Conventional Histopathological Examination Research Articles

Role of Endoscopic Ultrasound-Fine Needle Aspiration in Management of Submucosal Lesions (SMLs) of GIT and Significant Predictors of Malignancy of GIST Lesions

Background: Submucosal lesion is a mass or bulge covered by normal-appearing mucosa identified during standard endoscopy. Endoscopic Ultrasound (EUS) allows precise differentiation of the individual layers of the gastrointestinal (GI) tract. Endoscopic Ultrasound-Fine Needle Aspiration (EUS-FNA) permits safe and accurate sampling of submucosal lesions (SMLs) for further histopathology. Gastrointestinal stromal tumors are the commonest mesenchymal neoplasms of the GI tract. They should be stratified by malignant potential according to tumor size, location, and mitotic count. Objectives: The aim of work was to evaluate the role of endoscopic ultrasound with EUS-FNA in management of submucosal gastrointestinal lesions, with special concern about gastrointestinal stromal tumors (GISTs) lesions and the criteria highly associated with malignant nature. Patients and methods: This multi-center prospective study included a total of 150 patients with gastrointestinal submucosal lesions, conducted to evaluate EUS and EUS-FNA as diagnostic tools of submucosal lesions, including 68 GIST lesions. All patients underwent conventional endoscopy, EUS-FNA and histopathological examination of the samples obtained in Theodor Bilharz Research Institute, Cairo University Hospital and Kobry El Qubba Military Hospital from 2015 to 2017. Results: There were 150 patients with submucosal GI lesions, including 93(62%) males and 57(38%) females, with the mean age of 52±11.9 years. Presentations included dyspepsia 73(48.7%), asymptomatic SMLs 31(20.7%). The lesions were mainly gastric 102(68%). Final diagnoses were GIST 68 (45.3%), lipoma 11(7.3%), leiomyoma 9(6%) and extramural lesions 7(4.7%). There were 68 patients with GIST lesions, including 43(63.2%) males and 25(36.8%) females, with the mean age of 54±10.3 years. Presentations included dyspepsia 41(63.2%), asymptomatic SMLs 16(23.5%). The lesions were mainly gastric 57(83.8%). Sensitivity and specificity were 88.6% and 100% of EUS-FNA as diagnostic tools for GIST lesions respectively. (PPV) and (NPV) were 100% and 82.8% respectively. Conclusion: It could be concluded that EUS and EUS-FNA were highly significant methods in diagnosis of GIST lesions in relation to final histopathology.

Histopathological analysis of the inflammatory response of two invasive techniques in the calcaneal tendon of a mouse

Abstract Background and Aim Percutaneous needle electrolysis (PNE) is a therapeutic tool which has demonstrated its effectiveness of the treatment of tendinopathies and muscle problems. Previous authors have based the therapeutic effect of the same on the ability to provoke an important inflammatory response. However, there is a scarcity of basic research directed at understanding its effects on a cellular and histological level. The aim of this study was to histopathologically compare the inflammatory response provoked by PNE versus dry needling (DN) in a healthy animal model. Material and Methods For the proposed study aim a murine model was used with a total of 18 C57BL/6J mice. All animals received an application of DN in the left common calcaneal tendon, whereas on the right, treatment with PNE was applied (3 impacts of 3mA during 3 seconds). Groups of animals were sacrificed at 3, 7 and 15 days to evaluate the histopathological evolution of the lesions. Samples of the common calcaneal tendon were taken and fixed in commercial formalin at 4% tamponed during 24 hours, processed and included in paraffin blocks. Sections of 3 microns thick were performed which were dyed with hematoxylin-eosin for conventional histopathological examination. Results Dry needling of the common calcaneal tendon induced the presence of an acute inflammatory infiltrate (characterized by the presence of polymorphonuclear neutrophils and macrophages) in the peritenon and adjacent fat tissue which was detectable after the third day, and which was most notable on days 7 and 14. The application of PNE induced a pattern with a similar behavior, however on day 14, the inflammatory infiltrate of PNE was more evident compared with DN. Conclusion The proposed in vivo murine model has been useful for the study of the first phases of the regeneration process induced by both techniques. The histopathological results show that PNE generates a superior stimulus compared to DN on day 14.

