Conventional Hematoxylin Research Articles

MALDI Imaging for Proteomic Painting of Heterogeneous Tissue Structures.

To present matrix-assisted laser desorption/ionization (MALDI) imaging as a powerful method to highlight various tissue compartments. Formalin-fixed paraffin-embedded (FFPE) tissue of a uterine cervix, a pancreas, a duodenum, a teratoma, and a breast cancer tissue microarray (TMA) are analyzed by MALDI imaging and by immunohistochemistry (IHC). Peptide images are visualized and analyzed using FlexImaging and SCiLS Lab software. Different histological compartments are compared by hierarchical cluster analysis. MALDI imaging highlights tissue compartments comparable to IHC. In cervical tissue, normal epithelium can be discerned from intraepithelial neoplasia. In pancreatic and duodenal tissues, m/z signals from lymph follicles, vessels, duodenal mucosa, normal pancreas, and smooth muscle structures can be visualized. In teratoma, specific m/z signals to discriminate squamous epithelium, sebaceous glands, and soft tissue are detected. Additionally, tumor tissue can be discerned from the surrounding stroma in small tissue cores of TMAs. Proteomic data acquisition of complex tissue compartments in FFPE tissue requires less than 1 h with recent mass spectrometers. The simultaneous characterization of morphological and proteomic features in the same tissue section adds proteomic information for histopathological diagnostics, which relies at present on conventional hematoxylin and eosin staining, histochemical, IHC and molecular methods.

Phosphorylated Histone H3 (PHH3) as a Novel Cell Proliferation Marker and Prognosticator for Meningeal Tumors: A Short Review.

Cell proliferation markers are used in grading of brain tumors and in patient stratification for different treatments. Therefore, defining simple and accurate proliferation markers are of essential importance. Ki67 labeling index is widely studied, yet it is subject to interlaboratory variability. Ki67 protein is expressed on all cell cycle phases except G0 and does not precisely reflect proliferating cells. Counting mitotic figures (MFs) is used in World Health Organization grading of brain tumors (meningiomas), but the detection of MFs on conventional hematoxylin and eosin stained slides is very time consuming. Further, karyorrhexis and pyknosis in apoptotic or crushed cells may mimic mitotic morphology and falsify counting results. Histone H3 phosphorylation on serine-10 is specific to mitosis and phosphorylated histone H3 (PHH3) proliferation markers (as counts defined per area or as indices defined per cell numbers) are increasingly being used to evaluate proliferation in various tumors. Herein, we review current data on PHH3 proliferation markers in meningeal tumors. PHH3-staining highlights mitotic cells and makes easier of rapid grading by driving pathologist's attention on the most mitotically active areas. Thereby, it would function more sensitive in detecting MFs that might be otherwise overloked and more precise by reducing interobserver variability through allowing the pathologist to analyze if the stained nuclei exhibit morphologic features of mitosis.

Hematoxylin and eosin staining of intact tissues via delipidation and ultrasound

Acquiring information on the precise distribution of a tumor is essential to evaluate intratumoral heterogeneity. Conventional hematoxylin and eosin staining, which has been used by pathologists for more than 100 years, is the gold standard of tumor diagnosis. However, it is difficult to stain entire tumor tissues with hematoxylin and eosin and then acquire the three-dimensional distribution of cells in solid tumors due to difficulties in the staining and rinsing. In this paper, we propose a modified hematoxylin and eosin staining method, in which delipidation and ultrasound waves were applied to enhance tissue permeability and accelerate dye diffusion. This improved hematoxylin and eosin staining method is termed iHE (intact tissue hematoxylin and eosin staining). We applied the iHE method to stain intact organs of mice, which were then sectioned and imaged sequentially. The results showed that the whole tissue was stained homogeneously. Combined with micro-optical sectioning tomography (MOST), the iHE method can be used for 3D volume imaging and to evaluate the intratumoral heterogeneity of the entire tumor tissue spatially. Therefore, this method may help to accurately diagnose the invasion stage of tumors and guide clinical treatments.

