Artifacts and main lesions in fetal ocular histology.

Histological artifacts in fetal eyes can involve different tissues and can be related to mechanical or autolytic lesions, fixation, the cause of death or the cutting technique. Knowing the causes, effects and appearance of artifacts allow the minimization of the risk of avoidable artifacts and help distinguish them from "true" pathological lesions. We describe these different types of artifacts and specifically analyze their involvement in different tissue structures of the eye. We compare them with primary fetal ocular lesions. Given the wide variety of artifacts, the identification of lesions in fetal eyes must be done with caution, since differentiating true lesions from artifacts requires some experience and relies on both macroscopic examination and microscopic analyses, with ideally comparisons with references' images of normal tissues of the same gestational age.

Ultrasound-guided fine-needle capillary cytology of parotid gland masses coupled with a rapid-on-site evaluation improves results.

OBJECTIVE OF THE STUDY: To test whether a direct-on-site microscopic examination of fresh, unstained puncture slides by the radiologist decreases the rate of false-negative cases on ultrasound-guided fine-needle cytology of parotid gland masses. PATIENTS: Thirty parotid gland masses from 28 patients were punctured under ultrasound guidance. The same group was used as its control group. METHODS: After one or two passes, the material was spread on slides and air-dried (control group, without microscopic examination). For the study group, it was thus analyzed unstained under the microscope. A sample was considered adequate if at least six clusters of parotid cells were found per slide on at least two slides. For the study group, new punctures were obtained and slides prepared until this condition was fulfilled. RESULTS: Of the 30 evaluated masses, 100% benefited from a cytological diagnosis after microscopy. Twenty-four were adequate in the control group, while 30 were adequate in the study group. The maximum number of punctures to obtain an adequate sample was six. On-site direct microscopy significantly increased the number of adequate specimens by 20% (P=0.03, CI [1.63-20%]). CONCLUSION: Direct and systematic examination of slides by a radiologist avoided the risk of false-negative results caused by having insufficient sample material.

Use of fluorescent in-situ hybridisation in salivary gland cytology: A powerful diagnostic tool.

OBJECTIVE: Salivary gland cytology is challenging because it includes a diversity of lesions and a wide spectra of tumours. Recently, it has been reported that many types of salivary gland tumours have specific molecular diagnostic signatures that could be identified by fluorescent in-situ hybridisation (FISH). The aim of the present study was to demonstrate the feasibility and efficiency of FISH on routine cytological salivary gland smears. METHODS: FISH was conducted on 37 cytological salivary gland smears from 34 patients. According to the cytological diagnosis suspected, MECT1/MAML2 gene fusion and rearrangements of PLAG1, MYB, or ETV6 were analysed. The presence and percentages of cells that had gene rearrangements were evaluated. Results were compared with the histological surgical samples, available from 26 patients. RESULTS: The PLAG1 rearrangement was observed in 12/20 (60%) cases of pleomorphic adenoma. MECT1/MAML2 gene fusion was observed in 1:2 mucoepidermoid carcinomas but was not observed in five other tumours (two pleomorphic adenomas, one Warthin's tumour, one mammary analogue secretory carcinoma [MASC] and one cystic tumour). MYB rearrangement was observed in 4/4 adenoid cystic carcinomas. ETV6-gene splitting identified one MASC. CONCLUSION: Overall, FISH had a specificity of 100% and a sensitivity of 66.7%. When FISH and cytological analyses were combined, the overall sensitivity was increased to 93.3%. It can thus be concluded that when the FISH analysis is positive, the extent of surgery could be determined with confidence pre-operatively without needing a diagnosis from a frozen section.

Immunocytochemical staining for p53 and Ki-67 helps to characterise urothelial cells in urine cytology.

