Periodic acid-Schiff Diastase Research Articles

Erythrasma of the Vulva: An Invisible Dermatosis.

Erythrasma is a prevalent superficial bacterial infection typically caused by Corynebacteria species and preferentially affecting intertriginous sites including axillary, interdigital, and inguinal skin folds. However, erythrasma of the vulva is uncommon, with only 2 cases previously reported. Although erythrasma can be diagnosed clinically using Woods lamp examination, it may not always be considered in the differential diagnosis for patients presenting with persistent vulvar pruritus. We report 12 cases of vulvar erythrasma identified by histopathology, with a review of clinical and histologic features. The mean patient age was 60.1yr and the mean patient BMI was 30.5. Five of 12 patients presented with pruritic rash. The time from symptom onset to diagnosis was 9mo in 1 case, >18mo in 4 cases, and unknown in the remaining cases. The characteristic histologic features were compact orthokeratosis and mild perivascular chronic inflammation. In all 12 cases, Periodic Acid-Schiff-diastase (PAS-D) staining highlighted intracorneal filamentous rods which were not readily appreciable on H&E. After the diagnosis of erythrasma, 4 patients were treated with topical lincomycin, of whom 3 had clinical improvement in symptoms. One patient was treated with topical macrolide antibiotic and also reported improvement in symptoms. Consideration of erythrasma on the differential for patients presenting with vulvar rash and pruritus may shorten the time to diagnosis and treatment, minimize patient discomfort, and reduce the scope and cost of diagnostic testing.

Placental scaffolds as a potential biological platform for embryonic stem cells differentiation into hepatic-like cells lineage: A pilot study

Hepatic microenvironment plays an essential role in liver regeneration, providing the necessary conditions for cell proliferation, differentiation and tissue rearrangement. One of the key factors for hepatic tissue reconstruction is the extracellular matrix (ECM), which through collagenous and non-collagenous proteins provide a three-dimensional structure that confers support for cell adhesion and assists on their survival and maintenance. In this scenario, placental ECM may be eligible for hepatic tissue reconstruction, once these scaffolds hold the major components required for cell support. Therefore, this preliminary study aimed to access the possibility of mouse embryonic stem cells differentiation into hepatocyte-like cells on placental scaffolds in a three-dimensional dynamic system using a Rotary Cell Culture System. Following a four-phase differentiation protocol that simulates liver embryonic development events, the preliminary results showed that a significant quantity of cells adhered and interacted with the scaffold through outer and inner surfaces. Positive immunolabelling for alpha fetus protein and CK7 suggest presence of hepatoblast phenotype cells, and CK18 and Albumin positive immunolabelling suggest the presence of hepatocyte-like phenotype cells, demonstrating the presence of a heterogeneous population into the recellularized scaffolds. Periodic Acid Schiff-Diastase staining confirmed the presence of glycogen storage, indicating that differentiate cells acquired a hepatic-like phenotype. In conclusion, these preliminary results suggested that mouse placental scaffolds might be used as a biological platform for stem cells differentiation into hepatic-like cells and their establishment, which may be a promissing biomaterial for hepatic tissue reconstruction

Retinal Phenotyping of a Murine Model of Lafora Disease.

Lafora disease (LD) is a progressive neurologic disorder caused by biallelic pathogenic variants in EPM2A or EPM2B, leading to tissue accumulation of polyglucosan aggregates termed Lafora bodies (LBs). This study aimed to characterize the retinal phenotype in Epm2a-/- mice by examining knockout (KO; Epm2a-/-) and control (WT) littermates at two time points (10 and 14 months, respectively). In vivo exams included electroretinogram (ERG) testing, optical coherence tomography (OCT) and retinal photography. Ex vivo retinal testing included Periodic acid Schiff Diastase (PASD) staining, followed by imaging to assess and quantify LB deposition. There was no significant difference in any dark-adapted or light-adapted ERG parameters between KO and WT mice. The total retinal thickness was comparable between the groups and the retinal appearance was normal in both groups. On PASD staining, LBs were observed in KO mice within the inner and outer plexiform layers and in the inner nuclear layer. The average number of LBs within the inner plexiform layer in KO mice were 1743 ± 533 and 2615 ± 915 per mm2, at 10 and 14 months, respectively. This is the first study to characterize the retinal phenotype in an Epm2a-/- mouse model, demonstrating significant LB deposition in the bipolar cell nuclear layer and its synapses. This finding may be used to monitor the efficacy of experimental treatments in mouse models.

