PAS-positive Granules Research Articles

Is PAS Stain Necessary to Exclude Whipple Disease in Duodenal Biopsies?

Objective. Whipple disease, caused by Tropheryma whipplei, is a rare infectious condition primarily presenting with malabsorptive diarrhea. Small bowel biopsies typically reveal foamy macrophages containing periodic acid-Schiff-positive/diastase (PAS/D) resistant T. whipplei bacilli, and PAS(D) staining is occasionally requested by clinicians in duodenal biopsies, which are often histologically unremarkable. The yield of PAS(D) staining in such biopsies has never been reported to our knowledge. Methods. The anatomic pathology database was searched for all specimens of duodenal biopsies stained with PAS(D) from 1993 to 2021. Specimens were categorized by the following histomorphologic features: unremarkable, nonspecific changes, or expansion of the lamina propria by aggregates of foamy histiocytes. Follow-up information was collected, including microbiologic confirmatory testing. Results. There was a total of 193 specimens of duodenal biopsies stained with PAS(D). Biopsies lacking foamy histiocytes on H&E (n = 158) were never PAS-positive. Thirteen biopsies contained PAS-positive histiocytes; 9 out of the 13 PAS-positive specimens were subsequently confirmed to be T. whipplei. Of the 193 specimens, 124 specimens had a clinical request for PAS(D) staining. Only 3 of the 124 (2.4%) specimens showed foamy histiocytes containing PAS-positive granules, all of which were confirmed positive for T. whipplei. Conclusion. PAS(D) staining is unnecessary to exclude Whipple disease in duodenal biopsies without foamy macrophage aggregates, regardless of clinical suspicion. Clinical suspicion-driven biopsies often yield negative results for Whipple disease.

A172 A CLASSIC CASE OF WHIPPLE'S DISEASE

Abstract Background Whipple’s disease is a systemic disorder caused by the Gram-positive bacillus Tropheryma whippelii. It is classically defined by the four cardinal clinical manifestations of arthralgias, weight loss, diarrhea and abdominal pain. Classic Whipple’s disease is considered extremely rare, with an estimated global incidence of 30 cases per year, though this may be an underestimate. Aims To present a case of classic Whipple’s disease, so as to raise awareness and clinical suspicion for this rare condition. Methods We performed a retrospective chart review of a case of classic Whipple’s disease diagnosed in Saint John, NB, in 2020. Results A 55-year-old female presented to hospital with intractable weakness and diarrhea. Her history was notable for several years of right leg weakness, myalgias and low-grade fevers. More acutely, she presented to hospital due to worsening secretory diarrhea resulting in 15kg weight loss. Initial investigations were remarkable for an elevated C-reactive protein. The patient was admitted for coordinated evaluation for possible rheumatologic and infectious causes. As part of that workup, colonoscopy was performed. Colonoscopy revealed an abnormal ileum, with villi described as edematous with a mosaic pattern alternating between erythematous and “white tipped”. On histologic assessment, ileal biopsies showed macrophages with PAS-positive diastase resistant granules suspicious for Whipple’s disease. A follow-up esophagogastroduodenoscopy with duodenal biopsies was thus performed to gain further tissue for confirmatory testing. The duodenal mucosa was also noted to have a mosaic pattern with hypertrophic villi alternating between pale and erythematous in appearance. Duodenal biopsies were taken and sent for PCR testing. This testing was positive for T. whippelii, confirming the diagnosis of Whipple’s disease. The patient was initiated on long-term antibiotic therapy with ceftriaxone transitioning to trimethoprim-sulfamethoxazole prior to discharge. She unfortunately re-presented one month post-discharge with recurrent fevers, malaise and arthralgias. Given suspicion that her disease may be resistant to trimethoprim-sulfamethoxazole, she was transitioned to doxycycline and hydroxychloroquine as second-line therapy. She clinically improved on this regimen, and was able to be de-escalated to long-term doxycycline monotherapy after one year. At a recent follow-up, nearly three years from her initial diagnosis, she was doing well with essentially no ongoing symptoms of her Whipple’s disease. Conclusions Whipple’s disease is a rare infection presenting with intractable diarrhea and abdominal discomfort. Our case highlights the excellent clinical response that can be achieved with successful diagnosis and management of this otherwise debilitating disease. Funding Agencies None

