A diagnosis of this rare entity was established pre-operatively by endoscopy that demonstrated spontaneous reduction of the intussusception. This may be the first documentation of spontaneous reduction of the gastroduodenal intussusception during endoscopy.

Malignant mesenchymoma arising in an incisional scar of the abdominal wall.

A case of malignant mesenchymoma of the anterior abdominal wall, composed of leiomyosarcoma, chondrosarcoma and osteosarcoma is described. The tumour, arising in the old scar of a previous operation, was completely removed surgically. Eleven months after the operation the patient is well with no evidence of recurrence. There have been no previous reports of malignant mesenchymoma developing in operation scars. We believe that this traumatic factor played a significant role in the aetiology of this rare soft-tissue neoplasm.

[A case of advanced gastric cancer with liver metastasis disappeared by oral administration of 5'-deoxy-5-fluorouridine after absolute non-curative operation].

A 64-year-old male was found to have a gastric cancer (Borrmann type 3) on the posterior wall of the stomach. Histological diagnosis was poorly differentiated adenocarcinoma. Ultrasonography revealed a hypoechoic lesion (18 x 14 mm) in the anteroinferior segment (S5) of the liver. The biopsy specimens from those lesions proved to be poorly differentiated adenocarcinoma. Total gastrectomy and distal pancreatosplenectomy were performed. During the operation, other liver metastases were found in the anterosuperior segment (S8) and inferolateral segment (S3). 5'-deoxy-5-fluorouridine (800 mg/day) was administered per os from the 33 rd postoperative day. The hypoechoic lesion of S5 in liver disappeared in the 17th postoperative month. The man is healthy with no evidence of recurrence 47 months after the operation. This case is very important because of the long-term CR.

[Peritoneal dissemination of cecal cancer disappearing in a patient taking 5'-DFUR for 3 years].

A 51-year-old woman presenting with a left ovarian tumor underwent left oophorectomy and hysterectomy with intraperitoneal administration of cisplatin, cyclophosphamide, and 5-fluorouracil on December 13, 1989. Microscopically the tumor was well-differentiated adenocarcinoma. Postoperatively cecal cancer was detected. Laparotomy revealed multiple peritoneal dissemination on March 23, 1990, when palliative ileocecal resection was performed. Microscopy revealed several of the nodules to be adenocarcinoma and the cecal lesion well differentiated adenocarcinoma. The patient was then placed on 5'-deoxy-5-fluorouridine (800 subsequently 600 mg/day). In November, 1991, bright red blood per rectum led to the discovery of rectal cancer. On January 26, 1993, laparotomy revealed rectal tumor invading the bladder but the metastatic nodules on the peritoneum had disappeared. Thus pelvic exenteration was performed. The tumor was well-differentiated adenocarcinoma. The ovarian lesion may have been a metastasis from the cecal cancer, a so-called Krukenberg tumor. The rectal cancer was most likely to be asynchronous cancer. 5'-DFUR seemed effective in remitting the peritoneal dissemination.

SPECT images of technetium-99m-ethyl cysteinate dimer in cerebrovascular diseases: comparison with other cerebral perfusion tracers and PET.

UNLABELLED: To assess the clinical role of 99mTc-ethyl cysteinate dimer (ECD) as a cerebral perfusion tracer, 10 patients with unilateral cerebral infarction were studied. METHODS: ECD SPECT images were compared with IMP and/or HMPAO SPECT in nine patients, seven in chronic phases and two in subacute phases. Five of these patients and one additional patient with chronic infarction received PET imaging in order to compare ECD distribution with quantitative regional cerebral blood flow (rCBF) and oxygen metabolism (rCMRO2) images. RESULTS: In patients with chronic cerebral infarction, regression analysis showed excellent correlation between ECD and IMP in the uptake ratio of lesions-to-nonaffected cortices (r = 0.91). In two cases of subacute infarction, decreased uptake of ECD was observed in the area of "luxury perfusion," which showed elevated or preserved rCBF with diminished rCMRO2. On visual analysis, ECD image contrast was less prominent compared to that of IMP, but contrast was better than that of HMPAO. CONCLUSIONS: ECD uptake showed a curvilinear relationship against rCBF, suggesting flow-limited uptake in a high flow range. ECD is a clinically useful cerebral perfusion tracer with distinct characteristics when compared with other available agents.

