The Middle East and Eastern Europe rabies Expert Bureau (MEEREB) third meeting: Lyon-France (7-8 April, 2015).

MEEREB is an inter-regional network of countries from North Africa, Europe, the Middle East and Central Asia that work together with the aim of improving rabies control and prevention at local, regional and global level. MEEREB members met for the third time in 2015 in France (Lyon) to review the current rabies situation within the network and to discuss the way forward the prospect of a One Health approach against rabies. Dogs were the main vector of transmission in all MEEREB countries except for Croatia and Serbia where foxes represented the primary source. The number of rabies animal cases reported in 2014 varied substantially between countries with Ukraine reporting the highest number of animal cases. Human cases still occur in North Africa and all Middle East and Eurasian countries while no cases of human rabies were reported in Croatia, Serbia and Romania, although cases of rabies were identified in both dogs and foxes in 2014. Participants concluded that MEEREB can act as a think-tank where countries can share data, information, experiences and best practices to jointly address challenges in rabies control and prevention. They called for elimination of dog-transmitted rabies through vaccine and rabies immunoglobulin stockpiles and implementation of a One Health approach to achieve rabies's eradication.

[Field assessment of the new rapid diagnostic test Ebola eZYSCREEN®]. / Validation sur le terrain du nouveau test de diagnostic rapide Ebola eZYSCREEN®.

During the Ebola virus disease outbreak in West Africa in 2014, the World Health Organization has pointed out the need for rapid diagnostic tests (RDT) affordable, sensitive, specific, user-friendly, rapid, equipment-free, and deliverable. The rapid diagnostic test (Lateral Flow Assay) Ebola eZYSCREEN® was developed in this emergency frame using monoclonal antibodies against the envelope glycoprotein of the virus. Two distinct versions have been industrialized, one for whole-blood samples and the other for serum/plasma samples. Both versions have an analytical detection limit of 105 pfu/ml, the stability is at least 393 days at 30°C and 120 days at 45°C. The nonretrospective and independent validation study was carried out in the course of the outbreak in Conakry and at the Ebola Treatment Center of Coyah (Guinea) on 144 patients. In this study, the RDT showed a sensitivity of 65.3% and a specificity of 98.9% on whole blood, a sensitivity of 74.5% and a specificity of 100% on serum. Results from the whole-blood version must be analyzed with caution because of the delay between the blood collection and the completion of the tests, which was out of specification (3 days on average instead of 2 h). In contrast to laboratory tests, this easy to use field test does not require sophisticated instrumentation or even electricity and can contribute to the diagnostic chain of Ebola virus disease taking into account its benefits, high stability, and specificity but also its limit of sensitivity compared to laboratory techniques RT-qPCR (Real-Time reverse transcription Polymerase Chain Reaction), which remain the reference for the diagnosis of Ebola. The RDT Ebola eZYSCREEN® was granted EC IVD (IVD = In Vitro Diagnostic) marking.

Methods for assessing biochemical oxygen demand (BOD): a review.

The Biochemical Oxygen Demand (BOD) is one of the most widely used criteria for water quality assessment. It provides information about the ready biodegradable fraction of the organic load in water. However, this analytical method is time-consuming (generally 5 days, BOD5), and the results may vary according to the laboratory (20%), primarily due to fluctuations in the microbial diversity of the inoculum used. Work performed during the two last decades has resulted in several technologies that are less time-consuming and more reliable. This review is devoted to the analysis of the technical features of the principal methods described in the literature in order to compare their performances (measuring window, reliability, robustness) and to identify the pros and the cons of each method.

[Psychological adjustment of stereotactic breast biopsy instrumentation procedures: about 73 cases]. / Le vécu psychologique de la procédure de prélèvements percutanés du sein. Etude quasi prospective à propos de 73 cas.

