Papanicolaou Staining Technique Research Articles

Comparative Study of Modified Ultrafast Papanicolaou and Standard Papanicolaou Staining Technique for the Assessment of Cervical Smears

Introduction: Cervical invasive carcinoma is one of the most frequent cancers in the world, and it can be prevented with screening. The papanicolaou (Pap) smear is the most common screening procedure. Pap staining, as it is now done, is costly, requires a large amount of alcohol, and takes a long time. Materials and Methods: A total of 170 patients were recruited. One hundred and sixty-six patients, considering eligibility criteria, undergoing Pap smear test. Eligibility criteria: Age: 18–49 years females, who did not have a total hysterectomy done, who consented to participate in the study. Each patient had two smears taken on glass slides. Standard Pap smears were fixed for at least 15 min in 95% ethanol. The other smear was air-dried before being fixed with alcohol formalin and stained with modified ultrafast Pap (MUFP) stain. A comparison of two different techniques was made on the basis of cytomorphological features. Results: In studied smears, a clean background was seen in 18.24% and 14.12% of the cases in Pap stain and MUFP, respectively. Kappa analysis found a good correlation between Pap stain and MUFP (kappa: 0.81, P < 0.01). Crisp chromatin was seen in 89.41% and 67.65% of MUFP and Pap staining, respectively. About 88.82% of MUFP stains showed optimal cytoplasmic details and the same was revealed by 81.18% of Pap stains. MUFP has a better quality index (QI) as compared to Pap staining as no case in MUFP had QI of < 0.80 with a statistically significant difference as P < 0.05. Conclusion: MUFP is a simple, user-friendly, affordable, and less time-consuming alternative in low-resource areas in comparison to the traditional technique for mass cervical cancer screening.

Oncoguía de cáncer cervicouterino 2023

Guidelines on cervical cancer (CC) enable a series of recommendations and strategies for primary and secondary prevention, early diagnosis, in advanced stages, as well as treatment in various scenarios. The human papillomavirus (HPV) vaccine can reduce the development of cervical precancerous lesions. That will drastically lower the incidence rates of CC. The tool traditionally used for secondary prevention is cytology with the Papanicolaou staining technique. The CC treatment will be determined based on the initial stage: for early stages, it is based on surgical management; in locally advanced disease, treatment requires concomitant chemo - radiotherapy + brachytherapy (BT). Recent modifications in staging, standardization in surgical management, and various options for recurrence or metastatic disease have been added.

Detection and Comparision of Cytopathological Changes in Smokers and Non-Smokers: A Comparative Study

Background: Oral cancer is one of the sixth most common cancers in the world. Oral exfoliative cytology is one of the popular screening tool for oral cancer. Use of tobacco in any form are documented as the most common cause as initiators for dysplastic changes in oral mucosa. The purpose of the study was to detect the cytological changes in buccal mucosa, tongue and palate among non-smokers smokers. Material Methods: Smears sample were collected according to site (buccal mucosa, tongue palate) from 100 subjects among smokers non-smokers. Smears were then stained using Papanicolaou staining technique. Results: Among the smokers and non-smokers the results were statistically significant. Conclusion: Recent advances in the clinical visualization and detection of the oral mucosa have made the viability of cytological procedures more specific and sensitive. Contact endoscopy and use of autofluorescence devices are the forerunners in this group. The fluorescence characteristics of tissues depend upon their biochemical composition and histomorphological architecture, both of which undergo a change during malignant transformation. These changes are detectable as an alteration in the fluorescence spectral profile of the tissues21. Due to low feasibilities of such devices the benchmark of diagnosis will be microscopic tissue examination. Hence cytological smears will always be highly specific, sensitive, easy to use and reproducible procedures in routine screening of population for potentially and malignant conditions of the oral cavity.

