Imprint Slides Research Articles

Commentary on "Fusion of old and new: Employing touch imprint slides for next-generation sequencing in solid tumors".

Commentary on "Fusion of old and new: Employing touch imprint slides for next-generation sequencing in solid tumors".

Fusion of old and new: Employing touch imprint slides for next generation sequencing in solid tumors.

Cytomorphological evaluation of tissue touch imprints during rapid on-site evaluation or intraoperative pathology consultation has crucial value. However, literature on their utility for molecular testing is limited. In this study, we emphasize a further benefit of touch imprint slides and scrutinize our institutional experience on their use in molecular testing, specifically next generation sequencing (NGS). NGS-based reports (2019-2023) of Koç University Hospital were retrospectively analyzed and circumstances in which sequencing was conducted on touch imprint slides were retrieved (n = 18). Type/location of the biopsy, diagnosis, results, and quality metrics were recorded. Touch imprints were addressed when they harbored more neoplastic cells compared with permanent biopsies, when suboptimal fixation mitigated deoxyribonucleic acid/ribonucleic acid (DNA/RNA) yield in resections or when the sample was obtained from bone and required decalcification. Diagnoses were diverse, namely non-small-cell lung cancer, gastric adenocarcinoma, glial tumor, Ewing sarcoma, and carcinoma of unknown primary. The percentage of tumor cells on slides stretched between 15% and 70%. Molecular findings ranged from KRAS mutations to TRIM1::NTRK2 and EWSR::FLI1 fusions. For five cases, sequencing did not yield any alteration, one study was not completed because it did not yield high-quality RNA. Touch imprint slides provide a reliable alternative, especially when neoplastic cells are scarce in permanent biopsies or decalcification deters nucleic acid quality. Based on our experience, we suggest making touch imprints on a routine basis, especially for every bone biopsy. Once digitally scanned duplicates are made, original slides can be safely used for DNA-/RNA-based molecular studies.

Intraoperative pathologic diagnosis of central nervous system lymphomas: A comparison of frozen and permanent section diagnoses, and the significance of preoperative imaging

BackgroundCentral nervous system (CNS) lymphomas, either primary or secondary in origin, are rare malignant tumors affecting the brain, spinal cord, or leptomeninges. Diagnosis of CNS lymphomas is complicated by their diverse clinical presentations, radiological features, and histopathological characteristics. Although frozen section (FS) analysis is commonly employed for various CNS tumors, its role and accuracy in CNS lymphoma diagnosis are less explored. In this study, we conducted a comparative analysis to assess the impact of knowledge of preoperative imaging on enhancing the accuracy of FS diagnosis in CNS lymphomas. MethodsData collection involved a retrospective review of CNS lymphoma patients from January 2009 to August 2021. Patients who underwent intraoperative consultation were included, excluding those with prior cortisone treatment. The dataset incorporated patient demographics, classification as primary or secondary lymphoma, radiological preliminary diagnoses, FS diagnosis, and permanent section diagnosis. We employed various archived materials, including FSs, touch imprint slides, crush cytology slides, H&E-stained sections, and immunohistochemical stains, and re-evaluated all slides for diagnostic validation. ResultsOur study included 25 patients, of whom 60 % were female and had a mean age of 56.5 years. Preoperative radiology data were available for 80 % of cases, with preliminary diagnoses commonly including lymphoma and/or metastasis. Intraoperative consultation results indicated lymphoma in 18 (72 %) patients, with discordance observed in 28 % of cases when compared to permanent section diagnoses. Most permanent section diagnoses were diffuse large B-cell lymphomas (92 %), with the remainder being T-cell non-Hodgkin lymphoma (4 %) and follicular lymphoma (4 %). Intraoperative misdiagnoses were significantly associated with the absence of knowledge of preoperative imaging. ConclusionOur study demonstrates the reliability of FS diagnosis for CNS lymphomas during surgery, with a favorable complete concordance rate of 72 % when compared to permanent diagnoses. Importantly, lack of knowledge of preoperative imaging significantly impaired diagnostic accuracy in FS, emphasizing the need for close collaboration between pathologists and radiologists.

