Deciphering the immune microenvironment of a tissue by digital imaging and cognition network.

Evidence has highlighted the importance of immune cells in various gut disorders. Both the quantification and localization of these cells are essential to the understanding of the complex mechanisms implicated in these pathologies. Even if quantification can be assessed (e.g., by flow cytometry), simultaneous cell localization and quantification of whole tissues remains technically challenging. Here, we describe the use of a computer learning-based algorithm created in the Tissue Studio interface that allows for a semi-automated, robust and rapid quantitative analysis of immunofluorescence staining on whole colon sections according to their distribution in different tissue areas. Indeed, this algorithm was validated to characterize gut immune microenvironment. Its application to the preclinical colon cancer APCMin/+ mouse model is illustrated by the simultaneous counting of total leucocytes and T cell subpopulations, in the colonic mucosa, lymphoid follicles and tumors. Moreover, we quantify T cells in lymphoid follicles for which quantification is not possible with classical methods. Thus, this algorithm is a new and robust preclinical research tool, for investigating immune contexture exemplified by T cells but it is also applicable to other immune cells such as other myeloid and lymphoid populations or other cellular phenomenon along mouse gut.

Late toxicities and clinical outcome at 5 years of the ACCORD 12/0405-PRODIGE 02 trial comparing two neoadjuvant chemoradiotherapy regimens for intermediate-risk rectal cancer.

BACKGROUND: Outcome of intermediate risk rectal cancer may be improved by the addition of oxaliplatin during 5-fluoruracil concomitant neoadjuvant chemoradiotherapy. The purpose of this study is to analyze the main clinical results of the ACCORD12 trial (NCT00227747) in rectal cancer after 5 years of follow-up. PATIENTS AND METHODS: Inclusion criteria were as follows: rectal adenocarcinoma accessible to digital examination staged T3-T4 Nx M0 (or T2 Nx distal anterior rectum). Two neoadjuvant chemoradiotherapy regimens were randomized: CAP45 (RT 45 Gy + capecitabine) and CAPOX50 (RT 50 Gy + capecitabine and oxaliplatin). Main end point was sterilization of the operative specimen. Acute and late toxicities were prospectively analyzed with dedicated questionnaires. RESULTS: Between November 2005 and July 2008, 598 patients were included in the trial. After a median follow-up of 60.2 months, there was no difference between treatment arms in multivariate analysis either for disease-free survival or overall survival (OS) [P = 0.9, hazard ratio (HR)=1.02; 95% confidence interval (CI), 0.76-1.36 and P = 0.3, HR = 0.87; 95% CI, 0.66-1.15, respectively]. There was also no difference of local control in univariate analysis (P = 0.7, HR = 0.92; 95% CI, 0.51-1.66). Late toxicities were acceptable with 1.6% G3 anal incontinence, and <1% G3 diarrhea, G3 rectal bleeding, G3 stenosis, G3-4 pain, G3 urinary incontinence, G3 urinary retention and G3 skeletal toxicity. There was a slight increase of erectile dysfunction over time with a 63% rate of erectile dysfunction at 5 years. There was no significant statistical difference for these toxicities between treatment arms. CONCLUSIONS: The CAPOX50 regimen did not improve local control, disease-free survival and overall survival in the ACCORD12 trial. Late toxicities did not differ between treatment arms.

Analysis of patients' narratives posted on social media websites on benfluorex's (Mediator® ) withdrawal in France.

WHAT IS KNOWN AND OBJECTIVE: Websites and discussion lists on health issues are among the most popular resources on the Web. Use experience reported on social media websites may provide useful information on drugs and their adverse reactions (ADRs). Clear communication on the benefit/harm balance of drugs is important to inform proper use of drugs. Some data have shown that communication (advisories or warnings) is difficult. This study aimed to explore the Internet as a source of data on patients' perception of risk associated with benfluorex and the impact of wider media coverage. METHODS: Three French websites were selected: Doctissimo, Atoute.org considered the best-known and visited website in France for health questions and Vivelesrondes (Long live the Tubbies) for overweight people. Three periods were chosen: (1) before November 2009 (i.e. before benfluorex withdrawal), (2) between November 2009 and November 2010 (when the risk of valvulopathy with benfluorex appeared in social media) and (3) after November 2010. RESULTS AND DISCUSSION: Two hundred twenty initial postings were analysed. These lead to 660 secondary postings which were analysed separately. In period 1, 114 initial postings were analysed, mostly concerning efficacy of the drug (72%). In period 2, 42 initial postings were analysed involving mainly ADRs or warnings (73%). In period 3, 64 initial postings were analysed; most frequent expressing anger directed at the healthcare system (58%) and anxiety about cardiovascular ADRs (30%). Online consumer postings showed that there were drastic changes in consumers' perceptions following media coverage. WHAT IS NEW AND CONCLUSION: This study suggests that analysis of website data can inform on drug ADRs. Social media are important for communicating information on drug ADRs and for assessing consumer behaviour and their risk perception.

