Conventional Histopathological Examination Research Articles

Intraoperative Assessment of Margin Accuracy in Patients Diagnosed With Oral Squamous Cell Carcinoma in a Tertiary Hospital of Jharkhand: Frozen Section Versus Histopathology.

Background The incidence of head and neck carcinoma is increasing. The use of an intraoperative frozen section plays a vital role in the evaluation of margin status in patients undergoing surgery for oral squamous cell carcinoma. A negative margin is not only an indication of successful surgery but also decreases the recurrence of disease and improves the overall survival of patients. Aims and objective The aim of this study is to assess the accuracy of margin in patients undergoing surgery for oral squamous cell carcinoma by intraoperative frozen section and compare it with conventional histopathological examination. Methodology The approach of our study was a hospital-based prospective study conducted on 28 patients diagnosed with oral squamous cell carcinoma. A frozen section was done on all patients undergoing surgery and compared with histopathological examination. Results Out of 28 patients undergoing surgery, the incidence of males was more than females, with a ratio of 6:1. The most common site of the tumor was left buccal mucosa comprising 28.57%, followed by gingivobuccal sulcus comprising 17.85%. In our study, the frozen assessed margin showed a sensitivity of 58.33%, specificity of 98.76%, and accuracy of 95.25%. Conclusion Frozen section is a reliable method for confirmation of margin accuracy and thus reduces the chance of re-surgery and recurrence of disease and increases overall patient survival.

Enhancing accessibility for improved diagnosis with modified EfficientNetV2-S and cyclic learning rate strategy in women with disabilities and breast cancer.

Breast cancer, a prevalent cancer among women worldwide, necessitates precise and prompt detection for successful treatment. While conventional histopathological examination is the benchmark, it is a lengthy process and prone to variations among different observers. Employing machine learning to automate the diagnosis of breast cancer presents a viable option, striving to improve both precision and speed. Previous studies have primarily focused on applying various machine learning and deep learning models for the classification of breast cancer images. These methodologies leverage convolutional neural networks (CNNs) and other advanced algorithms to differentiate between benign and malignant tumors from histopathological images. Current models, despite their potential, encounter obstacles related to generalizability, computational performance, and managing datasets with imbalances. Additionally, a significant number of these models do not possess the requisite transparency and interpretability, which are vital for medical diagnostic purposes. To address these limitations, our study introduces an advanced machine learning model based on EfficientNetV2. This model incorporates state-of-the-art techniques in image processing and neural network architecture, aiming to improve accuracy, efficiency, and robustness in classification. We employed the EfficientNetV2 model, fine-tuned for the specific task of breast cancer image classification. Our model underwent rigorous training and validation using the BreakHis dataset, which includes diverse histopathological images. Advanced data preprocessing, augmentation techniques, and a cyclical learning rate strategy were implemented to enhance model performance. The introduced model exhibited remarkable efficacy, attaining an accuracy rate of 99.68%, balanced precision and recall as indicated by a significant F1 score, and a considerable Cohen's Kappa value. These indicators highlight the model's proficiency in correctly categorizing histopathological images, surpassing current techniques in reliability and effectiveness. The research emphasizes improved accessibility, catering to individuals with disabilities and the elderly. By enhancing visual representation and interpretability, the proposed approach aims to make strides in inclusive medical image interpretation, ensuring equitable access to diagnostic information.

Application of Targeted Coronary Angiography in the Diagnosis of Sudden Cardiac Death.

To diagnose coronary artery stenosis by using the postmortem computed tomography angiography (PMCTA), and to explore the diagnostic value of PMCTA in sudden cardiac death. Six death cases were selected, and the contrast medium iohexol was injected under high pressure through femoral artery approach with 5F pigtail catheter to obtain coronary image data and then the data was analyzed. The results of targeted coronary imaging and coronary artery calcium score (CaS) were compared with the results of conventional autopsy and histopathological examination. The autopsy and histopathological examination of cases with coronary artery stenosis obtained similar results in targeted coronary angiography, with a diagnostic concordance rate of 83.3%. Targeted coronary angiography could effectively show coronary artery diseases, and the CaS was consistent with the results of conventional autopsy and histopathological examination. Targeted coronary angiography can be used as an effective auxiliary method for conventional autopsy in cases of sudden cardiac death.

Three-dimensional analysis of perineural invasion in extrahepatic cholangiocarcinoma using tissue clearing.

