A new Monte Carlo model of a Cyberknife® system for the precise determination of out-of-field doses.

In a previous work, a PENELOPE Monte Carlo model of a Cyberknife system equipped with fixed collimator was developed and validated for in-field dose evaluation. The aim of this work is to extend it to evaluate peripheral doses and to determine the precision of the treatment planning system (TPS) Multiplan in evaluating the off-axis doses. The Cyberknife® head model was completed with surrounding components based on manufacturer drawings. The contribution of the different head parts on the out-of-field dose was studied. To model the attenuation and the modification of particle energy caused by components not modelled, the photon transport was modified in one of the added components. The model was iteratively adjusted to fit dose profiles measured with EBT3 films and an ionization chamber for several collimator sizes. Finally, dose profiles were calculated using the two Multiplan TPS algorithms and were compared to our simulations. The contributions to out-of-field dose were identified as scattered radiation from the phantom and head leakage and scatter originating at the secondary collimator level. Particle transport in the additional pieces was modified to model this radiation. The maximum differences between simulated and measured doses are of 20.4%. Regarding the detector responses away from axis, EBT3 films and the Farmer chamber give similar response (less than 20% difference). The TPS Monte Carlo algorithm underestimates the doses away from axis more importantly for the smaller field sizes (up to 98%). Besides, RayTracing simplifies peripheral dose to a constant value with no inclusion of particle transport. A Monte Carlo model of a Cyberknife system for the determination of out-of-field doses up to 14 cm off-axis was successfully developed and validated for different depths and field sizes in comparison with measurements. This study also confirms that TPS algorithms do not model peripheral dose properly.

Determination of the KQclinfclin,Qmsr fmsr correction factors for detectors used with an 800 MU/min CyberKnife(®) system equipped with fixed collimators and a study of detector response to small photon beams using a Monte Carlo method.

PURPOSE: In a previous work, output ratio (ORdet) measurements were performed for the 800 MU/min CyberKnife(®) at the Oscar Lambret Center (COL, France) using several commercially available detectors as well as using two passive dosimeters (EBT2 radiochromic film and micro-LiF TLD-700). The primary aim of the present work was to determine by Monte Carlo calculations the output factor in water (OFMC,w) and the [Formula: see text] correction factors. The secondary aim was to study the detector response in small beams using Monte Carlo simulation. METHODS: The LINAC head of the CyberKnife(®) was modeled using the PENELOPE Monte Carlo code system. The primary electron beam was modeled using a monoenergetic source with a radial gaussian distribution. The model was adjusted by comparisons between calculated and measured lateral profiles and tissue-phantom ratios obtained with the largest field. In addition, the PTW 60016 and 60017 diodes, PTW 60003 diamond, and micro-LiF were modeled. Output ratios with modeled detectors (ORMC,det) and OFMC,w were calculated and compared to measurements, in order to validate the model for smallest fields and to calculate [Formula: see text] correction factors, respectively. For the study of the influence of detector characteristics on their response in small beams; first, the impact of the atomic composition and the mass density of silicon, LiF, and diamond materials were investigated; second, the material, the volume averaging, and the coating effects of detecting material on the detector responses were estimated. Finally, the influence of the size of silicon chip on diode response was investigated. RESULTS: Looking at measurement ratios (uncorrected output factors) compared to the OFMC,w, the PTW 60016, 60017 and Sun Nuclear EDGE diodes systematically over-responded (about +6% for the 5 mm field), whereas the PTW 31014 Pinpoint chamber systematically under-responded (about -12% for the 5 mm field). ORdet measured with the SFD diode and PTW 60003 diamond detectors were in good agreement with OFMC,w except for the 5 mm field size (about -7.5% for the diamond and +3% for the SFD). A good agreement with OFMC,w was obtained with the EBT2 film and micro-LiF dosimeters (deviation less than 1.4% for all fields investigated). [Formula: see text] correction factors for several detectors used in this work have been calculated. The impact of atomic composition on the dosimetric response of detectors was found to be insignificant, unlike the mass density and size of the detecting material. CONCLUSIONS: The results obtained with the passive dosimeters showed that they can be used for small beam OF measurements without correction factors. The study of detector response showed that ORdet is depending on the mass density, the volume averaging, and the coating effects of the detecting material. Each effect was quantified for the PTW 60016 and 60017 diodes, the micro-LiF, and the PTW 60003 diamond detectors. None of the active detectors used in this work can be recommended as a reference for small field dosimetry, but an improved diode detector with a smaller silicon chip coated with tissue-equivalent material is anticipated (by simulation) to be a reliable small field dosimetric detector in a nonequilibrium field.

