Search

Based on DNA‐methylation, ependymomas growing in the spinal cord comprise two major molecular types termed spinal (SP‐EPN) and myxopapillary ependymomas (MPE(‐A/B)), which differ with respect to their clinical features and prognosis. Due to the existing discrepancy between histomorphogical diagnoses and classification using methylation data, we asked whether deep neural networks can predict the DNA methylation class of spinal cord ependymomas from hematoxylin and eosin stained whole‐slide images. Using explainable AI, we further aimed to prospectively improve the consistency of histology‐based diagnoses with DNA methylation profiling by identifying and quantifying distinct morphological patterns of these molecular ependymoma types. We assembled a case series of 139 molecularly characterized spinal cord ependymomas (nMPE = 84, nSP‐EPN = 55). Self‐supervised and weakly‐supervised neural networks were used for classification. We employed attention analysis and supervised machine‐learning methods for the discovery and quantification of morphological features and their correlation to the diagnoses of experienced neuropathologists. Our best performing model predicted the DNA methylation class with 98% test accuracy and used self‐supervised learning to outperform pretrained encoder‐networks (86% test accuracy). In contrast, the diagnoses of neuropathologists matched the DNA methylation class in only 83% of cases. Domain‐adaptation techniques improved model generalization to an external validation cohort by up to 22%. Statistically significant morphological features were identified per molecular type and quantitatively correlated to human diagnoses. The approach was extended to recently defined subtypes of myxopapillary ependymomas (MPE‐(A/B), 80% test accuracy). In summary, we demonstrated the accurate prediction of the DNA methylation class of spinal cord ependymomas (SP‐EPN, MPE(‐A/B)) using hematoxylin and eosin stained whole‐slide images. Our approach may prospectively serve as a supplementary resource for integrated diagnostics and may even help to establish a standardized, high‐quality level of histology‐based diagnostics across institutions—in particular in low‐income countries, where expensive DNA‐methylation analyses may not be readily available. [ABSTRACT FROM AUTHOR]

Background: Hematoxylin & Eosin (H & E) stains have been conventionally used to establish the status of safe margins following resection of primary Oral Squamous Cell Carcinoma. Due to non-specificity of this stain, there is a possibility of false negative results. In this study, we have assessed the role of Immunohistochemistry (IHC) in establishing the status of safe margins. Aim: To compare Hematoxylin & Eosin (H & E) and Immunohistochemistry (IHC) staining in identification of tumor cells in establishing the status of safe margins. Methodology: This study included 14 cases diagnosed with OSCC. Following resection, the primary lesion was subjected to Histopathological analysis. 2 sets of HP slides were prepared from serial sectioning of the wax block prepared for each of the four margins. Both sets of slides were stained with H &E stain. One set of these slides was further stained with Pan CK marker (IHC) which is a cytokeratin marker to identify tumour cells. Results: All the slides with H & E staining reported negative for tumor infiltration and 4 slides (3 patients) out of 56 were reported positive with PanCK marker. There was a statistically significant difference in the number of patients with positive margins using IHC as compared to H & E stain. Conclusion: Immunohistochemistry using PanCK marker proved to be more efficient in the determination of status of safe margins than routine H & E staining. [ABSTRACT FROM AUTHOR]

Background Enhancing graft fat survival remains a paramount challenge in autologous fat transplantation surgeries. Dedifferentiated fat cells (DFATs) and adipose-derived stem cells (ASCs) represent 2 pivotal cells with potential to improve fat graft survival rates. Objectives In this study we aimed to compare the effectiveness of DFATs and ASCs in promoting fat graft survival, emphasizing their adipogenic and angiogenic capabilities. Methods Both in vitro and in vivo experiments were conducted. In vitro assessments compared adipogenesis, angiogenesis, osteogenesis, chondrogenesis, cell migration abilities, and surface markers. For in vivo evaluation, a cell-assisted lipotransfer animal model was employed to gauge graft volume retention and histological morphology. Analysis techniques included hematoxylin and eosin staining, Western blotting, and real-time polymerase chain reaction. Results In vitro findings suggested a slight superiority of DFATs in adipogenesis and angiogenesis compared to ASCs. In vivo tests demonstrated both cell types surpassed the control in terms of graft volume retention, with the DFATs group marginally outperforming in retention rates and the ASC group presenting a slightly enhanced graft tissue structure. Conclusions Our study underscores the distinct advantages of DFATs and ASCs in bolstering fat graft survival, offering potentially novel insights for plastic surgeons aiming to elevate fat graft survival rates. [ABSTRACT FROM AUTHOR]

