No evidence for DNA and early cytogenetic damage in bystander cells after heavy-ion microirradiation at two facilities.

The occurrence of bystander effects has challenged the evaluation of risk for heavy ions, mainly in the context of space exploration and the increasing application of carbon ions in radiotherapy. In the present study, we addressed whether heavy-ion-induced DNA and cytogenetic damage is detectable in bystander cells. The formation of gamma-H2AX foci, sister chromatid exchanges and micronuclei were used as markers of damage to DNA. Normal human fibroblasts were exposed to low fluences of carbon and uranium ions, and alternatively single cells were targeted with heavy ions using the GSI microbeam. We did not observe a significant increase in the bystander formation of gamma-H2AX foci, sister chromatid exchanges or micronuclei. In addition, we performed for the first time parallel experiments at two microbeam facilities (GSI, JAEA) using the same cell line, culture conditions and irradiation protocols. No significant enhancement of the micronucleus frequencies in bystander cells was detected after targeted carbon-ion irradiation, confirming the results. Details regarding the history, culture conditions or support of the cells might be affecting the detection of bystander effects. On the other hand, the potential X-ray- and heavy-ion-induced bystander effects investigated herein clearly do not exceed the experimental error and thus are either lacking or are less pronounced than the effects reported in the literature for similar end points after alpha-particle and X-ray exposure.

4-N-hydroxy-L-2,4-diaminobutyric acid. A strong inhibitor of glutamine synthetase.

Analogues of glutamic acid were synthesized and evaluated for their inhibitory activity toward glutamine synthetase (EC 6.3.1.2; GS). The title compound, 4-N-hydroxy-L-2,4-diaminobutyric acid (NH-DABA), showed a potent inhibitory activity against GS from both sheep brain and soybean. The inhibition is competitive with respect to glutamic acid and the Ki values of sheep brain GS and soybean GS for NH-DABA are 0.007 mmol and 0.021 mmol, respectively. The activity of inhibition is comparable to those of L-methionine sulfoximine and 2-amino-4-(hydroxymethyl-phosphinyl)butyric acid (phosphinothricin).

Prognostic significance of ERBB3 overexpression in oral squamous cell carcinoma.

Immunohistochemical analysis of erbB3, as the third member of epidermal growth factor receptor gene family, was performed on 41 cases of oral squamous cell carcinoma, correlating the staining pattern with clinical outcome. High expression of erbB3 protein (ERBB3) was significantly associated with lymph node metastasis (P < 0.05), survival rate (P < 0.05) and mode of invasion (P < 0.01) in this series. These results demonstrated that ERBB3 expression may be helpful in identifying those oral squamous cell carcinomas with higher malignant potential.

Mechanisms of gastric juice-induced hyperpermeability of the cultured human tracheal epithelium.

PURPOSE: The respiratory aspiration of the stomach contents causes severe lung damage called aspiration pneumonia. The present study was undertaken to elucidate whether mucosal exposure of gastric juice causes hyperpermeability of the human airway epithelium and to determine the mechanisms responsible for gastric juice-induced airway epithelial damage. MATERIALS AND METHODS: Gastric juice was collected from 46 normal adults via gastroscope and samples were analyzed for pH, osmolarity, and concentration of pepsin and trypsin. Tracheal surface epithelial cells were obtained from 16 autopsies, cultured onto porous filters, and mounted in the Ussing chamber. Electrical conductance (G) was measured before and after exposure of cells to gastric juice or Krebs-Henseleit solution with pH at 1.8, 2.8, 4.0, or 7.4 in the presence or absence of pepsin. D-[3H] mannitol flux study across the epithelial layer and histologic observations using an inverted microscope were also performed after exposure of cells to gastric juice. RESULTS: Exposure of cultured human tracheal epithelium to gastric juice caused increases in G in a time- and pH-dependent fashion. A pepsin inhibitor (pepstatin A) inhibited gastric juice-induced increases in G at a pH of 2.8, and the addition of pepsin augmented increases in G induced by the Krebs-Henseleit solution at a pH of 1.8 and 2.8. Lowering the osmolarity of the solution to levels similar to gastric juice also potentiated increases in G induced by acid and pepsin. Gastric juice caused increases in D-[3H] mannitol flux across the epithelial layer bidirectionally, and microscopic observation revealed separation of the intercellular space and cell detachment from culture vessels after exposure of cells to gastric juice. CONCLUSION: Gastric juice causes hyperpermeability across human airway epithelium probably through the additive effects of gastric acid, pepsin activity, and lower osmolarity.