A new single crystal diamond dosimeter for small beam: comparison with different commercial active detectors.

Recent developments of new therapy techniques using small photon beams, such as stereotactic radiotherapy, require suitable detectors to determine the delivered dose with a high accuracy. The dosimeter has to be as close as possible to tissue equivalence and to exhibit a small detection volume compared to the size of the irradiation field, because of the lack of lateral electronic equilibrium in small beam. Characteristics of single crystal diamond (tissue equivalent material Z = 6, high density) make it an ideal candidate to fulfil most of small beam dosimetry requirements. A commercially available Element Six electronic grade synthetic diamond was used to develop a single crystal diamond dosimeter (SCDDo) with a small detection volume (0.165 mm(3)). Long term stability was studied by irradiating the SCDDo in a (60)Co beam over 14 h. A good stability (deviation less than ± 0.1%) was observed. Repeatability, dose linearity, dose rate dependence and energy dependence were studied in a 10 × 10 cm(2) beam produced by a Varian Clinac 2100 C linear accelerator. SCDDo lateral dose profile, depth dose curve and output factor (OF) measurements were performed for small photon beams with a micro multileaf collimator m3 (BrainLab) attached to the linac. This study is focused on the comparison of SCDDo measurements to those obtained with different commercially available active detectors: an unshielded silicon diode (PTW 60017), a shielded silicon diode (Sun Nuclear EDGE), a PinPoint ionization chamber (PTW 31014) and two natural diamond detectors (PTW 60003). SCDDo presents an excellent spatial resolution for dose profile measurements, due to its small detection volume. Low energy dependence (variation of 1.2% between 6 and 18 MV photon beam) and low dose rate dependence of the SCDDo (variation of 1% between 0.53 and 2.64 Gy min(-1)) are obtained, explaining the good agreement between the SCDDo and the efficient unshielded diode (PTW 60017) in depth dose curve measurements. For field sizes ranging from 0.6 × 0.6 to 10 × 10 cm(2), OFs obtained with the SCDDo are between the OFs measured with the PinPoint ionization chamber and the Sun Nuclear EDGE diode that are known to respectively underestimate and overestimate OF values in small beam, due to the large detection volume of the chamber and the non-water equivalence of both detectors.

Small fields output factors measurements and correction factors determination for several detectors for a CyberKnife® and linear accelerators equipped with microMLC and circular cones.

