[Acantholytic dermatosis in patients treated by vemurafenib: 2 cases]. / Deux cas de dermatose acantholytique sous vémurafénib.

BACKGROUND: Acantholytic dyskeratosis under BRAF inhibitors are dermatological diseases rarely reported to date. PATIENTS AND METHODS: We report 2 cases of acantholytic dyskeratosis, reaching the trunk and the seborrheic zones, not itchy, appeared one month after the introduction of vemurafenib. The histological analysis was typical of a "Grover-like rash" for the 2 patients. DISCUSSION: The appearance of acantholytic dyskeratosis under vemurafenib, a BRAF inhibitor, seems related with a paradoxical activation of the MAP-kinases pathway and with a growth acceleration of lesions in which RAS mutations of keratinocytes. Theses dermatoses seem also to occur with dabrafenib. CONCLUSION: The patients treated by BRAF inhibitors (vemurafenib and dabrafenib) can present acantholytic dyskeratosis. The arisen of this mild dermatosis does not question, of course, the continuation of the treatment. These cutaneous manifestations can be managed with emollients.

Transcultural validation of the Risk Assessment and Predictor Tool (RAPT) to predict discharge outcomes after total hip replacement.

OBJECTIVE: To assess the relevance of the RAPT (Risk Assessment and Prediction Tool), among a cohort of patients undergoing total hip arthroplasty (THA). METHOD: Prospective study of a cohort of patients evaluated before and after THA. The difference between the postoperative orientation predicted by the RAPT and the real one is assessed. Clinical, environmental and psychosocial criteria that could significantly change the orientation are discussed. RESULTS: One hundred and thirty-four patients (94 women and 40 men) were included. The average age was 71.6 (±10) years. Primary hip osteoarthritis was the indication for surgery in 78% of cases. The average length of stay in the surgery ward was 10 (±3) days. It was significantly higher for patients referred to a rehabilitation ward (P<0.0001). Sixty-six percent of patients were referred to a rehabilitation ward and 34% returned directly home. The average length of stay in rehabilitation ward was 27 (±13) days. The validity of the RAPT as a help decision tool has been confirmed. Thus, a low RAPT score was significantly associated with more frequent referral to a rehabilitation ward, conversely, a high RAPT score is significantly related to more frequent direct return to home. CONCLUSION: This study confirmed the usefulness of the RAPT to help in patient orientation decision after total hip arthroplasty. The patient preference remains the main variable for orientation after THA. By the way, the patient preference must not be integrated into the RAPT, but need to be collected and be discussed with the patient.

Management of pain associated with debridement of leg ulcers: a randomized, multicentre, pilot study comparing nitrous oxide-oxygen mixture inhalation and lidocaïne-prilocaïne cream.

BACKGROUND: Mechanical debridement of fibrin and/or necrosis promotes healing of arterial and venous leg ulcers but is limited by pain associated with the procedure. OBJECTIVE: The main objective of this study was to compare the respective analgesic effect of nitrous oxide oxygen mixture (NOOM) inhalation and lidocaïne-prilocaïne cream (LPC) application during the mechanical repeated debridement of chronic arterial and venous leg ulcers. METHODS: In this randomized, multicentre, open-label study, pain was evaluated before and after each care and debridement session using a Visual Analog Scale (VAS) and a Verbal Rating Scale (VRS), in the context of usual debridement and wound care process. The Quality of debridement and tolerability of the treatments were also assessed. RESULTS: Forty-one patients were randomized: 20 received NOOM and 21 LPC. Pain assessed by VAS and VRS was more intense in the NOOM group than in the LPC group (5.29 vs. 3.68 and 2.87 vs. 1.71, P<0.001, for the two scales respectively). No differences were found concerning quality of debridement, safety or tolerability between the two groups. CONCLUSION: This pilot study demonstrates the superiority of the LPC over NOOM for pain control during the mechanical debridement of chronic leg ulcers.

Diplopia as an initial manifestation of disseminated non-hodgkin's lymphoma.

A 37-year-old male presented a sudden diplopia. Ocular examination showed a partial paresis of the left eye (LE) on the left gaze, progressing in a few days towards a total ophthalmoplegia. Further investigation revealed an intra-orbital mass, immunohistologically diagnosed as a Diffuse Large B-cell Lymphoma (DLBCL), according to the WHO classification. Since the patient was in an advanced, disseminated stage of the disease (IVA-E), treatment was based on systemic and intrathecal chemotherapy with a pancranial radiotherapy. The clinical course was poor with only a 10-month survival. We wish to stress that the possibility of orbital malignancy in young adults with acute onset of ophthalmoplegia should be included in the differential diagnosis.

Cerebrospinal fluid flow waveforms: MR analysis in chronic adult hydrocephalus.

