Here we describe the Genome Variation Format (GVF) and the 10Gen dataset. GVF, an extension of Generic Feature Format version 3 (GFF3), is a simple tab-delimited format for DNA variant files, which uses Sequence Ontology to describe genome variation data. The 10Gen dataset, ten human genomes in GVF format, is freely available for community analysis from the Sequence Ontology website and from an Amazon elastic block storage (EBS) snapshot for use in Amazon's EC2 cloud computing environment. With the advent of personalized genomics we have seen the first examples of fully sequenced individuals -9. Now, GVF  is an ex- an ontology developed by the Gene Ontology Consortium [While there are richer ways of representing genomic features using XML (Extensible Markup Language) and relational database schemas, simple text-based, tab-delimited files have persisted in bioinformatics because they balance human with computer readability. Since its adoption as the basic exchange format, two aspects of GFF3 have emerged as essential for success. First, it must be simple for software to produce and parse; second, its contents need to be typed using terms drawn from an ontology. The first aspect means that humans can easily read and edit files with a text editor and perform simple analyses with command-line software tools. The second aspect not only constrains different database curators to use the same terminologies, but also, because of the formal structure of the ontology, allows automated reasoning on the contents of such a file. It therefore prevents ambiguities and conflicting terminologies. GVF builds upon these strengths of GFF3, adopting GFF3's simple, tab-delimited format; and like GFF3, the contents of GVF files are described using the Sequence Ontology (SO) nsortium  to descrnsortium ,20. Usinnsortium , and proBelow we describe the GVF standard and the various additions we have made to GFF3 and SO to support it. We also briefly describe the conversion of the first ten publicly available personal genomes into GVF format. These GVF files are available for download and for cloud computation. We will refer to these data as the 10Gen dataset. This is provided as a service to the biomedical community as a reference dataset for whole genome comparative analyses and software development. This dataset will hopefully foster the development of new tools for the analyses of personal genome sequences.We have extended both the GFF3 specification and SO to allow the rigorous description of sequence variations with respect to a reference genome. The first eight columns of a GFF3 file specify the type and source of a feature, its location on a reference sequence, and optionally a score, strand and phase. These columns of data are incorporated into GVF unchanged. The GFF3 format additionally provides the option to append attributes to a sequence feature using tag-value pairs in the ninth column and it is here that GVF specifies additional structure to annotate sequence alteration specific data Table . EffectiThe SO has been extended in order to describe both the nature of the observed variants and the effects that the variants might have. SO is part of the Open Biological and Biomedical Ontologies (OBO) Library , and folFigure GVF provides nine new pragmas to describe the reference sequence and the methods used to call variants. These pragmas are described in detail in Table Each of the rows in a GVF file describes a single variant from an individual or population. Each such variant is typed using the SO terms that can describe SNVs, any size of nucleotide insertion or deletion, copy number variations, large structural variations or any of the 38 terms currently related to sequence alterations in SO. In the case of a seemingly complex variation, such as an SNV located within a translocation, each sequence alteration is annotated relative to its location on the reference genome, on a separate line in the file.The most flexible part of a feature description in GFF3 is the ninth column, where attributes of a feature are given as tag-value pairs Table . It is hWhile a great deal of personal genome variation data today comes from next generation sequencing technologies, the GVF standard can also be used to describe variant data from any source creating DNA variation data with nucleotide resolution, including genotyping DNA microarrays, comparative genomic hybridization (CGH) arrays, and others.Because GVF is a fully compliant extension of GFF3, GVF files provide a basis for exploration and analysis of personal genome sequences with the widely used Bioperl , and GMODrosophila melanogaster) [et al. [Gold standards and reference datasets are invaluable for software development, testing and for benchmarking the performance of algorithms and tool sets. Classic examples in genomics include the CASP (Competitive Assessment of Protein fold recognition) workshop and its datasets for protein structure comparisons ,33, the ogaster) , EGASP (ogaster) ,36, and ogaster)  datasets [et al.  gene exp- and impact upon - other features annotated on a genome.To fulfill the promise of personal whole