De novo protein synthesis by bovine uterine tube (oviduct) epithelial cells changes during co-culture with bull spermatozoa

Polypeptides secreted by uterine tube epithelial cells (UTEC) may facilitate sperm cell capacitation in vivo. This experiment evaluated the effect of sperm-UTEC co-culture on de novo protein synthesis by epithelial cells of the tubal isthmus. Comparisons of the patterns of proteins secreted into medium were made between four culture groups incubated for 24 h in the presence of 35S-methionine: group 1, sperm cells alone; group 2, control UTEC monolayers; group 3, UTEC co-cultured with sperm cells; and group 4, UTEC partitioned by a diffusible membrane from sperm cells during culture. Two-dimensional PAGE followed by fluorography was used to analyze conditioned medium containing secreted proteins from each group. The experiment was replicated four times. Sperm cells alone secreted no detectable proteins, whereas control UTEC monolayers produced a wide array of polypeptides. Sperm cells attached to UTEC in co-culture within minutes, and the resultant protein profile for these UTEC differed markedly from that of the control UTEC. Several new proteins were seen only from co-cultured cells, whereas other protein groups that were present with UTEC alone were absent in the co-culture medium of group 3. The protein pattern expressed by UTEC partitioned from sperm cells (group 4) was intermediate between that of the group 2 controls and that of co-cultured UTEC (group 3). In summary, the attachment of sperm cells to the UTEC during co-culture changed the types and quantities of proteins secreted into the conditioned medium as compared to those of control UTEC monolayers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fine structural changes of the porcine uterine tube epithelium during early and late pregnancy

Ultrastructural changes in the uterine tube (oviduct) of pregnant gilts have been investigated with special reference to the ciliated, secretory, and stromal cells. Tissue from the uterine tube ampulla and infundibulum was taken from 18 gilts at different stages of gestation (days 31, 36, 101, 102, 107, 110, and 112). Cilia were present throughout pregnancy, and deciliation was not apparent at any stage of gestation. The low epithelium of the uterine tube appeared similar to that of the luteal phase of the estrous cycle when corpora lutea were full grown. Prominent features at end of the gestation were numerous fibrous granules and basal bodies, indicating active formation of ciliary precursor organelles. Fibrogranular aggregates were also present in association with the basal bodies. In addition, numerous polyribosomes, mitochondria, and microtubules were encountered in the cytoplasm of ciliated cells at end of the gestation. The appearance of electron-opaque, fibrous granules during late pregnancy probably could be correlated with increasing endogenous levels of plasma estrogen. Intimate morphologic association between fibrous granules and basal bodies indicate that fibrous granules might provide precursor material for the development of cilia or rootlets. Characteristics ultrastructural changes observed in secretory cells during the estrous cycle were not discernible in secretory cells during pregnancy. The secretory cells appeared similar to those of the luteal phase of the estrous cycle. The apocrine secretory cells contained prominent, apical, cytoplasmic projections; pinching-off process of these protrusions was frequently observed during early and term gestation. Extruded nuclei along with other cytoplasmic organelles were also present, lying free in the tubal lumen. The endoplasmic reticulum was predominantly tubular in form. Synthesis, storage, and release of secretory granules were not apparent at early or late pregnancy. It is suggested that progesterone might have an inhibitory effect on the synthesis, storage, and release of secretory granules. Ultrastructural changes in stromal cells were not apparent at any stage of gestation. The stromal cells appeared similar to that of the luteal phase of the estrous cycle.     

