Histopathological and Histochemical Study of Human Cholecystitis

In our study the following inferences were made. Chronic calculous cholecystitis was seen in the age group of 41 to 50 years. A female preponderance was noted with male to female ratio of 2:3. The most common signs and symptoms were upper abdominal tenderness and right hypochondria pain. The mucosa showed ulceration in 63%, hyperplastic mucosa in 34% and metaplasia in 5%. 22% of cases showed mild (Grade-I) inflammation while 41 % of cases and 27% of cases showed moderate(Grade-2) and severe (Grade-3) inflammation respectively. 44 % of cases showed mild (Grade-I) fibrosis while 22% of cases and 15% of cases showed moderate (Grade-2) and severe (Grade-3) fibrosis. With increase 1n severity of inflammation and fibrosis, the total acid mucin content decreased while the neutral mucin content increased. Similar changes were observed in cases of metaplasia too.

Morphology, histology and histochemistry of the digestive tract of the Banded tilapia, Tilapia sparrmanii (Perciformes: Cichlidae)

This study described anatomical, histological and histochemical features of the mucosal layer of the digestive tract of Tilapia sparrmanii Smith, 1840, an omnivorous freshwater fish endemic to Southern Africa. This species exhibited a short thick oesophagus with long deep longitudinal folds (466.68 ± 16.91 µm), and a thick (173.50 ± 10.92 µm) muscular layer that allow the passage of large food items. The mucosa was lined with stratified secretory epithelium rich in goblet cells that secreted neutral and acid mucins. The stomach was a sac-like structure with simple tubular glands surrounded by connective tissue. The mucosa was lined with simple columnar epithelium and the lamina propria exhibited a well-developed layer of gastric glands that occupied the entire length of the cardio-fundic region. The stomach mucosa consisted of epithelial cells with intense neutral mucin secretion which protects against gastric juice. Neck cells of gastric glands synthesized neutral and acid mucins. The intestine was highly coiled and presented a complex pattern of transversal folds internally (villi). Villi length decreased progressively from the anterior to the posterior intestine (p < 0.0001). Tunica muscularis of the mid-intestine had the thinnest thickness among all parts of the intestine (p < 0.0001). Goblet cells whose numbers increased towards the rectum secreted both acid and neutral mucins. The results indicate structural similarities of T. sparrmaniiGIT with other tilapia species and will be useful for understanding the physiology of the digestive systems as well as functional components of the GIT.

Aging in male Wistar rats associates with changes in intestinal microbiota, gut structure and cholecystokinin-mediated gut-brain axis function

Aging in mammals is characterized by failure of the homeostatic mechanisms that regulate energy balance. Several mechanisms have been proposed such as the presence of a low-grade chronic inflammation in different tissues, as well as leptin and insulin resistance, but the primary alteration is not fully elucidated. The gut microbiota has recently emerged as a key player in a variety of metabolic and neurological disorders. A main concept in this context is the gut-brain axis that refers to alterations in the gut that mediate effects in the central nervous system, including those related with the control of energy balance. Using 16S rRNA analysis, we demonstrate that aged male Wistar rats have increased presence of mucin-degrading and lipopolysaccharide (LPS)-producing bacteria. In addition, old animals exhibit a lower number of neutral mucin secreting goblet cells, and a decrease of tight junctions and adeherens junctions marker proteins, zonula occludens protein-1(ZO-1) and β-catenin, respectively. These data are compatible with a thinner mucus layer and a weaker gut barrier in older animals that likely facilitate LPS leakage. Our data also show that cholecystokinin (CCK) satiating effect is impaired in aged rats, one of the expected effects of increased LPS leakage. In contrast, no overt signs of gut or systemic inflammation are observed. Changes in microbiota in old male Wistar rats present features of situations of increased adiposity, but different from those of obese animals. They could partly explain the increased adiposity and fat deposition in liver and heart observed here.

