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The Arabidopsis protein COP1, encoded by the constitutive photomorphogenic locus 1, is an essential regulatory molecule that plays a role in the repression of photomorphogenic development in darkness and in the ability of light-grown plants to respond to photoperiod, end-of-day far-red treatment, and ratio of red/far-red light. The COP1 protein contains three recognizable structural domains: starting from the N terminus, they are the zinc binding motif, the putative coiled-coil region, and the domain with multiple WD-40 repeats homologous to the beta subunit of trimeric G-proteins (G beta). To understand the functional implications of these structural motifs, 17 recessive mutations of the COP1 gene have been isolated based on their constitutive photomorphogenic seedling development in darkness. These mutations define three phenotypic classes: weak, strong, and lethal. The mutations that fall into the lethal class are possible null mutations of COP1. Molecular analysis of the nine mutant alleles that accumulated mutated forms of COP1 protein revealed that disruption of the G beta-protein homology domain or removal of the very C-terminal 56 amino acids are both deleterious to COP1 function. In-frame deletions or insertions of short amino acid stretches between the putative coiled-coil and G beta-protein homology domains strongly compromised COP1 function. However, a mutation resulting in a COP1 protein with only the N-terminal 282 amino acids, including both the zinc binding and the coiled-coil domains, produced a weak phenotypic defect. These results indicated that the N-terminal half of COP1 alone retains some activity and a disrupted C-terminal domain masks this remaining activity.

Sarcomatoid carcinoma is a rare variant of malignant tumor arising from the urinary tract. This tumor had been termed carcinosarcoma because of its carcinomatous and sarcomatous components. There is still some confusion in the terminology between true carcinosarcoma and sarcomatoid carcinoma; however, the latter is now regarded as primarily a malignant epithelial tumor with pseudosarcomatous transformation. Four cases of sarcomatoid carcinoma arising from the urinary tract are reported. The patients were a 77 year old female, and three males aged 62, 69 and 80 years. All but the eldest patient complained of gross hematuria. Surgical removal was performed in the younger three cases, and an autopsy was done in the remaining case. All the tumors were macroscopically polypoid. Histopathologic examination revealed fasciculated spindle-cell tumors with myxoid stroma or malignant fibrous histiocytoma-like spindle cell tumors. The epithelial nature was proven in these sarcomatous cells by immunohistochemical and/or electron-microscopic examinations. Only a small amount of squamous cell carcinoma components was also evident in the latter three cases. Although the younger three patients were alive at 44, 23 and 39 months' follow-up, respectively, constant careful monitoring is recommended.

The transition from vegetative growth to flowering occurs at the shoot apical meristem (Simpson et al. 1999; Parcy 2005). Floral induction causes an apical meristem to produce flowers, which consist of a complex array of specialized structures (Zeevaart 1976; Bernier 1998). Flowering is regulated by signals from endogenous and external sources. Endogenous signals include circadian rhythms, developmental stage and hormones, while external signals comprise day length and temperature.

The Arabidopsis thaliana PROTODERMAL FACTOR1 (PDF1) gene encoding a putative extracellular proline-rich protein is exclusively expressed in the L1 layer of shoot apices and the protoderm of organ primordia. In order to identify essential cis-regulatory sequences required for the L1 layer-specific expression, a series of 5' deletions of the PDF1 promoter were fused to the beta-glucronidase (GUS) gene and introduced into Arabidopsis plants. Our analysis revealed that the minimum region necessary to confer L1-specific expression of PDF1 is confined within a 260-bp fragment upstream of the transcription start site. We identified an 8-bp motif in this region that is conserved between promoter regions of all the L1-specific genes so far cloned, and we designated it the L1 box. Electrophoretic mobility shift assays demonstrated that the L1-specific homeodomain protein ATML1 can bind to the L1 box sequence in vitro. The GUS expression in transgenic plants disappeared when a mutation that abolishes binding of ATML1 was introduced into the PDF1 l1 box sequence of the construct. These results suggest that the L1 box plays a crucial role in the regulation of PDF1 expression in L1 cells and that ATML1 could cooperate to drive L1-specific expression.

