Modified Papanicolaou Stain Research Articles

Micrometastasis detection using modified papanicolaou stain in nodal tissues of oral squamous cell carcinoma - A histological study.

Cervical lymph node metastasis is the most important prognostic factor in Squamous Cell Carcinoma of Head and Neck (SCCHN). Detection and evaluation of micro-metastasis forms the basis for diagnosis, staging, treatment options and prognosis. Lymph node prognostic factors are extremely important for the survival and recurrence in the patient. Assessing lymph node metastasis in the absence of clinical enlargement is challenging. To evaluate micrometastasis and individual tumor cells (ITC) in regional lymph nodes of oral squamous cell carcinoma (OSCC) by modified papanicolaou (PAP) stain and re-evaluate the tumor staging. The retrospective study was executed at MS Ramaiah University of Applied Sciences. The current study constituted a total of 40 lymph nodes from OSCC patients, metastatic (n=20) and non-metastatic lymph nodes (n=20). All sections were stained with H & E followed by modified PAP stain. Modified PAP was used for identification of micrometastasis deposits. The Chi square test was employed to analyze significance. Modified PAP stain proved to be more accurate (p = 0.006) than H and E stain in detecting micrometastasis which accounted for 15% of non-metastatic lymph node sections used in our study. Special stain like modified PAP stain is valuable and sensitive in detecting micro-metastasis over H and E stain. Detection of micrometastasis in OSCC patients is advantageous for the patient as it influences staging, it modifies the treatment plan in terms of both radiotherapy and chemotherapy.

Efficacy of Visual inspection with acetic acid (VIA) and PAP smear as a screening method for diagnosis of premalignant lesions of cervix in high-risk patient

Background: Carcinoma Cervix is one of the common malignant pathology causing considerable morbidity and mortality in women. One of the important aspects of management of carcinoma cervix is its early detection and appropriate management since delay in the diagnosis can have catastrophic consequences. Unfortunately in developing countries in many cases, diagnosis is delayed due to non-availability of facilities at rural areas. To overcome this screening methods such as Visual Inspection with Acetic Acid (VIA) and PAP smear can be done in high-risk patients and in selected cases cervical biopsy can be done. Aims and Objectives: (1) To compare VIA and PAP smear in relation to colposcopy as a screening method for premalignant lesions of cervix. (2) To study the efficacy of VIA as a primary screening test for detection of premalignant lesions of cervical cancer. Materials and Methods: This was a prospective analytical study in which 100 women with unhealthy cervix attending obstetrics and gynecology out patient department who were willing to participate and gave written informed consent were enrolled in the study on the basis of a predefined inclusion and exclusion criteria. VIA was done. Pap smear was taken and slides were stained with modified Papanicolaou staining method and examined under light microscope. we compared the efficacy of VIA and PAP smear as a screening method for diagnosis of premalignant lesions of cervix in high-risk patient, and the results were confirmed with the help of colposcopy examination and its histopathological correlation with guided biopsy. Results: Mean age of the study population was found to be 40.94 years. When VIA, PAP Smear, and colposcopy were compared highest sensitivity was seen in VIA whereas colposcopy showed highest accuracy (82%). PAP was found to have a higher specificity (84.62%) as compared to VIA or colposcopy. Highest positive predictive value and negative predictive value was seen in PAP smear (78.95%) and colposcopy (86.96%). Conclusion: VIA is an easy to perform and highly sensitive test for diagnosis of carcinoma cervix however it has got a poor specificity. Colposcopy was found to be a better diagnostic test as it was both more sensitive as well as specific to diagnose premalignant lesions.

Homozygous SPAG6 variants can induce nonsyndromic asthenoteratozoospermia with severe MMAF