Overexpression of Medaka (Oryzias latipes) photolyase gene in Medaka cultured cells and early embryos.

To study the role and the regulation of the photolyase gene in the Medaka (small teleost), we constructed a eukaryotic expression plasmid of the Medaka photolyase gene and introduced it into Medaka cells in vivo and in vivo. The expression plasmid contains a cytomegalovirus enhancer and a thymidine kinase promoter to overexpress the photolyase gene of the Medaka. First, we transfected this construct into cultured Medaka cells and established several lines of transfectant. Every transfectant showed enhanced ability of pyrimidine dimer repair in the presence of fluorescent light. In the transfectant that showed the most enhanced ability of photorepair, the augmented transcription of photolyase gene was observed compared with that of progenitor OL32 cells. In this transfectant, we also observed an enhanced rate of UV survival with 20 min of fluorescent light treatment after irradiation with a 400 J/m2 UV sunlamp. Next, the expression construct was microinjected into the embryos of the Medaka at the one cell stage. Compared with the nontreated counterparts, the overexpression of a photolyase gene was detected in the microinjected embryos, but we failed to detect a significant increase in photo-reactivability of death at the midblastula stage.

Expression of c-erbB family gene products in adenoid cystic carcinoma of salivary glands: an immunohistochemical study.

The tyrosine kinase receptor family, including the epidermal growth factor receptor (EGF-R), c-erbB2 and, more recently, the c-erbB3, has been recognized as being of particular importance in many human malignancies. This study was undertaken to define the role of c-erb B2 and c-erbB3 in adenoid cystic carcinomas (A.C.C.) of the salivary glands. Sixteen cases of A.C.C. were studied immunohistochemically, using antibodies against each erbB gene family product. EGF-R was not detected in any of these samples but c-erbB2 and c-erbB3 gene products (ERBB2and ERBB3) were demonstrated in all A.C.C. sections with some degree of straining. Tubular and cribriform patterns overexpressed particularly large amounts of ERBB2 and ERBB3. Strong staining was mainly demonstrated in tumor cells of the invasive area. These results suggested that overexpression of ERBB2 and ERBB3 is related to tumor differentiation and invasion in adenoid cystic carcinomas.

Identification and disruption analysis of the recN gene in the extremely radioresistant bacterium Deinococcus radiodurans.

We isolated a radiosensitive mutant strain, KR4128, from a wild-type strain of Deinococcus radiodurans, which is known as a extremely radioresistant bacterium. The gene that restore the defect of the mutant in DNA repair was cloned, and it turned out to be the homolog of the recN gene of Escherichia coli. The recN gene encoded a protein of 58 kDa, and, in its N-terminal region, a potential ATP binding domain was conserved as expected for a prokaryotic RecN protein. An analysis of sequence of the mutant recN gene revealed a G:C to T:A transversion near the 3' end of the coding region. This alteration causes an ochre mutation, and results in the truncation of 47 amino acids from the C-terminal region of the RecN protein. The null mutant of recN gene was constructed by insertional mutagenesis, and it showed substantial sensitivities to various types of DNA damaging agents, indicating that a single defect in the recN gene can directly affect the DNA damage resistant phenotype in D. radiodurans. The recN locus of KR4128 was also disrupted and the disruptant indicated the sensitivity that was indistinguishable from its progenitor. The result indicate that the transversion in the recN gene of KR4128 cells causes a complete loss of function of the RecN protein and thus the C-terminal region of the RecN protein includes domain essential to its function.

Molecular analysis of the Deinococcus radiodurans recA locus and identification of a mutation site in a DNA repair-deficient mutant, rec30.