genome sequencing it will be critical to compare individual genomes to the reference genome and to one another. One lesson learned from comparative genomics analyses -34,37 isRigorously grounding GVF upon the GFF3 specification has many other benefits as well. Because both file formats are typed using the SO, GFF3 and GVF files can be used together in a synergistic fashion. Moreover, because GVF is a formal extension of the GFF3 standard, existing parsers, visualization tools and validation software, such as those developed by the GMOD project  to operaIn order to jumpstart such analyses, we have also manufactured a reference dataset of variants from ten personal genomes, the 10Gen dataset. These genomes represent a diverse assortment of ethnicities, and were produced using a variety of sequencing platforms. Our hope is that the 10Gen dataset will be used as a benchmark for personal genomics software development, following in the footsteps of other successful benchmark datasets, such as those used by CASP ,33 for pUsing OBO-Edit  the SO wThe variant files from the ten genomes were downloaded from web sites indicated in the references listed in Table The GVF standard can also be used to describe genotyping DNA microarray-based variant calls. This flexibility means that a single parser can process variant files from both sequencing and DNA genotyping microarray experiments; moreover, because these fields are attributes of the variant, not the file, a single GVF file can contain variants from heterogeneous sets of sequencing and microarray platforms.The 10Gen dataset is available for download . Each vaDrosophila melanogaster; GFF: General Feature Format; GFF3: Generic Feature Format version 3; GMOD: Generic Model Organism Database; GVF: Genome Variation Format; NCBI: National Center for Biotechnology Information; NGASP: Nematode Genome Annotation Assessment Project; OBO: Open Biological and Biomedical Ontologies; SNP: single nucleotide polymorphism; SNV: single nucleotide variation; SO: Sequence Ontology; VCF: Variant Call Format.CASP: Competitive Assessment of Protein fold recognition; EGASP: ENCODE Genome Annotation Assessment Project; GASP: Genome Annotation Assessment in MGR, MY and KE conceived the project. BM, CB, FS, LS, KE developed technical aspects. BM maintains the GVF specification and data repository. All authors contributed intellectually to the development of the project. MGR, MY and KE wrote the manuscript with input from the authors.


Discussions of health care reform emphasize the need for coordinated care, and evidence supports the effectiveness of medical home and integrated delivery system models. However, mental health often is left out of the discussion. Early intervention approaches for children and adolescents in primary care are important given the increased rates of detection of mental illness in youth. Most adults also receive treatment for mental illness from nonspecialists, underscoring the role for mental health in medical home models. Flexible models for coordinated care are needed for people with serious mental illness, who have high rates of comorbid medical problems. Programs implemented in the New York State public mental health system are examples of efforts to better coordinate medical and mental health services. Home is the place where, when you have to go there, they have to take you in.The Death of the Hired ManRobert Frost, medical home, large integrated care systems, such as the Mayo Clinic or Kaiser Permanente  children and adolescents, for whom early detection and treatment of mental illness is critical; 2) the general adult population, which receives the bulk of its mental health care in medical settings; and 3) people with serious mental illnesses, who increasingly receive both their mental and primary care in mental health settings.The medical home model originated several decades ago as an approach to coordinating services for children with special health care needs . The modA review of 30 studies of the medical home model in pediatric care indicated that adoption of medical home principles is associated with better health status, better access to care, and improved family functioning for children and adolescents . StudiesTo be viable, medical home and integrated care models need to demonstrate cost-effectiveness. The most compelling data come from North Carolina's Community Care model, which estimated Medicaid savings of $50 million to $260 million per year in 2003 and 2004 following implementation of care networks that incorporate medical home principles . Althougpoint of care. In the medical home model, a primary care physician takes responsibility for coordinating services provided by a team of clinicians. For most adults with mental illness, the point of care would be an internist or family practitioner. People with serious mental illness, however, typically receive most of their care from a mental health clinic. For this population, the successful medical home approach requires a more flexible notion of point of care. Psychiatrists treating people with serious mental illness should monitor basic