Alcian Blue-periodic acid-Schiff Research Articles

Interleukin-13 maintains the stemness of conjunctival epithelial cell cultures prepared from human limbal explants.

To use human limbal explants as an alternative source for generating conjunctival epithelium and to determine the effect of interleukin-13 (IL-13) on goblet cell number, mucin expression, and stemness. Human limbal explants prepared from 17 corneoscleral rims were cultured with or without IL-13 (IL-13+ and IL-13-, respectively) and followed up to passage 2 (primary culture [P0]–P2). Cells were characterized by alcian blue/periodic acid–Schiff (AB/PAS) staining (goblet cells); immunofluorescent staining for p63α (progenitor cells), Ki-67 (proliferation), MUC5AC (mucin, goblet cells), and keratin 7 (K7, conjunctival epithelial and goblet cells); and by quantitative real-time polymerase chain reaction for expression of the p63α (TP63), MUC5AC, MUC4 (conjunctival mucins), K3, K12 (corneal epithelial cells), and K7 genes. Clonogenic ability was determined by colony-forming efficiency (CFE) assay. Using limbal explants, we generated epithelium with conjunctival phenotype and high viability in P0, P1, and P2 cultures under IL-13+ and IL-13- conditions, i.e., epithelium with strong K7 positivity, high K7 and MUC4 expression and the presence of goblet cells (AB/PAS and MUC5AC positivity; MUC5AC expression). p63α positivity was similar in IL-13+ and IL-13- cultures and was decreased in P2 cultures; however, there was increased TP63 expression in the presence of IL-13 (especially in the P1 cultures). Similarly, IL-13 increased proliferative activity in P1 cultures and significantly promoted P0 and P1 culture CFE. IL-13 did not increase goblet cell number in the P0–P2 cultures, nor did it influence MUC5AC and MUC4 expression. By harvesting unattached cells on day 1 of P1 we obtained goblet cell rich subpopulation showing AB/PAS, MUC5AC, and K7 positivity, but with no growth potential. In conclusion, limbal explants were successfully used to develop conjunctival epithelium with the presence of putative stem and goblet cells and with the ability to preserve the stemness of P0 and P1 cultures under IL-13 influence.

Srolo Bzhtang, a traditional Tibetan medicine formula, inhibits cigarette smoke induced airway inflammation and muc5ac hypersecretion via suppressing IL-13/STAT6 signaling pathway in rats

Ethnopharmacological relevanceSrolo Bzhtang (SBT), a traditional Tibetan medicine formula, was composed of three herbs, Solms-Laubachia eurycarpa, Bergenia purpurascens, Glycyrrhiza uralensis, and one lac, and was first documented in the ancient Tibetan medical work Four Medical Tantras (rGyud-bzhi) in the eighth century AD. It has been widely used to treat lung “phlegm-heat” syndromes such as chronic bronchitis and chronic obstructive pulmonary disease (COPD). ObjectiveThe aim of this study was to evaluate the potential influences of aqueous extract of SBT on airway inflammation and mucus secretion and to reveal the underlying mechanism in a rat model of cigarette smoke (CS)-induced chronic bronchitis (CB). Materials and methodsSixty male Sprague-Dawley rats were randomly divided to six groups: control (room air exposure), model (CS exposure), DEX (CS exposure and 0.2 mg/kg/day dexamethasone), and three SBT (CS exposure and 1.67, 2.50, and 3.34 g/kg/day SBT) groups. DEX and the three doses of SBT were administered by oral gavage every day for eight weeks. Pathological changes and mucus expression in the lung tissue were determined by hematoxylin and eosin (H&E), Alcian blue-periodic acid-Schiff (AB-PAS) and immunohistochemical staining. The levels of cytokines in bronchoalveolar lavage fluid (BALF) were assessed by ELISA. Western blot analysis and qRT-PCR were performed to explore the effects of SBT on the expression of IL-13, STAT6 and MUC5AC. ResultsPretreatment with SBT attenuated the TNF-α, IL-8, IL-13 expression levels in BALF and the inflammatory cell infiltration in bronchial walls and peribronchial lung tissue. SBT exhibited a dose-dependent downregulation of MUC5AC expression as assessed by AB-PAS and immunohistochemical staining. The protein and mRNA levels of IL-13, STAT6/p-STAT6 and MUC5AC were also downregulated by SBT preconditioning. ConclusionThese results for the first time demonstrated that SBT exhibited protective effects on CS-induced airway inflammation and MUC5AC hypersecretion, which might be related to the downregulation of the IL-13/STAT6 signaling pathway.

