The CONJUDOR pipeline for multiplexed knockdown of gene pairs identifies RBBP-5 as a germ cell reprogramming barrier in C. elegans.

Multiple gene activities control complex biological processes such as cell fate specification during development and cellular reprogramming. Investigating the manifold gene functions in biological systems requires also simultaneous depletion of two or more gene activities. RNA interference-mediated knockdown (RNAi) is commonly used in Caenorhabditis elegans to assess essential genes, which otherwise lead to lethality or developmental arrest upon full knockout. RNAi application is straightforward by feeding worms with RNAi plasmid-containing bacteria. However, the general approach of mixing bacterial RNAi clones to deplete two genes simultaneously often yields poor results. To address this issue, we developed a bacterial conjugation-mediated double RNAi technique 'CONJUDOR'. It allows combining RNAi bacteria for robust double RNAi with high-throughput. To demonstrate the power of CONJUDOR for large scale double RNAi screens we conjugated RNAi against the histone chaperone gene lin-53 with more than 700 other chromatin factor genes. Thereby, we identified the Set1/MLL methyltransferase complex member RBBP-5 as a novel germ cell reprogramming barrier. Our findings demonstrate that CONJUDOR increases efficiency and versatility of RNAi screens to examine interconnected biological processes in C. elegans with high-throughput.

A new laparoscopic method of bowel radio-protection before pelvic chemoradiation of locally advanced cervix cancers.

BACKGROUND: Chemoradiation therapy (CRT) has become the mainstay of locally advanced cervical carcinomas (LACC). However, the price to pay is a significant rate of both early and late colo-rectal toxicities, which may impact on survivors' quality of life. To reduce the incidence of such complications, we suggest a simple technique of pelvic radioprotection. MATERIALS AND METHODS: An omental flap is created which is placed to fill the Douglas pouch to both increase the space between rectum and uterine cervix and prevent small bowel to fall in and to be exposed to radiation. In addition, a long sigmoid loop is retracted and fixed in the left paracolic gutter to prevent its irradiation as well. RESULTS: From May 2011 to May 2012, 51 successive LACC patients were offered this procedure in addition of a laparoscopic staging. All but 2 with too small an omentum benefitted from omentoplasty, while sigmoidopexy was performed in all but one patient with a long and free sigmoid loop. No immediate adverse effect was observed. The volume of retro-uterine omental flap averaged 7.17 ± 3.79 cm(3). Sequential measurements of the utero-rectal space throughout CRT duration showed a real and durable increase in the distance between these organs, resulting in a drop in the dose of irradiation to recto-sigmoid. With 10 ± 4.5-month median follow-up, we did not observe any rectal or small bowel early or late adverse effects of CRT. CONCLUSIONS: Although this series is preliminary, this simple procedure, feasible by laparoscopy (or laparotomy), seems effective to prevent recto-sigmoid as well as small bowel from radio-induced complications due to pelvic CRT.

Multi-center planning study of radiosurgery for intracranial metastases through Automation (MC-PRIMA) by crowdsourcing prior web-based plan challenge study.

BACKGROUND: Planning radiosurgery to multiple intracranial metastases is complex and shows large variability in dosimetric quality among planners and treatment planning systems (TPS). This project aimed to determine whether autoplanning using the Muliple Brain Mets (AutoMBM) software can improve plan quality and reduce inter-planner variability by crowdsourcing results from prior international planning study. METHODS: Twenty-four institutions autoplanned with AutoMBM on a five metastases case from a prior international planning competition from which population statistics (means and variances) of 23 dosimetric metrics and resulting composite plan score (maximum score = 150) of other TPS (Eclipse, Monaco, RayStation, iPlan, GammaPlan, MultiPlan) were crowdsourced. Plan results of AutoMBM and each of the other TPS were compared using two sample t-tests for means and Levene's tests for variances. Plan quality of AutoMBM was correlated with the planner' experience and compared between academic and non-academic centers. RESULTS: AutoMBM produced plans with comparable composite plan score to GammaPlan, MultiPlan, Eclipse and iPlan (127.6 vs. 131.7 vs. 127.3 vs. 127.3 and 126.7; all p > 0.05) and superior to Monaco and RayStation (118.3 and 108.6; both p < 0.05). Inter-planner variability of overall plan quality was lowest for AutoMBM among all TPS (all p < 0.05). AutoMBM's plan quality did not differ between academic and non-academic centers and uncorrelated with planning experience (all p > 0.05). CONCLUSIONS: By plan crowdsourcing prior international plan challenge, AutoMBM produces high and consistent plan quality independent of the planning experience and the institution that is crucial to addressing the technical bottleneck of SRS to intracranial metastases.