Gingival Stillman's cleft is one of the least-studied mucogingival defects that may jeopardize the periodontal health and esthetic of the affected teeth. The etiology behind this lesion is believed to be multifactorial, and the histopathology remains unclear. In this report, we present a case of composite gingival Stillman's cleft in anterior maxillary teeth that was clinically treated with a laterally moved coronally advanced flap. The cleft tissue was removed during root coverage surgery and then was harvested for histopathological analysis using hematoxylin and eosin, Masson's trichrome, and van Gieson's stain. In the cleft site, microscopic examination revealed variable degrees of epithelial bifurcations with elongated forking of rete ridges into the stroma. Endothelial-lined blood vessels and inflammatory cells, primarily lymphocytes and fibroblasts, were seen in the stroma. The Masson trichrome (blue) and Van Gieson (pink) revealed colored gingival tissue with prominent collagen fiber distribution at the cleft site, which is suggestive of gingival fibrous hyperplasia brought on by repeated damage from tooth brushing.

Gingival Stillman's cleft is one of the least-studied mucogingival defects that may jeopardize the periodontal health and esthetic of the affected teeth. The etiology behind this lesion is believed to be multifactorial, and the histopathology remains unclear. In this report, we present a case of composite gingival Stillman's cleft in anterior maxillary teeth that was clinically treated with a laterally moved coronally advanced flap. The cleft tissue was removed during root coverage surgery and then was harvested for histopathological analysis using hematoxylin and eosin, Masson's trichrome, and van Gieson's stain. In the cleft site, microscopic examination revealed variable degrees of epithelial bifurcations with elongated forking of rete ridges into the stroma. Endothelial-lined blood vessels and inflammatory cells, primarily lymphocytes and fibroblasts, were seen in the stroma. The Masson trichrome (blue) and Van Gieson (pink) revealed colored gingival tissue with prominent collagen fiber distribution at the cleft site, which is suggestive of gingival fibrous hyperplasia brought on by repeated damage from tooth brushing. [ABSTRACT FROM AUTHOR]

Background: Keratin is an intermediate protein that structures the cytoskeleton of all epithelial cells. Keratin pearl is present in various pathological conditions such as oral squamous cell carcinoma (OSCC). Mitotic figures can be used as a valuable tool in assessing cellular proliferation and act as a prognostic indicator in dysplasia and malignancy of the oral cavity. Aim: The aim of this study was to compare the efficacy of Leishman's stain with hematoxylin and eosin (H and E) stain in assessing keratin pearls and mitotic figures. Materials and Methods: In total, 30 diagnosed cases of OSCC of three different histological grades were taken. In each case, two sections of 4–5-micron thickness were made and stained with routine H and E stain and Leishman's stain. Staining intensity and specificity results were analyzed statistically using the Chi-square test and P = 0.05 was considered to be statistically significant. Results: Both stains can distinguish three different types of OSCC histologically. Leishman's stain demonstrated staining intensity (P = 0.087) and specificity (P = 0.017) comparable to that of routine H and E. Conclusion: Leishman's stain distinctly stained the keratin pearl and mitotic figures with a higher intensity. However, H and E showed a higher specificity.

Until now, studies on nail diseases have been performed through microscopic diagnosis and microscopic computed tomography (micro-CT). However, these kinds of conventional methods have some limitations. Firstly, the microscopic method is considered the gold standard for medical diagnosis. However, due to the use of fluorescent materials, the sample is damaged and it takes a long time to get results. Secondly, while micro-CT is a noninvasive method to get inner structure images of the sample with high resolution, the penetration and spatial resolution are insufficient for studying the microstructures of the sample, such as the sponge bone and the muscle fibers. In contrast, synchrotron radiation (SR) X-ray imaging technology has the advantage of very vividly demonstrating the anatomic structure of the sample with high penetration, sensitivity and resolution. In this study, we compared the optical microscopic method using hematoxylin and eosin staining and SR imaging to analyze the nail tissue in a mouse model. The results showed that SR could depict the inner structures of a mouse nail without any physical damage. Additionally, we could divide the important anatomical structures of the nail unit into three parts with three-dimensional (3D) images: the nail bed, nail matrix and hyponychium. The images showed that SR could be used for analyzing nails by visualizing the relatively clear and medically semantic structures in a 3D section. We expect that the results of this study will be applied to study nail diseases and conduct pharmaceutical research on their treatment. [ABSTRACT FROM AUTHOR]