DCECP63 is a carrot embryogenic cell protein. To perform its genetic analysis, we isolated an Arabidopsis thaliana (At) ECP63 cDNA. The cDNA encoded a polypeptide of 449 amino acids (aa). Its deduced aa sequence showed extensive similarity with DCECP63, DCDC8 and BPBP8 which are embryonic proteins isolated from carrot and birch, respectively. The aa sequence contained eight well-conserved tyrosine phosphorylation sites and 16 repeats of 11 aa. Southern analysis showed that the AtECP63 gene might belong to a small multigene family. The AtECP63 transcripts accumulated specifically in mature seeds, and exogenous abscisic acid (ABA) induced its expression in immature siliques, but not in vegetative tissues. These results suggested that the AtECP63 gene encoding a putative phosphotyrosine protein belonging to late embryogenesis abundant (LEA) protein in group 3 might be involved in maturation and desiccation tolerance of seeds. Restriction fragment length polymorphism (RFLP) and cleaved amplified polymorphic sequence (CAPS) mapping showed that AtECP63 gene was present in the South part of chromosome 4.

We analyzed the 99mTc-DTPA renogram with and without diuresis to predict the possibility of stone discharge on the outpatient basis by renogram patterns. Between October, 1993 and December, 1995, 99mTc-DTPA renography was performed in 79 patients with a single ureteral stone. The 99mTc-DTPA renogram pattern was classified into the three types of normal function, obstruction and lower function patterns and the complete stone discharge rate was 93, 63 and 25%, respectively. In addition, diuretic renography using Furosemide was performed in patients with an obstruction pattern and the three renogram patterns of return to the normal curve, a diuretic response and no response were obtained; the complete stone discharge rate was 44, 65.3 and 93%, respectively. From this study, patients with a single ureteral stone with a normal pattern on the regular DTPA renogram and patients with no response pattern on the diuretic renogram, even if in such patients an obstructive pattern was seen on the regular DTPA renogram, seem to be a good candidate for obtaining a high rate of a stone discharge with extracorporeal shock wave lithotripsy (ESWL) treatment in the outpatients basis.

From April 1994 to February 1995, we studied prostate cancer screening at Shakaihoken Hazu Hospital and Health Care Center. Among 1,838 men, over 55, who had taken the total health care check, 857 applicants had had prostate cancer screening, using serum PSA (Delia kit; normal range11.7 ng/ml) and AUA voiding symptom score and bothersome score. According to the serum PSA level, they were divided into 4 groups, 11 with 8 ng/ml or more indicating requirement of further examination, 29 with 4-8 ng/ml indicating requirement of observation, 100 with less than 4 ng/ml and AUA score (approximately more than 10) indicating voiding disturbance and the other 717 were normal; Of 11 further examined cases, 6 applicants underwent random biopsy. We found 3 with prostate cancer (0.35%).

Male-associated DNA sequences were analyzed in a dioecious plant, Cannabis sativa L. (family: Moraceae), which is known to have sex chromosomes. DNA was isolated from male and female plants and subjected to random amplification of polymorphic DNA. Two out of 15 primers yielded fragments of 500 and 730 bp which were detected in all male plants but not in any of the female plants tested. These two DNA fragments were cloned and used as probes in gel blot analysis of genomic DNA. When the male and female DNAs were allowed to hybridize with the 500-bp probe, no differences in patterns were observed between male and female plants. By contrast, when these DNAs were allowed to hybridize with the 730-bp probe, much more intense bands specific to male plants were detected, in addition to less intense bands that were common to both sexes. The 730-bp DNA fragment was named MADC1 (male-associated DNA sequence in Cannabis sativa). The sequence of MADC1 did not include a long open reading frame and it exhibited no significant similarity to previously reported sequences.