Usefulness of imprint cytology for early diagnosis in vertebral lesions

Abstract Background: Vertebral pathologies can be divided into degenerative, infective, or neoplastic. Infective and neoplastic lesions can progress rapidly and may lead to compression of the cord. Thus, rapid diagnosis for these lesions is important. Frozen section and fine-needle aspiration cytology techniques are less useful in hard bony tumors. Imprint cytology has been successfully used in the diagnosis of soft tissue tumors. The purpose of this study was to evaluate the role of imprint cytology in the early diagnosis of various spinal pathologies. Materials and Methods: Thirty-eight patients with suspected diagnosis of infection or tumor underwent transpedicular biopsy using Jamshidi needle. At least two imprint slides were made using core tissue obtained from biopsy. These slides were, thereafter, stained with the May-Grunwald–Giemsa method. The core biopsy sample was also sent for histopathological examination, after fixing it with 10% formalin. Paraffin-embedded blocks and slides were made for histopathology reporting, as per institution’s protocol. All imprint cytology slides were screened and reported by same pathologist. Results: The sensitivity of imprint cytology was 68.6%. Specificity, positive predictive value, negative predictive value, and overall accuracy of the imprint cytology were 100%, 100%, 21.4%, and 71%, respectively, when compared to histopathology. The mean duration of imprint cytology reporting was only 36 min while that for histopathological report was 15 days. Conclusion: Imprint cytology is a simple and rapid method for obtaining diagnosis in suspected vertebral lesions of infection or tumor. It is a reliable and cost-effective method in experienced hands. Although imprint cytology shows modest correlation in the ability to identify positive results, it forms a good diagnostic tool in confirming true negative cases with high diagnostic efficacy.

BRAF V600E mutation detection in hairy cell leukemia-utility of archival DNA from bone marrow aspirate/imprint smear and amplification refractory mutation system.

BRAF V600E is a disease defining mutation for hairy cell leukemia (HCL), which helps in its diagnosis and differentiation from morphologically similar splenic marginal zone lymphoma (SMZL) and HCL-variant (HCL-v). Forty eight cases:HCL(n = 34), SMZL(n = 11) and HCL-v(n = 3) were included. Of these, 32 were retrospective and 16 were prospective. DNA was extracted, in retrospective cases from cells obtained by smears from bone marrow aspirate/trephine imprint (BMA/BMTx) slides, and in prospective cases from peripheral blood (PB)/BMA samples. BRAF V600E mutation testing was done using ARMS-PCR. BRAF V600E mutation was positive in all HCL and negative in all SMZL and HCL-v cases. DNA extracted from BMA/BMTx slides gave results comparable to DNA extracted from PB/BMA samples. Median age of presentation for HCL (53years) and SMZL (56years) were quite similar, however, HCL-v (71years) cases presented at an older age. Statistically significant differences between the three groups were seen for total leucocyte, platelet, absolute lymphocyte and monocyte counts, bone marrow-infiltration pattern, reticulin fibrosis, and an expression of CD11c, CD25, CD103, CD123, and CD200. The use of BMA/BMTx smears for DNA extraction was found to be a useful alternative to DNA extraction from formalin-fixed paraffin-embedded biopsy sections. ARMS-PCR is an efficient and specific technique to detect BRAF V600E mutation in HCL patients.

Level of circulating steroid hormones in malaria and cutaneous leishmaniasis: a case control study.