UNLABELLED: Henry-Feugeas MC, Idy-Peretti I, Baledent O, et al. Cerebrospinal fluid flow waveforms: MR analysis in chronic adult hydrocephalus. Invest Radiol 2001;36:146-154. RATIONALE AND OBJECTIVES: To analyze changes in cerebrospinal fluid (CSF) hydrodynamics in chronic adult hydrocephalus. METHODS: Phase-contrast cine-MR acquisitions were used to explore the ventricular system and the upper ventral cervical spaces of 16 patients. The aqueductal jet was explored in 32 control subjects. RESULTS: The duration of pulsatile caudal CSF flow (ie, CSF systole) was abnormally short in patients with active idiopathic and obstructive hydrocephalus. The duration of CSF cervical systole was normal in patients with stable hydrocephalus. The aqueductal stroke volume could be increased in stable communicating hydrocephalus. Patients who responded to shunting had shortened CSF systoles and hyperpulsatile ventricular patterns. Successful CSF diversion resulted in longer CSF systoles and CSF ventricular patterns that were no longer hyperpulsatile. CONCLUSIONS: Magnetic resonance analysis of CSF flow can show craniospinal dissociation and limitation of CSF outflow from the ventricles in both obstructive and communicating hydrocephalus; it should help determine the response to shunting in communicating hydrocephalus.

DNA-Dependent protein kinase is not required for efficient lentivirus integration.

How DNA is repaired after retrovirus integration is not well understood. DNA-dependent protein kinase (DNA-PK) is known to play a central role in the repair of double-stranded DNA breaks. Recently, a role for DNA-PK in retroviral DNA integration has been proposed (R. Daniel, R. A. Katz, and A. M. Skalka, Science 284:644-647, 1999). Reduced transduction efficiency and increased cell death by apoptosis were observed upon retrovirus infection of cultured scid cells. We have used a human immunodeficiency virus (HIV) type 1 (HIV-1)-derived lentivirus vector system to further investigate the role of DNA-PK during integration. We measured lentivirus transduction of scid mouse embryonic fibroblasts (MEF) and xrs-5 or xrs-6 cells. These cells are deficient in the catalytic subunit of DNA-PK and in Ku, the DNA-binding subunit of DNA-PK, respectively. At low vector titers, efficient and stable lentivirus transduction was obtained, excluding an essential role for DNA-PK in lentivirus integration. Likewise, the efficiency of transduction of HIV-derived vectors in scid mouse brain was as efficient as that in control mice, without evidence of apoptosis. We observed increased cell death in scid MEF and xrs-5 or xrs-6 cells, but only after transduction with high vector titers (multiplicity of infection [MOI], >1 transducing unit [TU]/cell) and subsequent passage of the transduced cells. At an MOI of <1 TU/cell, however, transduction efficiency was even higher in DNA-PK-deficient cells than in control cells. Taken together, the data suggest a protective role of DNA-PK against cellular toxicity induced by high levels of retrovirus integrase or integration. Another candidate cellular enzyme that has been claimed to play an important role during retrovirus integration is poly(ADP-ribose) polymerase (PARP). However, no inhibition of lentivirus vector-mediated transduction or HIV-1 replication by 3-methoxybenzamide, a known PARP inhibitor, was observed. In conclusion, DNA-PK and PARP are not essential for lentivirus integration.

High-level expression of active HIV-1 integrase from a synthetic gene in human cells.

A synthetic gene encoding for HIV-1 integrase was designed to circumvent the intrinsic instability and the repressor elements present in the wild-type gene. High-level expression of HIV-1 integrase was obtained in various human cell lines independently of viral accessory proteins. A human 293T cell line was selected that stably expresses HIV-1 integrase and has growth kinetics comparable to the parental cell line. The enzyme was localized in the nucleus and remained stably associated with the chromosomes during mitosis. Lentiviral vector particles carrying the inactivating D64V mutation in the integrase gene were capable of stably transducing 293T cells when complemented in the producer cells with integrase expressed from the synthetic gene. When the cell line that stably expresses integrase was infected with the defective viral particles, complementation of integrase activity was detected as well. Expression of active HIV-1 integrase in human cells will facilitate the study of the interplay between host and viral factors during integration.

Automated analysis of muscle fibre images.

A technique for studying the distribution and size of different fibre types in muscles is proposed for automated analysis of individual fibres in optical density images from ATPase-stained muscle sections. After delineation, fibres may be classified into different histological types (1, 2A, 2B and 2C) using the measurement of their mean optical density (mOD). The densitometric measurements were obtained from three serial histological slides stained under different conditions. The delineation procedure is performed on one of the images: the resulting mask is fitted to the other images using a linear coordinate transform. Along with densitometric measurements, the lesser diameter of the fibres is computed. Both in processing and in analysis, extensive use was made of mathematical morphology tools. All software was implemented on a VICOM digital image processor, extended with a VISIOMORPH morphoprocessor board.

Assay for acetyl-CoA:arylamine N-acetyltransferase by high-performance liquid chromatography applied to serotonin N-acetylation.

