Assessment of cellular adequacy of fine needle aspiration biopsy for small solid renal tumors.

OBJECTIVE: Our aim was to evaluate the performance of fine needle aspiration (FNA) as a method for obtaining cellular material for the tissue diagnosis of small solid renal tumours. METHODS: Fifty-five patients with a small solid renal tumour (4 cm or smaller) underwent a FNA biopsy under computed tomography guidance. Direct smears and rinse of needle with Cytolyt® were prepared. The cellular adequacy was divided into three groups: satisfactory cellular material, scant cellular material and noncellular material. RESULTS: Postoperative pathological diagnosis showed 35 clear cell carcinomas, 10 papillary carcinomas, four chromophobe carcinomas, four oncocytomas, one adenoma and one metastatic carcinoma. A total of 21 (38.2%) FNA biopsies belonged to noncellular group. For clear cell carcinomas, the cellularity of satisfactory cellular group, scant cellular group and noncellular group was, respectively, seven (20.0%), nine (25.7%) and 19 (54.3%). Of the 10 papillary carcinomas, nine (90%) and all four chromophobe carcinomas (100%) had satisfactory cellular materials. The percentages of noncellular group among clear cell carcinoma, papillary carcinoma and chromophobe carcinoma were significantly different (P = 0.0020). Fifteen tumours were 2 cm or smaller and 40 tumours were larger than 2 cm. No significant difference was found in the percentage of noncellular material for the tumours 2 cm or less (40.0%) and for the tumours larger than 2 cm (37.5%; P > 0.05). CONCLUSIONS: The cellularity of FNA biopsy of small solid renal masses might correlate with the subtype of renal cancers. FNA biopsy was an effective method of tissue sampling for papillary and chromophobe renal carcinomas.

Intraoperative Near-Infrared Fluorescence Imaging using indocyanine green in colorectal carcinomatosis surgery: Proof of concept.

PURPOSES: This study assesses the value of using Intraoperative Near Infrared Fluorescence Imaging and Indocyanine green to detect colorectal carcinomatosis during oncological surgery. In colorectal carcinomatosis cancer, two of the most important prognostic factors are completeness of staging and completeness of cytoreductive surgery. Presently, intraoperative assessment of tumoral margins relies on palpation and visual inspection. The recent introduction of Near Infrared fluorescence image guidance provides new opportunities for surgical roles, particularly in cancer surgery. METHODS: The study was a non-randomized, monocentric, pilot "ex vivo" blinded clinical trial validated by the ethical committee of University Hospital of Saint Etienne. Ten patients with colorectal carcinomatosis cancer scheduled for cytoreductive surgery were included. Patients received 0.25 mg/kg of Indocyanine green intravenously 24 h before surgery. A Near Infrared camera was used to detect "ex-vivo" fluorescent lesions. RESULTS: There was no surgical mortality. Each analysis was done blindly. In a total of 88 lesions analyzed, 58 were classified by a pathologist as cancerous and 30 as non-cancerous. Among the 58 cancerous lesions, 42 were correctly classified by the Intraoperative Near-Infrared camera (sensitivity of 72.4%). Among the 30 non-cancerous lesions, 18 were correctly classified by the Intraoperative Near-Infrared camera (specificity of 60.0%). CONCLUSIONS: Near Infrared fluorescence imaging is a promising technique for intraoperative tumor identification. It could help the surgeon to determine resection margins and reduce the risk of locoregional recurrence.

Aluminum content of human semen: implications for semen quality.

A deterioration of human semen quality has been observed over recent decades. A possible explanation could be an increased exposure to environmental pollutants, including aluminum. Our aim was to measure the aluminum concentration in the semen of 62 patients and to carry out a preliminary evaluation on its impact on specific semen parameters. For each patient, semen analyses were performed according to WHO guidelines. A graphite furnace atomic absorption spectrometry method was used to determine semen aluminum concentration. A cytological analysis using an aluminum-specific fluor, lumogallion, was also performed. The mean aluminum concentration in human semen was 339 µg/L. Patients with oligozoospermia had a statistically higher aluminum concentration than others. No significant difference was observed for other semen parameters. Cytological analysis showed the presence of aluminum in spermatozoa. This study provided unequivocal evidence of high concentrations of aluminum in human semen and suggested possible implications for spermatogenesis and sperm count.