A whole-slide imaging (WSI) system is a digital color imaging system used in digital pathology with the potential to substitute the conventional light microscope. A WSI system digitalizes a glass slide by converting the optical image to digital data with a scanner and then converting the digital data back to the optical image with a display. During the digital-to-optical or optical-to-digital conversion, a color space is required to define the mapping between the digital domain and the optical domain so that the numerical data of each color pixel can be interpreted meaningfully. Unfortunately, many current WSI products do not specify the designated color space clearly, which leaves the user using the universally default color space, sRGB. sRGB is a legacy color space that has a limited color gamut, which is known to be unable to reproduce all color shades present in histology slides. In this work, experiments were conducted to quantitatively investigate the limitation of the sRGB color space used in WSI systems. Eight hematoxylin and eosin (H and E)-stained tissue samples, including human bladder, brain, breast, colon, kidney, liver, lung, and uterus, were measured with a multispectral imaging system to obtain the true colors at the pixel level. The measured color truth of each pixel was converted into the standard CIELAB color space to test whether it was within the color gamut of the sRGB color space. Experiment results show that all the eight images have a portion of pixels outside the sRGB color gamut. In the worst-case scenario, the bladder sample, about 35% of the image exceeded the sRGB color gamut. The results suggest that the sRGB color space is inadequate for WSI scanners to encode H and E-stained whole-slide images, and an sRGB display may have insufficient color gamut for displaying H and E-stained histology images. [ABSTRACT FROM AUTHOR]

Background: Diagnosis of initial epithelial pathology maybe difficult in Squamous Cell Carcinoma (SCC), Carcinoma In Situ and other atypical epithelial malignancies, under routine Haematoxylin and Eosin (H and E) stain. The detection of minor basement membrane alterations in doubtful cases is both time consuming and confusing. Aims: To evaluate efficacy of Modified Cajal's Trichrome Stain (CTS) in relation to Haematoxylin and Eosin for study of epithelial dysplasia, carcinoma in situ, micro invasive SCC, frank SCC, and SCC in lymph nodes. Materials and Methods: Formalin-fixed, paraffin-embedded tissue blocks of mild epithelial dysplasia (n = 2), moderate epithelial dysplasia (n = 2), severe epithelial dysplasia (n = 4), carcinoma in situ (n = 1), micro-invasive SCC (n = 4), verrucous carcinoma (n = 1), and frank OSCC (n = 5) were stained with CTS and H&E. The sections were compared based on set histopathological criteria. Results and Conclusion: In SCC cases stained with CTS, invasion into connective tissue and keratin pearls were strikingly evident. Depth of invasion could be more accurately determined. Tumour cells in lymph node were intensely contrasted and easily discernible. Thus, CTS is a good differential stain, clearly delineating the epithelial elements from the connective tissue elements visually. This helps in tracing the basement membrane very clearly. It is an economic, rapid and easy to use method which cannot replace Haematoxylin and Eosin stain in cancer diagnosis, but can definitely be used adjunctive to it. Prompt diagnosis is crucial to effective treatment, and this stain assists in early and rapid diagnosis of cancer.

In this study, we demonstrated for the first time the neuroprotective role of edaravone (Eda) (5 and 10 mg/kg b.w.), a potent free radical scavenger against the unilateral stereotaxic induction of quinolinic acid (QA) (300 nm/4 μl saline)-induced Huntington disease (HD)-like symptoms in behavioral, biochemical, and histological features in male Wistar rats striatum. QA induction, which mimics the early stage of HD, commonly causes oxidative stress to the cell and decreases the antioxidant defense mechanism by altering the level of lipid peroxidation (LPO), protein carbonyls, and nitrate concentration (NO) and the activities of glutathione family enzymes (GPx, GST, GR) and acetyl choline esterase concentration (AChE) which was found to be ameliorated by Eda treatment in both the tested doses 5 and 10 mg/kg b.w. in the significance of P < 0.05 and P < 0.01, respectively. Finally histopathological analysis by hematoxylin and eosin stain concluded the promising neurodefensive role of Eda in rat striatum at the dosage of 10 mg/kg b.w., with the decreased tissue damage and the number of damaged granular cells when compared to QA-induced groups. [ABSTRACT FROM AUTHOR]

Background: 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. Aims: (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. Settings and Design: 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. Materials and Methods: 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. Statistical Analysis Used: Nonparametric tests such as Kruskal–Wallis test and Mann–Whitney test were used. Results: There was a statistically significant increase in AI from OED to OSCC (P = 0.000). Conclusions: Further studies need to be undertaken to detect and understand the apoptotic mechanisms in the progression from OED to OSCC.

