Mortality and morbidity peaks modeling: An extreme value theory approach.

Hospitalizations and deaths belong to the most studied health variables in public health. Those variables are usually analyzed through mean events and trends, based on the whole dataset. However, this approach is not appropriate to comprehend health outcome peaks which are unusual events that strongly impact the health care network (e.g. overflow in hospital emergency rooms). Peaks can also be of interest in etiological research, for instance when analyzing relationships with extreme exposures (meteorological conditions, air pollution, social stress, etc.). Therefore, this paper aims at modeling health variables exclusively through the peaks, which is rarely done except over short periods. Establishing a rigorous and general methodology to identify peaks is another goal of this study. To this end, the extreme value theory appears adequate with statistical tools for selecting and modeling peaks. Selection and analysis for deaths and hospitalizations peaks using extreme value theory have not been applied in public health yet. Therefore, this study also has an exploratory goal. A declustering procedure is applied to the raw data in order to meet extreme value theory requirements. The application is done on hospitalization and death peaks for cardiovascular diseases, in the Montreal and Quebec metropolitan communities (Canada) for the period 1981-2011. The peak return levels are obtained from the modeling and can be useful in hospital management or planning future capacity needs for health care facilities, for example. This paper focuses on one class of diseases in two cities, but the methodology can be applied to any other health peaks series anywhere, as it is data driven.

[Fear of childbirth among nulliparous women: Relations with pain during delivery, post-traumatic stress symptoms, and postpartum depressive symptoms]. / La peur de l'accouchement chez des nullipares: lien avec la douleur lors de l'accouchement, les symptômes de stress post-traumatique et les symptômes dépressifs post-partum.

OBJECTIVE: Fear of childbirth is common in women who are pregnant with their first child and is associated with important consequences such as abortions and miscarriages. Twenty percent of nulliparous women seem to exhibit a mild or moderate fear, while 6% present an excessive and irrational fear known as tocophobia. Tocophobia is suggested to be associated with many negative consequences such as postpartum depression (PPD) and Post-traumatic stress (PTS). However, there is little empirical evidence to support these relationships. Recently, Fairbrother and Woody (2007) did not observe a link between the fear of childbirth and symptoms of PPD and PTS in nulliparous women. Some results, near the significance level, could be explained by a lack of statistical power. The present study focused on the link between the fear of childbirth and the process of delivery, the perception of pain, PPD and PTS. More specifically, it aimed to test three hypotheses: (i) fear of childbirth will be linked to the process of delivery, especially regarding the perception of pain, the use of anaesthesia and the use of Caesarean section; (ii) a high level of fear of childbirth will be associated with more negative postpartum consequences (namely PPD/PTS symptoms); (iii) the process of delivery and pain will also be related to post-delivery symptoms. Mediation effects were tested. METHOD: Data from a longitudinal study were used to meet the hypotheses. A total of 176 nulliparous pregnant women responded to questionnaires at two time measurements (during pregnancy and at 5weeks postpartum). RESULTS: Fear of childbirth is related to the perception of pain at birth among women delivering vaginally, in the absence of anaesthesia. It is also linked to symptoms of PPD and PTS, regardless of whether or not anaesthesia was used. Fear of childbirth also appears to be strongly associated to symptoms of PTS in women who have experienced an unplanned caesarean section. Thus, symptoms of postpartum PTS could play a mediating role in the link between fear of childbirth and PPD. CONCLUSIONS: These results support the relevance of taking into account the fear of childbirth and perception of pain in connection with symptoms of PTS and PPD in nulliparous women. The unplanned caesarean section (including emergency caesarean) also appears to be important in the study of the relationship between fear and symptoms of PTS. Fear of childbirth could render the experience of childbearing more negative and predispose to PTS and PPD. Enabling psychological vulnerabilities could also be an interesting avenue for understanding these links. Limitations are discussed.

The systemic control of circadian gene expression.