Testicular biodistribution of 450 nm fluorescent latex particles after intramuscular injection in mice.

The significant expansion in the use of nanoparticles and submicron particles during the last 20 years has led to increasing concern about their potential toxicity to humans and particularly their impact on male fertility. Currently, an insufficient number of studies have focused on the testicular biodistribution of particles. The aim of our study was to assess the distribution of 450 nm fluorescent particles in mouse testes after intramuscular injection. To this end, testes were removed from 5 groups of 3 mice each at 1 h (H1), 4 days (D4), 21 days (D21), 45 days (D45) and 90 days (D90) after the injection of 7.28 × 109 particles in the tibialis anterior muscles of each mouse. We examined histological sections from these samples by epifluorescence microscopy and confocal microscopy and identified testicular biodistribution of a small number of particles in groups H1, D4, D21, D45 and D90. Using CD11b immunostaining, we showed that particles were not carried into the testis by macrophages. The intratesticular repartition of particles mainly followed testicular vascularization. Finally, we found some particles in seminiferous tubules but could not determine if the blood-testis barrier was crossed.

Size of submicrometric and nanometric particles affect cellular uptake and biological activity of macrophages in vitro.

BACKGROUND: Micrometric and nanometric particles are increasingly used in different fields and may exhibit variable toxicity levels depending on their physicochemical characteristics. The aim of this study was to determine the impact of the size parameter on cellular uptake and biological activity, working with well-characterized fluorescent particles. We focused our attention on macrophages, the main target cells of the respiratory system responsible for the phagocytosis of the particles. METHODS: FITC fluorescent silica particles of variable submicronic sizes (850, 500, 250 and 150 nm) but with similar surface coating (COOH) were tailored and physico-chemically characterized. These particles were then incubated with the RAW 264.7 macrophage cell line. After microscopic observations (SEM, TEM, confocal), a quantitative evaluation of the uptake was carried out. Fluorescence detected after a quenching with trypan blue allows us to distinguish and quantify entirely engulfed fluorescent particles from those just adhering to the cell membrane. Finally, these data were compared to the in vitro toxicity assessed in terms of cell damage, inflammation and oxidative stress (evaluated by LDH release, TNF-α and ROS production respectively). RESULTS AND CONCLUSION: Particles were well characterized (fluorescence, size distribution, zeta potential, agglomeration and surface groups) and easily visualized after cellular uptake using confocal and electron microscopy. The number of internalized particles was precisely evaluated. Size was found to be an important parameter regarding particles uptake and in vitro toxicity but this latter strongly depends on the particles doses employed.

Intraperitoneal free cancer cells in non-gynaecological adenocarcinomas: a reproducibility study.

OBJECTIVE: In recent years, therapeutic approaches including cytoreductive surgery followed by intraperitoneal chemotherapy have proven effective in peritoneal carcinomatosis of colorectal origin. If cytology is to be used to include patients in aggressive treatment regimens, it is necessary to evaluate its performance, particularly in terms of specificity. The aim of this study was to assess interobserver agreement for the detection of intraperitoneal free cancer cells (IFCCs) in patients with non-gynaecological adenocarcinomas. METHODS: Over a 5-year period, 1223 patients were recruited in 19 French surgery departments. Peritoneal samples were examined in 14 dispersed pathology laboratories. Giemsa-stained slides were sent to a control reader blind to the previous diagnosis. Discordant cases, concordant positive results and a random selection of negative concordant cases were reviewed by a panel of seven cytopathologists. The 'final diagnosis' was that of the consensus meetings but took into account locally-processed slides. RESULTS: Gathering dubious cases with negative results, a 95.6% concordance was achieved between local readers and the control reader. IFCCs were ascertained by the panel in 85 cases (7.0%). Eight of 873 colorectal cancers cases viewed locally were falsely positive (0.9%). Radiotherapy and neoadjuvant therapy had no impact on the false-positive rate as assessed by final validation by the panel (P > 0.05). Samples initially considered as dubious were reclassified as negative by the panel in 24 of 25 cases (96.0%). CONCLUSIONS: The panel consensus allowed reclassification of most dubious/equivocal peritoneal cytology cases, whereas clearcut distinction between benign and malignant cases was correctly achieved in almost all cases.