Background: Micronucleus (MN) represents small, additional nuclei formed by the exclusion of chromosome fragments or whole chromosomes lagging at mitosis. MN rates, therefore, indirectly reflect chromosome breakage or impairment of the mitotic apparatus. During the last few decades, micronuclei ("MNi") in oral exfoliated epithelial cells are widely used as biomarkers of chromosomal damage, genome instability and cancer risk in humans. However, until now only little attention has been given to the effect of different staining procedures on the results of these MN assays. Aim: To compare the MNi frequencies in oral exfoliated epithelial cells using three different stains, i.e.,Feulgen stain, Papanicolaou stain (Pap) and hemotoxylin and eosin stain (H and E). Materials and Methods: Oral exfoliated cells from 45 cases of potentially malignant disorders (15 oral submucous fibrosis, 15 lichen planus and 15 leukoplakia) and 15 controls with healthy mucosa, were taken and MNi frequencies (No. of MNi/1000 cells) were compared using three different stains. Results: Mean MNi frequency in cases was found to be 3.8 with Feulgen stain, 16.8 with PAP and 25.9 with H and E. In controls, mean MNi frequency was 1.6 with Feulgen stain, 7.7 with PAP and 9.6 with H and E stain. Statistically significant value (P < 0.01) were observed when the three stains were compared together using Kruskal Walli′s ANOVA test. Conclusion: Feulgen being a DNA-specific stain gave the least counts, although statistically significant results from the comparison of MNi frequency between cases and controls were obtained with all the three stains.

Background: Various types of benign and malignant splenic tumours including hemangiomas, lymphagiomas, hamartomas, hemangiosarcomas, malignant lymphomas and metastatic carcinomas share radiological characteristics making it impossible for a physician to determine the definite aetiology of splenic masses noninvasively without histopathological evaluation. It is important that physicians recognize the importance of a careful and continuous follow-up since inflammatory pseudotumours (IPT) are considered to be tumours with an intermediate malignant potential based on their behaviour when they arise in other locations. Methods: Our patient, a 60-year-old woman was evaluated using laboratory, gastroscopy, computed tomography and surgical methods. The resected tumour was evaluated using immunohistochemical methods. Results: The patient presented with weight loss, nausea and vomiting, symptoms lasting over a course of a few months. The splenic mass was found incidentally at the time of work up for gastritis and cholelithiasis. Histologically, the tumour differed from typical splenic architecture being composed of atypical spindle cells with inflammatory elements; numerous plasma cells, macrophages, eosinophils and lymphocytes. Conclusions: Although very difficult to diagnose at initial presentation, it is very important for the physician to be aware of the importance of a careful diagnosis since IPT of the spleen are rare and considered to be tumours with an intermediate malignant potential [ABSTRACT FROM AUTHOR]

Background: Helicobacter pylori is the causative organism for chronic active gastritis, duodenal ulcer and also for malignancies like gastric adenocarcinoma and mucosa associated lymphoid tissue lymphoma. It is essential to mention the presence of H. pylori in gastric biopsies as it has an important role in patient care. Though there are several special stains to detect H. pylori in histological sections, their specificity and sensitivity vary greatly. Immunohistochemically H. pylori can be detected by using anti H. pylori antibody, which reacts with somatic antigens of the whole bacteria. The aim of this study was to compare the reliability of routine hematoxylin and eosin (H and E), Giemsa, Warthin-Starry (WS) silver stain and immunohistochemical technique in diagnosing H. pylori. Materials and Methods: In this retrospective 1-year (2009) study, endoscopic gastric biopsies taken from patients during gastrointestinal-endoscopy with histopathological diagnosis of gastritis were studied. Standard H and E staining was performed on 5-μm-sections from paraffin block of each specimen. Microscopic sections of biopsy specimens of patients showing features of gastritis histopathologically in routine H and E stain and where the presence of H. pylori was suspected were also stained with Giemsa, WS, and immunohistochemistry (IHC) using purified polyclonal H. pylori antiserum (BioGenex). We have not included gastric resection specimens in our study. Results: Of the 29 cases, 26 (32.9%) showed presence of H. pylori on H and E, Giemsa and WS stains, whereas 49 (62.0%) cases demonstrated H. pylori on IHC stain. Conclusion: We conclude that H. pylori detection by IHC has advantage over routine H and E staining. However, in the developing countries with financial constraints, routine H and E staining in combination with special staining are fairly reliable in demonstrating H. pylori. [ABSTRACT FROM AUTHOR]

