Papanicolaou Stain Research Articles

Comparative Study of Gynaecological and Non-Gynaecological Samples Received in Igbinedi on University Teaching Hospital

Background: Gynaecological and non-gynaecological samples are two main types of cytological sample. The Gynaecological samples are usually Pap (cervical) smears, Vagina smear, Endocervical aspiration, Endometrial aspiration etc. Non-gynaecological samples fall into two categories, exfoliative and fine needle aspiration samples. The sample size of this study was 32 and was determined using the method of Naing, Winn, and Rusli, (2006).Aim: This study was aimed at comparing the efficacy of the gynaecological sample and non-gynaecological sample in a variety of parameters including cellular morphology, specimen preparation, screening time, and impact on the final result. Materials and methods: Gynaecological sample was obtained by introducing a spatula into the vagina with the aid of speculum. The cellular material obtained was rapidly but gently smeared on a clean glass slide and the smear made was fixed while it is wet for Papanicolaou staining and was cleared, mounted and studied microscopically. Non-Gynaecological sample – sputum was collected with a deep cough from the lungs early in the morning before eating and deposited directly into a preservative container. Air-dried smears were fixed in methanol for 15 minutes and rinsed in buffer. The smear was stained with May-Grunwald for 5 minutes and stained again with Giemsa for 10 minutes. The smear was differentiated in buffer and allowed to air dried, cleared in xylene and mounted with DPX.Results: This present study revealed that cervical casesoccurred more in younger age (30-39) when compared to non gynaecological cases (sputum) which occurred more in older adults (age group 50-59).Conclusion:Treating all cervical pre-cancers can prevent almost all cervical cancers. Cervical smear showed predominately parabasal and intermediate cell, esinophilic cells with increased nuclear/ cytoplasmic ratio and eccentric cells. However, the sputum cytology show basically cells with increased nuclear/ cytoplasmic ratio.

Reference standards for gene fusion molecular assays on cytological samples: an international validation study

AimsGene fusions assays are key for personalised treatments of advanced human cancers. Their implementation on cytological material requires a preliminary validation that may make use of cell line slides mimicking...

English

BACKGROUND The axilla is a triangular space between the lateral wall of thorax and the upper part of arm. The component of axilla can give rise to different diseases ranging from developmental and reactive to neoplastic. Various mesenchymal tissues present in the axilla can give rise to different lesions. Fine needle aspiration cytology (FNAC) is a safe, quick, easy, and a cheap diagnostic tool to decipher the benign or the malignant nature of the swelling. Imaging techniques and FNAC, both used individually or coupled, improve the accuracy of diagnosis and help in avoiding unnecessary invasive procedures. The most common lymph node lesion in axilla is benign reactive lymph node hyperplasia. It is caused by inflammation of the lymph nodes or the adjacent organs, such as the breast or lung. METHODS The present study is a retrospective review of swellings aspirated from axillary region. FNAC was performed following strict aseptic precautions. Inadequate aspirates were excluded from the study. The aspirated sample was blown on clean glass slides and smears were made using squash preparation. Smears were fixed using air dried and alcohol fixation method. Smears were stained with Giemsa stain, and the alcohol - fixed smears were stained with haematoxylin and eosin and Papanicolaou stains. Cytological smears were evaluated for adequacy, cellularity, type of cells, arrangement of cells and nuclear as well as cytoplasmic features. RESULTS A total of 191 axillary swelling were aspirated during the study period. Out of which 148 (77.5 %) cases were females and 43 (22.5 %) cases were males. Most common lesion was lipoma followed by axillary tail of breast. CONCLUSIONS Various pathological processes may affect the axilla. There is overlap in the imaging findings of these diseases. FNAC is a diagnostic tool which is required for conformation of diagnosis of axillary lesions. KEYWORDS Axillary Swelling, Fine Needle Aspiration, Lipoma

P–257 An unknown cause lead to polyspermy in IVF cycles and 0PN zygotes in ICSI cycles in male patient

