Developmental enhancement of alpha2-adrenoceptor-mediated suppression of inhibitory synaptic transmission onto mouse cerebellar Purkinje cells.

Noradrenaline (NA) modulates glutamatergic and GABAergic transmission in various areas of the brain. It is reported that some alpha2-adrenoceptor subtypes are expressed in the cerebellar cortex and alpha2-adrenoceptors may play a role in motor coordination. Our previous study demonstrated that the selective alpha2-adrenoceptor agonist clonidine partially depresses spontaneous inhibitory postsynaptic currents (sIPSCs) in mouse cerebellar Purkinje cells (PCs). Here we found that the inhibitory effect of clonidine on sIPSCs was enhanced during postnatal development. The activation of alpha2-adrenoceptors by clonidine did not affect sIPSCs in PCs at postnatal days (P) 8-10, when PCs showed a few sIPSCs and interneurons in the molecular layer (MLIs) did not cause action potential (AP). In the second postnatal week, the frequency of sIPSCs increased temporarily and reached a plateau at P14. By contrast, MLIs began to fire at P11 with the firing rate gradually increasing thereafter and reaching a plateau at P21. In parallel with this rise in the rate of firing, the magnitude of the clonidine-mediated inhibition of sIPSCs increased during postnatal development. Furthermore, the magnitude of the clonidine-mediated firing suppression in MLIs, which seemed to be mediated by a reduction in amplitude of the hyperpolarization-activated nonselective cation current, I(h), was constant across development. Both alpha2A- and alpha2B-, but not alpha2C-, adrenoceptors were strongly expressed in MLIs at P13, and P31. Therefore, the developmental enhancement of the clonidine-mediated inhibition of sIPSCs is attributed to an age-dependent increase in AP-derived sIPSCs, which can be blocked by clonidine. Thus, presynaptic activation of alpha2-adrenoceptors inhibits cerebellar inhibitory synaptic transmission after the second postnatal week, leading to a restriction of NA signaling, which is mainly mediated by alpha1- and beta2-adrenoceptors in the adult cerebellar neuronal circuit.

Cell death with predominant apoptotic features in Saccharomyces cerevisiae mediated by deletion of the histone chaperone ASF1/CIA1.

BACKGROUND: Although no potential homologues of multicellular apoptotic genes (e.g. Bax, Bak, Bcl-2, caspases and p53) have been identified in a unicellular eukaryote, previous reports contain several implications of the apoptotic behaviour of yeasts (i.e. Saccharomyces cerevisiae and Schizosaccharomyces pombe). Therefore, whether or not yeast undergoes apoptosis has been a topic of some debate. hCCG1, which is the largest subunit of TFIID and a histone acetyltransferase, appears to be involved in the regulation of apoptosis. The factor hCIA interacts with hCCG1 and functions as a histone chaperone in mammalian cells; its homologue in yeast is Asf1p/Cia1p. Therefore, we anticipated that a yeast mutant in Asf1p/Cia1p would be a valuable tool for studying apoptosis in yeast. RESULTS: We established a strain of S. cerevisiae lacking the histone chaperone ASF1/CIA1. This disruptant, asf1/cia1, arrested preferentially at the G2/M-phase and died. We systematically analysed the phenotype associated with the death of this mutant yeast and identified many changes, such as fragmentation of the nuclei, condensation and fragmentation of chromatin, reduction of the mitochondrial membrane-potential, dysfunction of the mitochondrial proton pump, and a discernible release of cytochrome c to cytoplasm that resembles those in apoptotic multicellular organisms. Other changes potentially associated with the death in our mutant included a reduction in the vacuolar membrane potential, dysfunction of the vacuolar proton pump, reduction of endocytosis, and the presence of many autophagic bodies. However, these mutant yeast cells also showed cellular enlargement, which is characteristic of necrosis. CONCLUSIONS: Cell death in S. cerevisiae occurs with a phenotype that largely resembles apoptosis in multicellular organisms, but that has some features of necrosis. Therefore, we indicate that yeast undergoes a 'prototypal active cell death' that retains some characteristics of passive cell death (necrosis). In addition, we think that active cell death is ubiquitously the essential attribute of life. Although such an active cell death system in yeast remains open to confirmation, we speculate that deletion of the histone chaperone Asf1p/Cia1p inhibits the normal assembly/disassembly of nucleosomes in yeast and thereby initiates the active cell death system.

