Use Of Hematoxylin Research Articles

Somatostatin receptor 2A expression in von Hippel-Lindau-related hemangioblastomas.

Central nervous system hemangioblastomas are the most prevalent manifestation of von Hippel-Lindau (VHL) disease and remain the main cause of mortality. Surgical resection is the primary treatment strategy, but is not always possible, and should be used as restrictively as possible. There is an unmet need for less invasive treatment strategies, such as targeted therapy. Expression of somatostatin receptor 2A (SSTR2A) in VHL-related hemangioblastomas has been described earlier, but the extent of expression in a larger population has yet to be determined. The authors hypothesize that a substantial subset of VHL-related hemangioblastomas show SSTR2A expression, which may serve as a potential new treatment target. Patients who were surgically treated for a VHL-related hemangioblastoma from 1990 until 2021 at the UMC Utrecht were included. Clinical data was derived from a clinical database. Tissue samples were histopathologically examined with use of hematoxylin and eosin staining, and immunohistochemical analysis of SSTR2A expression was performed. Forty-three tissue samples were obtained from 26 patients. Nine showed strong positivity for SSTR2A expression, whereas 13 showed moderate and 15 sparse expression. Three samples showed no expression of SSTR2A. The distribution showed right-skewedness favoring a strong expression. SSTR2A expression colocalized with endothelial markers and not with stromal cells. Additionally, within-patient variability for SSTR2A expression was described in 14 patients. SSTR2A is expressed in varying degrees in the majority of VHL-related hemangioblastomas. Future treatment with somatostatin analogues or even peptide receptor radionuclide treatment may be considered for SSTR2A-positive cases.

Dihydromyricetin ameliorates hepatic steatosis and insulin resistance via AMPK/PGC-1α and PPARα-mediated autophagy pathway

BackgroundDihydromyricetin (DHM), a flavonoid compound of natural origin, has been identified in high concentrations in ampelopsis grossedentata and has a broad spectrum of biological and pharmacological functions, particularly in regulating glucose and lipid metabolism. The objective of this research was to examine how DHM affected nonalcoholic fatty liver disease (NAFLD) and its underlying mechanisms involved in the progression of NAFLD in a rat model subjected to a high-fat diet (HFD). Additionally, the study examines the underlying mechanisms in a cellular model of steatohepatitis using palmitic acid (PA)-treated HepG2 cells, with a focus on the potential correlation between autophagy and hepatic insulin resistance (IR) in the progress of NAFLD.MethodsSD rats were exposed to a HFD for a period of eight weeks, followed by a treatment with DHM (at doses of 50, 100, and 200 mg·kg−1·d−1) for additional six weeks. The HepG2 cells received a 0.5 mM PA treatment for 24 h, either alone or in conjunction with DHM (10 µM). The histopathological alterations were assessed by the use of Hematoxylin–eosin (H&E) staining. The quantification of glycogen content and lipid buildup in the liver was conducted by the use of PAS and Oil Red O staining techniques. Serum lipid and liver enzyme levels were also measured. Autophagic vesicle and autolysosome morphology was studied using electron microscopy. RT-qPCR and/or western blotting techniques were used to measure IR- and autophagy-related factors levels.ResultsThe administration of DHM demonstrated efficacy in ameliorating hepatic steatosis, as seen in both in vivo and in vitro experimental models. Moreover, DHM administration significantly increased GLUT2 expression, decreased G6Pase and PEPCK expression, and improved IR in the hepatic tissue of rats fed a HFD and in cells exhibiting steatosis. DHM treatment elevated Beclin 1, ATG 5, and LC3-II levels in hepatic steatosis models, correlating with autolysosome formation. The expression of AMPK levels and its downstream target PGC-1α, and PPARα were decreased in HFD-fed rats and PA-treated hepatocytes, which were reversed through DHM treatment. AMPK/ PGC-1α and PPARα knockdown reduced the impact of DHM on hepatic autophagy, IR and accumulation of hepatic lipid.ConclusionsOur findings revealed that AMPK/ PGC-1α, PPARα-dependent autophagy pathways in the pathophysiology of IR and hepatic steatosis has been shown, suggesting that DHM might potentially serve as a promising treatment option for addressing this disease.Graphical