Abstract Study question The patient sperm has normal morphology and motility, which paternal factors cause the abnormal fertilization in IVF/ICSI and what is the underlying mechanism? Summary answer A genetic mutation of BEX1 and decreased PLC-zeta has been found in patient, which may provide novel insights of polyspermy and pronucleus formation during fertilization. What is known already In mammals, pronucleus formation, a landmark event for fertilization, is critical for embryonic development. Abnormal fertilization refers to the abnormal number of pronucleus and polar bodies in zygotes during in vitro fertilization, with an incidence of 5–15%, among which the incidence of polyspermy and 0PN is about 2–10% and 30%. However, the mechanisms underlying pronucleus formation still unclear. More research has focused on oocyte activation, while paternal relevant abnormal fertilizations have been rarely established. The mechanism of how sperm and/or substances carried by sperm influence the physiological process of fertilization is also unclear. Study design, size, duration In our study, we first work on the preliminary observation and analysis of sperm morphology, structure and sperm chromosome number, and then made further analysis at the genetic level to find out the cause of this particular phenotype in this patient. We use of zone-free golden hamster ova test the fertilizing capacity and rescue the pronucleus formation with SrCl2. Participants/materials, setting, methods The patient, golden hamster, Papanicolaou stain, scanning electron microscope (SEM), Transmission Electron Microscope (TEM), Fluorescence in situ hybridization (FISH), Whole Exome Sequencing (WES), IVF, ICSI, Assisted Oocyte Activation (AOA). Main results and the role of chance During 2016–2018, they did 4 cycle assistant reproduction technology. Cycle1, conventional IVF(C-IVF), 9 MII oocytes, 9 3PN zygotes; Cycle2, ICSI, 10 MII oocytes, 10 0PN zygotes; Cycle3, donor-oocytes C-IVF, 6 MII oocytes, 6 3PN zygotes, and the donor did C-IVF get normal zygotes and embryos; Cycle4, donor-sperm C-IVF, 7 MII oocytes, 4 2PN zygotes, 3 useable embryos. Remarkably, clinical examination about male shows normal sperm semen parameters. Papanicolaou stain and SEM shows that the sperm of the patient has normal morphology. The TEM data shows that the spermatozoa with normal head morphology and intact 9 + 2 sperm flagella structure. In the sperm FISH analysis, Chromosome ploidy is haploid. We performed WES on the male, after exclusion of frequent variants and application of technical and biological filters, two homozygous missense mutations were identified in BEX1 (c.191G>A [p. W64X]), which has been few reports of male infertility. The western blot result show that the PLC-zeta was decreased in patient. After 10mM SrCl2 assisted oocyte activation, the zygote has the pronucleus formation in ICSI. Limitations, reasons for caution At present, we only observe sperm related factors (morphology, structure, chromosome number, genetic mutation). Next step is to detect the substances sperm carried (e.g. RNA-seq, proteomics). In this case, what is of great concern to us is the inconsistencies of the abnormal fertilization during the conventional IVF and ICSI cycles. Wider implications of the findings: Many studies of fertilization mechanism, the main focus is on the maternal cytoplasmic factors, such as the Ca 2+ release initiate the fast block of oocytes. There are few reports about abnormal fertilization due to sperm factors. Our case may offer new insights for the study of fertilization. Trial registration number Not applicable

Roflumilast improves resolution of sepsis-induced acute kidney injury by retarding late phase renal interstitial immune cells infiltration and leakage in urinary sediments.

Some evidence has demonstrated that both inflammation and immune cell dysregulation are coincident at late phase (post 24 h) of sepsis. The present study was designed to determine the pathological role of hyperinflammation and renal immune cells mobilization during late phase of sepsis induced acute kidney injury (S-AKI) and tests the pharmacological effects of PDE-4 inhibitor on these events. Sepsis was induced by cecal ligation puncture and renal function, oxidative-inflammatory stress biomarkers were assessed after 24 h. PDE-4 inhibitor was administered for 7 days prior to induction of S-AKI. Renal immune cells infiltration during sepsis was analyzed by H&E staining and papanicolaou staining method was used for detecting leukocytes and cast in urinary sediments, periodic acid schiff (PAS) staining was used for detection of brush border loss. AKI developed 24 h post sepsis insult as depicted by increase in serum creatinine, blood urea nitrogen (BUN), renal oxidative stress, and elevated inflammatory biomarkers levels. Moreover, septic rats displayed increased bacterial load, renal expression of phosphodiesterase-4B, 4D isoforms, enhanced vascular permeability, caspase-3 and myeloperoxidase activity, electrolyte imbalance, reduced Na+ K+ ATPase activity, declined cAMP levels, increased interstitial leukocyte infiltration, and leakage in urinary sediments along with histological alterations. Pre-treatment with roflumilast at high dose completely prevented the various AKI associated manifestations in septic rats. Renal hyper-inflammation and leukocyte infiltration was detected in late phase of S-AKI. Roflumilast pre-treatment resolved sepsis induced renal dysfunction and histological damage by suppressing late phase renal immune cells invasion and anti-inflammatory effects mediated by up-regulation of renal cAMP levels.

