Differential diagnosis of schistosomiasis mekongi and trichinellosis in human.

An indirect (plate) ELISA and, a more convenient version, a dot-blot (membrane) ELISA have been developed using haemocyanin of a mollusk, Megathura crenulata, i.e. keyhole limpet haemocyanin (KLH) and purified, specific antigen of Trichinella spiralis (APTsAg) obtained from a monoclonal antibody-affinity column chromatography, for differential diagnosis of schistosomiasis mekongi and trichinellosis. Serum samples of patients with parasitologically confirmed trichinellosis were reactive to both antigens in both versions of ELISA while sera of patients with schistosomiasis mekongi were positive only to the KLH. Both ELISA were negative when used to test sera of normal controls and patients with gnathostomiasis, paragonimiasis and opisthorchiasis.     

Prevalence and Characteristics of eae-Positive Escherichia coli from Healthy Cattle in Japan

The prevalence of eae-positive Escherichia coli (eaeEC) in Japan was examined using rectal stool samples taken from 35 calves less than 1 month old, 107 calves more than 1 to 3 months old, 88 heifers more than 3 to 6 months old, 214 heifers over 6 months old, and cows from 95 farms. Screening with eae PCR revealed the prevalence to be, with increasing age, 31.4, 8.4, 26.1, and 14.5%, respectively. Of 51 selected eaeEC strains, more than 40% were serotyped as O26, O103, O111, O145, or O157, which are frequently detected as enterohemorrhagic E. coli types. Four strains were identified as recently reported intimin types η, ι, and κ.     

Engineering of Polyploid Saccharomyces cerevisiae for Secretion of Large Amounts of Fungal Glucoamylase

We engineered Saccharomyces cerevisiae cells that produce large amounts of fungal glucoamylase (GAI) from Aspergillus awamori var. kawachi. To do this, we used the δ-sequence-mediated integration vector system and the heat-induced endomitotic diploidization method. δ-Sequence-mediated integration is known to occur mainly in a particular chromosome, and the copy number of the integration is variable. In order to construct transformants carrying the GAI gene on several chromosomes, haploid cells carrying the GAI gene on different chromosomes were crossed with each other. The cells were then allowed to form spores, which was followed by dissection. Haploid cells containing GAI genes on multiple chromosomes were obtained in this way. One such haploid cell contained the GAI gene on five chromosomes and exhibited the highest GAI activity (5.93 U/ml), which was about sixfold higher than the activity of a cell containing one gene on a single chromosome. Furthermore, we performed heat-induced endomitotic diploidization for haploid transformants to obtain polyploid mater cells carrying multiple GAI genes. The copy number of the GAI gene increased in proportion to the ploidy level, and larger amounts of GAI were secreted.     

Serum levels of macrophage colony-stimulating factor in singleton and twin pregnancy.

Macrophage colony-stimulating factor (M-CSF) is present in the placenta and has been implicated in placental growth and development. M-CSF levels in peripheral blood increase significantly with progression of pregnancy in uncomplicated singleton pregnancy. This study investigated whether serum M-CSF levels in singleton pregnancy differed from those in twin pregnancy. A hundred and four pregnant women, of whom fifty-nine were women carrying single fetuses and forty-five were women with twin fetuses, participated in the study. Their average gestational age at entry was 10, 20, 30, and 38 weeks of gestation in singleton pregnancy and 10, 20, 30, and 35 weeks in twin pregnancy. Peripheral blood was collected and serum was separated after centrifugation and stored at - 20 degrees C. M-CSF levels were determined by ELISA. In both groups, the serum levels of M-CSF increased significantly as pregnancy progressed. M-CSF levels were relatively higher in twin pregnancy than in singleton pregnancy at 10, 20, and 30 weeks. At 35 and 38 weeks, the M-CSF levels were significantly higher in twin pregnancy than in singleton pregnancy. Elevation of serum M-CSF supports M-CSF production in the placenta. This elevation in twin pregnancy may be related to increased demand of M-CSF in twin pregnancy.     

The 'O-acyl isopeptide method' for the synthesis of difficult sequence-containing peptides: application to the synthesis of Alzheimer's disease-related amyloid beta peptide (Abeta) 1-42.

An efficient 'O-acyl isopeptide method' for the synthesis of difficult sequence-containing peptides was applied successfully to the synthesis of amyloid beta peptide (Abeta) 1-42 via a water-soluble O-acyl isopeptide of Abeta1-42, i.e. '26-O-acyl isoAbeta1-42' (6). This paper describes the detailed synthesis of Abeta1-42 focusing on the importance of resin selection and the analysis of side reactions in the O-acyl isopeptide method. Protected '26-O-acyl isoAbeta1-42' peptide resin was synthesized using 2-chlorotrityl chloride resin with minimum side reactions in comparison with other resins and deprotected crude 26-O-acyl isoAbeta1-42 was easily purified by HPLC due to its relatively good purity and narrow elution with reasonable water solubility. This suggests that only one insertion of the isopeptide structure into the sequence of the 42-residue peptide can suppress the unfavourable nature of its difficult sequence. The migration of O-acyl isopeptide to intact Abeta1-42 under physiological conditions (pH 7.4) via O--N intramolecular acyl migration reaction was very rapid and no other by-product formation was observed while 6 was stable under storage conditions. These results concluded that our strategy not only overcomes the solubility problem in the synthesis of Abeta1-42 and can provide intact Abeta1-42 efficiently, but is also applicable in the synthesis of large difficult sequence-containing peptides at least up to 50 amino acids. This synthesis method would provide a biological evaluation system in Alzheimer's disease research, in which 26-O-acyl isoAbeta1-42 can be stored in a solubilized form before use and then rapidly produces intact Abeta1-42 in situ during biological experiments.     

The synthesis of Δ2-prostaglandins

The effect of prostaglandin F_2α on ovulation and fertilization was studied in rabbits. The number of ovulation was not affected by subcutaneous injection of PGF_2α but the recovery of ova was significantly decreased when PGF_2α was given either at 12 or 16 h after HCG injection and autopsied 24 h latter. The results suggest that exogenous PGF_2α accelerates ovum transport and expels the eggs prematurely from the female tract and does not impair ovulation or the fertilization processes when given to rabbit at 1 mg/kg B.W.     

Synthesis of 11β-3H-prostaglandin E2

11β-³H-Prostaglandin E₂ was synthesized by the stereoselective reduction of the PGD₂ derivative $\text{2}$ using sodium borotritide.