[Pyogenic sacroiliitis in pregnancy].

Pyogenic sacroiliitis is an extremely rare disease in the field of obstetrics and gynecology, and especially for the cases discovered and treated during pregnancy, only five cases can be found in literature. We experienced one case of the disease in which the patient needed urgent hospitalization due to dysbasia caused by fever and pain at the left hip at the 27th week of her pregnancy. The patient was a 31-year-old primipara presenting typical clinical symptoms of pyogenic sacroiliitis along with evidence of severe infection as represented by fever of 39.7 degrees C and CRP of 12.6 mg/dl. She showed a good response to meropenem (MEPM) at 1 g twice a day for 8 days and then at 0.5 g twice a day for 2 days, followed by faropenem (FRPM) at 200 mg three times a day for 12 days, which successfully improved her subjective and objective findings as well as her laboratory test values, resulting in a complete cure. The definitive diagnosis of the disease in the patient was made on the basis of MRI findings, but no pathogen was identified. The patient was found to have marginal placenta previa as a complication, but she had an uneventful trans-vaginal delivery at the 37th week of her pregnancy and left hospital after both she and her baby showed favorable post-delivery progress. The case reported here is the first case of pyogenic sacroiliitis that has ever been discovered and treated during pregnancy in Japan.

[Clinical investigation of cefpirome for perinatal infection and infection prevention].

Effectiveness and safety of cefpirome (CPR) were studied in perinatal infection and prevention of postoperative infections. 1. Enrolled in the study of perinatal infections were the total 62 cases comprising 47 of chorioamnionitis, 10 of puerperal intrauterine infections, 1 of infectious abortion and 4 of others. The effective rate of CPR 2 g/day given in drip infusion was 61/62 (98.4%). In bacteriological studies, the bacterial elimination rate was 57/61 (93.4%). Unchanged and remained were 3 strains of Gram-positive bacteria and 1 strain of Bacteroides spp. 2. Enrolled in the study of prevention of postoperative infections during perinatal period were the total 88 cases comprising 74 who underwent cesarean section and 14 others. The effective rate of CPR 2 g/day given in drip infusion was 87/88 (98.9%). 3. With respect to subjective and objective adverse affects due to the agent, drug eruption was observed in 1/150 (0.7%). No abnormal result of clinical laboratory tests was found. The above results suggested the usefulness of CPR for treatment of perinatal infections and prevention of postoperative infections.

[Clinical effects of meropenem on infectious diseases in obstetrics and gynecology].

The efficacy and the safety of meropenem (MEPM) were examined in obstetric and gynecological infections and the results shown below were obtained. 1. The subjects were patients with infectious diseases during a non-pregnant period (n = 14), a pregnant period (n = 13) and a puerperal period (n = 11). After intravenous drip infusion of MEPM at 1-2 g/day, the response rate was 38/39 (97.4%) (Excellent: 13/39 (33.3%), Effective: 25/39 (64.1%)). 2. The response rates in a group receiving 1 g/day, a group receiving 2 g/day and a group receiving a combination of both were, respectively, 25/26 (96.2%), 9/9 (100%) and 4/4 (100%). 3. In cases that did not respond to previous treatments, the response rate to MEPM was 20/20 (100%). 4. In clinical efficacy classified by causative organisms, the rate of Excellent was 10/23 (43.5%) and the rate of Effective was 13/23 (56.5%), and the bacterial eradication rate was 21/23 (91.3%). 5. No subjective and objective adverse reactions or abnormalities in clinical laboratory tests due to administration of MEPM were observed.

[Clinical effects of cefoselis (CFSL) on infections in obstetric and gynecologic field and prevention of postoperative infections].