Cytomorphological Spectrum of Salivary Gland Lesions by Fine Needle Aspiration Cytology

Introduction: Salivary gland lesions encompass a baffling subset of lesions with overlapping morphologies on cytomorphology, due to their heterogenous histologies and rare enough to sometimes bewilder even the most knowledgeable cytopathologists. The technique of fine needle aspiration (FNA) cytology in the evaluation of salivary gland lesion is simple to perform, saves time, reliable, safe and inexpensive. FNA is highly accurate when used in proper clinical setting and supported by appropriate clinical and relevant diagnostic data. Rapidity of obtaining a pathological diagnosis allows more intelligent therapeutic approach. Materials and Methods: It was a prospective study carried out in the cytology section of pathology department for a period of two years using fine needle aspiration as a diagnostic tool as per standard protocol. The clinical and radiological data were obtained from the patient’s case papers and collaboration with the operating surgeon, the lesions were analysed on cytology. Haematoxylin & Eosin, Papanicolaou stain and Giemsa were used as standard stains. Results: A total of 66 cases of salivary gland lesions were aspirated for a period of two years. Majority of cases were in the age group of 21-30 years with 36 male patients (55%) and 30 female patients (45%). About 44(66.7%), 19(28.8%), 1(1.5%) and 2(3.0%) cases occurred in parotid gland, submandibular gland, sublingual gland and minor salivary gland respectively. There were 21(32%) cases of non-neoplastic lesions and 45(68%) cases of neoplastic lesions. Histopathological specimens were available in 11 out of 66 cases and showed correlation with cytological findings. Conclusion: Fine needle aspiration cytology of salivary gland lesions has high diagnostic accuracy, helps in appropriate therapeutic management and is useful as a diagnostic procedure because of the availability of earlier diagnosis in comparison with the histopathology. Keywords: Salivary gland lesions, fine needle aspiration cytology, neoplastic, non-neoplastic.

Constitutive Cytomorphologic Features of Medullary Thyroid Carcinoma Using Different Staining Methods.

(1) Background: Accurate preoperative identification of medullary thyroid carcinoma (MTC) is challenging due to a spectrum of cytomorphologic features. However, there is a scarcity of studies describing the cytomorphologic features as seen on fine-needle aspiration (FNA) smears prepared using different staining methods. (2) Methods: We performed a retrospective study on MTC cases with available FNA slides from 13 hospitals distributed across 8 Asia-Pacific countries. The differences in the constitutive cytomorphologic features of MTC with each cytopreparatory method were recorded. A comparative analysis of cytologic characteristics was carried out with appropriate statistical tests. (3) Results: Of a total of 167 MTC samples retrospectively recruited, 148 (88.6%) were interpreted as MTC/suspicious for MTC (S-MTC). The staining methods used were Papanicolaou, hematoxylin-eosin, and Romanowsky stains. Seven out of the eleven cytologic criteria can be readily recognized by all three cytopreparatory methods: high cellularity, cellular pleomorphism, plasmacytoid cells, round cells, dyshesive cells, salt-and-pepper chromatin, and binucleation or multinucleation. An accurate diagnosis was achieved in 125 (84.5%) of the 148 samples whose FNAs exhibited five or more atypical features. Conclusions: The present work is the first study on MTC to compare the morphological differences among the cytologic staining techniques. We investigated the constitutive features and the reliability of diagnostic parameters. A feasible scoring system based upon cytomorphologic data alone is proposed to achieve a high degree of diagnostic accuracy.