PURPOSE: The use of small photon fields is now an established practice in stereotactic radiosurgery and radiotherapy. However, due to a lack of lateral electron equilibrium and high dose gradients, it is difficult to accurately measure the dosimetric quantities required for the commissioning of such systems. Moreover, there is still no metrological dosimetric reference for this kind of beam today. In this context, the first objective of this work was to determine and to compare small fields output factors (OF) measured with different types of active detectors and passive dosimeters for three types of facilities: a CyberKnife(®) system, a dedicated medical linear accelerator (Novalis) equipped with m3 microMLC and circular cones, and an adaptive medical linear accelerator (Clinac 2100) equipped with an additional m3 microMLC. The second one was to determine the kQclin,Qmsr (fclin,fmsr) correction factors introduced in a recently proposed small field dosimetry formalism for different active detectors. METHODS: Small field sizes were defined either by microMLC down to 6 × 6 mm(2) or by circular cones down to 4 mm in diameter. OF measurements were performed with several commercially available active detectors dedicated to measurements in small fields (high resolution diodes: IBA SFD, Sun Nuclear EDGE, PTW 60016, PTW 60017; ionizing chambers: PTW 31014 PinPoint chamber, PTW 31018 microLion liquid chamber, and PTW 60003 natural diamond). Two types of passive dosimeters were used: LiF microcubes and EBT2 radiochromic films. RESULTS: Significant differences between the results obtained by several dosimetric systems were observed, particularly for the smallest field size for which the difference in the measured OF reaches more than 20%. For passive dosimeters, an excellent agreement was observed (better than 2%) between EBT2 and LiF microcubes for all OF measurements. Moreover, it has been shown that these passive dosimeters do not require correction factors and can then be used as reference dosimeters. Correction factors for the active detectors have then been determined from the mean experimental OF measured by the passive dosimeters. CONCLUSIONS: Four sets of correction factors needed to apply the new small field dosimetry formalism are provided for several active detectors. A protocol for small photon beams OF determination based on passive dosimeters measurements has been recently proposed to French radiotherapy treatment centers.

Numerical dosimetric reconstruction of a radiological accident in South America in April 2009.

A severe irradiation accident involving a victim occurred in April 2009 in South America. The victim has found a (192)Ir source fallen from a gammagraphy device and has put it in the left pocket of his pants. Very quickly, an erythema and a blister appeared on the left leg of the victim involving hospitalisation. Following the request of the IAEA assistance, the Ionizing Radiation Dosimetry Laboratory of IRSN was asked to perform a numerical dosimetric reconstruction. A personalised voxel phantom of the victim has been constructed thanks to the Simulation of External Source Accident with Medical images tool developed by the laboratory, and a calculation of the dose with the MCNPX computer code allowed to determine the boundary of the necrotic dose at 25 Gy. On the basis of these calculations, the physicians have performed exeresis of the necrotic region on the left leg on 4 May 2009. Associated with mesenchymal stem cell injection, the leg of the victim was healthy on December 2009.

Lessons from recent accidents in radiation therapy in France.

Many accidents in radiotherapy have been reported in France over the last years. This is due to the recent legal obligation to declare to the national safety authorities any significant incident relative to the use of ionising radiation including medical applications. The causes and consequences of the most serious events in radiotherapy are presented in this paper. Lessons can be learned from possible technical dysfunctions, from human errors or organisational weaknesses as to how such events can be prevented. The technical aspects are addressed here: in particular, dosimetric issues.

New approach to radiation burn treatment by dosimetry-guided surgery combined with autologous mesenchymal stem cell therapy.

The therapeutic management of severe radiation burns remains a challenging issue. Conventional surgical treatment (excision and skin autograft or rotation flap) often fails to prevent unpredictable and uncontrolled extension of the radiation necrotic process. We report here an innovative therapeutic strategy applied to the victim of a radiation accident (December 15, 2005) with an iridium gammagraphy radioactive source (192Ir, 3.3 TBq). The approach combined numerical dosimetry-guided surgery with cellular therapy using mesenchymal stem cells. A very severe buttock radiation burn (2000 Gy at the center of the skin surface lesion) of a 27-year-old Chilean victim was widely excised (10 cm in diameter) using a physical and anatomical dose reconstruction in order to better define the limit of the surgical excision in apparently healthy tissues. A secondary extension of the radiation necrosis led to a new excision of fibronecrotic tissues associated with a local cellular therapy using autologous expanded mesenchymal stem cells as a source of trophic factors to promote tissue regeneration. Bone marrow-derived mesenchymal stem cells were expanded according to a clinical-grade technique using closed culture devices and serum-free medium enriched in human platelet lysate. The clinical evolution (radiation pain and healing progression) was favorable and no recurrence of radiation inflammatory waves was observed during the 11 month patient's follow-up. This novel multidisciplinary therapeutic approach combining physical techniques, surgical procedures and cellular therapy with adult stem cells may be of clinical relevance for improving the medical management of severe localized irradiations. It may open new prospects in the field of radiotherapy complications.