Ultrastructural studies of prepubertal porcine uterine tube epithelium

Ultrastructural details of prepubertal porcine uterine tube (oviduct) were studied in normal, growing gilts and compared with observations reported in other species. Tissues from the ampulla region of uterine tube were taken from 6 prepubertal gilts (106 to 139 days old) to determine cytodifferentiation of ciliated and secretory cells. The epithelium consisted of 2 distinctive cells, the ciliated and the secretory cells. Cilia were observed in the uterine tube of prepubertal gilts; however, degeneration of cilia was not observed in the present study. Most prominent observations were the occurrence of fibrous granules in the apical cytoplasm of ciliated cells. These fibrous granules contained electron-dense material and were present near basal bodies. The most unusual feature was the occurrence of procentrioles around a condensation form. These data indicate that ciliated cells are sensitive to estrogen. Intimate morphologic association between fibrous granules and basal bodies indicate that fibrous granules might provide precursor material for the development of cilia and rootlets. The cytoplasm of the secretory cells contained rough endoplasmic reticulum of tubular form and numerous ribosomes. Evidence for synthesis, storage, and release of secretory granules was not apparent. It is suggested that the secretory cells are not sensitive to the low, circulating concentration of plasma estrogen. The ultrastructure of the stromal cells and lymphatic capillary was described for the 1st time. The uterine tube stromal cells were characterized by prominent nucleus and a few cytoplasmic organelles. The lymphatic capillaries were distinguished by the blood capillaries, their much wider lumen, endothelium with an attenuated cytoplasm, absence of basal lamina, and overlapping and interdigitating intercellular junctions. The fine structure of the porcine uterine tube lymphatic capillary generally resembled that of other mammalian species.     

Observation of ultrastructural changes in cultured retinal pigment epithelium following exposure to blue light

BACKGROUND: The retina can be damaged by light even when levels of energy are well below the threshold for thermal damage, and the experimental damage of the retinal pigment epithelium (RPE) may be induced more easily by blue light than by longer wavelengths of visible light. The present study demonstrates the ultrastructural damage produced by exposure to blue light in cultured RPE. METHODS: Long-Evans rats were enucleated 8-10 days after birth for primary culture. One week after seeding, the monolayer culture of RPE cells was exposed to a cool blue light (wavelength = 440 +/- 10 nm) for 36 h (12 h/day, 3 days) at 2.0 mW/cm2. Transmission electron microscopy was used to compare the exposed RPE with the control. The entire experiment was repeated 3 times independently. RESULTS: The cytoplasm of the exposed RPE exhibited degenerative changes, such as large whorls of membrane, lamellar whorls and whorled inclusions. CONCLUSION: The RPE cells can be damaged directly by blue light after excluding the possible influence of phagosomes. This primary culture of RPE can also serve as an in vitro model for the study of light damage to the RPE.     

Cyclic changes in genital organs and vaginal cytology in cynomolgus monkeys (Macaca fascicularis)

Vaginal cytology was evaluated weekly over 12 months in 20 adult female Cynomolgus monkeys (Macaca fascicularis). After sacrifice of the animals the histology of the ovaries, uterus and vagina were studied in different phases of the menstrual cycle. The cytological examination of the vaginal smears showed that the superficial cells increased in number towards the middle of the cycle and the number of intermediate cells declined gradually. Parabasal cells were observed mainly at the beginning of the cycle; they disappeared towards the middle of the menstrual cycle. During the early follicular phase, the cells were moderately separated from each other, and during the second half of the proliferative or follicular phase, the superficial cells appeared clumped together. Leucocytes were usually absent except for at the beginning of the cycle and in the last few days of the late secretory or luteal phase. The maturation index of the vaginal smears can be considered as a tool for distinguishing the different phases of the menstrual cycle. The microscopic examination of the genital organs showed that during the proliferative or follicular phase of the cycle, which corresponds to the development of the ovarian follicles, the uterus showed growth of endometrial glands, stroma and endothelial cell proliferation with capillary sprouts. Shortly after ovulation and parallel to the formation of the corpora lutea, the endometrium enters the secretory or luteal phase, which is characterized by coiling of endometrial glands, glandular secretion and the differentiation of the spiral artery. The most striking changes in the vagina, is the marked basal cell proliferation and thickening of the stratum granulosum during the follicular phase of the menstrual cycle. The histological changes observed in the vagina demonstrated a good correlation with the observation on cytological examination of the smears. The present study demonstrated that the process of angiogenesis in the uterus during the different phases of the menstrual cycle is a multiple phenomenon involving proliferation, maturation and differentiation.     