Significance of Reporting Cannon Balls in Diagnosing Bacterial Vaginosis Infection in PAP Smear

Bacterial vaginosis is the most common cause of vaginitis. It is caused by depopulation of lactobacilli from the normal vaginal flora and overgrowth of Gardnerella vaginalis and other anaerobic species. Pap smear being a screening and diagnostic tool for malignancy, is equally important to diagnose infective pathology as well. This study tried to establish the significance of reporting the presence of cannon balls in Pap smears and whether their presence suggested bacterial vaginosis or not. This was a cross sectional study conducted at Nepal Medical College, Teaching Hospital (NMCTH), Kathmandu for duration of one year (January 2020 to December 2020). Pap smears were sent from gynaecology department and standard routine Pap stain was done in Pathology Department. Presence of bacterial vaginosis and cannon balls were noted. Total of 799 Pap smears were received. The mean age of bacterial vaginosis was 38.2 ± 9.6 years. The age group of 31-40 years (n=32, 39%) was most commonly affected. Bacterial vaginosis was seen in 81 pap smears out of 799 pap smears. The prevalence of bacterial vaginosis was 10.1%. Of the total of 799 cases of Pap smears, cannon balls were seen in 9.1% cases and among the bacterial vaginosis, cannon balls were seen in 48% cases. There was significant association between cannonballs and bacterial vaginosis (p < 0.0001). Thus, the present study re-enforces the fact that Pap smear is also very effective tool to diagnose bacterial vaginosis.

Cytomorphological Profile of Thyroid Lesions on Fine Needle Aspiration Cytology

Background and Objectives: Fine needle aspiration cytology (FNAC) of thyroid offers an alternative, as an immediate, preliminary procedure associated with low cost, little trauma and complications. Being superficial and easily accessible it is an ideal organ for FNAC. It is useful in diagnosis of inflammatory, infectious and neoplastic conditions. The aim of this study was to evaluate the cytomorphological profile of thyroid lesions on FNAC with application of Bethesda system for reporting and perform cyto-histopathology correlation whenever possible with regards to age and sex distribution. Methodology: This two-year prospective study was conducted in the Cytology section of Pathology Department & included 243 cases of thyroid gland enlargement. The detailed history of the patient including age, sex, presenting symptoms and duration of thyroid gland enlargement along with any other significant findings was note. FNAC was performed as per the protocol using a 23-gauge needle and smears stained using Haematoxylin & Eosin (H&E) and Papanicolaou stain, while air-dried smears were stained with Giemsa stain. Results: The study period included a total of 243 FNACs of thyroid enlargement. The age of patients ranged from 10 to 80 years with male to female ratio of 1.4. The diagnosis on FNAC was as follows Colloid Goitre (54.7 %), Lymphocytic Thyroiditis (9.54%), Follicular Neoplasm (7.05%), Hashimoto Thyroiditis (5.80 %), Benign Thyroid Lesions (3.31%) and Multinodular Goitre (2.90%). Conclusion: Thyroid fine needle aspiration cytology has proven to be a first line tool to evaluate the thyroid lesions because of its cost effectiveness and high patient acceptance. Fine needle aspiration cytology is highly successful in triaging patients with thyroid nodules into operative and non-operative groups and enables surgeons to take an early decision regarding mode of treatment. The Bethesda system for reporting of thyroid lesions aims at standardization of reports. It makes the cytology report unambiguous and clinically relevant. Keywords: Fine needle aspiration cytology, thyroid lesions.

Cytotoxicity, genotoxicity, and cellular metal accumulation caused by professionally applied fluoride products in patients with fixed orthodontic appliances: A randomized clinical trial

