Histology Laboratory Research Articles

National society for histotechnology workload study

ABSTRACT In June and July 2021, the National Society for Histotechnology (NSH) conducted an online survey designed to assess productivity and staffing in the clinical histology laboratory. The Productivity Benchmarking Survey was developed by the NSH Quality Management Committee. The survey of histology professionals provides critical data that may be used to inform staffing decisions and develop a quality management program. To create usable data the findings were segmented by a range of variables including facility type and size, and productivity related to grossing, embedding, microtomy, and ancillary duties. This study draws a connection between perceived operational efficiencies and the presence of a program or process used to assess employees quality performance for grossing,embedding, and microtomy. A followup survey is planned to further understand these particular employee assessment programs or processes.

Histomorphological Features of Ovarian Neoplasms and Expression of p53 and WT1 in Surface Epithelial Tumours: A Cross-sectional Study

Introduction: Ovary is the most common site of neoplastic and non neoplastic lesion, can present in childhood to postmenopausal age group and remain the most lethal of all gynaecological malignancies. Tumour Protein (p53) gene mutations or deletions are most common in ovarian carcinoma. However, Wilms' Tumor gene1 (WT1) and p16 expression are also seen in serous ovarian carcinoma. These Immunohistochemistry (IHC) marker are useful in the differential diagnosis of serous ovarian carcinomas. Aim: To study histomorphology of ovarian neoplasm along with expression of p53 and WT1 in surface epithelial tumours and to assess the prevalence of various ovarian neoplasms in different age groups. Materials and Methods: This cross-sectional study was conducted in Department of Pathology, Sarojini Naidu Medical College, Agra, Uttar Pradesh, India, over a period of two years (from September 2018-September 2020). A total of 78 ovarian specimen received in histology laboratory was studied for gross and microscopic features. The p53 and WT1 IHC expression pattern was studied in surface epithelial tumours. Statistical significance was calculated in relation to p53 and WT1 expression with histological type and grade of tumour using Chi-square test. Results: A total of 78 cases were studied, out of which benign tumours were the most common 40 (51.3%) cases, followed by malignant tumours 32 (41.0%) cases and borderline malignancy 6 (7.7%) cases. The most common benign lesions were mucinous cystadenoma 20 (50%) cases. Serous carcinomas were most common malignant tumours 19 (59.3%) cases followed by Germ Cell Tumours (GCT) 5 (15.6%) cases. All benign and borderline epithelial ovarian tumours were found p53 and WT1 negative. Out of 22 cases of malignant surface epithelial tumour, 14 (63.6%) and 12 (54.4%) were positive for p53 and WT1 respectively and all were serous carcinomas. Conclusion: Ovarian lesions present with wide spectrum of histomorphological features. The p53 and WT1 show different rates of expression and staining pattern in various epithelial ovarian carcinomas. Hence routine gross, proper histological examination and correct IHC interpretation is required for specific diagnosis.

Correlation of histological diagnosis and laboratory findings in distinguishing acute appendicitis and lymphoid hyperplasia.

Acute appendicitis (AA) is one of the most frequent causes of abdominal surgery encountered in emergency rooms. However, reactive lymphoid hyperplasia (RLH) is one of the appendicular pathologies often misdiagnosed. It is quite challenging to distinguish between AA and RLH in terms of planning treatment in an emergency. Therefore, this retrospective study aimed to compare the histological and laboratory findings of AA and RLH. The retrospective data included in the study were obtained from patients diagnosed with AA. Complete blood count (CBC) and C-reactive protein (CRP) levels of patients with AA and RLH were compared before the surgery based on the histological diagnosis of the patients. A total of 187 patients who previously underwent appendectomy were included in the study. Histopathological examination revealed that 152 patients (81.3%) were diagnosed with AA, and 35 (18.7%) with RLH. While white blood cell count (P < 0.001), neutrophil (P < 0.001), and neutrophil/lymphocyte ratio (P < 0.001) were found to be significantly higher in those with AA; lymphocyte (P = 0.003) and eosinophil counts (P = 0.033) were detected to be significantly higher in those with RLH. CRP level was also significantly higher in those with AA (P = 0.002). We consider that CBC and CRP levels may be predictive in distinguishing between AA and RLH. We consider that these parameters may be valuable in making a distinction between patients before surgery.