Herein, we report two cases of squamous cell carcinoma of the ureter. The first case was in a 56-year-old-male. Total cystectomy and ileal conduit were performed because of bladder tumor suspected to be accompanied by carcinoma in situ and atrophic urinary bladder induced by chronic cystitis in December, 1993. Pathological examination revealed transitional cell carcinoma (TCC)squamous cell carcinoma (SCC), G2G1, INF beta, pT1, 1y1, v1. He complained of back pain under medical observation in December, 1994. Left hydronephrosis was found and antegrade pyelography showed leakage from the left pelvic ureteral junction. Urinary cytology revealed class V and suggested TCC. He received left nephroureterectomy, and pathohistological examination of resected specimen revealed SCC, INF gamma, pT3, pRo, pLx, pVx, pNo, pMo. CABO chemotherapy (cisplatin, methotrexate, bleomycin, vincristine) was performed postoperatively. The second case was in a 61-year-old female. She complained of macrohematuria in the course of observation of pyelonephritis. Drip infusion pyelography showed right hydronephrosis and retrograde ureterogram revealed stenosis of the right lower ureter. Urinary cytology revealed class V. Nephroureterectomy and bladder cuff were performed. The tumor was histologically diagnosed as SCCTCC, INF beta, pT3, pRo, pLo, pVo, pNo, pMo. Postoperatively, CABO chemotherapy was performed. So far, no recurrence has been observed. Fifty five cases of squamous cell carcinoma of ureter were collected from the Japanese literatures including our cases.

We report one case of Paneth cell-like change of prostate cancer. An 81-year-old male reported to our hospital with the chief complaint of urinary retention. The serum concentration of prostate specific antigen, 168 ng/ml, was high, and digital examination was performed. Because we suspected prostate cancer, needle biopsy of the prostate was performed. Histological examination revealed moderately-poorly differentiated adenocarcinoma. Computed tomography revealed no invasion to the prostatic wall and no metastasis to the lymph node. 99mTc-HMDP revealed no bone metastasis. We chose intravenous hormonotherapy (fosfestrol 500 mg/day, 20 days), but urinary retention did not improve. Therefore, we performed transurethral prostatectomy to improve the symptoms. Histological examination of the removed specimen revealed moderately differentiated adenocarcinoma with Paneth cell-like change. Immunochemical and histochemical stains were positive for Grimelius, serotonin and prostatic acid phosphatase and negative for periodic acid-Schiff reaction, lysozyme, neuron specific enolase, prostate specific antigen, alpha-1-antitrypsin and IgA.

The most readily oxidizable sites in one-electron oxidation of duplex DNA are shown to be the guanine (G) residues located 5' to guanine. Based on experimental results and ab initio calculations, G-G stacking rule has been proposed. This rule indicates the most electron-rich sites contained in B DNA and is very useful for understanding CT interactions between electron-accepting molecules and DNA.

2'-Bromo-substituted cyclopentane-type carbocyclic nucleoside (7a, 7b) was synthesized from sodium cyclopentadienylide. Subsequently, 7a was incorporated into DNA oligomer by means of a conventional phosphoramidite method. Melting-temperature, CD spectra and chemical reaction of such carbocyclic oligonucleotide are described.

Production of a functional beta-glucuronidase (GUS) protein was induced by exposure of exponentially growing yeast cells to heat shock after transformation of the GUS gene under the control of the promoter of the heat-shock gene, HSP18.2, from Arabidopsis. Yeast cyr and bcy mutations appeared to have essentially no effect.

