Laser-assisted cryosurgery in ex vivo mice hepatic tissue: viability assays using green fluorescent protein.

An experimental investigation is carried out to develop a novel approach to cryosurgery, where laser heating counteracts tissue freezing to better confine damage to the targeted cancerous tissue within a lethal low-temperature isothermal boundary-an approach we refer to as laser-assisted cryosurgery (LAC). The advantage of this procedure relative to conventional cryosurgery assisted with urethral warmers or cryoheaters is that laser heating provides volumetric rather than superficial heating, which leads to deeper penetration, more homogeneous tissue protection and better demarcation of the destructive freezing effect to a well-defined targeted volume. Tissue viability assays are performed using green fluorescence protein (GFP) as a viability marker and correlated with temperature history after performing LAC procedures on ex vivo mice hepatic tissue. The limit for cell denaturation at the irradiated surface predicted by GFP analysis is further confirmed using reverse transcription polymerase chain reaction (RT-PCR). In addition, the correlation between GFP fluorescence and cell viability and loss of GFP fluorescence in non-viable cells has been tested and validated by histological analysis using a standard cell viability measuring method (hematoxylin and eosin staining). Analysis of our experimental measurements show that reproducible thermal gradients (of 236 °C/cm) and predictable tissue necrosis can be reliably produced by LAC without exceeding temperature thresholds for cell denaturation (of T (surf) ≈ 48 °C) beyond preset tissue boundaries (with resolution of 0.1 °C/mm). The results have shown the feasibility of controlling temperatures at specified tissue locations to prevent hyperthermal or freezing damage.

[Lobular intra-epithelial neoplasia: atypical lobular hyperplasia and lobular carcinoma in situ]. / Néoplasies lobulaires intra-épithéliales: hyperplasie lobulaire atypique et cancer lobulaire in situ.

OBJECTIVE: To review main knowledge about lobular intra-epithelial neoplasia with special interest for daily practice management. MAIN RESULTS: Intra-epithelial lobular neoplasias (ILN) are non invasive proliferations within the terminal ducto-lobular unit of monomorphic loosely cohesive small cells. A lack of expression of the E-cadherin adhesion molecule is often observed as in invasive lobular breast cancer. ILN are infrequent, however, a rise in incidence partly, due to the generalization of mammographic screening, is observed. Actually ILN are usually asymptomatic and diagnosed after breast biopsy for unspecified microcalcifications. ILN are associated with an increased risk of breast cancer that persists over 20 years after the initial diagnosis. The average risk is 4.2 % for the ipsilateral breast and 3,5 % for the controlateral breast. However, a great variability in the risk estimation is observed between the studies. There is no consensus on how to treat ILN. Surgical options have varied from biopsy to bilateral mastectomy. Current tendency is favouring lumpectomy.

Distribution of tritium labeled 12(S) hydroxy-eicosatetraenoic acid (12-HETE) in the rat.

The in vivo metabolism of 12-(S)-Hydroxy-eicosatetraenoic acid (12-HETE), the end-lipoxygenase product of arachidonic acid in platelets, has been investigated in the rat. Fifty microcuries of 5,6-[3H]-12-HETE (50 Ci/mmol) were injected to anesthetized rats and the radioactivity was followed in plasma. At the end of the experiment, various organs of the animal were removed and the radioactivity attached to them was determined. The label of the plasma plateaued to approximately one third of the initial radioactivity ten minutes after the injection. Among the various organs tested (brain, heart, intestine, kidney, liver, lungs, spleen, testis/uterus) the kidney was far the most active to accumulate 12-HETE and/or its labeled metabolites, and no radioactivity could be detected in urine during the course of the experiment. The analysis of lipid extracts from the various tissues revealed that 12-HETE was not accumulating in its unesterified form but was likely bound to phospholipids. We conclude that, although the label providing from the initial 12-HETE did not completely disappear from plasma, circulating 12-HETE cannot be considered as a circulating marker of cell activation.

Isotope derivative assay of human serum bile acids.

A new method for the selective determination of the main serum bile acids has been developed. Serum samples with added 14C-labeled bile acid were submitted to deproteinization, alkaline hydrolysis, methylation, and were then chromatographed on alumina before acetylation with 2 microliters of [3H]acetic anhydride. Excess reagent was eliminated by evaporation; elimination of residual tritiated contaminants and separation of the doubly labeled bile acid derivatives were obtained by thin-layer chromatography, column chromatography on Lipidex 5000, and crystallization. The sensitivity of the method is about 10 pmol of each bile acid. Analyses of seven sera with normal or elevated concentration of bile acids by the proposed method and gas-liquid chromatography showed a close correlation (r = 0.94; slope = 0.93).

