Inter-observer reproducibility of endometrial cytology by the Osaki Study Group method: utilising the Becton Dickinson SurePath™ liquid-based cytology.

OBJECTIVE: The purpose of the present study was to evaluate the reproducibility of the cytological diagnosis of endometrial lesions by the Osaki Study Group (OSG) method of new cytological diagnostic criteria using BD SurePath™ (SP)-liquid-based cytology (LBC). METHODS: This cytological classification using the OSG method consists of six categories: (i) normal endometrium (NE), (ii) endometrial glandular and stromal breakdown (EGBD), (iii) atypical endometrial cells, cannot exclude atypical endometrial hyperplasia or more (ATEC-A), (iv) adenocarcinoma including atypical endometrial hyperplasia or malignant tumour (Malignancy), (v) endometrial hyperplasia without atypia (EH) and (vi) atypical endometrial cells of undetermined significance (ATEC-US). For this study, a total 244 endometrial samplings were classified by two academic cytopathologists as follows: 147 NE cases , 36 EGBD cases , 47 Malignant cases, eight ATEC-A cases, two EH cases and four ATEC-US cases. To confirm the reproducibility of the diagnosis and to study the inter- and intra-observer agreement further, a second review round followed at 3-month intervals, which included three additional cytopathologists. RESULTS: The inter-observer agreement of NE classes improved progressively from 'good to fair' to 'excellent', with values increasing from 0.70 to 0.81. Both EGBD and Malignancy classes improved progressively from 'good to fair' to 'excellent', with values increasing from 0.62-0.63 to 0.84-0.95, respectively. The overall intra-observer agreement between the first and the second rounds was 'good to fair' to 'excellent', with values changing from 0.79 to 0.85. All kappa improvements were significant (P < 0.0001). CONCLUSION: In this study, it seemed that the use of the OSG method as the new diagnostic criteria for SP-LBC preparation, may be a valid method to improve the precision (reproducibility) of endometrial cytology.

Body cavity fluid can induce epithelial and mesothelial differentiation from CD34 positive peripheral blood stem cells in vitro.

OBJECTIVE: Primary culture of CD34 positive stem cells collected from human peripheral blood was performed with and without supplementation with concentrated ascitic fluid; morphological and immunocytochemical pictures of cultured cells were taken chronologically and compared. METHODS: CD34-positive stem cells collected from peripheral blood were cultured for 1, 24 and 48 hours. Concentrated ascitic fluid was added to the plates for the 24-and 48-hour cultures. For immunocytochemical studies, CD34, AE1/AE3, Ber-Ep4 (EA), EMA, EGFR, CD31, CA125 and D2-40 monoclonal antibodies were used. RESULTS: After culture, small round cells with naked nuclei began to enlarge and to exhibit various changes in the cytoplasm and nucleus. Supplementation with concentrated body cavity fluid enhanced these changes. CD34-positive cells with small round cell features were detected 1 hour after culture and these had no epithelial or mesothelial markers. After 24 hours, CD34-positive cells had disappeared and cells weakly positive for EGFR, EMA, CA125 and D2-40 were detected. Cells with strong and moderate positive reactions for EGFR, AE1/AE3, EA, EMA, D2-40 and CA125 were detected after 48 hours. Supplementation with concentrated body cavity fluid increased the intensity and number of positive cells for these markers compared with the control group. The positive reaction, not only for the epithelial markers such as EGFR and AE1/AE3, but also for mesothelial markers such as CA125 and D2-40, was found to be increased in small numbers of cells in direct proportion to the duration of the primary culture of the peripheral blood cells. CD31, characteristically expressed in endothelial cells, was negative in the cultured cells. CONCLUSION: Supplementation of peripheral blood CD34-positive stem cells with body cavity fluid in vitro enhanced their differentiation toward cells of an epithelial or mesothelial phenotype, concomitant with loss of immunoreactivity for CD34. It is assumed that the routine cytological observation of cells obtained from body cavity fluid might cause possible cytomorphological and immunophenotypical changes due to the action of the growth factors contained in the body cavity fluid.