Molecular profiling of colorectal tumors stratified by the histological tumor-stroma ratio - Increased expression of galectin-1 in tumors with high stromal content.

The tumor microenvironment is a dominant determinant of cancer cell behavior. Reactive tumor stroma is associated with poor outcome perspective. The tumor-stroma ratio (TSR) is a strong independent prognostic factor in colorectal cancer and is easily assessed using conventional hematoxylin and eosin (H&E) stained paraffin sections at the invasive margin of the tumor. We aim to understand the biology of the tumor stroma in colorectal cancer by investigating the transcriptomic profiles of tumors classified by the TSR method. The TSR was assessed in a cohort of 71 colorectal cancer patients undergoing surgery without (neo)adjuvant therapy. In the cohort, stroma-high tumors were distinguished from stroma-low tumors at gene expression level in the upregulation of biological pathways related to extracellular matrix (ECM) remodeling and myogenesis. The activated microenvironment in stroma-high tumors overexpressed different types of collagen genes, THBS2 and 4 as well as INHBA, COX71A and LGALS1/galectin-1. The upregulation of THBS2, COX7A1 and LGALS1/galectin-1. The upregulation of THBS2, COX7A1 and LGALS1/galectin-1 in stroma-high tumors was validated in The Cancer Genome Atlas. In conclusion, the gene expression data reflects the high stromal content of tumors assessed based on the histological method, the TSR. The composition of the microenvironment suggests an altered proteolysis resulting in ECM remodeling and invasive capacity of tumor cells.

Effect of platelet-rich plasma on the skin and follicle development of neonatal mice

Objective To observe the effect of platelet-rich plasma (PRP) on the skin tissue and follicle development of neonatal mice. Methods Forty neonatal C57BL/6 mice were randomly divided into PRP experimental group and normal saline control group, 20 mice in each group. In the middle of the back, single-point intradermal injection of 0.05 ml PRP with blood platelet counts of 109/ml was carried out in the experimental group, and equivalent injection of normal saline was carried out in the control group. The general observation was done on 5 animals each at the 3rd, 5th, 7th and 10th day, and the full thickness skin tissue was cut for paraffin embedding and conventional hematoxylin and eosin (HE) and Masson staining. The histological changes of the tissue sections were observed and photoed. Image J software was used to measure the gray level of skin, and ipp software to measure the thickness of the dermis and the epidermis. Results At the 3rd day, 5th day, and 7th day, the gray values (9 723.30±874.87, 13 371.25±1 511.42, 16 794.70±1 899.03, mean P=0.000) of the skin development color of the mice in PRP group were generally observed. The phenomenon of the changes in the experimental group occurred earlier than that in the control group. The statistical results of the grey values of the skin color were in accordance with the general observation. Simultaneously, at the 10th day, the general observation revealed that the hair density value of the mice in PRP group was significantly higher than that of the mice in the control group. The HE staining results showed that at the 3rd day, 5th day, 7th day, and 10th day, the values of skin tissue epidermal thickness (0.435±0.011, 0.460±0.009, 0.497±0.008, 0.052±0.014; P=0.005, 0.000, 0.000, 0.016) of the mice in the experimental group were all higher than those of the mice in the control group. At the 3rd day, 5th day, 7th day, and 10th day, the values of the skin tissue dermis thickness (0.744±0.011, 1.168±0.016, 1.820±0.011, 1.717±0.012; P=0.000, 0.000, 0.000, 0.034) of the mice in the experimental group were all remarkably higher than those of the mice in the control group during the development of the same number of days. At the 3rd day, 5th day, 7th day, and 10th day, the number of hair follicles (18.33±1.77, 25.33±1.02, 34.33±1.58, 39.33±1.23; P=0.000, 0.000, 0.005, 0.007) of the mice in the experimental group obviously increased as compared with that of the mice in the control group. Furthermore, the hair follicles in the skin tissue of the mice in the experimental group during the development of the same number of days had a better development than that of the mice in the control group. The Masson staining results demonstrated that during the early development of the skin tissue, the new collage secretion volume of the mice in the experimental group was significantly greater than that of the mice in the control group, and the trend of their collagen arrangement to the align of the collagen in mature skin tissue showed earlier in the experimental group than that of the mice in the control group. Conclusion PRP is beneficial for the development and mature of the skin and hair follicles. Key words: Platelet rich plasma; Dermis; Epidermis; Hair follicle; Development; Neonatal mice

