Ultrastructural studies on the pancreatic acini in duck (Anas boscas) and pigeon (Columba livia)

The aim of this research work was to study the histological structure of the pancreatic acini by transmission electron microscope in two avian species, duck and pigeon. The specimens were collected and processed for electron microscopic study. The results showed that the acini of the two avian species were two types; the first one was an electron dense and the second one an electron lucent. The light acinar cells were larger in size than the dark cells. These cells contained centrally located ovoid nuclei with prominent nucleoli and abundant euchromatin. The cytoplasm was electron lucent, with many rough endoplasmic reticulum, polymorphic mitochondria. Numerous zymogen granules were distributed in the basal part and around the nucleus, so these cells considered active cells. The dark acinar cells were characterized by an electron dense cytoplasm. The most prominent cell organelle in these cells were the zymogen granules that appeared in different sizes while other organelles as mitochondria, and rough endoplasmic reticulum were inconspicuous or few, so these cells were considered as inactive cells. The nucleus with indented nuclear membrane located centrally with prominent nucleoli and abundant heterochromatin. Prominent intercellular spaces between the individual acinar cells, as well as well-developed basement membrane separating the electron dense cells and the lumen contained the secretion between acinar cells. It could be concluded that the acinar cells in ducks and pigeons were divided into two types, that is, light and dark acinar cells which mainly attributed to the activity of these cells.     

Exocrine pancreatic atrophy in German Shepherd Dogs and Rough-coated Collies: an end result of lymphocytic pancreatitis

Previously published studies of the pathology of canine exocrine pancreatic insufficiency (EPI) have been based on morphological findings during the clinical phase of the disease, when atrophy of acinar parenchyma occurs. Recently, low serum trypsinlike immunoreactivity (TLI) concentration has been shown to precede clinical signs, making it possible to diagnose EPI prior to onset of the clinical disease. This study presents histological and ultrastructural findings of pancreatic biopsies from 11 German Shepherd Dogs and 2 Rough-coated Collies with subclinical EPI (SEPI). These findings were compared with those from dogs with clinical EPI (n = 11) and healthy control dogs (n = 5). Biopsied tissue from dogs with SEPI typically contained both normal and atrophied acinar parenchyma. The most significant finding was the marked lymphocytic infiltration, which was most prevalent at the border zone of affected and nonaffected parenchyma but had spread into the normal acinar tissue. Numerous intraacinar lymphocytes were found. Most of the lymphocytes were positive by immunostaining for CD3. In more advanced stages of destruction, the findings were characteristic of pancreatic acinar atrophy. In the atrophied parenchyma, the inflammatory reaction, if present, was less prominent. Ultrastructural changes were in accordance with those of the histological study showing infiltration of lymphocytes both in affected acini and in acini that revealed no obvious ultrastructural changes. Progressive degenerative changes of acinar cells were considered a nonspecific finding. Apoptotic death of acinar cells was occasionally found. The inflammatory reaction was clearly shown to precede the pancreatic acinar atrophy, and the findings suggested that lymphocytic pancreatitis leads to atrophy of the pancreas. The possibility that EPI is an immune-mediated disease in German Shepherd Dogs and Rough-coated Collies is discussed.     

Normal ultrastructure and histochemical characteristics of canine lacrimal glands