Phenotypic description of Egyptian endemic centipedes, genus Scolopendra Linnaeus, 1758 with a histological study of its venom glands

Centipedes are widely distributed over all continents. They have significant medicinal value and high toxicity, making them an intriguing subject for evolutionary research. The current study aims to provide the first comprehensive analysis of the morphology, description, and distribution preferences of the Egyptian Scolopendromorpha centipede fauna. According to surveillance research in Egypt, two Scolopendra species have been re-described. Scolopendra canidensNewport.1844 were discovered in only one location, whereas Scolopendra morsitans s Linnaeus,1758 was more abundant. Both venomous glands were histochemically investigated. This study is the first comparative report on the histology and histochemistry of the venom glands of the Egyptian centipede species. Both species’ glands are primarily composed of columnar secretory cells radially arranged side by side and open through pores in a central chitinous duct. Each secretory cell is enclosed in striated muscle fibers. The secretion of both glands takes the shape of small PAS-positive granules, suggesting the presence of neutral polysaccharides. According to this surveillance study, the most abundant species was Scolopendra morsitans s Linnaeus,1758. Scolopendra canidens canidensNewport.1844 were found only on the Northwest coast of Egypt. The findings also revealed that the analyzed species are comparable in terms of their venom morphology and fundamental chemical composition.

What could this dark lesion on a woman’s eyelid be?

What could this dark lesion on a woman’s eyelid be?

Coagulansin-A improves spatial memory in 5xFAD Tg mice by targeting Nrf-2/NF-κB and Bcl-2 pathway

The present study was designed to investigate the underlying molecular signaling of Coagulansin-A (Coag-A) as a therapeutic agent against Alzheimer’s disease (AD). Preliminarily, it exhibited a neuroprotective effect against H2O2-induced oxidative stress in HT-22 cells. The in vivo studies were performed by administering Coag-A (0.1, 1, and 10 mg/kg) intraperitoneally to 5xFAD transgenic (Tg) mouse model. Coag-A (10 mg/kg) significantly attenuated the cognitive decline compared to Tg mice group in the shallow water maze (SWM) and Y-maze test paradigms. The anti-aggregation potential of Coag-A was determined by performing Fourier transform-infrared (FT-IR) spectroscopy and differential scanning calorimeter (DSC) analysis in the prefrontal cortex (PFC) and hippocampal (HC) regions of mice brain. The FT-IR spectra demonstrated the inhibition of amyloid beta (Aβ) through a decrease in β-sheet aggregation, along with the inhibition of changes in the lipids, proteins, and phospholipids. The DSC analysis displayed a low-temperature exotherm associated with the reversible process of aggregation of soluble protein fractions prior to denaturation. Furthermore, Coag-A treatment displayed a regular density of granule cells in H&E stained sections, along with a reduced amyloid load and PAS-positive granules in all the regions of interest in mice brain. The real-time polymerase chain reaction (q-PCR), western blot and immunohistochemical (IHC) analysis demonstrated antioxidant, anti-inflammatory, and anti-apoptotic effect of Coag-A by enhancing the expression of nuclear factor erythroid-2-related factor (Nrf-2) and reducing nuclear factor kappa B (NF-κB) and Bax protein expression. In addition, Coag-A significantly increased the antioxidant enzymes and proteins level, along with a reduced pro-inflammatory cytokines production.

Comparative analysis of the olfactory organs in selected species of marine sharks and freshwater batoids