Brachmann-de Lange syndrome: pre- and postnatal findings.

Brachmann-de Lange syndrome (BDLS) is a well-delineated and relatively common syndrome. However, prenatal diagnosis has never been reported, even if in some cases ultrasonography demonstrated one or more manifestations of the syndrome. We report on 3 cases: in the first 2 cases, prenatal ultrasonography demonstrated some signs of the condition. The third represents, to our knowledge, the first prenatal diagnosis of BDLS. We also present a review of the literature concerning pre- and postnatal findings in this syndrome.

Computed tomographic evaluation of silicosis and coal workers' pneumoconiosis.

The introduction of computed tomography (CT) has modified the radiologic approach to silicosis and coal worker's pneumoconiosis considerably, allowing earlier recognition and more precise characterization of pulmonary abnormalities than chest radiography. An optimal approach to CT recognition requires an understanding of the main physiologic and pathologic features that are known to occur in both pneumoconioses. This report focuses on the most common CT features of simple and complicated forms of silicosis and coal worker's pneumoconiosis as well as on the optimal scanning protocol. The respective roles of chest radiographs and CT scans in the recognition of pneumoconiosis and follow-up of exposed workers are discussed.

Non-Hodgkin's lymphoma and Sjögren's syndrome. An immunopathological study of 113 patients.

One hundred and thirteen patients with untreated non-Hodgkin's lymphoma were evaluated for Sjögren's syndrome (SS) using clinical and biological parameters, and a minor salivary gland biopsy. Concomitant immunophenotyping for T and B cells, and kappa and lambda chains was performed on both minor salivary gland and lymphomatous tissues. Patients with a positive focus score on examination of their minor salivary gland biopsy had a second biopsy, when possible, at the end of induction therapy. Fourteen of the 113 patients had SS according to the Greek criteria, and 4 of these 14 patients had an identical monotypic infiltrate in both their minor salivary gland and lymphomatous tissues. In all four cases this infiltrate disappeared from both locations when the lymphoma was in complete remission (CR). The 10 other patients with criteria for SS had a mixed infiltrate which persisted unchanged when the lymphoma was in CR. Among the 99 patients without SS, 12 had a positive focus score and an identical monotypic infiltrate in both their minor salivary gland and lymphomatous tissue. This infiltrate disappeared from both locations when the lymphoma was in CR. Among the remaining 87 patients, 82 had no minor salivary gland infiltrate, and 5 had a positive focus score with mixed lymphocytic infiltrate. Long term follow up and a large cooperative study are needed to better understand the immunopathologic lesions of patients of this latter type.

Pulmonary hypertension of the newborn and urogenital anomalies in two male siblings: a new family with misalignment of pulmonary vessels.

The association of pelvi-ureteric junction obstruction (PUJO) and rapidly fatal persistent pulmonary hypertension of the newborn (PPHN) has been observed in two male siblings. PUJO was also observed in a maternal uncle, whose daughter suffered from vesico-ureteral reflux (VUR). In both patients, histopathologic study of the lungs showed misalignment of pulmonary veins (MAPV), which is a rare autosomal recessive condition leading to severe PPHN and death within the neonatal period. It has occasionally been described associated with PUJO. The authors point out that: i: MAPV has to be carefully searched in case of PPHN; ii: PUJO could be an important finding associated with MAPV, and the only prenatal indication of this lethal condition.

Evaluation of the peripheral dose to uterus in breast carcinoma radiotherapy.

The absorbed dose outside of the direct fields of radiotherapy treatment (or peripheral dose, PD) is responsible for radiation exposure of the fetus in pregnant women. Because the radiological protection of the unborn child is of particular concern in the early period of the pregnancy, the aim of this study is to estimate the PD in order to assess the absorbed dose in the uterus in a pregnant patient irradiated for breast carcinoma therapy. The treatment was simulated on an Alderson-Rando anthropomorphic phantom, and the radiation dose to the fetus was measured using an ionisation chamber and thermoluminescence dosemeters. Two similar treatments plans with and without wedges were delivered, using a 6 MV photon beam with two isocentric opposite tangential fields with a total dose of 50 Gy, in accordance with common established procedures. Average field parameters for more than 300 patients were studied. Measurements showed the fetal dose to be slightly lower than 50 mGy, a level at which the risk to the fetus is uncertain, although several authors consider this value as the dose threshold for deterministic effects. The planning system (PS) underestimated PD values and no significant influence was found with the use of wedge filters.