The mammalian circadian timing system consists of a central pacemaker in the brain's suprachiasmatic nucleus (SCN) and subsidiary oscillators in nearly all body cells. The SCN clock, which is adjusted to geophysical time by the photoperiod, synchronizes peripheral clocks through a wide variety of systemic cues. The latter include signals depending on feeding cycles, glucocorticoid hormones, rhythmic blood-borne signals eliciting daily changes in actin dynamics and serum response factor (SRF) activity, and sensors of body temperature rhythms, such as heat shock transcription factors and the cold-inducible RNA-binding protein CIRP. To study these systemic signalling pathways, we designed and engineered a novel, highly photosensitive apparatus, dubbed RT-Biolumicorder. This device enables us to record circadian luciferase reporter gene expression in the liver and other organs of freely moving mice over months in real time. Owing to the multitude of systemic signalling pathway involved in the phase resetting of peripheral clocks the disruption of any particular one has only minor effects on the steady state phase of circadian gene expression in organs such as the liver. Nonetheless, the implication of specific pathways in the synchronization of clock gene expression can readily be assessed by monitoring the phase-shifting kinetics using the RT-Biolumicorder.

New immediate release formulation for deterring abuse of methadone.

CONTEXT: Drug abusers are known to take a dosage form containing an opioid analgesic and crush, shear, grind, chew, or dissolve it in water or in alcohol, in order to extract the opioid component. OBJECTIVE: Develop an anti abuse immediate release formulation using methadone as model drug. MATERIALS AND METHODS: Tablets combining methadone and alkalizing agents were manufactured. A methadone assay was used to determine extraction efficiency from tablets in aqueous and alcohol solvents. In vitro dissolution testing was used to determine drug release in different media. RESULTS AND DISCUSSIONS: Meglumine-based formulations prevented extraction of 70 to 100% of methadone from tablets. Addition of this alkalizing agent caused methadone to precipitate out of a solution along with other ingredients and be retained on standard filters. Meglumine-containing and control tablets showed similar dissolution profiles in acidic media, suggesting adequate solubilisation of the drug early in the gastrointestinal tract. Finally, stability upon storage of the formulations for 6 months at 25°C/60%RH and 40°C/75%RH was confirmed. CONCLUSION: Incorporation of an alkalizing agent into methadone tablets significantly reduced the preparation of a methadone solution for intravenous administration and abuse, while allowing the formulation to release methadone in gastric media and provide desired pharmacological effect.

[French adaptation and validation of a scale measuring the fear of childbirth]. / Évaluation de la peur de l'accouchement. Validation et adaptation française d'une échelle mesurant la peur de l'accouchement.

OBJECTIVES: The fear of childbirth, a central aspect of tokophobia, recently started to capture the attention of the scientific community as a potential determinant of obstetric and post-natal complications. However, studies on this subject are still few and this can be partly explained by the lack of validated instruments, especially in French. This paper presents the results from two studies designed to develop and evaluate the psychometric properties of a French version of the Traumatic Event Scale (TES), adapted to assess fear of childbirth (Söderquist et al., 2004 [21]). METHOD: The first study presents details regarding the development of this scale and checks the quality of the resulting items as well as their internal consistency, convergent validity and factorial validity. This study relied on a sample of 65 mothers with at least one child under the age of 36 months. In the second study, the psychometric properties of the instrument developed in Study 1 were tested more systematically on a sample of 204 women who were at the time experiencing their first pregnancy. RESULTS AND CONCLUSION: The results from the first study show adequate psychometric properties, strong correlations with measurements assessing worry, and support a five factor model. Results from this second study replicated the results from the first one on the basis of confirmatory factor analyses. Findings presented in these studies confirm that this instrument presents very good psychometric properties as a measurement of the fear of childbirth in pregnant women.

Secular trends of hip fractures in Québec, Canada.