Perineural invasion (PNI) is a characteristic invasion pattern of distal cholangiocarcinoma (DCC). Conventional histopathologic examination is a challenging approach to analyze the spatial relationship between cancer and neural tissue in full-thickness bile duct specimens. Therefore, we used a tissue clearing method to examine PNI in DCC with three-dimensional (3D) structural analysis. The immunolabeling-enabled 3D imaging of solvent-cleared organs method was performed to examine 20 DCC specimens from five patients and 8 non-neoplastic bile duct specimens from two controls. The bile duct epithelium and neural tissue were labeled with CK19 and S100 antibodies, respectively. Two-dimensional hematoxylin/eosin staining revealed only PNI around thick nerve fibers in the deep layer of the bile duct, whereas PNI was not identified in the superficial layer. 3D analysis revealed that the parts of DCC closer to the mucosa exhibited more nerves than the normal bile duct. The nerve fibers were continuously branched and connected with thick nerve fibers in the deep layer of the bile duct. DCC formed a tubular structure invading from the epithelium and extending around thin nerve fibers in the superficial layer. DCC exhibited continuous infiltration around the thick nerve fibers in the deep layer. This is the first study using a tissue clearing method to examine the PNI of DCC, providing new insights into the underlying mechanisms.

Quantitative Phase Imaging Using Digital Holographic Microscopy to Assess the Degree of Intestinal Inflammation in Patients with Ulcerative Colitis.

Ulcerative colitis (UC) is characterized by chronic inflammation of the colorectum. Histological remission has emerged as a potential future treatment goal; however, the histopathological assessment of intestinal inflammation in UC remains challenging with a multitude of available scoring systems and the need for a pathologist with expertise in inflammatory bowel disease (IBD). In previous studies, quantitative phase imaging (QPI) including digital holographic microscopy (DHM) was successfully applied as an objective method for stain-free quantification of the degree of inflammation in tissue sections. Here, we evaluated the application of DHM for the quantitative assessment of histopathological inflammation in patients with UC. In our study, endoscopically obtained colonic and rectal mucosal biopsy samples from 21 patients with UC were analyzed by capturing DHM-based QPI images that were subsequently evaluated using the subepithelial refractive index (RI). The retrieved RI data were correlated with established histological scoring systems including the Nancy index (NI) as well as with endoscopic and clinical findings. As a primary endpoint, we found a significant correlation between the DHM-based retrieved RI and the NI (R2 = 0.251, p < 0.001). Furthermore, RI values correlated with the Mayo endoscopic subscore (MES; R2 = 0.176, p < 0.001). An area under the receiver operating characteristics (ROC) curve of 0.820 confirms the subepithelial RI as a reliable parameter to distinguish biopsies with histologically active UC from biopsies without evidence of active disease as determined by conventional histopathological examination. An RI higher than 1.3488 was found to be the most sensitive and specific cut-off value to identify histologically active UC (sensitivity of 84% and specificity of 72%). In conclusion, our data demonstrate DHM to be a reliable tool for the quantitative assessment of mucosal inflammation in patients with UC.

Diagnostic aid to subepidermal calcified nodule with dermoscopy and reflectance confocal microscopy: a case report

BackgroundSubepidermal calcified nodule (SCN) is a type of calcinosis cutis that usually occurs in children. The lesions in the SCN resemble those of other skin diseases, such as pilomatrixoma, molluscum contagiosum, and juvenile xanthogranuloma, leading to a high rate of misdiagnoses. Noninvasive in vivo imaging techniques, represented by dermoscopy and reflectance confocal microscopy (RCM), have dramatically accelerated skin cancer research over the past decade, and their applications have greatly expanded into other skin disorders. However, the features of an SCN in dermoscopy and RCM have yet to be reported previously. Combining these novel approaches with conventional histopathological examinations is a promising method for increasing diagnostic accuracy.Case presentationWe report on a case of SCN of the eyelid diagnosed with the aid of dermoscopy and RCM. A 14-year-old male patient who presented with a painless yellowish-white papule on his left upper eyelid was previously diagnosed with a common wart. Unfortunately, treatment with recombinant human interferon gel was not effective. To achieve a correct diagnosis, dermoscopy and RCM were performed. The former showed closely grouped multiple yellowish-white clods surrounded by linear vessels, and the latter exhibited hyperrefractile material nests at the dermal–epidermal junction level. The alternative diagnoses were, therefore, excluded because of in vivo characterizations. Subsequent surgical excision, histological examination, and von Kossa staining were performed. Pathology showed hyperkeratosis of the epidermis, a downward-directed basal-layer expansion, and small amorphous basophilic deposits scattered throughout the papillary dermis. The von Kossa staining confirmed calcium deposits in the lesion. An SCN was then diagnosed. During the 6-month follow-up, no relapse was observed.ConclusionsPatients with SCN could benefit from dermoscopy and RCM, which help achieve an accurate diagnosis. Clinicians should consider the possibility of an SCN for an adolescent patient with painless yellowish-white papules.