Quantification of microsized fluorescent particles phagocytosis to a better knowledge of toxicity mechanisms.

BACKGROUND: The use of micro- or nanometric particles is in full expansion for the development of new technologies. These particles may exhibit variable toxicity levels depending on their physicochemical characteristics. We focused our attention on macrophages (MA), the main target cells of the respiratory system responsible for the phagocytosis of the particles. The quantification of the amount of phagocytosed particles seems to be a major element for a better knowledge of toxicity mechanisms. The aim of this study was to develop a quantitative evaluation of uptake using both flow cytometry (FCM) and confocal microscopy to distinguish entirely engulfed fluorescent microsized particles from those just adherent to the cell membrane and to compare these data to in vitro toxicity assessments. METHODS: Fluorescent particles of variable and well-characterised sizes and surface coatings were incubated with MA (RAW 264.7 cell line). Analyses were performed using confocal microscopy and FCM. The biological toxicity of the particles was evaluated [lactate dehydrogenase (LDH) release, tumor necrosis factor (TNF)-α, and reactive oxygen species (ROS) production]. RESULTS AND CONCLUSION: Confocal imaging allowed visualization of entirely engulfed beads. The amount of phagocytic cells was greater for carboxylate 2-µm beads (49 ± 11%) than for amine 1-µm beads (18 ± 5%). Similarly, side scatter geometric means, reflecting cellular complexity, were 446 ± 7 and 139 ± 12, respectively. These results confirm that the phagocytosis level highly depends on the size and surface chemical groups of the particles. Only TNF-α and global ROS production varied significantly after 24-h incubation. There was no effect on LDH and H(2)O(2) production.

[Results of fine-needle aspiration cytology (FNAC) in the diagnosis of indeterminate solid renal tumours]. / Résultats de l'aspiration cytologique à l'aiguille fine dans le diagnostic des tumeurs solides du rein de nature indéterminée.

OBJECTIVE: To evaluate the accuracy of fine-needle aspiration cytology for the diagnosis of imaging indeterminate solid renal tumours. MATERIALS AND METHODS: From February 2003 to February 2009, 60 cytoaspirations have been performed to 20 female and 40 male patients (average age: 62.0+/-14.2 years) with an indeterminate solid renal mass by imaging. The average tumour size was 3.4+/-2.8 cm. The cytoaspiration was performed through a 22 Gauge needle under CT (n=39) or US (n=21) guidance. Papanicolaou staining was used. All slides were examined by one experienced cytologist without any clinicoradiological information. The results were given as malignant, benign, suspect or non significant. A classification of subtypes of renal cancer might be added by the cytologist. RESULTS: Twenty-one cytoaspirations (35%) were non significant while 39 (65%) showed cells of interest. Among these 39 cellular cytoaspirations, the specificity for malignancy or benignity was 89.7%. The proportion of non-significant samples was the same in tumors lesser than 2 cm (38.4%) as in tumors 2-4 cm (38.8%) (p=1.000, Fisher's exact test). Subtype identification was only reliable for clear cell carcinomas. No complication was observed. CONCLUSION: The fine-needle aspiration cytology is an auxiliary technique for the diagnosis of indeterminate solid renal tumours. This simple and mini-invasive technique had a high specificity but a low sensitivity in our experience. Fine-needle aspiration is complementary to core biopsy which remains the gold standard of percutaneous sampling.

MN/CA9: a potential gene marker for detection of malignant cells in effusions.