Oil Red O Is a Useful Tool to Assess Donor Liver Steatosis on Frozen Sections During Transplantation

Oil Red O is a useful tool to assess donor liver steatosis on frozen sections during transplantation.Steatosis is a frequent finding in liver evaluation during transplantation, accounting for 9% to 26% of biopsied donor liver. The degree of macrovesicular steatosis is classified as mild, moderate, and severe; the latter is considered an absolute contraindication to liver transplantation because it is associated with poor allograft outcome. Because of the scarcity of organs, there is a debate whether livers with less severe macrovesicular steatosis are still suitable for transplant. Consequently, tools or methods that allow a more accurate intraoperative assessment of steatosis on frozen sections are mandatory.The aim of this study is to improve intraoperative evaluation of steatosis during transplantation using Oil Red O stain on liver biopsies. MethodsTwenty consecutive liver biopsies of donors were collected during transplantation procedures from September 2017 to February 2018 at the Institute of Pathology of the University and Hospital Trust of Verona, Italy. Each liver biopsy was cut at a different thickness (3, 5, and 8 μm) and stained with both Oil Red O and conventional hematoxylin and eosin for intraoperative consultation. The degree (percentage of hepatocytes involved) of fatty changes was recorded. The results obtained during the intraoperative consultation were finally compared with the formalin-fixed and paraffin-embedded permanent section. ResultsAssessment of steatosis on hematoxylin and eosin frozen sections was reported as mild in 17 cases (85%), moderate in 2 cases (10%) and severe in 1 case (5%). Oil Red O frozen sections reported the following results: mild steatosis in 16 cases (80%), moderate in 2 cases (10%), and severe in 2 cases (10%). The percentage of liver steatosis obtained with Oil Red O was consistent in all cases with that of the permanent sections. The staining procedure for Oil Red O required approximately 18 minutes. ConclusionsOil Red O special stain is a fast and inexpensive tool to improve the assessment of steatosis on frozen biopsies during liver transplantation.

High-Contrast Imaging of Cholesterol Crystals in Rabbit Arteries Ex Vivo Using LED-Based Polarization Microscopy.

Detection of cholesterol crystals (Chcs) in atherosclerosis disease is important for understanding the pathophysiology of atherosclerosis. Polarization microscopy (PM) has been in use traditionally for detecting Chcs, but they have difficulty in distinguishing Chcs with other crystalline materials in tissue, such as collagens. Thus, most studies using PM have been limited to studying cell-level samples. Although various methods have been proposed to detect Chcs with high specificity, most of them have low signal-to-noise ratios, a high system construction cost, and are difficult to operate due to a complex protocol. To address these problems, we have developed a simple and inexpensive universal serial bus (USB) PM system equipped with a 5700 K cool-white light-emitting diode (LED). In this system, Chcs are shown in a light blue color while collagen is shown in a yellow color. More importantly, the contrast between Chcs and collagens is improved by a factor of 2.3 under an aqueous condition in these PM images. These imaging results are well-matched with the ones acquired with two-photon microscopy (TPM). The system can visualize the features of atherosclerosis that cannot be visualized by the conventional hematoxylin and eosin and oil-red-o staining methods. Thus, we believe that this simple USB PM system can be widely used to identify Chcs in atherosclerosis.

The tumour-stroma ratio in colon cancer: the biological role and its prognostic impact.