Evaluation of the one-step nucleic acid amplification assay for detecting lymph node metastasis in patients with cervical and endometrial cancer: A multicenter prospective study

ObjectiveThis multicenter study aimed to evaluate the accuracy of the one-step nucleic acid amplification (OSNA) assay in diagnosing lymph node metastasis (LNM) in patients with cervical and endometrial cancers. MethodsSurgically removed LNs from patients with cervical and endometrial cancer were sectioned at 2-mm intervals along the short axis direction and alternately examined using the OSNA assay and conventional histopathological examination. Ultrastaging (200-μm LN sections) was performed for metastatic LNs using hematoxylin and eosin staining and immunostaining with an anti-CK19 antibody in cases where the OSNA assay and histopathological examination (performed using 2-mm LN sections) results showed discordance. ResultsA total of 437 LNs from 133 patients were included; 61 patients (14%) showed metastasis by histopathological examination, with a concordance rate of 0.979 (95% confidence interval [CI]: 0.961–0.991) with the OSNA assay. The sensitivity and specificity of the OSNA assay were 0.918 (95% CI: 0.819–0.973) and 0.989 (95% CI: 0.973–0.997), respectively. Discordance between the two methods was observed in nine LNs (2.1%), and allocation bias of metastatic foci was identified as the major cause of discordance. ConclusionsThe OSNA assay showed equally accurate detection of LN metastasis as the histopathological examination. We suggest that the OSNA assay may be a useful tool for the rapid intraoperative diagnosis of LN metastasis in patients with cervical and endometrial cancers.

Differential Expression and Utilisation of CD34, EMA, PR, and Inhibin in Differentiating Meningioma, Haemangioblastoma, and Solitary Fibrous Tumour: A Cross-sectional Study

Introduction: Mesenchymal tumours of the Central Nervous System (CNS) include meningioma, haemangioblastoma, and Solitary Fibrous Tumour (SFT). Although conventional histopathological examination remains the mainstay in diagnosis, Immunohistochemistry (IHC) plays an important role in CNS tumours, owing to its diagnostic significance and prognostic implications. Aim: To evaluate whether combined use of inhibin, Epithelial Membrane Antigen (EMA,) Progesterone Receptor (PR) and CD34 improves diagnostic accuracy in meningeal and non meningeal mesenchymal tumours of CNS. Materials and Methods: This was a cross-sectional retrospective observational study conducted in the Department of Pathology, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India, October 2022 to December 2022 in which all brain tumour specimens diagnosed as meningeal or non meningeal mesenchymal tumours from January 2022 to August 2022 were included in the present study. Sections were immunostained with an IHC panel of EMA, PR, inhibin, CD34, and Ki67. The immunohistochemical expression of EMA, PR, inhibin, and CD34 was expressed as frequencies and percentages, along with sensitivity and diagnostic accuracy evaluation. Ki67 was assessed in the hotspots and expressed as a percentage. The data were presented as frequencies and percentages. Results: A total of 60 patients were included, with an age range from 12 to 73 years and a mean age of 48.2 years. Female predominance, constituted 40 (67.5%) of the study population. Out of the total cases, 49 (81.7%), 9 (15%), and 2 (3.3%) cases were of meningioma, haemangioblastoma, and SFTs, respectively. Among the 49 cases of meningioma, the majority 47 (96%) showed diffuse and strong staining for EMA with a cytoplasmic staining pattern±membranous accentuation. PR was expressed in 45 (92%) of the meningiomas. Inhibin was diffusely expressed in stromal cells of haemangioblastoma 9 (100%). A characteristic cytoplasmic positivity of CD34 was observed in both cases 2 (100%) of SFT. The Ki67 index of anaplastic and atypical meningiomas was 15%, while all other tumours had a very low proliferation index of 2%. Conclusion: The IHC panel comprising CD34, EMA, and inhibin was useful in the differential diagnosis of both meningeal and non meningeal mesenchymal tumours in the CNS, even in cases with overlapping histomorphological features. Inhibin had a very high sensitivity and diagnostic accuracy for the diagnosis of haemangioblastomas. EMA and PR also had fairly good sensitivity and diagnostic accuracy for the diagnosis of meningiomas. CD34 had good sensitivity but very poor diagnostic accuracy for the diagnosis of SFT.

