T-bet over-expression regulates aryl hydrocarbon receptor-mediated T helper type 17 differentiation through an interferon (IFN)γ-independent pathway.

Various transcription factors are also known to enhance or suppress T helper type 17 (Th17) differentiation. We have shown previously that the development of collagen-induced arthritis was suppressed in T-bet transgenic (T-bet Tg) mice, and T-bet seemed to suppress Th17 differentiation through an interferon (IFN)-γ-independent pathway, although the precise mechanism remains to be clarified. The present study was designed to investigate further the mechanisms involved in the regulation of Th17 differentiation by T-bet over-expression, and we found the new relationship between T-bet and aryl hydrocarbon receptor (AHR). Both T-bet Tg mice and IFN-γ-/- -over-expressing T-bet (T-bet Tg/IFN-γ-/- ) mice showed inhibition of retinoic acid-related orphan receptor (ROR)γt expression and IL-17 production by CD4+ T cells cultured under conditions that promote Th-17 differentiation, and decreased IL-6 receptor (IL-6R) expression and signal transducer and activator of transcription-3 (STAT-3) phosphorylation in CD4+ T cells. The mRNA expression of ahr and rorc were suppressed in CD4+ T cells cultured under Th-17 conditions from T-bet Tg mice and T-bet Tg/IFN-γ-/- mice. CD4+ T cells of wild-type (WT) and IFN-γ-/- mice transduced with T-bet-expressing retrovirus also showed inhibition of IL-17 production, whereas T-bet transduction had no effect on IL-6R expression and STAT-3 phosphorylation. Interestingly, the mRNA expression of ahr and rorc were suppressed in CD4+ T cells with T-bet transduction cultured under Th17 conditions. The enhancement of interleukin (IL)-17 production from CD4+ T cells by the addition of AHR ligand with Th17 conditions was cancelled by T-bet over-expression. Our findings suggest that T-bet over-expression-induced suppression of Th17 differentiation is mediated through IFN-γ-independent AHR suppression.

Improvement of rice (Oryza sativa L.) seed oil quality through introduction of a soybean microsomal omega-3 fatty acid desaturase gene.

Microsomal omega-3 fatty acid desaturase is an essential enzyme in the production of the n-3 polyunsaturated fatty acid alpha-linolenic acid during the seed developing stage. We have constructed a chimeric gene consisting of a maize Ubi1-P-int and a soybean GmFAD3 cDNA, which was introduced into rice plants by Agrobacterium-mediated transformation. Ten transformants containing the chimeric gene were established and expression subsequently confirmed by Northern blotting. Furthermore, alpha-linolenic acid content of the T(1) seeds increased dramatically up to tenfold that of the control, and this phenotype was also stably inherited in the T(2) and T(3) progenies. These results demonstrate that the alpha-linolenic acid content of rice seed oil can easily be altered using the combination of a high-activity promoter and a GmFAD3 gene.

Effect of hot water extract from Agaricus blazei Murill on antibody-producing cells in mice.

We investigated the immunopotentiating activities of boiled water-soluble extracts from desiccated Agaricus blazei Murill (ABM). Effect of ABM extract on antibody production was investigated by method of hemolytic plaque-forming cells (PFC) against sheep red blood cells (SRBC) antigen. ABM extracts significantly (p<0.01) increased the number of PFC in spleen with intraperitoneal administration at doses of 25 mg/kg as compared with control group. The populations of Mac-1- or CD25-positive cells significantly (p<0.01, p<0.001) increased, but in CD19-positive cells, there were no differences in ABM-treated mice as compared with control mice. The expressions of IL-6 and IL-1beta mRNA were augmented by ABM extract in both peritoneal macrophages and spleen cells. These results suggested that ABM extract might be an effective stimulator for T cell and macrophage to IL-1beta and IL-6 release, resulting in augmentation of antibody production against SRBC antigen.

Analysis of cooking oil fumes by ultraviolet spectrometry and gas chromatography-mass spectrometry.

This paper investigated the components, especially aldehydes, in the fume condensates from four kinds of cooking oil using ultraviolet (UV) spectrometry and gas chromatography-mass spectrometry (GC-MS). It was observed that there was a great change of the UV absorption spectra from the results of the unheated oil to the results of the fume after heat treatment (190-200, 230-240, and 270-280 degrees C). There was a strong peak within the wavelength range of 260-270 nm in each condensate sample. From the GC-MS results, it was tentatively deduced that there were some 2,4-dialkylenaldehydes and other conjugated compounds in the condensates. The results showed there were large amounts of hexanal and 2-heptenal in the cooking oil fume and that the total aldehyde peak areas of the condensates from four kinds of oil were around 30-50% of the total peak area at 270-280 degrees C.

[Experience of four cases of postoperative pulmonary embolism requiring surgical treatment].