UNLABELLED: This study examined the secular trends of hip fracture incidence among individuals 50 years and older in Québec between 1993 and 2004. Age-standardized rates decreased at both the provincial and regional levels. The largest relative decrease was observed among younger females, and rates declined more slowly in the elderly. INTRODUCTION: The population of the province of Québec is among the oldest in North America. Before the trend rupture reported in the late 1990s in several countries, hip fracture (HF) incidence rates did not show a secular trend (between 1981 and 1992). This study examined the secular trends of HF incidence at the provincial level and in two of the most important urban areas of the province, Montréal and Québec City, between 1993 and 2004. METHODS: All hospitalisations of individuals 50 years and older living in the province of Québec between 1993 and 2004 with a main diagnosis of HF were included. Standardized rates of HF incidence were calculated for females and males, 50-74 years and 75 years and older. RESULTS: The Québec City area showed a strong decreasing trend in HF rates for younger females, but the other groups did not show an obvious trend. Although our models did not support the existence of significant differences in trends between both areas, the rates of HF of younger males and, to a lesser extent, of older women in the Montréal area were significantly higher than in the Québec City area. CONCLUSIONS: Differences observed in hip fracture rates as well as in secular trends between age groups and gender emphasise the need for decision makers to rely on results based on age-specific and sex-specific analyses.

Environmental characteristics associated with campylobacteriosis: accounting for the effect of age and season.

Campylobacteriosis is a leading cause of acute bacterial gastroenteritis. An ecological study was undertaken to explore the association between environmental characteristics and incidence of campylobacteriosis in relation to four age groups and two seasonal periods. A multi-level Poisson regression model was used for modelling at the municipal level. High ruminant density was positively associated with incidence of campylobacteriosis, with a reduced effect as people become older. High poultry density and presence of a large poultry slaughterhouse were also associated with higher incidence, but only for people aged 16-34 years. The effect of ruminant density, poultry density, and slaughterhouses were constant across seasonal periods. Other associations were detected with population density and average daily precipitation. Close contacts with farm animals are probably involved in the associations observed. The specificity of age and season on this important disease must be considered in further studies and in the design of preventive measures.

[Etiological factors of generalized anxiety disorder]. / Les facteurs étiologiques du trouble d'anxiété généralisée: état actuel des connaissances sur les facteurs psycho-sociaux.

Generalized Anxiety disorder (GAD) is a widespread psychiatric syndrome involving significant consequences on people's health. However, recent data show that this disorder has received little attention when compared to other anxiety disorders. A review of the publication on GAD also stated that the majority of research has been conducted on descriptive issues, whereas etiological factors received lower rates of publication. This may be explained by the fact that changes in the conceptualization of GAD slowed down the progression of researches focussing on the identification of factors implicated in the development and the maintenance of its symptoms. For example, in the third version of the Diagnostic and Statistical Manual of Mental disorders (DSM) published in 1980, GAD could only be assigned if patients had a persistent anxiety without reporting specific symptoms of other anxiety disorders (eg Specific Phobias, Panic Disorder, Obsessive-Compulsive Disorder). With DSM III-R and DSM IV, excessive worry became the central feature of GAD, which increased its diagnostic validity. Taking that into account, where do we stand in the comprehension of GAD? What are the etiological factors identified as playing a role in the development and the maintenance of GAD and its central feature, excessive and uncontrollable worry? The goal of this article consists to provide answers to these questions by reviewing the literature of the current knowledge of GAD and examining various theories and explanatory models available. Firstly, many studies suggest that environmental factors are implicated in the development and maintenance of GAD. However, the majority of researches rests on an out-of-date conceptualization of GAD. Furthermore, the nature of experimental designs used in researches limits their conclusion. The first environmental factor documented is negative life events. Researchers noted that the occurrence of one or more negative life events being significant and appearing in an unexpected way was associa-ted with an increase in the risk to develop a GAD (eg. Familial problems (eg conflicts, abuses), considered as chronic stress-inducing situations, have also been suggested as etiological factor of GAD. Among the other environmental factors identified are separation during childhood, role inversion during childhood, lack of social interactions, poor life satisfaction, and modeling of a relative having an anxiety disorder. The cognitive and behavioral approach has been widely used during the last two decades to precise the etiology of GAD. First of all, some studies have pointed out the function of worry, which is to suppress images associated with negative thoughts (eg. Thus, GAD patients seem to develop excessive worries in order to prevent more negative feeling associated with the images of a thought. Other authors suggested that a deficit of attention might be responsible of excessive worry. Finally, some cognitive variables such as intolerance of uncertainty, false beliefs about worry, negative problem orientation and cognitive avoidance have been suggested as playing a determinant role in the development and maintenance of GAD. In order to resume the empirical findings available in the literature, an etiological model gathering the cognitive, behavioral, and environmental factors implicated in GAD is proposed. This model suggests that GAD is the result of a psychological vulnerability activated by a tension resulting from a negative event. Finally, recommendations are brought concerning future researches on GAD and its central characteristic, excessive and uncontrollable worry.