Culture filtrate supplementation can be used to improve Mycobacterium tuberculosis culture positivity for spinal tuberculosis diagnosis.

Culture remains the gold standard to diagnose spinal tuberculosis (STB) despite the paucibacillary nature of the disease. Current methods can take up to 42 days to yield a result, delaying the ability to rapidly detect drug resistance. Studies have demonstrated the use of supplementation with culture filtrate (CF) from an axenic culture of Mycobacterium tuberculosis (Mtb) as a source of growth factors to improve culture rates. Our objective was to test a modified culture assay, utilizing CF supplemented media (CFSM), to improve culture positivity rates for suspected STB. Twelve patients with suspected STB were assessed by conventional culture (BACTEC™ MGIT 960), GeneXpert™ and standard histopathological examination. Spinal biopsies were taken from areas of diseased vertebral tissue or abscess, predetermined from MRI. Additional biopsies were obtained to assess CFSM for improved detection and faster culture of Mtb. All cases were diagnosed as STB and treated empirically for tuberculosis based on either bacteriological evidence (GeneXpert™, MGIT and/or CFSM positive), or based on clinical presentation. 5 specimens (45.45%) were positive for Mtb DNA as detected by GeneXpert™ and 1 specimen (8.33%) was cultured using MGIT (time to detection; 18 days). CFSM was able to culture 7 specimens (58.3%), with all CFSM positive specimens yielding a culture within 14 days. Two samples were positive only using the CFSM assay pointing to additional yield for diagnostic workup. Modification of standard culture can improve detection of Mtb and reduce time to positivity in individuals with STB where culture material is a requirement.

Cytokine/Chemokine assessment as a complementary diagnostic tool for inflammatory skin diseases.

Inflammatory skin conditions are the 4th leading cause of non-fatal health burden in the general population worldwide. The diagnosis of skin lesions due to systemic drug reactions, viral or bacterial exanthems, or in patients with psoriasis, atopic dermatitis or contact dermatitis is often difficult and relies heavily upon conventional histopathologic examination. Conversely, it is widely accepted that the cutaneous profile of inflammatory markers, or 'inflammatory signature', is differentially expressed in various skin conditions. In this pilot study, we investigated the possibility of inflammatory skin disease diagnosis from an immunological perspective in small punch biopsies. We collected lesional and perilesional punch biopsies from 139 patients suffering from a variety of inflammatory skin conditions and attending the Dermatology Department at the Princess Alexandra Hospital in Brisbane, Australia. Using bead-based immunoassays we were able to measure 13 out of 17 inflammatory markers from a pre-selected multi-analyte panel and to detect significant differences between lesional and perilesional biopsies from each individual patient. Hierarchical and unbiased clustering methods based on inflammatory signatures grouped psoriasis and atopic dermatitis lesions into individual clusters in contrast to other skin conditions, highlighting the potential of inflammatory signatures to be used as diagnostic differentiators and to inform alternative targets in anti-inflammatory treatment strategies.

A study on the placenta in stillbirth: an evaluation of molecular alterations through next generation sequencing

IntroductionPlacental dysfunction is one of the most common causes of Intrauterine Fetal Demise (IUFD). Due to its characteristics, the placenta may be the target of molecular research aimed to investigate potential causes of IUFD. In the literature, there are no studies on human placentas that have investigated possible associations between somatic mutations and the occurrence of IUFD. The aim of this study was to identify the presence of gene mutations in placental tissues in a series of cases of IUFD and to evaluate potential correlations with placental microscopic findings. Materials and methodsThirty-seven samples of formalin-fixed and paraffin-embedded placental tissues were retrospectively selected from pregnancies ending in IUFD between 23rd to 40th week. Six control placentas of physiological pregnancies were included as controls. After sampling, made according to standardized protocol and conventional histopathological examination, placental tissues were subjected to DNA extraction and sequencing by means of Next Generation Sequencing with a 56-gene panel. ResultsThe most frequent mutation observed in 32/37 IUFD cases (86.5%) and absent in any of the 6 control placentas was in c-KIT gene, which is implicated in placental tissue differentiation. However, no significant correlation was found between the presence of individual gene mutations and placental histopatological findings. DiscussionAs the present study found an elevated frequency of c-KIT mutation in IUFD, it further supports the hypothesis that c-KIT is involved in abnormal tissue differentiation leading to altered placental vascularization and function.

