Homogenous simultaneous detection of HCV RNA and internal control by two-color fluorescence real-time monitoring of isothermal sequence amplification with INAF probes.

We demonstrated the homogenous simultaneous detection of HCV RNA and internal control by two-color fluorescence real-time monitoring of isothermal sequence amplification with INAF probes.

Homogeneous detection of a target nucleic acid sequence by combination of the intercalation activating fluorescence DNA probe and the isothermal sequence amplification.

We demonstrated the homogeneous real-time detection of RNA produced during isothermal TRC amplification by INAF probe.

Studies on the epitope of pig zona pellucida recognized by a fertilization-blocking monoclonal antibody.

Heat solubilized pig zona pellucida (pZP) was separated into four major glycoprotein families (pZP1, pZP2, pZP3, pZP4). Cloning of a full-length cDNA coding for pZP1 revealed that the coding sequence for pZP4 was immediately followed by the sequence for the amino-terminal residue of pZP2 and that they shared a significant similarity with mouse and human ZP2, which are considered to serve as a secondary sperm receptor in sperm-zona interactions. One of the monoclonal antibodies, mAb-5H4, produced against pZP4 inhibited the binding of boar and human spermatozoa to each of the homologous oocytes. The epitope recognized by mAb-5H4 was determined to be present on a 10 amino acid sequence of pZP1(50-59) by using epitope mapping and analysis of chain flexibility of pZP4. A synthetic 18mer peptide corresponding to pZP1(50-67) reacted with mAb-5H4, and mouse antisera raised to the synthetic 18mer peptide recognized intact pig zona pellucida and strongly inhibited the fertilization of pig oocytes with boar spermatozoa in vitro. These results suggest that the pZP4 peptide (50-59) identified as an epitope for mAb-5H4 could be a promising candidate in the development of a contraceptive vaccine.

Cloning of a cDNA coding for porcine zona pellucida glycoprotein ZP1 and its genomic organization.

The zona pellucida composed of three or four glycoproteins plays important roles in fertilization. Our previous study showed that porcine ZP1, one of the major glycoproteins of porcine zona pellucida, was divided into two components (porcine ZP4 and ZP2), and suggested it was a homologue of mouse ZP2. In this paper we report the cloning of a cDNA for porcine ZP1 and its genomic organization. The deduced amino acid sequence of porcine ZP1 shared a 54% and 63% identity with those of mouse and human ZP2, respectively. Genomic organization of porcine ZP1 was also similar to that of mouse ZP2. The transcript of porcine ZP1 gene was detected only in growing oocytes.

Extractive-spectrophotometric determination of trace iron(II) with di-2-pyridylmethanone 2-(5-nitro)pyridylhydrazone.

The optimum conditions for the extractive-spectrophotometric determination of trace iron(II) with di-2-pyridylmethanone 2-(5-nitro)pyridylhydrazone have been established. Iron(II) reacts with this reagent at pH 2.0-7.5 to form an uncharged 1:2 (metal-to-ligand) complex, which can be extracted with toluene. Beer's law is obeyed over the range up to 0.84 mug/ml of iron(II) at 505 nm. The molar absorptivity of the extracted species is 5.83 x 10(4) 1.mole(-1).cm(-1). The proposed method is extremely sensitive and reproducible, and has been satisfactorily applied to the determination of total iron in freshwater samples by adding ascorbic acid to reduce iron(III).

[Hyperthermic perfusion therapy using peplomycin for bladder cancer].

Hyperthermic intravesical perfusion therapy using peplomycin (40 micrograms/ml) in distilled water at 43 degrees C as a perfusate was performed for 2 to 3 hours in 18 patients with superficial bladder tumors and 2 with deep bladder tumors. The therapeutic efficacy was determined by cystoscopy, ultrasonography and/or CT scan. Complete and partial tumor regression was obtained in 1 and 3 of the 18 patients, respectively. There was no tumor regression in the 12 patients. Most of the patients studied had bladder discomfort such as irritation, urinary frequency and so on, during and/or after perfusion. None of the patients developed acute pyelonephritis.