Mucin staining is of limited value in addition to basic immunohistochemical analyses in the diagnostics of non-small cell lung cancer

Accurate diagnosis of histological type is important for therapy selection in lung cancer. Immunohistochemical (IHC) and histochemical stains are often used to complement morphology for definite diagnosis and are incorporated in the WHO classification. Our main aim was to compare different mucin stains and assess their value in relation to common IHC analyses in lung cancer diagnostics. Using tissue microarrays from 657 surgically treated primary lung cancers, we evaluated the mucin stains periodic acid-Schiff with diastase (PASD), alcian blue–periodic acid-Schiff (ABPAS) and mucicarmine, and compared with the IHC markers p40, p63, cytokeratin 5, thyroid transcription factor 1 (TTF-1), napsin A and cytokeratin 7. Ten or more cytoplasmic mucin inclusions in a tissue microarray core were seen in 51%, 48% and 31% of the 416 adenocarcinomas and 3%, 4% and 0.5% of the 194 squamous cell carcinomas with PASD, ABPAS and mucicarmine, respectively. Diagnostic pitfalls, such as entrapped benign epithelium, apoptotic/necrotic cells and glycogen, partly differed for the mucin stains. TTF-1 and napsin A IHC stainings had similar specificity but better sensitivity for adenocarcinoma than the mucin stains, but addition of PASD or ABPAS identified more tumors as adenocarcinomas (n = 8 and n = 10, respectively) than napsin A (n = 1) in cases with solid growth that were negative for TTF-1 and p40. We conclude that PASD and ABPAS have similar diagnostic performance and that these markers are of value in poorly differentiated cases. However, morphology and TTF-1 and p40 IHC staining is sufficient for correct diagnosis in most non-small cell lung cancers.

Histological examination of the interspinous ligament in horses with overriding spinous processes

The equine interspinous ligament (ISL) consists of an oblique crossing arrangement of collagenous bundles which are thought to counteract the tensile and rotational forces of distraction between the spinous processes (SPs) in the caudal thoracic and cranial lumbar spine. The aim of this controlled histological study was to assess the structural anatomy and innervation of the ISL in horses with clinically significant overriding (dorsal) SPs (ORSPs) and to compare the findings with the ISL of normal horses. Samples of the ISL were obtained from 10 horses that underwent subtotal ostectomy for treatment of ORSPs. Control samples were obtained from horses without spinal pathology. Histological staining of ISL sections with haematoxylin and eosin was performed to assess the morphology of the ligaments and with Alcian blue-periodic acid-Schiff to determine the proteoglycan and glycosaminoglycan content. Immunohistochemistry for S100 was performed for quantitative evaluation of nerves within the ISL.The ISL in horses with ORSPs had an altered collagen fibre alignment and arrangement of the ligamentous layers when compared to healthy controls. A significant increase in fibrocartilaginous tissue with evidence of fibrocartilaginous metaplasia was detected (P=0.001). The number of nerves in the ISL samples was significantly higher in horses with ORSPs than in controls (P=0.017). Structural alterations of the ISL, including loss of fibre alignment and fibrocartilaginous metaplasia, are associated with ORSPs in the equine thoracolumbar spine. In addition, an increase in innervation of the ISL in horses with ORSPs, compared to normal, may explain the thoracolumbar pain experienced by some horses with ORSPs.

Conjunctival myxoid stromal tumour: a distinctive clinicopathological and immunohistochemical study

Background/aimsTo describe the clinicopathological and immunohistochemical characteristics of 10 patients representing a new entity of benign conjunctival myxoid stromal tumours.MethodsRetrospective review of clinical findings, histopathological and immunohistochemical studies identified 10...