[Human papillomavirus detection by the hybrid capture II assay with the liquid cell preservation solution Easyfix]. / Détection des HPV oncogènes par recueil sur milieu Easyfix.

The aim of this study is to validate the liquid cell preservation solution Easyfix for DNA detection of the HPV oncogene using the Hybrid Capture II method. 256 specimens were selected for the cytological study, possible biopsy and HPV oncogene search with the Easyfix fixative fluid and the Cervical Sampler transport medium. The results obtained with both mediums are comparable regardless of the cytological type. The relevance of a cytological study combined with the HPV search is stressed. To conclude, it is possible to put forward that the liquid cell preservation solution Easyfix can be used to detect the HPV oncogene using the Hybrid Capture II method.

[Erdheim-Chester disease. Clinico-pathologic study of two cases]. / Maladie d'Erdheim-Chester. Etude anatomo-clinique à propos de deux cas.

Erdheim-Chester disease is a rare visceral xanthogranulomatosis characterized by bilateral, symmetrical sclerosis of the metaphyseal regions of long bones and infiltration of foamy, lipid-laden histiocytes. Clinically, it ranges from an asymptomatic, focal process to a fatal, systemic disease. We report two new cases, different in their presentation and extension.

[Histological diagnosis after Pap smears detecting cancer or high-grade lesion of the cervix]. / Correspondance histologique des frottis cervico--utérins détectant un cancer ou une lésion de haut grade.

OBJECTIVE: The aim of the study is to analyze the histological-cytological correlations for Pap smears having detected cancer or high-grade squamous intra-epithelial lesion (HSIL) of the cervix. PATIENTS AND METHOD: The study about 311 women is retrospective. The average age is 36.4 years. Group 1 (histological diagnosis of high-grade or invasive lesion) includes 244 women (77.5%). Group 2 (histological diagnosis other than high-grade or invasive lesion) includes 37 women (11.9%) with a presumed diagnosis of HSIL. Group 3 (absence of histological follow-up) includes 30 women (9.6%) with a presumed diagnosis of HSIL. RESULTS: In group 1, the presumed cytological diagnosis is HSIL in 229 cases, squamous carcinoma in 11 cases and adenocarcinoma in two cases. In this group, the average delay between the Pap smear and the first histology is equal to two months. It is longer than 6 months in seven cases. The diagnosis of cancer or high-grade lesion is confirmed histologically on a first biopsy of the cervix in 196 cases, a second or a third biopsy in 10 cases, an endocervical curettage in six cases and a surgical specimen in 32 cases. In the group 2, the histological diagnosis is normal-benign in 14 cases (presumed cytological false positives) and condyloma-CIN 1 in 23 cases (presumed overevaluations). DISCUSSION AND CONCLUSION: Results highlight benefits of interactive exchanges between clinicians and pathologists, and the necessity of review of discordant cases by several pathologists in due time, with written comments and coding of the conclusions of the review. Histological follow-up is late or not done in some women.

[Report of a case of T-cell peripheral lymphoblastic lymphoma in an HIV patient]. / Presentación de un caso de linfoma linfoblástico periférico de células T en paciente VIH.

T-cell lymphomas in human immunodeficiency virus infections are rare, first case have being described in 1987, by Presant. Our purpose is to report the first T-cell Lymphoma case without epidermotropism in an HIV patient in Extremadura, and pioneer in Spain. Clinic extensive and histopathologic studies of cutaneous lesions were realized, including monoclonal antibodies tests. Peculiar clinical features were small bowel disease (MALT), gingiva, pericardium and skin involvement, with spontaneous resolution of skin nodules. Polychemotherapy with cyclophosphamide, doxorubicin, vincristine and prednisone was not effective, causing serious myelotoxicity. We outline the rarity of T-cell Lymphomas, the predominance of T4 phenotype, its relation with Epstein-Barr virus, the increase in 6-interleukin production, and the prognostic value of these factors, in correlation with advanced clinical status and unfavorable outcome.

[Intraoperative molecular assessment of sentinel nodes in the breast cancer using the Gene Search BLN Assay technique: our experience about 126 patients]. / Évaluation de l'analyse peropératoire des ganglions "sentinelle" par biologie moléculaire (technique Gene Search BLN Assay) dans le cancer du sein: à propos de 126 patientes.

