Cytologic examination of bronchoalveolar lavage fluid (BALF), including phenotypic analysis of lymphocytes, was performed on 32 Standardbreds with poor race performance and endoscopic examination findings characteristic of inflammatory airway disease (IAD). Nucleated cell counts in BALF from IAD-affected horses were higher than those in control horses; the cytologic profile of BALF in affected horses included mixed inflammation, characterized by mild neutrophilia, lymphocytosis, and monocytosis. Eosinophil and mast cell counts were not higher in the IAD-affected group, compared with those in the control group; however, 4 IAD-affected horses had marked eosinophilia (24.7 +/- 4.8% SEM) in BALF. Phenotypic analysis of lymphocytes in BALF obtained from IAD-affected horses revealed a low proportion of CD4-positive cells and B cells, compared with those in the control group; these findings may have been representative of a greater proportion of non-B, non-T cells (null cells) in horses with IAD. The cytologic profile of BALF obtained from horses with IAD differed from that in horses affected with chronic obstructive pulmonary disease, suggesting that the pathogenesis of inflammation in horses with IAD may differ from that of chronic obstructive pulmonary disease.

Thirty-eight tracheobronchial aspirates (TBA) were collected from twenty 1 to 6-month-old foals, which were free of clinical signs of respiratory tract or other infectious disease. We collected TBA from 9 of the foals 3 times when they were approximately 8, 16, and 24 weeks old. Aspirates were examined cytologically after staining with modified Wright-Giemsa, Gram, toluidine blue, and prussian blue stains. Aerobic bacterial culturing was performed on all aspirates. Of the 20 initial TBA, 4 (20%) were normal cytologically on the basis of previously defined criteria for TBA from clinically normal horses, 6 (30%) had a high percentage of eosinophils (> 5%), 8 (40%) were classified as indicative of subacute inflammation, and 2 (10%) were classified as indicative of acute inflammation. Nine (45%) were positive for mast cells and none were positive for hemosiderin-laden macrophages (hemosiderophages). Of the 9 foals from which samples were collected at 16 and 24 weeks of age, results were similar, except for an increase in the number of TBA classified as indicative of chronic inflammation (33% and 22% respectively) and the number positive for hemosiderophages (33% and 88%, respectively). One TBA was considered nondiagnostic because of pharyngeal contamination. Culturing of 12 of the 37 aspirates (32%) yielded a potential microbial pathogen. Only 2 were positive cultures from the same foal. The following organisms were isolated: beta-hemolytic Streptococci spp (4), Actinobacillus/Pasteurella spp (4), Rhodococcus equi (2), unidentified nonenteric Gram-negative rod (1), and Escherichia coli (1). Thirty-four of the 37 aspirates (92%) yielded light growth of various organisms considered to be nonpathogenic and normal inhabitants of the upper respiratory tract. It was concluded that the presence of inflammatory cells, eosinophils, and mast cells in the tracheobronchial aspirates from clinically normal foals is a common finding. These cytologic findings were consistent in the samples collected from foals at 8, 16, and 24 weeks of age. It was also concluded that bacteria with recognized pathogenicity can be isolated from TBA from clinically normal foals and were most frequently isolated from 1- to 2-month-old foals or those with cytologic evidence of inflammation, even in the absence of clinical signs of respiratory tract disease.

Urinary bladder tumors in cats, unlike in dogs, usually appear outside of the trigonal region as localization. These tumors are confused withurinary tract infections associated with hematuria in a clinical sense. Cytological examination and ultrasound (USG) imaging techniques arevery valuable, but histopathological approach is the golden key. In this case neutered male, mixed, 17-year-old cat was brought to the clinicwith non-obstructive hematuria attacks. A superficial hypoechoic mass located at the apex of the urinary bladder was detected in the USG imagingtechnique. No discernible findings were found by cytological examination. After removal of the suspected area, the biopsy specimen waspresented to the pathology department for histopathological examination. Histopathological examination revealed transitional cells coveringentire surface of the mucosal epithelium and showing growth into the lumen, and they were characterized by mild anisocytosis and anisokaryosis.The patient was diagnosed as in-situ non-papillary, non-infiltrating type of transitional cell carcinoma (in-situ carcioma). These tumorsare quite rare and have better prognosis. The condition of the patient was completely resolved without medical treatment in the postoperativeperiod. When this case report was prepared, 6 months after the operation, there was no recurrence in the patient. Contrary to infiltrative urinarybladder tumors, in-situ carcinoma could be completely cured by surgically, therefore the case was found worthy to be presented.