Uncertainty analysis of the weighted equivalent lung dose per unit exposure to radon progeny in the home.

A parameter uncertainty analysis has been performed to derive the probability distribution of the weighted equivalent dose to lung for an adult (w(lung) H(lung)) per unit exposure to radon progeny in the home. The analysis was performed using the ICRP Publication 66 human respiratory tract model (HRTM) with tissue weighting factor for the lung, w(lung) = 0.12 and the radiation weighting factor for alpha particles, wR = 20. It is assumed that the HRTM is a realistic representation of the physical and biological processes, and that the parameter values are uncertain. The parameter probability distributions used in the analysis were based on a combination of experimental results and expert judgement from several prominent European scientists. The assignment of the probability distributions describing the uncertainty in the values of the assigned fractions (ABB, Abb, AAI) of the tissue weighting factor proved difficult in practice due to lack of quantitative data. Because of this several distributions were considered. The results of the analysis give a mean value of w(lung) H(lung) per unit exposure to radon progeny in the home of 15 mSv per working level month (WLM) for a population. For a given radon gas concentration, the mean value of w(lung) H(lung) per unit exposure is 13 mSv per 200 Bq.m(-3).y of 222Rn. Parameters characterising the distributions of w(lung) H(lung) per unit exposure are given. If the ICRP weighting factors are fixed at their default values (ABB, Abb, AAI = 0.333, 0.333, 0.333; w(lung) = 0.12; and wr = 20) then on the basis of this uncertainty analysis it is extremely unlikely (P approximately 0.0007) that a value of Hw/Pp for exposure in the home is as low as 4 mSv per WLM, the value determined with the epidemiological approach. Even when the uncertainties in the ABB, Abb, AAI, values are included then this probability is predicted to be between 0.01 to 0.08 depending upon the distribution assumed for describing the uncertainties in the ABB, Abb, AAI, values. Thus, it is concluded that the uncertainties in the HRTM parameters considered in this study cannot totally account for the discrepancy between the dosimetric and epidemiological approaches.

[Bacillary angiomatosis in an adult infected with HIV-1 at an early stage of immunodepression in Abidjan, Côte d'Ivoire]. / Angiomatose bacillaire chez un adulte infecté par le VIH-1 à un stade d'immunodépression peu avancé à Abidjan, Côte d'Ivoire.

Human immunodeficiency virus (HIV)-associated bacillary angiomatosis has rarely been described in Africa. We report here the first case in Côte d'Ivoire. Although in industrialised countries bacillary angiomatosis has been described in patients with low CD4 count, this episode occurred in the first year following HIV-seroconversion in an adult patient with more than 500 CD4 cells per cubic millimetre. Symptoms rapidly and totally disappeared under erythromycin treatment, although with a relapse two years after the end of the first episode. In Africa where people living with HIV often present chronic cutaneous lesions, bacillary angiomatosis may be under-diagnosed. Bacillary angiomatosis must be systematically considered in face of lesions similar to Kaposi's sarcoma. Improving knowledge on symptoms of bacillary angiomatosis in Africa should lead to better treatment and a better estimation of its true frequency which may be underestimated.

Laminin-alpha6beta1 integrin interaction enhances survival and proliferation and modulates steroidogenesis of ovine granulosa cells.