Ultrafast 3D histological imaging based on a minutes-time scale tissue clearing and multidirectional selective plane illumination microscopy.

Conventional histopathological examinations are time-consuming and labor-intensive, and are insufficient to depict 3D pathological features intuitively. Here we report an ultrafast 3D histological imaging scheme based on optimized selective plane illumination microscopy (mSPIM), a minutes-time scale clearing method (FOCM), and a deep learning-based image enhancement algorithm (SRACNet) to realize histological preparation and imaging of clinical tissues. Our scheme enables 1-minute clearing and fast imaging (up to 900 mm2/min) of 200 µm-thick mouse kidney slices at micron-level resolution. With hematoxylin and eosin analog, we demonstrated the detailed 3D morphological connections between glomeruli and the surrounding tubules, which is difficult to identify in conventional 2D histology. Further, by the preliminary verification on human kidney tissues, this study will provide new, to the best of our knowledge, feasible histological solutions and inspirations in future 3D digital pathology.

Clear Cell Neoplasms of Salivary Glands: A Diagnostic Challenge.

This review focuses on the heterogenous group of clear cell neoplasms of salivary glands and attempts to identify major differential diagnostic features. Within the head and neck region, clear cells are found most commonly in salivary gland tumors, but may also be seen in tumors of squamous or odontogenic epithelial origin, primary or metastatic carcinomas, benign or malignant melanocytic lesions, or benign or malignant mesenchymal tumors. Clear cells occur fairly commonly among a wide variety of salivary gland neoplasms, but mostly they constitute only a minor component of the tumor cell population. Clear cells represent a major diagnostic feature in two salivary gland neoplasms, epithelial-myoepithelial carcinoma and hyalinizing clear cell carcinoma. In addition, salivary gland neoplasms composed predominantly of clear cells could also include clear cell variants of other salivary neoplasms, such as mucoepidermoid carcinoma and myoepithelial carcinoma, but their tumor type-specific histologic features may only be available in limited nonclear cell areas of the tumor. Diagnosing predominantly clear cell salivary gland tumors is difficult because the immunoprofiles and morphologic features may overlap and the same tumor entity may also have a wide range of other histologic presentations. Many salivary gland tumors are characterized by tumor type-specific genomic alterations, particularly gene fusions of the ETV6 gene in secretory carcinoma, the MYB and MYBL1 genes in adenoid cystic carcinoma, the MAML2 gene in mucoepidermoid carcinoma, the EWSR1 gene in hyalinizing clear cell carcinoma, and others. Thus, along with conventional histopathologic examination and immunoprofiling, molecular and genetic tests may be important in the diagnosis of salivary gland clear cell tumors by demonstrating genetic alterations specific to them.

3D virtual histopathology of cardiac tissue from Covid-19 patients based on phase-contrast X-ray tomography.

For the first time, we have used phase-contrast X-ray tomography to characterize the three-dimensional (3d) structure of cardiac tissue from patients who succumbed to Covid-19. By extending conventional histopathological examination by a third dimension, the delicate pathological changes of the vascular system of severe Covid-19 progressions can be analyzed, fully quantified and compared to other types of viral myocarditis and controls. To this end, cardiac samples with a cross-section of 3.5mm were scanned at a laboratory setup as well as at a parallel beam setup at a synchrotron radiation facility the synchrotron in a parallel beam configuration. The vascular network was segmented by a deep learning architecture suitable for 3d datasets (V-net), trained by sparse manual annotations. Pathological alterations of vessels, concerning the variation of diameters and the amount of small holes, were observed, indicative of elevated occurrence of intussusceptive angiogenesis, also confirmed by high-resolution cone beam X-ray tomography and scanning electron microscopy. Furthermore, we implemented a fully automated analysis of the tissue structure in the form of shape measures based on the structure tensor. The corresponding distributions show that the histopathology of Covid-19 differs from both influenza and typical coxsackie virus myocarditis.