Aim: The aim of the present study was to stain the known keratin containing tissues by Haematoxylin and eosin stain (H-E), ayoub-shklar (A/S) and modified Papanicolaou (PAP) stain and to compare the efficacy of modified PAP staining procedure with that of A/S stain and H-E staining technique, so as to device a staining procedure which is easy and effective for keratin. Materials and Methods: A Total Number of 60 paraffin embedded tissue sections of known keratin containing tissues including normal keratinized oral mucosa (NKOM), Keratinized Odontogenic Keratocyst (OKC), Verrucous Carcinoma (VC) and Well differentiated Squamous Cell Carcinoma (WDSCC) were taken and 3 sections of 4 microns thickness of each block were cut and stained with above mentioned three stains. Results: Surface keratin was stained distinctly and uniformly in all the three staining techniques in NKOM, OKC, and VC and WDSCC. But results were statistically significant in WDSCC when Amount of keratin pearls and Pattern of staining were compared in all 3 staining procedures and p value was p=0.000 and p=0.001 respectively. Conclusion: Based on the above findings we conclude that the efficacy of modified PAP is comparable with that of H-E stain and A-S stain for surface keratin and thus be used effectively to stain Surface keratin. Whereas to know the exact pattern of cytokeratin expression in SCC a more sensitive tool like immunohistochemical method can be applied. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate the effects of a GENESIS-BCP™ scaffold composed of hydroxyapatite (HAp) and β-tricalcium phosphate ( β-TCP) on bone formation. The relative ratio of components forming GENESIS-BCPTM was investigated by confirming the molar ratio of Ca/P through energy dispersive spectrometer (EDS) analysis. The results being that the scaffold is comprised of 60% HAp and 40% β-TCP. Moreover, scanning electron microscopy (SEM) images showed a unique surface morphology similar to a soccer ball. X-ray diffraction (XRD) analysis showed that the porosity, crystallinity, and Ca/P ratio were 65.5%, 84.9%, and 1.61%, respectively. In vitro tests including cytotoxicity and cell proliferation showed a good biocompatibility of GENESIS-BCP™. In vivo tests, including mouse skull test, and hematoxylin and eosin stains of GENESIS-BCP™ implanted in rabbit and human subjects showed that the bone graft is effective to accelerate new bone formation. It was concluded that GENESIS-BCP™ can be used as an effective alloplast bone graft. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Background: Perls' stain is routinely used to demonstrate iron in liver biopsies. We tested the hypothesis that it may be unnecessary in cases, where no iron or another similar pigment was seen on the routine hematoxylin and eosin (H and E) stained section. Aim:The aim of this study was to evaluate the efficiency of H and E stain in demonstrating iron in liver biopsies as well as to determine the possibility of replacing Perls' stain with H and E stain. Materials and Methods: Two hundred pairs of slides of liver biopsies were taken from the archival files of the Department of Pathology from 2006 to 2011. Perls' and H and E slides were independently reviewed for the presence of iron. Results: Hundred and one cases showed the presence of iron using H and E stain. 84 of 86 cases showed positive iron using both Perls' and H and E stains. Seventeen cases were positive using H and E stain but negative with Perls'. Only two cases did not show the presence of iron using H and E stain. Ninety seven cases were negative using both Perls' and H and E stains.H and E stain showed a sensitivity,specificity,accuracy,positive predictive valve, and negative predictive value of 97.67%, 85.08%, 90.5%, 83.16%, and 97.98%, respectively. Conclusion: We demonstrate that the H and E stain is a sensitive method to detect iron pigment in liver biopsies, particularly when present in large quantities. A negative H and E stain might obviate the need for extra Perls' staining, thus saving costs and shortening report turn-around times. [ABSTRACT FROM AUTHOR]

This article describes a patient with intense pain originating from a benign tumor on her leg. After 7 years of enduring this painful condition with no correct diagnosis, the patient finally underwent a biopsy and excision, which revealed the growth was an angioleiomyoma. [ABSTRACT FROM AUTHOR]