OBJECTIVE: The presence of atypical cells in urine cytology is unsatisfactory for both cytologists and clinicians. The objective of this study was to test whether p53 and Ki-67 immunostaining could improve urothelial carcinoma (UC) detection on urinary cytology. METHODS: A total of 196 urine samples were analysed, 142 from the bladder, 41 from the upper tract and 13 from ileal bladder replacement. Cytology results were expressed as normal (N) (n = 81), atypia cannot exclude low-grade UC (ALG) (n = 25), suspicious for high-grade UC (SHG) (n = 39) and high-grade UC (HG) (n = 51). Actual diagnoses were confirmed by histopathological analysis, cystoscopic examination or follow-up for at least 1 year. Immunocytochemistry performed on CytoSpin™ slides allowed the determination of the percentage of positive cells with p53 and Ki-67. RESULTS: The median percentage values [first to third quartile] of p53 and Ki-67 were 0 [0-5] and 0 [0-1] for N cytology, 5 [0-40] and 2 [1-10] for ALG, 10 [0-30] and 6 [3-25] for SHG, and 30 [10-80] and 20 [10-30] for HG, respectively. Statistically higher values were observed for both tests (P < 0.001) in positive cytologies (ALG, SHG and HG). The optimal cut-offs were 5% for p53 and 3% for Ki-67. The sensitivity and specificity for the detection of all UC were 86.4% and 76.7% for cytology alone, 81.3% and 93.2% for cytology and p53, 75.7% and 88% for cytology and Ki-67, and 68.9% and 97.5% for cytology, p53 and Ki-67, respectively. CONCLUSION: Using p53 and/or Ki-67 in addition to cytology increases the specificity without penalising the sensitivity.

Guidelines for May-Grünwald-Giemsa staining in haematology and non-gynaecological cytopathology: recommendations of the French Society of Clinical Cytology (SFCC) and of the French Association for Quality Assurance in Anatomic and Cytologic Pathology (AFAQAP).

OBJECTIVE: Since the guidelines of the International Committee for Standardisation in Haematology (ICSH) in 1984 and those of the European Committee for External Quality Assessment Programmes in Laboratory Medicine (EQALM) in 2004, no leading organisation has published technical recommendations for the preparation of air-dried cytological specimens using May-Grünwald-Giemsa (MGG) staining. DATA SOURCES: Literature data were retrieved using reference books, baseline-published studies, articles extracted from PubMed/Medline and Google Scholar, and online-available industry datasheets. RATIONALE: The present review addresses all pre-analytical issues concerning the use of Romanowsky's stains (including MGG) in haematology and non-gynaecological cytopathology. It aims at serving as actualised, best practice recommendations for the proper handling of air-dried cytological specimens. It, therefore, appears complementary to the staining criteria of the non-gynaecological diagnostic cytology handbook edited by the United Kingdom National External Quality Assessment Service (UK-NEQAS) in February 2015.

Improvement of the cytological diagnostic accuracy of follicular thyroid lesions by the use of the Ki-67 proliferative index in addition to cytokeratin-19 and HBME-1 immunomarkers: a study of 61 cases of liquid-based FNA cytology with histological controls.

OBJECTIVES: To evaluate HBME-1, cytokeratin-19 (CK-19) and Ki-67 immunomarkers in order to increase the diagnostic accuracy of preoperative thyroid fine needle aspiration (FNA) cytology. METHODS: Immunocytochemistry against HBME-1, CK-19 and Ki-67 was performed on 123 thyroid FNAs processed by liquid-based cytology (LBC). Statistical analysis was carried out on 61 cases with histological control and sufficient material for one or more of the three markers. The Bethesda System was used for cytological diagnosis. RESULTS: Taking into account all the cytological categories, with a cut-off of 30% of positive cells, HBME-1 (n = 47) and CK-19 (n = 53) showed a sensitivity for malignancy of 66.7% (95% confidence interval, 53.2-80.1) and 90.5% (82.6-98.4) and a specificity of 90.6% (82.3-99) and 75% (63.3-86.7), respectively. For Ki-67 (n = 54) with a cut-off of 1% of positive cells, the sensitivity was 85.0% (75.5-94.5) and the specificity 70.6% (58.4-82.7). In the follicular neoplasm/suspicious for follicular neoplasm (FN/SFN) category (n = 37), which was the focus of the study, papillary thyroid carcinomas (PTCs) were less numerous (four cases, three of which were the follicular variant), the positivity of the three immunomarkers combined showed an overall accuracy of 91% (21/23). The mean percentage of Ki-67-positive cells was increased in malignant lesions, with the exception of follicular variant PTCs: 16% ± 15.6% in two follicular carcinomas, 4.8% ± 3.2% in 13 classical PTCs, 1% ± 1.2% in five follicular variant PTCs and 0.5% ± 1.9% in 34 non-malignant lesions. CONCLUSIONS: Immunocytochemistry using HBME-1, CK-19 and the Ki-67 proliferative index increased the diagnostic accuracy of FNA in the FN/SFN category of the Bethesda System, which may help to distinguish lesions in this category with a low or high risk of malignancy. Thus, clinical management would be improved.