Inflammatory response of two invasive techniques in the mouse with collagenase induced tendinopathy

Abstract Background Percutaneous needle electrolysis (PNE) is a physiotherapy technique which has demonstrated its effectiveness for the treatment of tendinopathies and muscle pathologies. All the authors consulted base its therapeutic effect on the capacity to produce an important inflammatory response during the initial phase of the regenerative process. However, presently, the histological effects of the technique on tendinopathies are unknown. The aim of this study was to histologically examine the inflammatory response provoked by the PNE versus dry needling (DN) in mice with collagenase-induced tendinopathy. Material and Methods In order to achieve the proposed aim, a murine model was used with a total of 18 C57BL/6J mice. One week prior to the intervention, collagenase-induced tendinopathy was performed in the common calcaneal tendons of all mice. Once the pathology was established, they received DN in the left common calcaneus tendon, whereas, on the right, they received treatment with PNE. Groups of animals were sacrificed at 3, 7 and 14 days to evaluate the histological evolution of injuries. Samples of the common calcaneal tendon were taken and were later fixed in formalin, processed, and included in paraffin blocks. Sections of 3 microns thick were performed, which were dyed using hematoxylin-eosin for conventional histopathological examination. Results In both groups, tissue damage was observed with collagen fragmentation and the presence of fibroblasts. Dry needling of the common calcaneal tendon induced the presence of an acute inflammatory infiltrate (characterized by the presence of polymorphonuclear neutrophils and macrophages) at the level of the peritenon and adjacent fat tissue which was detectable after day 3, and notable after days 7 and 14. The application of PNE induced a similar effect, although on day 14, the inflammatory infiltrate of PNE was more evident, compared to DN. Conclusion The proposed in vivo murine model has demonstrated to be useful for the study of the evolution of the first phases of the regenerative process induced by both techniques. The histopathological results reveal that PNE generates a pro-inflammatory stimulus which is superior to DN on day 14.

Dynamic observation and analysis of metabolic response to moxibustion stimulation on ethanol-induced gastric mucosal lesions (GML) rats

BackgroundGastric mucosal lesion (GML) is the initiating pathological process in many refractory gastric diseases. And moxibustion is an increasingly popular alternative therapy that prevents and treats diseases. However, there are few published reports about developing pathology of GML and therapeutic mechanism of moxibustion treatment on GML. In this study, we investigated pathology of GML and therapeutic mechanism of moxibustion treatment on GML.MethodsThe male Sprague-Dawley (SD) rats were induced by intragastric administration of 75% ethanol after fasting for 24 h and treated by moxibustion at Zusanli (ST36) and Liangmen (ST21) for 1 day, 4 days or 7 days. Then we applied 1H NMR-based metabolomics to dynamic analysis of metabolic profiles in biological samples (stomach, cerebral cortex and medulla). And the conventional histopathological examinations as well as metabolic pathways assays were also performed.ResultsMoxibustion intervention showed a beneficial effect on GML by modulating comprehensive metabolic alterations caused by GML, including energy metabolism, membrane metabolism, cellular active and neurotransmitters function.ConclusionsMoxibustion can effectively treat gastric mucosal damage and effectively regulate the concentration of some related differential metabolites to maintain the stability of the metabolic pathway.

Effects of sevoflurane on reproductive function of male rats and its main mechanism of action

Objective: To study the effects of sevoflurane on reproductive function and its main mechanism of action in male rats.Materials and methods: Forty adult male Sprague-Dawley rats were divided into 4 groups and exposed to 0, 50, 300 and 1800 ppm of sevoflurane, respectively. After 15 days, the serum levels of sex hormones and inflammatory factors were detected using enzyme-linked immunosorbent assay. Left testis was taken for conventional histopathological examination and TUNEL staining. Right testis was used for sperm production and daily sperm count were evaluated daily. Johnsen score was used to categorize the spermatogenesis. The expression of related genes in the hypothalamic-pituitary-gonadal axis were analyzed by quantitative real time polymerase chain reaction (qRT-PCR).Results: Exposure to sevoflurane increased the levels of serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), tumor necrosis factor-α (TNF-α), and monocyte chemoattractant protein 1 (MCP-1), decreased the content of serum testosterone (T), reduced the concentration of testicular sperm, the production of daily sperm and Johnsen score, and damaged vas deferens in a dose dependent manner. In addition, chronic exposure to sevoflurane down-regulated transcription of gonadotropin-releasing hormone (GnRH) and kisspeptin (Kiss)-1 as well as its receptor GPR54 in hypothalamus, attenuated GnRH receptor and LH-β mRNA levels, but increased FSH-β mRNA in pituitary gland, and enhanced mRNA of LH receptor and FSH receptor, but decreased INH-α and INH-βA mRNA levels in testes.Discussion and conclusions: Sevoflurane induces disorders of spermatogenesis and causes testicular injury. The underlying mechanism may be related to the imbalance of sex hormones in the hypothalamic-pituitary-gonadal axis.