COMPARISON OF IMMUNOHISTOCHEMICAL MARKER WITH SPECIAL STAINS IN THE PROGNOSIS OF THE DIFFERENT GRADES OF ORAL SQUAMOUS CELL CARCINOMA

Objectives: The aim of the study is to compare the efficiency of Ki-67 with special stains in the stroma of different grades of Oral Squamous Cell Carcinoma (OSCC) and to evaluate the influence of these changes in predicting the prognosis of these tumors. Materials and Methods: A total of 30 cases of different grades of Oral Squamous Cell Carcinoma and 6 cases of control were sections and stained with Picrosirus red (PR), combined Alcian blue-periodic acid Schiff reagent (AB-PAS) and Immunohistochemical (IHC) marker Ki-67. Results: Collagen fibre nature using PR stain and proliferative activity of malignant epithelial cells using IHC marker Ki-67 was found to be statistically significant. Mucin presence using AB-PAS was statistically insignificant. Conclusion: Prognosis in different grades of oral squamous cell carcinoma can be accessed by change in collagen fibres birefringence, as the tumour progresses there is change from mature to immature collagen. Ki-67 is a good proliferative marker and shows that there is positive correlation with histological grading of oral squamous cell carcinoma. Presence of acidic or neutral mucin in OSCC needed to be further studied. For assessing the prognosis in different grades of OSCC, special stains can also be used. Keywords: Collagen, Mucin, Squamous cell carcinoma, Ki-67.

Mucosubstances in the porcine gastrointestinal tract: Fixation, staining and quantification

Mucins are of great interest in intestinal research and histochemical methods are often employed to identify them. Since it is in the nature of mucins that they are “hard to hold onto” once they come into contact with water, a frequently used medium in histochemistry, there are a number of challenges that may decrease diagnostic accuracy. As the outcome of methods published for microscopic detection of mucosubstances proved to be unsatisfactory in our hands, the aim was the establishment of a reliable and reproducible protocol. Tissue samples were available from pig feeding experiments. In the present study, we focus on a fixation / staining procedure without making comparisons between differently fed pigs. Several fixation and staining procedures were evaluated for their use in semiautomatic quantification and quality assessment of different mucus fractions simultaneous on one tissue section. Cryostat sectioning, subsequent fixation steps with heat, ethanol and modified Bouin’s solution, followed by triple staining with high iron diamine, alcian blue and periodic acid-Schiff turned out to be the best method to identify sulfomucin, sialomucin and neutral mucin simultaneous on one tissue section. This methodology resulted in very good morphology of goblet cells with intact mucin containing vesicles within the cells, which was comparable to ultrastructural electron microscopical observations. Semiautomatic quantification of different mucins was possible. In conclusion, reliable mucus quantification and assessment of mucus quality requires strictly tested procedures. According to our experience, the most important aim after cryosectioning is fast fixation of the mucosubstances, which requires a combination of different fixation steps.

Investigating the Suitability of a Laboratory Mouse Model to Study the Pathogenesis of Abortifacient Campylobacter jejuni

The aim of this study was to assess whether pregnant mice represent a useful model to study the reproductive pathology of Campylobacter jejuni IA3902 using the end point of positive microbial culture of the organism from the fetoplacental unit. Pregnant BALB/c and CD-1 mice (14 days’ gestation) were inoculated orally and intraperitoneally (IP) with 1 × 109 colony-forming units/ml of C. jejuni IA3902. The organism was recovered by microbial culture from the fetoplacental unit in 10 of 10 BALB/c and 10 of 10 CD-1 IP-inoculated pregnant mice and in 29% (2/7) BALB/c and 38% (3/8) CD-1 orally inoculated pregnant mice. Gross reproductive lesions included necrosuppurative placentitis, fetal resorption, intrauterine fetal death, stillborn pups (dead neonates), and multifocal hepatitis. Histological changes consisted of locally extensive neutrophilic and necrotizing placentitis with intralesional bacterial colonies of C. jejuni, ulcerative endometritis, random multifocal hepatitis, and rare cholecystitis. Immunohistochemistry for the major outer membrane protein of C. jejuni revealed moderate to large numbers of the organism at the periphery of the placental discs, within trophoblasts and extracellularly, with invasion into the placental disc largely via the vascular network. The organism is trophic for neutral mucin, iron, and L-fucose within the murine placenta. C. jejuni IA3902 has affinity for the murine reproductive tract, specifically the fetoplacental unit, where it results in a necrotizing placentitis with positive microbial recovery after both IP and oral challenge in BALB/c and CD-1 pregnant mice.