Dermatofibrosarcoma protuberans: margins reduction using slow-Mohs micrographic surgery. Experience with 35 patients.

INTRODUCTION: Dermatofibrosarcoma protuberans (DFSP) is a potentially malignant dermal mesenchymal tumour with a high risk of local recurrence. DFSP presents a sprawling appearance whose complete excision requires important margins. DFSP was initially resected with a 5cm excision margins, and more recently 3cm then 2cm margins were recommended. Mohs micrographic surgery (MMS) helps reduce these margins thanks to a 3-dimensional excision around the tumour, which is analysed in its entirety. We used the modified MMS called slow-MMS and tried every time it was possible to perform direct closure. METHODS: Thirty-five patients presenting a DFSP between 2004 and 2013 within the Plastic Surgery unit at Claudius Regaud Institute were included in this retrospective study. The patients were treated with slow-MMS using paraffin-embedded sections. RESULTS: One surgery was necessary for 72% of patients. For 17%, we had to perform a second surgery, and for 11% a third one. Our median clinical excision margins was 17mm (range 9.0:30.0). After a median follow-up of 46 months (range 35.2:60.2), we didn't observe any recurrence. Only one case required a local flap; for the others, the loss of substance was resolved with a direct closure. CONCLUSION: Slow-MMS enabled a local control of the margins without recurrence at 46 months in our series. Besides, it helps performing smaller margins than wide excision and thus preserving the tissues. In our opinion, this is the treatment of choice regarding DFSP for which tissue sparing is essential. It seems particularly appropriate near functional areas or on the face.

Cytopathological analysis of salivary gland cancer: REFCOR recommendations by the formal consensus method.

OBJECTIVE: To determine the indications for fine-needle cytology and the modalities of frozen section pathological analysis in the management of salivary gland cancer. MATERIAL AND METHODS: The French Network of Rare Head and Neck Tumors (REFCOR) formed a steering group who drafted a narrative review of the literature published on Medline and proposed recommendations. The level of adherence to the recommendations was then assessed by a rating group according to the formal consensus method. RESULTS: Fine-needle cytology is recommended as part of the diagnostic work-up for a major salivary gland tumor suspicious for malignancy. Fine-needle cytology should be performed after MRI to avoid artifacts. Frozen section analysis is recommended to confirm the malignant nature of the tumor, to adapt the extent of resection and to indicate neck dissection. Whenever possible, the entire tumor and adjacent salivary or periglandular tissue should be sent for frozen section analysis. CONCLUSION: Fine-needle cytology and frozen section analysis play an essential role in the management of salivary gland cancers.

Real-time and non-invasive acute lung rejection diagnosis using confocal LASER Endomicroscopy in lung transplant recipients: Results from the CELTICS study.

BACKGROUND AND OBJECTIVE: Traditionally, the diagnosis of acute rejection (AR) relies on invasive transbronchial biopsies (TBBs) to obtain histopathological samples. We aimed to evaluate the diagnostic yield of probe-based confocal laser endomicroscopy (pCLE) as a complementary and non-invasive tool for ACR screening, comparing its results with those obtained from TBBs. METHODS: Between January 2015 and April 2022, we conducted a retrospective study of all lung transplant recipients aged over 18 years at Toulouse University Hospital (France). All patients who underwent bronchoscopies with both TBBs and pCLE imaging were included. Two experienced interpreters (TV and MS) reviewed the pCLE images independently, blinded to all clinical information and pathology results. RESULTS: From 120 procedures in 85 patients, 34 abnormal histological samples were identified. Probe-based confocal laser endomicroscopy revealed significant associations between both alveolar (ALC) and perivascular (PVC) cellularities and abnormal histological samples (p<0.0001 and 0.003 respectively). Alveolar cellularity demonstrated a sensitivity (Se) of 85.3 %, specificity (Spe) of 43 %, positive predictive value (PPV) of 37.2 % and negative predictive value (NPV) of 88.1 %. For PVC, Se was 70.6 %, Spe 80.2 %, PPV 58.5 % and NPV 87.3 %. Intra-interpreter correlation (TV) was 88.3 % for the number of vessels (+/-1), 98.3 % for ALC and 90 % for PVC. Inter-interpreter correlation (TV and MS) was 80 % for vessels (+/-1), 97.5 % for ALC and 83.3 % for PVC. CONCLUSION: Our study demonstrates the feasibility of incorporating pCLE into clinical practice, demonstrating good diagnostic yield and reproducible outcomes in the screening of AR in lung transplant recipients.