Cervicovaginal detection of microorganisms in patients with type 2 diabetes

The purpose of the present investigation was to report the presence of cervicovaginal microorganisms most frequently found in cervicovaginal exfoliative smears (Papanicolaou) of patients with type 2 diabetes. Cervical-vaginal exfoliative cytologist was performed on an open population to all patients who attended from March 2010 to March 2020 in the within the framework of the early cancer detection program, gynecological interrogations were applied to 1587 patients who, after taking cell samples, were processed using the Papanicolaou staining technique modified by the same for subsequent microscopic diagnosis. Of the results obtained from the 1587 patients studied, 148 (9.32%) suffer from type 2 diabetes mellitus and of these only 130 (87.83%) showed at least one microorganism under microscopic observation, being Candida spp., the most found microorganism in samples from patients with type 2 diabetes. In conclusions, type 2 diabetes is a disease that predisposes to the appearance of microorganisms and cervicovaginal infections, particularly by Candida spp., therefore, there is a need for surveillance guidelines and efforts to reverse this disease and its complications.

A study of cytopathological pattern of cervical papanicolaou smear examination in women between 20 - 75 years of age group

Background: Cancer of uterine cervix is a leading cause of mortality and morbidity among women worldwide. The primary cause of this cancer is infection with human papillomavirus. Pap smear is a simple, safe, non-invasive and cost effective method for the detection of pre-cancerous, cancerous and benign lesions of cervix.Aim: To study the cytological patterns of cervical Pap smears in 766 cases and to classify the cytological abnormalities based on the Bethesda system 2014.Materials and methods: A retrospective study of total 766 cases of cervical smears was done at Pathology department, GMERS Medical College and Civil Hospital, Sola, Ahmedabad over a period of 1.5 years starting from 1st January 2020 to 30th June 2021. Women between 20 -75 years of age group were included in the study. Cervical smears were taken by obstetrics and gynaecology department and were received at pathology department, stained by using rapid Papanicolaou staining technique and studied by using light microscopy.Results: we studied total 766 cases out of which 26(3.40%) smears were unsatisfactory for evaluation. Out of these 740 cases, most common were inflammatory lesions-382 cases (49.86%).Cervical epithelial cell abnormalities including ASCUS-5 cases (0.67%), ASC-H -2 cases (0.24%), LSIL- 6 cases (0.78%), HSIL-1 case (0.13%) and AGC-NOS – 1 case (0.13%) were detected in our study. Majority of the cases were belonging to 31-40 years of age group -364 cases(47.50%) followed by 41-50 years-260 cases (33.94%).Conclusion: Pap smear is an easy, simple, safe and noninvasive method of cervical examination. By using Pap smear as a screening test, it is possible to detect malignant lesions at an earlier stage.

Tobacco Consumption in Some Traveling Agencies in Bafoussam in West Region Of Cameroon, Associated To Mouth Cytology and Malondialdehyde (MDA) Profile

Tobacco is mostly regarded as one of the primary etiologic factor in causing oral cancer. Its use affects the pattern of maturation in oral mucosal cells and results in changes in the appearance of the tissues. Lipid peroxidation (LPO) has been implicated in the pathogenesis of several pathologic disorders, including oral premalignant and malignant lesions. The aim of this study was to determine the prevalence of tobacco use and assess its effects on oral epithelial cells and oxidant/antioxidant balance in the oral cavities of tobacco consumers in some travelling agencies in Bafoussam West Region Cameroon.
 A cross sectional study with a total of 120 individuals (divided into two groups; group A (n=55) including non-consumers of tobacco above 21 years old and group B (n=65) including tobacco consumers above 21 years old), voluntarily enrolled in the study. After giving their consents and answering a series of questions from questionnaires, saliva samples and buccal smears were collected from their oral cavities. Buccal smears were stained using Papanicolaou staining technique and the presence of hyperkeratosis, tobacco hyperkeratosis, non-specific inflammatory changes and oral microflora were assessed. [MDA] was determined using the Thiobarbituric acid method. The results showed that male tobacco consumers made 48.3% and most of them were aged between 21-31 years old, while female tobacco consumers made 5.7% of the total sample population. Smoking was the method most used by consumers (41.7%) and 66% of them smoked for more than 10 years while 50% smoked more than 10 sticks of cigarette per day. A statistical significant relationship was found between tobacco consumption and oral epithelial changes (p<0.001). Only tobacco consumers presented with tobacco hyperkeratotic buccal smears (20%) when compared with the control group. A good number of hyperkeratotic buccal smears (39.2%) was found in both the case (17.5) and control (21.7%) groups. 24.1% of the total sample population had normal buccal smears, where 8.3% were consumers and 15.8% were non-consumers. Microorganisms were found on buccal smears of 5.9% of individuals and 9.2% had non-specific inflammatory changes. Between tobacco consumption and salivary [MDA] in saliva of tobacco consumers was significantly higher compared to non-consumers (p<0.001). A strongly positive correlation was found between salivary [MDA] and oral epithelial changes (r=0.204, p=0.026 at 0.05CI). In conclusion, tobacco consumption severely increases the risk for developing oral mucosal proliferative lesions, and the level of oxidative stress in the saliva of individuals in the study population.