AN OBSERVATIONALANALYTICAL STUDY ON THE PREVALENCE OF FEMALE GENITAL TRACT TUBERCULOSIS AND ITS RELATION TO FEMALE INFERTILITY

INTRODUCTION:Tuberculosis is an infectious disease caused by Mycobacterium species, which are the acid fast bacillus. OBJECTIVES: The present study investigates the prevalence of genital tract tuberculosis among infertile women and its relation to infertility. MATERIALS AND METHOD: The present study is an hospital based analytical and observational study. Department of gynecology and obstetrics, IPGMER hospitals. March 2018 to august 2020. 100 female patients with infertility. CONCLUSION: In our study we reported 11 cases of female with genital tract tuberculosis, diagnosed during laparoscopy, conrmed by CBNAAT, conventional culture method and histopathological examination.

The Value of Sentinel Lymph Node Mapping for the Staging of Node-Negative Colon Cancer: Propensity Score and Mediation Analyses.

Mediation analysis to assess the protective impact of sentinel lymph node (SLN) mapping on prognosis and survival of patients with colon cancer through a more precise evaluation of the lymph node (LN) status. Up to 20% of patients with node-negative colon cancer develop disease recurrence. Conventional histopathological LN examination may be limited in describing the real metastatic burden of LN. Data of 312 patients with stage I & II colon cancer was collected prospectively. Patients were either staged using intraoperative SLN mapping with multilevel sectioning and immunohistochemical staining of the SLN or conventional techniques. The value of the SLN mapping for the detection of truly node-negative patients was assessed using Cox regression and mediation analysis. SLN mapping was performed in 143 patients. Disease recurrence was observed in 13 (9.1%) patients staged with SLN mapping and in 27 (16%) staged conventionally. Five-year overall survival (OS) rate was 82.7% (95% confidence interval [CI], 76.5-89.4%) with SLN mapping compared with 65.8% (95% CI, 58.8-73.7%). Five-year cancer-specific survival (CSS) was 95.1% (95% CI, 91.3-99.0%) with SLN mapping compared with 92.5% (95% CI, 88.0-97.2%). Node-negative staging with SLN mapping was associated with significantly better OS (hazard ratio [HR], 0.64; 95% CI, 0.56-0.72; P < 0.001) and CSS (HR, 0.49; 95% CI, 0.39-0.61; P < 0.001) in multivariate analysis. Mediation analysis confirmed a direct protective effect of SLN mapping on OS (HR, 0.78; 95% CI, 0.52-0.96; P < 0.01) and disease-free survival (DFS) (HR, 0.75; 95% CI, 0.48-0.89; P < 0.01). Staging performed by SLN mapping with multilevel sectioning provides more accurate results than conventional staging. The observed clinically relevant and statistically significant benefit in OS and DFS is explained by a more accurate detection of positive LN by SLN mapping.

Lipidomic analysis and diagnosis of glioblastoma multiforme with rapid evaporative ionization mass spectrometry.

Glioblastoma multiforme (GBM) is the most common and aggressive brain tumor in the central nervous system. GBM patients have a very low 5-year survival rate and most of them died within 1 year. Conventional histopathological examination for GBM diagnosis is complicated and time-consuming, which always blocks the development of more precise and effective treatments in resection operation. Rapid evaporative ionization mass spectrometry (REIMS) is a MS technique in clinical medicine research, which combines the common diathermy device with MS to acquire the lipid profiles of tissue specimens for lipidomic analysis and real-time tumor diagnosis. In this study, the REIMS method employing bipolar forceps was optimized and validated for high-throughput lipidomics and diagnosis of GBM for the first time. Total 42 lipid metabolites were tentatively identified and 12 out of 13 lipid biomarkers showed higher intensities in GBM, which were consistent with previous studies. After this, a statistic model was built with the lipidomic data for the diagnosis of GBM tumor in real-time. The diagnostic accuracy (94.74%), sensitivity (95.38%), and specificity (93.33%) were evaluated with histopathology validated brain tissue specimens that were not used in the training set. The proposed REIMS method for the lipidomic-analysis and diagnosis of GBM tumor provides a new direction for MS-based lipidomics and precision medicine and might be used to guide surgeons to precisely resect the GBM tissue and keep the normal brain tissue in operation.