To determine the current status of urolithiasis in 1988, in comparison with that in 1985, we analyzed the 1937 patients of urolithiasis at 17 departments of urology in Mie Prefecture and 2 departments of urology in Wakayama Prefecture. The ratio of male to female patients was 2.6 to 1.0. Geographically, the number of urolithiasis patients was most frequently distributed in Matsusaka City. The frequency of urolithiasis in the urban area was almost the same as that in the rural area. Most of the stones (96.3%) were in the upper urinary tract. The frequency of lower urinary tract calculi tended to be high in southern Mie Prefecture. The ratio of the upper urinary tract calculi to the lower urinary tract calculi in the urban area was higher than in the rural area. The age distribution in males was in the forties, while that in females was in the fifties. The average age was 46.4 years old. The surgical treatment was performed in 671 patients (34.6%) and the extracorporeal shock wave lithotripsy (ESWL) was the most frequent mode of treatment (85.0%), followed by cysto-lithotripsy (4.2%) and percutaneous nephro-uretero-lithotripsy (2.4%). The most frequent component of the urinary tract calculi was calcium oxalate and/or calcium phosphate (81.7%). The stone patient increased in the number during the summer season (July, August and September). In conclusion, in 1988 when the ESWL treatment started in Mie Prefecture, the epidemiologic features of urolithiasis was characterized as follows: the number of patients increased and the broad application of the ESWL treatment resulted in the decreased number of patients with spontaneous discharge and the increased number of patients with recurrent stones and with bilateral or multiple complex stones.

Two mutants of Escherichia coli strictly deficient in uridine-diphosphoglucose pyrophosphorylase activity (galU) were found to have very small numbers of flagellar filaments and hooks. In these mutants, both the rate of flagellin (flagellar protein) synthesis and the amount of messenger ribonucleic acid specific for flagellin were considerably lower than in the parental strains. Motile revertants from the galU mutants were isolated and were found to carry a suppressor mutation, which was mapped in the flaH cistron. These strains formed swarms under conditions of catabolite repression; their intracellular concentration of cyclic adenosine 5'-monophosphate was the same as that in the parental strains. These results suggest that the outer membrane affects flagellar formation through the flaH gene product.

Of the more than 30 genes required for flagellar function, 6 are located between pyrC and ptsG on the Escherichia coli genetic man. This cluster of genes is called flagellar region I. Four-point transductional crosses were used to establish the position and order of the region I flagellar genes with respect to the outside markers ptsG and pyrC. Bacteriophage lambda-E. coli hybrids that contained most of the genes necessary for flagellar formation were constructed. The properties of specific hybrids that carried the region I fla genes were examined by genetic complementation and by measuring the capacity of the hybrids to direct the synthesis of specific polypeptides. The results of these tests with lambda hybrids and with a series of deletion mutations derived from the lambda hybrids demonstrated the existence of at least six flagellar-specific cistrons. These directed the synthesis of polypeptides with the following apparent molecular weights: flaV, 11,000; flaK, 42,000; flaL, 30,000 and 27,000; flaM, 38,000; flS, 60,000; and flaT, 35,000. Plasmid ColE1-E. coli hybrids with region I flagellar genes were also used to program the synthesis of polypeptides in minicell-producing strains. The polypeptides synthesized in these experiments were identical to polypeptides of the hook-basal body structure and helped to confirm the assignment of genes to specific polypeptides. The synthesis of all of these polypeptides was regulated by the same mechanism that regulates the synthesis of other flagellar-related structural components.

Previous studies showed that the structural gene for the flagellar hook subunit protein (molecular weight 42,000) was one of a group of flagellar genes located on the Escherichia coli genome near pyrC. Several lines of evidence indicate that the flaK gene is the structural gene for the hook subunit protein. Fla+ strains that were insensitive to chi infection could be isolated as revertants of an FlaK- amber mutant strain but from no other Fla- strain. The hook subunit proteins isolated from such chi-sensitive revertants of the FlaK- strain were shown to be antigenically and electrophoretically different from the hook protein isolated from the wild-type strain. Thus, reversion of a mutation in the flaK gene resulted in alteration of the structure of the hook protein. Furthermore, in programming experiments with hybrid lambda containing flagellar genes, lambdafla with flaK genetic activity programmed the synthesis of a 42,000-molecular weight protein, whereas lambdafla without flaK genetic activity did not.