BackgroundMultiple morphological abnormalities of the sperm flagella (MMAF) is a subtype of severe asthenoteratozoospermia with poorly understood genetic etiology. SPAG6 is a core axonemal component that plays a critical role in the formation of cilia and sperm flagella. Previous studies have reported that mutations in SPAG6 cause primary ciliary dyskinesia (PCD), but the association between SPAG6 gene variants and the MMAF phenotype has not yet been described.MethodsWe performed whole-exome sequencing (WES) in two unrelated Han Chinese men with MMAF. Sanger sequencing was used to validate the candidate variants. Routine semen analysis was carried out according to the WHO guidelines (5th Edition). Sperm morphology was assessed using modified Papanicolaou staining. Scanning and transmission electron microscopy (S/TEM) was performed to observe the ultrastructural defects of the sperm flagella. Western blot analysis and immunofluorescence (IF) of spermatozoa were performed to examine the expression of SPAG6 protein. Assisted fertilization with intracytoplasmic sperm injection (ICSI) was applied.ResultsTwo homozygous SPAG6 variants were identified by WES and Sanger validation in two patients with MMAF phenotype (F1 II-1: c.308C > A, p. A103D; F2 II-1: c. 585delA, p. K196Sfs*6). Semen analysis showed progressive rates of less than 1%, and most of the spermatozoa presented MMAF by Papanicolaou staining. TEM revealed that the overall axonemal ultrastructure was disrupted and primarily presented an abnormal “9 + 0” configuration. No other PCD-related symptoms were found on physical examination and medical consultations, as well as lung CT screening. The level of SPAG6 protein was significantly decreased in the spermatozoa, and IF analysis revealed that SPAG6 staining was extremely weak and discontinuous in the sperm flagella of the two patients. Notably, F1 II-1 and his wife conceived successfully after undergoing ICSI.ConclusionsOur research provides new evidence for a potential correlation between SPAG6 variants and the MMAF phenotype.

Modified Papanicolaou staining for oral swab samples stored long term

ABSTRACT Pathologists collect swab samples for Papanicolaou (Pap) staining to diagnose various diseases including cancer and HIV. Time constraints and limited resources, may preclude staining a sample immediately. To re-confirm results, samples must be frozen for later analysis. We present a method for Pap staining cells that have been stored long term. An effective method for Pap staining of frozen cells should enable flexibility for processing samples.

COMPARATIVE ANALYSIS OF TUBERCULOSIS BACTERIA IN SALIVA AND SPUTUM USING ZIEHL-NEELSEN, PAPANICOLAOU AND BLEACH MODIFIED PAPANICOLAOU INDUCED FLUORESCENT MICROSCOPY

Background: This research contrasted the identification of Mycobacterium tuberculosis in saliva and sputum samples using Ziehl-Neelsen, Papanicolaou, and Bleach Modified Papanicolaou fluorescent microscopy.
 Methodology: Sputum and saliva samples were collected in a clean sterile, leak-proof, wide-mouth containers early in the morning. Each saliva and sputum sample was further divided into 3 groups, Group A: sputum stained by Ziehl-Neelsen staining method, Group B: sputum smears stained by Papanicolaou staining method, Group C: sputum smears stained by bleach modified Papanicolaou staining method. saliva and sputum infected by Papanicolaou and observed under a Fluorescent Microscope for tubercle bacilli..
 Results: The results obtained from saliva stained smears showed 63.3% of cases positive in both BMP and Pap stained saliva samples and 46.6% of cases positive in ZN stained saliva samples. Further grading of the individual staining method was carried out. Scant bacilli; 20% in Pap and 10% in BMP, 1+; 30% in Pap and 33.3% in BMP, 2+;10% in Pap and 6% in BMP and 3+-3.3% in Pap and 13% in BMP. ZN stained saliva samples scant bacilli; 56.6%, 1+; 13%, 2+; 10% and 3+; nil. Sputum samples stained by BMP, Pap, and Zn showed 100% positive results (30/30). But the grading differed among individual staining techniques.
 Conclusion: In our study sputum samples showed 100% positivity in comparison to saliva samples with 63.33%. Though the results of the saliva sample were lower than that of the sputum samples, the BMP method used in our study is first of its kind to the best of our knowledge. It is a simpler, convenient, and economical method that can be used in routine diagnosis of TB.