We present comparative analysis of the micro- and macro-morphology of the olfactory organ of six neoselachians living in different habitats: two freshwater benthopelagic batoids, Potamotrygon motoro and Potamotrygon hystrix (Potamotrygonidae) as well as four species of sharks occupying the outer continental shelves and upper slopes Mustelus mustelus (Trikidae) and Scyliorhinus canicula (Scyliorhinidae), and deep-water Galeus melastomus (Scyliorhinidae) and Etmopterus spinax (Etmopteridae). Our research used light and electron (TEM, SEM) microscopes as well as macroscopic observations. The position of the nostrils in relation to the mouth varied in these species but all of them possessed elongate olfactory rosettes with primary lamellae arranged over two rows. The types of primary lamellae were distinguished based on their arrangement on the opposite sides of the median raphe as well as morphology of secondary lamellae and distribution of sensory and non-sensory regions of olfactory epithelium lining the lamellae; these were different between batoids and sharks. Analysis of olfactory epithelium revealed the presence of microvillous olfactory sensory neurons as the only type of receptor cells in all studied species whereas ionocytes were recognized only in sharks. All species possessed one type of goblet cells (Alcian Blue or Periodic Acid Schiff positives) except for P. motoro in which both types of goblet cells were present. Moreover, apical parts of non-sensory microvillous cells that lined the basal parts of the primary lamellae, possessed PAS-positive mucous granules; these were present in both species of batoids. This analogous region was abundant in goblet cells in the species of sharks studied here. Results indicated that differences between freshwater and marine species as well as between marine species occupying various habitat zones include mainly nostrils position, density of occurrence of ionocytes and goblet cells within olfactory epithelium as well as biochemical character of mucus.

Brain Lesions in Aging Zoo-Housed Naked Mole-Rats (Heterocephalus glaber).

Naked mole-rats (NMRs) are common in the managed care of zoos and valuable models for aging research. Limited information on NMR neuropathology is available despite many studies regarding their aging physiology. Histologic sections of brain from 27 adult (5-27 years old) NMRs from 2 zoos were reviewed to determine presence or absence of lesions associated with advanced age in humans and other mammals. A majority (23/27; 85%) of NMR brains had cerebral cortical neuronal changes with rounded or angular neurons, cytoplasmic vacuoles containing pale yellow pigment, periodic acid-Schiff (PAS)-positive granules and green autofluorescence, compatible with lipofuscinosis. Less severe lesions were present in cerebellar Purkinje cells, medulla, and hippocampal neurons. The hypothalamic neuropil of all NMRs had scattered variably sized PAS-positive granules and 10 (37%) had larger round bodies consistent with corpora amylacea. The youngest NMRs, 5 to 7 years old, generally had minimal or no cerebrocortical lesions. Further studies will help understand brain aging in this long-lived species.

Invasive Mammary Carcinoma With Paneth Cell–Like Eosinophilic Cytoplasmic Granules: Is It an Entity?

Abstract Breast carcinomas with Paneth cell–like eosinophilic cytoplasmic granules are rare and commonly reported in association with acinic cell carcinoma or microglandular adenosis. Paneth cell–like differentiation has been described outside the GI tract, including lung, liver, pancreas, lacrimal glands, and prostate. Very little is known about the significance of these granules and whether they represent a metaplastic or possibly a therapy-related change. There are only 45 reported cases involving the breast, mostly individual case reports or 2 to 3 case series. Although the clinical experience is limited, most of the reported cases seem clinically indolent. Here we present a 74-year-old female with a mammographically detected 1.1-cm left breast retroareolar mass for which she underwent a biopsy and subsequent lumpectomy with axillary dissection. Histology revealed an invasive ductal carcinoma, with Paneth cell–like eosinophilic PAS-positive cytoplasmic granules in the background of extensive ductal carcinoma in situ (DCIS). Similar granules were seen in some ducts involved by DCIS. Invasive carcinoma and DCIS were present throughout the 5.5-cm lumpectomy beyond the grossly measured 4.0-cm mass. There was extensive lymphovascular invasion and tumor was present at all the surgical resection margins. Twelve of the 13 axillary lymph nodes had macrometastases. The tumor was ER and PR positive and HER2 negative. Clinically, there were extensive bony metastases involving the skull, spine, sternum, iliac bones, and femur. There were also multiple lung nodules and mediastinal lymph nodes that were positive for metastasis on biopsy. The patient is still undergoing chemotherapy and radiotherapy 6 months following surgical excision. Our case demonstrates the heterogeneity of this entity as the clinical course in our patient has been very aggressive. A collaborative effort must be established to compile a larger case series to better our understanding of this increasingly reported histological observation and whether it represents a distinct entity.

