To study consequences of cystic duct occlusion, 121 NMRI mice were operated: 40 had cystic duct ligature (L group), 41 had cystic duct ligature and evacuation of bile from the gallbladder (LE group), and 40 were sham-operated (S group). Ten mice from each group were sacrificed at 1 week, 1, 3 and 6 months, respectively. All but 2 mice showed signs of wellbeing during the observation time, and weight gain was the same in the three groups. Distended gallbladder occurred in 14 animals, 7 each in the L and the LE group, more frequent after 3 and 6 months. In the remainder mice the gallbladders were smaller than at the time of operation (p < 0.0001). Fibrosis and adhesions around the gallbladder were frequent in the L and the LE groups, and more pronounced in the latter (p = 0.0001). At microscopy there were signs of inflammation in the gallbladder wall in 16 cases in the L group, 28 in the LE group and 2 in the S group, and the degree of inflammation was more pronounced in the LE group than in the L group (p = 0.002). Eleven mice in the LE group had empyema, and slight intralumininal inflammation was more frequent in the LE group than in the L group (p < 0.0001). In conclusion, despite a high incidence of microscopic inflammation the study gave no indication that occlusion of the cystic duct did serious harm to the mice in this series.

Semen was obtained by vibration-induced ejaculation from 5 spinal injured men once a week for 5 consecutive weeks under standardised conditions. The site of the spinal lesions varied from C5 to Th10. Although in all subjects except one, the total sperm count in the first ejaculate was within normal limits, conventional criteria indicated a high degree of asthenoteratozoospermia in all cases. In subsequent ejaculates there was no major general improvement in motility, vitality or morphology. However, 2 individuals exhibited a marked increase in the proportion of motile sperm in the ejaculate over the next 3 weeks. Flow cytometry of the same sperm samples indicated a high degree of abnormal chromatin condensation and reduced binding of a fluorescent acrosomal marker in the first ejaculates. No improvement in these parameters could be detected with time.Assessment of accessory sex gland function using specific secretory markers indicated that compared to the normal population, the vesicular contribution was markedly reduced in 3 subjects and prostatic contribution in the 2 remaining subjects in the first and subsequent ejaculations. Ejaculate volumes were consistently low in all subjects during the observation period. In contrast, total epididymal secretion was comparable to normal ejaculates. Prostatic contributions to the ejaculate increased significantly over the first 4 weeks.In conclusion, regular vibration-induced ejaculation at weekly intervals could not improve sperm quality in paraplegic men to an acceptable degree for assisted fertilisation to be recommended. Although certain aspects of sperm quality, as judged by conventional criteria, were improved in some cases, flow cytometry revealed persistent chromatin and acrosomal abnormalities.

OBJECTIVE:To determine the extent to which sperm chromatin varies in its degree of condensation in 53 apparently normozoospermic men and in 14 men of proven fertility.METHODS:DNA flow cytometry after staining of spermatozoa with ethidium bromide and mithramycin.RESULTS:Normozoospermic men could be divided into four subgroups according to the fluorescence pattern of their sperm DNA. More than half of the normozoospermic group and all of those of proven fertility shared a similar profile with a narrow symmetrical peak of haploid fluorescence without significant sub-populations. Deviations from this pattern were generally not associated with any major differences in sperm quality.CONCLUSIONS:(a) Current criteria for normozoospermia should be expanded to include additional tests of biochemical integrity. (b) Fertilizing ability appears to be associated with a relatively homogeneous profile of sperm DNA condensation.

DNA flow cytometry of sperm from 100 randomly chosen men undergoing fertility investigation revealed a general association between reduced sperm quality, as judged by conventional parameters, and the appearance of sperm with lower degrees of chromatin condensation in the ejaculate as measured by DNA fluorescence intensity. Chromatin hypocondensation, as measured by increased fluorescence, was manifested to different degrees in different samples. In many cases of more extreme sperm pathology, such as oligoasthenoteratozoospermia (OAT), the whole population of spermatozoa appeared to be affected. Significant numbers of hypercondensed spermatozoa were present in both normozoospermic men and men with different degrees of disturbance in sperm quality. All of the different parameters of sperm quality could be correlated significantly with certain of the flow parameters, although not one in particular could be used to predict deviations from the normal flow profile. In several asthenoteratozoospermic men and a small proportion of men with OAT, the DNA profiles were normal, implying that in these cases the disturbance may not be so fundamental. The presence of leucocytes in the ejaculate was associated with a general increase in the preponderance of hypocondensed subpopulations of spermatozoa in men with OAT as well as in normozoospermic subjects, emphasizing the effect of inflammatory conditions in the reproductive tract on sperm quality.

The binding of a specific, fluorescent acrosomal marker (FITC-labelled peanut lectin) to spermatozoa from men exhibiting differences in sperm quality by conventional criteria was quantitated by flow cytometry. When a standard number of cells was analysed, binding of the lectin diminished and became more variable as the degree of sperm pathology increased. In general, ejaculates with total sperm counts exceeding 120 x 10(6) cells exhibited a stable level of binding within relatively narrow limits. In two normozoospermic men, low levels of acrosomal fluorescence were demonstrated. The significance of these observations with regard to prognosis in assisted fertilization programmes is discussed.