Standard Histological Examination Research Articles

IDENTIFICATION AND PATHOLOGY OF STRONGYLUS SPECIES IN NATURALLY INFECTED MULES.‎

Naturally infected large intestine of 16 mules were collected during one year period. All three species of Strongylus were identified: S.vulgaris, S.equinus and S.edentatus. Specimens were processed for standard histological examination. The effect of the parasite on the wall of the intestine ‘included the _ surface epithelium, glandular tissue, muscularis mucosa and part of the submucosa. Changes on the intestinal wall were observed at the site of the parasitic attachement, which included oedema, cellular infilteration with congestion and haemorrhages.

Cytology Techniques Can Provide Insight into Human Placental Structure Including Syncytiotrophoblast Nuclear Spatial Organisation.

The aim of this study was to provide the first systematic description of human placental cytology appearances and to investigate syncytiotrophoblast nuclear organisation patterns using cytology techniques. Term placentas from normal pregnancies were sampled using fine-needle aspiration (FNA) and direct scrapes. Standard histological examination was also performed to exclude pathological changes in the placentas being studied. Both Papanicolaou-stained cytospin preparations and air-dried Giemsa slides from FNA provided high-quality material for cytological assessment with good cellularity. Among the key features of the cytology preparations were villous "microbiopsies" that allowed for the three-dimensional appreciation of villous branching patterns. Cytological appearances, including nuclear characteristics of villous cytotrophoblast and syncytiotrophoblast, were also well demonstrated. In microbiopsies and detached villous trophoblast sheets, complex patterns of syncytiotrophoblast nuclear organisation, not previously described cytologically, were observed, including irregular spacing of nuclei, syncytioplasm windows and linear nuclear arrangements. This study showed that placental cytology (a) provides technically excellent material for cytological evaluation, (b) confirms the presence of complex nuclear organisational patterns in the syncytiotrophoblast by eliminating the possibility of tangential sectioning artefact, (c) provides superior nuclear detail over standard histological sections and (d) may be an untapped research resource for the investigation of normal and pathological processes because of its ability to look at the placenta in a novel way and through its potential for both ex vivo and in vivo placental sampling.

Unusual location of a central neurocytoma: Particularities of a rare brain tumor

Introduction: Central neurocytoma is a typically rare intraventricular glioneuronal tumor in young adults. Clinically, it is revealed by symptomatology of intracranial hypertension. Intra-parenchymal, extraventricular, and intramedullary localizations are exceptional. Central neurocytoma is considered a benign tumor that can recur locally, but craniospinal dissemination remains as an exception. Case Report: We report a case in a 35-year-old male subject who consulted for quadriparesis and genito-sphincter dysfunctions. The clinical examination noted a syndrome of intracranial hypertension, a kinetic and static cerebellar syndrome, spastic tetraplegia with crural predominance, and anesthesia going up to the sixth thoracic vertebra (T6). A magnetic resonance imaging (MRI) showed a heterogeneous left cerebellar lesion. This lesion was excised, and the sample was freshly sent to the pathological anatomy laboratory. Extemporaneous cytology using the Smear technique was performed and was consistent with a hemangioblastoma. The standard histological examination also revealed the diagnosis of hemangioblastoma, and an additional immunohistochemical (IHC) study was performed for better diagnostic accuracy. The study was conducted in a North African laboratory and was conclusive of a neurocytoma. The evolution was marked by cerebellar tumor regrowth at five months, then the death of the patient at eight (08) months after surgery. Conclusion: Central neurocytoma is a rare tumor that can be confused with several other tumors of the central nervous system. Immunohistochemistry is essential for diagnostic confirmation.

A Pilot Validation Study Comparing Fluorescence-Imitating Brightfield Imaging, A Slide-Free Imaging Method, With Standard Formalin-Fixed, Paraffin-Embedded Hematoxylin-Eosin-Stained Tissue Section Histology for Primary Surgical Pathology Diagnosis.

