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Eighty-five male Han-Wistar rats were arranged into three groups: CCl4-exposed rats, CCl4 + betaine-exposed rats, and control rats. To see the effect of betaine alone, five rats of the control and of the CCl4 + betaine groups were sacrificed after 7 days, before exposure to CCl4. After that, two of the groups (the CCl4 and CCl4 + betaine groups) were exposed to CCl4 (1 ml/kg per day subcutaneously [SC] for 4 consecutive days), and one of the groups (control group) was given olive oil (1 ml/kg per day SC for 4 consecutive days). At the start of the study (day 0), day 1, day 2, day 3, day 4, and 3 days after the last CCl4 and olive oil injections (day 7), samples of five rats per group were sacrificed, and the livers were taken for chemical analyses and histological examination. Oral betaine, after the acclimation period of a week, increased the number of mitochondria but not mitochondria size (day 0), compared with the case in control rats. Exposure to CCl4 resulted in centrilobular hepatic steatosis, and the administration of betaine significantly reduced this. Morphometric analyses also revealed that the addition of betaine increased the volume density of rough endoplasmic reticulum (RER) in the perinuclear areas of liver cell cytoplasm (day 7). Additionally, the administration of betaine prevented the reduction of Golgi complexes and mitochondrial figures in the cytoplasm observed after the exposures to CCl4. Also, the volume density of mitochondria was smallest in the CCl4-group, but the difference was not statistically significant. The results indicate that oral betaine either improves recovery or reduces the toxic effects of CCl4 on cell organelles in liver cells of male Han-Wistar rats.

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To study vitamin A transport in dairy cows and heifers around parturition, an enzyme immunoassay for bovine retinol binding protein (RBP) was developed and serum levels determined. Serum and liver concentrations of retinol were assayed by HPLC. Four weeks before expected calving the cows and heifers were divided into two groups each, and half of the animals received a protein supplementation during the dry period. The mean serum RBP concentration 4 weeks before calving was 42 mg l-1 for the cows and 44 mg l-1 for the heifers. The serum retinol concentrations were 0.53 mg l-1 for the cows and 0.42 mg l-1 for the heifers, and the liver retinol concentrations 0.30 mg l-1 and 0.13 mg g-1, respectively. In the groups without protein supplementation there was a significant decrease in serum RBP at sampling 1 week before parturition compared to initial values. The measurement of serum RBP may prove useful in assessment of amino acid availability in dairy cows.

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The following blood constituents were measured in the blood and/or serum from healthy and mastitic cows: lipid peroxides (LPO), alkaline phosphatase (AP), lactate dehydrogenase (LDH), urea, glutathione peroxidase (GSH-Px), prostaglandin E2 (PGE2), cyclic AMP (cAMP), electrolytes (Na, K), white blood cells (WBC), haemoglobin (Hb), haematocrit, eosinophils, and electrical conductivity. The levels of these blood variables, excluding GSH-Px, erythrocytes variables and conductivity, were higher in mastitic than in healthy animals. In the blood of the mastitic animals, the AP and LPO levels were 44 and 38% higher than in the healthy animals. The substantial decline in the GSH-Px levels in mastitic animals (P < 0.05) may be related to changes in lipid peroxidation and PG formation and possibly to oxidative stress caused by the host defences. The eosinophil levels were positively correlated with those of PGE2 and cAMP. The possible implications of these findings for the diagnosis of mastitis are discussed.

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The high incidence of pneumonia caused by Pneumocystis carinii in immunosuppressed patients makes it the most important parasite in non-tropical geographical regions. It has recently been shown to be a fungus, but several aspects of this organism are still poorly understood. A major question of clinical relevance is the nature of transmission and, thereby, the related problem of prevention. The mode of P. carinii transmission is thought to be air, but this is based on circumstantial evidence, the transmissive stage has not been identified. We attempted to capture P. carinii by filtration of air in the vicinity of cages containing P. carinii infected Wistar rats. Using nested polymerase chain reaction amplification of the thymidylate synthase gene to demonstrate P. carinii we were able to demonstrate P. carinii DNA on such filters. This strongly supports the suggested mechanism of transmission of Pneumocystis by means of airborne spores and suggests an approach to their isolation and characterization.