Corrosion of metal orthodontic appliances caused by professional fluoride products has been recently concerned. Therefore, the objective of this study was to evaluate the cytotoxic and genotoxic effect of these products on buccal mucosal cells from patients wearing fixed orthodontic appliances. A total of 44 patients, aged 12 to 35 years, who began orthodontic treatment with fixed appliances were included in this single-center, prospective, randomized clinical trial. Patients were randomly allocated into 4 parallel groups according to the type of professional fluoride treatment applied after placing the appliances: acidulated phosphate fluoride gel (APF); neutral fluoride gel (NGel); fluoride varnish (FVa); and without fluoride treatment (control). Buccal cells were collected before treatment (T1) and 3 months after appliance placement (T2). The cells were assayed for cell viability and underwent Papanicolaou staining. Cells with micronuclei and degenerative nuclear alterations were scored using a light microscope. Cell metal content was quantified by inductively coupled plasma-mass spectrometry. The data were analyzed with the Kruskal-Wallis test. The intracellular nickel content in the APF group significantly increased (P < 0.05), whereas that of the control, NGel, and FVa groups did not. The changes in chromium concentration in all groups were not significantly different compared with control. Use of APF resulted in a significantly higher decrease in cell viability and increase in morphologic signs of cell death compared with control (P < 0.05). The change in frequency of micronucleated cells was not significantly different from that in the control group. Applying APF gel on fixed orthodontic appliances increased the cell metal content and decreased cell viability; however, genotoxic effects were absent. FVa and NGel are suggested as the products of choice to use during orthodontic treatment.

Application of the Bethesda system of reporting for cervical cytology to evaluate human papilloma virus induced changes in oral leukoplakia, oral squamous cell carcinoma, and oropharyngeal squamous cell carcinoma: A cytomorphological and genetic study.

Human papilloma virus (HPV) has a well-established carcinogenic role in certain head and neck cancers. These HPV associated cancers possess unique clinicopathological behavior and exhibits better prognosis than their negative counterparts. Detection through polymerase chain reaction (PCR) has been considered as the "gold standard" but imposes burden in low resource settings. Therefore, in the present study, we assessed the validity of cytomorphological features for the detection of HPV in oral leukoplakia (OL), oral squamous cell carcinoma (OSCC), and oropharyngeal squamous cell carcinoma (OPSCC). This study included 63 subjects comprising of 25 OL, 26 OSCC, and 12 OPSCC cases. Exfoliated cells were collected and processed for PCR followed by Papanicolaou staining and subsequent grading. Additionally the non-classical signs were evaluated and statistical analysis included Chi-square and Spearman's test. 23/63 (36.5%) cases showed PCR positivity for HPV16. Most of the cytomorphological features showed significant correlation for the presence of HPV. A greater sensitivity and specificity was observed in the Bethesda system for reporting cervical cytology (TBS) than the Papanicolaou grading system. We conclude that the non-classic cytological features could be employed in the detection of HPV in low resource settings with improved sensitivity. Liquid based cytology graded using TBS could be suitable for oral cytology in the detection of early atypical changes.

A comparative study for qualitative and quantitative analysis of light and fluorescence microscopy stains in oral premalignant and malignant lesions.

Objectives:The present study highlights the use and benefit of fluorescence microscopy stains in easy diagnosis of oral premalignant and malignant lesions. This study was carried out to evaluate the efficiency of light microscopy using Papanicolaou stain (PAP) and May Grunwald Giemsa (MGG) stains and fluorescence microscopy using acridine orange (AO) and 4′,6-diamidino-2-phenyl indole (DAPI) stains. All smears have been qualitatively and quantitatively analyzed in premalignant and malignant lesions using cytology and cytomorphometry.Materials and Methods:The subjects included in this study were selected from patients suffering from premalignant and clinically suspicious malignant lesions. Specimens were collected from the most representative site in study groups and normal buccal mucosa in the control group. The smears were evaluated under fluorescence microscopy for AO and DAPI and under light microscopy for conventional PAP and MGG. We studied 5 HPF to compare the number of normal cells to dysplastic cells. These were then subjected for cytomorphometric and statistical analysis.Results:Assessment of quality using quality index revealed that PAP has better diagnostic efficiency as compared to MGG because the quality index of PAP was 0.87 and that of MGG was 0.74, the quality index of AO was 0.9, and that of DAPI was found to be 0.82, hence proving AO to be better in comparison with DAPI.Conclusion:We found that among all the four stains used AO is reliable, has greater efficacy, and can be easily performed with less cost. Despite the difficulties mentioned in this study, the progress in the field of fluorescence microscopy is expected to be rapid in the following years.