Sa1254 INFLUENCE OF AN UPFRONT ALCIAN-BLUE PERIODIC ACID SCHIFF STAIN ON THE HISTOPATHOLOGIC DIAGNOSIS OF SELECTED UPPER GASTROINTESTINAL DISORDERS

The Alcian Blue-periodic acid Schiff (ABPAS) stain at pH 2.5 is widely used in diagnostic histopathology. However, very few studies have evaluated how the use of this stain affects diagnoses in the practice of pathology. Our goal was to quantify the impact of upfront ABPAS staining on the diagnosis of selected upper gastrointestinal conditions. The study was conducted at the Gastrointestinal Division of Miraca Life Sciences, a specialized referral pathology laboratory that receives biopsy specimens from private outpatient endoscopy centers throughout the United States, and included the evaluation of three groups of patients, each of whom had upper gastrointestinal biopsies obtained during a diagnostic esophagogastroduodenoscopy (EGD). The biopsy specimens from esophagus, stomach, and duodenum underwent one of three processing and staining protocols: In group 1, specimens were internally processed and stained with hematoxylin and eosin (H&E) only; in group 2 they were processed externally (i.e., in another laboratory) and stained with H&E only; and specimens from group 3 were prepared internally and both were stained with H&E and ABPAS. Differences in the prevalence of Candida esophagitis, Barrett’s esophagus (BE), gastric intestinal metaplasia (GIM), and duodenal foveolar metaplasia were compared by unadjusted odds ratios (OR) and their 95% confidence intervals (95% CI). The study population comprised more than one million patients with esophageal, gastric, or duodenal biopsies. Candida spp. was detected more commonly in specimens stained with upfront ABPAS (2.03%) than in all those without (1.78%; OR 1.14, 95%CI 1.10-1.18). GIM was diagnosed in 10.6% of gastric specimens stained with ABPAS, but only in 8.06% of those without (OR 1.36, 95%CI 1.34-1.37). BE prevalence was lower in biopsies stained with ABPAS (12.3%) than in those with H&E only (14.3%). Duodenal foveolar metaplasia was detected in 14.4% of ABPAS-stained duodenal biopsies and in 8.5% of the H&E-stained specimens (OR 1.68, 95%CI 1.65-1.71). Control disease entities (Herpes esophagitis, Helicobacter pylori gastritis, and celiac disease) showed no significant differences between the three groups, suggesting that there is not a high level of selection bias. Upfront ABPAS stain in biopsy specimens from the upper gastrointestinal tract has a significant influence on the frequency of histopathologic diagnoses that rely on features highlighted by such a stain. While increased diagnostic yield of candidiasis and GIM using ABPAS is perhaps expected, the lower rate of BE suggests that pseudo-goblet cells may be over-interpreted when H&E is used alone.

Effects of glycyrrhizic acid on ERK1/2 and p38 MAPK signaling pathway in a murine model of asthma

Objective: To explore the effects of glycyrrhizic acid (GA) on Extracellular regulated protein kinases (ERK1/2) and p38 mitogen-activated protein kinases (p38 MAPK) signaling pathways in a murine model of asthma. Methods: Sixty female BALB/c mice were randomly divided into 6 groups (n=10 each): a control group, an asthmatic group, two treatment groups with low and high doses of GA, U0126 group and SB203580 group. Within 24 hours after the last OVA challenge, histological studies of lung were conducted with the hematoxylin and eosin staining (HE) and alcian blue-periodic acid-Schiff (AB-PAS), the relative protein expression of ERK1/2 and p38 MAPK were detected by immunohistochemistry and Western blotting in vivo. CD4(+) T cells were purified from spleens of OVA-sensitized and challenged mice by using the Mouse CD4 Cell Positive Isolation Kit and incubated with anti-CD3 mAb (1 μg/ml) in the presence of various concentrations of GA (10 and 100 μg/ml), U0126 (10 μmol/L) or SB203580(10 μmol/L). After 72 h of incubation, the relative protein expression of ERK1/2 and p38 MAPK of CD4(+) T cells were detected by Western blotting in vitro. Results: The asthmatic mice induced infiltration of inflammatory cells around airways and blood vessels, airway goblet cell hyperplasia and mucus production. Administration of GA at a dose of 100 mg/kg, U0126 or SB203580 significantly reduced the infiltration of inflammatory cells in the peribronchial areas and goblet cell hyperplasia compared with the asthmatic mice. The protein expressions of p-ERK1/2 were lower in GA at a dose of 100 mg/kg (0.090±0.022) and U0126 group (0.072±0.017) than those in asthmatic group (0.143±0.022) (all P<0.05). The protein expressions of p-p38 MAPK were lower in GA at a dose of 100 mg/kg (0.072±0.019) and SB203580 group (0.061±0.015) than those in asthmatic group (0.121±0.022) (all P<0.05) by immunohistochemistry. Compared with asthmatic group (0.783±0.133, 0.649±0.095), the protein expressions of p-ERK1/2 and p-p38 MAPK in GA at a high dose group (0.385±0.186, 0.275±0.089) and in U0126 group (0.117±0.051) or in SB203580 group (0.108±0.043) were decreased by Western blotting (all P<0.05). The expressions of p-ERK1/2 in CD4(+) T cells after 72 h incubation were lower in 100 μg/ml concentrations of GA (0.579±0.184) and group U0126 (0.249±0.082) and the expressions of p-p38 MAPK were much lower in 100 μg/ml concentrations of GA (0.445±0.081) and group SB203580 (0.249±0.082) compared with those in group CD3 (1.028±0.147, 0.902±0.107) (all P<0.05). Conclusion: ERK1/2 and p38 MAPK signaling pathways are activated in asthmatic mice and GA may negative regulate this activation.