Evidence of genotoxicity and cytotoxicity of X-rays in the oral mucosa epithelium of adults subjected to cone beam computed tomography.

Evidence of genotoxicity and cytotoxicity of X-rays in the oral mucosa epithelium of adults subjected to cone beam computed tomography.

Short-duration Papanicolaou stain (SPS)—an alternative to conventional Papanicolaou stain in routine cytopathology?

Papanicolaou (PAP) stain is the most common stain used for cytological evaluation. Judicious use of the PAP stain results in a well-stained nuclear chromatin, cytoplasmic transparency, and differential cytoplasmic staining which helps the pathologists in the diagnosis of various lesions. The major disadvantage of the conventional PAP stain is the long duration of the staining process. Since cervical cytology is used as a screening method for diagnosis of cervical cancer, it becomes essential to device a modified stain which gives rapid results. Hence, this study was undertaken to study the utility of a rapid-modified short-duration PAP stain. This is a prospective study conducted on 100 patient samples which were received in the Cytopathology Laboratory of Sree Balaji Medical College and Hospital, Chromepet, Chennai, India. Two PAP smears were collected from the study population and one was stained with the conventional PAP staining technique and the other was stained with the modified rapid-short PAP staining method. The staining characteristics of these smears were evaluated and compared. Smears stained by both the staining techniques were evaluated using a scoring system which assesses the nuclear, cytoplasmic, and the background staining characteristics. It was found that the rapid-short PAP staining technique is found to be more effective compared to the conventional PAP stain with a P value of < 0.0001. Short-duration Papanicolaou staining technique could be a definite better alternative to the conventional PAP stain both by being cost effective and by consuming less time for staining.

Modified ultrafast Papanicolaou staining technique: A comparative study.

Introduction:Ultrafast Papanicolaou stain (UFP) was introduced as a hybrid of Romanowsky and Papanicolaou (PAP) stain. It enhances the quality and reduces the time. In the present study, a modified staining technique was adapted where Gill's Hematoxylin was replaced by Harris Hematoxylin.Aims:The aim of the study was to assess the use of the modified ultrafast Papanicolaou (MUFP) stain for fine needle aspiration cytology (FNAC) of head and neck swellings in comparison with the routine PAP stain, hematoxylin and eosin (H and E), and Giemsa.Materials and Methods:Forty FNACs of head and neck swellings were collected. FNAC procedure was performed by standard method; two smears were fixed in 95% propanol and stained with PAP and H and E. Two smears were air dried, 1 was stained with Giemsa, and 1 was rehydrated with normal saline, fixed in alcoholic formalin, and stained with MUFP. Four parameters were considered and scored background, cell morphology, nuclear staining, and overall staining pattern).Results:The quality of MUFP smears were better when compared to routine PAP, H and E, and Giemsa, and was statistically significant by Wilcoxon matched pair test.Conclusions:MUFP stain in comparison to routine PAP, H and E, and Giemsa provides an excellent and suitable alterative in cytological staining for the study of various organs.

Molecular detection of human papillomavirus from abnormal cervical cytology of women attending a tertiary health facility in Ido-ekiti, southwest Nigeria.