Correlations of Sperm Mitochondrial Membrane Potential with Semen Parameters and Male Obesity

Background: To investigate the correlations of sperm mitochondrial membrane potential (MMP) with semen parameters and body mass index (BMI) in males with obesity. Methods: Semen samples were obtained by masturbation after 3–7 days of sexual abstinence from males who visited semen collect room of Shanghai Ninth People’s Hospital. Conventional semen analyses were performed by computer-aided sperm analysis (CASA), and sperm morphology was analyzed by modified Papanicolaou staining. Spermatozoa were stained by JC-1 to evaluate MMP through flow cytometry. Results: Sperm MMP of asthenozoospermia group (41.24% ± 9.71%) was significantly lower than that in control group (56.68% ± 11.13%). MMP was negatively correlated with BMI (r = −0.25, P < 0.01), but positively correlated with total sperm motility (r = 0.63, P < 0.01), motility of progressive sperm (r = 0.64, P < 0.01), and normal sperm morphology rate (r = 0.37, P < 0.01). In addition, MMP showed no significant correlations with age, volume of semen, sperm concentration, sperm count, and other indexes. Conclusions: Sperm MMP is an important index in the evaluation of sperm function, and detection of MMP may provide references for the diagnosis and treatment of male infertility. Key words: Body Mass Index; Conventional Semen Analyses; JC-1; Mitochondrial Membrane Potential

Micrometastasis Detection using Special Stains in Nodal Tissues of Oral Squamous Cell Carcinoma - A Histochemical Study.

Micrometastasis to the lymph node is an important prognostic factor in survival and recurrence and even in making critical decisions regarding postoperative radiation treatment. Methods like Immunohistochemistry (IHC), serial sectioning and Polymerase Chain Reaction (PCR) are very sensitive; however, they are expensive and time consuming. Hence, there is a need for the detection of micrometastasis by method which is easily feasible and inexpensive. This study was undertaken to identify if special stains (Modified Papanicolaou stain and Ayoub Shklar stain) are valuable in detecting micrometastasis which are routinely missed in Haematoxylin and Eosin stain and also to compare their efficacy over Haematoxylin and Eosin stain in detection of metastatic tumour cells in non-metastatic lymph nodes. This study constituted a total of 300 histopathologically proven non metastatic lymph node sections. Other than Haematoxylin and Eosin stain, Modified Papanicalaou and Ayoub Shklar stain were used for identification of micrometastatic deposit. Papanicolaou stain was the only stain useful in detecting micrometastasis which accounted to 7% of non metastatic lymph node sections used in our study. Papanicolaou stain has definitely proved valuable in detecting micrometastasis over routine Haematoxylin and Eosin and Ayoub Shklar stain with an added advantage of being economical, easily available and technique insensitive.

A Novel Ovine Primary Alveolar Type II Cell Culture Approach

The in vivo ovine model provides a clinically relevant platform to study cardiopulmonary development; however, a developmental ovine primary ATII cell culture model is lacking. The objective of this study to develop and optimize submerged and air-liquid interface ovine ATII cell culture methodologies for the purposes of dissecting mechanisms at the cellular level to elucidate responses observed in vivo. Lung tissue was harvested from mechanically ventilated preterm (128-132 d gest.), juvenile (4-6 months), adult (2-4 yrs) sheep and then immediately processed or frozen until processing for 7 day cell culture methods: submerged (7 d) or submerged (3 d) followed by air-liquid interface cell culture (4 d). Cells viability (>95%) was determined by Trypan blue exclusion; isolated ATII cell purity (>90%) was determined by modified Papanicolaou stain, ATII cells were identified by the presence of lamellar bodies, and ATII yield was normalized per gram tissue (preterm: 2.6 x 106; juvenile: 1.9 x 106; adult: 0.70 x 106). Presence, abundance, and mRNA expression of surfactant protein B (SP-B) was assessed by immunocytochemistry, Western Blot, and quantitative PCR, respectively, on the day of isolation, at midpoint and at the end of the study period. In cells derived from all age-groups from either fresh or frozen tissue, all biomarkers of SP-B were significantly greater in air-liquid interface as compared to submerged culture conditions at all time points. These data demonstrate an optimized cell culture methodology using fresh or frozen tissue to support study of ovine primary ATII cell function and responses to further understanding of mechanisms that contribute to in vivo function of the lung. DOD/ONR;N000141210597;N000141210810.