Deep Dermatophytosis Presented as Multiple Exophytic Masses caused by Trichophyton rubrum in Immunocompromised Patient with Rheumatoid Arthritis; A Case Report

Dermatophytes invade the stratum corneum and infect the skin, nails, and hair, mostly resulting in superficial infections. Deep dermatophytosis involving the dermis and subcutaneous layer has rarely been reported in immunocompromised individuals. Herein, we report a case of deep dermatophytosis caused by Trichophyton rubrum. A 71-year-old woman presented with multiple erythematous exophytic and subcutaneous nodules in both the lower legs. The patient was taking immunosuppressive agents for rheumatoid arthritis and antifungal agents for tinea pedis and onychomycosis, which was improperly ceased. Histopathological findings showed diffuse granulomatous infiltration with multinucleated cells, lymphocytes, histiocytes, and neutrophils in the dermis. Septate and branched hyphae were observed in the dermis using periodic acid-Schiff diastase and Gomori methenamine silver staining. T. rubrum was identified in fungal culture from the tissue sample and confirmed through phylogenetic analysis of the internal transcribed spacer and large subunit regions in ribosomal ribonucleic acid gene. Intravenous amphotericin B was administered for septic shock before the confirmation of the causative organism, which rapidly improved the condition.

S2772 Subcutaneous Sweet Syndrome Successfully Treated With Ustekinumab in Patient With Ulcerative Colitis

Introduction: Extraintestinal manifestations (EIMs) of inflammatory bowel disease (IBD) are well-established. Sweet syndrome (SS), or acute febrile neutrophilic dermatosis, is a reactive mucocutaneous manifestation of IBD presenting as erythematous papules/plauqes with associated fever and arthomyalgias. There is limited evidence regarding efficacy of Ustekinumab in management of EIMs of UC, particularly SS. Case Description/Methods: The patient is a 50-year-old woman with left-sided UC (Montreal classification E2) that previously failed several therapies, including mesalamine, azathioprine, infliximab, adalimumab, vedolizumab, and a fecal microbiota transplant trial. She presented with worsening abdominal pain, diarrhea/hematochezia, and bilateral lower extremity pain. She was febrile and tachycardic with tender, violaceous, and ulcerated subcutaneous nodules on the bilateral ankles with hemorrhagic bullae (FigureA). Labs revealed negative C. diff, elevated ESR/CRP, thrombocytosis, and borderline leukocytosis with neutrophilic predominance. Lower extremity MR imaging and bone biopsy revealed multifocal sterile osteomyelitis with adjacent abscess; skin biopsy demonstrated dense granulomatous/neutrophilic infiltrate in the deep dermis/subcutis without microorganisms, consistent with subcutaneous SS (FigureB). A restaging colonoscopy demonstrated severe proctosigmoid ulcerative colitis (FigureC). Given that the patient had failed several immunomodulators and biologic therapies, she was started on prednisone and Ustekinumab. Four months later, she reported complete resolution of her cutaneous SS off steroids and improving UC symptoms (less frequent and more formed stools, minimal hematochezia). Repeat MRI demonstrated resolution of the sterile osteomyelitis and abscess. Discussion: Our case highlights the novel use of Ustekinumab in the treatment of subcutaneous SS with sterile osteomyelitis in a patient with flaring UC. SS is a neutrophilic dermatosis that is a rare EIM of IBD, classically characterized by tender cutaneous lesions. There is limited evidence supporting the use of Ustekinumab in treating EIMs of CD, and a lack of data regarding its efficacy in treating EIMs of UC. Our case fills a major gap in the literature and highlights the clinical efficacy of Ustekinumab for SS and UC especially in patients that have a contraindication to, failed previous treatment with, or otherwise cannot tolerate corticosteroids and TNF-alpha antagonists.Figure 1.: 1A. Right lateral malleolus with 2.5 cm shallow erosion with slightly violaceous borders and central bright yellow fibrinous debris. There was slight atrophy of surrounding skin with mild edema and hyperpigmentation (left image). Left medial malleolus/dorsal foot with grouped irregular shallow ulcers with surrounding mild violaceous erythema (middle and right image). 1B. Left ankle biopsies showed mixed septal and lobular panniculitis with granulomatous and neutrophilic inflammation. Histologic sections show a mildly spongiotic epidermis with dense granulomatous and neutrophilic inflammatory infiltrate in the deep dermis and subcutis (left image). The inflammatory infiltrate, which consists of prominent neutrophils, histiocytes, and scattered lymphocytes, surrounds vascular and adnexal structures, as well as adipocyte lobules. Definitive features of vasculitis are not seen. PASd (periodic acid-Schiff-diastase), GMS (Grocott’s methenamine silver), Gram, and Fite staining fail to highlight microorganisms (right image). 1C. Colonoscopy demonstrated severe (Mayo Endoscopy Score 3) proctosigmoid ulcerative colitis to 30 cm (left image). Sigmoid biopsies showed significant immune cell infiltration with crypt architectural distortion concerning for severe chronic active colitis. No evidence of granulomas, dysplasia or cytomegalovirus (right image).