Four patients underwent a pulmonary embolectomy using cardiopulmonary bypass for acute pulmonary embolism which had occurred after various operations. In two cases, dehydration due to either diabetes insipidus or ileus had existed. In two cases, pulmonary embolism suddenly occurred in our hospital. In the remainder, the disease occurred in the previous hospitals and its diagnosis was established on the 6th and 7th postoperative days, respectively. In massive pulmonary embolism, echocardiography and/or enhanced chest CT are useful for prompt and noninvasive diagnosis. Thrombolytic therapy was performed in only one case before surgical embolectomy, which was not effective. Three patients were discharged without any postoperative complications, but one requiring preoperative external cardiac massage died of multiple organ failure 9 days after operation. Acute pulmonary embolism is one of the fatal postoperative complications. Recognition of this entity, and prompt diagnosis and treatment are essential for managing the fatal disease. Even in the early postoperative period, embolectomy using cardiopulmonary bypass is a safe and effective treatment.

Molecular cloning and characterization of phosphatidylcholine transfer protein-like protein gene expressed in murine haploid germ cells.

We have isolated a cDNA clone specifically expressed in spermiogenesis from a subtracted cDNA library of mouse testis. The cDNA consisted of 1392 nucleotides and had an open reading frame of 873 nucleotides encoding a protein of 291 amino acid residues. Computer-mediated homology search revealed that the nucleotide sequence was unique but the deduced amino acid sequence had similarity to mouse phosphatidylcholine transfer protein (PCTP). We named this newly isolated gene PCTP-like protein. Northern blot analysis revealed a 1.4-kilobase mRNA expressed in the testis, kidney, liver, and intestine with the highest level in the testis. Messenger RNA expression in the testis was detected first on Day 23 in postnatal development and then increased up to adulthood. The protein, having a molecular weight of approximately 40 000, was encoded by the mRNA and was detected at the tail of the elongated spermatids and sperm by immunohistochemical staining.

Control of DAF-7 TGF-(alpha) expression and neuronal process development by a receptor tyrosine kinase KIN-8 in Caenorhabditis elegans.

KIN-8 in C. elegans is highly homologous to human ROR-1 and 2 receptor tyrosine kinases of unknown functions. These kinases belong to a new subfamily related to the Trk subfamily. A kin-8 promoter::gfp fusion gene was expressed in ASI and many other neurons as well as in pharyngeal and head muscles. A kin-8 deletion mutant was isolated and showed constitutive dauer larva formation (Daf-c) phenotype: about half of the F(1) progeny became dauer larvae when they were cultivated on an old lawn of E. coli as food. Among the cells expressing kin-8::gfp, only ASI sensory neurons are known to express DAF-7 TGF-(beta), a key molecule preventing dauer larva formation. In the kin-8 deletion mutant, expression of daf-7::gfp in ASI was greatly reduced, dye-filling in ASI was specifically lost and ASI sensory processes did not completely extend into the amphid pore. The Daf-c phenotype was suppressed by daf-7 cDNA expression or a daf-3 null mutation. ASI-directed expression of kin-8 cDNA under the daf-7 promoter or expression by a heat shock promoter rescued the dye-filling defect, but not the Daf-c phenotype, of the kin-8 mutant. These results show that the kin-8 mutation causes the Daf-c phenotype through reduction of the daf-7 gene expression and that KIN-8 function is cell-autonomous for the dye-filling in ASI. KIN-8 is required for the process development of ASI, and also involved in promotion of daf-7 expression through a physiological or developmental function.

[Direct effects of Chinese herbal medicine "hochuekkito" on sperm movement].

BACKGROUND AND PURPOSE: Chinese herbal medicine, "Hochuekkito" is widely used for male infertility in Japan. There have been many reports concerning its clinical usefulness but very few reports of in vitro experiments studying the mechanism of its effects. In addition to stimulating germ cells, we analyzed its direct effects on sperm using computer assisted semen analyzer (CASA). MATERIALS AND METHODS: Motile sperm were prepared using swim up technique from semen collected from ten healthy volunteers. Sperm movements (motility, velocity, linearity) were analyzed by CASA after adding either serum containing anti-sperm antibody (ASA) or normal serum with or without Hochuekkito. RESULTS: Two hours after adding serum with ASA, the decrease of sperm motility was significantly reduced from 25.1% (92.8%-->67.7%) to 12.5% (92.9%-->80.6%) by adding Hochuekkito. No significant difference in velocity and linearity was observed between two groups. By adding normal serum, any of three parameters differed significantly with or without Hochuekkito. CONCLUSION: Protective effects of Hochuekkito on sperm was suggested. Although normal sperm with ASA was used in this report, since the sperm of infertile patients are said to be more fragile, this results imply that direct protective effect is one of the mechanism of Hochuekkito for male infertility.