Patent Highlights April–May 2019

Patent Highlights April–May 2019

Dynamic Analysis of Metabolic Response in Gastric Ulcer (GU) Rats with Electroacupuncture Treatment Using 1H NMR-Based Metabolomics.

Gastric ulcer (GU), a common digestive disease, has a high incidence and seriously endangers health of human. According to the previous studies, it has been proved that electroacupuncture at acupoints of stomach meridian had a good effect on GU. However, there are few published studies on metabolic response in gastric ulcer (GU) rats with electroacupuncture treatment. Herein, we observed the metabolic profiles in biological samples (stomach, liver, and kidney) of GU rats with electroacupuncture treatment by 1H NMR metabolomics combined with pathological examination. The male SD rats were induced by intragastric administration of 70% ethanol after fasting for 24 hours and treated by electroacupuncture at Zusanli (ST36) and Liangmen (ST21) for 1 day, 4 days, or 7 days, respectively. And the conventional histopathological examinations as well as metabolic pathways assays were also performed. We found that GU rats were basically cured after electroacupuncture treatment for 4 days and had a complete recovery after electroacupuncture treatment for 7 days by being modulated comprehensive metabolic changes, involved in the function of neurotransmitters, energy metabolism, cells metabolism, antioxidation, tissue repairing, and other metabolic pathways. These findings may be helpful to facilitate the mechanism elucidating of electroacupuncture treatment on GU.

Evolutionary Expression of HER2 Conferred by Chromosome Aneuploidy on Circulating Gastric Cancer Cells Contributes to Developing Targeted and Chemotherapeutic Resistance.

Purpose: Previous human epidermal growth factor receptor-2 (HER2)-derived resistance studies were based on ex vivo models, which could not mirror evolutionary expression of HER2 during therapy. To investigate dynamic expression of HER2 and its contribution to developing therapeutic resistance conferred by chromosome aneuploidy, both the HER2 phenotype and chromosome 8 (Chr 8) aneuploidy on circulating tumor cells (CTC) were coexamined in advanced gastric cancer (AGC) patients.Experimental Design: A total of 115 AGC patients, including 56 of histopathologic HER2+ (hHER2+) subjects who received first-line HER2-targeted therapy plus chemotherapy, and 59 of hHER2- patients who received chemotherapy alone, were prospectively enrolled. Both HER2 phenotype and Chr8 aneuploidy of CTCs in patients were coexamined by HER2-iFISH during therapy.Results: A fluctuated positive HER2 phenotype on CTCs (cHER2+) was revealed, showing cHER2+ at different time intervals during treatment. Acquisition of the cHER2+ phenotype in 91.0% of hHER2+ and 76.2% hHER2- patients was demonstrated to correlate with development of resistance to trastuzumab-targeted therapy for hHER2+ patients and chemotherapy alone for hHER2- patients. Aneuploid Chr8 was demonstrated to participate in the acquisition of the cHER2+ phenotype, which provides a growth advantage to HER2+ CTCs against therapeutic pressure, leading to the development of therapeutic resistance.Conclusions: Compared with low positivity of conventional histopathologic hHER2 examination routinely performed once, significant higher positivity of cHER2+ on CTCs was observed. Continuously examining cHER2 shows unique advantages with respect to monitoring therapeutic resistance in real time in carcinoma patients. Moreover, contribution of chromosome aneuploidy to the phenotypic evolution of HER2 expression on CTCs may help elucidate underlying mechanisms of developing therapeutic resistance. Clin Cancer Res; 24(21); 5261-71. ©2018 AACR.

Robotic lateral oropharyngectomy following diagnostic tonsillectomy is oncologically safe in patients with high risk human papillomavirus related squamous cell cancer.