The tumour microenvironment consists of a complex mixture of non-neoplastic cells, including fibroblasts, immune cells and endothelial cells embedded in the proteins of the extracellular matrix. The tumour microenvironment plays an active role in tumour behaviour. By interacting with cancer cells, it influences disease progression and the metastatic capacity of the tumour. Tumours with a high amount of stroma correspond to poor patient prognosis. The tumour-stroma ratio (TSR) is a strong independent prognostic tool in colon cancer and provides additional value to the current clinically used tumour-node-metastasis classification. The TSR is assessed on conventional haematoxylin and eosin-stained paraffin sections at the invasive front of the tumour. Here we review studies demonstrating the prognostic significance of the TSR in solid epithelial tumours with a focus on colon cancer. Moreover, the biological role of the tumour microenvironment during tumour progression and invasion will be discussed, as well as the attempts to target the tumour stroma for therapeutic purposes. We suggest that the TSR can be implemented with little effort and without additional costs in current routine pathology diagnostics owing to its simplicity and reliability.

Rapid histopathological imaging of skin and breast cancer surgical specimens using immersion microscopy with ultraviolet surface excitation

Rapid histopathological evaluation of fresh, unfixed human tissue using optical sectioning microscopy would have applications to intraoperative surgical margin assessment. Microscopy with ultraviolet surface excitation (MUSE) is a low-cost optical sectioning technique using ultraviolet illumination which limits fluorescence excitation to the specimen surface. In this paper, we characterize MUSE using high incident angle, water immersion illumination to improve sectioning. Propidium iodide is used as a nuclear stain and eosin yellow as a counterstain. Histologic features of specimens using MUSE, nonlinear microscopy (NLM) and conventional hematoxylin and eosin (H&E) histology were evaluated by pathologists to assess potential application in Mohs surgery for skin cancer and lumpectomy for breast cancer. MUSE images of basal cell carcinoma showed high correspondence with frozen section H&E histology, suggesting that MUSE may be applicable to Mohs surgery. However, correspondence in breast tissue between MUSE and paraffin embedded H&E histology was limited due to the thicker optical sectioning in MUSE, suggesting that further development is needed for breast surgical applications. We further demonstrate that the transverse image resolution of MUSE is limited by the optical sectioning thickness and use co-registered NLM to quantify the improvement in MUSE optical sectioning from high incident angle water immersion illumination.

In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization

Three-dimensional (3D) in vitro microphysiological cultures, such as spheroids and organoids, promise increased patient relevance and therapeutic predictivity compared with reductionist cell monolayers. However, high-throughput characterization techniques for 3D models are currently limited to simplistic live/dead assays. By sectioning and staining in vitro microtissues, researchers can examine their structure; detect DNA, RNA, and protein targets; and visualize them at the level of single cells. The morphological examination and immunochemistry staining for in vitro cultures has historically been done in a laborious manner involving testing one set of cultures at a time. We have developed a technology to rapidly screen spheroid phenotype and protein expression by arranging 66 spheroids in a gel array for paraffin embedding, sectioning, and immunohistochemsitry. The process is quick, mostly automatable, and uses 11 times less reagents than conventional techniques. Here we showcase the capabilities of the technique in an array made up of 11 different cell lines stained in conventional hematoxylin and eosin (H&E) staining, as well as immunohistochemistry staining for estrogen (ER), progesterone (PR), and human epidermal growth factor (Her-2) receptors, and TP53. This new methodology can be used in optimizing stem cell–based models of disease and development, for tissue engineering, safety screening, and efficacy screens in cancer research.

Xylene/Chloroform Free Microwave Tissue Processing and Staining a Non-hazardous and Time Effective Alternative