New Observations Allowing the Differentiation of Late Asexual Stages of Cystoisospora canis from Developing Microgamonts in the Intestines of Experimentally Infected Dogs

The coccidian parasite Cystoisospora canis (syn. Isospora canis) can cause clinical disease in dogs. Three generations of meronts are reported to occur in the small intestine of experimentally infected dogs before gametogony and oocyst formation. Oocyst excretion in the feces occurs at 9 to 11 days post-inoculation (PI). We examined the late asexual and sexual development of C. canis in 2 dogs necropsied 10 days after oral inoculation with 100,000 sporulated C. canis oocysts; both dogs had excreted oocysts 9 days PI. Asexual and sexual stages were seen in the lamina propria, throughout the small intestine in sections stained with hematoxylin and eosin from both dogs. In other studies of the C. canis life cycle, little attention has been given to distinguishing the last asexual generation of meronts and early microgamonts that can appear similar due to their stage of maturation and both having multiple nuclei. Here we report newly identified features of developing meronts and microgamonts and their distinction from each other by using sections processed using the periodic acid-Schiff (PAS) reaction. Using this method, we demonstrated that PAS-positive granules could be used to identify microgamonts and differentiate them from developing meront stages. These findings will aid pathologists and others in properly identifying coccidial parasites, in determining the cause of microscopic lesions in intestinal tissue, and in accurately identifying etiological agents.

Re-evaluation of merogony of a Cystoisospora ohioensis-like coccidian and its distinction from gametogony in the intestine of a naturally infected dog.

Four species of Cystoisospora, C. canis, C. ohioensis, C. neorivolta and C. burrowsi are described from feces of dogs. Of these, the oocysts of C. canis are the largest and easily distinguished from the remaining three species. Oocysts of C. ohioensis, C. neorivolta and C. burrowsi are difficult to distinguish because of overlap in their sizes. However, based on endogenous developmental stages, C. ohioensis is distinct from C. neorivolta and C. burrowsi because its endogenous stages are confined to surface epithelium of intestine whereas endogenous stages of C. neorivolta and C. burrowsi are predominantly in the lamina propria. There are uncertainties regarding the endogenous stages of C. neorivolta and C. burrowsi and there is no way now to determine whether C. burrowsi and C. neorivolta are different parasites; therefore, these are referred as C. ohioensis-like organisms. Additionally, mode of division of asexual stages of coccidia of dogs is largely unknown and ultrastructural studies are lacking. In the present study, development of asexual and sexual stages of a C. ohioensis-like organism in a naturally infected dog is described by light microscopy and by transmission electron microscopy. Merozoites divided by endodyogeny/merogony. Meronts were crescent/merozoite-shaped and contained a maximum of eight nuclei. A distinctive feature of merozoites was the presence of many PAS-positive amylopectin granules that were absent or rare in immature microgamonts making it possible to distinguish them.

Histopathological evaluation of oral membranous substance in bedridden elderly persons without oral intake in Japan.

The aim of this study was to clarify by histopathological examination the origin of oral membranous substances deposited on the palate, tongue, buccal mucosa and teeth. Several investigators have reported membranous substances deposited in the mouths of bedridden elderly persons requiring nursing care without oral intake. However, the precise nature and origin of the substances are poorly understood. Sixty-nine specimens were taken from the oral cavity of bedridden patients, that is, the palate, dorsum of the tongue, the cheek and teeth. Sections were stained with haematoxylin and eosin stain, alcian-blue and periodic acid-Schiff stain (AB-PAS) and antibodies for pankeratin (AE1AE3) and leukocyte common antigen (LCA). All specimens showed a film-like nature coloured from tan to white, accompanied by a mucous substance. Histologically, specimens of all sites had a similar feature of the combination of basophilic amorphous and eosinophilic lamellar features. The basophilic substance was positive for AB-PAS, and PAS-positive glycogen granules were also noted in the lamellar structure. Immunochemistry revealed various degrees of pankeratin positive substance and LCA-positive inflammatory cell infiltration. The oral membranous substance was composed of keratin and mucin with inflammation. These results suggest that the deposition of the oral membranous substance is a pathological condition or oral mucositis caused by dry mouth.