The Escherichia coli operon designated flaA contains seven flagellar genes; among them are two switch protein genes whose products are believed to interface with the motility and chemotaxis machinery of the cell. Complementation analysis using several plasmids carrying different portions of the flaA operon and analysis of expression of these plasmids in minicells allowed the identification of two flagellar gene products. The MotD (now called FliN) protein, a flagellar switch protein, was determined to have an apparent molecular weight of 16,000, and the FlaAI (FliL) protein, encoded by a previously unidentified gene, had an apparent molecular weight of 17,000. DNA sequence analysis of the motD gene revealed an open reading frame of 414 base pairs. There were two possible initiation codons (ATG) for motD translation, the first of which overlapped with the termination codon of the upstream gene, flaAII (fliN). The wild-type flaAI gene on the chromosome was replaced with a flaAI gene mutated in vitro. Loss of the flaAI gene product resulted in a nonmotile and nonflagellated phenotype. The subcellular location for both the MotD and FlaAI proteins was determined; the FlaAI protein partitioned exclusively in the insoluble fraction of a whole minicell sonic extract, whereas the MotD protein remained in both the soluble and insoluble fractions. In addition, we subcloned a 2.2-kilobase-pair DNA fragment capable of complementing the remaining four genes of the flaA operon (flbD [fliO], flaR [fliP], flaQ [fliQ], and flaP [fliR]).

A series of fusions of flagellar genes to the lacZ gene was generated by insertion of Mu dII301 (Apr lac) bacteriophage into the genome of Escherichia coli. The beta-galactosidase activity in each resulting mutant was measured, and the location of the activity in the membrane, periplasmic, or cytoplasmic fraction of the cell was determined. There were three classes of mutants: those which had beta-galactosidase activity mainly in the membrane fraction, those which had it distributed in the soluble and membrane fractions, and those which had it in the cytoplasmic fraction only. The last, soluble-fraction-only, class was predominant in fla-lac gene fusions. In particular, the following mutants were shown to have beta-galactosidase activity in the membrane fractions: on the inner membrane, mutants with flaB fusions, and on the inner and outer membranes, mutants with flaA4850, flaM, and flaU4849 fusions. These results suggest that fla-lacZ gene fusions produce proteins which are able to detect the signals of the leader sequence and the membrane-anchoring region of the flagellar system.

Flagellar mutants in Escherichia coli region I were obtained by selection for resistance to the flagellotropic phage chi. F' elements carrying this region of the E. coli genome were then constructed. Stable merodiploid strains with a flagellar defect on the exogenate and another on the endogenote were prepared. These merodiploids yielded information on the complementation behavior of mutations in this region. Region I was shown to include at least six cistrons, flaV, flaK, flaL, flaM, flaS, and flaT. Mu-induced and deletion fla mutants were also isolated. By using these mutant strains, the transcriptional order was shown to be flaV-flaK-flaL-flaM-flaS-flaT. The definition of region I fla genes and their transcriptional relationships were confirmed by genetic tests with hybrid A phage carrying fla genes in this region.