27437 Disseminated Candida tropicalis in an immunocompromised pediatric patient

We report a case of a 12-year-old female with newly diagnosed B-cell acute lymphoblastic leukemia status post induction chemotherapy with hospital course complicated by Candida tropicalis fungemia requiring fluconazole. Two weeks after treatment initiation, she presented with a new onset fever and discrete erythematous papules, some with pseudovesicular appearance, along with erythematous macules and thin plaques localized to the upper extremities. At presentation, labs demonstrated pancytopenia, elevated beta-D-glucan assay (>500), and Candida tropicalis growth on both wound and tissue culture. Blood cultures were negative. Computed tomography with contrast demonstrated hypodense areas involving the liver, spleen, and pectoralis major muscle concerning for fungemic microabscesses. A right arm punch biopsy revealed a dense dermal inflammatory infiltrate consisting of numerous neutrophils, histiocytes, lymphocytes and eosinophils. Periodic acid Schiff diastase and Gomori methenamine silver stains highlighted many associated fungal organisms. A diagnosis of disseminated candida tropicalis was rendered. Treatment was changed from oral fluconazole to intravenous micafungin. Although her cutaneous involvement improved, she remained febrile. She was switched to amphotericin B and prednisone with marked improvement. This case highlights the difficulty in management of disseminated Candida tropicalis in immunosuppressed pediatric patients. The observed disseminated disease may be explained by emerging resistance of Candida tropicalis to fluconazole. Improvement of fever with prednisone supports the hypothesis that disseminated candidiasis causes an inflammatory response and emphasizes the importance of adjuvant corticosteroid therapy. Cutaneous involvement may indicate underlying disseminated disease, and dermatologists are crucial in the diagnosis and treatment of this opportunistic, often fatal disease.

Kupffer Cell Hyaline Globules in Children With Autoimmune Hepatitis

Hyaline globules (HGs) in the cytoplasm of Kupffer cells (KCs) have been appraised for being a typical feature of autoimmune hepatitis (AIH). This study aimed to determine how useful Kupffer cell hyaline globules (KCHGs) are in diagnosing AIH vs. other causes of pediatric chronic liver diseases (PCLDs). This retrospective study recruited 124 children; 58 with AIH, 50 with chronic hepatitis C virus (HCV) infection, and 16 with Wilson's disease (WD). Two pathologists retrieved paraffin blocks of liver biopsies and prepared new cut sections for Periodic acid-Schiff-Diastase (PAS-D) stain. They independently examined liver biopsies before starting treatment. Two pediatricians reviewed medical records for demographic, clinical, laboratory, and serological findings. Females represented 48.6% of the studied children with a median age of 5.8 (4.9) years. Pathologists identified KCHGs in 67.24%, 12.5%, and 6.0% of AIH, WD, and HCV affected children respectively, P<0.001. A significantly higher proportion of seropositive than seronegative AIH patients had KCHGs (77.5% vs. 50.0%), (P<0.05). In multivariate analysis, KCHGs and prolonged prothrombin time were the only significant predictors that differentiate between AIH and the other studied PCLDs. The odds ratio of having AIH increased 68 times if KCHGs were seen. Among children with AIH, the presence of KCHGs was associated with higher median levels of direct bilirubin 2.2 (1.3) vs. 1.2 (2.2), and immunoglobulin G 3.2 (1.9) vs. 2.0 (1.7), (P<0.05), but not to histopathological findings or hepatic fibrosis and activity. KCHGs are key indicators that can differentiate between AIH and other PCLDs, and between seropositive and seronegative AIH.