Digital pathology using whole slide images has been recently approved to support primary diagnosis in clinical surgical pathology practices. Here we describe a novel imaging method, fluorescence-imitating brightfield imaging, that can capture the surface of fresh tissue without requiring prior fixation, paraffin embedding, tissue sectioning, or staining. To compare the ability of pathologists to evaluate direct-to-digital images with standard pathology preparations. One hundred surgical pathology samples were obtained. Samples were first digitally imaged, then processed for standard histologic examination on 4-μm hematoxylin-eosin-stained sections and digitally scanned. The resulting digital images from both digital and standard scan sets were viewed by each of 4 reading pathologists. The data set consisted of 100 reference diagnoses and 800 study pathologist reads. Each study read was compared to the reference diagnosis, and also compared to that reader's diagnosis across both modalities. The overall agreement rate, across 800 reads, was 97.9%. This consisted of 400 digital reads at 97.0% versus reference and 400 standard reads versus reference at 98.8%. Minor discordances (defined as alternative diagnoses without clinical treatment or outcome implications) were 6.1% overall, 7.2% for digital, and 5.0% for standard. Pathologists can provide accurate diagnoses from fluorescence-imitating brightfield imaging slide-free images. Concordance and discordance rates are similar to published rates for comparisons of whole slide imaging to standard light microscopy of glass slides for primary diagnosis. It may be possible, therefore, to develop a slide-free, nondestructive approach for primary pathology diagnosis.

Role of Kraevsky cells in forensic histological diagnostics

The study objective is to establish the role of light hepatocytes (Kraevsky cells) for forensic histological diagnosis of glycogen depletion in the liver due to various conditions, including shock. A standard histological examination of liver samples of persons who died from various causes was performed; routine staining with hematoxylin and eosin and additional staining by the PAS method was used. Histotopographic comparison of cross sections was performed to clarify the localization of light hepatocytes with different PAS staining results. It was shown that light hepatocytes have different morphological features, localization, and results of PAS staining, regardless of the cause of death, and are not related to glycogen depletion in them. Light hepatocytes (Kraevski cells) cannot be used as a sign of glycogen depletion and/or shock.

The clinical significance of the features of the morphological study and immunohistochemical determination of pancytokeratin in the lymph nodes of the central zone in papillary thyroid cancer

Introduction. papillary thyroid cancer is increasingly being detected at early stages when regional and distant metastases are absent per clinical examination. However, lymph nodes of the central zone can carry hidden metastases. frequency of such metastases is 22.3–46.7 %. maximally accurate identification of hidden metastases after lymph node dissection remains an important problem.Aim. тo evaluate the effect of histological examination characteristics and immunohistochemical measurement of pancytokeratin level on frequency of detection of papillary cancer hidden metastases in regional lymph nodes of the central zone in patients with clinical stage N0 papillary cancer.Materials and methods. The main group included 50 patients with stage ст1–2N0М0 primary papillary thyroid cancer. Dissected central lymph nodes of the patients prior to formalin fixation were extracted from the sample and inserted in individual paraffin blocks. Apart from standard histological examination, pancytokeratin level was measured immunohistochemically in the lymph nodes. The control group consisted of 200 patients for whom dissected central cell tissue was sectioned into blocks after formalin fixation. The number of lymph nodes in the dissected sample was measured by a pathomorphologist.Results. In the main group, the number of lymph nodes in the sample varied between 6 and 37 with mean of 20.7 ± 6.8; in the control group the number was lower: 3–25, mean 9.8 ± 5.1 (р = 0.000). In the main group, hidden metastases were detected more frequently than in the control group: in 30 (60 %) and 68 (34 %) cases, respectively (р = 0.001). In 20 (40 %) patients, immunohistochemical examination showed new metastases. use of this method allowed to detect 1 to 7 additional metastases (mean 2.4 ± 1.5 lymph node lesions).Conclusion. use of targeted dissection with extraction of lymph nodes led to significant increase in their numbers in the samples, and immunohistochemical examination allowed to detect a large number of hidden metastases in the central lymph nodes.