In order to elucidate the clinical implications of cortisol levels in pregnancy, maternal and fetal serum levels of cortisol during pregnancy were determined. The maternal serum level of cortisol gradually increased with advancing gestational age, and it reached a 3 to 4 times higher level in the late stage of pregnancy than that in non-pregnant state. The maternal serum level of cortisol during delivery increased with the length of the period of labor, and immediately after the delivery it reached the peak value of 60-80 micrograms/dl. According to the mode of delivery, the cortisol level increased with the accumulation of stress on the maternal side. The value was highest in the case of vacuum extraction and lowest in elective cesarean section. The cortisol level in umbilical arterial blood in normal delivery was not significantly elevated in comparison with that in vacuum extraction, oxytocin induced delivery and elective cesarean section. The cortisol level in umbilical blood of anencephaly did not differ from that of normal neonate, indicating that cortisol in umbilical blood may originate mainly on the maternal side. In the postpartum period, the serum cortisol concentration declined to the level observed in the late stage of pregnancy after 2 days, and further decreased to the level in the non-pregnant state after 6 days. The present studies suggest that the increase in the serum concentration of cortisol at delivery is a result of maternal stresses in labor, and that direct involvement of cortisol in the controlling mechanism of puerperium is unlikely. The increased function of the adrenal cortex in the late stage of pregnancy and at delivery may stimulate fetal pulmonary maturity and provide maternal and fetal adaptability to stresses in intrapartum bleeding and shock during labor. It is essential to understand pituitary-adrenal cortex functions in prenatal and fetal care in pregnancy.

We report a 3-month-old boy who presented with acute right scrotal swelling that was diagnosed to have epididymitis at surgical exploration. Urine culture and urinalysis were negative and IVP revealed no abnormality. White blood count and symptom were improved with chemotherapy. Acute epididymitis in infancy is very rare and this case seems to be the 16th case in Japan.

Specialized transducing lambda phages carrying the region III flagellar genes (fla) of Escherichia coli K-12 were isolated by a new method. A strain carrying both a cryptic lambda prophage near the his genes and a deletion of the attlambda gene was used as a starting strain. The lysogen of lambdacI857pga18-bio69 was isolated in which the prophage was integrated within the lambda cryptic genes by means of recombination with the residual lambda DNA. The strains with deletions starting within the prophage and ending in these fla genes were selected from among the heat-resistant survivors of the lysogen. They were then infected with heat-inducible and lysis-defective lambda phages and, thus, specialized transducing phage lines for hag and fla were obtained. High-frequency transfer lines of rare phages carrying the fla genes were isolated by inducing a strain carrying a heat-inducible lambda prophage near the his genes and selecting by transduction of a fla deletion strain. Preliminary characterization of these transducing phages is also reported.

In order to study the secretory mechanism of human placental lactogen (hPL:hCS) from trophoblastic tissues, tissue culture and new placental perifusion systems were developed and used to clarify the effect of various substances on the secretion of hPL. These substances were (1), metal ions([Ca2+], [K+], [mg2+], [Na+]); (2) growth hormone and prolactin releasing or inhibiting factors (arginine, TRH, somatostatin, dopamine); (3) LH-RH, dibutyryl cyclic-AMP which stimulates hCG secretion; (4) prostaglandins F2 alpha and E2, bradykinin; (5) EGF and insulin which have the receptors in the placenta; (6) glucose. It was found that most of the substances examined had no effect on the secretion of hPL, except dopamine and glucose. The effect of dopamine in the tissue culture system is dose-dependent. At high concentrations dopamine slightly inhibited hPL secretion(5mM: 38.6 +/- 15.0 and 10mM; 35.7 +/- 19.0 micrograms/g wet tissue) compared with the control (63.2 +/- 29.8 microgram/g wet tissue). However, these effects may be due to the deviation of pH in the medium from the direct addition of dopamine hydrochloride. At a low concentration(1mM) it was observed to have a rather stimulatory effect (125.7 +/- 18.0 micrograms/g wet tissue, p less than 0.05), but in the perifusion system, this effect could not be observed. The addition of glucose in the perifusion system gave a slightly higher hPL secretion than that of the control. Perhaps this resulted from increased cell activity rather than a stimulatory effect. an incorporation experiment of [3H] leucine was also carried out to study the biosynthesis of hPL. Newly synthesized ([3H]-labelled) hPL was secreted into the medium within two hours. Furthermore, a labelled larger molecular weight substance together with the tritiated hPL was also observed on a Sephadex G-100 gel chromatography. These labelled substances were immunoprecipitable using an anti-hPL serum, indicating that the substances contain the same immunological determinants. This result indicates that the larger molecular substance may represent the biosynthetic precursor or the aggregate of hPL. These data indicate that the secretion of hPL has a unique mechanism, different from GH and PRL, and may be self regulated.