OBJECTIVE: The aim of the study was to investigate patients' perceptions after stereotactic breast biopsy instrumentation, after both procedure and results. PATIENTS AND METHOD: From 1 March 2002 to 31 March 2003, a questionnaire (Likert response type) on stress was given to 73 patients who had breast biopsies procedures, the first time at the end of the procedure (T1) and then after the histological diagnosis (T2). RESULTS: The questionnaire was validated through analysis of principal component with Varimax rotation. Three factors were identified: procedure, quality of life, information and perception after biopsy. Responses were analysed with Chi-square. Two groups of women were identified, the first group (G1) corresponding to patients with a benign diagnosis (N=32) and the second group (G2) to patients with malignant diagnosis (N=32). Only the "procedure" factor was different at T1 and T2 (P=0.022). Compression was found to be painful: 11% at T1 versus 21% at T2. Women were disturbed by local anesthesia: 26% at T1 versus 21% at T2. Biopsy was painful: 6% at T1 versus 13% at T2. Examination was too long: 24% at T1 versus 35% at T2. The procedure was discomfortable: 52% at T1 versus 54% at T2. Information satisfied patients in 90% cases. There was no statistically significant difference according to procedures and histological disease (P=0.357). DISCUSSION AND CONCLUSION: Information and medical empathy conditioned patients' perceptions. Patients tolerated the procedures well.

[Study on the management of patients following detection of a "probably benign" mammographic abnormality (ACR BI-RADS category 3): a review of 176 radiologists]. / Une étude exploratoire des pratiques et des comportements radiologiques devant la découverte d'une anomalie "probablement bénigne" sur la mammographie: à propos de 176 radiologues.

OBJECTIVES: To evaluate mid-term management of patients with category 3 findings (probably benign, ACR BI-RADS) at mammography three months after a CME course on breast cancer. METHODS: A questionnaire (33 items) was sent to 529 radiologists, three months after the CME course (May-December 2002). The answers were analysed using the chi-square test. RESULTS: A total of 176 radiologists completed the questionnaire; 63 were part of an organized screening mammography program; 162 used the BI-RADS classification. In the presence of a class 3 finding, 158 radiologists recommended short term follow-up, 28 recommended biopsy, 9 upgraded the level of abnormality, 116 consulted another radiologist, 42 consulted an expert mammographer, 36 submitted the case to a multidisciplinary committee and 12 asked for an overread. Management with short term interval follow-up was worrisome to 91 radiologists for fear of unfavorable outcome, and to 47 radiologists because of possible litigation. Twenty-six radiologists had difficulty explaining to patients the need for short term follow-up. The median score for perceived stress by the radiologists was 3 (0=low and 10=high). CONCLUSION: The practices of this sample of French radiologists are consistent with the recommendations presented during the CME course and by the ANAES. They may have some difficulty in communicating with patients the need for follow-up. The ethics of radiology are based on "primum non nocere".

[Primary breast sarcoma. A retrospective study of 42 patients treated at the Bergonié Institute during a 32-year period]. / Sarcomes primitifs du sein: à propos d'une série rétrospective de 42 cas traités à l'Institut Bergonié sur une période de 32 ans.

OBJECTIVE: To evaluate the experience of a single cancer center with unusual tumors. To analyze Primary breast sarcomas (PBS). To investigate treatment and prognostic factors influencing overall survival (OS) and disease-free survival (DFS). PATIENTS AND METHODS: Retrospective study of a series of 42 patients. We reviewed the clinical records and pathology slides of 42 women with PBS treated in our institution between 1970 and 2002. Log-rank tests were used to determine OS and DFS. RESULTS: The median age at diagnosis was 56.9 years (24-81 years). Surgery was part of the therapeutic strategy in all the patients. Patients with angiosarcoma and those with malignant cystosarcoma constituted distinct populations. The 10-year OS and DFS rates were 53% and 55% for angiosarcoma patients and 89% and 100% for cystosarcoma patients (p=0.009 and 0.01 respectively). CONCLUSION: Careful preoperative multidisciplinary assessment is required before making the decision to treat. Mastectomy is generally indicated. Axillary lymph node dissection is not indicated.