Morphological and functional aspects of the epidermis of the sea lamprey Petromyzon marinus throughout development.

The development of the epidermis of sea lamprey Petromyzon marinus along the whole life cycle was studied using conventional staining techniques and lectin histochemistry. The epidermis undergoes variations in morphology and thickness throughout development. The simple cuboidal epithelium found in the epidermis of prolarvae becomes stratified cubic in the adult by increasing the number of cell layers. The cuticle thickness undergoes a steady increase during the larval period. There are changes in the glycoconjugate composition of the three main cell types of the P. marinus epidermis, mucous, granular and skein cells, which are more pronounced after metamorphosis. The Alcian blue-periodic acid Schiff (AB-PAS) histochemical method shows the presence of both acidic and neutral glycoconjugates in the mucous cells, indicating their secretory function. Moreover, lectin analysis reveals a mucous secretion containing glycoconjugates such as sulphated glycosaminoglycans (N-acetylglucosamine and N-acetylgalactosamine) and N-glycoproteins rich in mannose. Although granular cells are AB-PAS negative, they exhibit a similar glycoconjugate composition to the mucous cells. Moreover, granular cells show sialic acid positivity in larvae but this monosaccharide residue is not detected after metamorphosis. The skein cells, a unique cell of lampreys, are negative to AB-PAS staining but they mostly contain l-fucose and sialic acid residues, which also disappear after metamorphosis. The function of the granular and skein cells is still unknown but the role of their glycoconjugate composition is discussed. In addition, a different cellular origin is suggested for these two types of cells.

Establishment and effect observation on three kinds of Ti-Xu with constipation rat models

To establish three types constipation models with the syndrome of Xue-Xu, Yin-Xu and Yang-Xu in rats, and to compare the difference of colonic motility, colonic water metabolism, colonic mucus secretion and the level of aquaporin-2 (AQP2). Forty Sprague Dawley rats, male and female in half, were randomly divided into four groups:normal control group(N), Xue-Xu with constipation group(XC), Yin-Xu with constipation group(YC) and Yang-Xu with constipation group(YAC). Bloodletting and loperamide were used to induce the Xue-Xu with constipation in rats, thyroxin and loperamide were used to induce the Yin-Xu with constipation in rats, ice-water stimulation and loperamide were used to induce the Yang-Xu with constipation in rats. Bloodletting was coducted once a week,drug was ad-ministrated orally to rats once a day for continuous 42 days. Rats'state, body weights, stool character, transfer time between mouth and anal, small intestinal propulsion rate were measured. The colon tissues of rats were stained with alcian blue-periodic acid schiff (AB-PAS) to analyze the changes of mucus secretion. The aquaporin-2 expressions in proximal and distal colon of rats were measured by immunohistochemical method. Compared to normal control group, weight-increasing speed of the rats were reduced in three models. The order of indepen-dent movement change was YC, XC and YAC. Solid stool appeared at 30 d, fecal scores were increased. Transfer time between mouth and anal was significantly extended, small intestinal propulsion rate were significantly decreased(P < 0.05, P < 0.01). Water content in stool were significantly decreased in three type models(P < 0.05, P < 0.01), the content of water in colon were decreased in XC and YAC group (P < 0.05, P < 0.01). The colonic gland and goblet cells were narrowed in some extent, the mucus excretion were decreased. The levels of aquaporin-2 expressions in proximal and distal colon of rats were increased(P < 0.05, P < 0.01), increasing order of AQP2 expression in proximal colon was YAC, YC and XC, increasing order of AQP2 expression in distal colon was YAC, XC and YC. Long term stimulation of complex factors could induce Ti-Xu with constipation animal model, and there were some differences in colonic motility, water metabolism, colonic mucus secretion and AQP2 in rats.