Technical note: A comparison between rehydrating solutions in the pretreatment of mummified and corified skin for forensic microscopic examination

Microscopic examination of mummified or corified skin may be of extreme importance for forensic purposes. However, standard histological samples in these cases are low-end, and preparation is burdened by several problems and so are diagnostic results: an improvement of these types of specimens is therefore advantageous. This study aims to identify the best performing rehydration solution among a fabric softener, a body lotion, and Sandison’s rehydrating solution. Samples of skin undergoing mummification or corification were collected from 25 corpses and each sample was divided into 4 fragments: one of these fragments was directly fixated in 4% formalin, one was previously treated with a tissue softener, another one was previously treated with a body lotion, and the last one was treated with Sandison’s solution. After 72 h, the pretreated samples were post-fixated in 4% formalin and then prepared for standard histological examination staining the histological slides with hematoxylin-eosin and Masson’s trichrome. At the microscopic examination, samples directly fixated in formalin were characterized by usual marked structural alterations and altered stainability, typical of such dry tissues. Vice versa, those previously treated appeared to be better-preserved even though with different improvement levels: body lotion made a medium–low-grade restoration of the tissues, and fabric softener a high-grade restoration, while Sandison’s rehydrating solution produced an optimal grade restoration. Sandison’s rehydrating solution was confirmed to be the best rehydrating substance for mummified and corified skin. Fabric softener could be, however, considered a valid substitute, being productive of high-grade microscopic yield.

Diagnostic Accuracy of Digital Staining ex vivo Confocal Microscopy for Diagnosing and Subtyping Basal Cell Carcinoma in Fresh Pretherapeutic Punch Biopsies: A Monocentric Prospective Study

Background: Ex vivo confocal microscopy using fusion mode and digital staining (EVCM) scans unfixed fresh tissue and produces rapidly digitally stained images of very similar quality to classical pathology. We investigated whether EVCM could represent an alternative to the standard histological examination of the pretherapeutic basal cell carcinoma (BCC) punch biopsies. Objectives: The objective of the study was to assess diagnostic accuracy of EVCM versus traditional histopathological examination for diagnosing and subtyping clinically suspicious lesions of BCC in 3-mm fresh and nonfixed punch biopsies. Methods: In this prospective monocentric observational study, patients with clinically suspected BCC were consecutively enrolled. Punch biopsies were imaged using EVCM and subsequently processed for standard histologic examination (gold standard). EVCM images were examined by a dermatopathologist blinded to clinical aspect of the lesion and histopathological results. Concordance between the EVCM and histology analysis was calculated with Cohen’s kappa (κ) statistic. Results: Sixty-six patients were recruited, and 106 biopsies were analyzed. EVCM correctly diagnosed 70/73 BCCs and 31/33 non-BCC lesions, corresponding to a sensitivity of 96% and a specificity of 94% (positive predictive value = 97%, negative predictive value = 91%). The EVCM assessment led to over-staging and under-staging of BCC subtypes in 5% and 11% of cases, respectively. It led to over-staging and under-staging of BCC depths in 5% and 15%, respectively. The kappa coefficient for concordance was 0.78 (95% confidence interval [CI]: 0.69–0.88) when considering BCC subtypes and 0.81 (95% CI: 0.72–0.90) when considering BCC depths. Conclusions: These results render EVCM as a promising option for “real-time” pretreatment evaluation of clinically suspected BCC lesions. Further larger randomized studies are needed to assess the efficiency of EVCM versus standard care in patients with clinically suspected BCC.