Many cancers cause malignant effusions. The presence of malignant cells in effusions has implications in diagnosis, tumour staging and prognosis. The detection of malignant cells currently presents a challenge for cytopathologists. New adjunctive methods are needed. Although the effusions provide excellent materials for molecular assay, the available molecular markers are extremely limited, which hinders its clinical application. MN/CA9 has proved to be a valuable marker in many cancers such as lung, breast, colon, kidney, etc. The present study was to evaluate MN/CA9 as a new molecular marker for the detection of cancer cells in pleural effusions. Seventy-one pleural effusions including 59 malignant effusions from patients with cancer, and 12 patients with benign diseases as a control, were subjected to RT-PCR for detection of MN/CA9 gene expression. MN/CA9 gene expression was detected in 53/59 (89.8%) pleural effusions from cancer patients (15/16 for breast cancers, 10/11 for lung cancers, 4/4 for ovary cancers, 2/3 for colon-rectal cancers, 5/6 for cancers of unknown site, 7/8 for mesothelioma and 10/11 for other cancers). Furthermore, MN/CA9 was positive in 13/18 (72.2%) of cytologically negative effusions of cancer patients. MN/CA9 was detected in only 1/12 (8.3%) effusions from the control patients (p < 0.01). The sensitivity and specificity of MN/CA9 gene expression were, respectively, 89.8% and 91.7%. Our preliminary results suggest that MN/CA9 could be a potential marker for the detection of malignant cells in effusions. A large-scale study is needed to confirm these results.

Cost efficiency analysis of modern cytocentrifugation methods versus liquid based (Cytyc Thinprep) processing of urinary samples.

BACKGROUND/AIMS: Liquid based cytology (LBC) was developed as a replacement for cytocentrifugation in the treatment of cell suspensions. Because accurate data comparing the quality and total cost of modern cytocentrifugation methods versus LBC in non-gynaecological samples are not available, this study was designed to investigate these issues. METHODS: The study comprised 224 urine samples treated with the Thermo Shandon Cytospin 4 using reusable TPX chambers, disposable Cytofunnels for samples up to 0.5 ml, and disposable Megafunnels for samples up to 6 ml. Each method was compared with the Cytyc Thinprep processing of a paired sample. Quality was assessed by scoring cellularity, fixation, red blood cells, leucocytes, abnormalities of urothelial cells, and suitability for molecular studies. Wage costs, investment, and consumables allowed a "total cost" to be calculated on the basis of 200 specimens/month. Total cost and quality combined were used to calculate an index of total quality (ITQ). RESULTS: Cytocentrifugation with disposable chambers resulted in a global quality superior to that of Cytyc Thinprep LBC. Preparation and screening times were 2.25 and 1.33-2 times greater when using LBC compared with cytocentrifugation. The total cost each month reached 1960.23 $ to 2833.43 $ for cytocentrifugation methods and 5464.95 $ for Cytyc Thinprep LBC (92.8-178.8% increased cost). ITQ of cytocentrifugation with disposable chambers surpassed that of Cytyc Thinprep LBC (37.25/32.08 and 9.98, respectively). CONCLUSION: Cytyc Thinprep LBC and cytocentrifugation are both appropriate methods for cytology based molecular studies, but cytocentrifugation remains the quality standard for current treatment of urinary samples because of its lower cost.

Prospective flow cytometric DNA analysis of hepatocellular carcinoma specimens collected by ultrasound-guided fine needle aspiration.

BACKGROUND: The survival of 52 patients with hepatocellular carcinoma (HCC) seen during the last 4 years was analyzed prospectively on the basis of disease stage and nuclear DNA content. METHODS: Ploidy was measured by flow cytometry (FCM). Cells for cytologic diagnosis and FCM were collected by ultrasound-guided fine needle aspiration. RESULTS: DNA aneuploidy, which was detected in 62% of the patients, did not correlate with clinicopathologic features, except in the sonographic aspect (P = 0.03). However, ploidy correlated significantly with survival; the survival times for patients with an aneuploid DNA index were significantly shorter than for those with a diploid index (P = 0.02). In a Cox multivariate analysis, DNA content was prognostically significant, as were the grade of cirrhosis severity and the echographic aspect. CONCLUSIONS: In addition to the clinicopathologic features observed, FCM DNA analysis of ultrasound-guided fine needle aspirates from HCC is a simple and valid method for estimating a prognosis of these patients.