Cytomorphometric Analysis of Oral Buccal Mucosal Smears in Tobacco and Arecanut Chewers Who Abused With and Without Betel Leaf

ABSTRACT. Background: Tobacco in any form (smoking or chewing), arecanut chewing, and alcohol are considered to be the major extrinsic etiological factors for potentially malignant disorders of the oral cavity and for squamous cell carcinoma, the most common oral malignancy in India. An increase in nuclear diameter (ND) and nucleus-cell ratio (NCR) with a reduction in cell diameter (CD) are early cytological indicators of dysplastic change. The authors sought to identify cytomorphometric changes in ND, CD, and NCR of oral buccal cells in tobacco and arecanut chewers who chewed with or without betel leaf. Methods: Participants represented 3 groups. Group I consisted of 30 individuals who chewed tobacco and arecanut with betel leaf (BQT chewers). Group II consisted of 30 individuals who chewed tobacco and arecanut without betel leaf (Gutka chewers). Group III comprised 30 apparently healthy nonabusers. Cytological smears were prepared and stained with modified-Papanicolaou stain. Results: Comparisons between Groups I and II and Groups II and III showed that ND was increased, with P values of .054 and .008, respectively, whereas a comparison of Groups I and III showed no statistical significance. Comparisons between Groups I and II and Groups II and III showed that CD was statistically reduced, with P values of .037 and <.000, respectively, whereas comparison of Groups I and III showed no statistical significance. Comparisons between Groups I and II and groups II and III showed that NCR was statistically increased, with P values of <.000, whereas a comparison of Groups I and III showed no statistical significance. Conclusions: CD, ND, and NCR showed statistically significant changes in Group II in comparison with Group I, which could indicate larger and earlier risk of carcinoma for Gutka chewers than in BQT chewers.

Analytical Validation of the ForeCYTE Breast Health Test

Introduction: The ForeCYTE Breast Health Test provides a 10-year and lifetime breast cancer risk stratification using family history of breast and ovarian cancer, personal reproductive history, and cytological evaluation of exfoliated cells present in nipple aspirate fluid (NAF) in the risk stratification algorithm developed by TyrerCuzick. Materials and Methods: Specimens of NAF are collected on membrane filters with one of two FDA-cleared biopsy instruments, the Halo or the Mammary Aspiration Specimen Cytology Test (MASCT) System devices, are fixed upon collection by spraying the membrane filters with a 0.1 mL mist of Saccomanno's fixative. The specimen filters are processed in situ for NAF protein using a colloidal gold assay with a limit of detection of 0.1 ng of protein (corresponding to 1 pL of NAF) and a linear range to 1,000 ng. The filters are then stained with a modified Papanicolaou stain and morphological criteria applied to establish a diagnosis. Results: Spiked MCF-7 breast cancer cells demonstrate quantitative recovery (94.7%) in the range of 1-10 cancer cells/membrane filter with a sensitivity for identifying a cellular specimen of 100% at 4-100 cells/membrane and 75% at 2 cells/membrane and 50% at 1 cell /membrane. If hyperplasia is noted and morphology is inadequate

Cystic fibrosis transmembrane conductance regulator is correlated closely with sperm progressive motility and normal morphology in healthy and fertile men with normal sperm parameters

Cystic fibrosis transmembrane conductance regulator (CFTR) has been demonstrated to be expressed in mature spermatozoa and correlated with sperm quality. Sperm CFTR expression in fertile men is higher than that in infertile men suffering from teratospermia, asthenoteratospermia, asthenospermia and oligospermia, but it is unknown whether CFTR is correlated with sperm parameters when sperm parameters are normal. In this study, 282 healthy and fertile men with normal semen parameters were classified into three age groups, group (I): age group of 20-29 years (98 cases, 27.1 ± 6.2), group (II): age group of 30-39 years (142 cases, 33.7 ± 2.6) and group (III): age group of more than or equal to 40 years (42 cases, 44.1 ± 4.6). Sperm concentration, total count and progressive motility were analysed by computer-assisted sperm analysis. Sperm morphology was analysed by modified Papanicolaou staining. Sperm CFTR expression was conducted by indirect immunofluorescence staining. There was a significant positive correlation (P < 0.001) between CFTR expression and sperm progressive motility (r = 0.221) and normal morphology (r = 0.202), but there were no correlations between sperm CFTR expression and semen volume, sperm concentration, sperm total count as well as male age (P > 0.05). Our findings show that CFTR expression is associated with sperm progressive motility and normal morphology in healthy and fertile men with normal sperm parameters, but not associated with the number of spermatozoa and male age.