An inducible glycogen synthase-1 knockout halts but does not reverse Lafora disease progression in mice

Malstructured glycogen accumulates over time in Lafora disease (LD) and precipitates into Lafora bodies (LBs), leading to neurodegeneration and intractable fatal epilepsy. Constitutive reduction of glycogen synthase-1 (GYS1) activity prevents murine LD, but the effect of GYS1 reduction later in disease course is unknown. Our goal was to knock out Gys1 in laforin (Epm2a)-deficient LD mice after disease onset to determine whether LD can be halted in midcourse, or even reversed. We generated Epm2a-deficient LD mice with tamoxifen-inducible Cre-mediated Gys1 knockout. Tamoxifen was administered at 4 months and disease progression assessed at 12 months. We verified successful knockout at mRNA and protein levels using droplet digital PCR and Western blots. Glycogen determination and periodic acid–Schiff–diastase staining were used to analyze glycogen and LB accumulation. Immunohistochemistry using astrocytic (glial fibrillary acidic protein) and microglial (ionized calcium-binding adapter molecule 1) markers was performed to investigate neuroinflammation. In the disease-relevant organ, the brain, Gys1 mRNA levels were reduced by 85% and GYS1 protein depleted. Glycogen accumulation was halted at the 4-month level, while LB formation and neuroinflammation were significantly, though incompletely, prevented. Skeletal muscle analysis confirmed that Gys1 knockout inhibits glycogen and LB accumulation. However, tamoxifen-independent Cre recombination precluded determination of disease halting or reversal in this tissue. Our study shows that Gys1 knockdown is a powerful means to prevent LD progression, but this approach did not reduce brain glycogen or LBs to levels below those at the time of intervention. These data suggest that endogenous mechanisms to clear brain LBs are absent or, possibly, compromised in laforin-deficient murine LD.

S2793 Sprue-Like Enteropathy Secondary to Lisinopril Use

INTRODUCTION: Diarrhea is reported as a rare adverse effect of angiotensin-converting enzyme (ACE) inhibitors, but there are few reports of sprue-like enteropathy from its use. In contrast, this has been well described with the angiotensin II receptor blocker (ARB), olmesartan. We describe a case of drug-induced enteropathy secondary to lisinopril. CASE DESCRIPTION/METHODS: A 76-year-old man with hypertension and coronary artery disease presented with one year of diarrhea, abdominal discomfort and a 35-pound unintentional weight loss. His first EGD yielded duodenal biopsies with villous blunting and increased IELs. Tissue transglutaminase antibody (tTg, IgA) was 22 U/mL (negative is <20 U/mL), and the patient was advised to follow a gluten free diet. He did so for 9 months without symptom improvement and so returned to a gluten-rich diet. Despite a gluten-rich diet for 4 months, the patient’s repeat tTg, IgA and deamidated gliadin peptide (DGP) IgA and IgG were negative. In addition, he was negative for HLA-DQ2 and HLA-DQ8 genotypes, making celiac disease unlikely. All immunoglobulin levels were normal. Then on repeat EGD and colonoscopy, duodenal and terminal ileal biopsies showed villous blunting and dense infiltrate of IELs. Biopsies were negative for Congo red and periodic acid-Schiff diastase stains, sent to evaluate for amyloid and Whipple's disease. Video capsule endoscopy identified diffuse small bowel villous flattening, though no erosions, ulcerations or strictures were seen. Stool testing was negative for giardia and fecal calprotectin. Glucose hydrogen breath test was negative. The patient had a normal fasting gastrin level, and his ANA and anti-enterocyte antibody were negative. We then considered lisinopril-induced enteropathy, as the patient had been on lisinopril since symptom onset. After lisinopril was stopped, the patient's symptoms resolved within 3 months. He gained back weight and no longer had abdominal discomfort. Repeat endoscopy is planned, but was postponed during the COVID-19 pandemic. DISCUSSION: It is important to maintain a high degree of suspicion for drug-induced enteropathy. One study from Italy and Germany examined patients on olmesartan, ARBs and ACE inhibitors and found rare instances of ACE inhibitors leading to intestinal malabsorption (Malfertheiner et al. Pharmacoepidemiol Drug Saf. 2018, 581-586). Olmesartan is classically the medication associated with sprue-like enteropathy, and our case highlights that lisinopril should be considered as well.