FDG PET/CT in Aseptic Splenic Abscess

HomeRadiologyRecently Published PreviousNext Reviews and CommentaryFree AccessImages in RadiologyFDG PET/CT in Aseptic Splenic AbscessWeiting Liu, Xiaohong Ou Weiting Liu, Xiaohong Ou Author AffiliationsFrom the Department of Nuclear Medicine, West China Hospital of Sichuan University, No. 37 Guoxue Alley, Chengdu 610041, People’s Republic of China.Address correspondence to X.O. (email: [email protected]).Weiting LiuXiaohong Ou Published Online:Mar 28 2023https://doi.org/10.1148/radiol.222955MoreSectionsPDF ToolsImage ViewerAdd to favoritesCiteTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinked In A 32-year-old woman presented with fever and arthralgia for 1 month. She had a past medical history of rheumatoid arthritis with 2 years of glucocorticoid treatment. At physical examination, splenomegaly was found. Her laboratory data showed leukocytosis (15.86 × 109/L; reference range, 3.5–9.5 × 109/L) and a high erythrocyte sedimentation rate. Serial blood cultures were negative. Intravenous antibiotics and corticosteroid therapy did not relieve her symptoms. Fluorodeoxyglucose (FDG)-PET/CT examination was performed, showing abnormal standardized uptake values in the spleen (Figure). Primary splenic lymphoma was suspected, and the patient underwent laparoscopic splenectomy. At pathologic examination (Figure), the diagnosis of aseptic splenic abscesses as a complication of rheumatoid arthritis was made. Corticosteroids and supportive treatment were then initiated, and her clinical condition gradually improved.Images in a 32-year-old woman presenting with fever and arthralgia for 1 month. She had a past medical history of rheumatoid arthritis with 2 years of glucocorticoid treatment. A fluorodeoxyglucose (FDG)-PET examination was performed. (A) Maximum intensity projection whole body image shows intense diffuse splenic uptake and moderate bone marrow uptake. (B) Axial-fused PET/CT abdominal image shows an enlarged spleen with heterogeneously increased FDG standardized maximum uptake value (standardized maximum uptake value,13.94; liver standardized maximum uptake value, 3.76). At pathologic examination, macroscopic examination of the spleen showed many nonconfluent white nodules (not shown). (C) With use of hematoxylin and eosin, a photomicrograph with 400× magnification shows numerous areas of necrosis surrounded by intense polymorphonuclear cell infiltration (arrows) and fibroblastic proliferation at the periphery of the splenic nodules.Download as PowerPointOpen in Image Viewer Splenic abscesses generally occur in patients with immune dysfunction (1), presenting as solitary or multifocal lesions. Aseptic abscesses are most commonly associated with nonspecific inflammatory bowel disease and other autoimmune diseases (spondyloarthropathy, rheumatoid arthritis, lupus erythematosus, and nodular arteritis), or autoinflammatory (synovitis, acne, pustulosis, hyperostosis, and osteitis) and neoplastic ones (myeloma, monoclonal gammopathy) (2). Histopathologic examination helps to exclude other conditions. The cornerstones of aseptic abscesses are mature polymorphonuclear neutrophils in the center of the splenic nodules, with histiocytes and single giant cells present at the periphery (2). It is unclear whether glucocorticoids could be a factor that promotes the formation of aseptic splenic abscesses. The presence of disseminated sterile splenic microabscesses with intense FDG uptake related to rheumatoid arthritis is rare.Disclosures of conflicts of interest: W.L. No relevant relationships. X.O. No relevant relationships.

The use of hematoxylin and eosin muscle staining and ImageJ as tools to assess the incidence and severity of white striping in chicken breast

The myopathy known as white striping (WS) increases deposits of fatty tissue in breasts (Pectoralis major) of high yielding broiler chickens. This condition threatens the poultry industry as it decreases consumers’ willingness to purchase. To compare macroscopic (visual scoring) and microscopic (histological staining) methods as tools to assess WS, samples were collected from a trial evaluating the effects of growth rate (fast or slow) and L-carnitine supplementation (0 or 100 mg/kg) on performance parameters of broilers. Chicken breasts (Pectoralis major; n=144) were biopsied on the left cranial ventral region. Histological slides were prepared and stained (hematoxylin-eosin; H&E), photographed, and analyzed using ImageJ software. Increased incidence and severity of WS was visually observed in fast growing birds (P<0.0001) and those supplemented with 100 mg/kg of L-carnitine (P=0.0348). Fast growth rates increased average cell area (P=0.0315) and percentage of adipose tissue (P=0.0007), while cell count was higher in slow growing birds (P=0.0171). A significant correlation (r=0.2375; P=0.0043) was found between visual assessment of adipose tissue and percentage determined microscopically. Although it was possible to determine presence and severity of WS by using H&E staining, this technique is labor intensive and costly relative to subjective visual assessment, which is comparatively more resource efficient.

