The clinical impact of circulating tumor cell (CTC) detection is controversial, mainly due to drawbacks of molecular approaches applied to this field. We sought to determine if the specific identification and counting of circulating tumor cells by cytomorphologic analysis has clinical usefulness. Peripheral blood (6 mL), treated using isolation by size of epithelial tumor cells, was obtained from 44 patients with primary liver cancer (PLC) and without metastases, 30 patients with chronic active hepatitis, 39 with liver cirrhosis, and 38 healthy individuals, and followed up for a mean period of 1 year. We searched for beta-catenin mutations in 60 single microdissected CTCs. One patient with liver cancer developed extrahepatic metastases during follow-up. CTCs and microemboli were found in 23 of the 44 patients with liver cancer and in none of the patients with chronic active hepatitis, patients with cirrhosis, or healthy subjects. Their presence was significantly associated with tumor diffusion (P =.0001) and portal tumor thrombosis (P =.006). Both the presence (P =.01) and number (P =.02) of CTCs and microemboli were significantly associated with a shorter survival. Beta-catenin mutations were found in 3 of 60 CTCs, arguing against their impact on the initial step of tumor cell invasion. In conclusion, the highly sensitive and specific detection of CTCs and microemboli may have clinical implications for cancer staging and outcome prediction. We also show the feasibility of molecular studies of individual circulating tumor cells, aimed at identifying gene mutations involved in tumor invasion.

Background and objective: Some halogenated agents, especially methoxyflurane, because of a higher level of fluoride production, induce a renal concentrating defect that could be related to an ascending limb impairment. We investigated the mechanisms of fluoride toxicity on an immortalized cell line.Methods: Cells were cultured for 2, 6 or 24 h in the presence of fluoride. Toxicity evaluation was based on: cell numbers, protein content, leucine-incorporation, lactate dehydrogenase (LDH) and N-acetyl-beta-glucosaminidase (NAG) releases, Na-K-ATPase and Na-K-2Cl activities, electron microscope studies. Infrared analysis and fluoride microdetermination allowed crystal components.Results: At 5 mmol after 24 h, fluoride decreased cell numbers (-14%, *P < 0.05), protein content (-16%*), leucine incorporation (-54%), Na-K-2Cl activity (-84%), increased LDH (+145%) and NAG release (+ 190%). Na-K-ATPase was more sensitive and impaired from 1 mmol for 24 h and after 2 h at 5 mmol. Crystal formation in mitochondria occurred after 6 h at 5 mmol. Infra-red analysis and fluoride microdetermination established that crystals contained sodium, phosphate and fluoride.Conclusions: The results suggest that the Na-K-ATPase pump is a major target for fluoride toxicity in Henle's loop.

Une frequence non negligeable : Les lithiases medicamenteuses representent 1,5 a 2% des calculs de l'appareil urinaire. Les medicaments peuvent etre physiquement presents dans les calculs en raison de leur faible solubilite dans l'urine ou inducteurs de lithiase par leurs effets metaboliques. Ils peuvent etre responsables de l'apparition des calculs ou contribuent simplement a la croissance de calculs preexistants. Principaux medicaments impliques : Ce sont le triamterene, la glafenine, le piridoxilate, la sulfadiazine et l'indinavir, mais aussi les supplementations calciques et vitaminiques D mal controlees. D'autres medicaments contribuent occasionnellement a la formation de calculs, en particulier quelques antibiotiques et antiseptiques urinaires. . Diagnostic et prevention : Aujourd'hui, on peut distinguer des lithiases medicamenteuses qui ont disparu (glafenine, piridoxilate), des lithiases qui sont reapparues (sulfadiazine) et de nouvelles lithiases medicamenteuses (ceftriaxone, amoxicilline, silice opaline chez le jeune enfant, indinavir). L'analyse physique des calculs par des methodes fiables notamment la spectrophotometrie infrarouge et l'interrogatoire du patient pour reconstituer son passe therapeutique sont les elements-cles du diagnostic. Il existe des facteurs de risque specifiques au developpement de certaines lithiases medicamenteuses. La prise en consideration de ces facteurs lors de la prescription du medicament et leur surveillance lors des traitement au long cours devraient permettre de reduire la frequence des complications lithiasiques. . Au point de vue therapeutique : Toutes les lithiases radiotransparentes ne sont pas solubles en milieu alcalin. En particulier, les lithiases medicamenteuses d'indinavir ou celles d'urate d'ammonium induites par les laxatifs ne peuvent pas etre dissoutes par une cure de diurese alcaline.