Meningeal Granular Cell Tumour in a Green Tree Python (Morelia viridis)

Granular cell tumours (GCTs) are uncommon neoplasms mostly reported in man, dogs and horses. The origin of GCT is thought to be Schwann cells, with the associated characteristics of neural crest morphology. Neoplastic cells often demonstrate positive immunoreactivity for S100, LC3, vimentin and p62. They are also periodic acid-Schiff (PAS) positive and diastase resistant. A female green tree python (Morelia viridis) was presented for severe constipation and hyporexia of 4 month's duration and, despite treatment, died the next day. A 4.8×3.4mm intracalvarial GCT was identified, compressing the overlying cerebrum without invasion. Neoplastic cells were immunoreactive to S100 and had brightly eosinophilic cytoplasmic granules that were PAS positive and diastase resistant. Electron microscopy revealed numerous cytoplasmic lysosomes in neoplastic cells. GCTs are reported rarely in non-mammalian species with three reports in birds. This represents the first report of a GCT in a reptile.

Increased Frequency of Heterozygous Alpha-1-Antitrypsin Deficiency in Liver Explants From Nonalcoholic Steatohepatitis Patients.

Cirrhotic explanted livers occasionally have unexpected periodic acid-Schiff-diastase (PASD)-positive globules within the hepatocyte cytoplasm. It is often unclear whether this finding is a nonspecific consequence of cirrhosis or is indicative of an underlying alpha-1-antitrypsin deficiency (A1ATD) contributing to the cirrhosis. In this study, explanted livers were retrospectively evaluated for histopathology (including PASD status with confirmatory alpha-1-antitrypsin [A1AT] immunohistochemistry [IHC]), and chart review provided etiology of liver failure and general clinical parameters. Real-time polymerase chain reaction was used to detect A1AT genotype (SERPINA1 S and Z alleles) by melting curve analysis on liver explant tissue from selected cases. Of 196 explanted livers, 21 (11%) had PASD+ globules, which were significantly enriched in patients with a clinical diagnosis of nonalcoholic steatohepatitis (NASH; 47%) compared with other causes (P<0.001). IHC confirmed all PASD+ globules were A1AT+, with 20 of 21 cases demonstrating diffuse A1AT staining. In an expanded NASH cohort, 42% (14/33) of explants had PASD+ globules, 92% of which were homozygous (n=1) or heterozygous (n=11) for the SERPINA1 Z allele, corresponding to nearly 40% of all NASH patients. Overall, the Z allele was present in 10% of all tested liver explants, with 85% of PASD+ cases genotyping homozygous (n=2) or heterozygous (n=20), which is far in excess of the estimated 2% in the general population. These results indicate PASD+ A1AT globules (with confirmatory genotyping showing at least 1 Z allele) are commonly observed in NASH, suggesting a synergistic relationship toward liver fibrosis. In addition, the high frequency of SERPINA1 Z alleles in liver transplantation patients supports the utility of pretransplant genotyping.

A Polyclonal Antibody to NKX3.1 Identifies Fungal Organisms From the Esophagus.

NKX3.1 is a transcription factor used to identify prostatic adenocarcinomas. We describe novel functionality for NKX3.1 compared with Grocott and periodic acid-Schiff-diastase (PASD) on esophageal biopsies. We identified esophageal biopsies on the basis of the search term "candida" from March 28, 2012 to December 27, 2013. Of 85 cases for which 3 stains were available and at least 1 stain was positive for fungus consistent with Candida, 83 cases stained as positive with NKX3.1, compared with 79 with PASD and 75 with Grocott. NKX3.1 was significantly superior to Grocott but not to PASD (P<0.05). NKX3.1 was significantly more efficacious in leading to a positive diagnosis of esophageal candidiasis compared with Grocott, resulting in a significantly higher number of positive fragments per slide as well as the number of organisms per fragment, but not PASD. NKX3.1 will be useful to add to the stain armamentarium for Candida and possibly other fungal organisms.