Implementation of a required 3rd-year medical student surgical pathology clinical experience

Few medical schools have required experience ​in surgical pathology during the clinical years. After introducing a pilot and preliminary surgical pathology clinical experience into the curriculum, we initiated a required 3rd-year medical student surgical pathology clinical experience that consisted of a one hour introductory lecture; one hour gross room, histology, and immunohistochemistry laboratory introduction; and one hour of one-on-one case sign-out preceptorship with a subspecialty surgical pathologist within the surgery and obstetrics/gynecology block. Concepts that were covered included specimen processing, intraoperative frozen section consultation, completing specimen requisitions, interpreting synoptic reports, and pTNM staging. Students evaluated the surgical pathologist from 1 to 5 (1 “poor/unhelpful,” 2 “marginal,” 3 “neutral,” 4 “good,” 5 “excellent/useful”). Ten multiple-choice questions (included as part of a perioperative services exam) and attendance were incorporated into students’ perioperative services rotation grade. From 2014 to 2018, 757 students participated in the required 3rd-year surgical pathology clinical experience. Thirty academic subspecialty pathologists acted as preceptors with an average of nine sessions per preceptor per year. Evaluation data from 316 students from 2015 to 2018 showed a mean preceptor rating of 4.8/5 (range 4.0–5.0). Students scored an average of 81% on the surgical pathology portion of the exam (range 21–99% for each question). We successfully implemented a required medical student surgical pathology clinical experience. At the clerkship’s conclusion, students demonstrated understanding of key concepts and rated their preceptorship experience highly.

Evaluation of Histomorphological Spectrum of Skin Lesions at a Teaching Institute in Agra: A Cross-sectional Study

Introduction: The skin is the largest organ system in humans. Different skin diseases consist of non specific, non infectious and infectious diseases to various neoplastic lesions. Dermatological lesions are commonly encountered in all countries and it encompasses a wide spectrum, varies from country to country and various regions within a country and influenced by sex, age and associated systemic disorders, economy, literacy, racial and social customs. The clinical presentation is restricted to only a few changes such as hyperpigmentation, hypopigmentation, macules, papules, nodules and few others. However, the spectrum of histopathology of skin disorders is varied. Accurate diagnosis of skin disorders is of utmost importance as treatment is varied for different skin disorders presenting with the similar clinical lesions. Aim: To study histomorphological spectrum and distribution of skin lesions. Materials and Methods: This hospital-based crosssectional study was conducted in Department of Pathology in collaboration with Department of Skin and Venereal Diseases, Sarojini Naidu Medical College, Agra, Uttar Pradesh, India, over a period of two years from September 2019 to August 2021. The biopsy sample taken from clinically diagnosed skin lesions sent to histology laboratory in 10% formalin. Sample was fixed in 10% neutral buffered formalin over a period of 12 to 24 hours. Paraffin wax blocks were made and 3-4 micrometer sections were taken and stained with Haematoxylin and Eosin (H&amp;E) stain and histology was studied under microscope. Special stains such as Ziehl-Neelsen (Z-N) stain and Fite stain were done, whenever required. Data was entered in Microsoft Excel excel and descriptive data was obtained. Results: Out of total 105 cases, the maximum patients were of 21-30 years of age group 30 (28.57%) cases followed by 11-20 years of age group 25 (23.81%) cases with male and female ratio M:F=1.5:1. In this study, out of total 105 cases, the most common site involved was trunk in 42 (42%) cases followed by upper limb in 25 (23.81%) cases. The most common skin lesions were non infectious 70 (66.67%) cases, followed by infectious disease 22 (20.95) cases. In non infectious papulosquamous diseases group found 25 (23.81%) cases followed by vesiculobullous and vesiculopustular diseases group 24 (22.86%) cases. Infectious diseases were seen in 22 (20.95%) cases, out of which bacterial diseases being most common in 19 (18.10%) cases followed by parasitic infection 2 (1.90%) cases. Conclusion: According to the findings of the present study the major cases of skin lesions requiring biopsy in developing countries is still psoriasis in non infectious papulosquamous diseases group. Tuberculosis and leprosy in infectious diseases category were being the leading cause of morbidity.