Human papillomavirus (HPV) has been implicated as one of the principal causes of cervical cancer, which is the second highest cause of cancer deaths among Nigerian women. This study was aimed at determining the presence of HPV DNA in abnormal cervical cytology of a group of women who were screened using Papanicolaou staining technique. A total of 200 women attending the Obstetrics and Gynaecology clinic of the Federal Teaching Hospital, Ido-Ekiti, were screened by means of conventional Pap smear screening, while positive samples underwent molecular analyses by means of DNA isolation techniques and polymerase chain reaction (PCR). Results revealed that 14 (7%) of the subjects were positive for abnormal cytology. Abnormalities found among the subjects included: low-grade squamous intraepithelial lesions (LSIL), which constituted 50% of the total abnormal smears, high-grade squamous intraepithelial lesion (HSIL) and atypical squamous cells of undetermined significance (ASCUS), which were 28.6% and 21.4%, respectively. Molecular analyses showed that all the samples from abnormal cervical cytology subjected to HPV DNA extraction and gene amplification contained HPV DNA. The high prevalence of HPV DNA in abnormal cytology gives credence to the fact that the presence of HPV is a critical indicator of the development of cervical cancer. Thus more effort should be put into vaccine production and distribution in order to reduce the incidence of cervical cancer in Nigeria.

Fine Needle Aspirate and Cytology (FNAC) as Useful Tool in the Diagnosis of Suspected Tuberculous Lymphadenitis in Rwanda, 2009

Background: The number of people infected with tuberculosis is gradually increasing in Rwanda. The diagnosis of suspected Tuberculous Lymphadenitis (TL) is still difficult to diagnose and remain expensive when applying surgical procedures. Objective: To study the usefulness of fine needle aspirates in diagnosis of TL in patients from a low income country. Methods: Triplicate smears from lymph node aspirates were prepared. Air-dried smears were stained by hot ZN staining technique for AFB examination, and Gram staining technique for exclusion of any bacterial infection and Papanicolaou staining technique was done for cytological to detect malignant cells and other pathology. Slides were examined by laboratory technologist and pathologist respectively. Results: A total number of 138 specimens from suspected TL patients were analysed, of which 14 (10.1%) were ZN positive while cytology revealed 25 (18.1%) cases of Tuberculous Lymphadenitis. From Papanicolaou stain 25 (18.1%) cases were either supportive for TB (7.2%) cases or suspicious for TB (10.9%) while 113 (81.9%) there was no features for TB. Among 25 cases which were supportive or suspicious for TB only 6 cases (4.3%) were also ZN positive and 19 (13.8%) were ZN negative. Cytology revealed also 19 (13.8%) cases of reactive nodes, 25 (18.1%) accounted for other pathologies and 33 (23.9%) were inadequate samples for analysis and interpretation. Gram stain showed 2 (2.7%) cases of Gram positive cocci. Conclusion: In low income countries, the use of FNA cytology with more training should be considered as a useful tool in diagnosis of TL instead of biopsy applying surgical procedures.

Comparison of the Routine Papanicolaou Staining Technique with the Rapid, Economic, Acetic Acid, Papanicolaou (REAP) Technique

Background : The universal stain for oral cytological screening for precancer and cancer of oral cavity is Papanicolaou stain which has been used in different laboratories with many modifications. Objectives : To assess the superiority of staining of smears by REAP technique compared to PAP technique. Material and Methods: 100 smears were collected from 50 patients. One set of smears were stained with conventional PAP technique and the other set with the REAP technique. In the REAP technique, the ethanol bath in the pre Orange G6, post Orange G6 and post EA50 stages is replaced by 1% acetic acid; Tap water is used instead of Scott's tap water and hematoxylin is pre heated in water bath to 60°C for rapid penetration. Methanol is used for final dehydration. The two sets of smears are observed by two independent observers and assessed for the optimal and suboptimal nuclear and cytoplasmic staining. The results are compared and analyzed statistically. Results : Good cytoplasmic transparency and optimal nuclear details were seen in REAP stained smears compared to the conventional PAP smears. The cost involved in REAP method was lesser compared to conventional PAP technique. REAP technique took 3 minutes for completion compared to PAP technique that involves a minimum of 20 minutes. Conclusion: REAP technique produces better stained smears that are cost effective and involve minimal time for mass screening of oral cancer as compared to conventional PAP smears.

