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This study was performed to clarify the critical role of optimal levels of nitric oxide on fecundity in mice during the implantation period. Mature female pregnant mice were treated with either nitric oxide donor molsidomine (3, 15, 60 mg kg(-1)) or nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester (L-name; 0.3, 1.5, 6 mg kg(-1)) every 12 h, seven times from the night of Day 2 to Day 5 of gestation. They were killed on Day 14 of gestation. Pregnancy rates in each group (n = 22) and the number of live or absorbed fetuses in each mouse was calculated. The pregnancy rates in the experimental group were reduced in a dose-dependent manner. The rate in the control group was 100%, whereas those in the 60-mg molsidomine and 6-mg L-name groups were 40.9 and 31.8%, respectively. Histological analysis of uteri on Day 5 of gestation after treatment with 60 mg molsidomine or 6 mg L-name suggested retarded decidualization of stromal cells or defective function of predecidualized cells. In conclusion, optimal levels of nitric oxide are crucial for endometrial function and embryo implantation.

Endogenous gibberellins (GAs) in the young leaves and xylem sap of tea plants (Camellia sinensis L.) were analyzed by GC-MS. The following GAs were identified by comparing their mass spectra and KRIs with those of authentic specimens: GA9 and GA20 in the leaves; GA9, GA12, GA15, GA20, GA44, GA51 and GA53 in the xylem sap.

Background: Recent studies demonstrated not only that advanced glycation end product could be found in amyloid tissue from patient with dialysis related amyloidosis, but also that amyloid beta2-microglobulin was modified with N(epsilon)-(carboxymethyl)lysine (CML). We wanted to determine if CML could be a biomarker in these patients., Methods: To raise polyclonal anti-carboxymethyllysine antibody, human serum albumin was carboxymethylated by glyoxylic acid and was immunized to rabbits as antigen. Carboxymethyllysine-hemoglobin (CML-Hb) levels were measured by the dot blotting method using this antibody., Results: The levels of CML-Hb were 6.68 +/- 3.10 nmol CML/mg Hb in nondiabetic hemodialysis patients (N = 70), 6.39 +/- 3.43 nmol CML/mg Hb in diabetic hemodialysis patient (N = 21), and 3.13 +/- 0.88 nmol CML/mg Hb in 47 healthy volunteers. For clinical signs of dialysis-related amyloidosis, 70 nondiabetic hemodialysis patients were scored according Gejyo's criteria. The CML-Hb levels in patients with a high amyloid score as well as a low amyloid score were significantly higher than in patients with negative amyloid score (8.89 +/- 3.53 nmol CMLmg Hb, 7.28 +/- 2.32 nmol CML/mg Hb vs. 5.11 +/- 2.09 nmol CML/mg Hb, P < 0.001, P < 0.05). Furthermore, the CML-Hb levels correlated significantly with serum values of the methylguanidine over creatinine ratio and hyaluronate., Conclusions: We suggest that CML-Hb is increased in blood from patients on maintenance hemodialysis and is thus a potential biomarker of oxidative damage in these patients. Moreover, CML-modification of protein may play a pathogenic role in the development of dialysis related amyloidosis.

Fluorodeoxyglucose positron emission tomography (FDG-PET) has been utilized since glucose metabolism in cancer tissue is considered to relate to its malignant potential. We evaluate the application of FDG-PET to prostate cancer. FDG uptake of cancer tissues was higher with higher Gleason grade, advanced clinical stage and higher serum PSA value than that with lower Gleason grade, localized clinical stage and lower serum PSA value. The measurement of glucose metabolism in the prostate using FDG-PET was suggested to be useful for evaluation of the biological malignant potential of prostate cancer. It may serve as a predictive factor for prognosis and may be useful in determining the effectiveness of hormonal therapy in prostate cancer.