Expression Profiles of GILZ and Annexin A1 in Human Oral Candidiasis and Lichen Planus.

Adrenal glands are the major source of glucocorticoids, but recent studies indicate tissue-specific production of cortisol, including that in the oral mucosa. Both endogenous and exogenous glucocorticoids regulate the production of several proteins, including the glucocorticoid-induced leucine zipper (GILZ) and Annexin A1, which play important roles in the regulation of immune and inflammatory responses. Common inflammation-associated oral conditions include lichen planus and candidiasis, but the status of GILZ and Annexin A1 in these human conditions remains to be established. Accordingly, archived paraffin-embedded biopsy samples were subjected to immunohistochemistry to establish tissue localization and profile of GILZ and Annexin A1 coupled with the use of hematoxylin–eosin stain for histopathological assessment; for comparison, fibroma specimens served as controls. Histopathological examination confirmed the presence of spores and pseudohyphae for oral candidiasis (OC) specimens and marked inflammatory cell infiltrates for both OC and oral lichen planus (OLP) specimens compared to control specimens. All specimens displayed consistent and prominent nuclear staining for GILZ throughout the full thickness of the epithelium and, to varying extent, for inflammatory infiltrates and stromal cells. On the other hand, a heterogeneous pattern of nuclear, cytoplasmic, and cell membrane staining was observed for Annexin A1 for all specimens in the suprabasal layers of epithelium and, to varying extent, for inflammatory and stromal cells. Semi-quantitative analyses indicated generally similar fractional areas of staining for both GILZ and Annexin A1 among the groups, but normalized staining for GILZ, but not Annexin A1, was reduced for OC and OLP compared to the control specimens. Thus, while the cellular expression pattern of GILZ and Annexin A1 does not differentiate among these conditions, differential cellular profiles for GILZ vs. Annexin A1 are suggestive of their distinct physiological functions in the oral mucosa.

Hypoxic hUCMSC-derived extracellular vesicles attenuate allergic airway inflammation and airway remodeling in chronic asthma mice

BackgroundAs one of the main functional forms of mesenchymal stem cells (MSCs), MSC-derived extracellular vesicles (MSC-EVs) have shown an alternative therapeutic option in experimental models of allergic asthma. Oxygen concentration plays an important role in the self-renewal, proliferation, and EV release of MSCs and a recent study found that the anti-asthma effect of MSCs was enhanced by culture in hypoxic conditions. However, the potential of hypoxic MSC-derived EVs (Hypo-EVs) in asthma is still unknown.MethodsBALB/c female mice were sensitized and challenged with ovalbumin (OVA), and each group received PBS, normoxic human umbilical cord MSC-EVs (Nor-EVs), or Hypo-EVs weekly. After treatment, the animals were euthanized, and their lungs and bronchoalveolar lavage fluid (BALF) were collected. With the use of hematoxylin and eosin (HE), periodic acid-Schiff (PAS) and Masson’s trichrome staining, enzyme-linked immune sorbent assay (ELISA), Western blot analysis, and real-time PCR, the inflammation and collagen fiber content of airways and lung parenchyma were investigated.ResultsHypoxic environment can promote human umbilical cord MSCs (hUCMSCs) to release more EVs. In OVA animals, the administration of Nor-EVs or Hypo-EVs significantly ameliorated the BALF total cells, eosinophils, and pro-inflammatory mediators (IL-4 and IL-13) in asthmatic mice. Moreover, Hypo-EVs were generally more potent than Nor-EVs in suppressing airway inflammation in asthmatic mice. Compared with Nor-EVs, Hypo-EVs further prevented mouse chronic allergic airway remodeling, concomitant with the decreased expression of pro-fibrogenic markers α-smooth muscle actin (α-SMA), collagen-1, and TGF-β1-p-smad2/3 signaling pathway. In vitro, Hypo-EVs decreased the expression of p-smad2/3, α-SMA, and collagen-1 in HLF-1 cells (human lung fibroblasts) stimulated by TGF-β1. In addition, we showed that miR-146a-5p was enriched in Hypo-EVs compared with that in Nor-EVs, and Hypo-EV administration unregulated the miR-146a-5p expression both in asthma mice lung tissues and in TGF-β1-treated HLF-1. More importantly, decreased miR-146a-5p expression in Hypo-EVs impaired Hypo-EV-mediated lung protection in OVA mice.ConclusionOur findings provided the first evidence that hypoxic hUCMSC-derived EVs attenuated allergic airway inflammation and airway remodeling in chronic asthma mice, potentially creating new avenues for the treatment of asthma.