GW24-e1279 Sperm DNA Fragmentation increased in Males with Metabolic Syndrome

ObjectivesTo analyse the sperm DNA fragmentation and sperm morphology in males with metabolic syndrome and healthy men. Methods28 males with metabolic syndrome (mean age 33.64 ± 7.42 years), according to...

Comparing modified papanicolaou stain with ayoub-shklar and haematoxylin-eosin stain for demonstration of keratin in paraffin embedded tissue sections

Aim:The aim of the present study was to stain the known keratin containing tissues by Haematoxylin and eosin stain (H-E), ayoub-shklar (A/S) and modified Papanicolaou (PAP) stain and to compare the efficacy of modified PAP staining procedure with that of A/S stain and H-E staining technique, so as to device a staining procedure which is easy and effective for keratin.Materials and Methods:A Total Number of 60 paraffin embedded tissue sections of known keratin containing tissues including normal keratinized oral mucosa (NKOM), Keratinized Odontogenic Keratocyst (OKC), Verrucous Carcinoma (VC) and Well differentiated Squamous Cell Carcinoma (WDSCC) were taken and 3 sections of 4 microns thickness of each block were cut and stained with above mentioned three stains.Results:Surface keratin was stained distinctly and uniformly in all the three staining techniques in NKOM, OKC, and VC and WDSCC. But results were statistically significant in WDSCC when Amount of keratin pearls and Pattern of staining were compared in all 3 staining procedures and p value was p=0.000 and p=0.001 respectively.Conclusion:Based on the above findings we conclude that the efficacy of modified PAP is comparable with that of H-E stain and A-S stain for surface keratin and thus be used effectively to stain Surface keratin. Whereas to know the exact pattern of cytokeratin expression in SCC a more sensitive tool like immunohistochemical method can be applied.

Efficacy of Modified Papanicolaou stain for demonstration of keratin in paraffin embedded tissue sections

Aims and Objectives The aim of the present study was to stain the known keratin containing paraffin embedded tissues by Haematoxylin and Eosin stain H-E and Modified Papanicolaou PAP stain and to compare the efficacy of modified PAP staining procedure with that of H-E staining technique so as to device a staining procedure which is easy and effective for keratin. Materials and methods A Total Number of 40 paraffin embedded tissue sections of known keratin containing tissues including Normal Keratinized Oral Mucosa NKOM Keratinized Odontogenic Keratocyst OKC and Verrucous Carcinoma VC were taken and 2 sections of 4micron thickness of each block were cut and stained with above mentioned two stains. Results were analyzed using Chi square test where P value le0.05 was considered to be statistically significant. Results Surface keratin was stained distinctly and uniformly in both staining techniques when analysed for Type of surface keratin and Pattern of surface keratin.Conclusion Based on the above findings we conclude that the efficacy of modified PAP is comparable with that of H-E stain for Surface keratin and thus can be used effectively to stain Surface keratin in paraffin embedded tissue sections.nbsp

Comparison of the quality of smears in transbronchial fine‐needle aspirates using two staining methods for rapid on‐site evaluation

Transbronchial needle aspiration (TBNA) via flexible bronchoscopy is a well-established sampling modality for lung masses. The procedure is useful in the diagnosis of neoplastic and non-neoplastic lesions as well as for staging of bronchogenic carcinoma. Rapid on-site evaluation (ROSE) adds value as it has the advantage of triaging material during the procedure so avoiding a battery of investigations. Frequently used rapid stains are the modified Wright-Giemsa water-based stain (WG-ROSE) and the alcohol-based modified Papanicolaou stain (Pap-ROSE). Final review of laboratory-based Giemsa and Pap stains supplemented by ancillary investigations is essential for quality assurance. To investigate whether and how ROSE influenced the quantity and quality of the material submitted to the laboratory we randomized 126 patients to WG-ROSE, requiring only one pathologist on-site, or combined WG- and Pap-ROSE, requiring an additional person on-site to assist with staining. In those patients with positive TBNA we graded the laboratory-based slides of the first pass containing diagnostic material into insufficient, suspicious, adequate and excellent. The first diagnostic pass was found after 3.06 ± 1.94 (SD) passes and 3.13 ± 2.16 passes with WG-ROSE and combined ROSE (P = 0.87), respectively. Following WG-ROSE and combined ROSE 69% and 71.1% (P = 0.509) of slides were diagnostic (adequate or excellent) on laboratory-based Giemsa stains, and 93.3% and 100% (P = 0.134) were scored adequate or excellent on laboratory-based Pap stains. We concluded that the less costly and labour intensive WG-ROSE procedure is adequate for TBNA. This has cost implications especially in resource poor settings.