This study aimed to determine the physiological role of laminin (LN) and its receptor, alpha(6)beta(1) integrin, in controlling the functions of granulosa cells (GC) during follicular development in sheep ovary. Immunohistochemistry experiments showed the presence of increasing levels of LN (P<0.0001), and high levels of mature alpha(6)beta(1) integrin in GC layers of healthy antral follicles during the follicular and the preovulatory phases of the estrous cycle. In vitro, the addition of a function-blocking antibody raised against alpha(6) subunit (anti-alpha(6) IgG) to the medium of ovine GC cultured on LN impaired cell spreading (P<0.0001), decreased the proliferation rate (P<0.05) and increased the apoptosis rate (P<0.05). Furthermore, addition of anti-alpha(6) IgG enhanced estradiol (E2) secretion by GC in the presence or absence of follicle-stimulating hormone (FSH), luteinizing hormone or insulin-like growth factor-I in culture medium (P<0.0001), and inhibited progesterone (P4) secretion in basal conditions or in the presence of low (0.5 ng/ml) FSH concentrations only (P<0.0001). The anti-alpha(6) IgG effect was specific to an interaction of LN with alpha(6)beta(1) integrin since it was ineffective on GC cultured on heat-denatured LN, RGD (arginine-glycine-aspartic acid) peptides and non-coated substratum. Hence, this study established that alpha(6)beta(1) integrin 1) was expressed in GC of antral follicles, 2) mediated the actions of LN on survival, proliferation and steroidogenesis of GC, and 3) was able to dramatically modulate P4 and E2 secretion by GC in vitro. It is suggested that during the follicular and the preovulatory phases of the estrous cycle, the increasing levels of LN in GC of large antral follicles might support their final development to ovulation.

Skeletal muscle cell hypertrophy induced by inhibitors of metalloproteases; myostatin as a potential mediator.

Cell growth and differentiation are controlled in many tissues by paracrine factors, which often require proteolytic processing for activation. Metalloproteases of the metzincin family, such as matrix metalloproteases and ADAMs, recently have been shown to be involved in the shedding of growth factors, cytokines, and receptors. In the present study, we show that hydroxamate-based inhibitors of metalloproteases (HIMPs), such as TAPI and BB-3103, increase the fusion of C(2)C(12) myoblasts and provoke myotube hypertrophy. HIMPs did not seem to effect hypertrophy via proteins that have previously been shown to regulate muscle growth in vitro, such as insulin-like growth factor-I, calcineurin, and tumor necrosis factor-alpha. Instead, the proteolytic maturation of myostatin (growth differentiation factor-8) seemed to be reduced in C(2)C(12) cells treated with HIMPs, as suggested by the presence of nonprocessed myostatin precursor only in hypertrophic myotubes. Myostatin is a known negative regulator of skeletal muscle growth, belonging to the transforming growth factor-beta/bone morphogenetic protein superfamily. These results indicate that metalloproteases are involved in the regulation of skeletal muscle growth and differentiation, that the proteolytic maturation of myostatin in C(2)C(12) cells may be directly or indirectly linked to the activity of some unidentified HIMP-sensitive metalloproteases, and that the lack of myostatin processing on HIMP treatment may be a mediator of myotube hypertrophy in this in vitro model.

Extracellular matrix regulates ovine granulosa cell survival, proliferation and steroidogenesis: relationships between cell shape and function.

The extracellular matrix (ECM), constituting the follicular basal lamina and present also between follicular cells and in the follicular fluid, is believed to regulate granulosa cell (GC) function during follicular development. Ovine GCs isolated from small (1-3 mm in diameter) or large (4-7 mm in diameter) antral follicles were cultured on various pure ECM components (type I collagen, fibronectin, laminin), synthetic substrata enhancing (RGD peptides) or impairing (poly 2-hydroxyethylmethacrylate (poly-hema)) cell adhesion, or in the presence of heparin. The effects of these factors, used alone or in combination with IGF-I and/or FSH, were evaluated in terms of GC spread, survival, proliferation and steroidogenesis. When grown on type I collagen (CI) gel, poly-hema or heparin, GCs from both large and small follicles exhibited a round shape and a low proliferation rate. Compared with non-coated plastic substratum as a control, these ECM or synthetic compounds enhanced estradiol secretion and reduced progesterone secretion by large-follicle GCs. In contrast, GCs from both large and small follicles spread extensively on CI coating, fibronectin, laminin and RGD peptides. Fibronectin and laminin dramatically increased the proliferation rate and enhanced survival of GCs from both origins. Moreover, fibronectin, laminin and RGD peptides reduced estradiol secretion by large-follicle GCs. Unexpectedly, CI coating increased estradiol secretion and reduced progesterone secretion by large-follicle GCs, suggesting that type I collagen was able to maintain estradiol secretion independently of GC shape. Finally, GC responsiveness to IGF-I and FSH, in terms of proliferation and steroidogenesis, was generally maintained when cells were grown on ECM components, RGD peptides and in the presence of heparin. However, when large-follicle GCs were grown as non-adherent clusters (as observed on poly-hema) basal and IGF-I- and/or FSH-stimulated progesterone secretions were totally abolished. Overall, this study shows that GC shape, survival, proliferation and steroidogenesis can be modulated in vitro by pure ECM components in a specific and coordinated manner. It is suggested that, in vivo, fibronectin and laminin would sustain follicular development by enhancing the survival and proliferation of GCs, whereas type I collagen might participate in the maintenance of estradiol secretion in large antral follicles.