Multiple regression analysis of the cholinesterase activity with certain physiochemical factors.

Nineteen hundred sixteen agricultural workers who have been living in a rural area in central Japan were studied. Their age, blood pressure, Broca index, hemoglobin content, serum total cholesterol, and activity of serum cholinesterase and transaminase (GOT and GPT) were determined. The relation between the cholinesterase activity and certain physiochemical factors of the subjects was studied. Cholinesterase activity (ChE) is related to certain factors such as age, hemoglobin content (Hb), serum total cholesterol (TCh), transaminase activity (GPT), and Broca index (BI). A multiple regression equation exists between the cholinesterase activity and these factors: ln ChE = a (age) + b (Hb2) + c X ln GPT + d X ln TCh + e BI + f, where a, b, c, d, e, and f are constants. The estimated value of cholinesterase activity agrees with its measured activity. The partial correlation coefficients of the equation can be classified into the following three classes: (1) The partial correlation coefficients of total cholesterol and Broca index are nearly constant without distinction of sex and season. (2) The coefficient of hemoglobin content has a small seasonal and sexual difference. (3) The coefficients of age and GPT have a great seasonal and sexual difference. Using the equation, the most probable value of cholinesterase activity can be estimated. Therefore, the significant changes of its activity may be attributed to the toxic effects of insecticides or the abnormality of liver function.

[A double blind clinical trial of cefamandole and cefmetazole in complicated urinary tract infections].

We conducted a randomized double blind comparison of cefamandole (CMD) and cefmetazole (CMZ) in the treatment of 193 patients with complicated urinary tract infections. The patients received 1 gram of CMD or CMZ twice a day intravenously by drip infusion over 1 hour for 5 days. Pretreatment urinary leukocyte counts and urinary bacterial counts were at least 5 cells/hpf and 10(4) bacteria/ml, respectively. Each patient was randomly allocated either to CMD or CMZ group. There were 93 patients in CMD group and 100 patients in CMZ group. Clinical efficacy was evaluated based on the effect of treatment on bacteriuria and pyuria according to the criteria set by the UTI Committee, Japan. The response to CMD treatment was excellent in 18 cases (19.4%), moderate in 38 cases (40.9%) and poor in 37 cases (39.8%) with an overall effectiveness of 60.2%, whereas the response to CMZ was excellent in 19 cases (19.0%), moderate in 40 cases (40.0%) and poor in 41 cases (41.0%) with an overall effectiveness of 59.0%. No statistical significant difference was found between 2 treatment groups. Comparison of the bacteriological response between 2 groups showed that the eradication rate for strains of Gram-positive cocci were significantly higher in those patients treated with CMD. Gram-negative rods were eradicated from 68.4% of cases treated with CMD, and 78.0% of those with CMZ, but the difference was not significant. Adverse reactions were observed in 3 patients receiving CMD 1 case each of diarrhea, eruption and epigastric pain. Abnormality in laboratory tests was found in 6 patients in each treatment group. The results indicate that CMD is effective, safe and useful in the treatment of patients with complicated urinary tract infections, and its efficacy, safety and usefulness are comparable with those of CMZ.

Inhibitory effects of various drugs on dual asthmatic responses in wheat flour-sensitive subjects.

In order to investigate the mechanism of late asthmatic response (LAR), inhibitory effects of various drugs for LAR were examined in two wheat flour-sensitive asthmatic subjects who showed immediate and late responses in the allergen provocation test and skin test. Antihistamines did not inhibit the LAR but totally or partially inhibited the immediate response. By contrast, corticosteroids inhibited the LAR but not the immediate response. Disodium cromoglycate inhibited both responses. Diethyl carbamazine citrate, an inhibitor of release of SRS-A, seemed to shorten the duration of the LAR, although it has no effect on the immediate response and/or on the severity of the LAR. Indomethacin and acetyl salicylate, which inhibit prostaglandin synthesis, had no significant effects on either the immediate or the LAR.