Epidemiological and clinical studies have shown a great difference in the severity and prevalence of infectious diseases in men and women and various studies have shown that the key immunological factors are affected by sex-associated hormones. Considering the role of sex hormones in various infections, the current study aimed to determine the level of sex hormones in patients with cutaneous leishmaniasis (CL) and malaria and compare it with those of healthy controls. The survey was designed as a case-control study. Peripheral blood was collected from thirty male malaria patients, sixty patients (equal number of both sexes) with cutaneous leishmaniasis and ninety healthy subjects. Disease confirmations were done through microscopic examination of either peripheral blood smears, in case of malaria, or Giemsa-stained lesion imprint slides for CL. The level of testosterone, progesterone and estrogen were measured in malaria and CL patients along with healthy subjects, using an ELISA commercial kit. Age of participants was 18-35years (mean 25.39 ± 4.70) for CL patients and 14-41years (mean 27.63 ± 9.09) for malaria patients. Differences between the age of patients and the healthy subjects were insignificant. The level of testosterone in malaria patients (1.44 ± 0.12ng/mL) was lower than control group (1.46 ± 0.06, ng/mL) but the differences were not statistically significant (p > .05). The concentration of testosterone in CL patients (1.49 ± 0.03ng/mL) was higher than those of control group (1.46 ± 0.06ng/mL), and the difference was statistically significant (p =0.05). Although the concentration of estrogen and progesterone in CL patients were lower than controls, still the differences were not statistically significant (p > 0.05). Findings of the current study demonstrated a significant difference in the serum level of testosterone in CL patients in comparison with the healthy subjects whereas such difference was not seen in malaria patients.

Diagnostic performance of cytology for assessment of hepatic lipid content in dairy cattle

The objective of our study was to characterize the diagnostic performance of cytology for assessing hepatic lipid content (HLC) in dairy cows by comparing microscopic evaluation of lipid vacuolation in touch imprint slide preparations of liver biopsies with quantitative measurement of triglyceride concentration ([TG]; mg/mg of wet weight) in paired biopsy samples. Our study also sought to compare the diagnostic performance of liver cytology, plasma nonesterified fatty acid concentration ([NEFA]), and plasma β-hydroxybutyrate concentration ([BHB]) derived from a measurement performed on whole blood, for assessing HLC. Chemical extraction of TG from liver tissue remains the gold standard for quantifying HLC, largely because available blood tests, although useful for detecting some types of pathology, such as increased lipid mobilization, ketosis, or hepatocellular injury, are nonspecific as to etiology. Veterinary practitioners can sample bovine liver for cytological evaluation in a fast, minimally invasive, and inexpensive manner. Thus, if highly predictive of HLC, cytology would be a practical diagnostic tool for dairy veterinarians. In our study, liver biopsy samples from Holstein cows (219 samples from 105 cows: 52 from cows 2 to 20 d prepartum, 105 from cows 0 to 10 d in milk, 62 from cows 18 to 25 d in milk) were used to prepare cytology slides and to quantify [TG] using the Folch extraction method followed by the Hantzch condensation reaction and spectrophotometric measurement. An ordinal scale (0-4) based on amount of hepatocellular cytoplasm occupied by discrete clear vacuoles was used by 3 blinded, independent observers to rank HLC in Wright-Giemsa-stained slides. Interobserver agreement in cytology scoring was good. Corresponding plasma [NEFA] and [BHB] measurements were available for 187 and 195 of the 219 samples, respectively. Liver [TG] correlated more strongly with cytology score than with NEFA or BHB, and receiver operating characteristic curve analysis showed that cytology had better diagnostic performance than either NEFA or BHB for correctly categorizing [TG] at thresholds of 5, 10, and 15%. Hepatic lipidosis in high-producing dairy cows is of major clinical and economic importance, and this study demonstrates that cytology is an accurate means of assessing HLC. Additional work is indicated to evaluate the diagnostic utility of liver cytology.

“Intraoperative Touch Imprint Cytology of Sentinel Nodes”, Can it Serve as an Alternative to Frozen Section? – A Prospective Comparative Study of Touch Imprint Cytology and Frozen Section in the Intra Operative Assessment of Nodal Metastasis in Carcinoma Breast.