Pathological Findings in the Pituitary Glands of Dogs and Cats.

With the exception of classic functional adenomas in dogs and horses, pituitary lesions are infrequently described in the veterinary literature. Approximately 10% of pituitary glands from asymptomatic humans contain abnormalities, but the equivalent proportion in small animals is unknown. Pituitary glands from 136 dogs and 65 cats collected during routine necropsies were examined to determine the prevalence of pituitary lesions and their histopathological diagnosis. Lesions were characterized in sections stained with hematoxylin and eosin, periodic acid-Schiff (PAS), Gordon and Sweet's and reticulin stains, and immunohistochemistry for adrenocorticotropic hormone (ACTH), growth hormone, melanocyte stimulating hormone-α, and prolactin. Pituitary abnormalities were identified in 36 of 136 (26.4%) dogs and 10 of 65 (15.3%) cats. Cystic changes were the most common lesion, occurring in 18 (13.2%) dogs and 8 (12.3%) cats. Pituitary neoplasia was detected in 14.1% (12/85) of middle-aged and old dogs; 1 (1.5%) cat had pituitary nodular hyperplasia. PAS and reticulin stains helped differentiate ACTH-immunoreactive adenomas from hyperplastic nodules: adenomas contained PAS-positive intracytoplasmic granules and loss of the normal reticulin network. One dog had a pituitary carcinoma with infiltration into the thalamus. Other pituitary abnormalities included secondary metastases (2 dogs) and hypophysitis (4 dogs, 1 cat). In most cases, the lesion appeared to be subclinical and could be considered incidental, whereas clinical manifestations were apparent in only 4 dogs (2.9%) and none of the cats with pituitary lesions. Pituitary abnormalities are common in dogs and cats, and their clinical relevance requires further investigation.

Granular cell tumors of the tongue: fibroma or schwannoma

BackgroundGranular cell tumors are benign lesions that typically occur in the oral cavity, but can also be found in other sites. However, the characteristics of these tumors are unclear. Thus, the present study aimed to investigate the immunohistological characteristics of these tumors of the tongue.MethodsSeven patients were treated for granular cell tumors of the tongue at our institution during 2003–2017. Paraffin-embedded specimens were available for all cases; thus, retrospective immunohistochemical analyses were performed.ResultsAll cases exhibited cytoplasmic acidophilic granules in the muscle layer of the tumor. Both the normal nerve cells and tumor cells also stained positive for PGP9.5, NSE, calretinin, and GFAP. A nucleus of tumor cells was typically present in the margin. The PAS-positive granules were also positive for CD68 (a lysozyme glycoprotein marker). Various sizes of nerve fibers were observed in each tumor, and granular cells were observed in the nerve fibers of a representative case.ConclusionsBased on our immunohistological findings, granular cell tumors may be derived from Schwann cells, and the presence of CD68 indicates that Wallerian degeneration after nerve injury may be a contributor to tumor formation. Thus, a safe surgical margin is needed to detect the infiltrative growth of granular cell tumors.

P.215 - Sensitivity and specificity of the PAS positive lymphocyte vacuoles in the diagnostic approach to late onset Pompe disease

The autosomal recessive Pompe disease, which is caused by deficiency of the lysosomal enzyme acid α-glucosidase (GAA), manifests in the very severe classic-infantile and in milder course late-onset forms (LOPD). Since an effective Enzyme Replacement Therapy is available, a cheap, rapid, sensitive and specific screening test is crucial to early identify homozygous affected and heterozygous carriers. Aim of this study is to verify sensitivity and specificity of the quantitative assessment of PAS-positive granules in lymphocytes to recognize homozygous and heterozygous GAA mutated individuals. Number of lymphocytes containing PAS positive granules as well as the number of granules in each lymphocyte were assessed on periodic acid- Schiff (PAS) stained blood smears from 72 adult patients with persistent hyperckemia and/or muscle weakness, 7 patients with genetically confirmed LOPD, and 13 patients with heterozygous GAA mutations. Percentage of affected lymphocytes and mean number of granula per lymphocyte were calculated for all patients. Accuracy of the test was evaluated by ROC curve. Eleven out of 72 patients with hyperckemia and/or muscular weakness showed more than 4 PAS-positive lymphocytes (6 new diagnosis of LOPD, 4 Glycogen storage myopathies different from Pompe, and 1 caveolinopathy). All the 22 LOPD patients and, surprisingly, also all the 13 GAA-mutation carriers presented number of PAS-positive granules higher than in other myopathies. PAS-positive lymphocytes analysis disclosed a 97%sensitivity and a 94% specificity in recognizing homozygous and heterozygous GAA mutations. Quantification of PAS-positive lymphocytes in blood smears should be routinely used as first level screening test for Pompe disease.