Introduction: Tissue processing using xylene/chloroform has been employed in histopathology reporting for the past 100 years. Microwave technique has not only reduced the processing time from 1 day to one hour but also doesn’t use xylene/chloroform and has been found to be on par with conventional processing. Xylene is expensive and detrimental to human health. The present study replaces chloroform with isopropanolol in tissue processing and xylene with liquid dish wash solution (LDW) in staining which is not only cheap but also non bio hazardous. Aim: To assess the efficacy of xylene free processing (microwave) and staining versus conventional tissueprocessing and hematoxylin and eosin are staining. Materials and methods: Two Tissue bits from 15 consecutively submitted samples at RL Jalappa Hospital after 1st Jan 2015 each of breast, cervix, lymph node, fat, thyroid, skin, alimentary tract, muscle, salivary gland, liver and kidney were taken and one was processed and stained using conventional method, other using xylene/chloroform free processing and conventional staining and other with both xylene/chloroform free processing and staining. The 495 sections were evaluated and scored by two pathologists independently for nuclear staining, cytoplasmic staining, uniformity, clarity and crispness. Results: In the samples evaluated xylene free processing and staining seems to be on par with conventional xylene processed and stained sections terms of nuclear and cytoplasmic detail, clarity and crispness. However Xylene cleared sections showed slightly better uniform staining (p value->0.05) Conclusion: Xylene/chloroform free processing and staining is not only a rapid but also safe alternative to conventional processing and staining using xylene. However more extensive studies need to be done on other types of tissues for validation

Prognostic Value of Occult Isolated Tumour Cells within Regional Lymph Nodes of Dogs with Malignant Mammary Tumours

Canine mammary tumours (CMTs) are the most common type of neoplasm in bitches. As in women, the presence of metastasis in regional lymph nodes is an important prognostic factor in bitches with mammary carcinomas, but the clinical significance of occult isolated tumour cells (ITCs) within lymph nodes is still undefined in this species. The effectiveness of immunohistochemistry (IHC) in identifying occult ITCs and micrometastasis (MIC) was compared with that of the conventional haematoxylin and eosin staining technique. The relationship between tumour size, histological type, histological grade and the presence of metastasis was evaluated. The overall survival (OS) of female dogs with occult mammary carcinomas and ITCs within lymph nodes was analysed. Fragments of mammary carcinoma and regional lymph nodes of 59 female dogs were also evaluated. Histological sections of mammary carcinoma and lymph node samples were studied for tumour diagnosis and lymph node samples were tested by IHC using a pan-cytokeratin antibody. It was found that 35.2% of occult ITCs and 2.8% of hidden MIC were detected when IHC was used. There was a good correlation between the size of the tumour and metastasis to the lymph nodes (P=0.77). ITCs were observed more frequently in the medullary region (60.7%) and metastases in the cortical region (44.4%). There was no significant difference in the OS between female dogs with occult ITCs and lymph nodes without ITCs. IHC can detect occult tumour cells in lymph nodes that are negative by histopathological examination. Female dogs with nodal ITCs do not have lower survival.

Abstract P2-01-28: Intraoperative assessment of the sentinel node (SN) in breast cancer by one step nucleic acid assay (OSNA): Experience of over 800 patients