A whole-slide imaging (WSI) system is a digital color imaging system used in digital pathology with the potential to substitute the conventional light microscope. A WSI system digitalizes a glass slide by converting the optical image to digital data with a scanner and then converting the digital data back to the optical image with a display. During the digital-to-optical or optical-to-digital conversion, a color space is required to define the mapping between the digital domain and the optical domain so that the numerical data of each color pixel can be interpreted meaningfully. Unfortunately, many current WSI products do not specify the designated color space clearly, which leaves the user using the universally default color space, sRGB. sRGB is a legacy color space that has a limited color gamut, which is known to be unable to reproduce all color shades present in histology slides. In this work, experiments were conducted to quantitatively investigate the limitation of the sRGB color space used in WSI systems. Eight hematoxylin and eosin (H and E)-stained tissue samples, including human bladder, brain, breast, colon, kidney, liver, lung, and uterus, were measured with a multispectral imaging system to obtain the true colors at the pixel level. The measured color truth of each pixel was converted into the standard CIELAB color space to test whether it was within the color gamut of the sRGB color space. Experiment results show that all the eight images have a portion of pixels outside the sRGB color gamut. In the worst-case scenario, the bladder sample, about 35% of the image exceeded the sRGB color gamut. The results suggest that the sRGB color space is inadequate for WSI scanners to encode H and E-stained whole-slide images, and an sRGB display may have insufficient color gamut for displaying H and E-stained histology images., (Copyright: © 2020 Journal of Pathology Informatics.)

Laparoscopic cholecystectomies represent one of the most common procedures of General Surgery. Dropped gallstones during laparoscopic cholecystectomy is a rare occurrence but can potentially lead to morbid complications. We present a rare case of a broncho-pleuro-cutaneous fistula in a patient with recurrent abscess formations after an elective laparoscopic cholecystectomy. The fistula formed over 3 years from a gallstone that eroded through the diaphragm, into the pleural space, and eventually into a terminal bronchiole. Bilirubin staining within the fistula tract was confirmed on hematoxylin and eosin staining. [ABSTRACT FROM AUTHOR]

Introduction: Histological stains are dyes that bind to a variety of tissues. Modified Gallego's (MG) stain is a modification of Lille's stain that can be used as a differential stain for identification of hard tissues in oral pathological lesions. Objectives: The objective of this study was to identify the presence of hard tissues such as enamel, dentin and cementum in normal extracted teeth and odontogenic tumors using MG stain and to compare the efficacy of MG stain with hematoxylin and eosin (H&E) stain. Methods: A total of fifty samples, twenty decalcified sections of teeth and thirty cases of odontogenic tumors, were included in the present study. Two sections were cut from the above cases and stained with H&E stain and MG stain, respectively, and assessed for the nature of hard tissue. Results: In H&E staining, enamel, dentine, cementum and bone stained pink. Whereas, in MG stain, enamel stained pink, dentin and bone stained green, while cementum stained red. The shade of color differs with the degree of mineralization of the hard tissues in MG stain. Conclusion: MG stain can be used as a differential stain for different hard-tissue structures when compared to routine H and E staining. [ABSTRACT FROM AUTHOR]

Introduction: Histological stains are dyes that bind to a variety of tissues. Modified Gallego's (MG) stain is a modification of Lille's stain that can be used as a differential stain for identification of hard tissues in oral pathological lesions., Objectives: The objective of this study was to identify the presence of hard tissues such as enamel, dentin and cementum in normal extracted teeth and odontogenic tumors using MG stain and to compare the efficacy of MG stain with hematoxylin and eosin (H&E) stain., Methods: A total of fifty samples, twenty decalcified sections of teeth and thirty cases of odontogenic tumors, were included in the present study. Two sections were cut from the above cases and stained with H&E stain and MG stain, respectively, and assessed for the nature of hard tissue., Results: In H&E staining, enamel, dentine, cementum and bone stained pink. Whereas, in MG stain, enamel stained pink, dentin and bone stained green, while cementum stained red. The shade of color differs with the degree of mineralization of the hard tissues in MG stain., Conclusion: MG stain can be used as a differential stain for different hard-tissue structures when compared to routine H and E staining., Competing Interests: There are no conflicts of interest.