Modified Papanicolaou staining protocol with minimum alcohol use: a cost‐cutting measure for resource‐limited settings

To devise a simple, cost-effective protocol for Papanicolaou (Pap) staining of cervicovaginal smears. Five hundred coded paired cervical smears were collected from women as part of routine cervical cancer screening. One set of smears was stained by conventional Pap staining protocol (CP) and the other by a modified protocol (MP) in which alcohol was replaced by 1% acetic acid in all the steps except during fixation and prior to mounting; in addition, one alcohol-based counterstain, OG, was omitted. The smears were examined blindly by the pathologists and then decoded. Each pair of smears was compared and the two protocols were analysed for staining quality and diagnoses by McNemar and chi-square tests. The staining quality in the MP was satisfactory. The nuclear and cytoplasmic features were comparable to the CP. Cytoplasmic transparency was maintained in the MP and the differential staining of blue/green and pink was acceptable to the pathologists and technicians. The diagnoses agreed in all cases and there was no compromise in interpreting the smears. With MP it took only 3-4 minutes to stain a batch of 50 slides. in contrast to the 20 minutes taken by CP. The MP used almost one-seventh of the amount of alcohol compared with CP, which translated into a significant cost reduction per smear. The improvised Pap staining protocol with minimum alcohol use is a simple, cost-effective and technician-friendly procedure that can be easily adopted in high-volume, resource-limited laboratories for mass cervical cancer screening.

O166. Lab-on-a-chip sensor for analysis of cellular biomarkers in oral exfoliative cytology: A new diagnostic tool for early detection of oral cancer

Introduction: Analysis of cytological samples from potentially malignant oral lesions (PMOLs) can be of diagnostic and possibly prognostic value to the clinician. Objective: To compare the performance, on cytological samples procured from subjects with known SCCA or PMOLs, of quantitative cytology (Perceptronix, Canada) versus the Oral CDX Brush Test (CDX, USA) against matched histopathology (‘‘gold standard”). Methods: Subjects with known SCCA or PMOLs (categorized into low or high risk of clinical suspicion) underwent an oral brush biopsy, whereupon a cytological sample was plated and fixed. Instead of discarding the brushes, the tips with residual cells were retained and fixed. Unless a known SCCA, a scalpel biopsy was immediately performed at the brush site and tissue sent for histopathology. Glass slides were sent to CDX for modified Papanicolaou staining and automated cytological analysis, and reported by a cytopathologist as normal (CDX negative) or abnormal (CDX positive or atypical). Leftover brushes were sent to Perceptronix, cytospun, underwent Feulgen-Thionin staining, and DNA ploidy analysis using the ClearCyte automated image cytometry system. Galleries containing nuclei with non-diploid DNA content were reviewed by a cytopathologist, and reported as normal or abnormal. Results: A total of 86 cytological samples were of sufficient yield to undergo both tests. Of these samples, 57 (66%) had a diagnosis of dysplasia or SCCA. Compared with the ‘‘gold standard”, the sensitivity/specificity/PPV/NPV of quantitative cytology versus the Brush Test in detecting dysplasia (any grade) or SCCA were 61/100/100/ 57% versus 77/83/90/65%, respectively. Overall performance increased on high risk suspicion PMOLs and when detecting higher grade dysplasias or SCCA. Quantitative cytology had additional value in resolving CDX atypical results. Conclusion: Each test offers an advantage and disadvantage over the other. Longitudinal studies are needed to assess the utility of such tests in low risk PMOLs or low grade dysplasias. Funded by NYUCD Dean’s Student Research Award & NIDCR/NIH Grant # U54-DE14257.