INTRODUCTION: Intraoperative molecular assay Gene Search BLN Assay (BLN) detects sentinel lymph node (SLN) metastasis in breast cancer. Our objective was to compare BLN to the definitive conventional histologic methods and to experiment the management of BLN in routine. MATERIAL AND METHODS: Each SLN was cut into alternate slabs. Half slabs were analysed with the intraoperative BLN molecular method, and the other slabs with the definitive histologic method. RESULTS: Two hundred and thirty four SLN have been analysed (124 patients). Thirty-five SLN had metastasis for 29 patients (23.4%). BLN correctly identified 28 patients. Two cases of discordance between BLN and standard method were found, probably explained by a sample bias. The sensibility of BLN is 96.4%, the sensitivity is 99%, the predictive positive value is 96.4%, the predictive negative value is 99% and the concordance is 98.4%. The surgery time increases and there is a need to adapt the theatre organization accordingly. CONCLUSION: The Gene Search BLN Assay gives a great interest for the patient, the surgeon and the pathologist because it increases the quality of the intraoperative analysis by comparison with the intraoperative conventional histology.

Fluorescence of experimental endometriosis in rabbits, using tamoxifen-eosin association.

A major problem with endometriosis is the detection of microscopic and atypical lesions. An incomplete surgical eradication may lead to recurrence of the disease. This study aimed to investigate the diagnostic improvement of endometriosis by the use of tamoxifen-eosin induced fluorescence. The experimental study was performed on surgically induced endometriosis in the rabbit. Endometriosis was induced by grafting endometrium onto the broad ligament in 10 rabbits. After 5 weeks, the fluorescence of excised endometriosis was studied after systemic injection of tamoxifen and local application of eosin. Healthy peritoneal samples served as controls. The fluorescence of endometriotic foci was also compared with (n = 5) or without (n = 5) tamoxifen. Fluorescence excitation was carried out using a 150 W filtered lamp connected to an optical fibre. Fluorescence emission was measured using an optical fibre connected to a spectrofluorometer. Spectral analysis showed a specific fluorescence of endometriosis 72 h after systemic injection of tamoxifen and eosin application. This result is explained by binding to oestrogen receptors of tamoxifen which was protonized to form an ionic pair with eosin. Histological study of samples from the graft of endometrial tissue showed that experimental endometriosis had developed in eight out of the 10 rabbits. However, the fluorescence was not significantly different among the 10 rabbits. This observation was in accordance with previous studies in which endometriosis was confirmed by routine histological techniques or electron microscopy in 70-80% of cases. Consequently, the fluorescence of the two samples which did not present histological evidence of endometriosis indicates the presence of microscopic endometriotic foci.(ABSTRACT TRUNCATED AT 250 WORDS)

[Why we prefer the thin layer technique to conventional Pap smears. A double-blind study of 473 specimens]. / Pourquoi nous préférons la technique en couche mince aux frottis cervico-utérins conventionnels. Etude en double aveugle de 473 prélèvements.

The study evaluated the feasibility of a thinlayer technique on a routine basis for cervical smears and compared 473 ThinPrep preparations to the matched conventional Papsmears. The interpretation was double-blind and performed according to the Bethesda system. A consensus was established in discordant cases. The technique was easily mastered by gynecologists and technicians. Main advantages of the thinlayer technique were: a low number (1%) of unsatisfactory samples; a constant quality; homogeneity of cell distribution; the disappearance of problems of interpretation due to fixation or smear artifacts, red cells, polymorphs; a more precise interpretation, a firmer diagnosis; less visual fatigue; a shorter time of interpretation; the possibility of preparing more slides and performing special techniques on the same specimen. Main disadvantages were a higher cost and a necessary period of learning for cytologists.

Effectiveness and safety of prehospital urapidil for hypertensive emergencies.

The objective was to evaluate both the effectiveness and safety of urapidil in management of hypertensive emergencies (HE) in the prehospital setting. This was an open, prospective study for 6 months. We evaluated systolic (SBP), diastolic (DBP), and mean blood pressure (MBP), and heart rate (HR) with continuous noninvasive hemodynamic monitoring in 16 consecutive cases of HE. We used urapidil at different doses (25 to 100 mg) at 5 minute intervals, according to the blood pressure response. The basal DBP was 127 +/- 16 mmHg. Urapidil was effective in 15 patients. ANOVA test showed a significant drop out in DBP (P <.0001) and HR (P <.004). The highest decrease was obtained in the first 10 minutes. The decrease in DBP and HR values were significant at 5 minutes versus basal (P <.05) and at 10 versus 5 minutes (P <.01). All adverse effects had little relevance. Urapidil is effective and safe in management of HE when used by a medical team in the prehospital setting.