A case of drug eruption caused by the crude drug Boi (Sinomenium stem/Sinomeni caulis et Rhizoma).

We report a case of drug eruption caused by the crude drug Boi. A 41-year-old female patient had been diagnosed with chronic rheumatoid arthritis in the department of internal medicine. After ingestion of a decoction of the crude drug Boi for the alleviation of arthralgia, a slight fever developed, which was followed by systemic edematous erythema with itching. HPLC showed that the main components of the crude drug Boi are sinomenine and magnoflorine. The results of patch tests were negative for all oral drugs that the patient had been taking. Oral ingestion tests showed that the patient showed positive reactions to the as-is Boi boiling-water decoction and 1/10-volume sinomenine. Based on this, the drug eruption was judged to be caused by sinomenine. It is considered the first time that the causative component of a drug eruption was confirmed by oral ingestion tests with components of a crude drug of Kampo medicine (Sino-Japanese traditional medicine).

Colonic submucosal tumors: a new classification based on radiologic characteristics.

OBJECTIVE: Despite advances in gastrointestinal radiologic techniques, findings from barium enema studies cannot be used reliably to predict the histologic nature of a colonic submucosal tumor. Therefore, we generated a new and comprehensive classification system for all colonic submucosal tumors based on a correlation between their radiologic characteristics and their histopathologic features. MATERIALS AND METHODS: Between 1978 and 1990, 89 patients had 95 colonic submucosal tumors diagnosed on the basis of barium enema studies and confirmed by biopsy or surgery. Of these, 39 patients (44%) were symptomatic and 50 patients (56%) were asymptomatic. Single lesions (82 cases) and multiple lesions (12 cases of lymphoma and one case of blue rubber bleb nevus) were observed. The 95 lesions included 21 carcinoids, 27 malignant lymphomas, 17 lipomas, 10 lymphangiomas, six leiomyosarcomas, five leiomyomas, and nine others. We analyzed the radiologic findings and classified these lesions according to their appearance. RESULTS: All tumors were classified on the basis of their morphologic appearance into five types: (1) wide-based sessile lesion with gradually sloping margin and smooth surface (17 cases); (2) wide-based sessile lesion, more polypoid than the first type, with a smooth surface, with or without a central depression (49 cases); (3) wide-based sessile lesion with lobulated surface (six cases); (4) pedunculated lesion with smooth or granular surface (17 cases); (5) unclassified, not any of the previously mentioned types, may be diffusely stenotic or aneurysmal (six cases). The most common finding was the wide-based sessile lesion with a smooth surface and a possible central depression; next most common was the wide-based sessile lesion with gradually sloping margin and smooth surface; next was the pedunculated lesion with a smooth or granular surface. Eighty-six percent of the lesions were single; 14% were multiple. Histologic confirmation revealed mainly lymphoma (29%) and carcinoid (22%), of which the most common finding was a wide-based sessile lesion with a smooth surface, with or without a central depression. CONCLUSION: This new classification based on radiologic characteristics covers all varieties of colonic submucosal tumors. It can serve as a guideline for predicting a specific diagnosis of a colonic submucosal tumor on the basis of radiologic findings.

Hypogonadism of male prolactinomas: relation to pulsatile secretion of LH.

In order to investigate whether a hypothalamic disorder cause hypogonadism in male prolactinomas, LH pulsatile secretion was studied in 13 male patients. Serum PRL levels ranged from 186 to 45,000 ng ml-1 before treatment, and all the tumors were macroadenomas. Reduced LH secretion was revealed in 5 of 13 patients, and FSH was reduced in 1 of 13. Serum testosterone (T) levels were lower than the normal limit in all the patients. HCG tests in 3 patients showed good responses, but the peak values of T were lower than those of normal men. LH pulsatilities were examined in 5 hyperprolactinemic patients before treatment, in 4 hyperprolactinemic patients after operation, and in 8 normoprolactinemic patients after operation and/or bromocriptine treatment. There was no significant difference of the mean LH values, the frequencies of LH pulses, and amplitudes among the hyperprolactinemic patients before operation (n = 5), the normoprolactinemic patients after operation (n = 8), and normal men (n = 7). From these results, it was evident that the hypothalamus and pituitary function of male prolactinomas were well preserved, in spite of higher serum PRL levels and larger tumor size than those reported in females. It is suggested that the main cause of hypogonadism in these patients is due to testicular dysfunction resulting from excessive serum PRL.

Effect of retinoic acid on proliferation of estrogen-responsive transformed murine Leydig cells in serum-free culture.

