Immunolocalisation of the uterine secretory proteins uterocalin, uteroferrin and uteroglobin in the mare's uterus and placenta throughout pregnancy.

Previous studies have shown that the equine uterus produces many progesterone-dependent proteins throughout gestation. In particular, uterocalin and uteroferrin are detectable using electrophoresis or blot analyses but information regarding the immunohistochemical placental distribution of these two proteins is rare and information regarding uteroglobin is still lacking. The aim of the present study was to co-immunolocalise these three secretory proteins in the mare's uterus throughout gestation in an effort to understand their functional role in the maintenance of pregnancy. Therefore, endometrial biopsy samples were obtained from 20 pregnant mares between 16 and 309 days of gestation and labelled immunohistochemically for uteroglobin, uteroferrin and uterocalin. Uteroferrin remained detectable in almost every endometrial gland at all stages but with an increase in staining intensity as gestation advanced. The most progesterone-dependent protein, uterocalin, showed variable staining throughout gestation with the most intense labelling in early pregnancy and during the period of endometrial cup reaction. Uteroglobin secretion was only detectable in traces and only in individual glands throughout gestation. The results indicate that uterocalin and uteroferrin, but not uteroglobin, may play important roles in supplying nutrients for the conceptus, thereby contributing to the maintenance of pregnancy. However, further investigations are necessary to understand the role of uteroglobin during gestation.

Histomorphological and immunohistochemical characterization of equine granulosa cell tumours.

Benign (n=33) and malignant metastasizing (n=1) granulosa cell tumours (GCTs) from 34 mares aged 3-21 years, and normal (control) ovaries from nine mares aged 3-10 years, were examined histologically and immunohistochemically (for inhibin alpha, glutathione S-transferase alpha [GSTalpha], c-erbB-2 oncoprotein [cerb], cytokeratin, vimentin, desmin and alpha-actin), the results being related where appropriate to clinical signs and endocrinological data. Availability permitting, serum samples from GCT-affected mares before and several weeks after ovariectomy were examined for the following hormones: oestradiol, progesterone and testosterone (by radioimmunoassay); and inhibin B (by a cross-reactive ELISA). Histological examination revealed that the GCTs were predominantly well differentiated neoplasms. The metastasizing GCT differed immunohistochemically from the benign GCTs in respect of the expression patterns of vimentin, cerb and GSTalpha in the granulosa cells. A notable feature was the presence of Leydig-like cells in mares with stallion-like behaviour or elevated serum testosterone, or both. GSTalpha immunolabelling indicated that the Leydig-like cells were potential producers of steroid hormone. From the immunohistochemical and endocrinological findings it was concluded that GCTs produce abnormally high concentrations of inhibin, which reduce the release of follicle-stimulating hormone, leading to atrophy of the contralateral ovary-a finding in 27 of the mares.

Formaldehyde-free collagen glue in experimental lung gluing.

Because of the well-known limitations of the adhesive strength of fibrin glue, it is imperative to develop a stronger glue with acceptable biocompatibility. This was accomplished by removing the formaldehyde component from gelatin-resorcinol-formaldehyde glue and replacing it by two less toxic aldehydes--pentanedial and ethanedial. To evaluate the adhesive strength of this new glue, GR-DIAL, lung incisions in rabbit hybrids were glued together. Each group (n = 5) was examined histologically after 2 days and 1, 2, and 4 weeks. The glue disintegrated gradually with good bioresorption when the incision was closed with a thin layer of glue. The healing process was favorable, indicating good biocompatibility. Therefore, GR-DIAL glue is capable of enhancing the use of surgical glues in the field of thoracic surgery by enabling surgeons to close larger parenchymal lesions than with fibrin glue.

Lenticular lesions in harbour seals (Phoca vitulina).

Lenticular lesions were found in 21 of 44 mainly younger harbour seals investigated during the epidemic caused by phocine distemper virus (PDV) in 1988/89. Malformations and cataracts varied in quality and degree and were histologically characterized by proliferative and/or degenerative lenticular alterations. Except for two animals, both eyes were affected symmetrically. For that reason, local exogenous causes are improbable. There was very little evidence for infectious agents isolated from the seals investigated (viruses, in particular PDV, bacteria, mycoplasma, parasites). Environmental, dietary and hereditary aspects were also considered, but no final conclusions could be made as to the involvement of these factors in the genesis of lens lesions in harbour seals.

Incidence and morphology of endometrial angiopathies in mares in relationship to age and parity.