Fluorescence In Situ Hybridization for Diagnosis of Whipple's Disease in Formalin-Fixed Paraffin-Embedded Tissue.

Whipple’s disease (WD) is a rare chronic systemic infection with a wide range of clinical symptoms, routinely diagnosed in biopsies from the small intestine and other tissues by periodic acid–Schiff (PAS) diastase staining and immunohistological analysis with specific antibodies. The aim of our study was to improve the pathological diagnosis of WD. Therefore, we analyzed the potential of fluorescence in situ hybridization (FISH) for diagnosing WD, using a Tropheryma (T.) whipplei-specific probe. 19 formalin-fixed paraffin-embedded (FFPE) duodenal biopsy specimens of 12 patients with treated (6/12) and untreated (6/12) WD were retrospectively examined using PAS diastase staining, immunohistochemistry, and FISH. 20 biopsy specimens with normal intestinal mucosa, Helicobacter pylori, or mycobacterial infection, respectively, served as controls. We successfully detected T. whipplei in tissue biopsies with a sensitivity of 83% in untreated (5/6) and 40% in treated (4/10) cases of WD. In our study, we show that FISH-based diagnosis of individual vital T. whipplei in FFPE specimens is feasible and can be considered as ancillary diagnostic tool for the diagnosis of WD in FFPE material. We show that FISH not only detect active WD but also be helpful as an indicator for the efficiency of antibiotic treatment and for detection of recurrence of disease when the signal of PAS diastase and immunohistochemistry lags behind the recurrence of disease, especially if the clinical course of the patient and antimicrobial treatment is considered.

Correlation between histopathologic features and likelihood of identifying superficial dermatophytosis with periodic acid Schiff-diastase staining: a cohort study.

Periodic acid Schiff-diastase (PAS-D) staining is used routinely in dermatopathology. Conventionally, parakeratosis or intracorneal neutrophils justify use of PAS-D stain to identify dermatophytosis. We aimed to investigate the histopathological features of superficial dermatophytosis. Skin biopsies with PAS-D stain were retrospectively reviewed by a blinded dermatopathologist. Histopathologic findings in cases with dermatophytosis were compared with those without. We studied 110 cases, of which 63 were PAS-D positive for dermatophytes. Dermatophytes were identified on hematoxylin and eosin (H&E) staining in 49 (77.8%) of the PAS-D-positive cases. Diffuse compact orthokeratosis (79.37%, p < 0.001) and focal parakeratosis (65.08%, p = 0.03) were significantly more frequent in dermatophytosis. Intracorneal neutrophils and spongiosis were not significant findings. Fungal elements were frequently observed within compact orthokeratosis (62.0%) and parakeratosis (31.8%) but rarely within intracorneal pustules (12.7%). In some dermatophyte cases, intracorneal neutrophils (47.6%) or parakeratosis (17.5%) were absent. Compact orthokeratosis or focal parakeratosis justifies PAS-D staining, whereas the absence of intracorneal neutrophils or parakeratosis is insufficient to exclude dermatophytosis. Results indicate that having a low threshold to perform PAS-D stain is justified. PAS-D stain may be deferred until H&E slides have been evaluated.

Periodontal health status and implication of periodic acid-Schiff diastase - a key in exfoliative cytology among diabetics mellitus patients: A case-control study.

Objectives:The objectives of the study are: (i) To determine if periodic acid–Schiff (PAS)-Diastase is an effective tool to diagnose DM noninvasively, (ii) to use three different types of staining procedures to identify the staining pattern over the exfoliated cells of normal and diabetic patients, (iii) to study the periodontal health status of DM and normal subjects.Materials and Methods:Basic questions regarding the demographic data were asked, following which community periodontal index (CPI) was recorded. Oral smears were collected from clinically normal buccal mucosa of 150 patients using wooden tongue spatulas. The subjects were asked to gargle their mouth with water and the wooden spatula was scraped at the site from buccal mucosa. The smears were spread evenly on a dry clean glass slide and fixed immediately with absolute ethyl alcohol. Three slides were prepared for each of the patients. PAS, PAS with diastase digestion, and Papanicolaou staining were performed and examined under a microscope.Results:Average CPI for study subjects was 3.2 and control subjects was 2.1. The average loss of attachment was 1.1 in the study group and 0.2 in the control group (P - 0.00) and the result are statistically highly significant. All the 150 cases (100%) were positive for PAS staining, while PAS-diastase (PAS-D) staining showed positivity only for normal subjects and was negative (100%) in the diabetic group (P - 0.00) and the result are statistically highly significant.Conclusions:Results of our study showed that exfoliative cytology of the oral cavity, when stained with PAS and PAS-D, can be used as an effective screening and diagnostic tool for DM patients.