Diagnostic tonsillectomy is rarely an oncologic operation owing to close or positive margins. The standard of care is for further treatment to the primary site, typically with adjuvant radiotherapy. 14 patients with close or positive margins following a diagnostic tonsillectomy underwent transoral robotic surgery (TORS) and lateral oropharyngectomy; five patients with the longest follow-up had their excision specimens examined with a step serial sectioning technique (SSS). Conventional histopathological examination of the TORS resection specimens did not demonstrate residual carcinoma in 13 patients, confirmed by examination using SSS in 5 patients. There were no post-operative complications or long-term functional deficit. Seven patients received surgery alone with 100% overall and disease specific survival, respectively (median follow-up 27.5months; range 5.2-50.4). This prospective study suggests that TORS lateral oropharyngectomy alone is an oncologically safe treatment when close or positive margins are identified on diagnostic tonsillectomy in HPV-positive SCC.

Fluorescence Detection of Cell Death in Liver of Mice Treated with Thioacetamide

The purpose of this study was to detect cell death in the liver of mice treated with thioacetamide (TAA) using fluorescence bioimaging and compare this outcome with that using conventional histopathological examination. At 6 weeks of age, 24 mice were randomly divided into three groups: group 1 (G1), control group; group 2 (G2), fluorescence probe control group; group 3 (G3), TAA-treated group. G3 mice were treated with TAA. Twenty-two hours after TAA treatment, G2 and G3 mice were treated with Annexin-Vivo 750. Fluorescence in vivo bioimaging was performed by fluorescence molecular tomography at two hours after Annexin-Vivo 750 treatment, and fluorescence ex vivo bioimaging of the liver was performed. Liver damage was validated by histopathological examination. In vivo bioimaging showed that the fluorescence intensity was increased in the right upper part of G3 mice compared with that in G2 mice, whereas G1 mice showed no signal. Additionally ex vivo bioimaging showed that the fluorescence intensity was significantly increased in the livers of G3 mice compared with those in G1 or G2 mice (p < 0.05). Histopathological examination of the liver showed no cell death in G1 and G2 mice. However, in G3 mice, there was destruction of hepatocytes and increased cell death. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining confirmed many cell death features in the liver of G3 mice, whereas no pathological findings were observed in the liver of G1 and G2 mice. Taken together, fluorescence bioimaging in this study showed the detection of cell death and made it possible to quantify the level of cell death in male mice. The outcome was correlated with conventional biomedical examination. As it was difficult to differentiate histological location by fluorescent bioimaging, it is necessary to develop specific fluorescent dyes for monitoring hepatic disease progression and to exploit new bioimaging techniques without dye-labeling.

Occurrence of splenic leiomyosarcoma in dog: anatomopathological findings

Leiomyosarcoma is a malignant neoplasm with origin in smooth muscles. It can originate in the vessel wall and interstitium of the spleen smooth muscle, which is considered rare. The objective of this study was to describe clinical and anatomopathological findings of a splenic leiomyosarcoma in an eight-year-old bitch of the Fila breed. This splenic leiomyosarcoma weighed 9.2 kg, and had 30×27×14 cm in dimension, rounded shape, irregular outline, reddish color alternating with pale areas, friable and hemorrhagic tissues, and necrotic center. After conventional histopathological examination, the suspected diagnosis (leiomyosarcoma) was confirmed by histochemical analysis using Masson's trichrome staining (which showed the reactivity of the muscle tissue) and by immunohistochemical analysis, showed positivity for smooth muscle actin, vimentin, and desmin. This study emphasizes the importance of associating histopathological evaluation with histochemical and immunohistochemical techniques for a diagnosis of this neoplasm. The occurrence of leiomyosarcoma reported here denotes the importance of considering it for a differential diagnosis in cases of splenic tumors.

Ex Vivo Confocal Fluorescence Microscopy for Rapid Evaluation of Tissues in Surgical Pathology Practice.