Number of residual nodules, better than size, defines optimal surgery in advanced epithelial ovarian cancer.

The objective of the present study was to improve the definitions of optimal and suboptimal surgery in ovarian cancer. A retrospective prognostic factor analysis was done in a series of 433 patients with advanced ovarian cancer (stages III and IV) treated during the cisplatine era from 1980 to 1997 with assessment of postoperative residual disease by number and ranges of size of peritoneal nodules. Mean age of patients was 56.7 years. Median follow-up of patients alive was 138 months (range: 6-266 months). Median overall survival was 23 months. Significant prognostic factors for overall survival in univariate analysis were age, the presence of ascites, FIGO stage, treatment period, histological subtype, grade, results of surgery as defined by size and number of residual nodules. In multivariate analysis, quality of surgery defined by no versus few versus numerous residual nodules showed to remain an independent prognostic factor for outcome (P = 0.003), whereas size of residual nodules did not achieve significance. In conclusion, only complete surgery (no residual nodules) confers a real survival advantage. Cytoreduction to few and small nodules is associated with favorable outcome and could be qualified as optimal. Otherwise, cytoreduction leaving numerous nodules, whatever their size, remains suboptimal. Such patients should be considered for neo-adjuvant chemotherapy.

Surgical treatment of liver metastases by radiofrequency ablation, resection, or in combination.

AIMS: Radiofrequency ablation (RFA) has a role in the treatment of unresectable liver metastases either percutaneously or in open surgery. The aim of this study was to determine the feasibility and value using RFA, resection or in combination to cure liver metastases of colorectal or other origin. METHODS: Fifty-two consecutive patients were operated on with the intention to treat their liver metastases using both techniques of RFA and resection in the same curative intent. A CT scan was performed 2 months postoperatively and then every 4 months. RESULTS: Fifty patients with 137 metastases could be treated: 55 lesions were resected and 82 were ablated. Curative treatment of 13 patients could only be achieved by using RFA combined with resection. Morbidity was 16% and local treatment proved insufficient in three cases. Estimated 1-year survival probabilities were, respectively, 0.85 in the colorectal group and 0.80 in the non-colorectal group. CONCLUSIONS: RFA increased resectability of liver metastases and reduced the morbidity. Respective indications of both techniques were complementary and depend on the size and the topography of the lesion to be treated.

[Psychological adjustment of short interval follow-up mammography: about 50 cases]. / Une étude exploratoire sur le vécu psychologique des femmes lors de la surveillance des anomalies mammographiques probablement bénignes: à propos de 50 cas.

The objective was to investigate women's perceptions and the perceived stress experience when undergoing surveillance mammography for benign lesions (ACR3, BI-RADS). A semi-prospective study was carried out on women with diagnosed "probably benign" breast abnormalities. It was a multicentric study from 1st March to 10th June 2002. The survey was performed at the first follow-up mammography and included questions about perceptions and perceived stress related to the follow-up experience. The response is analysed with chi-square test. Fifty women (35-75 years) answered the questionnaire. All women were satisfied with reception. The mammographies were painful (23 cases). Subsequent time seemed to be too long (seven cases) and they were anxious in 19 cases. Quality of life was spoiled (disturbed) (20 cases). Professional and social perturbations were not frequent (four cases). Speaking to a relative is frequent (39 cases), but patients were not satisfied with it (33 cases). Medical information was estimated (47 cases) but was not satisfactory (36 cases). The median of the stress scale is 4-5. There are two pickaxes, one at 2 (11 cases), the second at 5 (8 cases). "Low-stressed" women could have an avoidance coping. "High-stressed" women could use a helplessness- hopelessness coping strategy. Women reported a good and informative medical support and adequate comprehension of the short follow-up mammography. They were reassured by the medical care, but the evaluation of the stress level shows it to be high, probably due to the uncertainty of diagnosis.