Previous studies have defined 29 genes necessary for synthesis of the Escherichia coli flagellar apparatus. This study analyzed the transcriptional control of flagellar genes, using Mu d (Apr lac) phage to generate flagellar mutants by insertion. These mutants contained operon fusions of flagellar genes to the lac genes of the Mu d phage and allowed the measurement of flagellar operon expression by detection of beta-galactosidase activity. These fusion mutants expressed the enzyme activity constitutively, and an autogenous regulation mechanism was not revealed. Lambda transducing phages carrying these chromosomal fla-lac fusions were also isolated and used to examine the effect of different fla mutations on expression of each flagellar operon. The results showed that flagellar operons are divided into six classes; (class 1) the flbB operon, which controls all of the other flagellar operons; (class 2) the flaU and flbC operons, which are controlled by the flbB operon gene products and are not required for the expression of other Fla operons; (class 3) the flbA, flaG, flaD, flaN, flaB, and flaA operons, which are under flbB operon control and are required for the expression of other fla operons; (class4) the flaZ operon, which is controlled by the gene products of the group 1 and 3 operons and is required for hag transcription; (class 5) the mocha and flaS operons, which are controlled by the gene products of the group 1 and 3 operons; and (class 6) the hag operon. These results are discussed with respect to the possible assembly sequence of the fla gene products.

The diagnosis and management of 6 patients with spontaneous non-traumatic urinary extravasation are reported. It is important to distinguish extravasation of the fornical backflow type from that owing to frank rupture of the diseased renal pelvis. In Japan, 26 cases of spontaneous peripelvic extravasation and 29 cases of spontaneous pelvic rupture have been reported. Most of them were caused by calculous ureteral obstruction. Many of the cases of spontaneous peripelvic extravasation could be managed conservatively, and many of the cases of spontaneous pelvic rupture often required surgical treatment due to the patient's clinical condition, the persistence of obstruction or extravasation, or the presence of complication of extravasation such as urinoma or abscess. All of our 6 cases were managed conservatively and had no complications.

We performed a survey on the iodide test and its adverse effects on a series of 829 DIP patients who had taken the test at our clinic between November, 1984 and August, 1985. Adverse effects were seen in four out of nineteen positive cases of the iodide test. Among the negative 810 cases, 79 were found to present with adverse effects though not statistically significant. Twenty-six out of 131 cases of positive past history of allergy showed a positive reaction whereas 55 out of 698 negative cases showed adverse effects, with a statistical significance at p less than 0.01. The adverse reaction occurred immediately after injection in the majority of cases and in 85%, the reaction was seen after injection of half of the drug. Symptoms also appeared thirty minutes after injection in some cases. Immunological evaluation of the allergy was done in fifteen positive cases. Four cases showed an increase in peripheral eosinophils, three cases showed increased IgE(RIST) and three cases manifested with a decreased CH 50. Urografin-induced lymphocyte blastogenesis (DLST) was positive in six cases. Four cases showed abnormality in more than two tested parameters, and three cases showed complete normal values in all tests. The adverse effects due to contrast medium are considered to occur not as one type or allergic reaction, but to involve other factors as delayed-type reaction which was also seen in 40% of the patients. Close observation of patients after examination with contrast medium is thus mandatory.(ABSTRACT TRUNCATED AT 250 WORDS)