Histological observation of a gelatin sponge transplant loaded with bone marrow-derived mesenchymal stem cells combined with platelet-rich plasma in repairing an annulus defect.

ObjectiveTo research the histological characteristics of a gelatin sponge transplant loaded with goat BMSCs (bone marrow-derived mesenchymal stem cells) combined with PRP (platelet-rich plasma) in repairing an annulus defect.MethodBMSCs were separated from the iliac crest of goats, sub-cultured and identified after the third generation. Then, PRP was obtained using blood from the jugular vein of goats via two degrees of centrifugation. In the animal experiments, the goats were divided into the following three groups: a sham group, an injury group and a therapeutic group. In the sham group, we decompressed the lamina and exposed the annulus fibrosus. In the injury group, we exposed the annulus fibrosus after decompression of the lamina and created a 1 × 1 cm defect in the annulus using surgical instruments. In the therapeutic group, after decompression of the lamina, we exposed the annulus, created a 1 × 1 cm defect using surgical instruments, and placed a gelatin sponge combined with BMSCs and PRP into the defect for a combined method of repair. Three, six and twelve weeks after the surgery, the previously damaged or undamaged annulus tissue was removed from the three groups. Then, the above tissue was assayed using HE (hematoxylin-eosin) staining, Masson trichrome staining, AB-PAS (Alcian blue-periodic acid Schiff) staining, and type II collagen staining and observed by microscopy.ResultsFrom the HE staining, we observed that the number of repair cells gradually increased. Compared to the injury group, the cell density and gross morphology of cells in the therapeutic group were closer to those of the sham group. As observed by Masson trichrome gelatin staining, many of the fibroblast cells or tissues were under repair, and as time progressed, the number of fibroblast cells and amount of tissue gradually increased. The results of the AB-PAS staining suggest that chondrocytes participated in the repair of the annulus. The level of type II collagen gradually increased, as determined by immunohistochemical staining.ConclusionOur results demonstrate that a gelatin sponge transplant loaded with BMSCs and PRP can effectively repair annulus defects.

The distribution of mucous secreting cells in the gastrointestinal tracts of three small rodents from Saudi Arabia: Acomys dimidiatus, Meriones rex and Meriones libycus

The proportion of mucin phenotypes (which form the protective biofilm of the gastrointestinal tract) differs between intestinal regions. This study examines the distribution of mucin secreting cells in the gastrointestinal tracts of the Eastern spiny mouse (Acomys dimidiatus), King jird (Meriones rex) and Libyan jird (Meriones libycus), which inhabit the dry and hot deserts of Saudi Arabia. Intestinal tract samples were processed to wax and tissue sections stained with Alcian Blue-Periodic Acid Schiff (AB-PAS) and High Iron Diamine-Alcian Blue (HID-AB) in order to determine different mucin phenotypes by quantitative analysis. Mixed mucin secreting cells (combined neutral and acid) was the predominant mucin secreting cell type observed throughout the gastrointestinal tract in all species. Acid mucin secreting goblet cells were mainly located in the colon. A. dimidiatus presented with significantly more total sialo than sulfomucin secreting cells while the opposite was true for both Meriones species. The distribution of mucin secreting cells is therefore similar to previously reported results for small mammals not living under arid conditions.

Histological, surface ultrastructural, and histochemical study of the stomach of red piranha, Pygocentrus nattereri (Kner)

Abstract The structural characterization and function of the stomach in the omnivore Pygocentrus nattereri were described using light and scanning electron microscopy. The sac-like stomach was morphologically divided into the cardiac and pyloric regions. The histological structure of the stomach consisted of four layers of the mucosa, submucosa, muscularis, and serosa. The superficial epithelium of the cardiac stomach was lined with columnar epithelial cells and the glandular epithelium contained numerous gastric glands. Gastric glands were completely absent in the pyloric portion. The mucosal surface of the stomach was a meshwork of various folds, provided with oval or rounded columnar epithelial cells which were densely packed with short, stubby microvilli. The occasional presence of conspicuous gastric pits was surrounded by epithelial cells. The localization and chemical nature of acid and neutral mucins in the various cells of the stomach was studied by employing combined the Alcian Blue-Periodic Acid Schiff (AB-PAS) technique. The deposition of glycogen was detected in the gastric glands as well as in the epithelial lining of the stomach. The utmost reactions for protein and tryptophan were recorded in the gastric glands of mucosa. The cellular organization and histochemical characterization of the stomach are discussed in relation to the feeding and digestion of the fish concerned.