Clinical outcomes from the Assessing Donor-derived cell-free DNA Monitoring Insights of kidney Allografts with Longitudinal surveillance (ADMIRAL) study

The use of routine monitoring of donor-derived cell-free DNA (dd-cfDNA) after kidney transplant may allow clinicians to identify subclinical allograft injury and intervene prior to development of clinically evident graft injury. To evaluate this, data from 1092 kidney transplant recipients monitored for dd-cfDNA over a three-year period was analyzed to assess the association of dd-cfDNA with histologic evidence of allograft rejection. Elevation of dd-cfDNA (0.5% or more) was significantly correlated with clinical and subclinical allograft rejection. dd-cfDNA values of 0.5% or more were associated with a nearly three-fold increase in risk development of de novo donor-specific antibodies (hazard ratio 2.71) and were determined to be elevated a median of 91 days (interquartile range of 30-125 days) ahead of donor specific antibody identification. Persistently elevated dd-cfDNA (more than one result above the 0.5% threshold) predicted over a 25% decline in the estimated glomerular filtration rate over three years (hazard ratio 1.97). Therefore, routine monitoring of dd-cfDNA allowed early identification of clinically important graft injury. Biomarker monitoring complemented histology and traditional laboratory surveillance strategies as a prognostic marker and risk-stratification tool post-transplant. Thus, persistently low dd-cfDNA levels may accurately identify allograft quiescence or absence of injury, paving the way for personalization of immunosuppression trials.

Detection of Technical Errors and Construction Optimization for Custom Tissue Arrays at Transplanting and Casting

Purpose: The purpose of this research is to identify causes for the most common typical defects in the production of tissue arrays at the most crucial stages, and to optimize protocols for custom-made tissue arrays at variable input parameters and different final requirements. In this way, we can avoid difficult repairs of tissue arrays, at which, because of re-allocation, major losses can occur, including destruction of the sample due to mechanical and thermal effects.&#x0D; Methods: Seven hundred and twenty tissue array blocks were included, some for clients and others for experimentation and protocol refinement. Human and animal tissue were used. We tested and evaluated configurations, structural stability, consistency, stability, and practical usability of such designed tissue arrays under different laboratory physical conditions.&#x0D; Results: We recognized and analyzed 12 mistakes in the production of tissue arrays in the histology laboratory during the stages of transplantation and casting. At each examined error we found the causes, gave suggestions for prevention, and provided instructions on how to eliminate or prevent those causes.&#x0D; Conclusion: Problems have solutions that are technically feasible but they require individual adjustments of laboratory procedures, time, and learning. This is more difficult or even impossible for automated laboratories, as in the described handmade tissue arrays. However, all laboratories need to produce with maximum quality and precision, which is possible only with addressing problems regularly. The results of this study will contribute to the optimization of custom made tissue array processes.

BECOMING OF THE DEPARTMENT OF HUMAN ANATOMY AT POLTAVA STATE MEDICAL UNIVERSITY SINCE ITS FOUNDATION AND ITS MAJOR ACHIEVEMENTS

In its becoming, Poltava State Medical University has passed a thorny path from the odontology faculty, created on the basis of the Kharkov Medical Academy in 1921, to the leading specialized institution for training medical and healthcare professionals. In 2021 the university has celebrated its 100th anniversary. In 1950, the Department of Human Anatomy moved to a new location and dedicated to the training of medical professionals, research activities. The scope of scientific research conducted at the department covered issues on the structure of glands of the mucous membranes, lymphoid tissue of the initial section of the digestive system, structural peculiarities of autonomic nervous system. The department put a lot of effort to equip microscopic, histological, histochemical and photo laboratories, to create the museum of the department. Scientists developed and manufactured various devices and techniques for conducting morphological studies, which made it possible to publish numerous methodological recommendations, textbooks, and to conduct research for the candidate and doctor degrees. The anatomical museum, refurbished and modernized in 2005 and known even abroad is the subject of honour not only for the department, but for the University as well. The museum is also a matter of utmost interest to young visitors during the Open Days, students from other universities. The Department of Human Anatomy seeks to capitalize on a number of pedagogical experiences and research achievements to transfer knowledge and know-how to young generation of students and scientists.

The "Magnificent Seven Errors" in Forensic Autopsy Practice: The Italian Context.