Echinoderms as bioindicators, bioassays, and impact assessment tools of sediment-associated metals and PCBs in the North Sea.

The study assessed the occurrence, possible toxicity, and impact of sediment-associated metals and PCBs in the coastal zone of the southern North Sea using echinoderms as representatives of the macrobenthos. Metals and PCBs were analyzed in the sediments and in the body compartments of the starfish Asterias rubens from 11 stations. The general toxicity of sediment-associated contaminants was assessed by bioassays using embryonic and larval developments of both A. rubens and the sea urchin Psammechinus miliaris. The impact of contamination was assessed by measuring cellular immune responses of A. rubens collected in the same stations. Contamination of the starfish by metals and PCBs closely reflected that of the sediments. However, bioaccumulation was element-specific for metals and depended on the chlorination pattern for PCBs. The sediment-associated contaminants appeared to be toxic in both the A. rubens and P. miliaris developmental assays. Moreover, metals were shown to affect the immune responses of starfishes living in contaminated stations. The most significant effects on biological responses were recorded in the plumes of the Scheldt/Rhine/North Sea Canal and the Elbe/Weser Rivers.

Polymorphic properties of micronized carbamazepine produced by RESS.

Carbamazepine microparticles were produced by the rapid expansion of supercritical carbon dioxide solutions (RESS) method. The characteristics of the resulting particles were studied by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and image analysis. X-ray diffractograms and SEM photomicrographs revealed that the crystalline nature of the produced carbamazepine microparticles depended on operating pressure and temperature conditions. Different polymorphs were obtained under various operating conditions. Under certain temperature (below 40 degrees C) and pressure (below 240 bar) conditions, it was possible to form primarily the carbamazepine polymorph stipulated by US Pharmacopeia. A significant reduction was observed in the particle size and size distribution range of carbamazepine produced by RESS. The processed particles had a mean diameter smaller than 3 microm and a size distribution range between 0.5 and 2.5 microm compared to unprocessed starting material with a mean diameter of approximately 85 microm and a size distribution range between 15 and 336 microm. Thus, this study demonstrates that the polymorphic characteristics of carbamazepine microparticles produced by the RESS method can be controlled by varying operating pressure and temperature parameters.

[Study of psychometric properties of two new questionnaires assessing beliefs in hypochondriasis in a non-clinical population]. / Evaluation des croyances reliées à la santé. Etude des propriétés psychométriques de deux nouveaux instruments evaluant les croyances présentes dans l'hypocondrie auprés d'une population non clinique.