AUTOFLUORESCENCE SPECTRA OF A SPECIMEN OF NON-KERATINIZING LARYNGEAL SQUAMOUS CELL CARCINOMA IN ACCORDANCE WITH A HISTOLOGICAL STUDY

Abstract. The prevalence of oncological diseases of ear, nose and throat organs tends to increase in recent years. This paper describes a method for the early diagnosis of diseases of the pharynx and larynx, accompanied by epithelial dysplasia using autofluorescence spectroscopy. Fluorescence spectroscopy is a non-invasive, highly sensitive, versatile and relatively easy-to-use method that allows combined diagnostics and the ability to work in real time without tissue trauma. Its use in the diagnosis of various diseases is due to the fact that during the development of pathologies, biochemical changes occur in the tissue, and, consequently, the relative content of the main fluorophores and chromophores changes, the morphological and histological parameters of the tissue change and, as a result, their contribution to the overall observed fluorescence spectrum fabrics. Fluorescence was started using an LGI-505 nitrogen laser (excitation wavelength 337 nm, pulse repetition rate up to 1 kHz, pulse duration 6–10 ns). Registration of autofluorescence spectra carried out by usage a spectrum analyzer with fiber optic input LESA-7med. For the study, we used a sapecimen of the larynx obtained during laryngectomy wich previously histologically confirmed as laryngeal cancer. Biopsy performed in limited areas of the mucosa determined by the autofluorescence spectra analysis. Then a standard histological examination of biopsy samples were performed. A direct correlation was found between the results of autofluorescence spectroscopy and histological examination of identical localization areas of the object under study. Thus, squamous cell carcinoma is characterized, along with a general decrease in fluorescence intensity, by the shift of the main maximum of the spectrum in the region of 435-455 nm. The low content of NADH in tumor cells, as well as dysproteinemia developing in malignant diseases, are the main reason for the decrease in the intensity of the autofluorescence spectrum.

Assessment of Extramammary Paget Disease by Two-Photon Microscopy.

Two-photon microscopy techniques are non-linear optical imaging methods which are gaining momentum in the investigation of fixed tissue sections, fresh tissue or even for in vivo experiments. Two-photon excited fluorescence and second harmonic generation are two non-linear optical contrast mechanisms which can be simultaneously used for offering complementary information on the tissue architecture. While the former can originate from endogenous autofluorescence sources (e.g., NADH, FAD, elastin, keratin, lipofuscins, or melanin), or exogenous eosin, the latter is generated in fibrillar structures within living organisms (e.g., collagen and myosin). Here we test the ability of both these contrast mechanisms to highlight features of the extramammary Paget disease on fixed tissue sections prepared for standard histological examination using immunohistochemical markers and hematoxylin and eosin staining. We also demonstrate the label-free abilities of both imaging techniques to highlight histological features on unstained fixed tissue sections. The study demonstrated that two-photon microscopy can detect specific cellular features of the extramammary Paget disease in good correlation with histopathological results.

Morphologically, immunohistochemically and PCR proven lymphocytic viral peri-, endo-, myocarditis in patients with fatal COVID-19

BackgroundDespite a reported cardiac injury in patients with new coronavirus infection, the possibility and specifics of genuine viral myocarditis in COVID-19 remains not fully clear.PurposeTo study the presence of SARS-CoV-2 in the myocardium and the morphological properties of myocarditis in patients with severe coronavirus infection (COVID-19).MethodsAutopsy data of eight elderly patients (75.6 ± 7.4 years), four male and four female, with severe new coronavirus infection were studied. The lifetime diagnosis of COVID-19 is based on a positive result of the PCR study. The inclusion criterion was the presence of morphological signs of myocarditis according to the Dallas criteria. A standard histological examination included staining by hematoxylin and eosin, toluidin blue and Van Gieson. An immunohistochemical study was performed using antibodies to CD3, CD 68, CD20, perforin, toll-like receptor (TLR) types 4 and 9. PCR in real-time was performed to determine the viral RNA in the myocardium.ResultsAll patients had severe bilateral viral pneumonia. In all cases, myocarditis was not clinically diagnosed. Morphological examination of the heart found signs of active lymphocytic myocarditis. PCR identified the SARS-Cov2 RNA in all cases. There were also signs of destructive coronaritis in all cases, thrombovasculitis, lymphocytic pericarditis (in 3 cases) and endocarditis (in 2 cases). The absence of neutrophils confirms the aseptic nature of inflammation. An immunohistochemical study showed the CD3-positive T lymphocytes in the infiltrates. Increased expression of TLR type 4 and less 9 was also detected.ConclusionMorphological and immunohistochemical evidence of myocarditis in COVID-19 was presented. Lymphocytic infiltrations and positive PCR confirm the viral nature of inflammation. Myocarditis in COVID-19 is also characterized by coronaritis with microvascular thrombosis and associated with lymphocytic endo- and pericarditis.