Lacrimal glands of 12 dogs free of ocular disease were examined to determine the normal structure of these glands. The glands consisted of tubuloacinar cells that ultrastructurally and histochemically were of a single type of secretory cell in the tubules and possibly 3 types of secretory cells in the acini. The tubular epithelium contained homogenous electron-dense granules that stained as neutral glycoconjugates (periodic acid-Schiff positive and Alcian blue and high iron diamine negative). The predominant acinar cells contained granules of lesser electron density than those of the tubules, and stained as sialomucin (Alcian blue [pH 2.5] and periodic acid-Schiff-positive, and high iron diamine-negative). A second type of acinar cell was in peripheral lobules that ultrastructurally and histochemically appeared like lipid granules (positive with oil red O and osmium tetroxide). Ultrastructurally, a third type of acinar granule was finely granular, electron-lucent, and frequently coalesced. It was not readily apparent whether the latter was an artifact, a stage in the maturation of the sialomucin granules, or a third type of acinar granule. Individual acinar cells usually had a predominance of 1 granule type, but greater than 1 granule type could be found in some cells. The basal surfaces of the acinar, tubular, and ductal cells were incompletely ensheathed by myoepithelial cells. Plasma cells, lymphocytes, mast cells, endothelial cells, fat cells, and Schwann cells composed the cellular elements of the interstitium. Lymphocytes, mast cells, and nerve endings also were found in the parenchyma.     

Lysosomal storage disease caused by Sida carpinifolia poisoning in goats

A neurologic disease characterized by ataxia, hypermetria, hyperesthesia, and muscle tremors of the head and neck was observed for 2 years in a flock of 28 Anglo-Nubian and Saanen goats on a farm with 5 ha of pasture. Six newborns died during the first week of life, and five abortions were recorded. The predominant plant in the pasture was Sida carpinifolia. The disease was reproduced experimentally in two goats by administration of this plant. Three goats with spontaneous disease and the two experimental animals were euthanatized and necropsied. No significant gross lesions were observed. Fragments of several organs, including the central nervous system, were processed for histopathology. Small fragments of the cerebellar cortex, liver, and pancreas of two spontaneously poisoned goats and two experimentally poisoned goats were processed for electron microscopy. Multiple cytoplasm vacuoles in hepatocytes, acinar pancreatic cells, and neurons, especially Purkinje cells, were the most striking microscopic lesions in the five animals. Ultrastructural changes included membrane-bound vacuoles in hepatocytes, Kupffer cells, acinar pancreatic cells, Purkinje cells, and the small neurons of the granular cell layer of the cerebellum. Paraffin-embedded sections of the cerebellum and pancreas were submitted for lectin histochemical analysis. The vacuoles in different cerebellar and acinar pancreatic cells reacted strongly to the following lectins: Concanavalia ensiformis, Triticum vulgaris, and succinylated Triticum vulgaris. The pattern of staining, analyzed in Purkinje cells and acinar pancreatic cells coincides with results reported for both swainsonine toxicosis and inherited mannosidosis.     

Expression of claudin-5 in canine pancreatic acinar cell carcinoma - An immunohistochemical study

Claudin-5 is an endothelium-specific tight junction protein. The aim of the present study was to detect the expression pattern of this molecule in intact pancreatic tissues and in well-differentiated and poorly differentiated pancreatic acinar cell carcinomas from dogs by the use of cross-reactive humanised anticlaudin-5 antibody. The necropsy samples taken from dogs included 10 nonneoplastic pancreatic tissues, 10 well-differentiated pancreatic acinar cell carcinomas, 10 poorly differentiated pancreatic acinar cell carcinomas, 5 intrahepatic metastases of well-differentiated and 5 intrahepatic metastases of poorly differentiated acinar cell carcinomas. A strong lateral membrane claudin-5 positivity was detected in exocrine cells in all intact pancreas samples. The endocrine cells of the islets of Langerhans and the epithelial cells of the ducts were negative for claudin-5. The endothelial cells of vessels and lymphatic channels in the stroma of the intact pancreas showed strong membrane positivity for this claudin. All well-differentiated exocrine pancreas carcinomas and all poorly-differentiated pancreatic acinar cell carcinoma samples showed a diffuse loss of claudin-5 expression. The claudin-5-positive peritumoural vessels and lymphatic channels facilitated the detection of vascular invasion of the claudin-5-negative cancer cells. In liver metastasis samples, the pancreatic carcinomas were negative for claudin-5. It seems that the loss of expression of claudin-5 may lead to carcinogenesis in canine exocrine pancreatic cells.     