Evaluation of Explanted CorMatrix Tyke Extracardiac Patches in Infants With Congenital Heart Disease

Animal data demonstrate that intracardiac patches of decellularized porcine small intestine submucosa (CorMatrix; CorMatrix Cardiovascular, Atlanta, GA) become repopulated with native cells, suggesting the possibility of a substrate for regenerative tissue in humans. Although human studies have not demonstrated this regenerative property with intracardiac patches, it is possible that other environments may better promote native cell repopulation. We report a prospective series of explanted CorMatrix extracardiac patches placed in infants with congenital heart disease. CorMatrix Tyke patches were used in pulmonary artery patch closure during the Norwood procedure. The patch material was explanted as part of the hemi-Fontan procedure. Specimens were analyzed with the use of hematoxylin and eosin, Movat pentachrome, and trichrome stains. Of the 11 implantations, 9 specimens were explanted. One patient did not survive to hemi-Fontan procedure. One patient's patch was removed, but not analyzed. Acellular material, chronic inflammation, fibrosis, and foreign body giant cell reaction was seen in all explanted patches. No explanted CorMatrix Tyke material demonstrated evidence of ingrowth of native endothelial tissue at a median of 4.9 months invivo. CorMatrix Tyke patch material, placed as a pulmonary artery patch in an extracardiac position, remained pliable and mostly free of calcification. However, these patches did not show evidence of native endothelial tissue at a median of 4.9 months invivo.

UTILIZATION OF 1% OF METHYLENE BLUE IN STAINING HISTOPATHOLOGICAL PREPARATIONS AT ANATOMIC PATHOLOGY LABORATORY

Tissue staining using hematoxylin-eosin (HE) is a standard method of histopathological staining. The tissue staining is hampered when there is no hematoxylin reagent in laboratory. Therefore, other reagents are needed that can replace the use of hematoxylin. Methylene blue is a basic dyes that interact with cell nuclei which has a negative ionic charge of the tissue. It can be used as an alternative nuclei staining. This study aims to evaluate the use of 1% of methylene blue in cell nuclei staining in histopathological preparations. The research sample were 15 pathology preparations which were randomly selected including breast cancer, cervical cancer and ovarian cancer in the bank of sampel at anatomical pathology laboratory of RSUD Dr. Slamet Garut, Indonesia. The experiment showed that the methylene blue dyes yielded “worth” result (40%) and “poorly” result (60%). Further research can be carried out by modifying the pH of 1% of methylene blue reagent so that it can maximize the staining preparations results as good as those using hematoxylin.

To assess calretinin immunohistochemistry in Hirschsprung’s disease and allied disorders

Hirschsprung’s disease (HD) is the congenital aganglionosis of the human colon due to disruption of migration, imbalanced proliferation and differentiation of neural crest cell. The diagnosis and extent of resection for the management of HD depend on the sensitive and specific identification of ganglion cells. Negative immunohistochemical expression of calretinin, shown its superiority as diagnostic method for full thickness as well as partial thickness rectal suction biopsies. Total 86 colonic and rectal biopsies of 43 patients attending Sassoon hospital, Pune (between 2015-2017) with surgically resected specimen proven Hirschsprung’s disease cases and follow up cases of non HD cases were evaluated with both Haematoxylin and Eosin stain and calretinin IHC. Age group was 2 days to 7 year with having Male: Female ratio 2.91:1. Out of 43 cases, 26 were diagnosed as Hirschsprung’s Disease, 16 were not having Hirschsprung’s disease, 1 case of hypogangliosis. 13 out of 86 biopsy and 9 out of 43 cases show false positive result with use of Haematoxylin and Eosin stain only. Specificity and negative predictive value of H&E stain was 72% and 78.12% respectively with accuracy of 86%. Use of calretinin IHC gave 100% accuracy in all 86 biopsies studied including 17 partial thickness biopsies. Only use of Haematoxylin and Eosin stain will lead to significant false positive result leading to misdiagnosis and in turn wrong treatment. Loss of calretinin expression in aganglionic segment help in the interpretation of even partial biopsies and easy to interpret with improvement in diagnosis upto 100%.

Molecular analysis of H&E- and Papanicolau-stained samples-systematic review.