The morphology of endometrial blood vessels in uterine biopsy specimens from mares of varying age and reproductive status was examined by light (n = 117) and electron microscopy (n = 13), and additionally after elastase digestion (n = 86). Inflammatory vascular alterations were observed in 20.5% of the specimens. Smaller and larger arterial and venous vessels demonstrated mild to severe degenerative lesions. Unaltered vessels were detected only in maiden mares. Vessels in older maiden mares were frequently affected by angiosclerotic changes, characterized by mild to moderate perivascular and intimal sclerosis. The incidence and severity of angiosis increased with the number of previous pregnancies and with advancing age. Changes in multiparous mares resembled the so-called "pregnancy-sclerosis" of other species, with fraying and disruption of the membrana elastica interna, medial atrophy intimal, medial and adventitial elastosis and fibrosis, and calcification processes within the media. Ultrastructural studies revealed characteristic arterial changes in post-parturient mares, namely, disruption of the membrana elastica interna, as well as activated smooth muscle cells and immature elastic fibres within the intima and inner media, suggesting a pregnancy-induced pathogenesis. Haemodynamic and hormonal alterations during pregnancy and the puerperium possibly induce active vascular remodelling. Cycles of vascular growth during pregnancy and subsequent involution post partum are thought to result in progressive degenerative vascular changes, as seen in multiparous mares. Ageing processes, chronic inflammation and short foaling intervals have to be considered as additional pathogenetic factors. Furthermore, severe angiosis was frequently combined with phlebectasia and lymphangiectasia. This may indicate a reduced ability of the vessels to adapt to the varying demands of uterine circulation, with a decrease of uterine perfusion and lymph drainage. Angiosis in older, multiparous mares might therefore be intimately related to infertility.

Enzootic Mycobacterium avium infection in captive Parma wallabies (Macropus parma) with unusual spinal cord manifestations.

Pathological, microbiological and clinical findings in Parma wallabies (Macropus parma) with an enzootic infection caused by Mycobacterium avium are described. Disorders of locomotion resulted from granulomatous myelitis, meningitis and radiculitis in the lumbosacral spinal cord. Lesions found in a number of internal organs suggested protracted systemic infection due to an alimentary or airborne infection with Mycobacterium avium. This organism was isolated and identified by methods that included microbiological examinations, based on microscopy, culture, in-vitro tests, animal inoculation, polymerase chain reaction and cycle sequencing.

Physiological and pathological expression of intermediate filaments in the equine endometrium.

The aim of this study was to investigate the expression of the intermediate filaments cytokeratin, vimentin and desmin in the equine endometrium by immunohistological techniques. For this purpose, endometrial biopsies of 151 mares were examined to determine physiological cycle patterns and changes resulting from endometriosis. During the physiological cycle epithelial cells and mesenchymal cells express cytokeratin and vimentin, respectively, whilst desmin and vimentin were coexpressed by the smooth muscle cells. Epithelial coexpression of cytokeratin and vimentin was seen in numerous fibrotic glands and in the uterine glands of three mares with pathologically inactive endometria. Three different staining patterns (basal, perinuclear, diffuse) of vimentin were associated with typical morphological alterations of the affected epithelia. In addition, in 14 cases a stromal coexpression of vimentin and desmin was found, indicating an atypical stromal differentiation in inactive endometria of older mares, barren for several years.

Cyclical endometrial steroid hormone receptor expression and proliferation intensity in the mare.

The aims of this study were to investigate the steroid hormone receptor expression and the proliferation intensity during the equine endometrial cycle by immunohistological methods, established for routine examination of formalin-fixed, paraplast-embedded specimens. Endometrial biopsy specimens were obtained during one cycle from 7 mares. In comparison with the blood steroid hormone levels the quantity and distribution of oestrogen (ER) and progesterone receptors (PR) and the proliferation marker Ki-67 antigen expression were investigated. Rising 17beta-oestradiol concentrations in preoestrus induce a synchronous expression of ER, PR and Ki-67 antigen in stromal cells. In the early dioestrus 17beta-oestradiol levels decrease and progesterone levels reach their maxima. This correlates with an intense proliferation activity and the highest hormone receptor expression in epithelial cells. In accordance to the morphological features of asynchronous glandular differentiation in fibrotic areas (endometrosis) their epithelial hormone receptor expression is out of phase.

Nail fragility syndrome and its treatment.