According to the Italian legal system, forensic autopsies are performed by a medical doctor specialized in legal medicine, otherwise known as a medicolegal expert (MLE), who has a range of very complex responsibilities. Indeed, the quality of forensic autopsy activity is always questioned in courts of law; incorrect assessments are dangerous because they can jeopardize the validity of a criminal investigation and thus affect the outcome so that a real culprit may be acquitted or an innocent person convicted. Nonconformities also discredit the professionalism of the specialist who performs the autopsy. The work of a MLE implies a series of assignments and duties that should be given constant consideration, but when certain aspects of this activity are underestimated or overlooked, this can lead the expert to make mistakes with irreparable consequences for the judicial investigation. In this article, for the first time, we present a summary of seven known errors related to autopsy activity following death by unnatural causes, with the purpose of alerting MLEs who work under the Italian judicial system to the potential dangers of such errors. These relate to: oversights in autopsy technique, incorrect collection of photographic and video material, unauthorized attendance at the autopsy, missing/mistaken reporting at any stage of the forensic activity, failure to notify the party forensic consultant, using histological or toxicological nonaccredited laboratories for forensic activities, and lack of observance of the chain of custody.

Relationship between skin structure and wool traits for Ultrafine and Dohne Merino sheep of Uruguay

The demand in the world market for high quality fine and superfine wool is increasing, responding to greater wool textile industry demands and price incentives. There are few research studies focused on hair follicle population in fine and superfine Merino sheep in Uruguay. This study evaluates the association between wool follicle population and the most relevant economical wool traits. This work was carried out on Ultrafine Australian Merino (UAM) and Dohne Merino (DM) animals (lambs and rams). Phenotypic correlations between the secondary/primary follicles (S/P) and other wool traits (fiber diameter, staple length, clean fleece weight, scoured yield, coefficient of variation of fiber diameter and comfort factor parameter, were performed using histological skin cuts. Two skin samples were extracted from each lamb at two and nine months of age. These were processed at the skin histology laboratory to obtain a histological preparation for further biopsy, being able to differentiate the structures obtained to be evaluated. The results showed in the UAM group, S/P ratio in nine-month-old lambs was greater than two-month-old lambs (31.08 vs 23.12 P&lt; 0.05). For DM group, there were no differences in the S/P ratio between age groups. Phenotypic correlations for wool characteristics obtained were of low to medium or null magnitude in both genotypes.

MA12.02 Factors Associated with Severe COVID-19 Infections in Lung Cancer Patients

Lung cancer patients are at increased risk of developing complications from COVID-19 disease. Real-world data may better identify lung cancer patients who are at an increased risk of developing complications while maximizing the benefit of anti-cancer therapy. We conducted a retrospective cohort study using the VA COVID-19 Shared Data Resource (CSDR), VA Cancer Registry, and VA Corporate Data Warehouse (CDW), which centralizes electronic health data for patients seen at VA facilities nationwide. We included patients with a diagnosis of lung cancer between October 1, 2015 and December 1, 2020, and a diagnosis of COVID-19 between February 2, 2020 and December 1, 2020. Serious SARS-CoV-2 infection was defined as the occurrence of any of the following within 2 weeks after diagnosis: (a) hospitalization, (b) ICU admission, (c) utilization of respiratory support (mechanical ventilation or intubation). Data abstracted included: age at COVID-19 diagnosis, gender, race, ethnicity, urban status, date of diagnosis of lung cancer, histology, stage, cancer treatment, individual comorbidities and most recent laboratory results prior to COVID-19 diagnosis. Patients were stratified into 3 groups: mild/moderate COVID-19 infection, serious, but non-fatal infection, fatal infection. Differences in categorical variables, were assessed using χ2 test, while Kruskal-Wallis rank sum test was used for continuous variables. Multivariable logistic regression models were fit relative to the outcomes of interest, i.e., serious SARS-CoV-2 infection and death from SARS-CoV-2 infection. We identified 352 lung cancer patients with COVID-19. Of these, 54 (15.3%) had severe disease and 34 others (9.7%) died. Patients who had fatal or severe infection were older than those with mild/moderate infection (median age: 74.1 and 73.8 yrs vs. 72.1 yrs; p=0.01). Patients who suffered a fatal or severe infection were also more likely than patients with mild/moderate infection to exhibit elevated creatinine levels (50.0% and 31.5%, vs. 20.5%, p = 0.003) and low hemoglobin levels (67.6% and 51.9%, vs. 39.0%, p = 0.003). In addition, 70.6% of patients who died within two weeks of SARS-CoV-2 infection suffered from diabetes, compared to 40.2% of patients with mild/moderate infection.On multivariable logistic regression, variables associated with increased odds of severe infection or death were: age (OR: 1.07; 95% CI 1.03-1.12; p=0.002), stage IV (6.06; 1.19-30.84; 0.03), elevated creatinine (2.29; 1.2-4.39; 0.01), anemia (2.02, 1.07-3.83; 0.03). Type of cancer treatment, recent surgery or radiation, chronic obstructive pulmonary disease, hypertension, diabetes, acute myocardial infection were not associated with an increased risk of severe/fatal infection. Factors associated with fatal infection included checkpoint inhibitor therapy (OR: 7.06; 95% CI, 1.21-41.11; p=0.03), diabetes (3.53; 1.9-11.46; 0.04) and abnormal creatinine level (4.48; 1.52-13.15; 0.006). Almost 25% of lung cancer patients with COVID-19 infection developed complications or died. Increasing age, stage IV disease, abnormal kidney function and low hemoglobin level were associated with a severe/fatal SARS-CoV-2 infection, while checkpoint inhibitor therapy, diabetes and abnormal creatinine levels were associated with increased mortality from COVID-19 disease.