Origin of Rodlet Cells and Mapping Their Distribution in Ruby-Red-Fin Shark (Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light, Immunohistochemistry and Ultrastructure Study

The present study describes for the first time histology, histochemistry, and ultrastructure of all rodlet stages. By ultrastructure, we identified rodlet progenitors in the stroma of the olfactory organ in Red-Fin shark. Rodlet progenitors were mesenchymal-like cells synthesis the primitive rodlet granules. Rodlet progenitors differentiated to vesicular rodlet cells, which was rich in vesicles and vacuoles, and accumulate intracellular fibrillar-like components. Granular rodlet cells were observed in the basal epithelia, contained premature rodlet granules and began to organize the fibrillar components of the rodlet capsule. Transitional rodlet cells continued to deposit the fibrils of the capsule and synthesis of rodlet granules. Mature rodlet cells were polarized bear-shape and had typical rodlet granules. Histochemistry showed rodlet cells had a wide range of staining affinities including carbohydrate, lipid, and protein staining. All stages of rodlet cells revealed the presence of PAS-positive granules in the cytoplasm of rodlet cells where, While the ruptured ones stained strongly with alcian blue, sliver stain and Sudan black B. Rodlet granule was positive for bromophenol blue Rodlet cells were positively immunostained against Matrix Metalloproteinase-9 (MPP-9) in all stages indicate invasion properties of rodlet cells in tissue. Conclusion rodlet cells originated from the stroma of the olfactory organ.

Characterization of Periodic Acid-Schiff-Positive Granular Deposits in the Hippocampus of SJL/J Mice.

Periodic acid-Schiff (PAS)-positive granular deposits in the hippocampus have been reported previously in certain inbred mouse strains such as C57BL/6 and the senescent-accelerated mouse prone-8. Here, we report for the first time that similar PAS-positive granules age dependently occur in SJL/J mice, a mouse strain, for instance, used for central nervous system disease research. Moreover, similar granules stained intensely positive with a polyclonal antibody directed against p75 neurotrophin receptor (p75(NTR)). Granular deposits were absent in young mice and developed with aging in CA1 and CA2 regions of the hippocampus. Interestingly, granules significantly diminished in SJL/J mice previously treated with cuprizone, a copper chelator, which is a useful model for toxic demyelination. The presented data support the idea that granules might be the result of an imbalance of redox-active metals and/or a dysregulation of complementary mechanisms that regulate their homeostasis in astrocyte-neuron coupling, respectively. It remains to be determined whether the unsuspected immunoreactivity for p75(NTR) represents a false-positive reaction or whether p75(NTR) is crucially involved in the pathogenesis of age-related hippocampal granular deposits in mice.

Light and electron microscopic study of the eyelids, conjunctiva-associated lymphoid tissue and lacrimal gland in Bilgorajska Goose (Anser anser).