DNA topoisomerase IIalpha expression and the response toprimary chemotherapy in breast cancer.

The alpha isoform of Topoisomerase IIalpha (Topo IIalpha) is a proliferation marker as well as a target for several chemotherapeutic agents such as anthracyclines. In vitro studies have demonstrated the relationship between the Topo IIalpha expression level and chemosensitivity of target cancer cells. To verify this effect in vivo, we selected 125 patients presenting with T(2)>3 cm and T(3) N(0-1) M(0) breast tumours who were treated by six cycles of primary chemotherapy, including epirubicin before any surgery. Therapy response was assessed by clinical and X-ray mammogram measurements of tumour shrinkage. The pretherapeutic core biopsies were immunostained with a monoclonal antibody (Ki-S7) against Topo IIalpha. Ki-S7 positivity ranged from 0 to 50% (median, 15%). A high percentage of Ki-S7-positive cells (>15%) was associated with tumour regression under chemotherapy (OR=2.88, CI: 1.3-6.4, P=0.004). Ki-S7 further emerged as an independent predictor of tumour regression (OR=3.34, CI: 1.41-7.93, P=0.006), together with tumour size of less than 40 mm (OR=3.82, CI: 1.58-9.25, P=0.002) and negative oestrogen receptor (ER) status (OR=3.35, CI: 1.43-7.86, P=0.005), in a multivariate analysis including tumour size, SBR grade, ER and PR status, Ki-67, p53 and Her-2/neu. Our clinical results confirm in vitro data on the relationship between Topo IIalpha expression and tumour chemosensitivity and thus may have important practical implications.

Prognostic significance of immunohistochemically detected breast cancer node metastases in 218 patients.

Axillary lymph node metastases detected by immunohistochemistry in standard node-negative patients with breast carcinomas (13 out of 129 infiltrating ductal carcinomas and 37 out of 89 infiltrating lobular carcinomas) do not have any prognostic significance in patients followed up for a long time (respectively 24 and 18 years). Moreover, their pejorative significance in the literature is debatable since the groups and events taken into account are heterogeneous.

Application of the van nuys prognostic index in a retrospective series of 367 ductal carcinomas in situ of the breast examined by serial macroscopic sectioning: practical considerations.

UNLABELLED: The Van Nuys prognostic index (VNPI) was thought to be useful for predicting response to radiotherapy and local recurrence of ductal carcinoma in situ (DCIS). We applied the VNPI under the conditions defined by Silverstein et al., in 367 retrospective DCIS entirely sectioned into serial macroscopic 2 mm slices (155 patients had radiotherapy, median follow-up 71 months). The percentage of positive blocks with DCIS was also estimated for each specimen with cut-offs at 30% and 60% to obtain three scores. One hundred and ninety five lesions had a low VNPI, 152 an intermediate VNPI, and 20 a high VNPI. There were 9% of local recurrences (half invasive, all in the group without radiotherapy) in the low VNPI group. The local recurrence rate increased with size (p = 0.001), with reduction of distance to margins (p = 0.05), with histologic grade (p = 0.02), with percentage of positive blocks (p = 0.0003) and with VNPI score (p = 0.03). The percentage of positive blocks was the only independent predictor for local recurrence (p = 0.0001). CONCLUSION: (1) The VNPI was a local recurrence rate predictor between the low and the intermediate groups but in our series the low VNPI group had a surprisingly high local recurrence rate. (2) Only prospective studies will assess the importance of margin width and the role of radiotherapy in maintaining local control. (3) Estimation of the percentage of positive blocks is simple, may be an alternative when measurement of DCIS is difficult and should be taken into account.

Outcome of squamous cell carcinoma of the gingiva: a follow-up study of 83 cases.