PET and the autoradiographic method with continuous inhalation of oxygen-15-gas: theoretical analysis and comparison with conventional steady-state methods.

The steady-state method using 15O gas inhalation and positron emission tomography (PET) is a simple and practical way of imaging cerebral blood flow (CBF) and oxygen metabolism. Several disadvantages do exist, however, including prolonged examination time, requirement of steady-state and a large tissue heterogeneity effect. To avoid the drawbacks of the steady-state method but to preserve its simplicity, we applied the PET/autoradiographic method to the build-up phase during the continuous inhalation of 15O-gas with intermittent arterial sampling. A simulation study was performed to determine the optimal scanning period, evaluate the delay and dispersion effect of the input function and estimate the tissue heterogeneity effect. To assess the clinical feasibility of the proposed technique for the study of oxygen metabolism, sequential measurements with this method and the conventional steady-state method were performed in eight patients. The simulation study showed that a 5-min scan started 3 min after the commencement of 15O-gas inhalation was optimal. With this method, the delay and dispersion effect on CBF was the same as that of the conventional steady-state method, but the tissue heterogeneity effect was reduced. In eight patients, CBF values calculated by this method showed time dependency and were slightly higher than those obtained by the steady-state method. The oxygen extraction fraction showed no significant time dependency and was well correlated with that obtained by the steady-state method. We conclude that the proposed method is a simple and acceptable alternative to the conventional steady-state method.

Improved detection rate of early breast cancer in mass screening combined with mammography.

A cohort study to compare mass screening with and without mammography was conducted in Miyagi Prefecture, Japan in order to establish whether the effectiveness of breast cancer screening would be improved when mammography was combined with physical examination. A trial of mass screening combined with mammography was carried out in 9634 women aged over 50. Lateral imaging of the breast using single-view film mammography was performed at the first stage of mass screening in addition to physical examination of the breast. Results in the trial were compared with those obtained in 35511 age-matched subjects without mammography. Thirty breast cancers were found in the trial with a detection rate of 0.31%, which was much higher than that (0.08%) obtained by physical examination without mammography. In 15 of the 30 patients the breast tumor was not palpated at the first screening, but abnormal findings were detected in the mammography. A higher rate (73%) of early breast cancer was obtained in the screening trial with mammography than that (39%) obtained in the screening with physical examination alone. Mass screening combined with mammography is superior to that without mammography for breast cancer screening, especially for the detection of non-palpable, early breast cancer.

[99mTc-HMPAO SPECT in the brain death--a case report].

Brain single photon emission computed tomography (SPECT) with 99mTc-d,l-hexamethyl-propyleneamine oxime (HMPAO) was performed twice in a 78-year-old man clinically diagnosed as brain death according to the standard criteria of the Japanese Ministry of Welfare. The first brain SPECT demonstrated the tracer accumulation in the brain, indicating preserved cerebral blood flow. The second brain SPECT performed 3 days later revealed cessation of the blood flow. In patients with preserved cerebral blood flow, the diagnosis of brain death cannot be made, even if they meet the existing criteria, because previous report noted the recovery in some of those patients. Brain perfusion SPECT plays an important role as a confirmatory test for the diagnosis of the brain death.

Noninvasive measurement of regional cerebral blood flow change with H(2)(15)O and positron emission tomography using a mechanical injector and a standard arterial input function.