Congenital Ichthyosis in 14 Great Dane Puppies With a New Presentation.

The present study describes a generalized congenital skin condition in 14 Great Dane puppies. Macroscopically, all dogs showed generalized gray to yellow scaling and skin wrinkles on the head and all 4 extremities. Skin sections were histologically examined using hematoxylin and eosin, Heidenhain's Azan, and Sudan red III staining methods and by conducting the alcian blue/periodic acid Schiff (AB/PAS) reaction technique on sections. Furthermore, incubation with hyaluronidase was performed. Skin samples were ultrastructurally analyzed using transmission electron microscopy. All affected Great Dane puppies had epidermal and follicular orthokeratotic hyperkeratosis, enlarged keratohyaline granules, vacuolated keratinocytes, and accumulations of an eosinophilic and alcianophilic, lipid-rich material within dilated hair follicular lumina and the cytoplasm of sebocytes. The macroscopic, histopathologic, and ultrastructural skin changes in all 14 Great Dane puppies indicate a new variant of a primary disorder of cornification with congenital, non-epidermolytic, lamellar ichthyosiform appearance.

Hyperbaric oxygen against stress ulcer in blast-related traumatic brain injury

Objective To investigate the effect of hyperbaric oxygen to reduce stress-induced gastric mucosal injury in rabbits with blast-related brain injury.Methods Posttraumatic stress ulcer in a rabbit model of blast-related brain injury was created using paper detonators.Ninety New Zealand white rabbits were randomly divided into normal control group (n =10),blast-related injury group (n =40) and hyperbaric oxygen treatment group (hyperbaric oxygen group,n =40) according to the random number table.Rabbits in blast-related injury group and hyperbaric oxygen group were subgrouped at 6 hours,24 hours,72 hours and 7 days postinjury,with 10 rabbits at each time point.Gastric mucosal injury was detected in aspects of ulcer index,TUNEL method for apoptotic index,and alcian blue-periodic acid-schiff (AB/PAS) staining and semi-quantitative analysis of pathological histology using.Results Neither stress ulcer nor apoptosis occurred in normal control group.At 6 hours,24 hours,72 hours and 7 days postinjury,index of gastric ulcer in blast-related injury group was 12.08 ± 1.02,25.56 ± 0.88,20.36 ±0.96 and 17.56 ± 1.12 respectively,far higher than 8.02 ±0.58,15.22 ± 1.05,12.39 ±0.65 and 9.22 ± 0.79 in hyperbaric oxygen group (P ＜ 0.05) ; apoptotic index in blast-related injury group was 20.22 ± 1.56,27.56 ± 0.96,24.36 ± 1.23 and 16.38 ± 1.24,far higher than 10.78 ± 0.93,13.89 ±0.84,10.55 ± 1.58 and 8.56 ± 1.47 in hyperbaric oxygen group (P ＜ 0.05).Thickness of AB/PAS-positive mucin layers showed marked attenuation even loss in blast-related injury group,but the loss was minor in hyperbaric oxygen group (P ＜ 0.05).Conclusion Hyperbaric oxygen is effective to decrease the stress ulcer index and apoptosis index of gastric mucosa,increase the thickness of AB/PAS positive layer and lighten the gastric mucosal injury of rabbits with blast-related brain injury. Key words: Craniocerebral trauma; Ulcer; Blast injuries ; Hyperbaric oxygenation

Abstract 863: Clinical significance of mucinous production in rectal cancer after preoperative chemoradiotherapy