B-1 F cells, one of the sublines established from mouse Leydig cell tumor, have been found to be maintained as an estrogen-responsive cell line under the serum-free culture conditions. Reported results that retinoids have action mechanisms similar to those of estrogen prompted us to examine the effect of retinoids on the proliferation of B-1 F cells. Stimulation of B-1 F cell growth by retinoic acid in a dose-dependent manner was observed, whereas retinoic acid did not promote but inhibited the proliferation of MCF-7 cells (estrogen- and retinoic acid receptor-positive human breast cancer cells). To elucidate the mechanism of retinoic acid-dependent cell growth, simultaneous treatment with retinoic acid and estradiol was carried out. The result did not show the additive effect on B-1 F cell growth. Hydroxytamoxifen, a potent antiestrogen, inhibited not only estradiol-dependent but also retinoic acid-dependent cell growth. However, retinoic acid failed to be associated with estrogen receptor, suggesting that retinoic acid induced enhancement of B-1 cell growth through its interaction with retinoic acid receptor. Northern blot analyses of polyadenylated RNA with complementary DNA probes for human retinoic acid receptor alpha, beta, and gamma revealed the presence of transcripts encoded by retinoic acid receptor alpha gene in B-1 F cells. These results would suggest that enhancement of the B-1 F cell growth is mediated through interaction of retinoic acid with retinoic acid receptor alpha. This stimulatory activity is inhibited by estrogen receptor complexed with hydroxytamoxifen.

[Diagnosis of colon cancer (depth of invasion) by barium enema examination; present condition].

Accuracy of the barium enema examinations in diagnosis of the depth of invasion of the small carcinomas of the colon was reviewed. The materials included 101 sessile carcinomas and 12 small advanced carcinomas that were 20 mm or less in diameter. The carcinomas with stalks and the depressed type early carcinomas (Type IIc) were excluded from the study, because the former were impossible to the nature by X-ray examinations and the latter were extremely rare. Presence of the depressed portions on the surface of the elevated lesions observed in the frontal views and the degree of the deformity observed in the profile views of the lesions were correlated with the pathological specimen and the accuracy of the radiological diagnosis of the depth of invasion was estimated. The cases with gross malignant cell invasion into the submucosal or the deeper layers were diagnosed correctly with present radiological diagnostic criteria. But, those with a small amount of malignant cell invasion into the submucosal layer or the cases with malignant cell invasion limited to the mucosal layer (intramural cancer) were not always correctly diagnosed and there was a variety in diagnosis of the depth of invasion in this group. With the current radiological techniques, proper profile views of the lesions were not obtained in quite a few cases, nor the surface characteristics of the small elevated lesions were well documented. It would probably be possible to improve these techniques and to improve diagnostic accuracy to some extent, but with certain limitations.

Epidermal grafting using the tops of suction blisters in the treatment of vitiligo.

Fourteen patients with nondermatomal and 31 patients with segmental vitiligo were treated by epidermal grafting using the tops of suction blisters. Epidermis of the vitiliginous area was removed by a blister induced by methoxsalen plus ultraviolet A, and the tops of suction blisters were placed next to one another, resembling paving stones. Reepithelialization was completed in about two weeks, and skin color normalized in most cases about half a year later. Repigmentation persisted in most cases of segmental vitiligo, whereas loss of pigmentation occurred again in nonsegmental vitiligo. Scars were not observed on either the graft site or the donor site.

[Intraoperative autotransfusion in aortic aneurysm operations combined with temporary external shunt].

Intraoperative autotransfusion was done in nine patients who underwent major vascular surgery for thoracic aortic aneurysms including one total abdominal aneurysm using temporal external shunt. After general heparinization (1mg/kg), shed blood was collected by Sorenson Autotransfusion System with local heparinization. After confirming ACT longer than 300-400 second, 425-3700 ml, averaged 1804 ml, of shed blood was reinfused to the patients by gravity flow. The reinfusion rate of shed blood was 43.1 +/- 15.8%. There were no deaths and no complications referable to autotransfusion. Although mild hemolysis was observed immediately after surgery, plasma free hemoglobin level returned to normal in the first operative day and no renal failure occurred. The volume of bleeding after surgery was not differed from that of control. Microembolism was not evident clinically. We concluded that intraoperative autotransfusion is safe and useful procedure in major vascular surgery if proper anticoagulation is done.

[Case of lipohyperplasia of the ileocecal valve with adenocarcinoma of the ascending colon].

A rare case of lipohyperplasia of the ileocecal valve contiguous with adenocarcinoma of the ascending colon is reported. The patient was a 67-year-old female with a chief complaint of muco-bloody stool. Barium enema X-ray study revealed a filling defect in the proximal portion of the ascending colon, suggestive of Borrmann II-type carcinoma and enlargement of the ileocecal valve. Ileocecotomy and right colectomy were performed. Histological examination disclosed that the tumor of the ascending colon consisted of well differentiated adenocarcinoma; the proliferation of fat tissue in the submucosa of the ileocecal valve was diagnosed as lipohyperplasia of the ileocecal valve. The histogenesis of this lipohyperplasia seems to be secondary development induced by the adenocarcinoma of the ascending colon.