Abstract Introduction The intra-operative assessment of the sentinel node in women with breast cancer enables an immediate axillary node clearance if the sentinel node is positive. This has significant benefits for the Patient, the Surgeon and the Health Care Provider. There are a variety of methods for the intra-operative assessment which include: Touch Imprint Cytology, Frozen Section Analysis and Polymerase Chain Reaction (PCR) based molecular assays. OSNA is a molecular assay using PCR, which detects the presence of cytokeratin 19 in the sentinel node. We report our experience of OSNA for the intraoperative assessment of the sentinel node in a single institution with 807 patients. Methods All patients with operable breast cancer who were found to be node negative on clinical and radiological assessment of the axilla, and who had their axilla staged by a sentinel node biopsy at the Breast Unit at Warwick Hospital, UK over a 5 year period were included in this study. Data was collected from a prospective database maintained at the Breast Unit. The axillary node positivity rate and the percentage of patients with macrometastatic and micrometastatic disease as detected by OSNA was collected and compared with a group of 411 patients who had the intraoperative assessment by Touch Imprint Cytology and final histology by conventional Haematoxylin &Eosin(H&E) assessment, prior to the introduction of OSNA. The Chi-square test were used for statistical significance. Results 807 patients had their sentinel assessed intraoperatively using OSNA in this 5 year study period. The sentinel node was positive in 292 patients (36.5%). Of those who had a positive node, 138(17.3%) had macrometastatic disease and 154(19.2%) had micrometastatic disease. When compared to 411 patients in the preOSNA period, that were assessed by Touch Imprint Cytology and H&E sections, the node positivity rate increased from 24.6% to 36.5% (p 0.0001) with the introduction of OSNA. Whilst there was no significant increase in the rate of macrometastatic disease – 21.15% versus 17.3 %( p 0.052), there was a significant increase in the patients who had micrometastases detected on OSNA - 3.5% versus 19.2 %( p 0.0009) as shown in the table. Comparison of OSNA with conventional Touch Imprint Cytology and H&EResult TIC and H&ETIC and H&EOSNAOSNAp value NumberPercentageNumberPercentage PositiveMacro8421.113817.30.052PositiveMicro143.515419.20.0009Negative 31365.451563.5 Conclusion Our results prove that OSNA is a much more sensitive test for picking up metastatic disease, especially micrometastatic disease, in the sentinel node. Whilst this did cause some anxiety in the initial part of the study period, the results of recent trials like ACSOG Z-11 and IBCSG 23-01 have shown that small volume disease or micrometastases in the sentinel node do not always require an axillary node clearance. Intraoperative assessment of the sentinel node with OSNA significantly upstages the axillary nodal status, especially with regard to micrometastatic disease, but the ability to proceed to an axillary node clearance at the same operation as the sentinel node biopsy, still has significant advantages for the Patient, Surgeon and Health Care Providers. Citation Format: Clarke D, Weston S, Ion C, Smith A, Kabir A, Harries S, Jones L. Intraoperative assessment of the sentinel node (SN) in breast cancer by one step nucleic acid assay (OSNA): Experience of over 800 patients [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-01-28.

Peritumoral lymphatic vessel density and invasion detected with immunohistochemical marker D240 is strongly associated with distant metastasis in breast carcinoma

BackgroundDetection of vascular invasion by hematoxylin and eosin staining is the current pathological assessment practice to diagnose breast carcinoma. However, conventional hematoxylin and eosin staining failed to distinguish between blood vessel invasion and lymphatic vessel invasion. Both are important prognostic criteria however with different outcomes. The aim of this study is to distinguish between blood vessel invasion and lymphatic vessel invasion using conventional assessment and immunohistochemical markers. The prognostic significance of both circulatory invasions in invasive breast carcinoma was also investigated.MethodsConsecutive sections of breast carcinoma samples from 58 patients were stained with CD34 and D240 to stain blood and lymphatic vessels respectively. Hematoxylin and eosin staining was carried out on another consecutive section as conventional staining.ResultsAlthough blood vessel density is higher in the sections (median = 10.3 vessels) compared to lymphatic vessel density (median = 0.13), vessel invasion is predominantly lymphatic invasion (69.8 and 55.2% respectively). Interestingly, peritumoral lymphatic vessel density and peritumoral lymphatic invasion was significantly associated with distant metastasis (p = 0.049 and p = 0.05 respectively). The rate of false positive and false negative interpretation by hematoxylin and eosin was 46.7 and 53.3% respectively.ConclusionsLymphatic vessel invasion is a strong prognostic markers of breast carcinoma invasion and the use of immunohistochemical markers increase the rate and accuracy of detection.

Hematoxylin and eosin staining of mucins of the gastrointestinal tract

An infrequent observation of assessing hematoxylin and eosin sections is the blue staining of mucins (for example those in goblet cells). This is believed to be due to a low concentration of alum and high pH of the hematoxylin staining solution. This study examines the incidence of blue mucin in various sites of the gastrointestinal tract using a low alum, high pH hematoxylin solution. The results are compared with a conventional hematoxylin solution, iron alum celestine blue method and an alcian blue (pH 2.5)-periodic acid-Schiff (AB-PAS) stain to characterize the type of mucin demonstrated. This study is the first to offer evidence that blue-stained mucin with low alum, high pH hematoxylin corresponds with carboxylated mucins as shown by the AB-PAS stain in the gastrointestinal tract. Iron alum celestine blue was also found to stain the mucin of a proportion of rectal biopsies and appendix as well as the carboxylated mucin of one duodenal biopsy.