Ectopic expression of the homeobox gene, NTH15 (Nicotiana tabacum homeobox 15) in transgenic tobacco leads to abnormal leaf and flower morphology, accompanied by a decrease in the content of the active gibberellin, GA1. Quantitative analysis of intermediates in the GA biosynthetic pathway revealed that the step from GA19 to GA20 was blocked in transgenic tobacco plants overexpressing NTH15. To investigate the relationship between the expression of NTH15 and genes involved in GA biosynthesis, we isolated three cDNA clones from tobacco encoding two types of GA 20-oxidase and a 3 beta-hydroxylase. RNA gel blot analysis revealed that the expression of one gene (Ntc12, encoding GA 20-oxidase), which in wild-type tobacco plants was abundantly expressed in leaves, was strongly suppressed in the transformants. The expression level of Ntc12 decreased with increasing severity of phenotype of transgenic tobacco leaves. The abnormal leaf morphology was largely overcome by treatment with GA20 or GA1 but not by GA19. These data strongly suggest that overexpression of NTH15 inhibits the expression of Ntc12, resulting in reduced levels of active GA and abnormal leaf morphology in transgenic tobacco plants. In situ hybridization in wild-type tobacco revealed that expression of Ntc12 occurred mainly in the rib meristem, cells surrounding the procambium and in leaf primordia. Expression was not seen in the tunica, corpus and procambium, tissues in which NTH15 was predominantly expressed. The contrasting expression patterns of these genes may reflect their antagonistic functions in the formation of lateral organs from the shoot apical meristem.

Legionellosis is an important cause of severe pneumonia in the community. Inadequate therapy will lead to respiratory distress syndrome, disseminated intravascular coagulation (DIC) and finally fatal multiple organ failure. We encountered a rare case in which early manifestation included septic shock and DIC complicated by acute myocardial infarction (AMI) suspected to be derived from Legionnaires' disease. A 54-year-old healthy female complained of lumbago, high fever and dry cough 10 days after visiting a hot spring spa. She was emmergently admitted due to shock. Physical examination demonstrated hypotension, high fever, course creakle in the right lower lung. Hepatosplenomegaly, lymphadenopathy and eruption were not found. WBC count was 34600/microliters with nuclear shift. CRP elevated. FDP, D dimer and TAT also elevated CPK elevated with dominance of the MB isozyme. Chest roentogenography revealed congestive heart failure, pleural effusion and obscure pneumonic shadow and EKG showed ST segment elevation in leads I, II, III, aVF, V4, V5, and V6. The patient was diagnosed as having septic shock, DIC and AMI. She was treated with gabexate mesilate, high dose methyl prednisolone and dopamine hydrochloride as well as piperacillin, meropenem, isepamycin and fluconzaole. Despite intensive care, the blood pressure fell again and pneumonia had progressed on the 8th hospital day. These antibiotics appeared to be ineffective. Erythromycin was then administered and a dramatic effect. was obtained as the patient recovered. Serum titer of Legionella pneumophila (serogroup 1) rose to 128-fold 2 weeks after the onset. Other serum titers such as Chlamydia psittaci, Rickettsia, Mycoplasma were all negative. Cultures obtained from the sputum, throat swab, urine and blood did not yield any microorganisms. Although the diagnosis could not be confirmed because the titer did not elevate over 256-fold of 4-fold within 2 weeks after the onset, Legionella infection was highly suspected from the clinical features. This is a rare case in which septic shock and DIC with AMI preceded pulmonary symptoms in a non-immunocompromised patient.

A 76-year-old-man was admitted to our hospital for a recurrent bladder tumor. He had received intravesical bacillus Calmette-Guérin (BCG) treatment for a transitional cell carcinoma of the bladder. A follow-up cystoscopy revealed a solitary papillary tumor in the left bladder wall. A transurethral cold cup biopsy revealed a nephrogenic adenoma without any evidence of malignant cells. We discuss the pathogenesis of nephrogenic adenoma and suggest that prolonged cystitis caused by intravesical BCG may play an etiological role.