A meta-analysis of the effectiveness of cell salvage to minimize perioperative allogeneic blood transfusion in cardiac and orthopedic surgery. International Study of Perioperative Transfusion (ISPOT) Investigators.

UNLABELLED: Concern about risks of allogeneic transfusion has led to an interest in methods for decreasing perioperative transfusion. To determine whether cell salvage reduces patient exposure to allogeneic blood, we performed meta-analyses of randomized trials, evaluating the effectiveness and safety of cell salvage in cardiac or orthopedic elective surgery. The primary outcome was the proportion of patients who received at least one perioperative allogeneic red cell transfusion. Twenty-seven studies were included in the meta-analyses. Cell salvage devices that do not wash salvaged blood were marginally effective in cardiac surgery patients when used postoperatively (relative risk [RR] = 0.85, 95% confidence interval [CI] = 0.79-0.92). Devices that wash or do not wash salvaged blood considerably decreased the proportion of orthopedic surgery patients who received allogeneic transfusion (RR = 0.39, 95% CI = 0.30-0.51 and RR = 0.35, 95% CI 0.26-0.46, respectively). No studies of cell savers that wash salvaged blood during cardiac surgery were included. Cell salvage did not appear to increase the frequency of adverse events. We conclude that cell salvage in orthopedic surgery decreases the risk of patients' exposure to allogeneic blood transfusion perioperatively. Postoperative cell salvage in cardiac surgery, with devices that do not wash the salvaged blood, is only marginally effective. IMPLICATIONS: This meta-analysis of all published randomized trials provides the best current estimate of the effectiveness of cell salvage and is useful in guiding clinical practice. We conclude that cell salvage in orthopedic surgery decreases the proportion of patients requiring allogeneic blood transfusion perioperatively, but postoperative cell salvage is only marginally effective in cardiac surgery.

[Cost-benefit analysis of screening strategies by human immunodeficiency virus in French blood donors]. / Analyse coût-efficacité des stratégies de dépistage de l'infection par le virus de l'immunodéficience humaine sur les dons de sang en France.

BACKGROUND: The residual risk of human immunodeficiency virus (HIV) infection from screened blood transfusion was estimated to be 1.7/10(6) between 1993 and 1995 in France. To orient blood safety policies, we have evaluated what would be, from the perspective of blood banks, the best screening strategy in terms of gain in effectiveness and added costs. METHODS: A cost-effectiveness analysis compared 20 HIV-testing protocols using (1) available data for performances of the current screening tests; and, (2) national insurance estimates for the cost of tests. Results were expressed as the number of false negative donations that would be avoided and the cost by avoided false negative donation. RESULTS: For 3 million donated blood units a year and a prevalence of 24 per million, there would be 72 infectious donated blood units, 70.56 of which would be detected by the current screening strategy. The number of additional donated blood infections avoided in all other strategies would be low (between 0.25 and 1.28) with a very high cost (280 million French francs per added false negative avoided or more). CONCLUSION: A change in screening strategies for blood donations in France is not currently justified. If such a change was to be done, adding p24 antigen detection to the current screening strategy would be one of the worst solutions.