Nonlinear optical microscopy is a novel tool for the analysis of cutaneous alterations in pseudoxanthoma elasticum

Pseudoxanthoma elasticum (PXE, OMIM 264800) is a rare autosomal recessive disorder with ectopic mineralization and fragmentation of elastin fibers. It is caused by mutations of the ABCC6 gene that leads to decreased serum levels of inorganic pyrophosphate (PPi) anti-mineralization factor. The occurrence of severe complications among PXE patients highlights the importance of early diagnosis so that prompt multidisciplinary care can be provided to patients. We aimed to examine dermal connective tissue with nonlinear optical (NLO) techniques, as collagen emits second-harmonic generation (SHG) signal, while elastin can be excited by two-photon excitation fluorescence (TPF). We performed molecular genetic analysis, ophthalmological and cardiovascular assessment, plasma PPi measurement, conventional histopathological examination, and ex vivo SHG and TPF imaging in five patients with PXE and five age- and gender-matched healthy controls. Pathological mutations including one new variant were found in the ABCC6 gene in all PXE patients and their plasma PPi level was significantly lower compared with controls. Degradation and mineralization of elastin fibers and extensive calcium deposition in the mid-dermis was visualized and quantified together with the alterations of the collagen structure in PXE. Our data suggests that NLO provides high-resolution imaging of the specific histopathological features of PXE-affected skin. In vivo NLO may be a promising tool in the assessment of PXE, promoting early diagnosis and follow-up.

5-ALA-assistant automated detection of lymph node metastasis in gastric cancer patients

Background5-aminolevulinic acid (5-ALA) has been utilized for cancer diagnosis as a fluorescence probe. We have reported the feasibility of 5-ALA-induced protoporphyrin IX (PpIX) fluorescence for detecting lymph node (LN) metastasis in gastrointestinal malignancies. However, a major barrier to the fluorescence diagnosis has been that the evaluation has been highly dependent on the observers. In this study, we examined the validity of a developed device for automated detection without subjectivity.MethodsGastric cancer patients who received oral administration of 5-ALA (20 mg/kg) prior to surgery were enrolled. For a total of 323 LNs obtained from 64 patients, the diagnostic results of the device were compared to those of conventional histopathological examination based on hematoxylin-and-eosin-stained slides. The accuracy with the device was compared to that of stereoscopic detection with conventional fluorescence microscopy for 211 LNs from 42 patients. We used two types of image processing that we previously developed to eliminate autofluorescence of background tissues: differential and ratio methods.ResultsFor detection of metastasis in 323 LNs, the areas under the receiver operating characteristic curves with the differential method and ratio method were 0.921 and 0.909, respectively. The sensitivity, specificity, and accuracy with the differential method were 78.0%, 96.8%, and 94.4%; while those with the ratio method were 78.0%, 96.1%, and 93.8%, respectively. In 211 LN analysis, the diagnostic accuracy with the device was comparable to that of stereoscopic examination.ConclusionOur device for automated detection of LN metastasis using 5-ALA can be a useful tool for intraoperative diagnosis.

Multiclass classification of autofluorescence images of oral cavity lesions based on quantitative analysis.

BackgroundOral cancer is one of the most common diseases globally. Conventional oral examination and histopathological examination are the two main clinical methods for diagnosing oral cancer early. VELscope is an oral cancer-screening device that exploited autofluorescence. It yields inconsistent results when used to differentiate between normal, premalignant and malignant lesions. We develop a new method to increase the accuracy of differentiation.Materials and methodsFive samples (images) of each of 21 normal mucosae, as well as 31 premalignant and 16 malignant lesions of the tongue and buccal mucosa were collected under both white light and autofluorescence (VELscope, 400-460 nm wavelength). The images were developed using an iPod (Apple, Atlanta Georgia, USA).ResultsThe normalized intensity and standard deviation of intensity were calculated to classify image pixels from the region of interest (ROI). Linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA) classifiers were used. The performance of both of the classifiers was evaluated with respect to accuracy, precision, and recall. These parameters were used for multiclass classification. The accuracy rate of LDA with un-normalized data was increased by 2% and 14% and that of QDA was increased by 16% and 25% for the tongue and buccal mucosa, respectively.ConclusionThe QDA algorithm outperforms the LDA classifier in the analysis of autofluorescence images with respect to all of the standard evaluation parameters.