Although the precise pathoetiology of Behçet's disease (BD) remains obscure, patients with BD have a high incidence of chronic infectious foci, indicating an enhanced susceptibility to chronic tonsillitis, and dental caries. Sometimes, clinical symptoms appear after treatment of these foci in BD patients. It is believed that BD might be related to an allergic reaction to a bacterial infection in view of the many clinical symptoms, especially the presence of aphthous and genital ulcerations. An attempt to obtain cutaneous responses to bacterial antigens has been carried out using various vaccines developed from bacteria isolated from the ulcerative lesions and oral cavities of BD patients. BD patients often show intense hypersensitivity to various strains of streptococci, not only by their cutaneous reactions but also by in vitro testing. In this report, we describe our previous studies on the correlation between streptococcal antigens and the pathogenesis of BD and also discuss the recent reports of other authors. The intense hypersensitivity to streptococcal antigens acquired after streptococcal infection is thought to play an important role in the appearance of symptoms in BD patients since the production of pro-inflammatory cytokines by peripheral blood mononuclear cells (PBMC) was enhanced when stimulated with streptococcal antigen in a culture system. Minocycline, an antibiotic to which certain strains of streptococci are sensitive, reduced the frequency of clinical symptoms in BD patients as well as the production of pro-inflammatory cytokines by BD-PBMC stimulated with streptococcal antigen.

Biosynthesis of GA73 methyl ester (GA73-Me), the principal antheridiogen in Lygodium ferns, was investigated. From the methanol extract of prothallia of Lygodium circinnatum, GA25, GA73, GA73-Me, GA88-Me, and a few unknown GA73 derivatives were detected by GC-MS. Because the presence of GA25 suggests that GA24, a direct precursor of GA25, could also be present in L. circinnatum prothallia, we used feeding experiments to investigate the possibility that GA24 is a precursor of GA73-Me. In L. circinnatum prothallia, [2H2]GA24 was converted into [2H2]GA73-Me and a trace amount of [2H2]GA9-Me, whereas [2H3]GA9 was converted into [2H3]GA9-Me and [2H3]monohydroxy-GA9-Me. Because GA73-Me, GA9-Me, and their monohydroxy derivatives had been identified by GC-MS from the culture medium of L. circinnatum prothallia, our results suggest that GA73-Me is biosynthesized from GA24 via GA73, and that neither GA9 nor GA9-Me is a precursor of GA73-Me. Though the possibility had been suggested that GA73-Me is biosynthesized from 9,15-cyclo-GA9 (GA103), [2H2]GA103 was not converted into [2H2]GA73-Me.

An 86-year-old woman was admitted to our hospital with genitourinary bleeding. Abdominal computerized tomography demonstrated a low density mass on the posterior wall of the bladder. Cystoscopy revealed a nonpapillary bladder tumor. A transurethral cold cup biopsy showed lymphocytic infiltration with no malignant cells. She died 2 years after the initial presentation and autopsy revealed malignant lymphoma, diffuse B cell type, small cell type. A cold cup biopsy of the bladder mucosa sometimes fails to detect primary malignant lymphoma of the bladder because of its submucosal location.

An interference-based linamarin sensor is developed. Horseradish peroxidase (HRP) is adsorbed on a pyrolytic graphite (PG) electrode, and then linamarase from cassava is cross-linked with glutaraldehyde on the electrode surface. The prepared bienzyme electrode is poised at -300 mV vs Ag/AgCl for 40 s to reduce dissolved O(2) to H(2)O(2) at the PG surface. The potential is then stepped to 0 mV, at which point the accumulated H(2)O(2) is reduced, though the O(2) reduction does not proceed. Since the H(2)O(2) reduction is catalyzed by HRP, the transient cathodic current is inhibited by cyanide, which is liberated from linamarin by linamarase. Therefore, the transient current is a function of the linamarin concentration. This sensor responds to 1 × 10(-5)-5 × 10(-3) M linamarin and can estimate a linamarin concentration of a cassava extract.