medical conditions and communicate with primary care practitioners, who provide guidance and specific treatment recommendations , including diagnostic tools, treatment algorithms, and other resources. AAP also released a comprehensive report outlining mental health competencies for pediatric primary care (The American Academy of Pediatrics (AAP) has recognized the prevalence of child mental illnesses, their relationship to adverse early childhood experiences , the undary care . In conjary care . These nary care .As with children, most adults with a mental illness are seen in primary care, not by mental health specialists. Among adults who received care for a past-year episode of major depressive disorder, approximately 60% saw a general practitioner or family doctor, whereas approximately 30% saw a psychiatrist or psychotherapist . AlthougModest improvements in primary care mental health practices increase detection and evidence-based treatment of common mental illnesses for adults  ,41. The Despite these improvements in treating common mental illnesses, improving mental health care in primary care remains challenging. Many patients treated for depression in primary care receive an inadequate trial of medication (insufficient dosage or duration of treatment or both) . In addicollaborative care include embedding mental health professionals  in the primary care setting with accessible psychiatric consultation, screening for mental illnesses, establishing clear treatment guidelines, and measuring the patient's condition periodically. Collaborative care roughly doubles positive depression treatment outcomes; in one study, 45% of patients randomly assigned to a collaborative care intervention had a 50% or higher reduction in depressive symptoms from baseline compared with 19% of usual care participants . Agencies should embed or collocate mental health staff in primary care settings to assist with screening, counseling, and care monitoring and coordination. Likewise, primary care staff should be collocated in mental health agencies to treat or manage low-complexity health problems and coordinate care for complex cases.Provider agencies need to create readily accessible  consultation in which psychiatrists are available to pediatricians, obstetricians and gynecologists, and primary care physicians. They must also continually measure parameters of health or mental health functioning by using meaningful and practical measures of blood pressure, body mass index, smoking status, and depression. Finally, agencies should use a single clinical record  except when specialty care considerations require a distinct record.Whereas the research projects on collaborative care are well known, there are also many home-grown examples of what can be done. Several projects are under way through the New York State Office of Mental Health (NYS OMH) and the New York City Department of Health and Mental Hygiene (NYC DOHMH), and these agencies are not unique in their efforts.In early 2009, all 66 NYS OMH-operated mental health clinics, which had 15,000 adult outpatients, began systematically collecting 3 health indicators: blood pressure, body mass index, and smoking status. Within months, information had been collected on approximately 50% of adult outpatients, with a goal of 100% within a year . Collecting this information is premised on the belief that what gets measured gets managed, and is meant to promote the expansion of wellness programs and primary care collaborations in these mental health clinics.Various NYS OMH and nonprofit community mental health agencies have established medical clinics on-site at their mental health centers. Some are collocated, though remaining separate entities; others are operated by the mental health clinic. The New York State Department of Health has funded 6 demonstration projects that identify high-need people with physical and mental illnesses whose integrated care will be the responsibility of accountable mental health agencies .In 2005, the NYC DOHMH began a citywide initiative to implement depression screening and specified care management in 100% of New York City's primary care practices. This work continues and is reported elsewhere ,48.The medical home concept is a centerpiece of health care reform in this country. The goals for a medical home are that it be accessible, comprehensive, coordinated, culturally responsive, person-centered, and compassionate. It would be an accountable entity where patients and families feel that their interests are primary and attended to by caring clinicians. These goals cannot be achieved if there is no room for mental health in the medical home."There is no health without mental health," said Dr David Satcher in his first Surgeon General's report on mental health . The int