Retention of functional characteristics by bovine oviduct and uterine epithelia in vitro

The composition of fluids within the bovine oviduct and uterine lumen, important in fertilisation and early embryonic development, is ultimately determined by the transport properties of the epithelial cells which line the lumen. A preparation has therefore been devised to study the role of these cells in oviduct and uterine fluid formation. Pure preparations of epithelial cells, as judged immunocytochemically, were isolated by enzyme digestion, and grown on collagen filters in primary culture. The cells re-establish intercellular junctions to form a confluent epithelial layer. Serial samples from the apical and basal media were analysed for K+, Na+, Ca2+, glucose and lactate. Bovine oviduct epithelial cells maintained gradients of K+ and Ca2+ (apical > basal) for up to 14 days after confluence, while bovine uterine epithelial cells maintained apical > basal gradients of K+. Both types of epithelium exhibited a small transepithelial electrical potential difference and a higher uptake of glucose and production of lactate in the basal, as opposed to apical medium. There were no consistent differences in any of these parameters with the stage of the oestrous cycle at which the cells were removed. The data indicate that bovine oviduct and uterine epithelia may be isolated and grown as polarised layers in primary culture. The preparations will now enable the mechanisms underlying the secretion of ions and non-electrolytes to be determined.     

Effect of cholecystokinin-pancreozymin (CCK-PZ) on glycoprotein secretion from mouse gallbladder epithelium: an ultrastructural and cytochemical study

L-Azetidine-2-carboxylic acid, the naturally occurring lower homologue of L-proline, is incorporated into hemoglobin S (sickle hemoglobin) in vitro. Sickle erythrocytes from patients with sickle cell anemia incubated with L-[3H] azetidine-2-carboxylate synthesized radiolabeled hemoglobin which when isolated from cell lysates, co-chromatographed with hemoglobin S on DEAE-cellulose columns. The alpha/beta ratio of azetidine carboxylate incorporation into the globin chains of sickle hemoglobin was 0.94, which is consistent with the presence of four proline residues in each polypeptide chain. Incorporation of azetidine carboxylate into hot trichloroacetic acid-insoluble material in sickle erythrocytes indicated that the homologue was present in the polypeptide backbone of the globin chains of sickle hemoglobin. Amino acid analysis of the hot trichloroacetic acid-insoluble material from sickle erythrocytes which had been incubated with radiolabeled azetidine carboxylate indicated that 75% of the radioactivity could be accounted for as intact homologue while 20% of the radioactivity co-chromatographed with alanine. These results suggest that azetidine carboxylate is incorporated unaltered into hemoglobin S in addition to being metabolized to alanine in sickle erythrocytes prior to incorporation into protein. The kinetics of thermal precipitation of hemoglobin S (oxygen ligand) into which radioactive azetidine carboxylate or radioactive proline had been incorporated in vitro is identical. This observation, together with the behavior of hemoglobin S and the globin chains from hemoglobin S containing azetidine carboxylate during ion-exchange chromatography, indicates that homologue replacement of prolyl residues does not significantly alter the overall charge or stability of the hemoglobin S tetramer. Azetidine carboxylate did not inhibit uptake of radiolabeled proline by sickle erythrocytes suggesting that the homologue does not adversely affect amino acid transport in these cells.     