Technologies to minimize blood transfusion in cardiac and orthopedic surgery. Results of a practice variation survey in nine countries. International Study of Peri-operative Transfusion (ISPOT) Investigators.

OBJECTIVES: Due to the discovery in the 1980s that blood transfusion can transmit HIV, there has been increased interest in technologies that reduce the amount of allogeneic blood used during and after surgery. These technologies include drugs (aprotinin, tranexamic acid, epsilon-aminocaproic acid, erythropoietin), devices (cell salvage), and techniques (acute hemodilution, predeposited autologous donation). The purpose of this study was to ascertain the degree of practice variation, if any, that exists for eight technologies in nine countries in orthopedic and cardiac surgery. METHODS: In each country, either all hospitals or a random sample of hospitals with medical/surgical beds were surveyed between 1995 and 1997. Two instruments were used. The first instrument was a postcard that asked recipients whether the technologies were currently being used in their hospital for orthopedic and/or cardiac surgery to reduce perioperative allogeneic transfusion. The second questionnaire elicited information regarding the degree of use both in qualitative and quantitative terms. Data were collected, entered, and analyzed in each country, with summary results submitted to the Canadian coordinating center on a standardized data collection form. RESULTS: Pharmaceuticals were generally used in a much smaller proportion of hospitals in orthopedic than in cardiac surgery. Aprotinin and tranexamic acid were the drugs most frequently used in cardiac surgery. Nonpharmacological technologies were used to a greater degree than drugs in orthopedic surgery, although there was wide variation among technologies and countries. Acute hemodilution and cell salvage were used in a greater proportion of hospitals for cardiac surgery than orthopedic surgery. CONCLUSIONS: The results of this survey indicate that there is considerable practice variation in the use of technologies to minimize exposure to perioperative allogeneic transfusion within and between countries.

Chronology of events accompanying follicular atresia in hypophysectomized ewes. Changes in levels of steroidogenic enzymes, connexin 43, insulin-like growth factor II/mannose 6 phosphate receptor, extracellular matrix components, and matrix metalloproteinases.

The chronology of changes in levels of some proteins known to be altered during atresia of ovarian follicles was studied in ewes hypophysectomized at the end of the follicular phase of the estrous cycle. This study was performed by quantitative immunohistochemistry and zymography on large antral follicles (diameter > 3.5 mm) normally destined to ovulate, recovered 24, 36, or 72 h after pituitary ablation. The process of atresia was followed by comparing healthy follicles from intact ewes, with early atretic follicles recovered 24 h after hypophysectomy, clearly atretic follicles recovered 36 h after hypophysectomy, and late atretic follicles recovered 72 h after hypophysectomy. The earliest events of atresia induced by hypophysectomy were detected in theca and consisted of a strong decrease in levels of fibronectin (p < 0.001) and insulin-like growth factor II/mannose 6 phosphate (IGF-II/M6P) receptors (p < 0.05), occurring within the first 24 h following pituitary ablation. In intact animals, similar changes were observed in theca of early atretic follicles, suggesting that these changes may be important events involved in the onset of follicular atresia. In parallel, intrafollicular levels of matrix metalloproteinase-2 and -9 were shown to increase as early as 24 h after hypophysectomy, and a further increase was observed until 72 h after hypophysectomy. These early events were followed by the disappearance of P450 aromatase in granulosa cells 36 h after hypophysectomy (p < 0.05), and a progressive decrease in levels of P450 17alpha-hydroxylase in the theca interna and of gap junction protein connexin-43 in granulosa cells; these markers were still detectable in late atretic follicles 72 h after hypophysectomy. The increase in levels of fibronectin, type IV collagen, laminin, and IGF-II/M6P receptors within granulosa cell layers (p < 0.05) was significant only in late atretic stages, suggesting that these changes may be consequences rather than causes of atresia.