For reasons of clarity, definitions are offered for strength, hardness, flexibility, brittleness and toughness of the nails. Six clinical types of nail fragility are delineated: longitudinal furrows and splitting (onychorrhexis), single longitudinal splitting, multiple crenellated splitting, lamellar splitting (onychoschizia), transverse splitting and nail friability. Changes may be observed in the keratin structure of fragile nails. Nail brittleness is usually 'environmental' in origin, but sometimes may be part of a nail dystrophy. Household daily chores are particularly damaging. Among the acquired general causes, hypochromic anaemia and sideropaenia, arthritic deformities of the distal joints, peripheral vascular impairment and endocrinopathies are the best known. Useful therapeutic approaches are updated. They entail protection with plastic gloves worn over light cotton glove linings, the use of nail hardeners composed of two main types of products: a modified nail varnish that functions as a base coat or a hardener, such as dimethyl urea, which overcomes the objections related to formaldehyde; a systemic drug, biotine, is still useful.

Decreasing B-wave amplitude noted in erg in one type of retinitis pigmentosa.

Electroretinographic studies have been helpful in differentiating human forms of retinitis pigmentosa inherited by different patterns. The prognosis appears to be correlated with the mode of inheritance. The recessively inherited form along with the x-linked and dominantly inherited with reduced penetrance type of inheritance are associated with more severe forms of the disease than is the dominantly inherited with complete penetrance. The underlying biochemical defects have not been identified. Human pathological specimens early in the disease have been almost nonexistent. Animal models have been identified but correlation with human disease has been markedly limited [1]. Electrophysiology and recent technology offer some prospect for differentiating responsible abnormalities and offer a noninvasive evaluation of the disease process and treatment. Three cases of dominantly inherited retinitis pigmentosa with complete penetrance are presented. These show a decreasing B-wave amplitude on successive photopic evaluations using fast random stimuli.

First-order Wiener kernel visually evoked potentials obtained from multiple sclerosis patients.

Visually evoked potentials (VEP) were measured in multiple sclerosis patients with five sizes of reversing check stimuli. The VEPs were obtained using random binary sequence triggered check reversals. The random binary sequence was cross-correlated with scalp potential responses to obtain the estimates of the linear response of the system. For each evaluation a series of five VEPs, one for each of five check sizes, was done. A correlation coefficient was calculated to evaluate check size versus amplitude of the major negative wave. A tabulation was made to compare Snellen chart visual acuity with these correlation coefficients. For the eyes with a Snellen acuity of 20/20, 63% of the VEP amplitudes increased as the check size was decreased with correlation coefficients of -0.6 to -1. For the eyes with a Snellen acuity of 20/70 or less the amplitude decreased with the check size decrease, showing a correlation coefficient of +0.6 to +1 in 45% of the evaluations. The check size giving the largest amplitude was also tabulated with respect to the visual acuity, but did not appear to have as strong a relationship to visual acuity.

Age-related changes in the first order Weiner kernel visual evoked potential.

Visual evoked potentials were obtained from 112 subjects (215 eyes) aged 5 to 79 years with fast random stimuli. The stimulus (the presence or absence of the initiation of a reversal) and response (voltage measurements) were used in calculating the first order Wiener kernels to obtain the visual evoked potentials. Five check sizes were used to stimulate each eye. The implicit time of the major negative wave showed a significant increase with age for each check size used. Regression lines for the implicit time vs age data for male subjects were significantly different from those of females. The y-intercepts were lower and the slope of the regression lines were consistently steeper for males. Thus the rate of increase in implicit time was higher in males. The relationship of the amplitude of the major wave to age did not appear to be linear. The data suggested that a second-order polynomial curve might be an appropriate representation of the relationship.

[The pathogenesis of the "endometritis-pyometra complex" in the female dog]. / Untersuchungen zur Pathogenese des "Endometritis-Pyometra-Komplexes" der Hündin.

Based on investigations in 51 bitches (uteri and ovaries after ovariohysterectomy), suffering from "endometritis-pyometra-complex" (without, n = 38; with, n = 13, hormonal pretreatment), morphologic-functional endometrial characteristics (light microscopy, scanning electron microscopy, enzyme histochemistry, immunohistochemical estrogen receptor analysis) are correlated with ovarian findings, plasma estradiol and progesterone levels as well as microbiological results. The estrous phase of all cycling patients was determined as "diestrus". Plasma estradiol and progesterone concentrations fluctuate within physiological variations. In all cases, not including those, pretreated with progestagen, histological signs of irregular hyperproliferation of uterine glands are obvious, closely related with enzyme histochemical aberrations and atypical endometrial estrogen receptor state. This is interpreted as indicative for prolonged periestrous estrogen induced effects, undergoing different stages of secretory transformation due to the species specific long lasting corpus luteum period. A comparable pathogenesis is supposed deriving from estrogen applications (e. a. after mismating) due to intensified hormonal stimulation and sensibilization of the endometrium, predisposing to ascendant infections by facultative pathogenic bacteria (e. a. E. coli, Streptococcus sp.). Patients without ovarian cyclicity, resulting from progestagen application, exhibit findings, that indicate hormonal imbalances: secretory hypertrophy, irregular regeneration, fibrous atrophy and rigid secretion, resembling those possibly occurring in women after long administration of oral contraceptives.