BETEL QUID (PAAN) CHEWING AND ORAL EPITHELIAL DYSPLASIA IN LAHORE: AN ANALYTICAL STUDY

<h3>Background</h3> Betel quid (locally–called "paan") is widely chewed habitually in Pakistan and South and Southeast Asia. It is hand-rolled at paan stalls by smearing a betel leaf (from Piper betel) with slaked lime, catech paste, chopped areca nut, and a variety of condiments and flavoring agents. Paan is considered a group 1 carcinogen for humans by the International Association for Research on Cancer. It is one of the major causes of oral cancer in these regions. The reactive oxygen species released during chewing can initiate carcinogenesis by inducing mutagenesis. <h3>Objective</h3> The present study was performed to determine the frequency of epithelial dysplasia among habitual paan chewers. It also calculated the association between the quantity and quality of paan chewing and the severity of oral epithelial dysplasia. <h3>Methods</h3> This was a cross-sectional analytical study. Cytological smears from 300 habitual paan chewers were collected. After fixation with alcohol, smears were then air dried and stained with hematoxylin, eosin, Giemsa, and Pap stains. The material was then examined under an optical microscope. Epithelial dysplasia was identified and graded using the method of Speight. <h3>Results</h3> Epithelial dysplasia was observed in 57.7% of the habitual paan chewers, which was significantly associated with the number of paans consumed per day as well as duration of paan consumption. These variables were positively correlated with grading of epithelial dysplasia. <h3>Conclusions</h3> The study concludes that epithelial dysplasia was present in the majority of habitual paan chewers in Lahore city. The correlation between the amount of betel quid chewing and the severity of epithelial dysplasia means the effect is dose-dependent.

Role of exfoliative cytology in diagnosis of oral lesions: With special reference to rule out malignancy

Oral malignancy is very common in India because of the excessive use of tobacco chewing and smoking. Procedure of Cytology is inexpensive and unchallenging that can be carried out effortlessly at outdoor patient department to diagnose malignancy at early stage. The present study was carried out to detect cancer pre-invasive stage by use of exfoliative cytology and to probe the probability of using this technique in diagnosis of other oral lesions thought as premalignant ones. A total 102 patients referred from OPD and Surgery Department of a Private Hospital, Dispensary and Cancer Hospital, Delhi, between August 2019 to April 2020 , were included in this study. Two smears were taken by scrap method, from each patient and were prepared after air dried stained with RAPID PAP stain. According to the Papanicolaou classification the smear were assessed along with clinical, epidemiological data and classified in group I to IV. In our result 25 cases were normal, leukoplakia 47 cases with mean age 47.5 years, 85% male. Predominantly 53% Smear show anucleated squames. 06 cases were Oral submucous fibrosis ,66.7% were female with mean age 38.3 years and, the present smear revealed rarification of nuclei in 66% cases. 02 cases of mucosal hyperemia (Erythema), one-one case of traumatic ulcer and granular buccal mucosa were included. Out of 12 cases of malignancy, male were 75% with mean age of 46 year. Buccal mucosa and tongue were the common site with incidence of 60%. The smear showed inflammatory cells in 100%, malignant cells in 75% cases either in groups or in singles. The third type of cells 60%, 37% the undifferentiated cells, and 5% cases Tadpole cells. In this study 75% cases were positive for cancer, 10% were suspicious for cancer and remaining 15% were given as negative for cancer. For the diagnosis of presence or absence of malignancy in a lesion with high accuracy rate Cytology is reliable diagnostic tool. The oral cytologic technique is effortless to do and can provide the help of surgeon/physician, instead of performing an invasive procedure, like a biopsy, or desire more information regarding a lesion before referring the patient.

The Possibility of Analyzing Endometrial Receptivity Using Cells from Embryo Transfer Catheters.