We experienced a case of ganglioneuroma. A 37-year-old man was admitted to our hospital for right flank pain. Computed tomography (CT) showed a low density mass near the upper pole of the right kidney. Selective renal arteriography did not demonstrate any tumor vessels in the right kidney. An endocrinological study revealed high values of noradrenaline and dopamine in urine. Under the preoperative diagnosis of a retroperitoneal tumor or right adrenal tumor, tumor resection was performed. The tumor was found in the retroperitoneal space on surgical exploration. The histopathological diagnosis was ganglioneuroma.

Antheridiogens in two species of Schizaeaceous ferns, Lygodium circinnatum and Lygodium flexuosum, were analyzed by gas chromatography-mass spectrometry. In L. circinnatum, gibberellin A73 (GA73) methyl ester (GA73-Me), which had originally been identified in L. japonicum, was identified as a principal antheridiogen, and the methyl esters of five known GAs (GA9, GA20, GA70, GA88, and 3-epi-GA88) were also identified as minor antheridiogens. In addition, four compounds corresponding to isomers of monohydroxy-GA73-Me were detected. One of these was shown to be 12[beta]-hydroxy-GA73-Me, the parent acid of which has been allocated the GA assignment GA96. The other three compounds, tentatively named X1, X2, and X3, have not been fully characterized. In L. flexuosum, GA73-Me was also identified as a major antheridiogen, with X2 being detected as a minor one. The total antheridium-formation activity in the culture medium of 7-week-old prothallia of L. circinnatum and L. flexuosum was more than 1000 times higher than that of L. japonicum. On the other hand, the response of gametophytes of the former two Lygodium ferns to GA73-Me was more than 100 times lower than that of L. japonicum.

Inexpensive, reagentless, and simple (single-electrode) cyanide biosensors are developed using a pyrolytic graphite (PG) electrode on which horseradish peroxidase (HRP) is adsorbed. The electrode is poised at -300 mV vs Ag/AgCl for 40 s to reduce dissolved O(2) to H(2)O(2) at the PG surface. The generated H(2)O(2) accumulates in the diffusion layer. The potential is then stepped to 0 mV, at which the accumulated H(2)O(2) is reduced, though the O(2) reduction does not proceed. Since the H(2)O(2) reduction is catalyzed by HRP, the transient cathodic current is inhibited by cyanide. Therefore, the transient current is a function of the cyanide concentration. A HRP/PG electrode with saturated HRP coverage is reliable, and it can determine 10(-)(5)-10(-)(3) M cyanide. On the other hand, the electrode with lower HRP coverage is less reliable, though it is so sensitive as to detect 2 × 10(-)(7) M cyanide because the system is under kinetic control.

Recently, it has been noted that radical molecules participate in the pathogenesis of some of the complications associated with long-term dialysis therapy, such as amyloidosis. Since allantoin is produced from uric acid almost exclusively by the hydroxy radical, we investigated the plasma concentrations of allantoin in 71 patients with chronic renal failure who were on maintenance hemodialysis. Our specific objective was to investigate the relation of allantoin to the plasma concentrations of beta2-microglobulin and methylguanidine to evaluate the potential of allantoin as a significant parameter of the radical reaction. Although plasma allantoin was not detected in 15 healthy controls, plasma concentrations of allantoin increased markedly in all of the hemodialysis patients (42.6 +/- 37.7 nmol/mL) and wide variation was observed from 4.3 to 185.2 nmol/mL. In addition, we confirmed significant correlations between plasma concentrations of allantoin and beta2-microglobulin as well as serum concentrations of methylguanidine/creatinine and hyaluronic acid (r - 0.456, p < 0.0005, r = 0.313, p < 0.005, r = 0.368, p < 0.01, respectively). Although evidence for a direct link between plasma allantoin levels and amyloidosis was not obtained, the increase in the plasma concentrations of allantoin suggested its clinical significance as a parameter of the radical reaction that is thought to participate closely in the pathogenesis of dialysis-related amyloidosis. From a practical point of view, the measurement of plasma allantoin can be expected to become a significant method of measuring radical members which may be useful in the prophylaxis of some of the complications induced by free radicals.