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A total of 12 reindeer calves were divided into two groups and the volume densities of liver cytoplasmic structures were counted using TEM. The six animals in group 1 were allowed to graze freely while those in group 2 were kept in pens and fed with lichens. All of them had a negative energy balance and lost weight from November to April, group 1 by 14.9% and group 2 by 24.8%. In spite of the weight losses, no significant changes in volume densities occurred during the winter. Glycogen remained fairly steady and there was no fatty infiltration into the hepatocytes. The very low incidence of Golgi apparatus in the hepatocytes may have been due to the inhibition of protein and lipid synthesis during the winter. These results point to substantial adaptation to extreme environmental conditions. The animals in group 1 apparently moved about more, which may have caused a greater breakdown of glycogen, resulting in turn in a lower volume density of glycogen in January and March.

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Our aim was to study histopathological changes in lung tissue at the light microscopic and ultrastructural level during recovery from immunosuppression and Pneumocystis carinii pneumonia. Male Wistar rats were immunosuppressed by per oral dexamethasone for 12 weeks to induce P. carinii pneumonia, after which dexamethasone was stopped. Recovery was monitored 1, 2 and 4 weeks after cessation of the immunosuppression. In immunosuppressed animals, CD4+ and CD8+ lymphocytes were both decreased in situ. CD8+ lymphocytes increased above control level at week one. Like CD8+ cells, the ED1+ macrophages increased rapidly in situ. This was accompanied by a progressively increasing migration (more transient for lymphocytes) of macrophages into bronchoalveolar fluid, associated with morphological signs of activation and phagocytosis and proliferation of type II pneumocytes. Bronchoalveolar lavage (BAL) fluid tumour necrosis factor-alpha (TNF-alpha) increased from subnormal levels to a 4 week peak, with an inverse correlation between TNF-alpha and cyst count (r = -0.626). Our observations suggest a sequence of changes characterized by an increase in CD4+ cells, accompanied by a more rapid and prolonged recruitment/activation of CD8+ cells, macrophages and type II pneumocytes.

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Pneumocystis carinii pneumonia (PCP) is characterized by the formation of leaky alveoli and a foamy alveolar exudate. To induce PCP, male Wistar rats were immunosuppressed by oral dexamethasone treatment for 12 weeks, during which time all rats developed PCP. Bronchoalveolar lavage fluid (BALF) was analyzed at that time and at 1, 2, and 4 weeks after the cessation of dexamethasone treatment, during which time the rats were recovering from PCP and immunosuppression (and was compared with the BALF obtained from healthy control rats), for type IV collagenase, elastase, cathepsin G, and collagenase activities. Scores for 72-kDa (matrix metalloproteinase type [MMP-2]) and 92-kDa (MMP-9) type IV collagenase-gelatinase activities correlated with those for BALF macrophages (r = 0.58; P < 0.001) and neutrophils (r = 0.66; P < 0.001), respectively, suggesting that they may, in part, be derived from these cells. However, MMP-2 was constitutively expressed and may play a role in normal tissue remodeling. MMP-9 activity was highest in the group with PCP (1.8 +/- 0.37; P > 0.05), with a gradual decline (1.0 +/- 0.48 by week 4; P > 0.05) toward normal (0.67 +/- 0.42) during recovery, which suggests a role for it in tissue-destructive inflammatory events. In rats with PCP the endogenously active collagenase was present at high levels compared with those in healthy controls (2.6 +/- 0.69 versus 0.17 +/- 0.17, respectively; P < 0.01), but they returned to normal by week 4 of recovery (0.42 +/- 0.30; P > 0.05). Collagenase activity showed a correlation with cyst number (r = 0.57; P < 0.001). The BALF of rats with PCP also contained the serine proteinases, which may act as pro-MMP activators. Ultramorphology disclosed increased pinocytotic activities, subepithelial bleb formation, and degeneration and denudation of the basal lamina. These findings suggest that the increased activities of collagenases in BALF caused by the host response against P. carinii might contribute to leaky alveoli.

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The ultrastructural immunolocalization of target antigens recognized by two monoclonal antibodies against Pneumocystis carinii was investigated using both human- and rat-derived organism. Labelling was seen using both cryo electron microscopy and postembedding in Epon after fixation in glutaraldehyde. The antibodies showed different host species- and developmental stage specificity. One of the antibodies reacted with the outer membrane of all developmental stages of human-derived but not rat-derived organisms, whereas the other reacted only with the cyst form. The distribution of the latter was similar to that reported for methenamine silver, a widely used cytochemical stain for the parasite, and reacted with both human- and rat-derived organisms. The antibodies described appear to be useful markers in studies on the differentiation of Pneumocystis carinii.