Conceptual design of heavy ion beam probes on the PLATO tokamak.

Heavy ion beam probe (HIBP) systems have been designed for the new tokamak, PLATO [A. Fujisawa, AIP Conf. Proc. 1993, 020011 (2018)]. The designs have been completed, and the installations are in progress. Two HIBPs are being installed in toroidal sections 180° apart to investigate long-range correlations in the toroidal direction. Each HIBP consists of an injection beamline and a detection beamline as usual. Yet, one of the HIBPs is equipped with an additional detection beamline; the measurement positions of its two detection beamlines can be placed on almost the same magnetic surface yet at poloidal angles that differ by ∼180°. The use of three detection beamlines allows us to investigate spatial asymmetry and long-range correlations in both the toroidal and poloidal directions, simultaneously. The detected beam intensity is expected to be enough for turbulence measurements in almost the entire plasma region when the electron density is up to 1 × 1019 m-3 by selecting appropriate ion species for the probe beam. Each detector has three channels 10 mm apart, allowing measurement of local structures of micro-scale turbulence. Therefore, using the HIBPs on the PLATO tokamak will enable both local and global properties of plasma turbulence to be investigated, simultaneously.

EMMPRIN (CD147) expression and differentiation of papillary thyroid carcinoma: implications for immunocytochemistry in FNA cytology.

OBJECTIVE: Extracellular matrix metalloproteinase inducer (EMMPRIN) and its induced matrix metalloproteinases (MMPs) play a crucial role in tumour progression, invasion and metastasis. EMMPRIN expression has been demonstrated in several tumours, but its expression profile in thyroid cancer remains unclear. METHODS: We evaluated the expression profile of EMMPRIN at various stages of differentiation of thyroid carcinoma, including 20 cases of well-differentiated papillary carcinoma (WDPC), 15 cases of papillary carcinoma with a poorly differentiated carcinoma component (PC/PDC) and four cases with an undifferentiated carcinoma (UDC) component, using paraffin-embedded sections for immunohistochemical stains. Also, we used 32 fine needle aspiration cytology and imprint smears from the same cases for immunocytochemical stains. The staining results were evaluated with a scoring system. RESULTS: Immunohistochemical staining showed that EMMPRIN expression was absent or weak in almost all WDPC specimens, whereas it was moderate or strong in PDC and UDC components. In tumours that showed a gradual morphological transformation from WDPC to PDC components, the expression of EMMPRIN was progressively stronger from the areas of WDPC to those of PDC. WDPC, PC/PDC and UDC had expression scores of 4.9, 45.0 and 245.7, respectively. Results of immunocytochemical staining showed almost the same staining profile as those of immunohistochemical staining. The cytological atypia of EMMPRIN-positive cells was greater than that of negative cells. CONCLUSION: These results indicated that EMMPRIN expression correlates significantly with the degree of dedifferentiation of thyroid carcinoma. This study demonstrates the feasibility of expression of EMMPRIN using fine needle aspiration samples. Therefore, immunocytochemical analysis of EMMPRIN may be a novel aid to evaluate the differentiation of thyroid carcinoma.

Survey of medical training in cytopathology carried out by the journal Cytopathology.

This report of the Editorial Advisory Board of Cytopathology gives the results of a survey of medical practitioners in cytopathology, which aimed to find out their views on the current situation in undergraduate and postgraduate training in their institutions and countries. The results show that training in cytopathology and histopathology are largely carried out at postgraduate level and tend to be organized nationally rather than locally. Histopathology was regarded as essential for training in cytopathology by 89.5% of respondents and was mandatory according to 83.1%. Mandatory cytopathology sections of histopathology were reported by 67.3% and specific examinations in cytopathology by 55.4%. The main deficiencies in training were due to its variability; there were insufficient numbers of pathologists interested in cytology and a consequent lack of training to a high level of competence. Pathologists without specific training in cytopathology signed out cytology reports according to 54.7% of responses, more often in centres where training was 3-6 months or less duration. Although 92.2% of respondents thought that specialist cytology should not be reported by pathologists without experience in general cytopathology, that practice was reported by 30.9%, more often in centres with small workloads. The survey report recommends that 6-12 months should be dedicated to cytopathology during histopathology training, with optional additional training for those wanting to carry out independent practice in cytopathology. Formal accreditation should be mandatory for independent practice in cytopathology. When necessary, temporary placements to centres of good practice should be available for trainees intending to practise independently in cytopathology. There should be adequate numbers of pathologists trained in cytopathology to a high level of competence; some of their time could be released by training cytotechnologists and trainee pathologists to prescreen cytology slides and assess adequacy of fine-needle aspiration samples when immediate diagnosis was not required. The survey demonstrated a clear need for European and international guidelines for training in cytopathology.