Molecular pathology allows the identification of causative agents in infectious diseases and detection of biomarkers important for prediction of disease susceptibility, diagnosis and personalized therapy. Accordingly, nucleic acid-based methods have gained a special role in clinical laboratories particularly to evaluate solid and hematological tumors. Extraction of nucleic acids is commonly performed in microdissected formalin-fixed paraffin-embedded (FFPE) or cytological samples that had been previously evaluated through the use of hematoxylin and eosin (H&E) or Papanicolau (Pap) stains, respectively. Although the effect of both stains on nucleic acids integrity has been explored by several authors, the results are not consistent and require further examination. Accordingly, the goal of this review was to assess the influence of H&E and Pap stains on DNA and RNA integrity and to address the mechanism by which each staining compromises molecular based-analysis. The analyzed studies demonstrate that H&E- and Pap-staining result in low DNA recovery and some degree of DNA fragmentation. Additionally, it is concluded that hemalum inhibits PCR by interfering with DNA extraction, preventing DNA polymerase attachment and possibly by rescuing divalent cations. Accordingly, proper sample purification and adjustment of PCR conditions are of key importance to achieve satisfactory results by PCR in H&E- and Pap-stained samples. Furthermore, although H&E results in RNA fragmentation, it is possible to perform expression analysis in H&E-stained frozen sections, using RNase-free conditions, low amounts of hematoxylin and a rapid protocol from sample collection to RNA analysis. It The effect of Pap-staining on RNA integrity remains to be determined.

The dosage-dependent prenatal caffeine exposure adversely affects levels of integrin αVβ3 and MMP-9 in a rat model of embryo implantation

In the pregnancy period, it is recommended to limit the consumption of caffeine.​ However, the mechanisms of caffeine effect during pregnancy are not fully known.​ In our study, we aimed to investigate the effect of prenatal caffeine consumption on the embryonic implantation in rats as well as shed light on the relationship between the molecules and implantation stages.​MATERIALS AND METHODS: ​Forty-five Wistar albino pregnant rats were randomly divided into 3 main groups, namely into control, low‑dose and high‑dose groups, representing the dose-dependent effects of caffeine. Each main group was divided into 3 subgroups, namely those to be sacrified on days 4 (pre-implantation), 5 (peri-implantation) and 6 (post-implantation) ​. Different doses of caffeine were given on consecutive days, starting from day 1 of pregnancy up to the day of euthanasia. The implantation sites were investigated with the use of hematoxylin et eosin, Masson trichrome and immunostaining of VEGF, MMP-9, integrin αVβ3, mucin-1 and HB-EGF. ​ Prenatal caffeine consumption in rats resulted in a dose-dependent decrease in the number of implantation sites.​ It has been shown that the immunoreactivity of integrin αVβ3 and MMP-9 underwent a change. ​ It has been shown that the levels of integrin αVβ3 and MMP-9 were decreased by prenatal caffeine consumption in rats, which resulted in a decrease in embryo implantation in a dose-dependent manner, especially in the high-dose group (Fig. 5, Ref. 36). Text in PDF www.elis.sk Keywords: caffeine, embryo implantation, integrin, MMP-9.

All-trans-retinoic acid activated mast cells via Mas-related G-protein-coupled receptor-X2 in retinoid dermatitis.

Retinoic acid (RA)-induced dermatitis is the most frequent side-effect limiting its widespread use. However, the exact mechanisms triggering dermatitis are not fully understood, including the role of skin mast cells. The newly discovered Mas-related G-protein-coupled receptor-X2 (MRGPRX2) in mast cells mediates pseudoallergic drug reactions in several types of dermatitis. A possible contribution of MRGPRX2 to contact dermatitis induced by RA has hitherto not been examined. To investigate whether all-trans-RA (ATRA) activates mast cells via MRGPRX2/MrgprB2 (the mouse orthologue), contributing to the pathogenesis of retinoid-induced dermatitis. Wild-type (WT) and MrgprB2-/- mice were treated with topical ATRA to observe local inflammation and mast cell degranulation in vivo by the use of haematoxylin and eosin and immunofluorescence staining. Release of histamine and release of β-hexosaminidase were measured and calcium influx was detected in Laboratory of Allergic Disease 2 (LAD2) cells with specific knockdown targeting MRGPRX2 by small interfering RNA (siRNA) and in primary cells from MrgprB2-/- mice. As compared with WT mice, MrgprB2-/- mice showed resistance to ATRA-triggered contact dermatitis and local inflammatory reactions in the paws. ATRA activated mast cells via the MrgprB2 pathway in murine cells, and via the MRGPRX2 pathway in human mast cells. ATRA-induced dermatitis could be achieved by activating mast cells via MRGPRX2/MrgprB2, which may provide a potential therapy target to reduce the side-effect.

