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The cDNA encoding human cystatin C (HCC) was subjected to site-specific substitution of alanine for serine at the position 37, to obtain the Asn35-Lys36-Ser37 sequence that is a signal for asparagine-linked (N-linked) glycosylation of protein in eukaryotes, and was transformed into Pichia pastoris X33. As a result, 1.2 mg/L oligomannosyl HCC with a carbohydrate chain of Man10GlcNAc2 was produced by the Pichia transformant. The oligomannosyl HCC was more stable at the low ionic strength condition of 50 mM potassium phosphate buffer, pH 7.0, than the wild-type. In addition, the oligomannosylation substantially improved the molecular stability of cystatin against an aspartic proteinase, cathepsin D, in which the susceptibility decreased to less than 50% of nonglycosylated one. The anti-rotavirus activity of HCC was substantially enhanced by the site-directed glycosylation using the yeast expression system. A MA-104 cell line was used as a host cell for human rotavirus type-2 Wa strain in this study, to which both the wild-type and oligomannosyl HCCs did not show cytotoxicity at a concentration of 100 μg/mL. More than 80% viability of the host cell infected with 1.0 × 10⁵ PFU/mL of rotavirus was conserved under the condition coexisting with 75 μg/mL of the oligomannosyl HCC, which was 15.2% higher than that of wild-type HCC. Thus, the in vitro anti-rotavirus assay indicated that the supplement of a proper amount of the oligomannosyl HCC could be used as an anti-rotavirus agent.

Simultaneous fetal and maternal ovine placental lactogen (oPL) and prolactin (oPRL) measurements were determined from blood samples collected from six mixed breed (MB; gestation = 141.5 ± .7 d) pregnancies and single Rambouillet (n = 2; Ra gestation = 148 and 149 d) and Finnish Landrace (n = 2; Finn gestation = 141 and 143 d) pregnancies for the purpose of examining a possible relationship between fetal-maternal oPL and oPRL levels in the prepartal period. The MB pregnancies were produced by embryo transfer and consisted of genetically different fetuses co-existing in a common uterine environment. Despite the apparent prematurity of Ra co-twins in MB pregnancies, similar oPL and oPRL patterns during the final 14 d of gestation were observed for both Ra and Finn co-twins. Fetal oPL levels decreased at parturition (range 23 to 90 ng/ml) to approximately 60% of values recorded 8 d prepartum (range 100 to 150 ng/ml) in both fetal siblings. Increases in fetal oPRL concentrations, first observed at approximately 4 d prepartum in all fetuses, reached peak concentrations (40 to 60 ng/ml) at term. Results of this study suggest a similarity in regulation of fetal oPL and oPRL secretion, despite genetic differences for gestation period, in fetuses in the M.B. pregnancy.

Summary.Myoelectrical activity in the uterus of late-pregnant sheep was recorded by the chronic implantation of bipolar electrodes in the myometrium. Three distinct patterns of electrical activity were recognizable: irregular trains of action potentials (>120 sec), long spike bursts (7–120 sec), or short spike bursts (<7 sec) each exhibiting high (>100 μV) or low (50–100 μV) amplitude. The duration of spike activity was expressed as the myoelectrical index (MI), i.e. the fraction of the hour during which a given category of activity is exhibited.The increase in the overall MI which represents daily average values regardless of the class of amplitude from 72 h pre partumup to parturition was significantly (P< 0·05) more pronounced in the uterine horn than in the body of the uterus. During the last 6–7 days of gestation high-amplitude irregular trains and long spike bursts were significantly (P< 0·05) more frequent than these activity patterns with low amplitude. Regardless of the amplitude, the incidence of irregular trains began to decline rapidly 48 h pre partumwith an abrupt cessation following parturition. The frequency of long spike bursts increased 72 h pre partumto reach a maximum which coincided with parturition.

Ethyl ether extracts derived from coffee were tested for in vitro estrogenic and in vivo uterotropic activities. Coffee extracts, unlike tea and cocoa, were found to actively compete with 17ss-estradiol for uterine cytosol binding sites. The biologically active fractions possessed an unique ultraviolet absorbance spectrum that excluded them from containing flavonoid, coumestan, or resorcyclic acid lactone constituents. Coffee extracts administered to immature female mice for 3 d in feeding studies displayed significant (p < 0.05) uterotropic responses, which were similar to results obtained in mice treated with a standard 17ss-estradiol dose. Additional studies in mice disclosed that coffee extracts did not reduce the uterotropic effect normally induced by 17ss-estradiol when administered simultaneously with estradiol. The complete estrogenic effects of coffee constituents, coupled with their failure to inhibit a biological response evoked by estradiol, strongly suggest that coffee contains constituent(s) that are weakly estrogenic.

Ovine embryos collected from two breeds of ewes possessing different gestational periods [Finnish Landrace (Finn), 145.5 ± 1.4 days; Rambouillet (Ra), 150.7 ± 1.3 days] were transferred to common, randomly selected recipients. Fetal plasma samples were collected from mixed-breed (MB, one Finn and one Ra; n= 6) and single-breed (SB, single Finn, n= 3; single Ra, n= 3) chronic fetal preparations during the last trimester. In the MB pregnancy, the Ra sibling had temporal patterns of androstenedione similar to those observed in the Finn co-twin; however, these particular changes were greatly accelerated in comparison to the same day of gestation in SB Ra fetuses. Similarly, temporal patterns of change in unconjugated estrone, estrone sulfate, and 17β-estradiol were accelerated by as much as 4.6 to 5 days in Ra fetuses of MB pregnancies when compared to Ra fetuses in the SB pregnancies (P<0.001), with no difference noted between Ra and Finn co-twins. Maternal progesterone concentrations changed from 8 to 2 ng/ml, androstenedione from 200 to 400 pg/ml, estrone from 65 to 250 pg/ml, and 15-keto-13,14-dihydroprostaglandin F2αfrom 200 pg/ml to 6000 pg/ml over the last 4 or 5 days of gestation. Chronic fetal manipulation accelerated parturition, with mean gestational lengths in MB, SB Finn, and SB Ra pregnancies being 141.5 ± 0.7, 141.7 ± 0.7, and 149.3 ± 0.9 days, respectively. In this study fetal endocrine changes were largely controlled in the MB pregnancy by the fetal co-twin exhibiting a shorter gestational period.