Utility of liquid-based cytology in endometrial pathology: diagnosis of endometrial carcinoma.

OBJECTIVE: The purpose of this study was to examine the utility of SurePath-liquid-based cytology (LBC) compared to conventional cytological preparations (CCP) in the identification of endometrial carcinoma. METHODS: During a 13-month period, direct endometrial samples were collected from 120 patients using the Uterobrush. The material comprised 30 cases each of endometrial carcinoma, proliferative endometrium, secretory endometrium and atrophic endometrium. The following points were investigated:(i) the frequency of cell clumps in endometrial carcinoma; (ii) the area of cell nuclei; (iii) overlapping nuclei. RESULTS: (i) Comparison of the frequency of cell clumps with irregular protrusion pattern and papillo-tubular pattern showed no statistically significant difference in either type of cell clump between CCP and LBC. (ii) Comparison of the nuclear area of cells showed a sequential decrease from endometrial carcinoma to secretory endometrium, to proliferative endometrium and to atrophic endometrium, which was significant in CCP and LBC. (iii) Nuclear area was significantly lower with LBC compared with CCP in endometrial carcinoma, secretory endometrium and proliferative endometrium but not atrophic endometrium. (iv) Comparison of the degree of overlapping nuclei showed a sequential decrease from endometrial carcinoma to proliferative endometrium, to secretory endometrium and to atrophic endometrium, which was significant in both CCP and LBC. (v) Comparison of the degree of overlapping nuclei between CCP and LBC showed no significant difference for normal types of endometrium, but LBC had significantly higher values (P < 0.0001) in endometrial carcinoma than in CCP. CONCLUSIONS: The results of this study revealed that applying diagnostic criteria used in CCP to LBC was easy to achieve, because LBC had excellent cytoarchitectural preservation and cells were well presented. Although we have not examined all cytological features of malignancy and have not considered atypical hyperplasia, we believe that this method may be a useful tool in the diagnosis of endometrial cytology.

New diagnostic reporting format for endometrial cytology based on cytoarchitectural criteria.

OBJECTIVE: The aim of this study was to develop a new reporting format for endometrial cytology that would standardize the diagnostic criteria and the terminology used for reporting. METHODS: In previous studies, cytoarchitectural criteria were found to be useful for the cytological assessment of endometrial lesions. To apply these criteria, an appropriate cytological specimen is imperative. In this article, the requirements of an adequate endometrial cytological specimen for the new diagnostic criteria are first discussed. Then, the diagnostic criteria, standardized on a combination of conventional and cytoarchitectural criteria, are presented. Third, terminology that could be used, not only for reporting the histopathological diagnosis, but also for providing better guidance for the gynaecologist to determine further clinical action, is introduced. The proposed reporting format was investigated using endometrial cytology of 58 cases that were cytologically underestimated or overestimated compared to the histopathological diagnosis made on the subsequent endometrial biopsy or surgical specimens. RESULTS: Of the 58 cases, 12 were reassessed as being unsatisfactory for evaluation. Among the remaining 46 cases, 25 of the 27 cases, which had been underestimated and subsequently diagnosed as having endometrial carcinoma or a precursor stage on histopathological examination,were reassessed as recommended for endometrial biopsy. On the other hand, 19 cases overestimated by cytology were all reassessed as not requiring biopsy. CONCLUSIONS: The reporting format for endometrial cytology proposed in this article may improve diagnostic accuracy and reduce the number of patients managed inappropriately.