A specific assay to measure the activity of the enzyme acetyl-CoA:arylamine N-acetyltransferase (EC 2.3.1.5) from pigeon liver is described. The assay is based on the HPLC analysis of N-acetylserotonin formed by the enzymatic reaction. A reversed-phase column (Spherisorb 5-microns ODS 2; 150 x 3.2 mm) eluted with 0.1 M sodium acetate (pH 4.75)/methanol (75:25) permits baseline separation of serotonin and N-acetylserotonin within 5.3 min. Several variables on the enzyme reaction were studied to obtain maximum activity. The enzyme is most active in glycine buffer at pH 9.5. The apparent Km value for serotonin (at 0.6 mM CoASAc) is 0.246 mM and 9.9 microM for CoASAc (at 1.5 mM serotonin). To avoid acetyl-CoA or N-acetylserotonin consumption in side-reactions, the enzyme was purified. A two-step purification process (ammonium sulfate fractionation and affinity chromatography on immobilised amethopterin) yielded 60-70% of the initial enzyme activity with a purification factor of 455-560.

Transcriptional pattern of 21-hydroxylase gene (P-450C21) during embryonic development, before, and after birth in mice as determined by in situ hybridization.

21-Hydroxylase is a member of the P-450 superfamily of genes involved in the biosynthesis of cortisol and aldosterone in the adrenal cortex. Congenital adrenal hyperplasia, a well-characterized disease, originates from a lack of this enzyme. We present in this report an in situ hybridization study aimed at detecting 21-hydroxylase activity during murine development, from mid gestation to adulthood. Our results demonstrate that even during the embryonic period the adrenal cortex is the only major site of transcription of this enzyme, which is detectable beginning at embryonic day 14. In addition, a peculiar topographical pattern of transcriptional activity, characteristic of the stage of differentiation of the gland, could be drawn. Using a computer-assisted method, we were able to quantitate the relative transcription level at each stage of development. A steady increase in the level of transcription was demonstrated throughout embryonic life to birth, with a drop during the prepubertal period and a final rise at adult age. The possible physiological significance of our findings is discussed.

Fully automated measurements by light microscopy of tissue sections using a cellular array computer.

Software was developed for the acquisition, segmentation and analysis of microscopic OD-images on a VICOM digital image processor, extended with a VISIOMORPH morphoprocessor board. The delineation algorithms for peroxisomes, lysosomes, and nuclei in liver, kidney, and adrenal gland sections start by thresholding the difference between the original image and a low pass filtered version. The resulting binary mask is then processed by morphological operations in order to produce an object overlay. The efficiency of the programs is evaluated by comparing delineated objects at different OD-levels, created by varying the stain or by multiplying the original pixel values with constant factors. Manual delineation on some images is also used as a reference. More complex algorithms are used for the delineation of muscle fibres in ATP-ase-stained sections and immunocytochemically labelled cells in monolayer preparations. Muscle images from parallel sections with different stainings are matched with a coordinate transform, enabling the transfer of the object mask from a single delineated image to the unprocessed images and thus obtain all necessary information for fibre classification. After segmentation, the OD-images and their object overlays are fed into a data extraction program, measuring for each delineated object user-selected features. Data are sent to a VAX for statistical interpretation.

Prolactin cell subpopulations separated on discontinuous Percoll gradient: an immunocytochemical, biochemical, and physiological characterization.

A single-step procedure was devised to separate PRL cells from the rat anterior pituitary gland. After dissociation, cells were centrifuged on a Percoll gradient. Three layers were recovered. The composition of the different layers was evaluated using immunocytochemistry (with antisera to the six pituitary hormones), and in situ hybridization [with DNA complementary to PRL or to GH messenger RNA (mRNA)]. Both methods yielded identical values. PRL cells were recovered in the lower density layer (layer 1) with a good yield (that is 81% of the total PRL cells of the initial cell suspension) and in addition, markedly enriched (indeed 85% of the cells in layer 1 stained for PRL). A second layer (layer 2: intermediate density) contained most of the remaining PRL cells which were, however, heavily contaminated mainly by GH cells and cells that did not stain for any of the known pituitary hormones. A third layer (layer 3: higher density) was enriched in GH cells to 93% (representing, however, only 10% of the initial pituitary GH cells). In addition, PRL and GH were measured by RIA in culture medium and in cell lysates. Hormone biosynthesis was monitored by polyacrylamide gel electrophoresis and autoradiography after culture in the presence of [35S]methionine. These experiments confirmed that layer 1 was enriched in cells containing, and producing, PRL and depleted from GH cells. Cells in layer 2 contained and produced more GH than PRL. PRL cells from layer 1 responded to dopamine and to vasoactive intestinal polypeptide in the same way as PRL cells in the unseparated pituitary cell population. In contrast PRL cells in layer 2 had a lower basal secretion rate but a higher response to vasoactive intestinal polypeptide. Unless this represents a paracrine effect of non-PRL cells, PRL cells in layer 2 exhibit different properties and may therefore form a distinct subpopulation of PRL cells.