Comparative Analysis of Eosin-based Fluorescence Microscopy of Non-neoplastic Breast Tissue and Fibroadenoma

Background: Fibroadenoma (FA) is one of the most frequently diagnosed benign neoplasm in women. Various researches have reported increased risk of breast cancer in females with FA. It stems from the proliferation of epithelial and stromal contents of the terminal duct lobular units (TDLU`S) of breast tissue, that are the primary sites for the histopathologic assessment which is the gold standard for the diagnosis of disease. However, this method is subjective and possess interobserver variability. Therefore, new quantitative methods are required to aid in diagnosis. Hence we evaluated fluorescence light intensity and its use in histopatholgic evaluation.&#x0D; Aim: The goal of this research was to compare and quantify red and green fluorescence light intensities of ductal cells and stroma of non-neoplastic breast tissue with fibroadenomatous tissue.&#x0D; Method: A cross-sectional study was done in the Cell biology and histology lab of Ziauddin University. 44 slides of normal breast tissue and 44 slides of diagnosed fibroadenomatous tissue were taken from Dr Ziauddin Hospital, North Campus. Hematoxylin and eosin (H&amp;E) staining of the slides were done following standard protocols. On microscopic examination, the changes in light intensities of ductal cells and stroma of normal breast tissue and fibroadenoma were quantified using dual channel fluorescence microscopy using Nikon NIS imaging software.&#x0D; Results: The results demonstrated statistically significant increase (p-value &lt;0.05) in mean red (37.22±5.9) and green (22.47±6.6) light intensity of stroma in FA when compared with red (32.71± 6.7) and green (17.01±4.3) light intensity of normal breast tissue. Whereas, R/G ratio for normal tissue was higher (1.95±0.11) than R/G for FA (1.74±0.37) with a p value of &lt;0.05. Similarly, for ductal cells; statistically significant (p value &lt;0.05) increase in mean red (38.86±5.4) and green (15.54±2.51) light intensity for FA was found when compared with red (29.62±1.89) and green (12.60±1.67) intensity of normal tissue. R/G ratio for FA (2.5±0.24) was compared to be higher than normal tissue (2.36±0.3) with a p value of &lt;0.05.&#x0D; Conclusion: The study suggests that fluorescence microscopy combined with quantitative assessment fluorescence light intensities may may be a helpful tool for histomorphic evaluation of the breast tissue.

Digital Pathology in Cameroon.

PURPOSECancer is becoming increasingly prevalent among the group of treatable diseases in African countries. There is a shortage of clinicians and pathologists available for cancer diagnosis and treatment. These limited resources must be efficiently used to maximize the number of patients treated. One of the critical factors in treatment efficiency is the correct and timely diagnosis of specimens by pathologists. However, there is currently a significant shortage of cancer care clinicians in Africa and an even more considerable shortage of pathologists. This article presents an example in which telepathology was used to mitigate the lack of pathologists in Cameroon.METHODSThe telepathology workaround was implemented in a district hospital based in Cameroon's Adamawa region, where a European surgeon provides cancer treatment. A small histology laboratory there is run by one histotechnologist who processes surgical biopsies into histology slides. As there are no pathologists on site, these slides are digitally scanned using a mobile phone and a whole slide imaging (WSI) scanner. The slides are then shared electronically with a volunteering pathologist in Europe who provides a diagnostic report.RESULTSFrom 2018 to July 2019, specimens for 101 patients were photographed through an iPhone connected to a microscope eyepiece producing several individual images per specimen. From July 2019 to December 2020, slides from 282 patients were scanned using WSI and digitally transmitted.CONCLUSIONWSI on hematoxylin and eosin histology slides for remote diagnosis can increase cancer treatment efficacy and reduce overtreatment of tumors clinically suspicious for malignancy in under-resourced countries with a lack of pathologists.