Most instruments focussing on hypochondriasis symptoms do not have for goal to assess beliefs specifically. Instead, these instruments are used to measure specific behaviors. To assess underlying beliefs with these kinds of instruments, you have to extract false beliefs by deduction. In cognitive therapy, it is important to target erroneous beliefs in order to change them. On the other hand, existing instruments are not really suitable to target erroneous health beliefs. Even if some questionnaires are built to assess beliefs directly, it seems that they only measure the conviction of having an illness and do not assess the general health beliefs present in excessive health worriers. However, many researchers argue that this other kind of beliefs are the ones responsible in maintaining hypochondriasis symptoms. Presently, researchers assume that erroneous beliefs can maintain worries about illness among people with hypochondriasis symptoms like false beliefs about worry maintain worries in people with General Anxiety Disorder (GAD). Even if the importance of false beliefs in the maintenance of pathological worries is now recognized, most instruments on hypochondriasis symptoms do not have for goal to assess erroneous beliefs concerning worry about health. For instance, although the questionnaire Why do people worry? (WW) shows good psychometric properties and measures beliefs related to general worries, this questionnaire is not specific enough to correctly evaluate beliefs associated to health worry. A new questionnaire has to emerge in order to assess false beliefs associated to worry about health. This manuscript presents the development and the validation of a new questionnaire: the General Health Beliefs Questionnaire (GHBQ) that assesses general health beliefs, and also presents the development and the validation of a new questionnaire assessing beliefs associated to worry about health: the Why do people Worry about Health? (WW-H) . In this study, the GHBQ's and the WW-H's psychometric qualities and the factorial structure were assessed. More precisely, this study examined the factorial structure, the temporal stability, the convergent, divergent and criteria validities of the GHBQ and the WW-H. Four hundred and twenty nine French-speaking university students (non-clinical participants) completed a battery of questionnaires at the beginning of a class. The questionnaires were: The General Health Beliefs Questionnaire (GHBQ), the Why do people Worry about Health (WW-H), the Illness Worry Scale (IWS), the Beck Depression Inventory-short form (BDI-short form) and the Beck Anxiety Inventory (BAI). A second administration took place three weeks later with the same sample to test the temporal stability of the GHBQ and the WW-H. The principal component analysis with orthogonal rotation (varimax) supports a five components solution for the GHBQ: 1) magical thinking, 2) health, 3) consequences, 4) responsibility, and 5) vulnerability. The principal component analysis with oblique rotation (direct oblimin) (d=0) found a two components solution for the WW-H: 1) utility and 2) magical thinking associated to worries about health. The internal consistency of the GHBQ and the WW-H is excellent (a=.80 and a=.90, respectively). A correlation of 0.49 was found between the GHBQ and the WW-H. The correlation between the GHBQ and the IWS who evaluates the tendency to worry (r=.50) and between the WW-H and the IWS (r=.49) showed that the convergent validity of these questionnaires is adequate. On a three weeks interval, the GHBQ (r=.70) and the WW-H (r=.71) showed a satisfying temporal stability. The means of the high worriers (80 superior percentile at the IWS) (M=38.8, ET=8.93) and the means of the moderate worriers (between the 40 and the 60 percentile at the IWS) (M=32.8, ET=8.00) on the GHBQ have been compared. A significant difference has been found between the two groups [F(1,181)=23,129, p<0,001]. Also, the means of the high worriers (M=27.3, ET=8.59) and the means of the moderate worriers (M=23.8, ET=8.56) on the WW-H haveave been compared. An ANOVA has found a significant difference between these two groups [F(1,180)=7,396, p=0,007]. These results show that general health beliefs and false beliefs associated to worry about health are more often present in high worriers than in moderate worriers. The GHBQ and the WW-H allow psychologists, physicians and psychiatrists to do a quicker and more exhaustive evaluation of general health beliefs and false beliefs associated to worry about health, in less costs. These questionnaires will improve the chances of success of the hypochondriasis' treatment by helping clinicians to detect and correct false beliefs more easily.

Renormalization group in quantum mechanics at zero and finite temperature.

We apply the renormalization group formalism, to integrate quantum fluctuations of quantum mechanical systems at zero and finite temperature. At zero temperature a nonperturbative renormalization group equation allows to compute the ground state energy whereas at finite temperature a variational renormalization group equation is proposed to compute the free energy.

[Evaluation of worry: validation of a French translation of the Penn State Worry Questionnaire]. / Evaluation des inquiétudes: validation d'une traduction française du Penn State Worry Questionnaire.