The following gibberellins (GAs) and antheridiogens were identified by their mass spectra and Kovats retention indices from combined gas chromatography-mass spectrometry of purified extracts of the prothallia and sporophytes of Anemia phyllitidis, a Schizaeaceous fern: a trace amount of GA9 (4-week-old prothallia); GA9, GA24, GA25, antheridic acid and 3-epi-GA63 (6-week-old prothallia); and GA4, GA9, GA15, GA19, GA20, and GA24 [young sporophytes (younger than one year old) and/or old sporophytes (between one- and two years old). Of these compounds, GA24, GA9, and GA4 were quantified by gas chromatography-selected ion monitoring, using (2)H-GAs as internal standards, and the content of antheridic acid, the principal antheridiogen, was evaluated by a radioimmunoassay which we have developed. The results indicate that endogenous levels of GAs and antheridiogens in prothallia began to increase rapidly between 4 and 6 weeks after sowing, the contents of antheridic acid and GA24, the most abundant GA in 6-week-old prothallia, being 107.4 and 37.9 ng/g fresh weight, respectively. The most abundant GA in the sporophytes was GA9, the content in young and old sporophytes being 15.3 and 7.3 ng/g fresh weight, respectively.

Endogenous gibberellins (GAs) and GA-related compounds at two different developmental stages of apple seeds(Malus domestica cv. McIntosh) were analyzed by gas chromatography-mass spectrometry.From the seeds at ca. 10 and/or 14 weeks after full-bloom, the following GAs and 9,15-cyclogibberellins (9,15-cyclo-GAs) were identified by a comparison of their mass spectra and Kovats retention indices with those of authentic specimens: GA4, GA7, GA9, GA12, GA17, GA19, GA20, GA25, GA34, GA35, GA45,GA53, GA54, GA61, GA62, GA63, GA70, GA73, GA80, GA84, GA88, 3-epi-GA4, 3-epi-GA54, and 3-epi-GA63, 9,15-cyclo-GA9, lβ-hydroxy-9,15-cyclo-GA9, 2β-hydroxy-9,15-cyclo-GA9, 3α-hydroxy-9,15- cyclo-GA9, 3β-hydroxy-9,15-cyclo-GA9, and llβ-hydroxy-9,15-cyclo-GA9. The major components in the seeds at ca. 10 weeks after full-bloom were GA4, GA7, GA9, GA17, GA35, GA54, GA62, GA80, and GA84, and those in the seeds at ca. 14 weeks after full-bloom were GA17, GA25, GA45, GA62, GA63, GA80,GA84, 9,15-cyclo-GA9, and 1β-, 3β-, and 11β-hydroxy-9,15-cyclo-GA9. New GA number (GA103 &#95; 108) have been allocated to 9,15-cyclo-GA9, and 1β-, 2β-, 3β-, 3α-, and 11β-,hydroxy-9,15-cyclo-GA9, respectively (1β- and 3α-hydroxy-9,15-cyclo-GA9 were identified previously as fern anteridiogens, although GA numbers have not previously been assigned to these compounds). This is the first report of the co-occurrence of GA73 and 9,15-cyclo-GAs in higher plants.

A 42-year-old man with giant bronchogenic cyst occupied from the middle and posterior mediastinum was treated surgically. The tumor was stoutly adhered to anterior aspect of the esophagus and membranous portion of the right bronchus. In dissecting the tumor, muscular coat of the esophagus was partly severed, which was repaired interruptedly with absorbable suture material. However, esophageal perforation was detected by barium swallow on the 3rd postoperative day. Conservative therapy included intrathoracic drainage and antibiotics was successfully undertaken. It is imperative to dissect the tumor very carefully if the giant bronchogenic cyst adheres to the esophagus or other surrounding organs.