Deinococcus radiodurans strain rec30, which is a DNA damage repair-deficient mutant, has been estimated to be defective in the deinococcal recA gene. To identify the mutation site of strain rec30 and obtain information about the region flanking the gene, a 4.4-kb fragment carrying the wild-type recA gene was sequenced. It was revealed that the recA locus forms a polycistronic operon with the preceding cistrons (orf105a and orf105b). Predicted amino acid sequences of orf105a and orf105b showed substantial similarity to the competence-damage inducible protein (cinA gene product) from Streptococcus pneumoniae and the 2'-5' RNA ligase from Escherichia coli, respectively. By analyzing polymerase chain reaction (PCR) fragments derived from the genomic DNA of strain rec30, the mutation site in the strain was identified as a single G:C to A:T transition which causes an amino acid substitution at position 224 (Gly to Ser) of the deinococcal RecA protein. Furthermore, we succeeded in expressing both the wild-type and mutant recA genes of D. radiodurans in E. coli without any obvious toxicity or death. The gamma-ray resistance of an E. coli recA1 strain was fully restored by the expression of the wild-type recA gene of D. radiodurans that was cloned in an E. coli vector plasmid. This result is consistent with evidence that RecA proteins from many bacterial species can functionally complement E. coli recA mutants. In contrast with the wild-type gene, the mutant recA gene derived from strain rec30 did not complement E. coli recA1, suggesting that the mutant RecA protein lacks functional activity for recombinational repair.

Photorepair and excision repair removal of UV-induced pyrimidine dimers and (6-4) photoproducts in the tail fin of the medaka, Oryzias latipes.

Induction and repair of UV-B induced DNa damage in the tail fin of the Medaka, were examined immunohistochemicaly and by the enzyme-linked immunosorbent assay (ELISA). UV-induced DNA damage was detected only in the outermost layer of epithelial cells and did not differ in fishes having different degree of melanization. Both pyrimidine dimers and (6-4) photoproducts in the fin cells were removed by excision repair in the dark, the excision of (6-4) photoproducts being about twice as efficient as that of pyrimidine dimers. The rate of excision repair of UV-induced lesions in fin tissue was three to four times that in cultured Medaka cells, OL32. In the fin cells, reductions in the numbers of pyrimidine dimers and (6-4) photoproducts were seen after treatment with fluorescent light, whereas less reductions of pyrimidine dimers and no reductions of (6-4) photoproducts were observed in OL32 cells.

[Clinical effect of combined use of shouhu-san and mequitazine in recurrent vaginal candidiasis received protracted antibiotic therapy].

Clinical effects of combined use of Shouhu-san and mequitazine in 8 cases with recurrent vaginal candidiasis in obstetrics and gynecology were studied, and the following results were obtained. Shouhu-san was orally administered to 8 patients, with recurrent vaginal candidiasis, using a daily dose of 7.5 g. The duration of treatment was between 14 to 28 days. Also mequitazine was used in combination, with a daily dose of 6 mg. The clinical responses were good in all 8 and the efficacy rate was 100%. The microbiological eradication was obtained in all 8 cases of Candida albicans. Neither subjective nor objective side effects were observed. These results suggest that a combination of Shouhu-san and mequitazine appears to be useful of immunologic aspects against recurrent vaginal candidiasis.

[Clinical studies on flomoxef in the perinatal period infections].

We conducted clinical efficacy and safety tests of flomoxef (FMOX, 6315-S) in the perinatal infections and obtained the following results. 1. A total of 25 patients was treated: 16 patients with intrauterine infections, 2 patients with pelvioperitonitis, 4 patients with urinary tract infections and 3 patients with other infections. FMOX was injected at a daily dose of 2-4 g for 3-15 days (6-60 g for total dose) by intravenous drip infusion, intravenous injection or their combination. 2. The clinical efficacy rate was 96.0% of 25 patients: excellent in 4 cases (16.0%), good in 20 cases (80.0%) and poor in 1 case (4.0%). Bacteriological effects obtained were: eradicated in 14/16 cases (87.5%) replaced in 3 cases. 3. There were no subjective or objective side effects, nor were any abnormal laboratory test values attributable to the drug. From these findings, we consider that FMOX treatment appears to obtain good clinical and bacteriological responses and in safe in perinatal period infections.

[Ecological treatment of bacterial vaginosis].

Ecological treatment of bacterial vaginosis with a Lactobacillus (yoghurt) was studied, and the following results were obtained. 1. A total of 11 women aged 20 to 60 with bacterial vaginosis were treated with intravaginal application of 5 ml of commercial yoghurt (pH 4.3 +/- 0.2). The effect of the treatment was evaluated 3 days after administration by monitoring the vaginal discharge and bacteriological assessment. 2. The clinical improvement was evaluated and the decreases of vaginal discharge and vaginal redness were significant and vaginal pH was lowered significantly also (P < 0.05). In the vaginal discharge 29 strains of bacteria were detected, but 3 days after administration, all 14 strains of Gram-negative bacteria disappeared. As for the overall bacteriological effects, 6/11 cases (54.5%) were eradicated. 3 cases were partly eradicated, 2 cases were replaced. These findings indicated that the Lactobacillus therapy was effective in both clinical and bacteriological responses.