BACKGROUND: Squamous cell carcinomas of the gingiva are relatively rare tumours. Standard treatment is based on surgery and radiotherapy. The extent of bone involvement affects mandibulectary indications. PURPOSE: A retrospective review of squamous cell carcinomas of the gingiva was performed to evaluate the incidence of mandibular or maxillary bone involvement. Indications for marginal and segmental bone resections are specified. MATERIAL: From 1985 to 1996, 83 patients with squamous cell carcinoma of the gingiva were treated at the Department of Surgery (Institut Bergonié, Bordeaux, France) and at the Department of Maxillofacial and Plastic Surgery (Centre Hospitalier Universitaire, Bordeaux, France). Forty-three underwent surgery plus postoperative radiotherapy. Twenty-two had flap reconstructions. Clinical evaluation and panorex roentgenography were the means used to evaluate bony invasion and to decide on the extent of bone resection. METHODS: A retrospective review of 83 consecutive patients was performed. This series is unusual in its homogeneity: surgery was performed by only two individuals and the radiotherapy was the responsibility of just two physicians. Outcome was calculated using the Kaplan-Meier method. RESULTS: Primary local control was achieved in 72 patients (87%). Overall survival and rate of recurrence were comparable to those of other squamous cell carcinomas of the oral cavity and oropharynx. CONCLUSION: Surgical resection continues to be the mainstay of treatment and this study tends to confirm the validity of modified neck dissection and marginal bone resection in suitably selected patients.

[Prosthetic breast reconstruction and radiotherapy: analysis of 67 cases)]. / Reconstruction mammaire prothétique et radiothérapie: analyse de 67 cas.

Authors inquire into repercussions of radiotherapy (RT) on prosthetic mammary reconstruction (PMR) by using the retrospective analysis of 67 cases, 59 of which were performed in irradiated areas. The particular aspect of irradiated prosthesis is evoked regarding 8 cases, and with the support of a literature review. Surgical complications and late results are evaluated comparatively to 339 PMR among non irradiated patients. According to available data in our series, harmfulness of RT not appeared statistically demonstrated. From both their experience and the literature, authors clarify indications of PMR following RT. They conclude to the necessity of a high quality RT, and optimal adaptation of surgical technique to tissular conditions.

Obvious peritumoral emboli: an elusive prognostic factor reappraised. Multivariate analysis of 1320 node-negative breast cancers.

This study was conducted to determine the prognostic influence of obvious peritumoral vascular emboli as prospectively determined by a simple routine slide examination in patients with operable node-negative breast cancer. Obvious peritumoral emboli (OPE) were defined by the presence of neoplastic emboli within unequivocal vascular lumina (including both lymphatic spaces and blood capillaries) in areas adjacent to but outside the margins of the carcinoma. OPE were assessed routinely on 5 microns thick haematoxylin and eosin-stained sections for each of 1320 primary operable node-negative breast cancers from 1975 to 1992 at our institution. OPE and other prognostic variables (tumour size, SBR grade, oestrogen and progesterone receptor status) were correlated to overall survival (OS) and metastasis-free interval (MFI) by means of univariate and multivariate analysis with a median follow-up of 103 months. OPE were found in 19.5% of tumours. In univariate analysis, OPE were related to tumour size (P = 6.3 x 10(-5)) and histologic grade (P = 4.9 x 10(-7)). Statistically significant correlations were found with OS (P = 4.6 x 10(-5)) and MFI (P = 6.4 x 10(-9)). Furthermore, in multivariate analysis, OPE was an independent prognostic variable, the most predictive factor for MFI (P = 7.7 x 10(-7)) before tumour size and grade, and was second after tumour grade for OS (P = 0.002). This study on a large unicentric series and with a long follow-up confirms the prognostic significance of vascular emboli in patients with operable node-negative breast carcinoma. Importantly, vascular emboli were found to be accurately detectable by a simple routine and non-time-consuming method. Therefore, such obvious vascular emboli should be considered as an important cost-effective, prognostic variable in patients with node-negative breast carcinoma.

Comparison of quantitative and semiquantitative methods of assessing MIB-1 with the S-phase fraction in breast carcinoma.