Role of synaptophysin in the intraoperative assessment of quadrantic innervation of the proximal doughnut in Hirschsprung disease.

Symptoms may persist in a retained aganglionic segment of the colon after corrective (pull-through) surgery in Hirschsprung disease (HD). Thus, it is important to assess the proximal doughnut for innervation abnormalities intraoperatively by frozen sections stained with conventional haematoxylin and eosin stain and supported by rapid acetylcholinesterase (AChE) histochemistry. When the doughnut is proximal to the sigmoid colon, AChE is not useful and requires ratification by yet another rapid technique and hence this study. Two pathologists independently evaluated fresh doughnuts from the proximal bowel clinically assumed to be of normal innervation intraoperatively and chosen for anastomosis in patients with HD along with controls using AChE and synaptophysin (SY) immunohistochemistry. From 38 patients with HD, 28 doughnuts (63.7%) showed normal innervation with intense SY activity in the mucosa, the muscularis and the ganglion cells. The circumferential aganglionic doughnuts (abnormal innervation) (n= 6, 13.6%) showed neither SY-positive fibres in the mucosa nor in the muscularis. The abnormal transition zone doughnuts (n=10, 22.7%) showed involvement of three quadrants of the doughnut in one, two quadrants in three and one quadrant in six with decreased SY-positive fibres in the muscularis and scattered ganglion cells with a statistically significant measure of agreement of (κ=0.973) between the two. The pattern, intensity and distribution of SY-positive fibres in the muscularis propria of the doughnut of the proximal bowel chosen intraoperatively for anastomosis in HD can identify sectors with abnormal innervation allowing the surgeon to seek normal innervation status more proximally to avoid complications.

Plasma Cell Cheilitis: A Clinicopathological and Immunohistochemical Study of 13 Cases

BackgroundPlasma cell cheilitis is an unusual benign plasma cell proliferative disease of an unknown etiology that typically presents on the lip.ObjectiveThe aim of this study was to investigate the clinicopathological characteristics of 13 cases of plasma cell cheilitis.MethodsThe present study investigated the clinical manifestations, treatment modalities, and outcome of 13 patients diagnosed with plasma cell cheilitis from 2011 to 2016 at Kangdong Sacred Heart Hospital and Hallym University Sacred Heart Hospital. Biopsy specimens of the all cases were evaluated using conventional hematoxylin and eosin staining with kappa and lambda immunoglobulin light chain immunohistochemistry.ResultsThe age of the patients ranged from 39 to 86 years (mean, 64.7 years), with male predominance. Histopathologically, 61.5% and 38.5% of patients showed band-like and pan dermal plasmacytic infiltrates, respectively. Eosinophilic infiltration was noted in 69.2% of patients. All cases showed both kappa and lambda immunoglobulin light chain reactivities, and kappa predominance was confirmed in 9 patients (69.2%). A majority of the patients was treated with local therapy, such as intralesional steroid injection with topical tacrolimus. Among the 13 patients, plasma cell cheilitis completely resolved, partially resolved, and recurred in 3 (23.1%), 5 (38.5%), and 5 patients (38.5%), respectively.ConclusionPlasma cell cheilitis presented as erosive edematous circumscribed patches or plaques affecting mainly the lower lip of elderly male patients. The majority of histopathology cases showed characteristic plasma cell aggregation on the upper dermis that was immunopositive for immunoglobulin light chain, with kappa predominance.

Intraepithelial Extramammary Paget's Disease of the Vulva: The Clinicopathological Characteristics, Management, and Outcome in a Study of 18 Female Patients.