Artificial intelligence and deep learning to map immune cell types in inflamed human tissue

Biopsies of inflammatory tissue contain a complex network of interacting cells, orchestrating the immune or autoimmune response. While standard histological examination can identify relationships, it is clear that a great amount of data on each slide is not quantitated or categorized in standard microscopic examinations. To deal with the huge amount of data present in biopsy tissue in an unbiased and comprehensive way, we have developed a deep learning algorithm to identify immune cells in biopsies of inflammatory lesions. We focused on T follicular helper (Tfh) cell subsets and B cells in dermatomyositis biopsy images. We achieved strong performance on detection and classification of cells, including the rare Tfh cell subsets present in the tissue. This algorithm could be used to perform distance mapping between cell types in tissue, and could be easily adapted to other disease states.

Superficial basal cell carcinoma, think deeper: Step sectioning of skin biopsy specimens yields 14% more aggressive subtypes.

IntroductionBecause of different therapeutic regimens for superficial and non-superficial basal cell carcinomas (BCCs), accurate histopathological examination of a punch biopsy to determine its subtype is essential. The aim of the current study was to evaluate the additional yield of a more extensive step-section method to that of a standard histologic examination at 4 levels.Material and methodsData for this prospective study was obtained from the Pathology department of a Dutch tertiary hospital. Biopsy specimens of subsequent patients from March 2019 to June 2020 were sectioned to 8-levels instead of the regular 4-levels. Only patients with a superficial BCC subtype in the first 4-levels of sectioning were included (n = 100). After 8-level sectioning, it was recorded in which level (5–8) a more aggressive BCC component was found (i.e. nodular, infiltrative, or micronodular). Patients were followed-up to evaluate further treatment, and in case of excision, the excision specimen was reviewed to determine the BCC subtype. A logistic regression was performed to assess characteristics associated with a more aggressive BCC component in levels 5–8.ResultsIn 14 patients (14%) a more aggressive component was found in levels 5–8, all with a nodular component. Thirteen of these patients underwent excision, confirming a more aggressive BCC subtype. Of the 86 patients that had no deeper BCC component in levels 5–8, 26 (30.2%) underwent excision; In 4 patients no residual BCC was found, in 15 patients superficial BCC, and in 7 a more aggressive BCC subtype (1 nodular and 6 a combination of superficial/nodular/infiltrative). In multivariable analysis, head&neck localization was associated with finding a more aggressive BCC subtype in levels 5–8 (OR 6.41 (95%CI 1.56–26.30), p = 0.01)).ConclusionsMore extensive sectioning of superficial BCC biopsy specimens, especially in the head&neck area, leads to a more accurate BCC subtype diagnosis requiring different clinical management strategies.

Histotopography of haemorrhagic infiltration in the hanging cutaneous furrow: Where to look for haemorrhagic infiltration in hanging.