[Clinical studies on aztreonam in perinatal use].

Clinical effects of aztreonam (AZT), a monobactam antibiotic, on perinatal infections were studied with the following results: 1. Efficacy rate of AZT in the clinical application to 12 cases of perinatal infections via 2-4 g/day drip infusion (total dosage 8-24 g) was 91.7% (11/12). Breakdown of the effects according to diseases were "excellent" for mastitis (n = 3), pyelonephritis (n = 1) and urinary tract infection (n = 1), and "good" for a total of 6 cases, i.e., amnionitis (n = 2), amniotic fluid infections (n = 3) and external genital infection (n = 1). 2. Results of bacteriological studies were "eradicated" (n = 1), "replaced" (n = 4), "appeared" (n = 2) and "unknown" (n = 5). MIC values of AZT (10(6) cells/ml) were in a range of 25- greater than 100 micrograms/ml in 1 case of cervicitis (complicated with amnionitis) in the 13th week of pregnancy where AZT was found non-effective. 3. Neither subjective nor objective side effects nor abnormal clinical values were observed during the treatment with AZT.

[Influences of postoperative prophylaxis treatments on vaginal bacteria flora at hysterectomy].

We performed abdominal hysterectomy and administered aztreonam (AZT) for prophylaxis of postoperative infections, and investigated influences of the antibiotics on vaginal bacterial flora. The obtained results are summarized as follows. 1. Patients administered with AZT were studied (n = 48) for bacteria isolated from their vaginae preoperatively and at the 7th and the 14th days postoperatively. Groups of patients administered with cephalothin (CET), cefamandole (CMD) and latamoxef (LMOX) were also studied for comparison. 2. In the AZT group, the rate of isolation of aerobic Gram-positive bacteria was high (68.8-81.4%) whereas those of aerobic Gram-negative and anaerobic bacteria were low. 3. When changes in isolation frequencies of bacteria from the subjects preoperatively to postoperatively were compared, the following tendencies were observed: increase in the rate of aerobic Gram-positive bacteria was larger in the LMOX group than in the AZT group whereas decrease in the rate of aerobic Gram-negative bacteria was largest in the LMOX group followed by the AZT group then by the CMD group. Rates of decrease of anaerobic bacteria showed similar tendencies to the latter. Isolation rates of Candida sp. showed the largest increase in the LMOX group, followed by CET, CMD then AZT groups.

[Levofloxacin in obstetrics and gynecology].

We investigated the clinical efficacy and the safety of levofloxacin (LVFX, DR-3355) in obstetrical and gynecological infections and the following results were obtained. 1. Clinical efficacies were evaluated as excellent in 2 and good in 9 cases, hence the efficacy rate was 100%. 2. Bacteriologically, the eradication rate was 16/17 (94.1%). 3. No Side effects nor abnormal changes in laboratory test results were observed. These results indicated that LVFX was highly useful in the treatment of obstetrical and gynecological infections.

[Clinical effect of cefotetan against infectious diseases in obstetrics and gynaecology].

Clinical study on cefotetan (CTT), a new cephamycin antibiotic, was carried out and the following results were obtained. CTT was intravenously administered at a daily dose of 2 to 4 g to 24 cases including 10 cases with intrauterine infection, 2 cases with adnexitis, 6 cases with intrapelvic infection and 6 cases with extragenital infection. Marked response was seen in 8 cases, moderate response in 15 and no response in 1. Neither side effects nor abnormal values in clinical laboratory tests were observed.

[A study on the penetration of cefmenoxime into bile and gallbladder tissues and the pathological changes of inflamed tissues].

The penetration of cefmenoxime (CMX) into bile and gallbladder tissues was studied. A dose of 2 g CMX was given intravenously to each of patients with gallstones. Concentrations of CMX in the gallbladder tissues were 120.6 +/- 39.8 micrograms/g in normal tissues about 2 hours after the administration and 128.3 +/- 52.3 micrograms/g in inflamed tissues. The penetration of CMX into gallbladder tissue tended to decrease in proportion to the severity of inflammation, but the difference between the 2 groups, normal and inflamed tissues, were not statistically significant. The penetration of CMX into gallbladder tissue was presumed to increase in proportion to increase in the serum concentration of CMX. The result indicated that the achievement of high concentrations of CMX in the serum is important in the treatment of biliary tract diseases.