[Bazex paraneoplastic acrokeratosis. Treatment with acitretin]. / Acrokératose paranéoplasique de Bazex. Traitement par acitrétine.

INTRODUCTION: Bazex paraneoplastic acrokeratosis remains a therapeutic challenge when the cancer cannot be treated. We report the third patient in which a complete clearance of the lesions was obtained with oral acitretine. CASE REPORT: A 67 year-old patient had Bazex paraneoplastic acrokeratosis. Despite a complete cancer screening, no cancer was found. A cervical lymph node metastasis was treated with surgery and radiotherapy. Because the cutaneous lesions got worse after this treatment, the patient received oral acitretine. A complete clearing of the lesions was observed within 2 months. DISCUSSION: Only 2 similar cases have been reported to our knowledge. The comparison with these 2 cases supports the efficacy of oral retinoids in this disease. The search and the treatment of the cancer remain compulsory. CONCLUSION: In some definite cases, oral retinoids can be proposed as a treatment of cutaneous lesions in Bazex paraneoplastic acrokeratosis.

Comparative analysis of neutral endopeptidase (NEP) and villin gene expression during mouse embryogenesis and enterocyte maturation.

Neutral endopeptidase (Endopeptidase 24.11; NEP; neprilysin), an integral membrane protein, and villin, a major microvillar cytoskeletal actin-binding protein, are both typically associated with brush border epithelia. In this study, cRNA probes were hybridized in situ to investigate the expression of NEP and villin genes in embryo and adult mouse enterocytes. During development, villin mRNAs were easily detected in the immature digestive tract well before establishment of the brush border. In 17-day-old embryos, a transient elevation of villin mRNA occurred just prior to a dramatic increase in microvilli length and density. NEP only appeared by day 17 as the embryonic gut began to become functional. It therefore appears that the onset of transcription of specialized cytoskeletal proteins from the brush border preceded that of intrinsic membrane-bound enzyme from microvilli. In the adult intestinal fold, both mRNAs were expressed along the whole length of the villus with maximal expression at its base. In contrast, both proteins were uniformly expressed along the whole crypt-villus axis. Quantitative analysis revealed an asymmetric intracellular distribution of both mRNAs that were differentially polarized in the apical cytoplasm of enterocytes.

Water handling in Caco-2 cells: effects of acidification of the medium.

Caco-2 cells were cultured on permeable supports. At confluence the minute-by-minute net water movement (Jw) was automatically recorded. Simultaneously, unidirectional [14C]mannitol, 22Na+, and/or 36Cl- fluxes and transepithelial resistances were measured. The water and mannitol permeabilities went progressively down between 9 and 16 days after seeding and then stabilized. In this last condition the hydrostatic permeability coefficient (Phydr) was 2.67 +/- 0.31 cm s-1 while the osmotic permeability coefficient (Posm) was 0.0017 +/- 0.0004 cm s-1. Phydr but not Posm was dependent on the temperature and on the presence of Na+ in the medium. A net secretory Jw was observed 16 days after seeding, in the absence of any osmotic, hydrostatic or chemical gradient. This secretory Jw was associated with net Cl- (1.43 +/- 0.43 muequiv h-1 cm-2) and Na+ (1.05 +/- 0.35 muequiv h-1 cm-2) secretions. Amiloride reduced, in open-circuit conditions, both Na+ and Cl- apical to basal fluxes, thus enhancing the net Na+ and Cl- exit. Acidification of the medium (pH 6.2) reversibly increased water and mannitol permeabilities in 10-day-old cultures. In 16-day-old cultures the same shift in medium pH did not change mannitol permeability, while stimulating water secretion. These results, obtained in the absence of supracellular structures (villae, crypts) and subepithelial components (muscular, vascular and conjunctive tissues) indicate that paracellular and transport-associated water pathways are sensitive to changes in the pH of the medium in Caco-2 cell layers.