Investigation of the significance of Oil Red O-positive macrophage excess in bronchoalveolar lavage fluid during HIV infection.

OBJECTIVE: To assess the significance of increased levels of Oil Red O-positive macrophages (ORO-PM) in bronchoalveolar lavage fluids (BALFs) from HIV-positive patients. METHODS: Cytological data for seventy BALF samples from 66 consecutive HIV-infected patients were analysed according to antiretroviral therapy regimen, presence of Pneumocystis jiroveci infection, blood CD4(+) T cell count, HIV-1 viral load and plasma lipid levels. Non-parametric tests were used to compare the values between groups. RESULTS: The percentages of ORO-PM were high in this group: 40% [6-80] (median [interquartile range]). They were positively correlated with the BALF total cell count, 21% [5-48.5] for <300 cells/mm(3) and 60% [26.5-80] for >300 cells/mm(3) (P<0.01) but inversely correlated with the percentage of BALF lymphocytes, 50% [20-80] for <15% lymphocytes and 11.5% [2-47] for ≥15% lymphocytes (P<0.01). Antiretroviral therapy with or without protease inhibitors, plasma lipid levels, HIV-1 viral load, blood CD4(+) T cell count or presence of a Pneumocystis jiroveci infection were not correlated with the ORO-PM status. CONCLUSION: Significantly increased numbers of ORO-PM were correlated with high total cell counts and low lymphocyte counts in BALF, irrespective of disease activity or treatment. Extended work on a larger series of patients needs to be conducted.

[Interest of a systematic second-look at 48-72 hours in high-pressure injections of paint: an animal study]. / Intérêt d'une revisite systématique à 48-72 heures dans les injections à haute pression de peinture : une étude animale.

STUDY PURPOSE: High-pressure injections (HPI) in the hand are rare and their prognosis is often tragic, some authors report up to 48% of amputations. Poorly standardized, their management is often too late and inadequate. The HPI of paint results in inoculation of toxic particles in depth that are particularly difficult to remove surgically. The persistence of this foreign material maintains local inflammation and increases the risk of infection. We wanted to simulate in rats, the physiopathology of HPI with paint. The aim of this study is to authenticate an advantage to achieve consistently a second surgical debridement at 48-72 hours in HPI with paint. MATERIALS AND METHODS: Six rats were injected with 200 bars of white glycerophtalic paint in the leg. At six hours and then every 24 hours for four days, we did a debridement and a surgical washing, then at the end of intervention a tissue sample was analyzed by histology. RESULTS: Despite surgical debridement and thorough washing, at each step we found the persistence of painting in the intermuscular septa by light microscopy. From 24 hours, a necroinflammatory process is set up with an influx of neutrophils, organizing themselves into micro-abscesses. It will be very deleterious, initially by encouraging risk of sepsis, and then facilitating the formation of extensive fibrosis, which is the cause of functional impairment. CONCLUSION: We established the kinetics of tissue under HPI with paint using an animal model. It appears that single emergency surgery is insufficient to manage this "septic compartment syndrome". Therefore, we recommend a systematic surgical "second look" at 48-72 hours to complete the debridement of residual inoculum and necrotic tissue without wound closure.

[Histological techniques in oncodermatology]. / L'apport des techniques anatomopathologiques en oncodermatologie.

The skin oncology or "oncodermatology" requires a surgical treatment in most cases. For some surgeons, the oncodermatology takes a very important part of their practice. In the course of diagnostic and therapeutic of skin lesions, the pathologist plays now an essential role. He will guide our surgery. The techniques used by this specialist are numerous. Therefore, the objective of this paper is to review the different histological methods used to improve our management of skin tumors.