Synthetic Antigen Controls for Immunohistochemistry.

Immunohistochemistry (IHC) assays provide valuable insights into protein expression patterns, the reliable interpretation of which requires well-characterized positive and negative control samples. Because appropriate tissue or cell line controls are not always available, a simple method to create synthetic IHC controls may be beneficial. Such a method is described here. It is adaptable to various antigen types, including proteins, peptides, or oligonucleotides, in a wide range of concentrations. This protocol explains the steps necessary to create synthetic antigen controls, using as an example a peptide from the human erythroblastic oncogene B2 (ERBB2/HER2) intracellular domain (ICD) recognized by a variety of diagnostically relevant antibodies. Serial dilutions of the HER2 ICD peptide in bovine serum albumin (BSA) solution are mixed with formaldehyde and heated for 10 min at 85 °C to solidify and cross-link the peptide/BSA mixture. The resulting gel can be processed, sectioned, and stained like a tissue, yielding a series of samples of known antigen concentrations spanning a wide range of staining intensities. This simple protocol is consistent with routine histology lab procedures. The method requires only that the user have a sufficient quantity of the desired antigen. Recombinant proteins, protein domains, or linear peptides that encode relevant epitopes may be synthesized locally or commercially. Laboratories generating in-house antibodies can reserve aliquots of the immunizing antigen as the synthetic control target. The opportunity to create well-defined positive controls across a wide range of concentrations allows users to assess intra- and inter-laboratory assay performance, gain insight into the dynamic range and linearity of their assays, and optimize assay conditions for their particular experimental goals.

ENDOMETRIAL BIOPSIES PATTERNS AMONG WOMEN ATTENDING MOI TEACHING AND REFERRAL HOSPITAL, KENYA

Background: Endometrial diseases are among the most common gynecological disorders affecting women both globally and locally;accounting for 60% of global maternal deaths.Kenya is ranked thirteenth out of 181 countries with the highest maternal mortality globally. Endometrial disorders such as hyperplasia's, neoplastic, inflammatory and pregnancy related conditions of the endometrium have been reported to increase with demographic traits such as maternal age and parity. Objective: To determine histopathological patterns of endometrial biopsies among women attending Moi Teaching and Referral Hospital,Kenya. Methods: This was a retrospective laboratory-based study where 121 banked endometrial biopsy blocks collected from participants aged 19 to 70 years between August 2014 and August 2016 were retrieved, re-sectioned and stained using routine histological (Hematoxylin and Eosin) stains in the histology laboratory before being sent to the pathologist for examination. Sociodemographic and reproductive history data were collected from the medical records. Descriptive statistical techniques such as cross-tabulation were used.The median age of all the study participants was 44 years. Results: Most of the study participants (39.2%;n=38) presented with simple endometrial hyperplasia.The endometrial patterns were categorized as Hyperplasias (50.4%; n=61), Inflammatory disorders (7.4%; n=9), Neoplastic Disorders (14.9%;n=18),Pregnancy related disorders (20.7%;n=25) and other patterns (6.6%;n=8) Conclusion: Majority of the study participants presented with simple endometrial hyperplasia followed by complex hyperplasia and adenocarcinoma,respectively.

Microscopic Study of Mechanoreceptors and Chemoreceptors of Anterior and Posterior Ends of Toxocara Canis Using Scanning Electron Microscopy and Light Microscope.