Sentinel lymph node biopsy (SLNB) is now known to offer an excellent alternative method to routine axillary lymph node dissection in staging of breast cancer. However, this minimally invasive technique requires facilities for frozen section (FS) and intraoperative analysis, which are sadly not available at most centers in India. At the same time Touch imprint cytology (TIC), is relatively less cumbersome technique for intraoperative assessment which may provide a fast and accurate method for intraoperative assessment of sentinel lymph nodes. Aim: This study was done to assess the accuracy and feasibility of using the TIC as an alternative to FS to assess metastasis in the sentinel node. Methods: Touch imprint cytological analysis of sentinel lymph nodes was compared prospectively with frozen section and the findings obtained on routine paraffin sections. Touch imprint slides from 32 consecutive women undergoing mastectomies (from Jan 2014 to May 2016) for invasive breast cancer (all 32 were T1-T3 and with clinically N0 nodal status) were prepared during operation from multiple sections of sentinel lymph nodes, stained with haematoxylin and eosin. Results: The sensitivity of TIC was 89.47% (95% CI, 68.49-98.81%), when compared with the final histopathological result as a gold standard method. The sensitivity of the FS was 94.74% (95% CI 73.97% to 99.87%), when compared to the HPE results. The specificity of the TIC and FS when compared to HPE is 100%. Conclusion: This study has lent credibility to the possible use of TIC as an assessment tool of the sentinel node in centers where facilities for frozen section are not available and hence expanding the population of potential beneficiaries who can be spared of morbidity associated with axillary dissection

Post-mortem hemoparasite detection in free-living Brazilian brown brocket deer (Mazama gouazoubira, Fischer 1814).

Tick-borne infections can result in serious health problems for wild ruminants, and some of these infectious agents can be considered zoonosis. The aim of the present study was the post-mortem detection of hemoparasites in free-living Mazama gouazoubira from Minas Gerais state, Brazil. The deer samples consisted of free-living M. gouazoubira (n = 9) individuals that died after capture. Necropsy examinations of the carcasses were performed to search for macroscopic alterations. Organ samples were collected for subsequent imprint slides, and nested PCR assays were performed to detect hemoparasite species. Imprint slide assays from four deer showed erythrocytes infected with Piroplasmida small trophozoites, and A. marginale corpuscles were observed in erythrocytes from two animals. A. marginale and trophozoite co-infections occurred in two deer. A nested PCR analysis of the organs showed that six of the nine samples were positive for Theileria sp., five were positive for A. phagocytophilum and three were positive for A. marginale, with co-infection occurring in four deer. The results of the present study demonstrate that post-mortem diagnostics using imprint slides and molecular assays are an effective method for detecting hemoparasites in organs.

Confounding Factors and Diagnostic Accuracy of Imprint Cytology

Background: Eradication of malignant tumors at the primary site with oncological safe margin is a critical requirement for obtaining better survival rate and less recurrence. Touch imprint cytology (TIC) has proven itself as a quick, simple, inexpensive, highly accurate and reliable intraoperative technique to assess surgical margins in squamous cell carcinomas of the oral cavity. However, it is still unclear how the mode of excision, i.e. by scalpel (SC) and electrocautery (EC), or the method of staining, i.e. Papanicolaou (PAP) and cytohaem, affect the diagnostic accuracy of TIC. Objective: To study the influence of confounding factors like mode of excision (EC/SC) and staining (PAP/cytohaem) on the diagnostic accuracy of intraoperative TIC technique for assessing surgical margins in oral squamous cell carcinoma in comparison to paraffin-embedded HE-stained sections. Materials and Methods: Thirty patients underwent surgical treatment for primary oral squamous cell carcinoma. Three hundred and forty-eight touch imprint slides were prepared from 174 margins of 30 resected tumor specimens. Two adjacent tissues from the margin to be evaluated were imprinted to observe differences between surfaces excised by EC and SC. The set of imprint from each margin tissue was stained with PAP and cytohaem. The TIC results of 180 EC-excised margins and 168 SC-excised margins were compared. Results of 174 imprints stained with RAPID-PAP were compared to their counterpart comprising of 174 cytohaem-stained imprints. The slides were diagnosed as positive, negative or suspicious for tumor. Finally, TIC results were checked against their respective histopathological sections. Results: No statistically significant difference was found between the results of imprints from EC/SC-excised margins (Z = 0.44, p = 0.70) or the imprints stained with PAP/cytohaem (Z = 0.44, p = 0.70). Conclusion: Confounding factors like mode of excision and staining procedure do not significantly influence the results of imprint cytology.