It is very important to investigate the expression of endometrial receptive markers in the endometrium during implantation. Therefore, we examined whether it would be possible to analyze endometrial receptivity using cells from embryo transfer catheters. A total of 81 cycles from 81 consenting patients were enrolled in this study. The tip of the embryo transfer (ET) catheter was cut and immersed in a dedicated reagent. Confirmation of cell distribution was carried out using a Papanicolaou stain and immunocytochemistry. Protein expression was carried out by immunocytochemistry. The expressions of estrogen receptor α, progesterone receptor, and homeobox A10 mRNA were analyzed using quantitative reverse transcription-polymerase chain reaction. We analyzed the relationship between the gene expression profiles associated with pregnancy from endometrial cells. Samples collected from the ET catheter showed clear staining for endometrial cells. Most of the cells were endometrial epithelial cells. Cervical cells were not observed. The protein expression was also confirmed. Three genes were analyzed that are associated with endometrial receptivity. Progesterone receptor expression was 1.4-fold (p<0.05) and homeobox A10 was 2.8-fold (p<0.01) higher in patients who became non-pregnant group, compared to the pregnant group. Estrogen receptor α expression tended to be higher in the non-pregnant group (p=0.18). Our results suggest that endometrial receptivity can be evaluated using cells obtained from the ET catheter. This method may be useful for elucidating the cause of implantation failure by comparing a receptive and non-receptive endometrium at the time of ET.

Evaluation of Micronuclei and Cytomorphometric Changes in Patients with Different Tobacco Related Habits Using Exfoliated Buccal Cells.

Background:Tobacco is one of the main reasons behind the occurrence of oral cancer. Oral cancer, even though being the tenth most common cancer in the world, gets diagnosed at an advanced stage and ends up with poor prognosis. So early diagnosis is the need of the hour. Our study aimed to evaluate the genotoxic changes in patients with different tobacco habits using buccal exfoliated cells. Methods:Buccal smears were taken from smokers (30), smokeless tobacco users (30), combined tobacco users (30) and controls (30) with clinically normal oral mucosa. All the smears were stained with Papanicolaou stain and Feulgen stain and viewed under light microscope for the evaluation of mean number of micronuclei, mean micronuclei per cell, frequency of cells showing micronuclei, nuclear area, cytoplasmic area, nuclear-cytoplasmic ratio. Results:Mean number of micronuclei, mean micronuclei per cell, frequency of cells showing micronuclei, and nuclear area were significantly increased in tobacco users than controls, especially in combined tobacco users. Nuclear-cytoplasmic ratio was increased and cytoplasmic area was decreased in tobacco users than controls. Conclusion:Tobacco in any consumable form is genotoxic. Smoking and smokeless tobacco, when consumed together, synergistically causes higher genetic damage. Different tobacco habits have different deleterious effects on oral mucosa, and these effects are more pronounced when the patients have combined habits. So, detecting the genotoxic changes through exfoliative cytology can be used as a simple yet reliable marker for early detection of carcinogenesis.

A Retrospective study on the advantage of Fine Needle Aspiration Cytology (FNAC) as Walk- in Diagnostic Test in Patients with Cervical Lymphadenopathy

Objective: To determine the etiologic spectrum of cervical lymphadenopathy using fine-needle aspiration cytology (FNAC) in a tertiary care setting. Methods: In the present retrospective study, we presented the data of 1773 patients who presented in Multan Medical and Dental College within 11 years from Jan-2007 to Dec-2018. In all patients, FNAC was performed using a 22-23-gauge needle by using a 10 ml syringe. After collection specimens were sent to the hospital laboratory, the collected samples were stained using Papanicolaou stain, the Field’sbstain, and ZiehlaNeelsen (ZN) stain and acid-fast bacilli (AFB) for determination of the pattern of cervical lymphadenopathy. Specimen analysis was done in a private diagnostic center. Results: Mean age of study participants was 37.8±9.7 years. There were 1012 (57.07%) male patients and 761 (42.93%) female patients. Tuberculosis was diagnosed in 655 (36.9%) patients, metastasis in 349 (19.6%), reactive hyperplasia in 267 (15.0%), acute lymphadenitis in 193 (10.9%), lympho-proliferative lesions in 96 (5.4%), non-hodgkin lymphoma in 30 (1.69%), chronic infections in 18 (1.0%) and Hodgkin lymphoma in 11 (0.62%) patients. Conclusion: In the present study, cervical tuberculosis was the most common diagnosis found in 36.9% of cases, which present that tuberculosis is very common in our population. FNAC is easy and can be performed as a bedside procedure for the diagnosis of cervical lymphadenopathy. Keywords: Cervical lymphadenopathy, Fine needle aspiration cytology.