Somatic mutations in patient tumor DNA samples can be readily detected based on mass spectrometry. The MassARRAY system is a high-throughput matrix-assisted laser desorption time-of-flight (MALDI) mass spectrometer for detection of nucleic acids. The technique is based on single-nucleotide base extension. A series of PCR assays amplify specific DNA regions of interest harboring mutations. A third primer is then introduced into the reaction which corresponds to the DNA template immediately in front of the mutation site. A final round of PCR is then performed using mass-modified nucleotides. These nucleotides are designed so that no additional bases can be added to the extension primer (terminating bases) after a single-base extension and are mass modified to exaggerate mass differences between nucleotides allowing easier identification by mass spectrometry.The sequences of the extension primer and possible extension products (wild type and mutations) are known; therefore, it is possible to calculate their mass. The mass spectrometer can identify the mass peaks for each assay and identify those with mutations (multiple peaks). The technique was originally designed to screen multiple single-nucleotide polymorphisms (SNPs) in a large number of specimens. A SNP in the coding region of DNA that alters the gene and subsequent protein expression is considered a mutation. Mutations often occur in genes whose protein product is in a key signaling pathway and/or drug target. Rationale treatment options can be designed based upon the presence or absence of these mutations. In this chapter, we describe the process for detection of somatic mutations in DNA extracted from formalin-fixed paraffin-embedded (FFPE) material.

Background: Barr body (BB) evaluation using special stains in various cells has now become a widely accepted technique for human gender identification, particularly for mass screening of the victims of various disasters. Among the various special stains used, recently Aceto Orcein (AO) is suggested to be a better tool in BB identification. Aims: To evaluate 1) the efficiency of AO and hematoxylin and eosin (H&E) staining in identifying BB in pulpal cells and buccal mucosal cells. 2) Reliability of using buccal mucosal cells and pulpal cells in identifying gender. Materials and Methods: The sample size of the study was 30 (15 males and 15 females). Buccal scrapings and pulp tissue taken from the extracted tooth of these subjects were stained with AO and H&E. Two observers blinded with respect to gender of the samples gave the scores. Samples detected with more than five BBs were identified as females. The study was unblindfolded, readings were tabulated and submitted for statistical analysis. Results: The number of BBs identified in AO stained buccal cells ranged from 8-18 for females and 0-6 for males while when H&E was used it ranged from 6-14 for females and 0-5 for males. In pulpal cells, the range was 5-14 for females and 1-5 for males when stained with AO and 7-15 for females and 0-4 for males when stained with H&E. The sensitivity and specificity of AO and H&E for detecting sex accurately in buccal cells was 96% and 93%, respectively. In pulpal cells, it was 67% and 73%. However, usage of AO for detecting BBs was statistically found to be more significant (sig: 0.004) than H&E, when the cells employed were grouped together as a single variable. Conclusion: * AO was found to be more efficient in identifying BBs when buccal cells were used, while in pulpal cells H&E was more efficient. * Among the two type of cells employed here, identification of BB was easier in buccal cells which may be due to adequate cellularity. * Use of pulpal cells for BB detection though difficult and statistically less significant, highlights the possibility for its application in forensic grounds. [ABSTRACT FROM AUTHOR]