Clinical effects of cefoselis (CFSL) on various infections and prevention of postoperative infections in the field of obstetrics and gynecology were investigated with a total of 100 patients of 8 facilities in Yamagata and the following results were obtained: 1. For the patients (n = 70) who received the treatment with CFSL at 2 g/day for 5 days to prevent postoperative infections, the treatment was effective for such infections in 68 of 69 (98.6%) except one to whom the treatment was discontinued during the course. 2. For the patients with infections (n = 30) who were treated with CFSL at 2-4 g/day for 5-7 days, the treatment was markedly effective in 8/30 (26.7%), effective in 21/30 (70%) and not effective in 1/30 (3.3%). The overall rate of efficacy was 29/30 (96.7%). Based on the clinical effects for each isolate, the bacteriological efficacy was evaluated as 29/29 and the rate of bacterial eradication for each isolate was 23/29 (79.3%). 3. Laboratory test revealed liver functional abnormalities in one patient and eruption, a subjective/objective symptom caused by CFSL was noted in two patients. These results suggest that CFSL is effective for various infections in obstetric and gynecologic field and also the prevention of postoperative infections.

[Clinical studies of faropenem in the field of obstetrics and gynecology].

The clinical effect of faropenem was evaluated in 165 ambulatory patients with various infections in the field of obstetrics and gynecology at 10 institutions in Yamagata Prefecture. The results obtained are summarized below. 1. The rate of efficacy, as determined from the clinical effect following 3- to 7-day repeated administration at a dose of 600 mg/day, was 97.9% (46/47) for intrauterine infections, 92.0% (23/25) for adnexitis, 93.8% (15/16) for external genital infections, 88.9% (8/9) for mastitis, 94.0% (63/67) for cystitis, and 100% (1/1) for cervicitis. The overall efficacy rate was estimated to be 94.5% (156/165). 2. The rate of clinical efficacy, as classified by isolate, was high, 95.1% for Gram-positive bacteria, 100% for Gram-negative bacteria, and 100% for anaerobes. As for bacteriological response classified by isolate, the eradication rate was high, 91.4% (74/81) for Gram-positive bacteria, 98.4% (62/63) for Gram-negative bacteria, 89.5% (17/19) for anaerobes, and 93.9% (153/163) in all. 3. No adverse reactions or laboratory abnormalities were observed in any patient. The results presented suggest that faropenem is a highly safe and effective antibiotic for the treatment of obstetric or gynecological infections of various kinds in an ambulatory setting.

[Change of cytokines and clinical efficacy of panipenem/betamipron in obstetric and gynecological infections. Yamagata Study Group of Panipenem/Betamipron in Obstetric and Gynecological Infections].

Efficacy and safety of panipenem/betamipron (PAPM/BP) in treatment of obstetric and gynecological infections, and change of interleukin-6 (IL-6) and interleukin-8 (IL-8) levels in blood, as markers of infection, were investigated. The results were as follows; 1) Clinical efficacy of PAPM/BP by drip infusion of 1-2 g/day for 3-14 days against 52 patients with intrauterine infection (n = 29), pelveoperitonitis (n = 19), and other infections were 14 "Excellent" in 14 cases, "Good" in 35 cases, and efficacy rate was 94.2% (49/52). Both efficacy rate analy by causative organisms and eradication rate were 35/37 (94.6%). No subjective or objective side effects and no abnormal labolatory findings were observed. 2) Changes of IL-6 (> 4 pg/ml) levels in serum, as an infection marker, were observed in 8 cases out of 14 cases (57.1%), and correlation between CRP and IL-6 in the treatment process was noticed. However, changes of serum IL-8 (> 12.5 pg/ml) were observed in only 2 cases of those 14 cases (14.3%), indicative that IL-8 has no significance as a marker of infection.

[Ecological treatment of bacterial vaginosis and vaginitis with Bio-three].