CD8+CD103+ iTregs inhibit the progression of lupus nephritis by attenuating glomerular endothelial cell injury.

We previously reported that ex vivo TGF-β and IL-2-induced CD8+CD103+ regulatory T cells (CD8+CD103+ iTregs) displayed similar immunosuppressive effect and therapeutic function on lupus mice nephritis to that of CD4+Foxp3+ Tregs. In view of the important role of glomerular endothelial cell (GEC) injury in inflammatory processes in SLE, this study aimed to investigate the nature and mechanism of CD8+CD103+ iTregs-mediated amelioration of LN by attenuating GEC injury. Treg cells from patients with SLE and from healthy controls were characterized by flow cytometry analysis. The expression of pro-inflammatory mediators and VEGF were analysed in healthy controls, patients with SLE and MRL/lpr mice by ELISA, western blot, and real-time quantitative RT-PCR (qRT-PCR). Typical lesions of diffuse proliferative LN were observed in MRL/lpr mice through the use of haematoxylin and eosin, Masson, periodic acid-Schiff, periodic acid-Schiff methenamine, transmission electron microscopy and IF microscopy. Angiogenesis was analysed in GECs by cell investigating proliferation, migration, and tube formation. The results revealed that the frequency of Treg cells was inversely correlated with the expression of VCAM-1 and ICAM-1 in patients with SLE. Furthermore, adoptive transfer of CD8+CD103+ iTregs to MRL/lpr mice was associated with decreased levels of autoantibodies and proteinuria, reduced renal pathological lesions, and lowered renal deposition of IgG/C3. We further found that CD8+CD103+ iTregs not only suppressed the expression of pro-inflammatory mediators but also attenuated GEC injury by promoting angiogenesis. Our study has identified the role of CD8+CD103+ iTregs on attenuating GEC injury and provided a possible application of this new iTregs subset in lupus nephritis and other autoimmune diseases.

Decellularized Amniotic Membrane Scaffold as a Pericardial Substitute: An InVivo Study.

In the development of new biomaterials for pericardium substitute, acellular amniotic membrane (AAM) presents potential for new applications in regenerative medicine. We studied an AAM as a pericardial substitute to achieve a suitable, cost effective, abundant matrix for the purpose of using it as graft for tissue repair. Twenty Wistar rats were randomly divided into 2 groups (n= 10/group) and had their pericardiums excised. In the experimental group, the excised pericardium segment was substituted by a 7-mm-diameter patch of decellularized AAM sutured to the lesion area. After 4 weeks, the heart's outer layer of both groups was evaluated. The structure and component characteristics of the scaffold were determined with the use of hematoxylin and eosi, Alizarin Red S, and immumohistochemical staining and scanning electron microscopy. Histopathologic examination of the AAM patches revealed that the integrity of the AAM was preserved, and no calcification was observed on the surface of the myocardium. We also observed thicker pericardium repair tissue in the AAM group compared with the control group. AAM patches, by virtue of their low immunogenicity, evoked minimal host-versus-graft reaction. We conclude that AAM appears to be an ideal substitute for pericardium lesions, because it is integrated into the biologic tissue owing to its low immunogenicity and its ability to diminish the occurrence of adhesions and scarring, increasing the pericardium thickness.

Evaluation of Explanted CorMatrix Intracardiac Patches in Children With Congenital Heart Disease

Animal data demonstrate that intracardiac patches of decellularized porcine small intestine submucosa (CorMatrix; CorMatrix Cardiovascular, Inc, Atlanta, GA) become repopulated with native cells, suggesting the possibility of a substrate for regenerative tissue in humans. We report the only prospective series to date of explanted CorMatrix patches placed in infants with congenital heart disease. CorMatrix patches were implanted as the hemi-Fontan baffle in patients during their second stage of palliation. The patch material was explanted as part of the standard Fontan operation. Specimens were analyzed with the use of hematoxylin and eosin, Movat pentachrome, and trichrome stains. Of the 12 implantations, 10 specimens were explanted. Two patients did not undergo Fontan because of unfavorable hemodynamics. Acellular material,chronic inflammation, fibrosis, and foreign body giant cell reaction was seen in all explanted patches. Also noted in explanted specimens was calcification(n= 2), elastic fibers (n= 2), and eosinophils(n=2).No explanted CorMatrix material showed evidence of ingrowth of native cells or transformation into cardiac tissue at a median of 21 months after implantation. Although the CorMatrix remained pliable and most did not exhibit calcification, these intracardiac patches did not show evidence of native heart tissue ingrowth at a median of 21 months invivo.