Oral delivery of camptothecin has a treatment advantage but is limited by low bioavailability and gastrointestinal toxicity. Poly(amido amine) or PAMAM dendrimers have shown promise as intestinal penetration enhancers, drug solubilizers and drug carriers for oral delivery in vitro and in situ. There have been very limited studies in vivo to evaluate PAMAM dendrimers for oral drug delivery. In this study, camptothecin (5 mg/kg) was formulated and co-delivered with cationic, amine-terminated PAMAM dendrimer generation 4.0 (G4.0) (100 and 300 mg/kg) and anionic, carboxylate-terminated PAMAM generation 3.5 (G3.5) (300 and 1000 mg/kg) in CD-1 mice. Camptothecin associated to a higher extent with G4.0 than G3.5 in the formulation, attributed to an electrostatic interaction on the surface of G4.0. Both PAMAM G4.0 and G3.5 increased camptothecin solubilization in simulated gastric fluid and caused a 2-3 fold increase in oral absorption of camptothecin when delivered at 2 h. PAMAM G4.0 and G3.5 did not increase mannitol transport suggesting that the oral absorption of camptothecin was not due to tight junction modulation. Histologic observations of the epithelial layer of small intestinal segments of the gastrointestinal tract (GIT) at 4 h post dosing supported no evidence of toxicity at the evaluated doses of PAMAM dendrimers. This study demonstrates that both cationic (G.4) and anionic (G3.5) PAMAM dendrimers were effective in enhancing the oral absorption of camptothecin. Results suggest that drug inclusion in PAMAM interior controlled solubilization in simulated gastric and intestinal fluids, and increased oral bioavailability., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Hyperuricemia is related to a variety of pathologies, including chronic kidney disease (CKD). However, the pathophysiological mechanisms underlying disease development are not yet fully elucidated. Here, we studied the effect of hyperuricemia on tryptophan metabolism and the potential role herein of two important uric acid efflux transporters, multidrug resistance protein 4 (MRP4) and breast cancer resistance protein (BCRP). Hyperuricemia was induced in mice by treatment with the uricase inhibitor oxonic acid, confirmed by the presence of urate crystals in the urine of treated animals. A transport assay, using membrane vesicles of cells overexpressing the transporters, revealed that uric acid inhibited substrate-specific transport by BCRP at clinically relevant concentrations (calculated IC50 value: 365±13μM), as was previously reported for MRP4. Moreover, we identified kynurenic acid as a novel substrate for MRP4 and BCRP. This finding was corroborated by increased plasma levels of kynurenic acid observed in Mrp4(-/-) (107±19nM; P=0.145) and Bcrp(-/-) mice (133±10nM; P=0.0007) compared to wild type animals (71±11nM). Hyperuricemia was associated with >1.5 fold increase in plasma kynurenine levels in all strains. Moreover, hyperuricemia led to elevated plasma kynurenic acid levels (128±13nM, P=0.005) in wild type mice but did not further increase kynurenic acid levels in knockout mice. Based on our results, we postulate that elevated uric acid levels hamper MRP4 and BCRP functioning, thereby promoting the retention of other potentially toxic substrates, including kynurenic acid, which could contribute to the development of CKD., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Aging is an inevitable process associated with immune imbalance, which is characterized by a progressive functional decline in major organs, including lung. However, effects of altered Th1/Th2 commitment on lung senescence are largely unknown. To examine effects of altered Th1/Th2 balance on lung aging, we measured proportions of Th1 and Th2 cells and expression of cytokines, chemokines, collagen deposition and other relevant physiological and pathological parameters in 2- and 20-months-old (mo) CXCR3-deficient (CXCR3(-/-)) C57BL/6J mice compared with wild-type (WT) mice. There was a significant weight-loss observed in 20-mo CXCR3(-/-) mice compared with the same aged WT group. Although lung function and structure changed with age in both groups, central airway resistance (Rn), tissue elastance (H) and damping (G) were significantly lower in 20-mo CXCR3(-/-) mice than those of WT mice. In contrast, the whole lung volume (V(L)), the mean linear intercept length of alveolar (L(m)), and the total lung collagen content were significantly elevated in 20-mo CXCR3(-/-) mice. With aging, the lungs of WT mice had typical Th1-type status (increased population of Th1 cells and concentrations of cytokine IFN-γ and CXCR3 ligands) while CXCR3(-/-) mice showed Th2-type polarization (decreased proportion of Th1 cells and concentrations of CXCR3 ligands but increased level of IL-4). Our data suggest that Immunosenescence is associated with lung aging, and that altered Th1/Th2 imbalance favors Th2 predominance in CXCR3(-/-) mice, which contributes to the process of accelerated lung aging in this model., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background: Micronucleus (MN) represents small, additional nuclei formed by the exclusion of chromosome fragments or whole chromosomes lagging at mitosis. MN rates, therefore, indirectly reflect chromosome breakage or impairment of the mitotic apparatus. During the last few decades, micronuclei ("MNi") in oral exfoliated epithelial cells are widely used as biomarkers of chromosomal damage, genome instability and cancer risk in humans. However, until now only little attention has been given to the effect of different staining procedures on the results of these MN assays., Aim: To compare the MNi frequencies in oral exfoliated epithelial cells using three different stains, i.e.,Feulgen stain, Papanicolaou stain (Pap) and hemotoxylin and eosin stain (H and E)., Materials and Methods: Oral exfoliated cells from 45 cases of potentially malignant disorders (15 oral submucous fibrosis, 15 lichen planus and 15 leukoplakia) and 15 controls with healthy mucosa, were taken and MNi frequencies (No. of MNi/1000 cells) were compared using three different stains., Results: Mean MNi frequency in cases was found to be 3.8 with Feulgen stain, 16.8 with PAP and 25.9 with H and E. In controls, mean MNi frequency was 1.6 with Feulgen stain, 7.7 with PAP and 9.6 with H and E stain. Statistically significant value (P < 0.01) were observed when the three stains were compared together using Kruskal Walli's ANOVA test., Conclusion: Feulgen being a DNA-specific stain gave the least counts, although statistically significant results from the comparison of MNi frequency between cases and controls were obtained with all the three stains.