Guidelines of the French Society of Otorhinolaryngology-Head and Neck Surgery (SFORL), part I: Primary treatment of pleomorphic adenoma.

INTRODUCTION: The authors present the guidelines of the French Society of Otorhinolaryngology-Head and Neck Surgery (SFORL) for the diagnosis and treatment of pleomorphic adenoma (PA) of the salivary glands. METHOD: A review of the literature was performed by a multidisciplinary task force. Guidelines were drafted based on the articles retrieved and the workgroup members' individual experience. Guidelines were graded A, B, C or expert opinion by decreasing level of evidence. RESULTS: In clinically suspected salivary gland PA, MRI should be performed, including head and neck lymph node levels. Fine needle aspiration cytology is particularly recommended for tumours difficult to characterise by MRI. Frozen section biopsy should be performed to confirm diagnosis and adapt the surgical procedure in case of intraoperative findings of malignancy. Complete resection of the parotid PA should be performed en bloc, including margins, when feasible according to tumour location, while respecting the facial nerve. Enucleation (resection only in contact with the tumour) is not recommended. For the accessory salivary and submandibular glands, complete en bloc resection should be performed.

Guidelines of the French Society of Otorhinolaryngology-Head and Neck Surgery (SFORL), part II: Management of recurrent pleomorphic adenoma of the parotid gland.

INTRODUCTION: The authors present the guidelines of the French Society of Otorhinolaryngology-Head and Neck Surgery (SFORL) for the management of recurrent pleomorphic adenoma (RPA) of the parotid gland. METHOD: A review of the literature was performed by a multidisciplinary task force. Guidelines were drafted, based on the articles retrieved and the work group members' individual experience. There were then read and re-edited by an independent reading group. The proposed recommendations were graded A, B or C on decreasing levels of evidence. RESULTS: Complete resection under neuromonitoring is recommended in case of RPA. The risks of progression and malignant transformation, which are higher the younger the patient, have to be taken into consideration. The risk of functional sequelae must be explained to the patient. MRI is recommended ahead of any surgery for parotid RPA, to determine extension and detect subclinical lesions. Radiotherapy should be considered in case of multi-recurrent pleomorphic adenoma after macroscopically complete revision surgery at high risk of new recurrence (microscopic residual disease), in case of RPA after incomplete resection, and in non-operable RPA.

Cut-off values and significance of Oil Red O-positive cells in bronchoalveolar lavage fluid.

OBJECTIVE: To evaluate the percentage and predictive value of Oil Red O-positive macrophages (ORO-PM) to identify lipid-laden macrophages in bronchoalveolar lavage fluids (BALF) from patients with different pathologies. METHODS: The percentage and absolute numbers of ORO-PM were evaluated in 305 BALF. The patients were separated into ten groups: corticosteroid treatment (n = 18), amiodarone treatment (n = 8), interstitial fibrosis (n = 11), human immunodeficiency virus (HIV)-positive (n = 25), infectious pneumonia (n = 43), severe haematological disorder (n = 25), interstitial syndrome (n = 109), suspicion of cancer (n = 17), transplant recipients (n = 50) and controls (n = 43). The total and differential cell counts in BALF were recorded. The presence of specific pathogens was also noted. Parametric and non-parametric tests were used to compare the values between groups. Receiver-operating characteristics (ROC) curves were established in order to determine a cut-off value. RESULTS: The percentages of ORO-PM were (mean +/- standard deviation) 21.67 +/- 29.12 in the corticosteroid group, 10.00 +/- 12.49 in the amiodarone group, 19.45 +/- 20.72 in the interstitial fibrosis group, 47.80 +/- 30.46 in the HIV group, 19.72 +/- 26.26 in the infectious pneumonia group, 27.42 +/- 30.04 in the severe haematological disorder group, 25.18 +/- 30.63 in the interstitial syndrome group, 17.64 +/- 27.76 in the suspicion of cancer group, 22.50 +/- 27.27 in the transplanted recipients group and 2.63 +/- 3.48 in the control group. Significantly higher values were found in all groups when compared with the control group (P < 0.001). Only the HIV group showed higher numbers of ORO-PM when compared with the interstitial syndrome group (P < 0.01). According to ROC curves, > 6% ORO-PM was suggested as the positive cut-off value. CONCLUSION: Significantly increased numbers of ORO-PM were associated with various lung pathologies. However, the higher numbers observed in HIV patients require further investigations.