The forensic evidence of hanging is based on the autopsy demonstration of the traces left by the noose or the ligature on the neck, as well as on the histological assessment of the hanging mark vitality. However, the specific topography of haemorrhagic infiltration in the context of the cervical damaged tissues involved in hanging is not known. We carried out an extensive microscopic examination to identify if haemorrhagic infiltration in hanging appeared in specific topographic locations that could have been considered as elective. From 102 victims of suicidal hanging, a skin fragment was sampled from the maximum compression area of the sulcus, including the skin portions immediately above and below it. The sampling was also extended to collect the subcutaneous adipose tissue and part of the striated muscle. A standard histological examination was performed on all the samples, and hematoxylin and Eosin, Weigert's resorcin-fuchsin and Goldner's Masson trichrome staining were performed. In all the cases assessed, the microscopic examination allowed the detection of haemorrhagic infiltration mainly in the deepest areas under the sulcus and especially in the deep dermis and at the transition point between the dermis and the subcutaneous adipose tissue, as well as in the context of its supporting connective tissue. Such areas could therefore be considered as regions in which the presence of haemorrhagic infiltration is more likely to be demonstrated. Accordingly, we recommend performing sampling similar to ours and focusing the search for haemorrhagic infiltration as suggested.

Experimental Modeling of the Ultra-Necrotic Stage of Critical Ischemia 
 of the Lower Limbs

Objective. So far, no experimental model of the necrotic ulcerative stage of critical limb ischemia has been created. Aim. Development of a method for modeling the necrotic ulcerative stage of critical lower limb ischemia.Materials and methods. The experimental study was carried out on 120 rats - males of the "Wistar" line, divided into 4 groups of 30 animals each. In the first group (intact), the level of microcirculation in the muscles of the hind limb was determined. In the second (sham-operated) group, ischemia was not simulated, but the neurovascular bundle of the thigh was isolated and the wound was sutured. In the third group (comparison), the modeling of chronic ischemia of the leg muscles was performed by removing the femoral, popliteal, anterior and posterior tibial arteries. In the fourth (experimental) group, to simulate chronic critical ischemia of the leg muscles, the femoral, popliteal, anterior and posterior tibial arteries and veins were removed, and the peripheral nerve was damaged.In the postoperative period, the dynamics of symptoms of limb ischemia was monitored. Animals were taken out of the experiment, 10 animals in each group by an overdose of anesthesia on the 10th, 21st and 28th days. Before hatching, the level of microcirculation was determined using laser Doppler flowmetry. To study the morphological picture at the same time in the third and fourth groups, the ischemic gastrocnemius muscle of the leg was excised and a standard histological examination was performed.Results. Trophic disorders in animals of the third group developed in 13.3%, and in the experimental group - in 100%. In the experimental group, the level of microcirculation at all periods was significantly less than in the comparison group: on the 10th and 21st by 1.3 times, on the 28th day by 1.2 times, and the area of ​​muscle necrosis was greater by the same days by 21.2%, by 8.2% and by 6.8%.Conclusion. The developed experimental model corresponds to the necrotic ulcerative stage of critical ischemia in humans.

Targeted use of intraoperative frozen-section analysis lowers the frequency of completion thyroidectomy

BackgroundThe impact of intraoperative frozen section (iFS) analysis on the frequency of completion thyroidectomy for the management of thyroid carcinoma is controversial. Although specialized endocrine centres have published their respective results, there are insufficient data from primary and secondary healthcare levels. The aim of this study was to analyse the utility of iFS analysis.MethodsIn the Prospective Evaluation Study Thyroid Surgery (PETS) 2 study, 22 011 operations for benign and malignant thyroid disease were registered prospectively in 68 European hospitals from 1 July 2010 to 31 December 2012. Group 1 consisted of 569 patients from University Medical Centre (UMC) Mainz, and group 2 comprised 21 442 patients from other PETS 2 participating hospitals. UMC Mainz exercised targeted but liberal use of iFS analysis for suspected malignant nodules. iFS analysis was compared with standard histological examination regarding the correct distinction between benign and malignant disease. The percentage of completion thyroidectomies was assessed for the participating hospitals.ResultsiFS analysis was performed in 35.70 per cent of patients in group 1 versus 21.80 per cent of those in group 2 (risk ratio (RR) 1.6, 95 per cent c.i. 1.5 to 1.8; P < 0.001). Sensitivity of iFS analysis was 75.0 per cent in group 1 versus 63.50 per cent in group 2 (RR 1.2, 1.2 to 1.3; P = 0.040). Completion surgery was necessary in 8.10 per cent of patients in group 1 versus 20.8 per cent of those in group 2 (RR 0.4, 0.2 to 0.7; P = 0.001).ConclusioniFS analysis is a useful tool in determining the appropriate surgical management of thyroid disease. Targeted use of iFS was associated with a significantly higher sensitivity for the detection of malignancy, and with a significantly reduced necessity for completion surgery.