不同周龄SPF级SD大鼠胰腺自发性病变的组织学观察

研究不同周龄SPF级SD大鼠胰腺自发性病变的种类及其病变发生率,为药物安全性评价提供背景资料。收集3年安评试验中11、19、31周龄试验对照组大鼠胰腺制作病理切片,光学显微镜下观察SD大鼠胰腺病变的种类及组织形态学特点,并统计其病变发生率。结果显示,大鼠胰腺主要出现了以下病变:①单核细胞浸润:11周龄大鼠该病变的总体发生率为0.6%,其中雌性为0,雄性为1.25%;19周龄大鼠该病变的总体发生率为1.0%,其中雌性为1.0%,雄性为1.0%;31周龄大鼠该病变的总体发生率为1.0%,其中雌性为0,雄性为1.96%。②腺泡细胞空泡变性:11周龄大鼠未观察到该病变的发生;19周龄大鼠该病变的总体发生率为1.0%,其中雌性为0,雄性为2.0%;31周龄大鼠该病变的总体发生率为3%,其中雌性为2.1%,雄性为3.9%。③腺泡细胞萎缩、腺管增生:11周龄大鼠未观察到该病变的发生;19周龄大鼠该病变的总体发生率为0.5%,其中雌性为0,雄性为1.0%;31周龄大鼠该病变的总体发生率为3.0%,其中雌性为1.0%,雄性为4.9%。结果表明,SPF级大鼠胰腺可发生单核细胞浸润、腺泡细胞空泡变性、腺泡细胞萎缩及腺管增生等自发性病变,且病变发生率可随着年龄的增加而增加。     

Histo-ontogenetic study of the parotid salivary gland of Indian buffalo

The present study was aimed at elucidating the histogenesis of parotid gland of buffalo. The study was carried out on buffalo foetuses (n = 36), during different stages of prenatal life. The foetuses were categorised into three groups based on their curved crown rump length (CVRL). The primordial anlage of parotid salivary gland was evident at 40th day of development whereas the primary ducts, in the form of cords, were first observed at 81st day of prenatal life. The capsule formation as well as the lobulation of the gland was initiated at 127th day. At 141st day, the duct system of gland was completed. The terminal tubules attained the structure of acini at 167th day. The myoepithelial cells first appeared as flattened basal cells initially around the developing acinar cells at 167th day. The typical compound tubulo-acinar nature of the gland was first observed at 185th day. Purely serous acinar cells were seen from 185th day onwards. The micrometrical studies revealed that the mean diameter of acinar cells, intercalated ducts, striated ducts and large ducts increased with the advancement of age. The serous acinar cells were devoid of acidic as well as neutral mucopolysaccharides in prenatal age groups; however, large ducts with goblet cells exhibited positive reaction. Combined PAS-AB method revealed mixed reaction in acinar cells as well as in large ducts. Fine lipid droplets were observed in intralobular as well as interlobular connective tissue; however, phospholipids were observed in the cell membrane of secretory cells and ducts.     

A pathological study of the salivary glands of rabid dogs in the Philippines

Rabies is a zoonotic disease caused by the rabies virus. While the salivary glands are important as exit and propagation sites for the rabies virus, the mechanisms of rabies excretion remain unclear. Here, we investigated the histopathology of the salivary glands of rabid dogs and analyzed the mechanism of excretion into the oral cavity. Mandibular and parotid glands of 22 rabid dogs and three control dogs were used. Mild to moderate non-suppurative sialadenitis was observed in the mandibular glands of 19 of the 22 dogs, characterized by loss of acinar epithelium and infiltration by lymphoplasmacytic cells. Viral antigens were detected in the mucous acinar epithelium, ganglion neurons and myoepithelium. Acinar epithelium and lymphocytes were positive for anti-caspase-3 antibodies and TUNEL staining. In contrast, no notable findings were observed in the ductal epithelial cells and serous demilune. In the parotid gland, the acinar cells, myoepithelium and ductal epithelium all tested negative. These findings confirmed the path through which the rabies virus descends along the facial nerve after proliferation in the brain to reach the ganglion neurons of the mandibular gland, subsequently traveling to the acinar epithelium via the salivary gland myoepithelium. Furthermore, the observation that nerve endings passing through the myoepithelium were absent from the ductal system suggested that viral proliferation and cytotoxicity could not occur there, ensuring that secretions containing the virus are efficiently excreted into the oral cavity.     