Cytopathology of pregnancy-induced cell patterns in cervicovaginal smears.

Considerable interest has been devoted to cytology in pregnancy, especially the morphologic changes that may cause problems in differential diagnosis. It is surprising that the published discussion of the cytologic appearance of smears from pregnant women has been so limited. This review emphasizes that retained trophoblastic tissue may be a source of highly atypical appearing cells in the cervicovaginal cytology obtained under various clinical conditions. Distinguishing between Arias-Stella cells and cells of glandular abnormalities can be problematic, since the morphologic characterization of the former is poor. This review also emphasizes that a full awareness of the morphology of pregnancy as well as of the patient's clinical history are needed for greater precision in diagnosing cell patterns as pregnancy-related and not malignant.

Cytologic evaluation of conjunctival epithelium after cataract surgery.

Although numerous studies have examined corneal epithelial wound healing, few reports describe conjunctival wound healing. Thus, we evaluated the conjunctival epithelium by brush cytology, in which a miniature brush was used to collect many cells relatively noninvasively. The temporal and superior conjunctivae were examined before and after cataract extraction on the first, third, seventh, and fourteenth days and then the first, second, and third months postoperatively, in a group of 20 patients (9 men and 11 women with a mean age of 69 years 3 months +/- 10 years). The cells collected were stained by the Papanicolaou method. Each postoperative specimen contained keratinized, polygonal, round, columnar, elongated, and inflammatory cells, as well as cells with nuclear alterations. No keratinized, elongated, or inflammatory cells were observed preoperatively. In contrast, columnar cells, which were present preoperatively, decreased during the wound healing process. Such nuclear changes as chromatin formation or the appearance of snake-like figures were observed in several patients postoperatively. These cellular changes may represent altered differentiation during the healing process. However, the conjunctival epithelium returned to normal within 2 months postoperatively in all patients.

The cellular composition of uterine fluid in IUD users - a quantitative study.

The cellular composition of uterine fluid was studied quantitatively in 75 IUD users and 48 non-users. In the IUD group the number of granulocytes was increased about 300 times above that of the non-IUD group, whereas the numbers of mononuclear phagocytic cells and lymphocytes were increased by a factor of 30. Endometrial glandular cells were not quantitatively affected by the presence of an IUD. Granulocytes constituted nearly two-thirds and mononuclear phagocytic cells nearly one-third of the total cell count in the IUD group. Mast cells wee found more frequently and eosinophilia occurred more often in the uterine fluid of IUD users.

Comparison of immunocytochemistry and in situ hybridization in the cytodiagnosis of genital herpetic infection.

Over a 62-month period, 53 patients were found to have cervicovaginal smears that contained cells consistent with, or equivocal cells for, a herpes simplex virus (HSV) infection. The Papanicolaou-destained smears from these cases were restrained in situ hybridization (ISH) with a biotinylated cloned DNA probe and immunocytochemistry (ICC) assay and were compared for the detection of HSV in cervicovaginal smears by two methods. Cytological findings classified the 53 slides into two groups, i.e., cytologically herpes positive (33 patients) and equivocal cases (20 patients). Each group was subdivided into two groups: group A was confirmed by ICC, and group B was confirmed by ISH technique. Of the 33 cellular samples containing cells considered to be consistent with a herpes infection, 15 (88%) of 17 were positive by means of ICC technique (group A), 6 (43%) of 14 were positive by ISH technique (group B). Of the 20 smears showing equivocal cell changes thought unlikely to be caused by an HSV infection, 6 (60%) of 10 were positive by ICC (group A), 2 (29%) of 7 were positive by ISH (group B). With the ISH technique, five smears showed dislodged cells from glass slides due to enzyme treatment and denaturation. The results revealed that the ICC technique is a rapid and reliable procedure and thus recommended for routine diagnosis of HSV infection. Moreover, ICC is easier to perform and interpret and is less expensive than ISH. Therefore, the ICC may be preferable to ISH for detecting HSV in routine Papanicolaou diagnostic work.