A 41-year-old female patient with a right retroperitoneal tumor for more than thirty years was referred to our department. Exploration was done through a transperitoneal approach and the tumor was removed. A ganglioneuroma was diagnosed histopathologically. There have been 99 reported cases with retroperitoneal ganglioneuroma including our present case in Japan and we discuss the pathogenesis and treatment of this rare disease.

IA antigens on the cell membrane of inflammatory macrophages and epithelioid cells were investigated with immunoelectron microscopic method during development of granulomas induced by subcutaneous inoculation of 10(7) Mycobacterium lepraemurium into mice with and without hypersensitivity. In C57BL/6N (H-2b) immunogenetic high responder mice 6 weeks after infection majority (87%) of infiltrated cells were IA-positive. Two types of the staining reaction, strong and weak reactivity, were recognized among the positive cells. Strongly IA-positive cells showed lower phagocytosis (0.9/cell section) of mycobacteria than the weakly reacted cells (4.9/cell section). The strongly positive cells underwent morphological differentiation into large epithelioid cells during development of the hypersensitivity-type murine lepromas after 10 or more weeks of infection. Types of granulomas and IA-positive cells in C57BL/6N (nu/+) mice were identical to those found in C57BL/6N. In C57BL/6N (nu/nu) athymic nude mice initial infiltrating cells contained 38% of weakly IA-positive macrophages and a small number (7%) of strongly IA-positive macrophages. But the reactivity was lost later and only 4% of IA-positive cells remained in the granulomas without hypersensitivity. CBA/J (H-2k) low responder mice did not show IA-positive cells in either initial or late stage during the development of nonhypersensitivity-type murine lepromas. We suggest that the presence of IA-positive cells, particularly IA-positive epithelioid cells, in the lesions modulates the course of granulomatous tissue reaction in murine lepromas.

A 76-year-old man, who had undergone surgery for lung cancer, was referred to our department for further examination for left renal mass. Exploration was done through a transperitoneal approach and the left kidney was removed. A metastatic renal tumor originating from a pulmonary squamous cell carcinoma was diagnosed histologically. There have been 73 reported cases with metastatic renal tumor in Japanese literature and we reviewed the pathogenesis and treatment of this rare entity.

The role of Ca2+ in stimulation of the malate-aspartate shuttle by norepinephrine and vasopressin was studied in perfused rat liver. Shuttle capacity was indexed by measuring the changes in both the rate of production of glucose from sorbitol and the ratio of lactate to pyruvate during the oxidation of ethanol. (T. Sugano et al. (1986) Amer. J. Physiol. 251, E385-E392). Asparagine (0.5 mM), but not alanine (0.5 mM) decreased the ethanol-induced responses. Norepinephrine and vasopressin had no effect on the ethanol-induced responses when the liver was perfused with sorbitol or glycerol. In the presence of 0.25 mM alanine, norepinephrine, vasopressin, and A23187 decreased the ethanol-induced responses that occurred with the increase of flux of Ca2+. In liver perfused with Ca2+-free medium, asparagine also decreased the ethanol-induced responses, but norepinephrine and vasopressin had no effect. Aminooxyacetate inhibited the effects of norepinephrine, A23187, and asparagine. Regardless of the presence or absence of perfusate Ca2+, the combination of glucagon and alanine had no effect on the ethanol-induced responses. Norepinephrine caused a decrease in levels of alpha-ketoglutarate, aspartate, and glutamate in hepatocytes incubated with Ca2+. The present data suggest that the redistribution of cellular Ca2+ may activate the efflux of aspartate from mitochondria in rat liver, resulting in an increase in the capacity of the malate-aspartate shuttle.

A strain of Mycoplasma hominis lacking a major membrane protein of 120 kDa was isolated from a Vero cell culture. This strain showed very slow growth rate and formed nipple-less colonies on agar medium.