This study aimed to analyze the clinicopathologic characteristics, management, and outcome of patients with intraepithelial extramammary Paget's disease of the vulva (EMPDV). This retrospective study included medical records and pathological slides of 18 female patients with EMPDV between January 2001 and December 2012. Diagnosis was confirmed using conventional hematoxylin and eosin staining. The median age was 65 years. The distribution of lesions was from labium majus, labium minus, clitoris, and distal vaginal wall. The appearance was erythematous patches, hypopigmentation, and hyperpigmentation. The symptoms were pruritus, erosions, burning sensation, and pain. The mean duration of symptoms was 24 months. Ten patients received local excision, 3 accepted wide local excisions and successful skin grafting, and 5 underwent vulvectomies. Frozen sections showed that 3 and 15 patients had positive and negative margins, respectively. The average length of follow-up was 70 months. During a follow-up period, 3 patients were lost. One patient with positive margins and one with negative margins recurred and the overall recurrence rate was 11.1%. The diagnosis of EMPDV is often delayed but has good prognosis. A complete surgical resection is an effective treatment. Skin grafting may be a good method to promote incision healing.

Protective effects of ursolic acid against hepatotoxicity and endothelial dysfunction in mice with chronic high choline diet consumption

This study was designed to investigate the preventive effect of ursolic acid (UA), a plant-based pentacyclic triterpenoid carboxyl acid, against vascular endothelial damage and liver oxidative injury in the mice fed with 3% dietary high choline (HC) water. Mice fed 3% HC water for 8 weeks significantly displayed liver oxidative stress and vascular endothelial dysfunction (p < 0.01). Furthermore, continuous administration of UA at 400 and 800 mg/kg bw in HC-fed mice could significantly inhibit the HC-induced elevation of serum total cholesterol, total triglyceride, low density lipoprotein-cholesterol, endothelin 1 and thromboxane A2 levels as well as alanine aminotransferase and aspartate aminotransferase activities, while the HC-induced decline of serum high density lipoprotein-cholesterol, endothelial nitric oxide synthase, nitric oxide and prostaglandin I2 levels could be markedly elevated following the treatment (p < 0.05, p < 0.01). UA at 400 and 800 mg/kg bw also increased the hepatic total superoxide dismutase and glutathione peroxidase activities and decreased hepatic malonaldehyde and non-esterified fatty acid levels, relative to HC-treated mice (p < 0.05, p < 0.01). Moreover, the conventional haematoxylin and eosin staining observation of the liver and vascular tissues suggested that UA exerted a significant protective role against HC diet-induced endothelial damage and liver injury in mice. This is the first report showing high intake of dietary choline can induce liver damage and UA has the potential preventive effect against vascular and liver injury in HC-fed mice.

Label-free Raman spectroscopic imaging to extract morphological and chemical information from a formalin-fixed, paraffin-embedded rat colon tissue section.

SummaryAnimal models and archived human biobank tissues are useful resources for research in disease development, diagnostics and therapeutics. For the preservation of microscopic anatomical features and to facilitate long‐term storage, a majority of tissue samples are denatured by the chemical treatments required for fixation, paraffin embedding and subsequent deparaffinization. These aggressive chemical processes are thought to modify the biochemical composition of the sample and potentially compromise reliable spectroscopic examination useful for the diagnosis or biomarking. As a result, spectroscopy is often conducted on fresh/frozen samples. In this study, we provide an extensive characterization of the biochemical signals remaining in processed samples (formalin fixation and paraffin embedding, FFPE) and especially those originating from the anatomical layers of a healthy rat colon. The application of chemometric analytical methods (unsupervised and supervised) was shown to eliminate the need for tissue staining and easily revealed microscopic features consistent with goblet cells and the dense populations of cells within the mucosa, principally via strong nucleic acid signals. We were also able to identify the collagenous submucosa‐ and serosa‐ as well as the muscle‐associated signals from the muscular regions and blood vessels. Applying linear regression analysis to the data, we were able to corroborate this initial assignment of cell and tissue types by confirming the biological origin of each layer by reference to a subset of authentic biomolecular standards. Our results demonstrate the potential of using label‐free Raman microspectroscopy to obtain superior imaging contrast in FFPE sections when compared directly to conventional haematoxylin and eosin (H&E) staining.