A freeze-etching study on experimental murine mastitis.

Streptococcal mastitis was produced experimentally in mice inoculated by the intramammary route; freeze-etched preparations from the affected mammary glands were studied by electron microscopy. The inoculated cocci were seen free in the acinar lumen, within luminal phagocytes and within cells of the epithelium. No significant pathological changes were noted in the junctional complexes between secretory epithelial cells. The results were comparable to those obtained by ultrathin sectioning and indicated that, while cocci can transfer from the acinar lumen into the substance of the epithelium and towards a subepithelial location, the junctional complexes between epithelial cells present a potential barrier to movement through the intercellular spaces.     

Experimental encephalomyocarditis virus infection in Mongolian gerbils (Meriones unguiculatus)

Two strains of Mongolian gerbils (Meriones unguiculatus), Tumble Brook (TUM) and Japan Medical Science (JMS), were intraperitoneally inoculated with the D variant of encephalomyocarditis virus (EMC-D) and killed 3 days later. Mortality was significantly higher in females than in males. Evidence of viral replication was detected in the heart of both strains and in the pancreas of the TUM strain. Histopathological alterations were found in the heart and pancreas. Heart lesions involved foci of necrosis with inflammatory cell infiltration and calcification in both strains. Pancreatic lesions were restricted to the exocrine glands; islets of Langerhans were rarely and secondarily involved in the extensive destruction of exocrine glands. Severe acinar cell necrosis with marked inflammatory edema was conspicuous in TUM, whereas only slight acinar cell involvement was detected in JMS gerbils. Immunoperoxidase staining showed viral antigens in intracytoplasmic vacuoles in damaged acinar cells.     

Spontaneous Adenosquamous Carcinoma with Rapid Growth and EMT-like Changes in the Mammary Gland of a Young Adult Female BALB/c Mouse

This study histopathologically and immunohistochemically investigated a spontaneously occurring single mass subcutaneously located in the left lower abdomen of a female BALB/cAJcl−nu/+ mouse at 10 weeks of age. The mass was about 20 × 15 × 10 mm in size after formalin fixation; nevertheless, it was not detected by clinical observations at 9 weeks of age. H&E staining revealed the tumor origin was epithelial and probably arose from the mammary gland, and the tumor cells demonstrated a squamous, acinar or polyhedral/basal pattern. A cell kinetics analysis revealed that many of the tumor cells of the squamous, acinar or polyhedral/basal component were positive for PCNA and cyclin D1, although there were a few of TUNEL-positive tumor cells in all of the components. An epithelial/mesenchymal analysis demonstrated that most of the tumor cells of the squamous and acinar components contained keratin and E-cadherin; however, most of the tumor cells of the polyhedral/basal component were less or very weakly positive for these markers. The tumor cells of the squamous component were negative for vimentin and SMA; however, many of the tumor cells of the polyhedral/basal component exhibited vimentin. In addition, expression of SMA was confirmed in some tumor cells of the acinar and basal components. Based on the microscopic and immunohistochemical characterizations, the tumor was diagnosed to be adenosquamous carcinoma that originated from the mammary gland with rapid growth, and the tumor cells demonstrated epithelial-mesenchymal transition-like changes.     