Ecological treatment of bacterial vaginosis and vaginitis with a Bio-three was studied, and the following results were obtained. 1. A total of 16 women with bacterial vaginosis and vaginitis were treated with intravaginal application of 2 g of Bio-three (E. faecalis T-110, C. butyricum TO-A, B. mesentericus TO-A, pH 6.9 +/- 0.3). 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 (5.29 +/- 0.24 vs. 4.31 +/- 0.37, p < 0.05). In the vaginal discharge 35 strains of bacteria were detected, but 3 days after administration, 16/30 strains of Gram-positive bacteria, and 2 strains of Gram-negative bacteria disappeared. As for the overall bacteriological effects, 7/16 cases were eradicated, 1 case was partly eradicated, 6 cases were replaced. These findings indicated that the Bio-three therapy was effective in both clinical and bacteriological responses.

[Intravaginal bacterial flora and clinical significance of granulocyte elastase and pH determination].

In 95 patients with abnormal vaginal and cervical secreta (49 pregnant women and 46 non-pregnant women), the relation between intravaginal flora and intravaginal granulocyte elastase (Elastase) and pH was investigated. The results were as follows. 1) Gram-positive bacteria were detected in the vaginal secreta at a high rate (87/144, 60.4%), and it mainly consisted of Lactobacillus sp. (67/142, 46.5%). It was followed anaerobia (26/144, 18.1%) and fungi (26/144, 18.1%). 2) The patients with cervicitis or vaginitis had higher elastase value (6.65-6.69 micrograms/ml) than the ones with vaginal erosion, and the patients who showed an intravaginal pH value not lower than 5.0 had significantly increased elastase value (6.44 +/- 1.40 micrograms/ml) than the patients who showed the values 4.5 or higher. 3) Regarding the relation between the detected bacteria and elastase values, elastase values were higher in the patients infected by anaerobia (6.58 +/- 1.40 micrograms/ml), Gram-negative bacteria (6.01 +/- 3.61 micrograms/ml), Gram-positive bacteria (5.02 +/- 0.94 micrograms/ml) and fungi (5.14 +/- 1.08 micrograms/ml) than the values in patients with Lactobacillus sp. (pH < 4.5). Further, the intravaginal pH value was higher than 4.5 in all of these groups, which was higher compared with the one in the patients infected with Lactobacillus sp. (4.04 +/- 0.04).

[Measurement of antibody titers to chlamydial infection and effects of levofloxacin in cystic cervicitis and chlamydial infection].

Generally, clinical symptoms such as abnormal leukorrhea are caused by C. trachomatis, an ordinary bacteria in cervical infection. The effects of levofloxacin administration at a dose of 300 mg/day for 5-14 days were investigated in the subjects (n = 66) after the discussion of cystic cervicitis. The treatment was made in combination with chloramphenicol-solcoseryl tablet for vaginal use. And it was demonstrated that treatment was effective in all subjects. Then, Sero IPALISA, an examination of IgA/IgG antibody was conducted for the screening of Chlamydia infection (n = 258). The rate of antibody-positive case was 48/160 (30.0%) for the non-pregnant women and 26/98 (26.5%) for the pregnant. The occurrence rates for the women singing in 15-24 and 35-39 years of age were 50 and 41%, respectively. The results from the measurement of the antibody titer were as follows: the rate of IgA/IgG positive care was 61/87 for IgA and 56/87 for IgG when the cut-off index was 1.11 or more. The rates for both antibody were 11/87 (12.6%) and 24/87 (27.6%) for the indexes of 1.11-3.00 and 3.01 or more, respectively. Next, one to three courses levofloxacin at 300 mg/day for 14 days were given to 48 non-pregnant subjects infected with Chlamydia and one to two courses of clarithromycin at 400 mg/day for 14 days were given to 26 pregnant subjects. Side effects have not been noted in any care and there was no changes in the IgA/IgG antibody titer depending on these treatments.

Concentration of levofloxacin in cervical mucus and its clinical effects on cervicitis.

