Distribution of Galanin and Galanin Receptor 2 in the Pre-optic Area of the Female Guinea Pig.

This study describes the distribution of galanin (Gal) and galanin receptor 2 (GalR2) in the pre-optic area (POA) of the female guinea pig. Frozen sections were undergone for a routine immunofluorescence labelling. Gal and GalR2 display immunoreactivity in all parts of the pre-optic area. Gal shows reactivity both in perikarya and fibres, whereas GalR2 was observed only in perikarya. Gal- and GalR2-immunoreactive (-ir) perikarya were the most numerous in the medial pre-optic area (MPA) with the highest reactivity in its dorsal part. In the median pre-optic nucleus (MPN) and periventricular pre-optic nucleus (PPN), only single Gal- and GalR2-ir neurons were observed. The highest density of Gal-ir fibres was revealed in the PPN and the lowest in the lateral pre-optic area (LPA). The results of this study indicate that the distribution pattern of Gal containing neurons overlaps well with the distribution pattern of GalR2-positive neurons, especially in the MPA. This may suggest GalR2-dependent activity in this brain region.

Cranial mediastinal liposarcoma in a horse.

A 23-year-old Anglo-Arabian mare was presented with tachypnea, dyspnea, and pitting edema of the ventral thoracic subcutis. On necropsy, a tan to red, friable, irregularly shaped mass (23 × 20 × 18 cm) occupied the cranial mediastinum. Histologically, the mass was classified as a liposarcoma and was composed of short interlacing bundles of spindle-shaped to irregularly rounded cells with discrete, variably sized, clear cytoplasmic vacuoles, which were stained with oil red O in frozen sections of formalin-fixed tissue.

Characterization of the gastric immune response in cheetahs (Acinonyx jubatus) with Helicobacter-associated gastritis.

Captive cheetahs have an unusually severe progressive gastritis that is not present in wild cheetahs infected with the same strains of Helicobacter. This gastritis, when severe, has florid lymphocyte and plasma cell infiltrates in the epithelium and lamina propria with gland destruction, parietal cell loss, and, in some cases, lymphoid follicles. The local gastric immune response was characterized by immunohistochemistry in 21 cheetahs with varying degrees of gastritis. The character of the response was similar among types of gastritis except that cheetahs with severe gastritis had increased numbers (up to 70%) of lamina proprial CD79a+CD21- B cells. CD3+CD4+ T cells were present in the lamina propria, and CD3+CD8α+ T cells were within the glandular epithelium. Lymphoid aggregates had follicular differentiation with a central core of CD79a+/CD45R+ B cells and with an outer zone of CD3+ T cells that expressed both CD4 and CD8 antigens. MHC II antigens were diffusely expressed throughout the glandular and superficial epithelium. No cheetah had evidence of autoantibodies against the gastric mucosa when gastric samples from 30 cheetahs with different degrees of gastritis were incubated with autologous and heterologous serum. These findings indicate that T-cell distribution in cheetahs is qualitatively similar to that in other species infected with Helicobacter but that large numbers of lamina propria activated B cells and plasma cells did distinguish cheetahs with severe gastritis. Further research is needed to determine whether alterations in the Th1:Th2 balance are the cause of this more plasmacytic response in some cheetahs.

MHC class II+ (HLA-DP-like) cells in the cow reproductive tract: II. Immunolocalization of MHC class II+ cells in oviduct and vagina.