Endometritis in cattle experimentally induced by Chlamydia psittaci.

On the day of estrus, eight virgin heifers received intrauterine inoculations of yolk sac propagated Chlamydia psittaci strain BovEnd 11/88 isolated from the uterus of a slaughter cow. All heifers developed purulent vaginal discharge which persisted for 3 to 7 weeks. Chlamydiae or chlamydial antigen were detected in vaginal and uterine discharges of infected animals by culture or Capture ELISA, while other bacterial pathogens were not found. In sera of the chlamydia-infected heifers marked increases in antibody titres against the chlamydial genus-specific LPS-antigen were found by ELISA and complement fixation test. Six heifers were artificially inseminated in 5 successive cycles beginning at the first estrus following intrauterine inoculation. In two of the infected heifers spontaneous healing of endometritis occurred after 5 estrus cycles. Only these animals conceived after the 5th breeding, whereas in the remaining four animals a chlamydia-associated chronic endometritis was recognized as the cause of infertility in the 19th and 26th week p.i. at slaughter. Two control heifers which remained clinically normal after intrauterine exposure to sterile yolk sac-suspensions conceived at the 1st and 2nd service, respectively.

Functional disturbances in the endometrium of barren mares: a histological and immunohistological study.

Different types of endometrial maldifferentiation were investigated by histopathological (haematoxylin-eosin (HE), periodic acid Schiff-alcian blue and picro-sirius red staining) and immunohistological (oestrogen and progesterone receptors, Ki-67 antigen, intermediate filaments and laminin) analysis of endometrial biopsy specimens from mares (n=34), most of which had been barren for at least 1 year. Two major features were observed: (i) unequal differentiation, in which two functional stages are visible within a single biopsy sample: areas which are differentiated in accordance with the stage of the ovarian cycle and multiple glandular nonfibrotic foci deviating from the dominant physiological pattern; and (ii) irregular endometrial differentiation, in which there are diffuse abnormalities of functional glandular morphology that cannot be appointed to any physiological endometrial stage. The basic diagnosis was made in HE-stained slides. Special stains and immunohistological analysis confirmed endometrial maldifferentiation concerning at least one, but mostly all, of the parameters investigated. Endometrial maldifferentiation was not related to the age or parity of mares, or to the annual season. The aetiopathogenesis and prognosis of endometrial maldifferentiation are not known. However, the findings of the present study indicate that endometrial maldifferentiation should be included in the routine assessment of endometrial biopsy specimens.

Automated counting of microbial colonies.

The time required for direct counting of colonies on agar plates for estimating population density of viable microorganisms has precluded studies requiring measurement of such a parameter. Bowman, Blume, and Vurek have described a photo-electrical capillary tube scanner which automates the counting. Results obtained with an instrument similar to that of Bowman et al. have been intensively analyzed with respect to precision and accuracy. The sources of "errors" have been ascertained, and the instrument's potentialities and limitations are discussed.

The impact of gelatin-resorcinol glue on aortic tissue: a histomorphologic evaluation.

PURPOSE: Although gelatin-resorcinol-formaldehyde glue has been used to treat acute aortic dissections for some time, concerns about formaldehyde's mutagenicity and carcinogenicity made it imperative to develop a new glue compound. Gelatin-dialdehyde glue was produced by omitting the formaldehyde component and replacing it with two less toxic aldehydes, glutaraldehyde and glyoxal. This study evaluated the histomorphologic effects of the new glue through in vivo use on the aortic tissue of domestic pigs. METHODS: Each animal's infrarenal aorta was glued around an implanted prosthesis. Histomorphologic evaluation was performed after operation after 1 and 4 weeks. RESULTS: The results demonstrated that the clinically observed tanning effect can be attributed primarily to the disintegration of the fiber texture, specifically collagenous, as well as smooth muscle fibers, and to the reciprocal alterations of the proteoglycan interstitial substance in the aortic wall. Macroscopic, microscopic, and electron microscopic analysis of the gluing process revealed an adequate healing process without any morphologically significant difference between formaldehyde and formaldehyde-free gelatin-resorcinol glue. CONCLUSIONS: Gelatin-dialdehyde glue is able to produce the same effects in the area of the aortic wall as the substantially more toxic gelatin-resorcinol-formaldehyde glue and thus could be recommended for clinical trials for treating acute aortic dissections thus far yielding excellent initial results.