Gastric carcinoma metastatic to the bone marrow: immunoperoxidase identification of KMO-1 antigen in MGG-destained aspirate.

A case is presented that illustrates the application of the immunoperoxidase technique to the May-Grünwald-Giemsa (MGG)-destained bone marrow aspirate. The cytologic findings in a MGG-stained smear of the bone marrow suggested a metastatic epithelial tumor. Subsequently, a positive reaction to KMO-1, a monoclonal antibody raised against a colon carcinoma cell line, was demonstrated in tumor cells in the MGG-destained smear sample as well as in the paraffin-embedded section of the primary gastric cancer. The demonstration of the cancer-associated antigen in the MGG-destained material may be useful in establishing the diagnosis of metastatic tumor in the bone marrow.

Immunocytochemical localization of carcinoembryonic antigen in cerebrospinal fluid with metastatic carcinoma of the stomach: report of four cases.

A study was undertaken to evaluate the cellular findings of gastric carcinoma in cerebrospinal fluid (CSF). Immunocytochemical localization of carcinoembryonic antigen (CEA) was performed on four cases of metastatic gastric carcinoma cells in CSF samples. A positive peroxidase-antiperoxidase (PAP) reaction was obtained in all cases, with intense staining for CEA in the CSF samples as well as in the paraffin-embedded tissue sections from the primary gastric tumors. Cellular morphology and the results of immunoperoxidase staining can be studied simultaneously. We believe that the PAP method for CEA increases diagnostic accuracy of cytology in the CSF.

Cytologic detection of herpes simplex virus DNA in nipple discharge by in situ hybridization: report of two cases.

Cytologic changes in the smears from nipple discharge of two cases with herpes simplex virus (HSV) infections are described. The cytology revealed the ground-glass appearance of the nuclei with multinucleated syncytial cells. Subsequently, in situ hybridization (ISH) using a biotinylated DNA probe was applied to identify the HSV DNA in the Papanicolaou-destained specimens. Positive hybridization was found with intense staining for the HSV DNA in the nuclei of cells having a ground-glass appearance. Cytologic observation together with ISH procedure may be rapid and valuable tool for the detection and final demonstration of HSV infections. Similar investigations may be carried out in cellular samples from other body sites.

Immunocytochemical presentation of alpha-fetoprotein-producing gastric cancer in ascitic fluid: a case study.

A case of primary gastric cancer without hepatic metastasis showing extremely high alpha-fetoprotein (AFP) levels is reported. This case illustrates the application of the immuno-peroxidase technique to ascitic fluid cytology. Papanicolaou-stained smears of the ascites permitted the diagnosis of a metastatic carcinoma. A positive reaction to AFP was demonstrated in the tumor cells in the ascitic fluid cellular samples as well as in the paraffin-embedded tissue section of the primary gastric carcinoma. Rising AFP levels were also detected in ascitic fluid. AFP fractionation using lectin-affinity-crossed-line immunoelectrophoresis showed the hepatic rather than yolk sac type. Reports of such occurrences are few; no study, to the best of our knowledge, has previously documented cytological and immunocytochemical diagnosis in ascitic fluid. AFP-producing gastric cancer should be considered in the differential diagnosis.

Ciliated ovarian adenocarcinoma cells in ascitic fluid cytology: report of a case with immunocytochemical features.

A case of ovarian serous adenocarcinoma presenting with ascitic fluid is reported. Cytology of the ascitic fluid showed the presence of cilia on most of malignant tumor cells. These tumor cells occurred singly; cilia usually appeared in unipolar locations. Immunocytochemical studies were performed on the ascitic fluid samples as well as tissue sections. CA19-9, CA125, CA50, and epithelial antigens were positive, while carcinoembryonic antigen was negative. The positive reactions were characteristic for cilia of the tumor cells. The findings also emphasize that positive reaction of these markers will usually exclude a noncommon epithelial tumor, but will not distinguish between benign and malignant ovarian tumors. Report of cilia-bearing tumor cells in ascitic fluid are few; no one, to the best of our knowledge, has previously documented the immunocytochemistry of these tumor cells.