To estimate changes in regional cerebral blood flow (rCBF) without arterial sampling in the study of functional-anatomical correlations in the human brain, using (15)O-labeled water and PET, a standard arterial input function was generated from the input function in 10 normal volunteers with dose calibration and peak time normalization. The speed and volume of injection were precisely controlled with a mechanical injector. After global normalization of each tissue activity image, the standard arterial input function was applied to obtain estimated CBF images. Relative changes in estimated rCBF to whole brain mean CBF(DeltaFest) and those in regional tissue activity (DeltaC) were compared with true relative rCBF changes (DeltaF) in 40 pairs of images obtained from 6 normal volunteers. DeltaFest correlated well with DeltaF, whereas DeltaC consistently underestimated DeltaF. This noninvasive method simplifies the activation studies and provides the accurate estimation of relative flow changes.

Lumbar artery pseudoaneurysm: an unusual cause of a retroperitoneal hematoma: report of a case.

Pseudoaneurysm of the lumbar artery is a rare complication of penetrating trauma. We present herein a case thought to have been caused by a blow to the left flank without any evidence of a stab wound. In this patient, the diagnosis of a first lumbar artery pseudoaneurysm with a retroperitoneal hematoma was confirmed by computed tomography (CT) findings, after which transcatheter embolization was successfully performed.

Induction of discriminant function concerning postoperative local recurrence or distant metastasis in 589 patients with differentiated thyroid cancer.

For the purpose of predicting postoperative local recurrence and/or distant metastasis in differentiated thyroid cancer, a discriminant analysis was done. To assess the reliability of this method and the induced function, simulation was done by using random samples under various conditions. Simulation revealed that the results obtained by this method were relatively stable. The discriminant functions Z1 and Z2 were induced, which are required at many hospitals. If the ratio of recurrence is less than 1:8 in a hospital, function Z1 should be applied. If the ratio is 1:10, function Z2 should be applied. Z1 = -(sex x 2.104) + (age x 0.032) - (diameter of tumor x 0.033)-(site of tumor x 0.871) - (histology x 0.393) + 0.249 (discriminant boundary 0.451) Z2 = -(sex x 1.967) + (age x 0.018) + (diameter of tumor x 0.025) - (site of tumor x 0.646) + (histology x 0.014) - (local invasion x 0.840) - (lymph node metastasis x 0.009) + (operation x 0.364) - (lymph node dissection x 0.235) + 1.058 (discriminant boundary 0.400). In an internal check, sensitivity was 78%, specificity 64%; and false-negative rate 5%. In an external check, sensitivity was 68%, specificity 93%, and false-negative rate 9%. In the actual application to hospitals, sensitivity was 58% to 100%, specificity 47% to 63%, and false-negative rate 0% to 16%. In clinical usage, a high sensitivity and low false-negative rate are required. These results fulfilled this condition.

Analysis of thyroid carcinoma based on material registered in Japan during 1977-1986 with special reference to predominance of papillary type.

BACKGROUND: As geographic differences have been observed in the characteristics of thyroid carcinoma, an analysis was made on thyroid carcinoma in the iodine rich country of Japan. METHODS: A total of 10,973 patients with histologically confirmed thyroid carcinoma registered in Japan from 1977-1986 were analyzed. Cases detected incidentally at autopsy and cases of nonepithelial tumor were excluded. This series included approximately 27% of all thyroid carcinoma cases in Japan. RESULTS AND CONCLUSIONS: Histologic distribution showed that papillary carcinoma accounted for 78.4% of cases, follicular carcinoma accounted for 17.2%, medullary carcinoma for 1.4%, squamous cell carcinoma for 0.3%, and anaplastic carcinoma for 2.7%. There is a tendency in Japan to diagnose papillary carcinoma as follicular carcinoma and to diagnose malignant lymphoma as anaplastic carcinoma. It was considered that the percentage of papillary carcinoma was higher and the percentage of follicular carcinoma and anaplastic carcinoma was lower than foregoing values. The characteristics of thyroid carcinoma in Japan were described, and the low incidence of nonpapillary carcinoma compared with papillary carcinoma was discussed in relation to iodine excess as an etiologic factor.