Imprint cytology of vacuum‐assisted breast biopsy specimens: a rapid diagnostic tool in non‐palpable solid lesions

Imprint cytology provides a rapid preliminary diagnosis shortly after the completion of breast biopsy. This study aims to assess the validity of imprint cytology for the pre-operative diagnosis of non-palpable mammographic solid lesions excised by vacuum-assisted breast biopsy (VABB). Seventy-two women with non-palpable Breast Imaging Reporting and Data System 3 and 4 mammographic solid lesions without microcalcifications underwent VABB on the stereotactic Fischer's table with 11-G Mammotome vacuum probes. Imprint samples were examined (Diff-Quick stain, modified Papanicolaou stain and May-Grünwald-Giemsa). The cores were dipped into a CytoRich Red Collection fluid for a few seconds in order to obtain samples with the use of the specimen wash. After the completion of cytological procedures, the core was prepared for routine pathological study. The pathologist was blind to the preliminary cytological results. The cytological and pathological diagnoses were comparatively evaluated. The sensitivity of the cytological imprints for cancer was 90%. The specificity of the method for cancer diagnosis was 100%. Two precursor lesions were present in the material: one case of atypical ductal hyperplasia, which was successfully detected, and one case of lobular neoplasia, which escaped detection. The cytological imprints were inadequate in four out of 72 cases (5.6%), but none of them were included within the malignant subgroup. Imprint cytology seems to be an important adjunctive tool in the management of patients with non-palpable mammographic solid lesions. Its very satisfactory sensitivity and optimal specificity could establish its use in general clinical practice.

Annexin A2 Interactions with Rab14 in Alveolar Type II Cells

Annexin A2, a calcium-dependent phospholipid-binding protein, is abundantly expressed in alveolar type II cells where it plays a role in lung surfactant secretion. Nevertheless, little is known about the details of its cellular pathways. To identify annexin A2-regulated or associated proteins, we silenced endogenous annexin A2 expression in rat alveolar type II cells by RNA interference and assessed the change of the cellular transcriptome by DNA microarray analysis. The loss of annexin A2 resulted in the change of 61 genes. Thirteen of the selected genes (11 down-regulated and 2 up-regulated genes) were validated by real time quantitative PCR. When the loss of rat annexin A2 was rescued by overexpressing EGFP-tagged human annexin A2, six of seven selected targets returned to their normal expression level, indicating that these genes are indeed annexin A2-associated targets. One of the targets, Rab14, co-immunoprecipitated with annexin A2. Rab14 also co-localized in part with annexin A2 and lamellar bodies in alveolar type II cells. The silencing of Rab14 resulted in a decrease in surfactant secretion, suggesting that Rab14 may play a role in surfactant secretion.

Imprint cytology on microcalcifications excised by vacuum-assisted breast biopsy: a rapid preliminary diagnosis.

BackgroundTo evaluate imprint cytology in the context of specimens with microcalcifications derived from Vacuum-Assisted Breast Biopsy (VABB).Patients and methodsA total of 93 women with microcalcifications BI-RADS 3 and 4 underwent VABB and imprint samples were examined. VABB was performed on Fischer's table using 11-gauge Mammotome vacuum probes. A mammogram of the cores after the procedure confirmed the excision of microcalcifications. For the application of imprint cytology, the cores with microcalcifications confirmed by mammogram were gently rolled against glass microscope slides and thus imprint smears were made. For rapid preliminary diagnosis Diff-Quick stain, modified Papanicolaou stain and May Grunwald Giemsa were used. Afterwards, the core was dipped into a CytoRich Red Collection fluid for a few seconds in order to obtain samples with the use of the specimen wash. After the completion of cytological procedures, the core was prepared for routine histological study. The pathologist was blind to the preliminary cytological results. The cytological and pathological diagnoses were comparatively evaluated.ResultsAccording to the pathological examination, 73 lesions were benign, 15 lesions were carcinomas (12 ductal carcinomas in situ, 3 invasive ductal carcinomas), and 5 lesions were precursor: 3 cases of atypical ductal hyperplasia (ADH) and 2 cases of lobular neoplasia (LN). The observed sensitivity and specificity of the cytological imprints for cancer were 100% (one-sided, 97.5% CI: 78.2%–100%). Only one case of ADH could be detected by imprint cytology. Neither of the two LN cases was detected by the imprints. The imprints were uninformative in 11 out of 93 cases (11.8%). There was no uninformative case among women with malignancy.ConclusionImprint cytology provides a rapid, accurate preliminary diagnosis in a few minutes. This method might contribute to the diagnosis of early breast cancer and possibly attenuates patients' anxiety.