Significance of a diastolic notch in the uterine artery flow velocity waveform induced by uterine embolisation in the pregnant ewe

OBJECTIVE: To determine whether water exposure causes middle ear contamination in patients with collar button tympanostomy tubes (TTs). METHOD AND DESIGN: An in vitro model of a human head that contained an auricle, external auditory canal, tympanic membrane with TT, middle ear, eustachian tube, and mastoid cavity was developed. Two electrodes connected to an external ohmmeter resided in the middle ear to detect water entry. The model was tested with 4 types of water exposure: showering, bathing, hair rinsing, and swimming. Statistical analysis was performed by the Fisher exact test. MAIN OUTCOME MEASURES: A positive test result corresponded to water entering the middle ear via the TT, confirmed by a resistance reading of zero on the ohmmeter. A negative test result indicated no change in the initial high resistance reading. RESULTS: No positive test results were obtained for showering (0 of 60 tests), hair rinsing (0 of 60 tests), or head submersion (12.7 cm) in clean tap water (0 of 60 tests). Ten positive test results were obtained for head submersion in soapy water (10 of 97 tests), which was statistically different from clean water (P< or =.007). Swimming pool depths of 30, 45, 60, and 75 cm elicited positive test results in 2 of 16, 3 of 18, 2 of 20, and 11 of 20 tests, respectively. A higher incidence of water entry into the middle ear occurred at depths of more than 60 cm (P< or =.001). No statistical difference between depths of 60 cm or less occurred (P= .88). CONCLUSIONS: Showering, hair rinsing, and head submersion in clean tap water do not promote water entry into the middle ear. Submersion in soapy water increases the probability of water contamination. Pool water infrequently enters the middle ear with head submersion, but the incidence increases with deeper swimming (>60 cm). These data provide further evidence that many water precautions frequently advised in patients with TTs are unnecessary.     

Superfoetation following artificial insemination in a ewe (author's transl)

A case of superfoetation following artifical insemination in a ewe is reported. Estimation of the age of the lambs was based on the maturity of the liver, lung, kidney, brown adipose tissue and skin, showing a difference in gestational age of approximately thirty-six days. The possibility of spontaneous superfoetation as well as that of a second fertilization after artificial insemination is discussed.     

Oral hairy leukoplakia with ultrastructural evidence of Merkel-like cells in human tongue epithelium

The purpose of this investigation was to examine cells in the stratum basale and subjacent lamina propria of oral hairy leukoplakia (OHL) specimens for previously unreported morphological changes. Tongue biopsy specimens were obtained from ten HIV-positive and five healthy male patients and examined by light (LM) and transmission electron microscopy (TEM). Observations made during the investigation revealed the following: (1) LM comparisons of control and OHL specimens indicated that clear cells in the stratum basale ranged from 3 to 7 per 100 basal epithelial cells in healthy tongue mucosa vs. 6 to 15 in OHL. (2) Merkel-like cells were noted in the stratum basale of control biopsies from the dorsal-lateral tongue surface. However, the frequency of observation was so rare as to imply that their presence was an exception rather than the norm. (3) In contrast, Merkel-like cells in OHL specimens were commonly encountered, indicating a possible unique relationship to the pathology. (4) Merkel-like cells associated with OHL lesions were morphologically similar, in all respects, to that of control specimens except for the presence of large, membrane bound, dense granules that ranged in size from 150-300 nm. (5) All biopsies of OHL exhibited evidence of Epstein Barr virus in the stratum corneum and superficial layers of the stratum spinosum. (6) A mild inflammatory infiltrate associated with OHL specimens revealed cytopathic changes in fibroblasts that appeared related to the presence of lymphoid cell types; indicating a possible cytolytic lymphocyte-mediated degradation of antigen altered host cells.     

Bone marrow changes in HIV-positive patients (clinical, cytological, histological, and ultrastructural study of 57 cases)

Individuals with systemic autoimmune diseases develop autoantibodies to nucleosome antigens. For many years, investigators have devoted much effort to precisely mapping epitopes on individual chromatin components. This approach, however, overlooks the existence of determinants that result from multimolecular interactions among nucleosome elements, such as DNA and histones. Anti-nucleosome antibodies can recognize a variety of complex epitopes and are especially prevalent in spontaneous and drug-induced lupus. Using numerous monoclonal anti-nucleosome antibodies obtained from autoimmune mice, we have further characterized these determinants and sequenced the variable region genes of these autoantibodies. I herein review these studies and their implications for the origin of antinuclear autoantibodies.     