Abstract Background: Mucinous adenocarcinoma (MA) is a relatively rare histological subtype of colorectal cancer, in which more than 50% of the tumor is occupied by extracellular mucin. MA has been reported to be associated with poor response to neoadjuvant chemoradiotherapy (CRT), but the prognostic significance still remains to be evaluated. Here, we aimed to analyze the prognostic significance of mucin production in the response of rectal cancer to neoadjuvant CRT. Methods: The medical records of 141 consecutive patients receiving rectal resection after neoadjuvant CRT for the treatment of rectal cancer at our institution, in the period between October 2003 and December 2012, were retrospectively reviewed. For the histological evaluation, Hematoxylin-Eosin (HE) and Periodic Acid Schiff/Alcian Blue (PAS/AB) stainings were used. Mucin production was evaluated according to the proportion of the tumor area occupied by extracellular mucin, and cases were sorted, according to the mucin occupancy rate, into three groups, namely, low (0 - 5%), medium (5 - 25%) and high (&amp;gt;25%). Results: The number of patients in the low, medium and high mucin rate groups was 89 (63.1%), 28 (19.9%) and 24 (17.0%), respectively. Only advanced tumor stage out of histological features had a significant correlation with the high mucin rate group. Compared to the low mucin rate group, patients in the high group had a worse prognosis in disease free survival, and also a worse prognosis in distant recurrence free survival, compared to the medium group, while no significant difference was observed among the three groups in overall and local recurrence-free survivals. Multivariate analysis showed that the high mucin rate group had an independent association with shorter disease-free survival, and those advanced stage tumors (T3 and T4) with a high mucin rate had an independent association with shorter distant recurrence-free survival. Conclusion: Mucinous production in rectal cancer may be associated with resistance to neoadjuvant CRT. Thus, careful histological analysis for the presence of extracellular mucin should be indicated for rectal patients receiving neoadjuvant CRT, and those cases with high mucin rate (&amp;gt; 25% mucinous component) should be closely followed-up. Citation Format: Kensuke Kaneko. Clinical significance of mucinous production in rectal cancer after preoperative chemoradiotherapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 863. doi:10.1158/1538-7445.AM2014-863

Benign multicystic peritoneal mesothelioma associated with partial spontaneous regression in an aged cow

This article describes a benign multicystic peritoneal mesothelioma presenting partial spontaneous regression in a 16-year-old Japanese Shorthorn cow, which exhibited innumerable fluid-filled cysts (<20 cm diameter) distributing throughout the serosal surface of the peritoneal cavity on postmortem examination. Each cyst was further subdivided into several daughter cysts. On histopathology, these cystic structures consisted of multiloculated irregular spaces that contained serous fluid positive for Alcian blue stain and periodic acid-Schiff (PAS) reaction and were lined by neoplastic mesothelial cells arranged in a single to multicell layers. In some places, neoplastic cells exhibited a tubular, papillary, villous-like, or peninsular growth pattern. Neoplastic cells exhibited mild anisocytosis and anisokaryosis with very low mitotic rate. Immunohistochemically, all or some of neoplastic cells were positive for cytokeratin or vimentin, respectively. Some smaller cysts represented abundant sclerotic stroma in association with fewer shrunken neoplastic cell-lined tubules enclosed by dense fibrocollagenous stroma, collagen fiber collapse, infiltrates of lymphocytes and macrophages disposing of collagen fiber debris, and many capillaries. These findings raised the likelihood that a specific process operating partial spontaneous regression might have been implicated in this tumor. The precise mechanism underlying this phenomenon remains uncertain, although the role of abundant sclerotic stroma was highlighted.

Cutaneous mucinosis in a strain of brown-egg laying chickens.

Cutaneous mucinosis is a cutaneous disorder described in humans, dogs, and rarely cats but never reported in birds. Twenty-six brown egg-laying chickens between ages 43 and 46 weeks had a history of feather loss, scaly, dry skin, weight loss, and decreased egg production. Microscopic findings in the skin included fragmentation of collagen bundles and interstitial, periadnexal, and perivascular dermal accumulation of wispy, mildly basophilic material that was also occasionally observed within the follicular epithelium. A moderate lymphoplasmacytic and heterophilic perivascular dermatitis was also observed. The wispy to granular material was diffusely Alcian blue positive and periodic acid-Schiff negative (consistent with mucin), suggesting a diagnosis of primary or secondary cutaneous mucinosis. The cause of this condition could not be determined.