Histopathological, Ultrastructural and Apoptotic Changes in Diabetic Rat Placenta.

The exchange of substances between mother and fetus via the placenta plays a vital role during development. A number of developmental disorders in the fetus and placenta are observed during diabetic pregnancies. Diabetes, together with placental apoptosis, can lead to developmental and functional disorders. Histological, ultrastructural and apoptotic changes were investigated in the placenta of streptozotocin (STZ) induced diabetic rats. Animal experimentation. In this study, a total of 12 female Wistar Albino rats (control (n=6) and diabetic (n=6)) were used. Rats in the diabetic group, following the administration of a single dose of STZ, showed blood glucose levels higher than 200 mg/dL after 72 hours. When pregnancy was detected after the rats were bred, two pieces of placenta and the fetuses were collected on the 20(th) day of pregnancy by cesarean incision under ketamine/ xylazine anesthesia from in four rats from the control and diabetic groups. Placenta tissues were processed for light microscopy and transmission electron microscopy (TEM). Hematoxylin-eosin (HE) and periodic acid Schiff-diastase (PAS-D) staining for light microscopic and caspase-3 staining for immunohistochemical investigations were performed for each placenta. Electron microscopy was performed on thin sections contrasted with uranyl acetate and lead nitrate. Weight gain in the placenta and fetuses of diabetic rats and thinning of the decidual layer, thickening of the hemal membrane, apoptotic bodies, congestion in intervillous spaces, increased PAS-D staining in decidual cells and caspase-3 immunoreactivity were observed in the diabetic group. After the ultrastructural examination, the apoptotic appearance of the nuclei of trophoblastic cells, edema and intracytoplasmic vacuolization, glycogen accumulation, dilation of the endoplasmic reticulum and myelin figures were observed. In addition, capillary basement membrane thickening, capillary endothelial cells chromatin condensation in the nucleus and corrugation of the nucleus were found. Diabetes causes histomorphometric, ultrastructural and apoptotic changes in rat placenta.

Undifferentiated (embryonal) sarcoma of liver.

A 7-year-old male was brought to our out-patient department by his father with complaint of pain in the abdomen for 10 days. The parents noticed a swelling in the right upper abdomen a day after the pain started. He denied any past history of fever, jaundice, gastrointestinal blood loss or trauma. Examination revealed a non-tender swelling in the right hypochondrium which moved with respiration. Blood work, including tumor markers, was within normal limits. Contrast-enhanced computed tomographic (CECT) scan of abdomen was carried out which showed a large, hypodense, peripherally enhancing lesion involving the right lobe of liver (Panels A,B). The child underwent right hepatectomy which revealed a 15 cm × 12 cm sized solid exophytic tumor adhered to anterior abdominal wall and diaphragm focally. Grossly, the tumor showed a variegated appearance (Panel C). Histopathological examination revealed malignant nuclear features with cellular polymorphism (Panel D). Intracytoplasmic hyaline globules were also seen (Panel E) which were periodic acid-schiff (PAS) positive and diastase resistant. In addition, positivity for vimentin (Panel F), alpha-1-antitrypsin (Panel G) and Pan-CK (Panel H) was seen consistent with the diagnosis of undifferentiated (embryonal) sarcoma. The child had an uneventful recovery, being discharged 7 days after the surgery.

Periodic Acid Schiff-Diastase as a key in Exfoliative cytology in diabetics: A pilot study.