An investigation was made on the concentration of levofloxacin (LVFX) in cervical mucus and its clinical effects on cervicitis. The results were as follows: 1) The concentrations of orally administered LVFX in the cervical mucus of 110 subjects were determined by HPLC. During 1-4 hour after the administration the mean concentration of LVFX in the cervical mucus reached a level of 2 micrograms/g, which was higher than the serum level. The transfer of LVFX to the cervical mucus was almost the same as that to other genital organs. 2) When LVFX was given to 102 patients at a dose of 100-200 mg, t.i.d for 4-5 days and the efficacy was evaluated with clinical improvement, the clinical efficacy rate of LVFX was 72/102 (70.6%). Significant bacteriological effects were observed in 70/73 (95.9%), especially, the disappearance rate of C. trachomatis was 18/18 (100%). 3) The administration LVFX did not cause any subjective or objective side effects and any abnormalities were not detected in the laboratory test done in this study. These results demonstrate that LVFX can be sufficiently transferred to the cervical mucus for the treatment of cervicitis due to the infection of C. trachomatis etc.

[Clinical study of tazobactam/piperacillin in obstetrics and gynecological infections].

A clinical study was carried out to investigate clinical efficacy and safety of tazobactam/piperacillin (TAZ/PIPC), a newly developed antibiotic agent for injections. The obtained results are as follows: 1. In 10 patients with obstetrics and gynecological infections (7 patients with endometrial infections, one patient with pelvic peritonitis, one patient with ovarian abscess and one patient with Bartholins's abscess) intravenous drip infusion of TAZ/PIPC, 5.0 g/day, was administered for 5 to 10 days. The total doses were 2550 g/body. 2. Clinical efficacy was evaluated in the above cases. As a result, an excellent result was obtained in one patient and good efficacies in 9 patients. The efficacy rate was 100% (10/10). Concerning the clinical bacteriological effect, eradication ratio was 83.3% (5/6). Concerning the bacteriological effect against 11 strains of clinical isolates, 4 strains were eradicated, 6 strains were replaced and one strain was unchanged. 3. Neither subjective/objective side effects nor abnormal laboratory test results were observed upon administration of TAZ/PIPC. These results suggested the usefulness of tazobactam/piperacillin in obstetrics and gynecological infections.

Placental transfer of lidocaine hydrochloride after prolonged continuous maternal intravenous administration.

We treated a patient with arrhythmia during pregnancy with prolonged intravenous administration of lidocaine hydrochloride. This was a case of twin-to-twin transfusion syndrome and the arrhythmia was caused by ritodrine therapy. In total, 14.1 g lidocaine (50 mg.hr-1 for 282 hr) were used. Since there are no descriptions of human placental transfer of lidocaine after such a prolonged continuous intravenous administration, we measured lidocaine concentrations in maternal and fetal serum, and in the amniotic fluid (AF) at delivery. Fetal serum lidocaine concentrations (donor: 0.83 microgram.ml-1; recipient: 0.82 microgram.ml-1) were lower than in the maternal serum (1.6 micrograms.ml-1), while the AF lidocaine concentrations (donor: 1.05 micrograms.ml-1; recipient: 1.04 micrograms.ml-1) were higher than those of the fetal sera. The fetal/maternal concentration ratios of lidocaine were 0.52 for the donor and 0.51 for the recipient, which were similar to those described previously after administration of lidocaine in labour.

[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.

[A clinical study on chemotherapies for chorioamnionitis].

Effects of imipenem/cilastatin (IPM/CS) therapy, flomoxef (FMOX) therapy and combined ceftazidime + aspoxicillin (CAZ/ASPC) therapy as initial therapies for chorioamnionitis were assessed clinically. 1. The subjects were 49 women with threatened abortion and 29 with premature rupture of membranes (PROM), complicated in all cases by chorioamnionitis. The inflammation was treated with IPM/CS in 19 patients, FMOX in 39, CAZ in 11, and CAZ/ASPC in 9. 2. The response rate to therapy for chorioamnionitis was 95.9% (47/49) in the threatened abortion group. Of the 49 patients in this group, 16 (32.7%) underwent premature labor. Of the therapies administered, IPM/CS tended to prevent premature labor more frequently than did any other therapy. The latent period (from rupture of membranes to delivery) was equal to or longer than 7 days in the PROM group. The percent prolongation of the latent period in these patients (55.5%) was significantly greater than that previously obtained with penicillin therapy. 3. The bacterial elimination rate was 50.9% (29/57). Of the 36 bacterial isolates, 66.7% were Gram-positive bacteria. The bacteriological efficacy rate was 89.7% (26/29). These results suggest that antibacterial agents effective against Gram-positive bacteria should be selected for treatment of chorioamnionitis, and that IPM/CS therapy is particularly useful considering the drug's good transfer into amniotic fluid and its antibacterial spectrum.