Морфологические варианты гравидарной трансформации эндометрия при неразвивающейся беременности после экстракорпорального оплодотворения

Introduction. High prevalence of early pregnancy loss regardless of methods of conception still remains a relevant problem. One of the ways to identify the etiology of pregnancy loss is a histological examination of the products of conception (POC), decidualization in particular, which often remains underestimated in clinical practice. The aim of the study was to evaluate the decidualization of the endometrium in the retained POC after IVF. Materials and methods.Histological and immunohistochemical analysis of decidualized endometrium was carried out on 153 samples of retained POC around 5–8 weeks of gestation after IVF and on 15 samples of POC after artificial abortion. The POC was fixed in a 10% neutral formaldehyde solution, and a standard histological examination was performed. The immunohistochemical study was carried out to evaluate the expression of estrogen and progesterone receptors in the endometrial glands and stromal cells of the decidualized endometrium compact layer. Results. We verified 5 types of decidual endometrial stromal and glandular transformation of a missed abor-tion material after IVF. The morphological features of decidaulized endometrium in missed abortion after IVF are characterized by a complete and impaired stromal transformation with various types of glandular transformation and disynchronous expression of estrogen and progesterone receptors. Conclusion.We described the morphological variants of the decidual endometrial transformation in POC after IVF, which serve a new direction to the understanding of endometrial factor as one of the main determinants of recurrent implantation and reproductive failure often remaining out of the specialist’s focus. Keywords: missed abortion, complete and incomplete gravidar transformation of the endometrium, estrogen and progesterone receptors, IVF

The Value of Calretinin in the Diagnosis of Hirschsprung&amp;apos;s Disease in Dakar

<i>Introduction</i>: Hirschsprung's disease is a congenital anomaly characterised by a rarefaction or absence of ganglion cells associated with schwannian hyperplasia or hypertrophy involving the colon or a segment of the colon. Its diagnosis is based on clinical, radiological and histological evidence. Haematoxylin-eosin examination is used in cases of suspected Hirschsprung's disease. However, in some situations it is insufficient to confirm or refute the diagnosis. Immunohistochemical and histo-enzymatic techniques can be used to demonstrate aganglionosis. The aim of our study is to evaluate the interest of calretinin in the diagnosis of Hirschsprung's disease in Dakar. <i>Materials and methods</i>: This was a retrospective and descriptive study from 1 January 2018 to 31 August 2019. It was conducted at the pathology laboratory of Idrissa Pouye General Hospital where 51 paraffin blocks from suspected Hirschsprung's Disease cases were included. All blocks were immunohistochemically studied with anti-calretinin antibody. The diagnosis of Hirschsprung's disease on standard histological examination was based on the absence of lymph node cells in the submucosal and myenteric plexuses. Calretinin immunoreactivity was shown by nuclear and cytoplasmic labelling of ganglion cells and nerve nets. In Hirschsprung's disease, there is an absence of labelling of the nerve plexuses. <i>Results</i>: The majority of patients (73.5%) were aged 2 years or older with a mean age of 3.4 years. The sex ratio was 2. Biopsies constituted 56.86% of the specimens and surgical specimens 43.14%. Concordance between haematoxylin-eosin examination and calretinin immunohistochemistry was observed in 47 cases (92.15%) and discordance in 4 cases (7.15%). The sensitivity of calretinin was 93.75% and the specificity 89.47%. The kappa index was 0.92. The recto-sigmoid form was the most frequent topographic form observed in 83.3% of patients. <i>Conclusion</i>: Morphological examination with haematoxylin-eosin remains a good diagnostic method for Hirschsprung's disease. Calretinin immunohistochemistry is necessary in equivocal cases, in neonates and infants and in case of superficial biopsies.