The present study investigated the fine structure of amphids and phasmids, cuticle, muscles, and digestive tracts of Toxocara canis using optical and electron microscopy, hematoxylin-eosin (H&amp;amp;E) staining, and other specific stains. A number of 38 adult T.canis worms were obtained from the animal shelter of Urmia, and their small intestines were fixated in acidified formal alcohol and 10% formalin solutions. The anterior and posterior parts of male and female T.canis worms were prepared and cut at a thickness of 4-5 &amp;mu;m according to the conventional method in the histological laboratory. The samples were then stained using H&amp;amp;E and specific periodic acid-Schiff, Masson&amp;#39;s trichrome, and Orcein staining. The structure of amphid (anterior), phasmid (posterior), cuticle, muscles, and digestive tracts of male and female worms were studied under light microscopy. Basal, intermediate, cortex, and cuticle surface coating of the parasite were visible. Alae were also observed as the thickenings in the cuticle. The muscle layer structure consists of non-branched cylindrical cells. The intestinal tract is composed of cuticular cogs, the esophagus is of filamentous-muscular structure, and the intestine is made of columnar epithelial tissue with microvilli and glycocalyx. The amphid structure consisted of cuticular protrusions with penetrations of the cephalic framework into their inner layers. Phasmid structure also includes protrusions in the cuticle and invagination of sensory neurons. It was concluded that for the most part, the histological structure of the cuticle can be studied by optical microscopy. The muscle structure in this parasite was very similar to the skeletal muscle in mammals. Furthermore, the epithelial structure of the intestine in this parasite was largely similar to the intestinal epithelium in mammals. Finally, regarding the amphid and phasmid structures, it was observed that they were protrusions covered by cuticles where neural, filamentous, and muscular structures were the core of these protrusions.

Platelet Programmed Cell Death Ligand 1 (pPDL-1) Is a Prognostic Marker in Advanced Lung Cancer

Introduction. PDL-1 (Programmed Cell Death Ligand-1) expressed on non small cell lung cancer (NSCLC) cells mediates immune escape, and inhibition of PDL-1/PD-1 interaction with blocking antiboides induces long-term responses in a subpopulation of patients with metastatic NSCLC. PDL-1 immunohistochemistry is a controversial means of predicting responders, and further biomarkers are urgently needed. Herein, we report on the phenomenon of PDL-1 expressed on the surface of platelets (platelet PDL-1, pPDL-1) in patients with advanced NSCLC.Methods. Blood samples were prospectively collected from patients, and platelet-expressed PDL-1 expression was measured by flow cytometry utilizing flow monoclonal antibodies clones MIH1 and 28-8. Correlations of pPDL-1 expression levels with histopathological data, laboratory data, and survival were analyzed.Results. 79 NSCLC patients (53 with adenocarcinoma and 26 with squamous cell carcinoma) and 42 healthy controls were analyzed. 61 patients (77.2%) had stage IV disease. Platelets derived from healthy controls expressed little if any pPDL-1. Mean pPDL-1 expression in tumor patients was significantly increased compared with controls. PPDL-1 was not correlated with histological subtypes, mutational status, smoking history, clinical stage or laboratory data. However, overall survival (OS) and progression free survival (PFS) were significantly reduced in patients with a high (above median) percentage of PDL-1-positive platelets as compared with low pPDL-1(OS: 29 vs. 45 months, p = 0.027, PFS: 6 vs. 9 months, p = 0.036). The accuracy of our system was validated by calculating the area under the receiver-operator characteristic (ROC) curve. Here, platelet-derived PDL-1 expression was also significantly associated with a reduced OS (AUC=0.719, 95 CI: 0.587- 0.851) and PFS (AUC=0.768, 95 CI: 0.59- 0.89).Conclusion. To our knowledge, this is the first report of PDL-1 expression on platelets. In NSCLC patients, high pPDL-1 - similar to tumor PDL-1 - is clearly prognostic. The functional significance and predictive value of pPDL-1 remains to be established. DisclosuresNo relevant conflicts of interest to declare.

Platelet count as a screening tool for compensated cirrhosis in chronic viral hepatitis.

BACKGROUNDSimple tools for clinicians to identify cirrhosis in patients with chronic viral hepatitis are medically necessary for treatment initiation, hepatocellular cancer screening and additional medical management.AIMTo determine whether platelets or other laboratory markers can be used as a simple method to identify the development of cirrhosis.METHODSClinical, biochemical and histologic laboratory data from treatment naive chronic viral hepatitis B (HBV), C (HCV), and D (HDV) patients at the NIH Clinical Center from 1985-2019 were collected and subjects were randomly divided into training and validation cohorts. Laboratory markers were tested for their ability to identify cirrhosis (Ishak ≥ 5) using receiver operating characteristic curves and an optimal cut-off was calculated within the training cohort. The final cut-off was tested within the validation cohort.RESULTSOverall, 1027 subjects (HCV = 701, HBV = 240 and HDV = 86), 66% male, with mean (standard deviation) age of 45 (11) years were evaluated. Within the training cohort (n = 715), platelets performed the best at identifying cirrhosis compared to other laboratory markers [Area Under the Receiver Operating Characteristics curve (AUROC) = 0.86 (0.82-0.90)] and sensitivity 77%, specificity 83%, positive predictive value 44%, and negative predictive value 95%. All other tested markers had AUROCs ≤ 0.77. The optimal platelet cut-off for detecting cirrhosis in the training cohort was 143 × 109/L and it performed equally well in the validation cohort (n = 312) [AUROC = 0.85 (0.76-0.94)].CONCLUSIONThe use of platelet counts should be considered to identify cirrhosis and ensure optimal care and management of patients with chronic viral hepatitis.