A comparative anatomical, histological and histochemical study of small intestine in Kestrel (Falco tunniculus) and white eared bulbul (Picnonotic leucotis) according to their food type

This study was conducted on 30 birds (15 birds for each type) divided as 10 birds for each part of study. Anatomical part revealed that the small intestine in both birds kestrel (Falco tinniculus) and white eared bulbul (Picnonotic leucotis) formed from 3 segments; duodenum, jejunum and ileum with no clear demarcation line between them. In kestrel the Meckel's diverticulum appeared as small projection to separate between jejunum and ileum. Both ratio of intestinal length to body length and of intestinal weight to body weight was higher in bulbul than those in kestrel. Histological study showed that the wall of all three parts of small intestine was composed of the same histological layers; these are mucosa, submucosa, muscularis and serosa. There was almost similarity in structure of these tunics but significant differences in several Histomorphometric measurements of each tunica. Goblet cells were more abundant in all parts of small intestine of bulbul than those in kestrel and there was a gradual increasing in the number of these cells toward the end of intestine of both birds. Histochemical part of this study appeared that in villi and crypts of all small intestinal segments of both birds the goblet cells secrete neutral mucin in nature because it showed negative reaction to Alcian blue stain and positive to PAS stain.

Food restriction beginning at lactation interferes with the cellular dynamics of the mucosa and colonic myenteric innervation in adult rats

The effects of food restriction (FR) on the morphoquantitative aspects of the wall and myenteric neurons of the proximal colon in adult rats were analysed. FR was imposed by duplication of the experimental brood size in relation to the control brood during lactation. The FR group received a 50% reduction of food from weaning until 90 days of age. Samples of the colon underwent histological processing to morphometrically analyze the crypts, muscularis mucosae, tunica mucosa, and muscularis externa. We determined the number of goblet cells and serotoninergic enteroendocrine cells, and morphoquantitatively studied the myenteric neuronal population. FR caused hypertrophy in the tunica mucosa, increase in crypt depth and in the muscular layer of the mucosa, a decrease in the thickness of the tunica muscularis and in the number of goblet cells and an increase in serotoninergic cells. A higher neuronal density in the ganglia and a reduction of the cell profile area were observed in the FR group. FR imposed since lactation led to hypertrophy of the tunica mucosa, a reduction of neutral mucin production, atrophy of the tunica muscularis, and an increase in the survival neuronal in adult rats, attributable to an increase in the number of serotoninergic enteroendocrine cells in mucosa.

Histomorphology Spectrum of Gall Bladder Pathology in Cholecystectomy Specimens with Clinical Diagnosis of Chronic Cholecystitis

Introduction: Chronic cholecystitis is a common disease in surgery where majority of cholecystectomy are performed. This study was carried out to see histomorphological pattern of gallbladder diseases, staining pattern with various stains and prevalence of carcinoma. Methods: This prospective study was conducted in the department of Pathology, B.P. Koirala Institute of Health Sciences, Dharan, Nepal from January to December 2008. A minimum of fifty random samples were included in study. Clinical details and pathological data were retrieved from case sheets and patient’s history. All specimens were fixed in 10% formalin. Three sections each from neck, body and fundus were taken. Tissues were processed in an automated tissue processor and paraffin blocks made. The tissue sections were examined under light microscope after hematoxylin-eosin, neutral mucin, sulfated mucin and sialomucin stain. Results: Chronic cholecystitis without cholelithiasis constituted majority of 25(50.0%) cases with M: F of 1: 7.1. The gastric and intestinal metaplasias were present in 33.0 % and 8.0% of the cases respectively. The neutral, sulfated and sailomucin were positive in 35.0 %, 21.0 %, 1.0 % and 34.0 %, 21.0 %, 1.0% cases respectively in gastric and intestinal metaplasia. Both gastric and intestinal metaplasia showed significant p-value on sulfated mucin. Conclusions: The increase risk factors for developing of chronic cholecystitis were seen in female gender (86%). The gastric and intestinal metaplasia showed positivity more with neutral followed by sulfated mucin. Keywords: chronic cholecystitis; metaplasia; mucins.

Multiple Hepatic Peribiliary Cysts in a Young Pig

Histopathologic features of hepatic peribiliary cysts were described in a young slaughtered pig. The animal was an apparently healthy 6–month-old pig of mixed breed. Macroscopically, all lobes of the liver contained numerous cysts of varying size containing serous fluid in all lobes. Histopathologically, the cysts were located mainly around the large bile duct and in the connective tissue of the portal tracts. Within serial sections, these cysts were assumed to be solitary or multilocular, but they were separated from the bile duct. The cysts were lined by a single layer of columnar, cuboidal, and flattened epithelial cells. Occasionally, goblet cells were observed. The epithelial cells were stained with periodic acid—Schiff/alcian blue and high-iron diamine/alcian blue, indicating the presence of neutral mucin, sialomucin, and sulfomucin. Grimalius' method revealed the presence of endocrine cells in the lining epithelium. There was no bile pigment in the cysts by the Hall method.