Use of touch imprint cytology as a simple method to enrich tumor cells for molecular analysis

A relative excess of nonneoplastic cells in frozen carcinoma samples is often a cause of false-negative results in molecular assays. Given the greater cohesiveness of epithelial tumor cells compared with nonneoplastic epithelium and mesenchymal stroma, the authors hypothesized that tumor procurement by touch imprinting would provide a simple, cost-effective method of obtaining enriched neoplastic cells compared with frozen whole-tumor samples. Eleven adenocarcinomas with known KRAS gene mutations were tested. Two sets of 8 touch imprint (TP) slides and 1 frozen whole-tumor sample (FS), both with a corresponding hematoxylin and eosin-stained slide, were obtained from each tumor. DNA from unstained TP and FS samples was tested for KRAS exon 2 mutations by Sanger sequencing. The percentage of carcinoma cells was determined by light microscopy of hematoxylin and eosin-stained slides. The fold increase in the mutant-enriched DNA in TP versus FS samples was determined by calculating the height ratio between the mutant and wild-type peaks on the sequencing electropherogram. Using light microscopy, TP demonstrated a 1.1-fold to 3.5-fold (mean, 1.8-fold) enrichment in neoplastic cells compared with the FS. The mutant-to-wild-type peak height ratio was 1.4-fold to 7.1-fold (mean, 3.1-fold) higher in TP compared with the corresponding FS samples. The average amount of extracted DNA ranged from 145 ng to 7.9 μg per TP slide. The procurement of carcinoma samples by TP is rapid, simple, and inexpensive; consistently provides a tumor-enriched sample; is an excellent source of high-quality tumor DNA; and could compensate for the relatively low sensitivity of direct sequencing. Cancer (Cancer Cytopathol) 2013;121:354-360. © 2013 American Cancer Society.

A canine hemangiopericytoma case: Fine needle aspiration (FNA) cytology and histopathological findings

Veterinary cytology is gaining importance in diagnostics of skin and subcutaneous masses of dogs. A case of canine-hemangiopericytoma was described by cytopathological and histopathological findings on Fine needle aspiration (FNA) and excisional biopsy. A 13-year-old female terrier dog had a subcutaneous lesion on right leg. FNA was performed by the cytopathologist using 27-gauge needle and 10 ml. syringe attached to a syringe-holder. FNA and imprint slides from FNA aspirates and stained with May-Grunwald Giemsa,Papanicalau, Hematoxylin Eosin were prepared. Cyto-diagnosis was encountered with spindle cells. FNA can often provide information for a good diagnostic and management plan, especially for discrete, non-ulcerated lesions, allowing veterinarians to make more informed therapeutic choices

Intraoperative Imprint Evaluation of Surgical Margins in Oral Squamous Cell Carcinoma

Background: Oral squamous cell carcinoma (OSCC) is a multifactorial malignant epithelial neoplasm. Tumor clearance at the surgical margins in OSCC is a critical factor which strongly influences both the local recurrence and overall survival rate. A quick and accurate intraoperative diagnostic procedure can make a significant difference to the survival rate. Touch imprint cytology (TIC) has commonly been used in the past for the diagnosis of tumors and the detection of sentinel lymph node metastasis. It is routinely applied for intraoperative margin analysis of breast lumpectomy specimens, where it has been proven to have a good success rate. The effectiveness and reliability of TIC in the intraoperative margin evaluation of OSCC, however, has not been established so far. Objective: To investigate the diagnostic accuracy of the intraoperative TIC technique for assessing surgical margins in OSCC in comparison to paraffin-embedded hematoxylin and eosin-stained sections. Materials and Methods: Thirty previously untreated, biopsy-proven OSCC patients undergoing surgical treatment between December 2008 and September 2010 were included. Patients diagnosed with histological variants of squamous cell carcinoma (SCC) or recurrent tumor were excluded. Three hundred forty-eight touch imprint slides were prepared from 174 margins of 30 resected tumor specimens. Imprints from tumor proper were taken as positive controls. The slides were reported as positive, negative or suspicious for tumor. After all cytological interpretation was completed, the results were compared with the corresponding histological diagnosis. Results: This pioneer study showed that TIC has an overall accuracy of 83%, sensitivity of 91.1%, specificity of 74.4%, positive predictive value of 79.2% and a negative predictive value of 88.6%. The false-positive margins evaluated by TIC were 43 (12.3%) and the false-negative ones were 16 (4.6%). Conclusion: TIC is a quick, simple, inexpensive, highly accurate and reliable intraoperative technique to assess surgical margins in SCC of the oral cavity.