A 24-year-old woman was treated with zidovudine, lamivudine, characteristic pattern; (5) long-term renal dysfunction, if any, is and indinavir because of a needle-stick injury. Baseline laboratory unknown, especially in HIV-negative individuals; and (6) indievaluation revealed the following: serology for antibodies to HIV navir-induced crystalluria can be diagnosed in the office with use was negative, a hepatitis panel was negative, and a rapid plasma of light microscopy. reagin (RPR) test was negative; microscopic evaluation of a urine

Primary hyperoxaluria (PH) is a severe inherited disease induced by an enzymatic deficiency responsible for high endogenous production of oxalate. Oxalate ions are excreted by the kidney where they can form an insoluble salt with calcium ions, thus inducing urinary stones, crystal deposition in the tubular lumen and renal parenchyma leading to nephrocalcinosis and renal failure. Eighty-seven calculi from 63 PH patients with primary hyperoxaluria were analyzed and compared to 24,130 calculi from unselected consecutive stone formers referred to our laboratory between January 1977 and December 1996. All stones were analyzed according to a protocol including morphological examination of both surface and cross-section, and sequential infrared identification of the crystalline phases. A typical aspect of both surface and section corresponding to morphological type Ic according to our proposed classification (Daudon et al. Scanning Microsc 1993, 7:1081-1106) was observed in all patients but two whereas only two type Ic stones were observed among patients without primary hyperoxaluria. The latter two patients suffered from severe inflammatory bowel disease and developed heavy hyperoxaluria following extensive ileal resection. We conclude that evidence of type Ic morphology is a simple, cheap and fast tool to detect diseases with heavy hyperoxaluria such as primary hyperoxaluria.

OBJECTIVES: Anti-proteases, a new class of Anti-HIV drugs used in combination with reverse transcriptase inhibitors have led to spectacular improvement in the patients' clinical status. Since April 1996, indinavir is the most widely prescribed anti-protease in France. PATIENTS AND METHODS: From July 1996 to July 1997, we analyzed 46 spontaneously expulsed stones in 45 HIV+ patients (35 men and 10 women; age range 25 to 64 years) given indinavir in combination viith other drugs since one week to ten months. Only six patients were known to have a past history of renal lithiasis. RESULTS: Forty-one calculi contained indinavir monohydrate (INDM) identified by mass spectrometry and infrared spectrophotometry. INDM was the only component excepting proteins in 39/45 calculi. in the 12 others, other compounds were also identified. Among the 114 urine samples collected 2 to 3 hours after an 800 mg dose of indinavir, 38 (33%) monohydrate indinavir crystals, identified by infrared microscopy. Mean urinary pH was significantly higher than in samples without INDM crystals (6.53 +/- 0.68 versus 5.96 +/- 0.71, p < 0.001). CONCLUSION: Two measures could possibly reduce the risk of crystalization: administration of urine acidifiers and increased fluid intake to raise diuresis. Alkalinisation is not indicated. Long-term increased fluid intake should be prefered over acidification which could be reserved solely for the treatment of drug-induced lithiasis.

Numerous studies of calcium oxalate crystal formation have been carried out in the past two decades. In the present study, experiments were carried out to validate a turbidimetric method allowing to assess the calcium oxalate crystallization process. This method is quick and reproducible and can be used to quantify the inhibition of calcium oxalate crystal growth by various compounds. An experimental method of validation has been developed, which consisted in filtering solutions pure or containing modifiers at given crystallization times, photographing the filters used on scanning electron microscopy and analyzing the images using mathematical methodology. The results obtained through image analysis, namely crystal density (mean particle number per unit volume) and mean area, were correlated with the turbidimetric parameters. This finding was consistent with the qualitative examination of the photographs. Moreover, the morphological differences in crystals observed on the photographs were confirmed by the calculated length/width ratio. One can therefore assume that inhibition of calcium oxalate crystal growth is at least, partly explained by surface adsorption phenomena, which may add to complex formation.

Crystal-induced kidney disease is a frequent occurrence in human pathology. It is becoming more and more apparent that knowledge of kidney stone composition and structure appears to be the key for establishing the etiology of stone disease. A number of analytical methods may be applied to stone analysis, but only a few of them are able to quickly and easily provide extensive information on both stone structure and composition relevant for clinical diagnosis. More than 12,000 calculi were analyzed using a combination of microscopic examination, sequential infrared (IR) analysis by Fourier transform infrared spectroscopy (FTIR) of each part of stone, and quantification of all components present. We also investigated 50 biopsies using FTIR microscopy. Our results confirm that IR spectroscopy is a reliable and accurate technique for both molecular and crystalline identification. Some limitations of standard procedures, because of very small samples or due to absorption band overlap, can be solved using FTIR micromethod or a particular method like IR microscopy. In such cases, the spectrum identification must be conducted in different manners. Until now, spectral identification procedures based on computerized spectra libraries must be used with caution because of false results, mainly for mixtures of mineral compounds. Trained eyes always provide the best results for reading spectra from common stones. In routine practice, accurate identification of all components present in calculi is necessary for understanding urolithiasis mechanisms, but only semiquantitative assessment is sufficient to guide physicians toward establishing correct etiology. © 1997 John Wiley & Sons, Inc. Biospectroscopy 3: 347–369, 1997