Succeeding in a pandemic: Online lectures and in‐person laboratory teaching maintained exam scores in the anatomical sciences

IntroductionThe COVID‐19 pandemic forced many schools to adopt more online teaching than they would normally have chosen for 2020. While many medical schools canceled laboratory teaching sessions in the anatomical sciences, a minority maintained laboratory teaching, including ours. Due to the pandemic, students had far fewer opportunities to interact with their peers and faculty, and students may have experienced greater non‐academic stress, all of which would be expected to negatively impact student academic performance. Also, some faculty members feel that virtual vs. in‐person lectures hinder student learning. Taken together, these issues led to significant concern about lower academic performance by first‐year medical students in 2020. Our curriculum changed very little between 2019 and 2020, presenting the opportunity to assess the impact of online lectures while keeping the laboratory component essentially unchanged. We hypothesized that maintaining laboratory‐based teaching minimized the negative effects of online lectures and student stress in 2020.MethodsAcademic performance in the anatomical sciences between 2019 (98 students) and 2020 (143 students) was compared using scores from five written and five laboratory exams from Medical Gross Anatomy (MGA) and four histology laboratory exams from the Foundations of Medicine I course (FOM‐I). In addition, course evaluation data for the two courses between 2019 and 2020 was compared.ResultsThe exam scores between 2019 and 2020 differed significantly overall (one‐way PERMANOVA for non‐parametric data, p = 0.003, 9999 permutations). In pairwise comparisons of specific exams between the two years, students in 2020 performed significantly better on one gross anatomy written exam, one gross anatomy laboratory exam, and one histology laboratory exam, whereas 2019 students performed significantly better on one histology laboratory exam (Mann‐Whitney U tests, dfs = 239, p‐values = 0.02 or less). The mean scores from student course evaluations for both MGA and FOM‐I were slightly higher in 2020 compared to 2019, although the differences were not statistically significant (Independent Sample t‐tests, dfs = 213, p = 0.23, 0.13, respectively).DiscussionThese results indicate that the change to all‐online lectures with standard in‐person laboratory sessions in 2020 did not harm student academic performance in the anatomical sciences. The one lower exam score in histology in 2020 may be due to two additional topics being included on that exam in 2020. If the likely extra‐academic stressors that some students may have experienced due to the pandemic (whether from social isolation or direct impact of the disease on family, friends, or themselves) are factored in, the similarity in academic performance in 2019 and 2020 suggests that online lectures paired with in‐person laboratory learning may in fact be beneficial for student academic performance, although this would need further investigation. This study will enable comparisons with medical schools that had entirely virtual courses, with either virtual or no laboratory sessions.This study of human subjects was approved by the Texas A&M University Institutional Review Board.

Sensitive label-free imaging of brain samples using FxClear-based tissue clearing technique

SummaryOptical clearing has emerged as a powerful tool for volume imaging. Although volume imaging with immunostaining have been successful in many protocols, yet obtaining homogeneously stained thick samples remains challenging. Here, we propose a method for label-free imaging of brain slices by enhancing the regional heterogeneity of the optical properties using the tissue clearing principle. We used FxClear, a method for delipidation of brain tissue, to retain a larger proportion of lipids at the white matter (WM). Furthermore, the embedding media affected the contrasts for the lipid-rich or extracellular matrix-rich areas, depending on their chemical properties. Thus, we tailored clearing conditions for the enhancement of the refractive indices (RIs) differences between gray and WM, or several pathological features. RI differences can be imaged using conventional light microscopy or optical coherence tomography. We propose that our protocol is simple, reliable, and flexible for label-free imaging, easily implementable to routine histology laboratory.