Vastagtű-biopsziás hengerből készített imprint citológia növeli a vékonytű-aspirációs citológia szenzitivitását emlőrákos betegekben

Fine needle aspiration cytology (FNAC) can have high inadequate results. The main objective of this study was to validate the sensitivity of imprint cytology and compare it to that of FNAC across all levels of staff experience. Our other objective was to find out whether handling of a core biopsy to obtain an imprint slide affected its morphology so as to make histopathological reporting from that tissue difficult. This we thought could be of significance while trying to diagnose smaller cancers where just one core could contain tumour. Patients (n = 56) with a suspicious breast lump had fine needle aspiration (FNA) and core biopsy. The core biopsy used to prepare the two imprint cytology slides was sent separately for histopathology reporting. Simultaneous imprint cytology from a core biopsy increased the sensitivity of cytology results by 12% compared to that of FNAC alone. In this series we found imprint cytology to have a sensitivity of 84%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value of 82.7%. The histopathologists did not report any distortion in tissue morphology so as to affect histological reporting of the core used. Since most patients will have a core biopsy to confirm invasive cancer and the technique of imprint cytology is easily performed, perhaps it should be more widely used especially in units where the sensitivity of FNA is low.

Use of imprint cytology for assessment of surgical margins in lumpectomy specimens of breast cancer patients

In the past several years, breast-conservation therapy has provided an alternative to mastectomy. In order to reduce the subsequent local tumor recurrence, it is critical that all the measures are in place to find the residual foci of occult microscopic tumor at the time of the initial lumpectomy procedure. An accepted method to evaluate the lumpectomy margins for presence of residual tumor is the use of imprint cytology (also called touch-prep), which is assessment of the presence or absence of the tumor cells by cytological preparation. This is a rapid, cost effective, and easy to use procedure with added advantage of saving tissue for permanent sectioning and rendering a definitive diagnosis. In this report, we present our experience using intraoperative imprint cytology for evaluation of the status of lumpectomy specimens in breast cancer patients. The objective of this study was to evaluate the diagnostic accuracy of intraoperative imprint cytology for assessment of surgical resection margins in lumpectomy margins of patients with breast carcinoma. This is a retrospective study of 100 cases of breast lumpectomy specimens, which had undergone intraoperative imprint cytology. The cases were retrieved from the archived files of the University of Florida, Department of Pathology at Shands Jacksonville. The results of intraoperative imprint cytology were compared with the histological findings of the corresponding permanent sections of the same cases as the gold standard. Overall, we reviewed 510 cytology imprint slides, which were obtained from 100 lumpectomy specimens. Among these cases, 37 slides from 22 cases were reported positive and the remaining were negative. Only eight slides from six cases of lumpectomy showed discrepancy between the result of intraoperative imprint cytology and the permanent sections of the same cases. In our study, intraoperative imprint cytology showed a sensitivity of 97%, specificity of 99%, with positive predictive value of 84%, and negative predictive value of 99%. This study demonstrates that intraoperative imprint cytology can be used as a reliable diagnostic procedure for the evaluation of the status of lumpectomy margins in breast cancer patients.