- Optical imaging techniques are currently available for imaging tissues without the need for any type of extensive tissue preparation. There are several applications for their potential use in surgical pathology practice. - To evaluate the feasibility of using a confocal fluorescence microscopy (CFM) platform for ex vivo examination of tissues obtained from surgical resections of breast, lung, kidney, and liver. - Tissue fragments (0.5-1.0 cm) were immersed in 0.6 mM acridine orange for 6 seconds and imaged using a CFM platform at a 488-nm wavelength. The imaged tissues were subsequently fixed in formalin and processed routinely to generate hematoxylin-eosin-stained tissue sections. Mosaics of the grayscale CFM images were studied at different magnifications for recognition of the tissue and were compared with conventional histopathologic examination of hematoxylin-eosin tissue sections. - We imaged 55 tissue fragments obtained from 16 breast (29%), 18 lung (33%), 14 kidney (25%), and 7 liver (13%) surgical excision specimens. Acridine orange labeled the nuclei, creating the contrast between nucleus and cytoplasm and thereby recapitulating the tissue architecture. We could obtain CFM images of good quality within 5 to 10 minutes that allowed recognition of the cytomorphologic details for categorization of the imaged tissue and were similar to histologic examination of hematoxylin-eosin tissue sections. - The ease and speed of acquisition of CFM images together with the resolution and resemblance of the CFM images to hematoxylin-eosin sections suggest that the CFM platform has excellent potential for use in surgical pathology practice.

ADENOID CYSTIC CARCINOMA EXHIBITING EXCLUSIVELY SOLID VARIANT: CASE REPORT

Adenoid Cystic Carcinoma (ACC) is an uncommon lesion of the salivary glands composed of modified myoepithelial and ductal cells. Conventional histopathologic examination is the main tool for diagnosis, although in some cases immunohistochemical analysis is necessary. The present report describes a case of ACC with unusual histopathologic appearance and immunohistochemical markers used as auxiliary tool for the diagnosis. A 67-year-old female patient displaying nodular and erythematous lesion localized in the left ventral surface of the tongue. On suspicion of epidermoid carcinoma, incisional biopsy was performed. The histopathologic examination showed small nests of cells with squamous malignant characteristics exhibiting exclusively solid pattern with muscular and neural invasion. The immunohistochemical test proved positive for the following markers p53, 34β12, CK5/6, EMA, CK14, S-100 and c-Kit, which are characteristic of ACC. The patient underwent hemiglossectomy and supraomohyoid left empty. Currently, the patient is being observed and there is no recurrence of the lesion.

Case report of a ganglioneuroma with metaplasia by adipocytes: use of a multi-disciplinary team as a key aid in difficult diagnosis and combined surgery

Herein, we present a rare case of a dumb-bell ganglioneuroma (GN) with metaplasia by adipocytes that was surgically resected in a 24-year-old woman who was asymptomatic. Because of the difficult diagnosis with no consensus about the location of the tumor, diagnostic and surgical procedures were performed with the assistance of a multi-disciplinary team (MDT) and the use of video-assisted thoracoscopic surgery (VATS). After conventional thoracotomy, histopathological and immunohistochemical examinations revealed that the tumor consisted of spindle cells, ganglion cells, and adipose cells. The histopathological diagnosis was GN with metaplasia by adipocytes. The tumor was resected, and the patient had no postoperative complications during a 3-year follow-up. For tumors whose location cannot be confirmed in the posterior mediastinum or in the retroperitoneum, VATS is an effective confirmatory tool. The recruitment of a MDT can be invaluable for formulating and providing treatment options for the patient.

Case report of a ganglioneuroma with metaplasia by adipocytes: use of a multi-disciplinary team as a key aid in difficult diagnosis and combined surgery

: Herein, we present a rare case of a dumb-bell ganglioneuroma (GN) with metaplasia by adipocytes that was surgically resected in a 24-year-old woman who was asymptomatic. Because of the difficult diagnosis with no consensus about the location of the tumor, diagnostic and surgical procedures were performed with the assistance of a multi-disciplinary team (MDT) and the use of video-assisted thoracoscopic surgery (VATS). After conventional thoracotomy, histopathological and immunohistochemical examinations revealed that the tumor consisted of spindle cells, ganglion cells, and adipose cells. The histopathological diagnosis was GN with metaplasia by adipocytes. The tumor was resected, and the patient had no postoperative complications during a 3-year follow-up. For tumors whose location cannot be confirmed in the posterior mediastinum or in the retroperitoneum, VATS is an effective confirmatory tool. The recruitment of a MDT can be invaluable for formulating and providing treatment options for the patient.