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Blood serum concentrations of protein, albumin, prealbumin, transferrin, amino acids and urea were measured in 31 healthy cows 0 to 6 weeks before and 3 to 8 weeks after parturition. In comparison to the precalving values the concentrations of albumin, prealbumin and transferrin were all lower after parturition. Alanine, glutamine, leucine, methionine, serine, and urea concentrations were also lower after calving. Multiple correlation analysis between plasma protein and amino acid concentrations reveals that the synthesis of export proteins in liver may be reduced due to limitation in amino acid availability.

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Bone formation by distraction was studied using three different experimental models: (1) Physeal distraction of the sheep radius was performed in 20 animals. (2) Distraction after osteotomy of the radius was carried out in 39 sheep. (3) Mandibular distraction after osteotomy was performed in 17 sheep. Formation of the organic matrix and osteogenesis were studied by radiographic, histologic, and biochemical methods as well as by electron microscopy. The mode of osteogenesis was essentially similar in all of these distraction models. Bone formation was preceded by organization of the collagenous matrix in the distraction area. In the beginning of the distraction, the gap was composed of a heterogeneous cell population, with large polymorphic fibroblast-like cells. The cells in the central part differentiated into fibroblasts, which remained functionally active as long as distraction proceeded. During physeal distraction, bone formed from the epiphyseal and metaphyseal sides as well as from the surrounding perichondrium. Also, in osteotomy distraction of both tubular bone and mandible, bone formed centripetally from the osteotomized bone ends toward the center of the gap. The organic matrix was composed almost solely of Type I collagen in the earliest stages, suggesting that the mode of osteogenesis differs from bone repair by fracture callus. The structure of the distracted segment was mainly lamellar trabecular. Corticalization of the lengthened bone segment occurred gradually after several months.

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Samples from the rumen, reticulum and omasum of 26 reindeer calves were taken during the winter season. Non-specific alkaline and acid phosphatases, cytochrome and amine oxidases as well as succinate, lactate and 3-hydroxybutyrate dehydrogenases were demonstrated in the epithelium histochemically. The phosphatases were usually present in all the epithelial layers, whereas the activities of the other enzymes decreased in the outer layers and could not be demonstrated in the stratum corneum. The activity of alkaline phosphatase seemed to be highest in the reticulum and lowest in the omasum. The reason for the higher activity of this enzyme in epithelial taps in the rumen and omasum and in the reticular and omasal papillae may be the greater need for effective vertical transcellular transport in these regions. There was a tendency for enzymes other than phosphatases to be more active in the rumen than in the other forestomachs, which probably reflects the higher metabolic activity of the ruminal epithelium. No clear differences between early and late winter could be demonstrated.

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Gradual distraction by external fixation was performed one week after osteotomy of the radius in 12 sheep. Bone regeneration in the distraction area was studied by light and electron microscopy. One week after starting the distraction the gap was composed of hematopoietic cells and fibroblasts. The collagen production had already started and it continued actively throughout the distraction period. The collagen produced by the fibroblasts in the central interzone of the gap was organized according to the direction of the distraction. The osteoblasts were lined up along the collagen bundles and osteoid formation was most active around the capillaries. Mineralization started two weeks after the beginning of the distraction. After cessation of the distraction, separate groups of cartilaginous cells were found in some specimens among the newly formed bone. Our findings suggest that osteogenesis as a result of gradual distraction occurs through the whole distraction area with preceding formation of organized collagen matrix. The bone structure in the distracted segment represents an organized lamellar structure at an early stage of the osteogenesis.

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Feline immunodeficiency virus (FIV) was first isolated and identified in 1986. Since then it has been shown to have a worldwide distribution, and the infection generally appears to have reached a state of endemicity. This is the 1st study of FIV-prevalence in Finland. Serum samples of 196 free-roaming cats were tested for antibodies to FIV and FeLV antigens (Feline leukemia virus). With a combined enzyme-linked immunosorbent assay (ELISA), 13 of the cats (6.6%) turned out to be positive for FIV and 2 for FeLV (1.0%). Adult male cats in the capital area of Finland had a FIV prevalence of 24%, a relative proportion 4.7 times higher than that for females.