Clinicopathological and EBV analysis of respiratory epithelial adenomatoid hamartoma

BackgroundTo investigate the clinicopathological characteristics of respiratory epithelial adenomatoid hamartoma (REAH) in residents of Southern China and to study the correlation between REAH and Epstein-Barr virus (EBV).MethodsClinicopathological data of 53 cases of REAH were retrospectively analyzed. The immunoreactivity for CK 7, CK20, CEA, p53, and Ki-67, Alcian blue–periodic acid-Schiff (AB-PAS) staining and in situ hybridization for EBV-encoded RNA (EBER) were carried out.ResultsREAH lesions were covered with ciliated columnar epithelium and proliferation of subepithelial glands, which were positive for CK7, and negative for CK20, CEA, and p53. Goblet cell metaplasia was stained blue by AB-PAS. The frequency of EBER positive cases in REAH located in nasopharynx was 27.78%, compared with that in the nasal cavity (15.79%) and paranasal sinuses (12.50%), there were no statistical differences.ConclusionsREAH is an uncommon entity with distinctive morphologic features and EBV may have nothing to do with REAH.Virtual slidesThe virtual slide(s) for this article can be found here:http://www.diagnosticpathology.diagnomx.eu/vs/5875687401178748

Oxidative DNA Damage as a Potential Early Biomarker of Helicobacter pylori Associated Carcinogenesis

Helicobacter pylori infection is an established risk factor for gastritis, gastric ulcer, peptic ulcer and gastric cancer. CagA +ve H. pylori has been associated with oxidative DNA damage of gastric mucosa but their combined role in the development of gastric cancer is still unknown. Here we compare the combined expression of cagA and 8-hydroxy-2'-deoxyguanosine (8-OHdG) in normal, gastritis and gastric cancer tissues. Two hundred gastric biopsies from patients with dyspeptic symptoms, 70 gastric cancer tissue samples and 30 gastric biopsies from non-dyspeptic individuals (controls) were included in this study and 8-OHdG was detected by immunohistochemistry (IHC). Histological features and the presence of H. pylori infection were demonstrated by Hematoxylin and Eosin (HE), Giemsa and alcian blue-periodic acid-Schiff ± diastase (AB-PAS ± D) staining. DNA was extracted from tissues and polymerase chain reaction (PCR) performed to determine the presence of ureaseA and cagA genes of H. pylori. The results showed the presence of H. pylori in 106 (53 %) gastric biopsies out of 200 dyspeptic patients, including 70 (66 %) cases of cagA + ve H. pylori. The presence of cagA gene and high expression of 8-OHdG was highly correlated with severe gastric inflammation and gastric cancer particularly, in cases with infiltration of chronic inflammatory cells (36.8 % cagA + ve, 18 %), neutrophilic activity (47.2 %, 25.5 %), intestinal metaplasia (77.7 %, 35.7 %) and intestinal type gastric cancer (95 %, 95.4 %) (p ≤ 0.01). In conclusion, H. Pylori cagA gene expression and the detection of 8-OHdG adducts in gastric epithelium can serve as potential early biomarkers of H. Pylori-associated gastric carcinogenesis.

Aloe vera affects changes induced in pulmonary tissue of mice caused by cigarette smoke inhalation

This study was undertaken to determine the influence of Aloe vera (AV) on changes induced in pulmonary tissue of cigarette smoke (CS) inhaling mice. CS inhalation for 4 weeks caused pulmonary damage as evident by histoarchitectural alterations and enhanced serum and tissue lactate dehydrogenase (LDH) activities. CS inhalation also led to increased mucin production as revealed by mucicarmine and Alcian Blue-Periodic Acid Schiff (AB-PAS) staining. Studies on bronchoalveolar lavage fluid (balf) of CS exposed animals revealed structural changes in phospholipids and increase in surface tension when compared with control counterparts. These changes were accompanied by enhanced nitric oxide (NO) levels, citrulline levels, peroxidative damage, and differential modulation of antioxidant defense system. AV administration (seven weeks, 500 mg/kg b.w. daily) to CS inhaling mice led to modulation of CS induced pulmonary changes as revealed by lesser degree of histoarchitectural alterations, lesser mucin production, decreased NO levels, citrulline levels, peroxidative damage, and serum LDH activity. AV treatment to CS inhaling mice was associated with varying response to antioxidant defense system, however balf of CS + AV treated animals did not exhibit appreciable changes when compared with that of CS exposed animals. These observations suggest that AV has the potential to modulate CS induced changes in the pulmonary tissue which could have implications in management of CS associated pulmonary diseases, however, further investigations are required to explore its complete mechanism of action.