Histopathological changes in the pancreas due to decreased pancreatic blood flow in a canine tachycardia-induced cardiomyopathy model

In dogs, pancreatic acinar cell injury is thought to be caused by decreased pancreatic blood flow due to heart failure. In previous our report, it demonstrated that decreased heart function causes a significant decrease in pancreatic blood flow in heart failure dog model caused by rapid ventricular pacing (RVP). However, the types of histopathological changes remain unclear. We aimed to verify the types of histopathological changes occurring in the pancreatic tissue due to decreased heart function. After RVP for 4 weeks, atrophy of pancreatic acinar cells, characterized by a decrease in zymogen granules, was observed in all areas of the pancreas. In conclusion, the result of this study suggests that attention should be paid to ischemia/hypoperfusion injury in the pancreas.     

Juvenile acinar atrophy of the pancreas of a dog.

A dog with juvenile acinar atrophy of the pancreas had degranulation and loss of the acinar cells. Transitional features from normal to degranulation were seen. Electron microscopically, the atrophic pancreas mainly consisted of epithelial cells of duct system, islet cells and multivesicular cells.     

Post-natal maturation of acinar cells of the guinea pig pancreas: an ultrastructural morphometric study

The morphological maturation of the acinar cells of the guinea pig pancreas during post-natal development was characterized morphometrically by determining the intracytoplasmic accumulation of rough endoplasmic reticulum (RER) and zymogen granules. The following results were obtained for the period analysed, i.e., from 2 to 70 days of post-natal life: (a) the acinar cell volume increased by 210% (P < 0.01); (b) the mostly cisternal RER occupied more than 30% of the cytoplasm at any age studied and their total volume and surface in the cell were increased by 300 and 534% (P < 0.01), respectively; (c) maturation in the morphological pattern of the RER was observed; (d) the mean number of zymogen granules per cell increased from 261 at 2 days to 422 at 70 days (P < 0.01), while their mean diameter increased from 0.52 to 0.94 micron (P < 0.01) during the same period; (e) these increases in granule number and size were responsible for a 500% (P < 0.01) increase in total volume from 2 to 70 days and for a 304% increase (P < 0.01) in total surface from 2 to 35 days; (f) the RER and the zymogen granules together occupied 44, 54, 55 and 57% of the cytoplasm at 2, 14, 35 and 70 days of age, respectively. We conclude that although the pancreatic acinar cells of the guinea pig are morphologically well differentiated at 2 days of age, with the cytoplasm already showing a large amount of RER and zymogen granules, they are still immature. Morphological maturation of the acinar cell occurs during the first months of post-natal life and is characterized by a substantial gain in cell volume and intracytoplasmic accumulation of RER and zymogen granules, which significantly increase of both their absolute volume and total surface, with a higher growth rate being observed during the period from 2 to 14 days of post-natal life.     

The effect of pentaghrelin on amylase release from the rat and porcine dispersed pancreatic acinar cells in vitro

Ghrelin-28 was found to inhibit the pancreatic enzyme output in rats, although the effect of pentaghrelin has not been studied. The effect of ghrelin on pig exocrine pancreas remains largely unknown. The aim of the present study was: (1) to establish a model of porcine dispersed pancreatic acinar cells in vitro and compare it with an existing rat model, and (2) to investigate the effect of pentaghrelin on amylase release from the rat and pig pancreatic acini. The rat and porcine acinar cell preparations released amylase in response to cholecystokinin octapeptide (CCK-8) stimulation in a dose-related manner. Pentaghrelin hardly inhibited the amylase release in rat preparations (maximum inhibition with 10^− 9 M pentaghrelin). It stimulated amylase release in porcine preparations, however, no dose response was found in a range of doses between 10^− 10 and 10^− 7 M. Concluding, pentaghrelin may stimulate amylase release from porcine acinar cells through as yet unknown mechanisms.     