Different methods of assessing cell proliferation in breast cancer are currently being evaluated. Inherent qualities are required for such methods to be used on a routine basis in a pathology laboratory. Such qualities include high sensitivity and specificity in recognizing proliferating cells, simplicity in execution, and reproducibility. The MIB-1 antibody permits the immunohistochemical detection of the Ki67 antigen in fixed and paraffin-embedded tissue sections. The aim of our study was to compare a semiquantitative and quantitative method of assessing MIB-1 immunostaining with the S-phase fraction (SPF) determined by flow cytometry in a series of 112 breast carcinomas. The median semiquantitative MIB-1 score (SQ-MIB-1) in our series was 27.5%. The median quantitative MIB-1 score (B.MIB-1) was 563 positive neoplastic cells per square millimeter of tumor, and, when corrected by the volume percentage nuclei (C.MIB-1), 2844 positive nuclei per square millimeter of total nuclear area. These three indices were strongly correlated to the SPF (r = 0.73, 0.72, 0.72, n = 78), respectively for SQ.MIB-1, B.MIB-1, and C.MIB-1, MIB-1, assessed quantitatively or semiquantitatively, correlated with the Scarff, Bloom, and Richardson grade, including the mitotic index and nuclear grade, as well as with the progesterone receptor status. SQ.MIB-1 determination was easier and faster than B.MIB-1 and C.MIB-1 determination. A high correlation was found for SQ.MIB-1 results between two observers in this series (r = 0.92, n = 112), but the SQ.MIB-1 repeatability coefficient was 17.6%. Semiquantitation of MIB-1 is strongly correlated to the SPF and is an easy and rapid method of assessing cell proliferation. More studies are necessary for additional assessment of its reproducibility and its prognostic value in breast cancer.

[Breast reconstruction after breast cancer: a review of a 12-year-long experience]. / Reconstruction mammaire après cancer du sein. Bilan d'une expérience de 12 années.

The experience of 428 mammary reconstructions (MR) performed by the same surgeon, evaluated more than two years after surgery, enable the authors to try to determine possible correlations between results of MR and methods employed. Antibiotic prevention and tissue expansion only appear to have a significant influence on MR issue, whereas capsular contracture and aesthetic result did not seem correlated with prior radiotherapy or texture of the wall implant. This study confirms the low morbidity of MR. As regards quality of life, MR provides a noticeable benefit to a wide majority of patients. No method has been proved to be higher, or not advisable, against others. So, indications must be modulated in every case, patronizing better compromise between safety, quality and stability of morphologic result, and need expressed by each patient.

Direct injection/h.p.l.c. methods for the analysis of drugs in biological samples.

1. Direct injection h.p.l.c. methods for zaprinast, and pantoprazole and its sulphone metabolite were developed. 2. Optimal recovery of pantoprazole and its sulphone metabolite was effected by the absence of transfer losses and the effective adjustment of sample pH on-line. 3. Acetonitrile reduced the recovery of pantoprazole and its sulphone metabolite at acetonitrile concentrations greater than 5% in serum. 4. Direct injection h.p.l.c. methods minimize sample handling losses, reduce human contact with biological samples and are sufficiently accurate and reproducible to be used to support pharmacodynamic and toxicokinetic studies.

Determination of temelastine and a hydroxymethyl-pyridyl metabolite in biological fluid by high-performance liquid chromatography.

A high-performance liquid chromatographic method for the analysis of temelastine (1) and 2-[4-(5-bromo-3-methylpyrid-2-yl)-butylamino]-5-[6-hydroxymethy lpyrid-3- ylmethyl]-pyrimidin-4(1H)-one (1-A) in biological fluid is presented. The method combines the previously reported extraction procedure and new chromatography conditions capable of resolving 1, 1-A, and structurally similar compounds formed by the oxidation of 1. The modified method has been used to measure concentrations of 1 and 1-A in biological fluids taken from the rat and dog, and to look for the presence of 1-A in humans following administration of 1.