Diagnosis of periprosthetic joint infection using alpha-defensin test or multiplex-PCR: ideal diagnostic test still not found.

Diagnosing periprosthetic infection remains a challenge. Multiplex-PCR and biomarkers such as alpha-defensin are potentially useful and fast methods for detecting periprosthetic infection. This study compared these new methods with clinical assessment, conventional microbiological methods and histo-pathological examination. Twenty-eight consecutive patients with 30 joints and a mean age of 67.7years (range 39 to 88) with removal of total hip arthroplasty (THA) or total knee replacement (TKR) were included in this study. Patients were classified according to the modified Musculoskeletal Infection Society score (MSIS) for infected joints. Punction fluid and tissue specimens were taken for conventional microbiological examination, alphadefensin test was performed, a synovial membrane specimen was used for multiplex-PCR and histopathological examination was carried out. The alpha-defensin test and multiplex-PCR showed a sensitivity of 76.9 vs. 30.8% and a specificity of 82.4 vs. 100%, respectively. We found a significant difference between the positive and negative results (p = 0.0023). The conventional microbiological methods were not significantly different from the alpha-defensin test (p = 0.244) with a sensitivity of 84.6% and a specificity of 100% but did differ significantly from the multiplex PCR (p = 0.0030). There was a significant difference between modified MSIS classification and multiplex PCR (p = 0.0007). Neither alpha-defensin test nor multiplex-PCR could detect periprosthetic infection immediately and reliably. Multiplex-PCR was suitable for detecting the non-infected but not the truly infected. Alpha-defensin test was helpful but showed no satisfactory results. Conventional microbiological methods remain the most reliable for periprosthetic infection diagnosis.

Ex vivo sentinel lymph node mapping in colorectal cancer using a magnetic nanoparticle tracer to improve staging accuracy: a pilot study.

Nodal status is the most important prognostic factor in colorectal cancer (CRC). Small occult metastases may remain undetected on conventional histopathological examination, potentially resulting in undertreatment. Ex vivo sentinel lymph node mapping (SLNM) can be used to improve the accuracy of nodal staging, but the currently used tracers suffer from drawbacks, which hamper implementation of the technique in routine clinical practice. Magnetic tracers are the optimal size for sentinel lymph node (SLN) retention and allow objective quantitative selection of SLNs; they therefore have great potential for SLNM in CRC. The study evaluates the feasibility of exvivo magnetic SLNM and compares the performance of this technique with blue dye SLNM. Twenty-eight exvivo SLNM procedures were performed in 27 histological node-negative patients with CRC using a magnetic tracer and blue dye. A magnetometer was used to select magnetic SLNs after formalin fixation of the CRC specimen. Both magnetic and blue SLNs were subjected to serial sectioning andimmunohistochemical staining to reveal occult metastases. At least one SLN was successfully identified in 27/28 (96%) and 25/28 (89%) of the cases with the magnetic technique and blue dye. Isolated tumour cells were detected in 10 patients. This was predicted with 100% sensitivity and accuracy using the magnetic technique, and with 91% sensitivity and 96% accuracy using the blue dye technique. This study demonstrates that exvivo magnetic SLNM is a feasible technique for use in routine clinical practice, improving nodal staging accuracy of CRC patients.

Quantitative Metabolomic Profiling of Plasma, Urine, and Liver Extracts by 1H NMR Spectroscopy Characterizes Different Stages of Atherosclerosis in Hamsters.

Atherosclerosis (AS) is a progressive disease that contributes to cardiovascular disease and shows a complex etiology, including genetic and environmental factors. To understand systemic metabolic changes and to identify potential biomarkers correlated with the occurrence and perpetuation of diet-induced AS, we applied 1H NMR-based metabolomics to detect the time-related metabolic profiles of plasma, urine, and liver extracts from male hamsters fed a high fat and high cholesterol (HFHC) diet. Conventional biochemical assays and histopathological examinations as well as protein expression analyses were performed to provide complementary information. We found that diet treatment caused obvious aortic lesions, lipid accumulation, and inflammatory infiltration in hamsters. Downregulation of proteins related to cholesterol metabolism, including hepatic SREBP2, LDL-R, CYP7A1, SR-BI, HMGCR, LCAT, and SOAT1 was detected, which elucidated the perturbation of cholesterol homeostasis during the HFHC diet challenge. Using "targeted analysis", we quantified 40 plasma, 80 urine, and 60 liver hydrophilic extract metabolites. Multivariate analyses of the identified metabolites elucidated sophisticated metabolic disturbances in multiple matrices, including energy homeostasis, intestinal microbiota functions, inflammation, and oxidative stress coupled with the metabolisms of cholesterol, fatty acids, saccharides, choline, amino acids, and nucleotides. For the first time, our results demonstrate a time-dependent metabolic progression of multiple biological matrices in hamsters from physiological status to early AS and further to late-stage AS, demonstrating that 1H NMR-based metabolomics is a reliable tool for early diagnosis and monitoring of the process of AS.