UNLABELLED: Excessive worry, which is the central feature of Generalized Anxiety Disorder (GAD), is recognized as an important clinical phenomenon with many negative consequences on people's health. For example, studies report that excessive worry is associated with higher frequencies of work absenteeism and medical consultations, increased risk of other anxiety disorders, depression, heart disease, diabetes and cancer. It is thus imperative to develop effective measures to assess worry among adult. Until now, no measures were available in French for the practitioners and researchers to evaluate the tendency to engage in excessive and uncontrollable worry. One of the most frequently used questionnaires to assess worry in English is The Penn State Worry Questionnaire (PSWQ). This self-report measure has been widely studied and has shown excellent psychometric properties among the non-clinical and clinical populations. This manuscript presents the results of three studies evaluating the psychometric properties of a French Translation of the PSWQ, the Questionnaire sur les Inquiétudes de Penn State (QIPS). The first study includes the translation procedures used to create the French version of the questionnaire, the factorial structure, the internal consistency, the quality of the items, and the convergent validity among non-clinical participants. The second study examines the temporal stability, and the convergent and divergent validity of the questionnaire. Finally, the third study investigates the internal consistency, the quality of the items, and the convergent and divergent validity of the questionnaire among GAD patients. Study 1. This study describes the translation procedures used to create the QIPS, the factorial structure, the internal consistency, the quality of the items, and the convergent validity among non-clinical participants. The French version was translated by a group of clinical psychologists with the assistance of a professional translator and a linguist. A back translation procedure was also conducted. Finally, a pilot study confirmed the intelligibility of the questionnaire. French-speaking university students (N = 352) completed a battery of questionnaires during a lecture. The questionnaires were: The Questionnaires sur les Inquiétudes de Penn State, the Intolerance of Uncertainty Questionnaire, the Cognitive Avoidance Questionnaire, the Why Worry Questionnaire-Revised, and the Social Problem-Solving Inventory-Revised. The statistical analyses reveal that the QIPS shows an appropriate factorial structure, an excellent internal consistency and a very good convergent validity. This French Questionnaire thus seems to be suitable to assess the tendency to worry among the nonclinical population. Study 2. This second study examines the temporal stability, and the convergent and divergent validity of the QIPS. The sample includes 95 French-speaking University students who volunteered to complete the QIPS and the Beck anxiety Inventory. Two self-rating questions were also asked to evaluate the percentage of time spent worrying and to what extent does worrying causes a problem. Eighty-six students participated at the second administration of the questionnaires, four weeks later. The results indicate that the QIPS is very stable over time, thus supporting its temporal stability. Different correlations confirm its convergent and divergent validity. It is concluded that the QIPS is a suitable measure of worry in a non-clinical population. Study 3. This study investigates the internal consistency, the quality of the items, and the convergent and divergent validity of the questionnaire among GAD patients. Seventy-seven GAD patients, as defined by the Diagnostic and Statistical Manual of Mental disorders, fourth edition (DSM IV), participated in this study. They completed the QIPS, the Worry Domains Questionnaire, The Worry and Anxiety Questionnaire, the Intolerance of Uncertainty Questionnaire, the Beck Anxiety Inventory, the Beck Depression Inventory, and a daily diary of differents aspects of worry. The results reveal that the QIPS has an excellent internal consistency, a very good convergent and divergent validity. The QIPS thus seems to be suitable to assess the tendency to worry among GAD patients. CONCLUSION: The overall results presented in these studies confirm the excellent psychometric properties of the QIPS. This French questionnaire is a useful tool for clinicians and researchers to assess the tendency to worry among non-clinical and clinical populations.

Facial morphology and children's categorization of facial expressions of emotions: a comparison between Asian and Caucasian faces.

The effects of Asian and Caucasian facial morphology were examined by having Canadian children categorize pictures of facial expressions of basic emotions. The pictures were selected from the Japanese and Caucasian Facial Expressions of Emotion set developed by D. Matsumoto and P. Ekman (1989). Sixty children between the ages of 5 and 10 years were presented with short stories and an array of facial expressions, and were asked to point to the expression that best depicted the specific emotion experienced by the characters. The results indicated that expressions of fear and surprise were better categorized from Asian faces, whereas expressions of disgust were better categorized from Caucasian faces. These differences originated in some specific confusions between expressions.

Experimental manipulation of intolerance of uncertainty: a study of a theoretical model of worry.

Intolerance of uncertainty has been identified as an important variable related to worry and Generalized Anxiety Disorder (GAD) [Dugas, M. J., Gagnon, F., Ladouceur, R., & Freeston, M. H. (1998). Generalized anxiety disorder: a preliminary test of a conceptual model. Behaviour Research and Therapy, 36, 215-226; Ladouceur, R., Dugas, M. J., Freeston, M. H., Rhéaume, J., Blais, F., Boisvert, J.-M., Gagnon, F., & Thibodeau, N. (1999). Specificity of Generalized Anxiety Disorder symptoms and processes. Behavior Therapy, 30, 197-207]. The goal of the present study was to clarify the relationship between this cognitive process and worry by experimentally manipulating intolerance of uncertainty. A gambling procedure was used to increase intolerance of uncertainty in one group (N = 21) and to decrease intolerance of uncertainty in another group (N = 21). The results indicate that participants whose level of intolerance of uncertainty was increased showed a higher level of worry, compared to participants whose level of intolerance of uncertainty was decreased. These results provide some initial clarifications as to the causal nature of the link between intolerance of uncertainty and worry. These results are coherent with our theoretical model of worry and GAD (Dugas et al., 1998), which stipulates that intolerance of uncertainty plays a key role in the acquisition and maintenance of excessive worry.