Bile acid sulfates in serum bile acids determination.

Some bile acid sulfates were synthesized and characterized. The configuration of sulfate groups at C-3, C-7 and C-12 positions was confirmed by Nuclear Magnetic Resonance analysis. These sulfates were utilized in a study of their chemical behaviour in different analytical procedures currently used for serum bile acids determination. Procedures for bile acids extraction from serum with ethanol or Amberlite XAD-2 result in an important loss of the most polar sulfated bile acids. Complete separation of unsulfated from sulfated bile acids on Sephadex LH-20 is not achieved when deconjugation of the most polar bile acid sulfate is slow but does not produce artifacts. Enzymatic determination of bile acids gives positive response with some bile acid sulfates. The current procedures of serum bile acids determination are discussed in consideration of these results.

Lymphographic patterns in systemic mastocytosis with diffuse bone involvement and hematological signs.

Lymphography revealed the nature of diffuse lymph node involvement in systemic mastocytosis with diffuse bone involvement and hematological signs, similar to that seen in some malignant hematological diseases or in osteomyelosclerosis. The final stages of transformation into acute lymphoblastic leukemia must also be considered.

[The effect of estradiol benzoate on liver function, cholesterol metabolism and bile acids in the quail]. / Effet du benzoate d'oestradiol sur la fonction hépatique et sur le métabolisme du cholesterol et des acides biliaires chez la caille.

Immature female quails were treated for 6 days with estradiol benzoate at daily 0.01-, 0.02-, 0.01-, and 1-mg dosages. At the end of treatment, bile outflow, biliary cholesterol (CST), and bile acid (BA) secretory rates and liver, bile, and serum CST and BA levels were determined. Some quails were used to measure the ratio (R) of the rates of intravenously injected [2-14C]acetate radioactivity incorporation in cholic (C) and chenodeoxycholic (CDC) acids excreted in bile. The oestrogenic treatments at doses greater than 0.01 mg/day caused a marked disturbance in hepatic function and in CST and BA metabolism: they induced an increase in relative liver weight, liver CST stores, serum CST, C, CDC, and SGOT levels and in choleresis (respectively up to 56, 57, 650, 6000, 700, 42, and 235% increase at the daily 0.1-mg dosage) and they decreased bile total BA and CDC levels, bile and serum CDC to C level ratios, and R ratio (by 71, 82, 69, 84, and 58%, respectively). An increase in the bile salts independent fraction of bile was responsible for hypercholeresis, whether alone at low dosage or in conjunction with other factors at higher dosage. These results are compared with those obtained in mammals, particularly in the rat.

[Postprandial variations in serum bile acid levels in humans free of hepatic or intestinal diseases (author's transl)]. / Variations postprandiales des taux d'acides billaires sériques chez l'homme indemne de troubles hépatiques ou intestinaux.

The levels of unsulfated, free or conjugated cholic, deoxycholic and chenodeoxycholic acids were measured using gas chromatography in 39 humans free of hepatic or intestinal diseases before and 10, 60, 120 and 180 min after ingestion of a standard meal. The probable maximal levels were determined with an error risk lower than 0.05. In fasting subjects, the observed values are comparable with those obtained by other authors working with gas chromatography or radioimmunoassay. Meal ingestion does not influence in the same way the serum levels of the various bile acids: the chemodeoxycholic serum level rose significantly in all cases whereas cholic and deoxycholic serum levels rose only in two-thirds of observed subjects; 60 and 120 min after the meal for chenodeoxycholic acid, and only 60 min after the meal for cholic acid, the mean values are significantly higher than the fasting ones; 120 min after the meal, the chenodeoxycholic and total bile acid probable maximal levels (respectively 7.4 and 10.3 micrometer) are twice the fasting ones. The cholic to chenodeoxycholic serum level ratio is nearly always lower than 1 but may reach 3. On the basis of these results, the validity and efficacy of the exploration tests based on serum bile acid level determinations are discussed.

[Mondor's disease. A radioclinical diagnosis, concerning one case (author's transl)]. / La maladie de Mondor. Un diagnostic radioclinique à propos d'une observation

The authors present a typical case of Mondor's disease of the superior external quadrant of the breast. They discuss two specific points: --the absence of superior-internal venous drainage; --the radiological picture given by a thrombosed vein.