The aim of this study was to determine and examine the distribution of major frequency MHC II+ cells in the oviduct and vagina of cows during the oestrous and dioestrus phases. Right oviduct (ampulla, isthmus) and vaginal samples taken from a total of twenty seven multiparous cows were used. Tissue samples were processed to obtain both cryostat and paraffin sections. Sections were stained immunocytochemically using StreptABC method using a specific monoclonal antibody to MHC II+ cell population. Intra-epithelial and subepithelial areas along with lamina propria, muscularis mucosae and serosa of both ampulla and isthmus and intra-epithelial/subepithelial areas and mucosae of vagina were examined for the presence of MHC II+ cells. The density of immune positive cells was determined using a subjective scoring system. MHC II+ cells were demonstrated in all areas examined in both oestrus and dioestrus. In oestrus, the density of MHC II+ cells decreased in subepithelial areas (in between the epithelial cells and the basal membrane) of isthmus, whereas the density of immune positive cells was increased in muscularis mucosae of isthmus (P < 0.05), lamina propria and muscularis mucosae of ampulla (P < 0.05) as well as in the mucosae of vagina (P

Studies of Atlantic salmon (Salmo salar L.) blood, spleen and head kidney leucocytes using specific monoclonal antibodies, immunohistochemistry and flow cytometry.

Monoclonal antibodies (mabs) raised against Atlantic salmon serum IgM (C7G7 and G2H3) and isolated peripheral blood leucocytes (PBL) (E3D9, C4B6 and D8B3) were applied in this study. Using immunoenzymehistochemistry, immunofluorescence and flow cytometry, the distribution of mab+ cells in blood, spleen and head kidney from Atlantic salmon were studied. Immunostaining on cytospin preparations and flow cytometry of isolated PBL showed that the Ig+ cells recognised by C7G7 and G2H3 were mononuclear leucocytes (MNL). The cytospin preparations showed some Ig+ cells with strong cytoplasmic staining, most likely plasma cells. The salmon blood neutrophils were the only E3D9+ cells in cytospin preparations of PBL, and E3D9 recognised about 94% of the defined neutrophil fraction in flow cytometry. The reactivities of C4B6 and D8B3 were to a large degree similar in both immunoenzymehistochemistry and flow cytometry, recognising both MNL and blood neutrophils. Immunofluorescence double staining of PBL with C4B6 and D8B3 showed double staining of all mab+ cells and D8B3 was apparently not able to block the binding of C4B6 to PBL. Immunofluorescence double staining of PBL also revealed more E3D9+ than C4B6+ neutrophils. In immunostaining on cryostat sections of spleen and head kidney, staining of cells was observed with all the mabs, the head kidney generally containing more positive cells than the spleen. Some potential applications for immunological studies using these mabs are suggested.

Use of monoclonal antibodies for immunohistochemical study of bovine spleen on frozen sections.

Many monoclonal antibodies reactive with bovine leukocyte differentiation antigens are now available. Immunohistochemical staining on frozen sections using these monoclonal antibodies permits study of the functional morphology of bovine spleen. This study confirms accepted notions (B and T dependent-zones) and supplies complementary data about the repartition of CD4 and CD8 cells, gamma delta T cells, MHC (Major Histocompatibility Complex) II expression, and macrophages.

[Immunohistology as a reliable and efficient method for the diagnosis of BVDV infections]. / Immunhistologie als zuverlässige und effiziente Methode für die Diagnose von BVDV-Infektionen.

Persistent infection with the Bovine Viral Diarrhea/ Mucosal Disease Virus (BVDV) can be detected using immunohistological methods. The experiments show that the LSAB (Labeled Streptavidin Biotin) peroxidase method is suitable to detect the BVDV in cryostat sections of unfixed tissue. For the diagnostic work up in living animals, skin biopsies give reliable results. In dead animals, organs such as thyroid gland, skin, mucosa of the mouth, esophagus and abomasum are well suitable for immunohistological BVDV detection.

Multifocal myositis associated with Sarcocystis sp in a horse.