[A clinical study on the efficacy of ceftazidime and aspoxicillin in chorioamnionitis. Abdominal Infections Research Group].

Chorioamnionitis as a complication of threatened abortion and preterm labor and preterm PROM were treated with ceftazidime (CAZ) and aspoxicillin (ASPC) as a primary therapy. The following results were obtained. 1. Cases of threatened abortion and preterm labor (n = 25) and preterm PROM (n = 5) were treated with 2-4 g CAZ/day alone (n = 14) or in combination with 4 g ASPC/day (n = 16) along with a uterine contraction inhibitor (ritodrine hydrochloride etc. n = 28) and clinical evaluation was made. 2. In the cases of threatened abortion and preterm labor the efficacy ratio was 24/25 (96%). In the cases of preterm PROM, the latent period-delaying effect was observed in five out of the five patients. Upon analysis of the tocolysis index in the group of threatened abortion and preterm labor, the index values > or = 5 were observed in 12 out of 25 (60%), and the delivery incidence before the 35th week of gestation was 4/25 (16%). In all patients, the incidence of delivery after the 36th week of gestation was 24/30 (80%). 3. Bacteriological examinations showed a high detection rate for Gram-positive bacteria, and the combination effect between ASPC and CAZ was demonstrated against all 9 isolates examined. The above results indicated a high efficacy as well as safety of the combination of CAZ and ASPC as a primary therapeutic means against chorioamnionitis.

[Evaluation of changes in IL-6 in gynecological infections].

We administered imipenem/cilastatin sodium (IPM/CS) to patients with gynecological infections as initial treatment and evaluated changes in blood interleukin 6 (IL-6) as an infection marker. 1. The subjects consisted of 7 patients with chorioamnionitis, 4 with intrauterine infections, and 1 with subcutaneous abscess. IPM/CS (1-2 g/day) was intravenously drip infused. This therapy was markedly effective in 1 patient and effective in 11; the response rate was 100%. 2. IL-6 generally began to decrease earlier than CRP. Before drug administration, correlations were observed between IL-6 and CRP (r = 0.946) between IL-6 and elastase (r = 0.355), and between elastase and CRP (r = 0.579). During the entire course, correlations were observed between IL-6 and CRP (r = 0.581), between IL-6 and elastase (r = 0.303), and between elastase and CRP (r = 0.776). These results suggest that blood IL-6 reflects early the pathologic state and treatment effects, and is a useful infection marker in gynecological infections.

[Clinical effects of imipenem/cilastatin sodium on gynecological infections].

We evaluated the effectiveness and safety of imipenem/cilastatin sodium (IPM/CS) in gynecological infections at gynecological departments at multiple institutions in Yamagata Prefecture. The subjects consisted of 64 patients with gynecological infections, 9 with urinary tract infections, and 21 with other infections. IPM/CS was intravenously drip infused at a dose of 1-2 g/day. The overall response rate was 97.9% (92/94); 96.9% (62/64) for gynecological infections, 100% (9/9) for urinary tract infections, and 100% (21/21) for other infections. Bacteriologically the response rate was 97.3% (36/37), and the eradication rate was 97% (32/33). As for subjective and objective side effects, only diarrhea was observed in 1 patient. Clinical laboratory examinations showed no abnormal values. These results suggest the usefulness of IPM/CS for gynecological infections.