To study cytomorphological details in case of autoimmune thyroiditis

Various smears were readied, air-dried and fixed in ethanol for staining by May-Grunwald-Giemsa stain and Papanicolaou staining strategy. A nitty gritty assessment of the cytologic smears was done and includes like cellularity, sum and nature of colloid, Hurthle cell change, anisonucleosis of follicular cells, range of receptive lymphoid cells and other incendiary cells like eosinophils, macrophages, goliath cells and epithelioid cells were noted. Thyroid capacity tests were finished utilizing COBAS E analyser.In our study we found, 82% Females whereas 18% Males. Lymphocytic Thyroiditis 42%, Granulomatous Thyroiditis 31 &amp; Hashimoto’s Thyroiditis 27%. Types of swelling and diagnosis with painless 71 % and painful 29 %. Cross-sectional Observational Study. A total of 200 thyroiditis cases studied with detailed history, thorough clinical examination and relevant blood investigations like Thyroid Profile were being done. The various studies on the correlation of cytomorphological spectrum and antithyroglobulin antibodies in autoimmune thyroiditis cases all acreoss the globe have great variations in their findings.

Novel Biallelic DNAH1 Variations Cause Multiple Morphological Abnormalities of the Sperm Flagella.

Sperm motility is vital to human reproduction, and malformed sperm flagella can cause male infertility. Individuals with multiple morphological abnormalities of the flagella mostly have absent, short, coiled, bent, and/or irregular-caliber flagella. In this study, a patient with male infertility underwent a physical examination along with his wife. Genetic testing was performed by whole-exome sequencing of the couple, and Sanger sequencing was performed for validation. Novel biallelic variations in the DNAH1: (NM_015512.4) gene consisting of c.1336G>C (p.E446Q) and c.2912G>A (p.R971H) were identified. In silico structural analysis revealed that the amino acid residues affected by the variation were evolutionarily conserved, and the variant p.R971H influenced the stability of the DNAH1 protein. Morphological studies of the patient's sperm showed defects in its flagella. Results of Papanicolaou staining and scanning electron microscopy demonstrated coiled and short flagella with multiple anomalies. Transmission electron microscopy of the sperm flagella showed that the inner dynein arm and radial spoke were absent, and the dense fiber and microtubule doublets were displaced. Quantitative PCR of the mRNA of the patient's sperm showed that the expression of DNALI1 was dramatically reduced. Collectively, these findings elucidated the genetic cause of the family's infertility and provided insight into the functioning of the DNAH1 gene.

English

BACKGROUND Semen analysis is an integral part of work up for infertility in men, with sperm morphology being an important qualitative parameter. Qualitative defects can affect any part of the sperm and are classified as defects in the head, middle piece, and tail, based on morphology. The focus of the study was to assess qualitative defects in sperms by light microscopy, in semen with normal sperm counts. METHODS This study is hospital based, descriptive, retrospective study. Of the semen samples received in the clinical laboratory, fifty with normal sperm counts were included in the study and processed according to standard protocol. For evaluation of qualitative defects by sperm morphology, smears were fixed in ethanol, stained with Papanicolaou stain [PAP], and assessed under light microscope. RESULTS The 50 semen samples included in the study had sperm counts ranging from 15 to 80 million / ml. Thirty samples had less than 10 % abnormal forms, fourteen samples had 11 - 20 % abnormal forms, five samples had 21 - 30 % abnormal forms and one sample had 40 % abnormal sperms. Qualitative defects were classified as morphological abnormalities in head, neck, and tail. Of the fifty cases, most defects were found in the head, followed by those in the neck and tail. Common defects noted were double head (44 %), abnormal sized heads, and bent neck (48 %). Coiling was a common defect noted in the tail (10 %). Most sperms showed a combination of defects. CONCLUSIONS Qualitative defects in sperm morphology are often seen in samples with normal sperm counts. Assessment of microscopic characteristics of human spermatozoa is as important as count and motility in the complete evaluation and work-up of semen samples in cases of infertility. KEY WORDS Semen, Sperm, Quality, Microscopy, Morphology