Leishmania identification by PCR of Giemsa-stained lesion imprint slides stored for up to 36 years

This study examined the ability of PCR to amplify Leishmania DNA, stored on Giemsa-stained slides, from American cutaneous leishmaniasis (ACL) patients. In total, 475 slides stored for up to 36 years were obtained from an outpatient clinic in a Brazilian ACL-endemic region, and Leishmania DNA was amplified from 395 (83.2%) of the DNA samples using primers specific for the minicircle kinetoplast DNA. Restriction fragment length polymorphism analysis of these amplicons demonstrated that Leishmania (Viannia) braziliensis was the only species present in these samples. The results demonstrated that archived Giemsa-stained slides can provide a Leishmania DNA source for performing clinical and epidemiological studies of leishmaniasis.

Differential resistance to koi herpes virus (KHV)/carp interstitial nephritis and gill necrosis virus (CNGV) among common carp ( Cyprinus carpio L.) strains and crossbreds

Massive mortalities of edible and ornamental (koi) carp ( Cyprinus carpio L.) have been encountered in several fish farms worldwide, including Israel. The disease-causing virus has been identified as koi herpes virus (KHV). Based on its pathogenic effect in fish, it has also been termed carp interstitial nephritis and gill necrosis virus (CNGV). The first objective of this work was to select virus-resistant fish by crossbreeding sensitive domesticated strains with resistant wild strains of carp. Frozen wild carp [ Cyprinus carpio haematopterus; Sassan] sperm was brought from the Czech Republic to achieve heterosis by producing crossbreds. We compared growth rate, food consumption, survival and behavior among strains and crossbreds. Two strains of domesticated carp—Našice (N) and Dor-70 (D) and the wild Sassan (S) were used. A large quantity of Našice eggs were fertilized with semen of Našice, Dor-70 or Sassan to produce the N×N, N×D and N×S crossbred groups, while a large quantity of Dor-70 eggs were fertilized with semen of Dor-70 or Sassan to produce the D×D and D×S groups. Resistance to the viral disease was evaluated by exposure to the virus in ponds and in the laboratory. The laboratory-exposure test was carried out at a water temperature of 22 to 23 °C and the fish were exposed to the disease by cohabitation. The D×S crossbred was the most resistant (60.7% survival) and the N×N crossbred most sensitive (8.0% survival); the other fish strains did not differ significantly from one another (N×S—33.7%, D×D—27.0%, and D×N—17.7%). Similar tests, performed in ponds, were generally in good agreement with those from the laboratory, except for the N×S crossbred, which showed good resistance in the ponds but only intermediate resistance in the laboratory. In the grow-out phase, the Sassan crossbreds exhibited equal (N×S) or better (D×S) growth rates as compared to the commercial crossbred (D×N) and the pure strains (D×D, N×N). We used indirect immunofluorescence microscopy to evaluate the increasing amounts of virus in the sick fish. Kidneys were removed from naïve and sick fish and used for the preparation of touch imprint slides. The amount of viral protein produced by the infected kidney cells was high, enabling detection of the infected cells as early as 3 days post-infection. Thus, a simple imprinting immunofluorescent assay may be appropriate for CNGV diagnosis.

CD4+CD56+CD68+hematopoietic tumor of probable plasmacytoid monocyte derivation with weak expression of cytoplasmic CD3.

Hematopoietic neoplasm coexpressing CD4 and CD56 includes a subset of acute myeloid leukemia with myelomonocytic differentiation, plasmacytoid monocyte tumor, and other immature hematopoietic neoplasms of undefined origin. Herein, we report a CD4+CD56+CD68+ hematopoietic tumor that was thought to be a tumor of plasmacytoid monocytes. This case is unique in the absence of accompanying myelomonocytic leukemia and the faint expression of cCD3 on the tumor cells. The patient was a 22-yr old man presented with multiple lymphadenopathy and an involvement of the bone marrow. Tumor cells were large and monomorphic with an angulated eosinophilic cytoplasm of moderate amount. Nuclei of most tumor cells were eccentric and round with one or two prominent nucleoli. Rough endoplasmic reticulum was prominent in electron microscopic examination. Tumor cells expressed CD4, CD7, CD10, CD45RB, CD56, CD68, and HLA-DR and were negative for CD1a, CD2, sCD3, CD5, CD13, CD14, CD20, CD33, CD34, CD43, CD45RA, TIA-1, S-100, and TdT. cCD3 was not detected in the immunostaining using paraffin tissue, but was faintly expressed in flow cytometry and immunostaining using a touch imprint slide. T-cell receptor gene rearrangement analysis and EBV in situ hybridization showed negative results. Cytochemically, myeloperoxidase, Sudan black B, and alpha naphthyl butyrate esterase were all negative.