Correlation Between &lt;i&gt;Koilocytes&lt;/i&gt;, HPV DNA and CD4 Cells Count in Male and Female HIV Cohorts

The study x-rays the relationship between <i>K</i><i>oilocytes</i> and [Cluster of Differentiation]CD4 Count from HIV positive unisex cohorts aged 15 -40 years and above. The cohorts, were randomly tested for the presence of koilocytes, using their semen and endocervical swab samples respectively at some top Hospitals in Abuja, F.C.T. Nigeria. Their blood samples were tested for CD4 count. Papanicolaou staining technique was adopted-for demonstrating koilocytes. Results showed that, high koilocytes count of 39.0% and low CD4 Count of 350 lymphocyte counts /µl were observed; and low monthly mean koilocytes count of 7.60% and high CD4 Count of 687 lymphocytes counts/ µl were observed in the female and male cohorts respectively. High monthly prevalence count of koilocytes and low CD4 Count in the female cohorts and verse versa in the male cohorts were significantly correlated (P<0.05). Depression of immunity, is worsen in HPV and HIV co- infectivity.

Papanicolaou Stain

To standardize an inexpensive and rapid Papanicolaou staining technique with limited ethanol usage.Smears from 200 patients were collected (2 per patient) and fixed in methanol. Half were subjected to conventional Papanicolaou and half to stain ing with rapid, economical, acetic acid Papanicolaou (REAP) stain. In REAP, pre-OG6 and post-OG6 and post-EA36 ethanol baths were replaced by 1% acetic acid and Scott's tap water with tap water. Hematoxylin was preheated to 60 degrees C. Final dehydration was with methanol. REAP smears were compared with Papanicolaou smears for optimal cytoplasmic and nuclear staining, stain preservation, cost and turnaround time.With the REAP method, cytoplasmic and nuclear staining was optimal in 181 and 192 cases, respectively. The staining time was considerably reduced, to 3 minutes, and the cost per smear was reduced to one fourth. The staining quality remained good in all the smears for > 2 years.REAP is a rapid, cost-effective alternative to Papanicolaou stain. Though low stain penetration in large cell clusters is a limitation, final interpretation was not compromised.

Nonsperm cells in human semen: with special reference to seminal leukocytes and their possible influence on fertility.

Nonspermatozoal cells (NSC) in semen, including immature germ cells, leukocytes (WBC) and epithelial cells, are normally found in a concentration less than 15% of the sperm concentration. Cytological staining methods and immunoenzymatic assays for characterization of seminal WBC and methods for detection of various enzymes from WBC are evaluated. Criteria for abnormally high levels of WBC in semen, caused by exogenic factors (e.g., infection) and possibly by endogenic factors (e.g., abnormal sperm), are discussed. Leukocytospermia is associated with reduced in vitro fertilization rate, and experimentally-measured sperm functions (e.g., motility) are inhibited by high concentrations of certain WBC products (e.g., reactive oxygen species and interferon-gamma). At the present stage, the following procedures could be used to evaluate WBC in ejaculates from infertile patients: (1) count the number of NSC using a hemocytometer. (2) Determine the number of WBC in percentage of NSC in smears using the Papanicolaou staining technique, or an immuno-enzymatic assay using monoclonal antibodies to the different kinds of WBC. (3) If, abnormally high levels of WBC (according to the WHO criteria or based on a percentile determined from examination of normal, fertile men) is detected, a second ejaculate should be examined 3 months later. (4) Persistent leukocytospermia should indicate physical and microbiological examinations, after which antibiotic treatment may be indicated.