Apoptosis in oral epithelial dysplastic lesions and oral squamous cell carcinoma: A prognostic marker.

Apoptotic index (AI) using light microscopy as an indirect measure to assess the significance of apoptosis as a proliferative marker in dysplastic lesions and malignant epithelial lesions of the oral cavity. (1) To quantify the apoptotic bodies/cells in oral epithelial dysplastic (OED) lesions and oral squamous cell carcinoma (OSCC). (2) To measure AI in OED and OSCC. (3) To compare AI in OED and OSCC. The proposed laboratory-based retrospective study involved the use of hematoxylin and eosin (H and E)-stained slides of previously diagnosed OED lesions and OSCC from institutional archives. This study constituted 50 cases, each of H and E-stained slides of previously diagnosed cases of OED and OSCC. AI was calculated as the number of apoptotic bodies/cells expressed as a percentage of the total number of nonapoptotic tumor/dysplastic cells counted in each case. Nonparametric tests such as Kruskal-Wallis test and Mann-Whitney test were used. There was a statistically significant increase in AI from OED to OSCC (P = 0.000). Further studies need to be undertaken to detect and understand the apoptotic mechanisms in the progression from OED to OSCC.

Reassessment of H&E stained clot specimens and immunohistochemistry of phosphorylated Stat5 for histological diagnosis of MDS/MPN

Few studies have comprehensively analysed histopathological findings of bone marrow clots for diagnosis of haematopoietic cell dysplasia. In particular, a limited number of studies have assessed the use of haematoxylin and eosin (H&E) staining, which is generally considered less informative than May-Giemsa staining. In the current study, the utility of bone marrow clot specimens for diagnosis was examined using H&E staining and immunohistochemistry. Patients with myelodysplastic syndromes (MDS) and myelodysplastic/myeloproliferative neoplasm (MDS/MPN), including chronic myelomonocytic leukaemia (CMML), atypical chronic myeloid leukaemia (aCML) lacking Philadelphia chromosome, and juvenile myelomonocytic leukaemia (JMML), were selected for histological evaluation. H&E stained specimens were advantageous for observation of atypical basophilic staining of the cytoplasm and nucleus related to dysplasia. This finding was significantly supported for both MDS and MDS/MPN (p < 0.05 versus May-Giemsa staining); therefore, we concluded that H&E staining could be used for identification of dysplastic cells. In addition, despite the loss of tissue structure, phosphorylated Stat5 immunostaining was sufficiently useful for the observation of myelodysplastic blasts. Thus, clot specimens are useful for diagnosis of haematopoietic dysplasia by pathologists.

Molecular diagnostics in soft tissue sarcomas and gastrointestinal stromal tumors.

Soft tissue sarcomas are rare malignant heterogenous tumors of mesenchymal origin with over fifty subtypes. The use of hematoxylin and eosin stained sections (and immunohistochemistry) in the morphologic assessment of these tumors has been the bane of clinical diagnosis until recently. The last decade has witnessed considerable progress in the understanding and application of molecular techniques in refining the current understanding of soft tissue sarcomas and gastrointestinal stromal tumors beyond the limits of traditional approaches. Indeed, the identification of reciprocal chromosomal translocations and fusion genes in some subsets of sarcomas with potential implications in the pathogenesis, diagnosis and treatment has been revolutionary. The era of molecular targeted therapy presents a platform that continues to drive biomarker discovery and personalized medicine in soft tissue sarcomas and gastrointestinal stromal tumors. In this review, we highlight how the different molecular techniques have enhanced the diagnosis of these tumors with prognostic and therapeutic implications.