CORE IMPRINT CYTOLOGY: A USEFUL METHOD FOR SAME‐DAY REPORTING

INTRODUCTION: Core biopsy of the breast has become the method of choice for tissue diagnosis of screen detected microcalcifications and some mass lesions in many breast assessment centres. Biopsy results are not available until the following day. Imprint cytology of fresh breast core samples allows same‐day reporting and patient counselling.AIM: To determine the accuracy of core imprint cytology when compared with core biopsy diagnosis when used in a breast assessment centre setting.METHODS: Core imprints (CI) were prepared and reported on all fresh core biopsies (CB) performed at the Sir Charles Gairdner Hospital Breast Centre from May to December 2000. Fresh core samples were placed on a glass microscope slide. Core radiographs were taken for microcalcification lesions (MC). A laboratory technician gently and quickly rolled the cores on the slide with fine forceps. The cores were fixed in formalin, processed and reported next day. The imprint slide was air dried and stained with DiffQuik. CI were reported using four categories: Insufficient, Benign, Indeterminate and Malignant. Counselling and planning for management were possible on the same day in women with malignant diagnoses. Clinicians were advised not to discuss negative or indeterminate CI results with women and to defer to the final CB report.RESULTS: Cores were performed on 381 lesions. There were 83 carcinomas (38 in MC and 45 in masses) and 56 were called malignant on CI (absolute sensitivity 67.5%; 78.9% for MC and 57.8% for masses). 3 malignancies on CB were negative on CI giving a false negative rate of 3.6%. There were no false positive diagnoses. The predictive value of a benign diagnosis was 95.3%. There were no adverse effects in the histology of CB.CONCLUSION: CI was an accurate method of providing an immediate diagnosis of malignancy in two thirds of malignancies confirmed on CB.

A comparison of touch imprint cytology and Mohs frozen-section histology in the evaluation of Mohs micrographic surgical margins

Background: Touch imprint cytology (TIC) is commonly used in the diagnosis of tumors and has been applied to margin analysis of breast lumpectomy specimens with good success.Objective: Our purpose was to determine the diagnostic adequacy of TIC for identifying positive and negative Mohs surgical margins for basal cell carcinoma (BCC) excisions, compared with the “gold” standard, Mohs tangential sectioning.Methods: Fifty-eight patients undergoing 69 Mohs micrographic surgical procedures for biopsy-proven BCC were included in this study between October 1998 and January 1999. Patients were excluded if the neoplasms were of another histologic type, including BCCs with squamous features. One hundred sixty-six touch imprint slides were prepared from 166 fragments of skin tissue excised during MMS. Touch imprint slides were evaluated blindly and independently by two pathologists, one of whom was also a cytopathologist. The slides were diagnosed as positive for tumor, negative for tumor, or, rarely, atypical but suspect for tumor. Discrepancies between the pathologists' interpretations were re-evaluated with the use of a two-headed microscope and a consensus was reached. After all cytologic interpretation was completed, the results were compared with the histologic diagnosis rendered for each fragment of tissue by the Mohs surgeon.Results: The prevalence of a positive margin by histologic confirmation was 55% overall, 60% for recurrent or sclerosing lesions, and 51% for nonsclerosing or recurrent lesions. The overall accuracy of this technique in identifying true positive and true negative margins was 71%. The sensitivity of TIC for identifying a positive margin was approximately 50% for all BCC types. The specificity was approximately 90% for all BCC types.Conclusion: TIC is inadequate for identifying positive margins compared with the “gold” standard, MMS. (J Am Acad Dermatol 2001;44:660-4.)