Return to work of 87 severely impaired low back pain patients two years after a program of intensive functional rehabilitation.

UNLABELLED: Return to work is the main long-term objective of rehabilitation programs for patients with chronic low back pain (LBP). OBJECTIVES: Evaluation of work status and number of sick leaves in 87 severely impaired LBP patients 2 years after a functional restoration program. PATIENTS AND METHODS: Open prospective study. POPULATION: 87 chronic LBP patients. INTERVENTION: multidisciplinary functional restoration program. Ergonomic advice on the workplace was performed for 53 patients. OUTCOME: work status and number of sick leaves due to LBP. RESULTS: The characteristics of the 26 patients lost to follow-up did not differ significantly from the rest of the population before the program. In the 61 remaining patients, 48 (78%) were at work at 2 years, 43 full-time and 22 at the same job. Nineteen worked in a different environment. Sick leaves were reduced by 60% compared to the 2 years prior to the program: 128 days (+/-200 days) versus 329 days (+/-179 days); p<0.005. CONCLUSION: Sick leaves remained significantly reduced and the number of workers who were at work significantly increased at 2 years after an intensive program.

Management of low back pain in primary care prior to multidisciplinary functional restoration: a retrospective study of 72 patients.

OBJECTIVE: Chronic low back pain is a major socioeconomic health issue, due to the high direct (healthcare) and indirect (sick leave) costs. The aim of the present study was to describe the primary care management of low back pain patients prior to their inclusion in a multidisciplinary functional restoration network. METHODS: A descriptive, retrospective, questionnaire-based survey of the general practitioners dealing with 72 low back pain patients. RESULTS: Patients had been monitored by their general practitioner for an average of four years, with a mean frequency of eight appointments per year per patient. Ninety-three percent and 60% of the patients had been referred to a rheumatologist and a surgeon, respectively. Ninety-eight percent had had lumbar radiographies, 80% had undergone a computed tomography scan and 64% had undergone magnetic resonance imaging. The most commonly prescribed medications were anti-inflammatories and first- or second-line analgesics. Thirty percent had already received morphine analgesics and 50% had taken antidepressants. Thirty-two percent had undergone lumbar surgery. Physiotherapy was frequently reported and, indeed, 6% of patients had participated in over 100 sessions. Total sick leave averaged 8.25 months over the study's follow-up period. CONCLUSION: The time interval before referral to a multidisciplinary care team is long and so GPs should be encouraged and helped to organize this process earlier. It is also essential to determine factors which predict progression to chronic LBP.

Guidelines for postmortem protocol for ocular investigation of sudden unexplained infant death and suspected physical child abuse.

Postmortem examination is a cornerstone in identifying the cause of unexplained sudden death in children. Even in cases of suspected or known abuse, an autopsy may help characterize the nature of the abuse, which is particularly important in the forensic autopsy of children in the first 3 to 4 years of life when inflicted neurotrauma is most common. Forensic examinations are vital in cases that might otherwise be diagnosed as sudden infant death syndrome. The ocular autopsy in particular may demonstrate findings that were not appreciated on antemortem clinical examination. This protocol for postmortem examination of the eyes and orbits was developed to promote more consistent documentation of findings, improved clinical and forensic decision making, and more replicable and coherent research outcomes.

[Evaluation of the handicap and the quality of life in spinal cord injuries: study in a population of 58 patients living at home]. / Evaluation du handicap et de la qualité de vie chez les blessés médullaires: étude d'un échantillon de population de 58 sujets vivant à domicile.