The value of the Cytobrush for obtaining cells from the uterine cervix.

A study was undertaken to evaluate the efficacy of the Cytobrush compared with the cotton-tipped applicator for obtaining cells from the uterine cervix. Four consecutive Papanicolaou smears were obtained in 1,000 patients during a single clinic visit. In every patient, a total of four smears on two slides were submitted: one slide contained endocervical material obtained with a cotton-tipped applicator and with the Cytobrush and the other slide contained ectocervical material obtained with both a cotton-tipped applicator and the Cytobrush. In endocervical smears, the Cytobrush technique produced numerous groups and clusters of cervical columnar cells in contrast to the use of the cotton swab. This was particularly true in older patients and pregnant women. In ectocervical smears, the detection of the squamous component was also higher in the Cytobrush smear than in cotton swab smears. However, the cell yield was not influenced by the cell sampling sequence in either endocervical or ectocervical specimens. It is the conclusion of this study that the utility of the Cytobrush technique for collecting specimens for endocervical and ectocervical smears is greater and provides a higher yield of cells than the cotton swab technique. Wider use of this simple procedure is encouraged.

Evaluation of cytocentrifuge apparatus with special reference to the cellular recovery rate.

Two types of commercially available cytocentrifuge apparatus (type A and type B apparatus) using disposable funnels were compared for percentage of cell recovery and degree of cell preservation. The cellularity of each cell suspension was determined using a Sysmex micro cell counter for blood analysis, and the cell recovery was obtained by counting cells in the total smeared area on the May-Grünwald Giemsa (MGG)-stained slide. Overall recovery rate by the type A apparatus was between 54.3% and 74.9% with a mean of 63.0%, whereas, the recovery rate for type B apparatus was between 30.6% and 51.8%, with a mean of 42.5%, indicating that the type A apparatus was significantly better. In the type A apparatus, a higher yield of all cells was obtained (69.7-74.9%) in the group of low cell counts (350 cells/0.5 ml), which was run for 10 minutes at 2,000 rpm. On the other hand, in the type B apparatus a higher yield of all cells was obtained (38.6-42.6%) in the group of low cell counts which was run for 10 minutes at 2,000 rpm. Cellular structure was better preserved on the slides in the type A apparatus. However, the percentage of ghost cells was somewhat higher in the type B apparatus. The cytocentrifugation of the type A apparatus consistently recovered a higher percentage of cells than with the type B apparatus. Using the type A apparatus, a high rate of cellular recovery, which is extremely important, such as for accurate morphological evaluation of cerebrospinal fluid, can be consistently obtained.

Brush cytology of herpes simplex virus infection in oral mucosa: use of the ThinPrep processor.

Evaluation was made of oral scraping specimen using Cytobrush from 11 patients with clinical suspicious lesions for herpes simplex virus (HSV) infection. In this study, oral brush cytology prepared using an automated smear apparatus (ThinPrep method) has been applied. The smears prepared by ThinPrep are equally well suited for screening purposes, producing highly cellular specimens with well-preserved cytoplasm and nuclei. The results obtained on the 11 patients, four cytologic specimens, showed pathognomonic viral cellular changes of HSV infection. Viral isolation studies were performed on 10 of the 11 patients and ten specimens tested positive for HSV by culture. Of the ten HSV positive isolates, six were positive by either cytology or immunocytochemical staining. Of the two smears showing HSV positive cells, both smears were also confirmed by in situ hybridization (ISH) with a biotinylated clone DNA probe. Using the ICC technique, detection of HSV antigen in oral scraping smears was of great value in the diagnosis of oral HSV infection, especially in cases of equivocal lesions. Oral cytology by means of ThinPrep method obviously deserves additional trials as an adjunct in the cytology of HSV-suspected lesions.