Multifocal myositis was diagnosed in a 7-year-old Quarter Horse gelding on the basis of history and findings on physical examination, serum biochemical analysis, electromyography, and microscopic examination of frozen sections of muscle biopsy specimens. Histologic examination of the muscle specimen revealed multifocal accumulations of histiocytes, lymphocytes, and plasma cells, with attendant myofiber degeneration and necrosis. Parasitic cysts with morphologic characteristics of Sarcocystis sp were found in regions of myocyte degeneration and necrosis, and in regions of normal muscle. Based on a tentative diagnosis of Sarcocystis sp-induced myositis, the horse was treated with trimethoprim/sulfamethoxazole and pyrimethamine for 28 days, phenylbutazone for 5 days, and paddock rest for 30 days. At the end of treatment, the horse had gained 35 kg, its appetite had returned to normal, and muscle mass was returning to normal. Sarcocystis fayeri is the only Sarcocystis sp reported in equine muscle in the United States and is rarely associated with acute myositis or muscle atrophy. The development of clinical signs in this horse could have been the result of an underlying immunosuppression or infection with a particularly pathogenic strain or large infective dose of S fayeri.

The accuracy of intraoperative diagnoses based on examination of frozen sections. A prospective comparison with paraffin-embedded sections.

The accuracy of diagnoses based on examination of frozen sections was determined by comparing the results to those obtained by examination of tissues prepared using conventional methods (formalin fixation, paraffin-embedded tissue). One hundred ninety-four specimens were examined using the frozen section technique; 37 were examined to confirm a tentative diagnosis or to document lymph node metastasis and the remainder were examined to diagnose an unknown pathologic process. Of the 194 specimens examined, an accurate, specific diagnosis was obtained in 161 (83%); in 19 (10%), the pathologic process was correctly identified, but a specific diagnosis was not obtained; and in 2 (1%) the diagnosis was deferred. The remaining 12 (6%) were incorrectly diagnosed by the frozen section technique. When the number of specimens in which a specific diagnosis was obtained was combined with the number of specimens in which the pathologic process was correctly identified, the overall accuracy rate of the frozen section technique was 93%. There was no difference in the accuracy of the frozen section technique based on the reason for submission of the sample, source of tissue submitted, or the type of pathologic process (i.e., inflammatory or neoplastic). Of the 12 incorrect diagnoses, 4 (33%) were because of sampling errors and 8 (67%) were caused by interpretation errors.(ABSTRACT TRUNCATED AT 250 WORDS)

Examination of frozen cross sections of cervical spinal intersegments in nine horses with cervical vertebral malformation: lesions associated with spinal cord compression.

Nine horses with clinical and radiographic findings of cervical vertebral malformation that were necropsied and examined using frozen cervical spinal cord cross sections were reviewed. Only cases with actual distortion of the spinal cord due to compression were selected. The goal of the study was to determine the morphologic features responsible for narrowing of the spinal canal and compression of the spinal cord. In individual cases, bony changes are associated with osteochondrosis and osteomyelitis of the dorsal articular facets and osteosclerosis of the dorsal cervical lamina. Soft tissue pathology associated with spinal cord compression included ligamentum flavum hypertrophy, joint capsule swelling and hypertrophy, and synovial cysts. In most cases, a combination of abnormalities was found in horses with spinal cord compression.

Communication of the ulnaris lateralis bursa with the equine elbow joint and evaluation of caudal arthrocentesis.

Elbows from cadaver limbs were evaluated to determine the presence of a communication between the ulnaris lateralis bursa (ULB) and the joint and the extent of the bursa. Thirty-two pairs of joints were studied: 12 pairs were frozen, then transversely sectioned and 20 pairs were injected with methyl methacrylate. The 12 frozen-section pairs revealed a communication between the ULB and the elbow joint in 9/24 joints (37.5%) and a true bursa (absence of communication) in 15/24 joints (62.5%). The mean bursal length in adult horses was 3.8 cm. There was no significant difference in the length of the bursa or presence of bursa-joint communication between the right and left limbs. In the acrylic specimens a communication between the ULB and the joint was found in 19/40 specimens (47.5%). There was no significant correlation between age or sex and frequency of communication. There was a significantly greater prevalence of communications present in Quarter Horse than in non-Quarter Horse (P < 0.05) joints. A communication between the ULB and the joint is not always present, and therefore injection of the elbow joint via the ULB may be unreliable.