GOAL: The main purpose of this study was to identify objective factors (social and economic status, impairments, functional limitations) contributing to the subjective quality of life of subjects presenting with residual neurological deficits from a traumatic spinal cord injury (SCI) and living at home. SUBJECTS AND METHOD: This is a cross-sectional descriptive study of a sample of SCI patients followed at a Physical Medicine and Rehabilitation facility. After informed consent was obtained, a clinical and functional examination was conducted and questionnaires were filled out by the subjects. The following parameters were assessed: impairments (sensory and motor ASIA scores), disability (Barthel Index and Functional Independence Measure), and quality of life (Reintegration to Normal Living Index and Nottingham Health Profile). RESULTS: Fifty-eight subjects completed the study (mean age 41.38 +/-13.55 years and mean delay from onset 6.24 +/- 6.06 years). There was a strong significant correlation between the level of quality of life and the current age, the age at the time of the accident and the disability level, particularly for the dimensions of physical independence, social integration and mobility. CONCLUSION: Our results are consistent with published data, which identify age as a determining factor of quality of life after SCI. Our review of the literature indicated that time from onset influences quality of life. Longitudinal studies, taking into account the levels of social participation, are needed to better understand the impact of time on quality of life after SCI.

Amino acids: analytical aspects.

This workshop was organized as a direct response to concerns and queries raised by laboratory personnel, both in Europe and in the United States, about the imminent withdrawal of Beckman Coulter from the amino acid analysis market. The topics covered included external quality control schemes, standard operating procedures for amino acid analysis and instrumentation, both from a user's perspective and that of the company representatives. There was a discussion panel of all speakers following the presentations.

In vitro activity of quinupristin/dalfopristin in comparison with five antibiotics against worldwide clinical isolates of staphylococci.

OBJECTIVE: To evaluate the in vitro activity of quinupristin/dalfopristin (Q/D), a streptogramin combination, in comparison with five antibiotics against worldwide clinical isolates of staphylococci. METHODS: A multicenter in vitro study was performed using the E test during a period of 3 months (April to June) in 1997 on fresh, clinically significant, non repetitive strains of staphylococci from patients hospitalized in 23 different hospitals in 18 countries tested. RESULTS: A total of 2132 staphylococcal isolates including methicillin resistant (MR), methicillin susceptible (MS) S. aureus (1003 MS, 462 MR), S. epidermidis (169 MS, 251 MR), S. haemolyticus (28 MS, 46 MR), S. hominis (28 MS, 16 MR), and coagulase negative staphylococci (86 MS, 43 MR) were analyzed. Q/D was highly active against all species tested. MIC90 (mg/L) ranged from 0.5 to 2 depending on the species. Strains had MIC < or = 1 mg/L in 97.6%. For S. aureus, S. epidermidis, S. hominis and other coagulase-negative staphylococci no differences in MIC90 were observed for MS or MR. One dilution difference was observed for S. haemolyticus, which overall was the less susceptible species. Erythromycin resistance was observed among 57- 87% of MR-strains and was lower among MS-strains (18-56%). Erythromycin resistance had no or little influence on MIC of Q/D. In comparison to vancomycin, Q/D was two to four times more active. CONCLUSIONS: The streptogramin combination Q/D showed an excellent in vitro activity against all staphylococcal species tested regardless of the resistance pattern to other drug classes, particularly resistance to methicillin. Q/D was two to four times more active than vancomycin and MIC values varied from 0.5-2 according to the species. The synergy of Q/D was well conserved in macrolide-resistant strains.

Lidocaine plus ropivacaine versus lidocaine plus bupivacaine for peribulbar anesthesia by single medical injection.

UNLABELLED: This study was designed to compare the effects of ropivacaine and bupivacaine, each combined with lidocaine, during peribulbar anesthesia by single medial injection for cataract surgery. One hundred patients were included and randomly divided into two groups of 50, given a mixture of 50% bupivacaine (0.5%) and 50% lidocaine (2%) or 50% ropivacaine (1%) and 50% lidocaine (2%), and 25 U hyaluronidase per mL with each combination. After the first injection, patients given ropivacaine exhibited significantly better akinesia than those given bupivacaine, and significantly fewer were reinjected (19/50 vs 31/50). Among the patients reinjected, peroperative akinesia and analgesia proved satisfactory in both groups. We observed three cases of diplopia caused by retraction of the internal rectus muscle and two cases of moderate ptosis after superonasal reinjection. Hemodynamic profiles were similar in the two groups, and no major side effects were noted during the observation. One percent ropivacaine may be a more appropriate agent than 0.5% bupivacaine for peribulbar anesthesia by single medial injection. IMPLICATIONS: One percent ropivacaine may be a more appropriate agent than 0.5% bupivacaine for peribulbar anesthesia by single medial injection. Combined with lidocaine, it provides better akinesia and similar analgesia.

Suitability of MDCK cells grown in a serum-free medium for influenza virus production.