Flow Cytometric Analysis of DNA Content in Cervical Lesions

Flow cytometry is used to screen gynaecologic specimens for Cervical Intraepithelial Neoplasia (CIN) and its precancerous lesions, possibly associated with human papilloma virus (HPV), in order to investigate the role of aneuploidy as a biological marker in HPV and CIN lesions. A total of 299 cervical samples was collected by scraping and the cellular DNA content was measured using the propidium iodide staining procedure. Two groups of patients were studied; a group of 142 negative controls for cytology and groups of patients assigned to mild, moderate or severe dysplasia, carcinoma in situ diagnosed by histologic classification according to the Papanicolaou staining technique. Pathological samples showing an alteration of the DNA index or perturbation of the cellular phase S compartment ranged from 6.4% to 41.9%. Our results confirm those obtained by other authors and suggest the hypothesis that aneuploidy is present with permanence of viral DNA in early stages of carcinogenesis, which can be used as a marker in the transition from benign to malignant cells. This work is of potential interest for the possible follow-up of patients having condyloma and could aid the early diagnosis of cervical carcinoma.

Vitrectomy. Clinical data and cytologic findings of vitrectomy specimens.

Fifty vitreous specimens obtained by routine pars plana vitrectomies were examined using Millipore filter processing and Papanicolaou staining technique. Cytological findings comprised of macrophages (melanophages, siderophages) and inflammatory cells, mainly lymphocytes. In a few cases, pigmented an nonpigmented epithelial cells of the ciliary body were encountered, retinal sensory cells only in one case. Fibroblasts and fragments of granulation tissue occurred in four of the 14 diabetic cases, in four of the 21 posttraumatic cases, in one case of chronic uveitis as well as in one case of retrolental fibroplasia. Fragments of ciliary body were in three specimens from injured eyes. In one case of chronic uveitis slight suspicion of ciliary body melanoma was aroused on the basis of distinct cellular atypia. There was agreement between clinical and cytological diagnosis in most cases, but particularly in posttraumatic and inflammatory diseases cytologic examination offered new diagnostic data. This is a new method of microscopical verification of the condition in the vitreous body in various diseases. Our experiences are preliminary but we feel that this method has proven its availability.

Low-speed purification of human placental nuclei

A simple method for the purification of human placental nuclei is described. Nuclei were isolated by homogenizing tissue in standard saline citrate solution in the presence of zinc chloride to stabilize the nuclear membranes, NP40 as non-ionic detergent and sodium bisulphite for inhibition of proteolytic activity. Nuclei purification was achieved by low-speed centrifugation through a discontinuous sucrose gradient. The purified nuclei were evaluated by morphological criteria using phase contrast and electron microscopy. The extent of contamination by cytoplasmic debris was estimated by Papanicolaou's staining technique. Biochemical criteria include measurements of alkaline phosphatase activity as a plasma membrane enzyme marker and DNA-dependent RNA polymerase activity for the functional integrity of nuclear components. Transcriptionally active nuclei were obtained but the yield of nuclei was low; however, this low yield is compensated by the high degree of purity, the simplicity of the method and the functional and morphological integrity of the purified nuclei.

Papanicolaou staining technique for plant cells.

(1973). Papanicolaou Staining Technique for Plant Cells. Stain Technology: Vol. 48, No. 5, pp. 259-260.

Specific Epithelial Degeneration (Ciliocytophthoria) in Inflammatory and Malignant Respiratory Disease

SUMMARY 1Ciliocvtophthoria is a specific form of degeneration of columnar ciliated epithelium of the respiratory tract. 2Using the Papanicolaou staining technique for the examination of sputum, CCP cells are easily recognized by the formation of tufts, pyknotic and clumped nuclei, and by the presence of acidophilic inclusion bodies in the cytoplasm and nucleus. 3A study of 301 cases with varied respiratory diseases showed the highest incidence of CCP to occur.in patients with acute viral infections and bronchogenic carcinoma. 4Among 77 cases positive for CCP, 65 (84 per cent) occurred in patients with either viral or malignant respiratory disease. 5The relationship of CCP formation to viral disease and malignancy is explained by an epitheliotropic process occurring in both conditions resulting in the desquamation, and subsequent degeneration, of large numbers of columnar ciliated epithelial cells.