Background:Diabetes mellitus is the fifth most common chronic condition and the sixth most frequent cause of death among the elderly. The objective of this research was to develop a new method for diabetes diagnosis by analysis of the glycogen content of the oral epithelial cells.Materials and Methods:Ten control subjects and ten diabetic patients (study group) were taken, four oral smears for both control and study group from the buccal mucosa were taken and stained with hematoxylin and eosin stain, Papanicolaou (PAP) stain, periodic acid Schiff (PAS) stain and PAS-Diastase (PAS-D) stain.Results:The results showed that in the diabetic group: (i) The epithelial cells stained with PAP stain exhibited figures of binucleation and occasional karyorrhexis, (ii) the epithelial cells treated with PAS-D showed that glycogen containing cells did not take up the stain as compared to the other cells.Conclusion:The results associated with clinical and histological observations suggest that diabetes mellitus can produce alterations of oral epithelial cells as well as in their glycogen content.

Twenty-first-century oral hairy leukoplakia—a non–HIV-associated entity

This study presents the clinicopathologic features of a series (N = 35) of patients with non-human immunodeficiency virus (HIV)-associated oral hairy leukoplakia (OHL). Patients with non-HIV-associated OHL were identified from three centers. Epstein-Barr virus infection was demonstrated by using EBV early ribonucleic acid in situ hybridization. The presence of Candida co-infection was evaluated by diastase periodic acid-Schiff staining. The clinical features were determined by review of the medical records. Twenty-eight patients had intercurrent respiratory problems requiring long-term steroid inhaler use, four suffered from autoimmune diseases requiring immunosuppressant therapy, and four had diabetes. The majority of lesions were located on the tongue, and 24 showed evidence of Candida co-infection. In the twenty-first century, the presence of OHL should not be regarded as pathognomic for HIV infection or significant systemic immunosuppression. Local and systemic immunosuppression, in the form of steroid inhaler use, is a risk factor for the development of OHL.

Young Nepalese Male With Jaundice

Introduction: Pancreatic acinar cell carcinoma is a rare malignancy accounting for less than 1% of all pancreatic tumors. Given its rarity, diagnosis is often difficult and delayed. There are no current standard treatment protocols and surgical resection is the only curative measure. This cancer is primarily seen in older males. However, we report a case of acinar cell carcinoma in a young male from Nepal who presented with jaundice. Case Report: A 30-year-old Nepalese male with no significant past medical history who presented with jaundice, generalized pruritus, dark urine, and a 34-pound weight loss in 1.5 months. He presented with elevated serum liver enzymes. An abdominal ultrasound was performed, which showed fatty liver, distended gallbladder containing large amounts of sludge, and dilated common bile duct with sludge. A subsequent MRCP confirmed a dilated distal common bile duct along with a filling defect, 2.0 cm from ampulla of Vater with no other significant abnormalities. His initial ERCP showed a stricture in the common bile duct, a sphincterotomy was performed, and a stent was placed. Cytology was nondiagnostic and only showed atypical glandular cells. A CT did not show any other lesions or gross abnormalities. Subsequent endoscopic ultrasound (EUS) showed a well-demarcated mass in the lower third of the common bile duct with no significant pathology in the pancreas. Another ERCP was performed, which showed a dilated common bile duct with distal stricture that was restented and nondiagnostic biopsies. The patient’s laboratory values showed a slightly elevated CA 19-9, normal CEA, negative ANA, and normal IgG4, making autoimmune pancreatitis unlikely. The patient underwent a pylorus-sparing pancreaticoduodenectomy (partial Whipple resection), abdominal lymphadenectomy, and cholecystectomy. Pathology showed a 3-cm high-grade acinar cell carcinoma in the pancreatic head. The final staging of the tumor was T3N1M0 with 2 out of 5 resected abdominal lymph nodes being positive for metastasis. All resection margins were negative. Tumor cells were positive for Cam 5.2, AE1/AE3 and anti-chymotrypsin, making it consistent with acinar cell carcinoma in addition to positive PAS (periodic acid-Schiff-diastase) staining of cytoplasmic granules. The patient was started on a course of radiation with 5 fluorouracil, 3 months after surgery, followed by chemotherapy with Gemzar. After 1 month post-adjuvant radiation and chemotherapy, the patient’s CA 19-9 decreased. At 12 months’ follow-up, the patient is doing well and is on oral medications for brittle diabetes. The patient’s follow-up PET scan was negative.