A one and half year old heifer was presented with a hard mass hanging from brisket region. The mass was oval shaped, 10 x15 cm in size, pedunculated with the base at the level of carpal joint. Cytological examination revealed melanocytes with the presence of numerous blackish melanin pigments in the cytoplasm. Histopathological examination revealed that the tumor was comprised of heavily pigmented spindle shaped cells, arranged in sheets with a large nuclei and abundant granular black pigment in the cytoplasm. Following surgical removal of the mass, the animal recovered with no recurrence. Based on clinical examination, cytology and histopathological findings, the growth was diagnosed as cutaneous melanoma.

Objective-To evaluate time-dependent alterations in gene expression of chemokines in bronchial epithelium of recurrent airway obstruction (RAO)-affected horses and whether alterations resulted from increases in gene expression of interleukin (IL)-17 in cells isolated from bronchoalveolar lavage fluid (BALF). Animals-8 RAO-susceptible horses and 9 control horses. Procedure-In 2 experiments, both groups of horses were evaluated after being maintained on pasture and after being stabled and fed dusty hay for 1, 14, 35, and 49 days (experiment 1) or 14 and 28 days (experiment 2). In experiment 1, gene expression of IL-8, chemokine (C-X-C motif) ligand 1 (CXCL1), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), and Toll-like receptor 4 (TLR4) in epithelium and IL-8, IL-17, and TLR4 in BALF cells was measured. In experiment 2, bronchial biopsy specimens were evaluated for IL-8 immunoreactivity. Results-In RAO-susceptible horses after 14 days of challenge exposure, there was a 3- and 10-fold increase in gene expression of IL-8 for epithelial and BALF cells and an increase in IL-8 immunoreactivity in epithelial cells. Challenge exposure failed to alter gene expression of CXCL1, GM-CSF, G-CSF, and TLR4 in epithelial cells of any horses at any time point. During challenge exposure, gene expression of BALF cell IL-17 was downregulated in control horses (day 1) and upregulated in RAO-affected horses (day 35). Conclusions and Clinical Relevance-Epithelial-derived IL-8 may promote airway neutrophilia, but the inciting stimulus is unlikely to be IL-17 because upregulation of this gene is subsequent to that of IL-8 in epithelial cells.

Objective-To determine via histologic examination and scintigraphy the effect of focused extracorporeal shock wave therapy (ESWT) on normal bone and the bone-ligament interface in horses. Animals-6 horses without lameness. Procedure-Origins of the suspensory ligament at the metacarpus (35-mm probe depth) and fourth metatarsal bone (5-mm probe depth) were treated twice (days 0 and 16) with 2,000 shocks (energy flux density, 0.15 mJ/mm2). One forelimb and 1 hind limb were randomly treated, and the contralateral limbs served as nontreated controls. Bone scans were performed on days -1 (before ESWT), 3, 16, and 19. Histomorphologic studies of control and treated tissues were performed on day 30. Results-ESWT significantly increased the number of osteoblasts but caused no damage to associated soft tissue structures and did not induce cortical microfractures. A significant correlation between osteoblast numbers and radiopharmaceutical uptake was noticed on lateral views of the hind limb on days 3 and 16 and on caudal views of the forelimb on day 3. Conclusions and Clinical Relevance-Results suggested that ESWT has the potential to increase osteoblast numbers in horses. The correlation between increased osteoblast numbers and radiopharmaceutical uptake 3 days and 16 days after the first ESWT suggested that stimulation of osteogenesis occurred soon after ESWT. No damage to bone or the bone-ligament interface should occur at the settings used in this study, and ESWT can therefore be administered safely in horses.