[Urologic aspects of Polyomavirus infection]. / Aspects urologiques de l'infection à Polyomavirus.

JC virus (JCV) and BK virus (BKV) are human Polyomaviruses of the papovavirus family, which also includes a simian vacuolating virus 40 (SV40). Human Polyomaviruses were first isolated in 1971 from the brain (JCV) and urine (BKV) of two different patients. Human Polyomaviruses have a limited and specific tissue tropism infecting the renal tubular cells, the urothelium, the B cells and the brain cells. The virus infects the majority of the human population with seroconversion occurring during adolescence. The detection of the virus may be cytological, pathological, virological or immunological. Following a typically subclinical primary infection, Polyomavirus establishes a life-long persistent infection, especially in the urinary tract. BKV is known to reactivate and cause severe disease in immunosuppressed patients. The presence of Polyomavirus outside conditions of immunosuppression raises the question of its meaning and its therapeutic management. Given the ubiquitous nature of the virus and its strong association with cancer in animal models, they may play an etiological role in human malignancies. Here, we describe the biology of human Polyomaviruses, review their non-malignant and malignant potentials, and discuss the therapeutic aspect.

An innovative in vitro device providing continuous low doses of γ-rays mimicking exposure to the space environment: A dosimetric study.

Astronauts are exposed to microgravity and chronic irradiation but experimental conditions combining these two factors are difficult to reproduce on earth. We have created an experimental device able to combine chronic irradiation and altered gravity that may be used for cell cultures or plant models in a ground based facility. Irradiation was provided by thorium nitrate powder, conditioned so as to constitute a sealed source that could be placed in an incubator. Cell plates or plant seedlings could be placed in direct contact with the source or at various distances above it. Moreover, a random positioning machine (RPM) could be positioned on the source to simulate microgravity. The activity of the source was established using the Bateman formula. The spectrum of the source, calculated according to the natural decrease of radioactivity and the gamma spectrometry, showed very good adequacy. The experimental fluence was close to the theoretical fluence evaluation, attesting its uniform distribution. A Monte Carlo model of the irradiation device was processed by GATE code. Dosimetry was performed with radiophotoluminescent dosimeters exposed for one month at different locations (x and y axes) in various cell culture conditions. Using the RPM placed on the source, we reached a mean absorbed dose of gamma rays of (0.33 ± 0.17) mSv per day. In conclusion, we have elaborated an innovative device allowing chronic radiation exposure to be combined with altered gravity. Given the limited access to the International Space Station, this device could be useful to researchers interested in the field of space biology.

Effect of continuous irradiation with a very low dose of gamma rays on life span and the immune system in SJL mice prone to B-cell lymphoma.

Ionizing radiation has been shown to have dose- and dose-rate-dependent carcinogenic effects on the hematopoietic and lymphoreticular systems. We report here that continuous exposure to a low dose of gamma rays influences the course of spontaneous B-cell lymphoma in SJL mice. We studied the biological effects of 10 cGy year(-1) gamma rays on the life span of 560 4-week-old SJL/J female mice and on various parameters of the cell-mediated immune response. Life span was slightly prolonged. The mean survival was 397 days for controls and 417 days for irradiated mice that died with lymphoma (P = 0.34). In lymph nodes and spleen, lower percentages of CD4+ and CD8+ T cells were observed in irradiated mice before 32 weeks. Interestingly, the percentages of CD49+ NK cells were increased in the spleens of irradiated mice at 28 weeks (0.61 +/- 0.08% compared to 0.43 +/- 0.12% in controls, P = 0.01) and at 32 weeks (0.62 +/- 0.24% compared to 0.33 +/- 0.09%, P = 0.02), while NK cell activity remained unchanged in exposed mice. These results provide further support for the absence of harmful effects of a continuous very low dose of radiation on life span and incidence of lymphoma in SJL mice.