Characterization of the Ly49I promoter.

Fourteen potential Ly49 genes have been identified in the C57B1/6 mouse strain, and cDNAs containing a complete coding region have been isolated for 10 members of this gene family. Ly49 proteins are primarily expressed in natural killer (NK) cells. Although the sequence of the Ly49a promoter region has been published, no study of the cell-specific activity of the promoter has been reported. A 12-kb genomic fragment of the Ly49I gene was isolated and characterized by DNA sequencing. Approximately 5 kb of DNA sequence upstream of the first Ly49I exon was determined and this region was used to perform promoter analysis using luciferase reporter plasmid constructs. A core promoter was identified that was preferentially transcribed in a Ly49-expressing cell line, EL-4. Electrophoretic mobility shift assays using oligonucleotide probes from the core Ly49i promoter and comparable regions from the Ly49a promoter demonstrated the importance of TATA-related elements in generating EL-4 and NK cell-specific DNA/protein complexes.

Cloning and characterization of a novel activating Ly49 closely related to Ly49A.

The majority of the known Ly49 family members have been isolated from either C57BL/6 (B6) or BALB/c mice. Interestingly, the anti-Ly49 Ab reactivities observed in 129/J mice are different from those of B6 mice. Furthermore, immunoprecipitation of 129/J NK cell lysates with YE1/32 and YE1/48, Abs specific for the inhibitory Ly49A in B6, resulted in detection of the activation-associated DAP12 molecule. These results indicated a need for a more detailed study of this strain. Therefore, a cloning strategy was devised to isolate Ly49 cDNAs from 129/J mice. An immunoreceptor tyrosine-based inhibitory motif-containing, Ly49D-related clone was discovered that we have named Ly49O, and one immunoreceptor tyrosine-based inhibitory motif-lacking, Ly49A-related clone was discovered that we have named Ly49P. No anti-Ly49 mAb reacted with Ly49O, whereas the molecule encoded by the Ly49P cDNA was found to react with YE1/32 and YE1/48. Ly49P was found to associate with mouse DAP12, and Ab-mediated cross-linking of Ly49P resulted in mouse DAP12 phosphorylation and Ca2+ mobilization, indicating that Ly49P is a competent activation receptor. Ly49P, therefore, represents a novel member of the Ly49 activating receptor subfamily.

Implementing a multiple-isolator unit for centralized preparation of cytotoxic drugs in a cancer center pharmacy.

Due to numerous reasons: assuring safety (technicians, patients, nurses, environment), preventing medication errors, cost, maintaining pharmaceutical quality, rules and regulations, it was decided to create a dedicated room within the pharmacy for the preparation of intravenous cytotoxic drugs. After a preliminary study, the following choices were made: isolator unit instead of a vertical laminar air-flow hood, rigid surface instead of flexible film, a multiple-unit structure (one half suit unit for storage and one isolator unit for passthrough, two preparation units, each with four double-gloving portals) instead of a single-unit structure. After the equipment was installed, the physical and microbiological processes were validated and a medical/pharmaceutical catalog of protocols was created. Then the technicians were trained and the standard operating procedures were written. Updated every six months, they describe the general organization, gaseous sterilization of the isolators, the drugs and the medical devices, prescription analysis and circuit, preparation of nominative forms and labels, double checking the preparations delivery, cleaning and maintenance, documentation and reports, waste disposal, safety and protection and instructions for emergency. The pharmacists, pharmacy staff, physicians and nurses were all included in a work group responsible for the isolator unit project. The unit was opened in January 1997.