Fluorescence In Situ Hybridization for Melanoma Diagnosis: A Review and a Reappraisal.

Although conventional histopathological examination is the undisputable mainstay for the diagnosis of melanocytic skin neoplasms, fluorescence in situ hybridization (FISH) has the potential to provide important information to morphologically challenging cases. The standard melanoma FISH test targeting RREB1 (6p25), MYB (6q23), CCND1 (11q13), and centromere 6 is an effective compromise between cost, technical complexity, and sensitivity. The authors use the standard FISH-positivity as a tie-breaker for challenging melanocytic neoplasms mainly in a non-Spitzoid morphologic context because the currently available test leaves several unresolved issues: namely, a relatively low diagnostic accuracy in morphologically ambiguous melanocytic neoplasms; a relatively low sensitivity and specificity in Spitzoid neoplasms; and the occurrence of false positives due to tetraploidy in Spitz nevi and in nevi with an atypical epithelioid component. Under investigation is currently a new melanoma probe cocktail targeting RREB1 (6p25), C-MYC (8q24), CDKN2A (9p21), and CCND1 (11q13). However, CDKN2A is a significant parameter only if lost in homozygosis, and this complicates the interpretation of the results. Furthermore, the new melanoma probe cocktail has been tested on cases of atypical Spitzoid proliferations with fatal outcomes which at present are too few to allow definite conclusions. The authors propose the implementation of a FISH algorithm (standard 4-probe test followed by either C-MYC or CDKN2A/centromere 9) to assist the histopathological diagnosis and minimize the technical problems. Nevertheless, because the diagnostic accuracy of the FISH technique is far from being absolute, the overall clinicopathological context must always guide the decision-making process about the management of morphobiologically ambiguous melanocytic proliferations.

In vivo real-time assessment of colorectal polyp histology using an optical biopsy forceps system based on laser-induced fluorescence spectroscopy.

In order to reduce time, costs, and risks associated with resection of diminutive colorectal polyps, the American Society for Gastrointestinal Endoscopy (ASGE) recently proposed performance thresholds that new technologies should meet for the accurate real-time assessment of histology of colorectal polyps. In this study, we prospectively assessed whether laser-induced fluorescence spectroscopy (LIFS), using the new WavSTAT4 optical biopsy system, can meet the ASGE criteria. 27 patients undergoing screening or surveillance colonoscopy were included. The histology of 137 diminutive colorectal polyps was predicted in real time using LIFS and findings were compared with the results of conventional histopathological examination. The accuracy of predicting polyp histology with WavSTAT4 was assessed according to the ASGE criteria. The overall accuracy of LIFS using WavSTAT4 for predicting polyp histology was 84.7 % with sensitivity, specificity, and negative predictive value (NPV) of 81.8 %, 85.2 %, and 96.1 %. When only distal colorectal diminutive polyps were considered, the NPV for excluding adenomatous histology increased to 100 % (accuracy 82.4 %, sensitivity 100 %, specificity 80.6 %). On-site, LIFS correctly predicted the recommended surveillance intervals with an accuracy of 88.9 % (24/27 patients) when compared with histology-based United States guideline recommendations; in the 3 patients for whom LIFS- and histopathology-based recommended surveillance intervals differed, LIFS predicted shorter surveillance intervals. From the data of this pilot study, LIFS using the WavSTAT4 system appears accurate enough to allow distal colorectal polyps to be left in place and nearly reaches the threshold to "resect and discard" them without pathologic assessment. WavSTAT4 therefore has the potential to reduce costs and risks associated with the removal of diminutive colorectal polyps.