The production of hook protein and flagellin in 29 Fla- mutants of Escherichia coli K-12 was determined by the complement fixation assay. Six mutants produced hook protein, and four of them also produced flagellin. A flaE mutation was introduced into these fla mutants carrying the hook structure. All of these mutants made polyhooks and were used as hosts for a newly isolated host-range mutant of chi phage that has a high affinity for the hook structure. All except one mutant produced significant amounts of progeny phages. A flaD flaE double mutant was that exception which did not yield significant amounts of progeny by the phage propagation method. All of the flaE double mutants produced comparable amounts of polyhooks, and no qualitative difference was detected between chi-sensitive and chi-insensitive mutants by the complement fixation assay. Accordingly, it was thought that the polyhook of the flaD flaE mutant had a mechanical defect for chi phage infection. This assumption was confirmed by tethered-cell experiments; the flaD flaE mutant did not rotate. These results are well explained by a proposed regulation pathway of flagellar genes. flaE mutants can express other genes which govern the final step of the flagellar morphogenesis, whereas flaD mutants cannot rotate, possibly because the mocha operon is not expressed. The results obtained in E. coli were also found to be applicable to Salmonella typhimurium.

flaFIX, the structural gene for the periplasmic P ring of the flagellar basal body of Salmonella typhimurium, was cloned. Two gene products with apparent molecular weights of 38,000 and 40,000 were identified by minicell analysis. Data from pulse-chase and membrane fractionation experiments and data on the inhibitory effect of the proton ionophore carbonyl cyanide m-chlorophenylhydrazone all indicated that the 40-kilodalton protein was a precursor form which, after export across the cytoplasmic membrane accompanied by cleavage of a signal peptide, gave rise to the mature protein in the periplasm. The N-terminal amino acid sequence of the FlaFIX protein, predicted from the DNA sequence, conformed well to known signal peptide sequences. The results indicate that the P-ring protein of the basal body (unlike flagellin and possible some other external flagellar components) crosses the cytoplasmic membrane in a conventional signal peptide-dependent manner.

Autoregulation of the expression of flagellar genes was investigated by the technique of operon fusion. The results suggested that the flaU gene is a repressor and the flaD gene is an activator of transcription of the hag, flaS, and Mocha operons. The action of the putative flaU repressor appears to be masked by its interaction with other flagellar proteins during assembly; thus, repression is apparent only when the interacting proteins are absent. This hypothesis is supported by the phenotype of an unusual flaU mutant, which represses even though it is unable to promote flagellar assembly. Presumably, the mutant synthesizes a repressor whose activity is no longer masked by interaction with other flagellar proteins.

Previous studies have defined 28 genes necessary for the synthesis of the flagellar apparatus of Escherichia coli K-12. This study analyzed the influence of the flagellar genes on the expression of the hag gene (structural gene for flagellin). To this end, a hag::Mu d(Apr lac) mutant which had the lac genes fused to the promoter of the hag gene was constructed. This allowed the measurement of hag gene expression by detection of beta-galactosidase activity. The following observations were made. (i) The hag gene was expressed constitutively in Fla+ cells. (ii) hag gene expression was positively regulated by flaA, FLAB, flaC, flaD, flaE, flaG, flaH, flaI, flaK, flaL, flaM, flaN, flaO, flaP, flaQ, flaR, flaV, flaW, flaX, flaY, flaZ, flbA, and flbB genes.hag-lac expression was not observed in strains with these fla mutations. (iii) The hag gene was expressed in mutants with flaS, flaT, flaU, and flbC defects. Therefore, these genes were not involved in regulation of hag gene transcription.

Evoked response of bulbocavernosus reflex was taken in 8 normal control individuals and in 15 abnormal voiding patients (8 benign prostatic hyperplasia, 4 diabetic neuropathy, 3 supranuclear neurogenic bladder). the latency in control group showed 31.5 +/- 5.0 msec duration after penile stimulation and 65.4 +/- 11.0 msec duration after posterior urethral stimulation. The latency in diabetic neuropathy group was obviously prolonged but not that in the supranuclear neurogenic bladder group. This modern neurophysiological electrostimulation method provides simple and accurate information of bulbocavernosus reflex.