Helicobacter pylori-associated gastritis phenotypes in a South-Western Nigerian population

Background: The long-term outcomes of Helicobacter pylori infection ultimately depend on the distribution and extent of gastritis. Three main types of gastritis phenotypes which include pangastritis, antral-predominant gastritis, and corpus-predominant gastritis have been observed. Corpus predominant gastritis is more common in the parts of Asia with a high prevalence of gastric adenocarcinoma. This study was conducted to determine the prevalence of H. pylori-associated gastritis phenotypes in a South-western Nigerian population, a sub-Saharan African population. Materials and Methods: The study was a cross-sectional prospective study which included 267 dyspeptic patients who had esophagogastroduodenoscopy at a tertiary health institution in South-west Nigeria. Five gastric biopsies were taken from each subject, and standard histologic examination was performed on the specimens. Both biopsy sampling and histologic examination followed the Updated Sydney System recommendations. The data generated were statistically analyzed. Continuous variables were presented as means ± standard deviation, whereas associations between the categorical variables were determined by the Pearson Chi-square test or Fisher's exact test. P ≤ 0.05 was considered statistically significant. Results: The mean age was 46.1 (±15.4) years while the age range was 16–84 years. There were 160 (59.9%) female and 107 (40.1%) male participants. All participants had histological gastritis, whereas 79 (29.6%) were positive for H. pylori by histology. H. pylori infection was significantly associated with peptic ulcer disease. Among those who had H. pylori infection, 51 (64.6%) had pangastritis, 26 (32.9%) had antral-predominant gastritis, while 2 (2.5%) had corpus-predominant gastritis. The relationship between H. pylori infection and the gastritis phenotypes showed no statistical significance. Conclusion: The prevalence of corpus predominant gastritis was low in our study population.

Features of oral squamous cell carcinoma in ex vivo fluorescence confocal microscopy.

Real-time microscopic imaging of freshly excised oral squamous cell carcinomas (OSCCs) would be potentially supportive in rapid recognition of oral malignancy and an optimal and time-saving management of patients' surgical treatment. The aim of this study was to examine oral squamous cell cancer tissue in regards to the commonly known and well-described histomorphologic criteria for the diagnosis of OSCC in ex vivo confocal fluorescent microscopy and to analyze its correlation with grade of differentiation and level of invasiveness. Ex vivo confocal laser scanning microscopy (CLSM) images of 38 OSCCs were evaluated immediately after excision for presence or absence of various cytological and architectural features based on the histopathological background. Next, these features were compared to the grade of differentiation as elaborated via gold standard histologic examination. Of 38 invasive OSCCs, 14 were well differentiated, while three moderately and 19 were poorly differentiated. The presence of the commonly known cytologic and histopathologic criteria for the diagnosis of oral squamous cell carcinoma such as the destruction of the basal cell membrane, cellular and nuclear pleomorphism, anisocytosis, intraepithelial keratinization, nuclear hyperchromasia, atypical mitotic figures as well as the presence of necrosis, and mixed inflammation could be observed in ex vivo fluorescence confocal microscopy (FCM). In ex vivo fluorescence confocal microscopy pictures, cellular pleomorphism and anisocytosis were observed more often in poorly differentiated OSCCs. Intraepithelial keratinization was associated with well differentiated and moderately differentiated OSCCs. The results demonstrate the high potential of ex vivo fluorescence confocal microscopy in fresh tissue for rapid real-time diagnosis of OSCC.