Human amniotic epithelial cells suppress relapse of corticosteroid-remitted experimental autoimmune disease

Background aimsMultiple sclerosis (MS) is considered to be a T-cell–mediated disease. Although MS remits with corticosteroid treatment, the disease relapses on discontinuation of therapy. Human amniotic epithelial cells (hAEC) from the placenta are readily accessible in large quantities and have anti-inflammatory properties. Previously we reported that hAEC given near disease onset ameliorated clinical signs and decreased myelin oligodendrocyte glycoprotein (MOG)-specific immune responses in MOG-induced experimental autoimmune encephalomyelitis (EAE), an experimental MS model. MethodsTo examine the therapeutic effect of hAEC in a clinically relevant setting, we first treated MOG peptide–induced EAE mice with a corticosteroid, prednisolone, in drinking water to induce remission. hAEC were then infused intravenously into the remitted mice. Anti-MOG antibodies in serum were detected by enzyme-linked immunoassay. Splenocyte proliferation was assessed by 3H-thymidine incorporation. Immune cell subpopulations in spleens and lymph nodes and secreted cytokines in splenocyte culture were quantified by flow cytometry. Central nervous system histology was examined with the use of hematoxylin and eosin, Luxol fast blue and immunostaining. ResultsWith cessation of prednisolone treatment, hAEC delayed EAE relapse for 7 days, and, after another 7 days, largely remitted disease in six of eight responder mice. Splenocyte proliferation was suppressed, anti-MOG35–55 antibodies in serum were decreased and interleukin-2 and interleukin-5 production by splenocytes were elevated after hAEC treatment. In the central nervous system, hAEC-treated mice had decreased demyelination and fewer macrophages in the inflammatory infiltrates. hAEC treatment also increased CD4+CD25+FoxP3+ regulatory T cells in inguinal lymph nodes. ConclusionsThese data demonstrate that the therapeutic effects of hAEC after corticosteroid treatment in an MS model probably are the consequence of peripheral immunoregulation. We suggest that hAEC may have potential as a cell therapy for remitted MS.

Abstract P2-03-03: Feasibility of evaluation of breast tissue using confocal microscopy with strip mosaicing

Abstract Background: Confocal microscopic strip mosaicing (CSM) provides noninvasive optical sectioning and high resolution, which allows for imaging of nuclear and morphological detail in freshly excised tissue. CSM can image large areas of tissue at micron-level resolution in minutes, which may offer an advantage over standard histology that requires days. We have conducted a preliminary investigation of the feasibility of this technology for the evaluation of breast tissue from surgical excision specimens. Design: In a prospective study, 80 fresh human breast tissue samples from surgical excision specimens of 24 patients were imaged using a prototype confocal strip scanner. Fresh tissue specimens were immersed in Acridine Orange (AO) for 45 seconds to stain the nuclei, then pressed against the glass imaging window and imaged with a 30X, 0.75 numerical aperture (measured) objective lens and a 488 nm laser. Images were acquired in two modes of contrast: in fluorescence (with AO), showing nuclear morphology, and in reflectance (endogenous), showing stroma. The use of fluorescence for nuclear staining mimics the use of hematoxylin in pathology, and the use of reflectance eosin. Use of two contrast modes allows the fluorescence image to be colorized purple and the reflectance image pink, producing confocal strip mosaics that mimic H&amp;E histology in appearance. Specimens were subsequently fixed in formalin and routinely processed to obtain H&amp;E stained sections. H&amp;E and confocal images were compared by the study pathologist (M.M.) Results: Freshly excised breast tissue samples as large as 2 cm x 2 cm were imaged in less than five minutes, with 1-micron resolution and measured optical sectioning of 6 microns. We compared the CSM images against standard histopathology images. In our series we evaluated the following histologies: 12 invasive carcinoma (11 ductal, 1 lobular), 3 ductal carcinoma in-situ, 3 lobular carcinoma in situ, 1 atypical lobular hyperplasia, 1 atypical ductal hyperplasia and various benign lesions such as fat necrosis, fibrocystic changes, and ductal hyperplasia. In confocal images invasive and in situ carcinoma as well as benign ducts and lobules were distinguished from surrounding stromal tissue. Limitations that are typically encountered in standard histology, such as distinguishing low grade ductal carcinoma in situ (DCIS) from lobular carcinoma in situ (LCIS) or atypical proliferations were encountered in the grayscale confocal images as well. Conclusion: In this initial feasibility study, CSM produced images that could be diagnosed as benign or neoplastic by the study pathologist. Further study is needed to build an image library of breast histology and compare reproducibility of histologic diagnoses between CSM (grayscale and colorized images) and traditional optical microscopy, and assess interobserver reproducibility in diagnosis. CSM potentially provides rapid and noninvasive evaluation of breast parenchyma, and has a potential application for intraoperative margin assessment of resected breast specimens. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-03-03.