[A clinical study on the efficacy of ceftazidime alone or in combination with aspoxicillin against infections in obstetric and gynecological patients].

We studied clinical effects of ceftazidime (CAZ) alone or in combination with aspoxicillin (ASPC) against various infections in obstetric and gynecological patients. 1. Obstetric and gynecological patients (n = 91) with various infectious diseases were treated with CAZ alone (1-2 g x 2/day, n = 54) or in combination with ASPC (1-2 g x 2/day, n = 37) administered via drip infusion. 2. CAZ alone or in combination with ASPC was efficacious in 50 out of 54 (92.6%) or 33 out of 36 (91.7%) patients, respectively. Overall, the efficacy ratios were 46/49 (93.9%) against gynecological infections, 21/25 (84.0%) against perinatal infections and 16/16 (100%) against other infections. The bacteriological efficacy ratio was 21/21 (100%) while clinical effectiveness in cases in which causative agents were known was observed in 20 out of 21 (95.2%) patients. In patients who had not respond to other treatments, CAZ alone, and in combination with ASPC were effective in 15 out of 16 (93.8%) and 6 out of 8 (75.0%) patients, respectively, hence the overall efficacy ratio was 21/24 (87.5%). 3. Abnormal values in clinical laboratory tests were obtained in 3 out of 91 (3.3%) patients. No other adverse side effects were observed in any of the patients.

[Clinical effect of the combined use of aztreonam, amikacin and clindamycin in infectious disease in obstetrics and gynecology].

Clinical effects of combined use of aztreonam (AZT), amikacin (AMK) and clindamycin (CLDM) in 46 cases with infectious diseases in obstetrics and gynecology were retrospectively studied in 2 groups, and the following results were obtained. 1. No significant difference in efficacy rates was noted between AZT plus CLDM treated group (n = 25) and AMK plus CLDM treated group (n = 21) (96.0% vs. 95.2%), while rate of excellent efficacy was slightly higher in AZT plus CLDM group than AMK plus CLDM group (24.0% vs. 14.3%). 2. No significant difference in bacteriological clinical effects was also noted between the 2 groups, while bacteriological eradication rate was higher in the AZT plus CLDM group than in the other group (76.2% vs. 50.0%), and the difference was particularly clear in the eradication rates of aerobic, Gram-negative bacteria (88.9% vs. 30.0%). 3. Subjective and objective side effects, and abnormalities of clinical test results were not found in either group.

Comparisons of the bacterial flora in genital regions at non-pregnancy.

In the development of infectious diseases at non-pregnancy and at pregnancy, correlations between bacterial flora in the vagina and portio vaginalis and the ascending infections of those bacteria have recently been discussed. To clarify the cause of those infectious diseases, we studied the localization of microorganisms in genital regions. Patients undergone abdominal total hysterectomy (n = 172) were employed as subjects, and microorganisms isolated from 4 genital regions were studied. In addition, the preventive effect of cefmetazole (CMZ) against postoperative infections was analyzed in 479 cases including the hysterectomy cases mentioned above. The results obtained are summarized as follows: 1. The isolation rate of microorganisms at non-pregnancy, from subjects of 30 to 69 years old, was 65.6% (82/125) in the vagina and portio vaginalis, 52.1% (25/48) in the cervical mucus, 7.3% (9/124) in the uterine cavity and 0% (0/47) in the ovarian surface. 2. Numbers of microorganisms isolated in each region were 99 strains in the vagina and portio vaginalis, 28 in the cervical mucus, 10 in the uterine cavity and none in the ovarian surface. Isolation of aerobic Gram-positive bacteria (60-89.3%) and anaerobic Gram-positive bacteria (7.1-30%) were varied in each region. Lactobacillus spp. (38 strains), Staphylococcus epidermidis (20 strains) and Propionibacterium acnes (10 strains) were isolated from vagina and portio vaginalis, and Lactobacillus spp. (17 strains) were the most often isolated bacteria from the cervical mucus.(ABSTRACT TRUNCATED AT 250 WORDS)