Normal structure of the accessory organs of the eye is essential for normal eye physiology. Among the most important accessory organs of the eye are the eyelids, the conjunctiva-associated lymphoid tissue (CALT) and the lacrimal gland (LG). The aim of this study was to demonstrate the histological structure of the eyelids and LG by histochemical and ultrastructural analysis. The study was performed on 13 adult female Bilgorajska geese. Eyelid samples were stained with the Alcian blue (AB pH 2.5) and periodic acid-Schiff (PAS) methods. Staining methods used for LG were AB pH 2.5, aldehyde fuchsin (AF), PAS and Hale's dialysed iron (HDI). Within the connective tissue of the eyelids, well-developed, diffuse, CALT follicles were observed, mostly under the conjunctival epithelium. Numerous lymphocytes were present within loose connective tissue. Staining of the eyelids with the PAS method demonstrated the presence of goblet cells of a mucous nature, and AB pH 2.5 staining indicated the presence of sulfated acid mucopolysaccharides. PAS staining of LG revealed the presence of secretory cells containing weakly PAS-positive granules. All epithelial cells of the corpus glandulae and the duct systems reacted positively to AB pH 2.5. HDI staining detected the presence of carboxylated acid mucopolysaccharides. Transmission electron microscopy investigations revealed two types of secretory epithelial cells in LG. Both types of LG cells contained drop-like secretory vesicles of different sizes with low or high electron density in cytoplasm, as well as small and large lipid vacuoles, and numerous small primary lysosomes.

Detecting glycogen in peripheral blood mononuclear cells with periodic acid schiff staining.

Periodic acid Schiff (PAS) staining is an immunohistochemical technique used on muscle biopsies and as a diagnostic tool for blood samples. Polysaccharides such as glycogen, glycoproteins, and glycolipids stain bright magenta making it easy to enumerate positive and negative cells within the tissue. In muscle cells PAS staining is used to determine the glycogen content in different types of muscle cells, while in blood cell samples PAS staining has been explored as a diagnostic tool for a variety of conditions. Blood contains a proportion of white blood cells that belong to the immune system. The notion that cells of the immune system possess glycogen and use it as an energy source has not been widely explored. Here, we describe an adapted version of the PAS staining protocol that can be applied on peripheral blood mononuclear immune cells from human venous blood. Small cells with PAS-positive granules and larger cells with diffuse PAS staining were observed. Treatment of samples with amylase abrogates these patterns confirming the specificity of the stain. An alternate technique based on enzymatic digestion confirmed the presence and amount of glycogen in the samples. This protocol is useful for hematologists or immunologists studying polysaccharide content in blood-derived lymphocytes.

Diffuse parenchymal form of malakoplakia in renal transplant recipient: a case report

Malakoplakia is an unusual chronic inflammatory disease related to prior urinary tract infection. It is characterized by the presence of macrophages with foamy cytoplasm exhibiting larger PAS positive inclusions that stain for calcium and iron. Malakoplakia affects renal allograft and is associated with severe morbidity. Herein, the authors report a new case of renal graft malakoplakia in a 23-year-old female patient. The patient received a living-related donor renal transplantation with a high immunological risk. Plasmapheresis and intravenous immunoglobulin (i.v. Ig) treatment, pre- and post-transplant, and induction with rabbit anti-thymocyte globulins were used due to presence of donor specific antibodies and positive B cross match by flow cytometry. The patient had an early urinary tract infection with a good outcome. On Day 36 post-transplant (PO), the patient returned to the clinic with fever, graft pain and acute renal dysfunction leading to hemodialysis. Escherichia coli (E. coli) was present in the blood and urine culture. At the time, the renal biopsy revealed numerous sheets of macrophages with foamy, eosinophilic cytoplasm showing several PAS positive granules and large inclusions that stained strongly with hematoxylin, calcium (von Kossa method) and iron (Prussian blue). The patient was diagnosed with malakoplakia related to a kidney transplant. Despite prolonged treatment with antibiotics, determined by a susceptibility test, the patient did not recover renal function and remained on dialysis.

Orbital alveolar soft-part sarcoma: clinico-pathological profiles, management and outcomes.

To study the clinico-radiological and clinic-pathological features of orbital alveolar soft-part sarcomas (ASPS). We retrieved 9 cases of orbital ASPS. Histopathological and immunohistochemical features and clinico-radiological findings were evaluated. Age at presentation varied from 1 to 31 years. Both, children and adults were affected. Proptosis was the most common clinical presentation. PAS-positive diastase-resistant crystals and granules were found in all cases. All patients received adjuvant radiotherapy. Two cases had positive surgical margins and were treated with additional adjuvant chemotherapy. None of the patients had recurrence or metastases till the last follow up. Early diagnosis with a multidisciplinary, multimodal approach of treatment is quintessential in achieving a long disease-free survival in patients with ASPS.