Lithium stimulates the first meiotic division in mouse oocytes.

Cumulus enclosed oocytes, cumulus enclosed oocytes denuded of their cumulus and cumulus free oocytes from 21 day old unstimulated mice were cultured for 18 hours in control medium supplemented with lithium chloride, dbcAMP and forskolin at various concentrations. In control medium 66% of the cumulus enclosed oocytes, 93% of the denuded oocytes, and 94% of the cumulus free oocytes resumed meiosis (germinal vesicle breakdown), whereas the levels of polar body formation were 27%, 12% and 39%, respectively. In the presence of lithium significantly more cumulus enclosed oocytes and cumulus free oocytes resumed meiosis and formed a polar body, whereas lithium had no effect on the denuded oocytes. Forskolin and dbcAMP stimulated resumption of meiosis and cumulus expansion in the cumulus enclosed oocytes and inhibited resumption of meiosis in the cumulus free oocytes. Lithium more or less eliminated this inhibitory effect of both forskolin and dbcAMP in the cumulus free oocytes. The results indicate (i) that activation of the cAMP second messenger path in the cumulus cells induces them to synthesize a meiosis inducing substance(s) which stimulates the oocyte to resume meiosis, and (ii) that other second messenger systems than the cAMP pathway, e.g. the phosphatidylinositol cycle, are involved in resumption of meiosis and polar body formation. We conclude that lithium enhances the capability of mouse oocytes for resumption of meiosis and polar body formation.

Direct estimation of volumes in ovarian follicles using the nucleator.

The aim of the study was to evaluate the precision of the nucleator principle with regard to estimation of follicle, oocyte and antral volumes in individual follicles, as well as their average volumes in a group of follicles. Nine isolated murine ovarian follicles embedded in plastic were orientated isotropically, cut into 20 microns serial sections, and stained with hematoxylin. In each follicle the volumes of the follicle, the antrum and the oocyte were estimated in two ways, both unbiased: Firstly, in just one unique section through the follicle, viz. the section which contains the unique sampling point--the nucleolus of the oocyte--using the nucleator principle, and, secondly, in the complete set of follicle sections using the Cavalieri principle. Estimates using the Cavalieri, which are very precise (CEFollicle approximately 0.03, CEOocyte approximately 0.11, CEAntrum approximately 0.16), were interpreted as the "true" values. Therefore, the precision (estimator variation) of the nucleator estimates could be calculated. The nucleator estimates correlated well with the Cavalieri estimates: r(follicle) = 0.95; r(antrum) = 0.88; r(oocyte) = 0.83. The nucleator provides very precise estimates of the volumes of follicle, oocyte and antrum with a minimal amount of effort, since the estimates are derived from measurements in only four directions sampled in a uniform systematic way, the first direction being chosen at random, in only one section through the follicle. Moreover, it is not necessary to know the precise thickness of the section, and finally, the estimates are expressed in correctly dimensioned terms. The estimator variation of the nucleator (CE = 0.28) is of minor importance compared to the biological variation (CV = 0.47) in this realistic sample of follicles.

Anti-cardiolipin antibodies (IgG and IgA) in women with recurrent fetal loss correlate to clinical and serological characteristics of SLE.

AIM OF STUDY: We investigated to which degree IgG, IgA and IgM anti-cardiolipin antibodies (aCL) are associated in recurrent abortion or late fetal death with other signs of autoimmune disease and in particular SLE. MATERIAL AND METHODS: Serological variables typical of SLE and of the anti-phospholipid antibody syndrome were measured once eight to 16 weeks after the last fetal loss in 158 women with recurrent abortion or late fetal death; women with manifest autoimmune rheumatic disease were excluded. RESULTS: (1) Positive values, i.e. above the 99th percentile of reference material, of IgG aCL and IgA aCL were observed in 4% and 7%, respectively, whereas 26% had positive values of IgM aCL. (2) IgG aCL and IgA aCL but not IgM aCL correlated to anti-nuclear antibodies and to anti-double stranded DNA. (3) Anti-double stranded DNA, IgG aCL and IgA aCL but not IgM aCL correlated to previous occurrence of thrombosis. (4) ANA correlated to lower blood platelet concentrations and higher erythrocyte sedimentation rates. CONCLUSIONS: Women with recurrent abortion or late fetal death who have higher but not necessarily abnormally high levels of IgG aCL or IgA aCL constitute a group with increased occurrence of clinical and serological characteristics of SLE. We suggest that these women be kept under surveillance for future development of autoimmune disease especially SLE. The women with high IgM aCL constitute another group without these characteristics.

Total number of particles in a bounded region estimated directly with the nucleator: granulosa cell number in ovarian follicles.