MDCK cells have been adapted to grow in a serum-free environment using Ultra-MDCK medium (BioWHITTAKER). The growth of adapted U-MDCK cells was maintained for over a year without any reduction in growth rate or modification of cell karyotype; cells were scaled up to spinner culture using several microcarriers. The cells were shown to be a very good host for influenza A and B virus replication in both the presence and absence of trypsin in the infection medium. Trypsin-independent viruses replicated to high titres (10(7)-10(8) TCID50/ml) in U-MDCK cells, after selection through serial passages without trypsin. This virus progeny exhibited uncleaved and antigenically modified haemagglutinin compared with standard viruses grown with trypsin. Finally, large amounts of influenza A and B viruses were produced in U-MDCK cells grown on microcarriers under rod-stirred conditions using selected trypsin-independent variants.

Inclusive dielectron cross sections in p + p and p + d interactions at beam energies from 1.04 to 4.88 GeV.

Measurements of dielectron production in p + p and p + d collisions with beamkinetic energies from 1.04 to 4.88 GeV are presented. The differential cross section is presented as a function of invariant pair mass, transverse momentum, and rapidity. The shapes of the mass spectra and their evolution with beam energy provide information about the relative importance of the various dielectron production mechanisms in this energy regime. The p + d to p + p ratio of the dielectron yield is also presented as a function of invariant pair mass, transverse momentum, and rapidity. The shapes of the transverse momentum and rapidity spectra from the p + d and p + p systems are found to be similar to one another for each of the beam energies studied. The beam energy dependence of the integrated cross sections is also presented.

What have been the strategies for the registration, positioning and control of medical information for intramuscular artemether?

At the beginning of the 1990s the Chinese Authorities and Rhone-Poulenc Rorer signed an agreement to develop intramuscular (i.m.) artemether (Paluther) and to market the product in the malaria endemic countries. This accord ushered in an exemplary period of co-operation between an international pharmaceutical group and its Chinese partners, the WHO (especially TDR), the Wellcome Trust, and several university research departments. The challenge was to complement the Asian development (to Western standards of Good Manufacturing Practices, Good Laboratory Practices, and Good Clinical Practices) of a molecule which was already used on an everyday basis in East Asia and by Chinese medical missions in Africa. The implementation of Good Manufacturing Practice was the priority for Rhone-Poulenc Rorer in order to ensure the pharmaceutical quality of Paluther. New preclinical and clinical studies confirmed the importance of the drug in the curative treatment of severe malaria due to Plasmodium falciparum, or when resistance to other antimalarial drugs is suspected. The outcome of these new trials was the recognition that i.m. artemether is at least as efficient as quinine. The results of the current development of Paluther have been presented at several international congresses and the latest clinical trials were published in the New England Journal of Medicine in July 1996. The neurotoxicity observed in animals after long term administration of high and repeated dosages has never been reported in human subjects. I.m. artemether was listed in the WHO List of Essential Drugs in December 1995, and the product has now been registered in more than 40 malaria endemic countries. Authorization for use of Paluther in hospitals in France and in several other European countries was granted in 1996.

Preoperative color-Doppler assessment of vascularisation of the rectus abdominis: anatomic basis of breast reconstruction with a transverse rectus abdominis myocutaneous flap--a prospective study.

The unipedicled TRAM flap is an useful alternative to breast reconstruction after mastectomy in patients who refuse mammary implants. There is however the risk of unpredictable partial skin necrosis even after rigorous surgical procedures. Certain authors have proposed color flow doppler assessment before reconstructive surgery better to identify the vascular network and optimise patient selection. We performed a prospective study in 20 outpatients in order to compare preoperative assessment of the blood supply to the abdominal flap with the operative findings. An Ultramark 9 HDI (Advanced Technology Laboratories) equipped with a high frequency (10 Mhz) linear probe was used to measure blood flow and vessel caliber in the epigastric a. and perforating vessels (localisation, number; peak flow). Despite three limiting factors (anatomic, technical, morphologic) the results obtained in this series could be used to determine the indications for a TRAM flap.

The view of the pharmaceutical industry.

Rhône-Poulenc Rorer has committed itself to the development of artemether because we believe the drug will be of considerable benefit to sufferers from severe falciparum malaria, and because it is a stable, effective and economical compound that can be given by intramuscular injection. The quality of the pharmaceutical product meets international regulatory standards. Artemether is unlikely to yield big profits, but we believe that major pharmaceutical companies have a responsibility to develop such much-needed products. To develop this project further, we will need the assistance of academic institutions, research organizations and international bodies.