Although it has been reported that hormones or chemicals, which increse in intracellular cAMP, produced Mg2+ release from the heart, it is not well characterized whether a specific Mg2+ exchanger is involved in cAMP-induced Mg2+ efflux in themammalian hearts In this work, we studied the relationship between the increase in intracellular cAMP and ion transport system on Mg2+ regulation in the perfused rat heart and isolated myocytes. The Mg2+ content in the perfusate and supernatant were measured by atomic absorption spectrophotometer. The addition of membrane permeable cAMP analogue to the perfused hearts and myocytes. cAMP-induced Mg2+ efflux was ingibited by H7, benzamil or imipramine in the perfused hearts and myocytes, but not by EIPA. We confirmed that a significant Mg2+ efflux was induced by an increase in intracellular cAMP in the hearts and myocytes. The cAMP-induced increase of Mg2+ efflux in the hearts may be involved in ion transport system(Na+-Ca2+ and Na+-Mg2+ exchanger)

The development of uterus in fetuses on 60, 90 and 120 days of gastation and neonates of Korean native goats was investigated by scanning electron microscopy. The results were summarized as follows; 1. in the 60-day-old fetuses, the short microvilli were sporadically observed on the luminal surface of the endometrium. 2. In the 90-day-old fetuses, the mucosal folds, polygonal microridges, numerous microvilli, flower-like-buds, and domeshpaed or crateriform area were also observed on the luminal surface of endometrium. 3. In the 120-day-old fetuse, the primordial caruncles(nodules) of the endometrium were developed conspicuously and long microvilli were developed densely. 4. In the neonates, thecaruncles and microvilli of the endometrium were more developed than those of 120-day-old fetuses.

In the present study we have analyzed the characteristics and distribution of the mu-opioid receptor(MOR) by raising anti-peptide antisera to the C-terminal peptide of MOR. The antisera against MOR was produced in New Zealand White rabbit against 15 residue corresponding to amino acids, 384-398of the cloned rat MOR. The antigenic peptide was synthesized using an Applied Biosystems 432 solid-phase peptide aynthesizer. The specificity and identification fo the antisera were tested by analysisi fo transfected cells, epitope mapping and immunohistochemical method. COS-7 cells electroporated with MOR cDNA were used to evaluate the characteristics and subcellular distribution of MOR.MOR immunoreactivity was prodominent in the plasmalemma and subcellular compartments such as encoplasmicreticulum, Golgi apparatus and vesicle like structure. Furthermore, both tissue sections and transfected cell lines could be immunostained with these antisera and the immunoreactivity ws abolished when anti-MOR sera were preincubated with the peptide against which they wer raised. Based on epitope mapping analysis, all antisera appeared to have a similar epitope, which ws detemined to be within the last amino acid,391-398. Moreover, immunohistochemistry showed that MOR immunoreactivity was ovserved in many brain areas including cerebral cortex, striatum, hippocampus, locus coeruleus and the superficial laminae of the dorsal horn. These stained spinal cord and brain aras showed themirrored pattern observed in autoradiographic studies of mu-opioid binding as well as a pattern similar to that seen by in situ hybridization for MOR. Thus, several lines of evidence support the conclusion that the antisera produced in the present study most likely recognize mu-opioid receptor. These results suggest that MOR antisera may be utilized as useful tool to analyze the physiological and pharmacological studies for mu-opioid receptor in the future

Alpha toxin of S aureus has cytolytic activity respectively. This antigen has been received the most attention since it is a major virulence factor in pathogenesis of staphylococcal mastitis. Thus, alpha toxin has been focused as potential candidate ofvaccine tominimize mastitis in cows. the purpose of this study was to develop a simple, efficient production and purification methods of sufficient amount of alpha toxin antigen from S aureus. Alpha toxin production measured by hemolytic activity was the highest at 18 hrs postinoculation in yeast extract culture medium supplemented with thiamine, nicotinic acid and casamino acid. Alpha toxin was purifed by ammonium sulfate precipitation (65%) and ultrafiltration. Molecular weight of the toxin was 33 kDa in the analysis with SDS-PAGE. Conclusionally, when alpha toxin was included in the vaccine, the optimal harvest time of alpha toxin was at 18 hrs after inoculation in yeast extract medium supplemented with thiamine and nicotinic acid.