Cycloheximide-induced ultrastructural changes in hepatocyte nuclei in partially hepatectomized rats

Nearly all the chromatin in regenerating rat hepatocytes appears to decondensed form 24 h after partial hepatectomy. When cycloheximide (CXM) is administered to partially hepatectomized rats, a marked condensation of chromatin occurs; 4 h after administration the quantity of condensed chromatin present is much higher than that found in the hepatocytes of sham-operated, untreated rats. No segregation or fragmentation of the nucleolus were, however, observed; this shows that the condensation of chromatin is not by itself sufficient to induce the segregation and fragmentation of the nucleolus. The mechanism governing CMX-induced chromatin condensation in regenerating hepatocytes is discussed.     

Microscopic changes in the ligated and resected stumps of the uterine vessels during abdominal hysterectomy (author's transl)

30 stumps of the uterine vessels during abdominal hysterectomy were excised with the ligature 10 to 20 minutes after the original ligature. Inspite of the ligature the most important mechanism of hemostasis is an occlusive thrombosis in the lumen of the vessels. In this process damage of the wall of the vessel and the endothelium is very important.     

The direct effects of circulating lidocaine on uterine blood flow and foetal well-being in the pregnant ewe

We infused lidocaine intravenously into 13 pregnant ewes at a rate sufficient to maintain plasma levels between 2-4 mugm/ml, which are those commonly seen with regional anaesthesia. These levels did not change maternal blood pressure, pulse rate, cardiac output, stroke volume, total peripheral resistance, uterine blood flow or intra-amniotic pressure. Foetal blood pressure and pulse rate remained stable as did both maternal and foetal blood gases and acid-base values.     

The influence of the maternal heart rate on the uterine artery pulsatility index in the pregnant ewe

We investigated the influence of the maternal heart rate on the uterine artery pulsatility index in pregnant ewes. We used an external pacemaker to alter the heart rate of 5 pregnant ewes at 16-17 weeks of pregnancy and examined the effect of changes in the maternal heart rate on the uterine artery flow velocity waveforms and the pulsatility index, as determined by Doppler velocimetry. The uterine artery pulsatility index showed a significant negative correlation with the maternal heart rate. There were no significant changes in other hemodynamic parameters. The maternal heart rate had a significant influence on the uterine artery pulsatility index.     

Proliferation in the rat olfactory epithelium: age-dependent changes

Vertebrate olfactory sensory neurons are replaced continuously throughout life. We studied the effect of age on proliferation in olfactory epithelium in postnatal rats ranging in age from birth (P1) until P333. Using BrdU to label dividing cells, we determined the proliferation density of basal cells, i.e., the number of labeled nuclei/unit length (240 microm) of olfactory epithelium in coronal sections from six different anterior-posterior levels from each animal. A total length of >1 m of olfactory epithelium was counted in each age group. We observed a dramatic decrease of proliferation density from P1 through P333. At P1, proliferation density is 151 cells/mm; it decreases to approximately half at P21 (70 cells/mm), and half again at P40 (37 cells/mm). At P333 the proliferation density was only 8/mm, approximately 5% of that seen at P1. The changes were clearly related to age and not to body weight, because the values were essentially identical for males and females of the same age but of different body weight. Proliferating cells appear in patches that, after P40, become more separated from one another and contain fewer cells. In 6- and 11-month-old rats, 30 and 45% of all units contained no labeled cells. We confirmed the data of others that the olfactory surface area continuously increases with age; we showed that there is a reciprocal relationship between proliferation density and surface area. The proliferating cells provide neurons to sustain growth as well as to replace dying cells.