Flow cytometric analysis of DNA content in colorectal polyps resected at endoscopy.

Samples from 64 consecutive resected colorectal polyps were preserved in liquid nitrogen and analyzed by flow cytometry to assess the nuclear DNA content, which was compared to the histopathologic findings. The frequency of aneuploidy in these nonselected polyps was 17.2%. There was a significant correlation between the diameter of the polyp and the frequency of aneuploidy (P = .04), with all aneuploid polyps being greater than or equal to 10 mm. Similarly, aneuploidy was significantly more frequent for polyps that were both greater than or equal 20 mm and showed at least severe dysplasia (P = .02). On the other hand, there was no correlation between the ploidy and the gross histopathologic type (tubular, tubulovillous or villous), and the proliferation index did not correlate with any of the parameters studied.

[Value of the peroperative association of echography, extemporaneous cytology and immediately available insulin blood levels of the portal vein. Apropos of 2 cases of insulinoma]. / Intérêt de l'association per-opératoire de l'échographie, de la cytologie extemporanée et des dosages d'insulinémie portale avec résultats immédiats. A propos de deux cas d'insulinome.

The authors report two cases of insulinoma managed by per-operative ultrasound and cytology, together with immediate portal vein insulin levels. They stress the value of these per-operative exploratory techniques which reduce the number of pre-operative invasive examinations while ensuring a greater efficacy in the localization and treatment of these tumors.

Bronchial carcinoma and image analysis. Differentiation between poorly differentiated epidermoid carcinoma and small-cell carcinoma.

The utility of automated image analysis in the distinction between poorly differentiated epidermoid carcinoma (eight cases) and small-cell carcinoma (ten cases) was studied. Material obtained using the bronchial brushing technique was prepared by a cytocentrifugation technique. In each case, a total of 100 bronchial cell nuclei were selected using the Leitz TAS, which measured eight parameters per cell in order to ascertain the homogeneity or the heterogeneity of the nuclear populations. Except for one sample exhibiting preparation artifacts, the method proved capable of differentiating between these two types of bronchial carcinoma, with heterogeneity of the malignant nuclei indicating an epidermoid carcinoma and homogeneity indicating a small-cell carcinoma. A correlation was observed to exist between the morphologic and the morphometric criteria.

Radio-anatomic study of the carotid axis with regard to the implantation of microsurgical vascular anastomoses.

The authors report a radio-anatomic study of the carotid axis in the context of the implantation of microsurgical vascular anastomoses. The information derived from 36 dissections and 50 arteriographic studies suggests the preferential use of the superior thyroid, facial and lingual branches. Section and turning-down of the external carotid and direct end-to-side implantation on the common carotid could provide fall-back solutions when the collaterals are slender. Because of anatomic variations and frequent atheromatous lesions, arteriographic study before implantation appears essential.

[Post-anesthetic hepatitis. The role of halothane and antimitotic combinations]. / Hépatite postanesthésique. Responsabilité de l'association halothane-antimitotiques.

A 12 year old boy with Burkitt's lymphoma developed severe hepatitis with hepatomegaly, subclinical jaundice, and a small rise in body temperature, associated with an important rise in SGPT and fall in prothrombin titres, 6 days after anticancer chemotherapy and 24 hours after halothane anaesthesia. Hepatitis A and B serology remained negative. This hepatic failure explained perhaps the unusually severe vincristine toxicity which gave rise to a polyneuritis with important sequelae. The association of halothane hepatitis with antimitotic drugs appeared particularly dangerous, and halothane should probably be avoided in all patients been given or about to be given anticancer chemotherapy.