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Pneumocystis carinii (PC) is an opportunistic pathogen which causes clinical disease in immunocompromised hosts. Three different staining protocols were employed to detect this organism in lung samples of corticosteroid treated voles in order to discover a suitable method for large-scale screening. The procedures employed were: Grocotts methenamine silver (GMS)-stained paraffin sections, toluidine blue O-stained impression smears, and methenamine-silver-stained frozen sections. GMS-stained paraffin sections were relatively easy to interpret and gave more positive results than the other methods. It seemed to be the satisfactory method for large-scale population analyses. An unexpected result was that methylprednisolone treatment did not induce in voles a similarly fatal pneumocystosis infection as occurred in rats. All infections found in voles were mild. This might be due to species-dependent differences in metabolizing methylprednisolone.

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Because of difficulties in in vitro cultivation, the basic Pneumocystis carinii studies have been carried out on animal models, mainly on rodents immunosuppressed by corticosteroids. Commonly used dexamethasone and methylprednisolone procedures were evaluated. The intensity of infection in rats was statistically significantly higher after 9 weeks' immunosuppression with methylprednisolone than with dexamethasone.

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This communication describes the ultrastructure of the ependyma of the bovine area postrema. The polygonal cells possessed an abundant apical array of microvilli but were nonciliated with the exception of occasional cells at the periphery of the area, which had a central kinocilium. The low squamous cells contained moderate numbers of cytoplasmic organelles. The cells do not appear to be typically secretory, but may function in fluid exchange.

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Early autolytic changes in rat heart muscle were quantified using electron microscopical morphometry. Five different time groups with five rats in each group were used. All the rats were anaesthetised, and the hearts of the rats in one group were fixed by perfusion. The other rats were decapitated, and immersion fixation of the hearts performed one minute, five minutes, 10 minutes or 25 minutes after the death of the animals. When compared to perfusion fixation, the first significant changes were seen in the five minute group. The volume densities of myofibrils and glycogen decreased and the volume density of mitochondria increased. These changes were partly due to a shortening of individual sarcomeres. A gradual shortening of the sarcomeres was seen in the one, five and 10 minute groups. Distinct I bands were frequently seen only in the perfusion group. In the mitochondria a decreased matrix density and a partial disorganisation of the cristae were evident in the five minute group. Swelling of individual mitochondria was seen in the 10 minute group, but the increase in size of individual mitochondria was significant in the 25 minute group. After death, morphological changes occur very rapidly in heart muscle cells. This must be taken into consideration when, for example, slaughterhouse material is prepared and interpreted in ultrastructural studies.

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The prophylactic value of mouse monoclonal antibodies to the pig pathogen Haemophilus pleuropneumoniae was studied. Approximately 250 mg of purified mouse monoclonal antibody specific to capsular antigens of H pleuropneumoniae serotype 2 was given IV to five 9-week-old pigs. Five additional pigs from the same litter served as controls. On the following day, all pigs were given a lethal dose (5 x 10(9)) of H pleuropneumoniae serotype 2 into the trachea. Four controls and 1 pig that was given antibodies died within 24 hours. The surviving 5 pigs developed typical signs of pleuropneumonia. After 6 days, the pigs were euthanatized and their respiratory tracts were examined for pathologic changes. All 5 pigs had pathologic changes, but they were less severe in the 4 pigs that had been given antibodies, compared with those in the control pig.

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The expression of a cell proliferation marker, the human transferrin receptor, was studied in ten human gastrointestinal tumors prior to and after implantation under the renal capsule in the mouse (1-6 days). These data were compared to the increase in tumor size in situ, and to the infiltration of inflammatory cells. All tumors studied expressed the transferrin receptor prior to implantation. Forty of 47 implants expressed the receptor, the strongest expression occurring on day 4, accompanied by a reorganization of tumor tissue to a morphology similar to that before implantation. On days 5 and 6 the expression of the transferrin receptor declined. Implants showed maximal increase in size on days 1 and 2, decreased in size on days 3 and 4, and increased again on days 5 and 6. The increase in the size of the implants on days 5 and 6 was accompanied by considerable infiltration of inflammatory cells, and was probably mostly a result of invading host cells and inflammation. If size alone is used as a criterion for tumor proliferation in this subrenal capsule assay, day 4 seems to be the most appropriate for evaluation. This is supported by a strong expression of a proliferation marker, the human transferrin receptor, during this time.

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