OBJECTIVE: Our objective was to evaluate the precision of the "nucleator" with regard to estimation of the number of granulosa cells in individual follicles and the average number of granulosa cells per follicle in a group of follicles. STUDY DESIGN: Nine murine ovarian follicles embedded in plastic were oriented isotropically with the "orientator" by Mattfeldt, cut into 20 microns serial sections, and stained with hematoxylin. In each follicle the number of granulosa cells was estimated in two ways, both unbiased: (1) with the nucleator principle in the section that contains the nucleolus of the oocyte and (2) in the complete set of follicle sections with the "fractionator" principle. Estimates with the fractionator were interpreted as "true" values. Therefore the precision (CE) of the nucleator estimates could be calculated. RESULTS AND CONCLUSION: The nucleator provides precise estimates of the number of granulosa cells in one complete follicle: CE = 0.12. The amount of effort is minimal, because estimation is based on roughly 50 granulosa cells. The precision of the procedure with regard to estimation of the average number of granulosa cells in a group of follicles depends mainly on the biologic variation (CVbiol = 0.71).

[Qualitative determination of U-pregnanediol-3 alpha-glucuronide in 5 minutes. A clinical trial of a new matrix cushion test]. / Kvalitativ bestemmelse af U-pregnandiol-3 alpha-glukuronid på 5 minutter. Afprøvning af en ny "matrixpudetest".

We have tested an easy and rapid "matrix cushion test" for qualitative determination of U-pregnandiol-3 alpha-glucuronide (U-PGD). The specificity of the test is high, the sensitivity is 4.0 microM, the inter-analysis variation is low as in 96% of the double determinations, agreement was present between the first and second analyses. A positive result on the 21st of the menstrual cycle indicates that ovulation has occurred while this cannot be exluded by a negative or doubtful result. If demonstration of a functioning corpus luteum is also required, the urine should be diluted 1:2 with tap water.

Maturation of mouse oocytes in vitro is influenced by alkalization during their isolation.

Dictyate oocytes were isolated from the ovaries of 22-day-old mice and cultured for 24 h. Isolation and classification of the oocytes were done at a stable pH of 7.45, during gradual increase of pH from 7.45 to 8.3, or at pH 8.3. In cumulus-enclosed oocytes alkalization beyond pH 7.45 resulted in an increased rate of degeneration and decreased rates of germinal vesicle breakdown and polar body formation. In the naked oocytes, only the rate of degeneration was affected. The results stress the importance of a stable pH during the period of isolation, and of a careful classification of the oocytes before culture.

Complete staining of human spermatozoa and immature germ cells combined with phase contrast microscopy.

A method combining Janus green B and Thymol blue stains the anterior part of the head, the nuclear membrane, middle piece, and tail of spermatozoa light green and the nucleus deep purple. The method provides excellent stained preparations for the evaluation of sperm morphology by phase contrast microscopy. It produces significantly less abnormal spermatozoa compared with the Papanicolaou stain.

The precision and accuracy of symphysis--fundus distance measurements during pregnancy.

We studied the precision and accuracy of SF measurements. Five trained obstetricians made three measurements of SF on each of 33 patients in the third trimester. The intraobserver variation varied from 0 cm to 8 cm, and comprised on average 1.5 to 2 cm (95th centiles 2.5 to 4 cm). The interobserver variation varied from 1 to 13 cm, and comprised on average 4 cm (95th centile 8 cm). The accuracy of SF measurements was estimated by comparison with ultrasound-guided measurements, which showed a very high precision. Two of the investigators differed significantly, by -1.56 cm and -1.97 cm, from the other 3.

Emepronium carrageenate: clinical effects and urinary excretion in treatment of female urge incontinence.

Eighteen women (median age 54 years, 36-79) with urinary motor urge (n = 13) or sensory urge (n = 5) incontinence were treated for three 2-week periods with emepronium carrageenate (EC) (Cetiprin Novum) in daily doses of 500 and 1000 mg and placebo. Subjectively the women experienced an increased ability to control micturition, i.e. less urge, during EC treatment. Only mild and mainly anticholinergic side effects were recorded, most frequently dryness of the mouth. As regards side effects, the evaluation of objective effects were complicated by a tendency towards carry-over effect to placebo. Placebo in the first treatment periods were analysed solely; no effects of placebo could be demonstrated. Compared with the reference period, decreases in the total number of micturitions (approx. 20%) and in the number of urge incontinence episodes (approx. 75%) were seen. As compared with placebo, a significant increase (approx. 25%) in the average micturition volume could be demonstrated. The percentage urinary excretion of emepronium decreased with increasing oral intake and with advancing age. Probably, an initial daily dosage of 500 mg EC will do well in younger women (less than 50 years), whereas elder patients may need 1000 mg. Further dosage recommendations are given.

Early prediction of individual serum total estriol curves in pregnancy.

Investigating 60 individual serum total estriol (S-E3) courses from normal pregnancies, we found individual S-E3 courses parallel with the general mean, but with half the standard deviation. Thus one can predict the individual S-E3 course, which may function as a reference for every S-E3 estimation later on. A S-E3 decrease of more than 33 per cent from the individual mean is statistically significant at the 2.5 per cent level.