Cellular immunolocalization of gastric and pancreatic lipase in various tissues obtained from dogs

OBJECTIVE: To determine cellular immunolocalization of canine gastric lipase (cGL) and canine pancreatic lipase (cPL) in various tissues obtained from clinically healthy dogs. SAMPLE POPULATION: Samples of 38 tissues collected from 2 climically healthy dogs. PROCEDURES: The cGL and cPL were purified from gastric and pancreatic tissue, respectively, obtained from dogs. Antisera against both proteins were developed, using rabbits, and polyclonal antibodies were purified by use of affinity chromatography. Various tissues were collected from 2 healthy dogs. Primary antibodies were used to evaluate histologic specificity. Replicate sections from the collected tissues were immunolabeled for cGL and cPL and examined by use of light microscopy. RESULTS: Mucous neck cells and mucous pit cells of gastric glands had positive labeling for cGL, whereas other tissues did not immunoreact with cGL. Pancreatic acinar cells had positive labeling for cPL, whereas other tissues did not immunoreact with cPL. CONCLUSIONS AND CLINICAL RELEVANCE: We concluded that cGL and cPL are exclusively expressed in gastric glands and pancreatic acinar cells, respectively. Also, evidence for cross-immunoreactivity with other lipases or related proteins expressed by other tissues was not found for either protein. Analysis of these data suggests that gastric lipase is a specific marker for gastric glands and that pancreatic lipase is a specific marker for pancreatic acinar cells. These markers may have clinical use in the diagnosis of gastric and exocrine pancreatic disorders, respectively.     

Cellular and humoral immune responses in atrophic lymphocytic pancreatitis in German shepherd dogs and rough-coated collies

The most common cause for the clinical signs of exocrine pancreatic insufficiency (EPI) in dogs is pancreatic acinar atrophy (PAA). In the subclinical phase of EPI, before total atrophy occurs, exocrine pancreas is affected by infiltrative lymphocytic inflammation, which gradually leads to selective destruction and atrophy of the acinar tissue.Here, we analyzed the role of cell-mediated and humoral immune mechanisms in the pathogenesis of atrophic lymphocytic pancreatitis in German shepherd dogs and rough-coated collies. Pancreas biopsies and serum samples were obtained from 12 dogs with subclinical EPI (SEPI), 13 dogs with clinical EPI and 13 healthy control dogs.Immunohistochemical analysis showed that, in the subclinical phase, the majority of the infiltrating lymphocytes were T-cells with an almost equal number of CD4+ 'T-helper' and CD8+ 'cytotoxic' T-lymphocytes. The distribution of the two lymphocyte subsets was different. Typically, the CD4+ cells were present in large cellular infiltrates in the affected parenchyma, and the scattered CD8+ cells had infiltrated both the affected and the normal parenchyma. In sections where destruction of acinar parenchyma was present, the CD8+ T-cells were predominant. In cases of marked T-cell infiltration, CD79+ B-lymphocytes and plasma cells, and lysozyme-positive macrophages were also detected. Lymphoid follicle germinal centers with a majority of cells staining positively for CD79 were found. The lymphocytic infiltration in the totally atrophic tissue of dogs with clinical EPI was less prominent. Indirect immunofluorescence staining showed serum antibodies reacting weakly with pancreatic acinar cells in five out of nine dogs with subclinical and three out of 10 dogs with clinical EPI, but not in the control dogs.The results suggest that the tissue destruction is largely T-cell-mediated, although the presence of numerous B-lymphocytes and pancreas-specific antibodies in the sera of some dogs indicate that humoral mechanisms are also involved. In conclusion, this study suggests that the atrophic lymphocytic pancreatitis in German shepherds and rough-coated collies is an autoimmune disease.     

Adenoviral pancreatitis in guinea fowl (Numida meleagris)

Necrotizing pancreatitis was observed in 2-week-old Guinea fowl submitted for necropsy and histopathology. Intranuclear inclusion bodies seen histologically in acinar epithelium were examined by electron microscopy and found to contain viruses resembling adenovirus. Adenovirus was isolated in embryonated eggs from the pancreata of affected birds. The adenovirus isolated was not neutralized by chicken antisera developed against 10 known serotypes of group 1 avian adenoviruses.