Bacillus anthracis). These data demonstrate increased volume of submissions to a public health laboratory resulting from fear of bioterrorism.After the human anthrax cases and exposures in 2001, the Illinois Department of Public Health received an increasing number of environmental and human samples (1,496 environmental submissions, all negative for  Bacillus anthracis in the environment, to gain insight into the frequency of true versus perceived bioterrorist events, and to observe and respond to the impact of widely publicized terrorism alerts.After the terrorist attacks of September 11, 2001, and the discovery of human anthrax cases and exposures several weeks later on the East Coast, Illinois Department of Public Health’s (IDPH) Division of Laboratories received environmental and human samples for analysis as part of suspected bioterrorism investigations. The large number of samples provided an opportunity to learn about the possible presence of Submissions of powder and nonpowder environmental samples and human blood or tissue specimens submitted from October 8 through December 31, 2001, were reviewed. Before samples were accepted by IDPH Division of Laboratories, incidents involving environmental samples had to be reviewed by Federal Bureau of Investigations (FBI) agents who determined if a potential bioterrorism threat was credible. The IDPH laboratories in Chicago and Springfield processed all samples submitted through law enforcement authorities in this manner. The Chicago laboratory received samples primarily from northern Illinois (north of Interstate 80), while the Springfield laboratory received samples from central and southern Illinois. The Chicago laboratory followed all required guidelines for a biosafety level 3 laboratory  in Chicago and 303 (20.3%) in Springfield. An additional 40 human specimens were processed, 28 (70%) in Chicago and 12 (30%) in Springfield. Chicago sample submissions rose steadily after the first week of October and peaked during the week of October 29 through November 4, with the largest number of submissions processed on November 7 (range 0–64 submissions per day) . An addiBacillus cereus  and nonhemolytic staphylococci  (B. megaterium (four), B. thuringiensis (one), and nonanthracis Bacillus species (three) were identified. Twenty specimens (8 in Chicago and 12 in Springfield) were processed by using PCR. All of these results were negative for B. anthracis.Among both the powders and nonpowders processed at the Chicago laboratory, the most frequently isolated organisms were ctively) . Among tA review of the source and circumstances related to 57 nonhuman samples submitted to the Chicago laboratory on November 7, 2001 (the date with the highest number of submissions), indicated that most of the items were mail items , but powders (7 submissions) and unspecified “suspicious substance” (13 items) also were identified. Such items came from at least six counties and involved 13 local police departments, the Illinois State Police, two fire departments, and the FBI. In addition to submissions from private citizens, submissions came from two universities, a post office, a private business, and a consulate. A high level of anxiety among the public was likely responsible for the otherwise unsuspicious items submitted, including dairy creamer, powder from donuts, a backpack, a telephone, a frozen dinner, a computer keyboard, and a letter from a married man’s lover that was intercepted by his wife and submitted unopened as a suspicious mail item.B. anthracis  could not rule out nthracis . These iB. anthracis among the nearly 1,500 submissions to IDPH for testing. Other state health departments also received large numbers of submissions: Michigan and Oklahoma received at least 228 and 762 submissions, respectively . However, such meetings also competed with laboratory time, which was needed to process the increasing load of submissions.Among the more positive aspects of the high volume of submissions was enhanced cooperation with law enforcement officials, especially the FBI. Such officials maintained a temporary office in the Chicago laboratory facility, where submissions could be received at all hours, prioritized, and then delivered individually or batched for submission to the laboratory (which did not have a night shift of workers) during working hours.Bacillus isolates to determine what species may be expected during such an event, and to anticipate what personnel and changes in protocols might be needed during future bioterrorist events. This experience also highlighted the importance of regular training and updating of laboratory staff on procedures relevant to bioterrorism agent evaluation.Because health department laboratories have previously had only limited and sporadic experience with the test methods involved in urgent microbiologic evaluation for anthrax, the experience with these submissions provided an opportunity to polish critical laboratory skills, to utilize infrequently used or not previously available equipment , to speciate nonanthracis We recommend continued close communication and collaboration between public health and law enforcement officials, which include developing flexible criteria on specimen submission guidelines for public laboratories. Development of standardized forms for information collection related to suspicious substance submissions may be useful in evaluating the epidemiology of any future bioterrorism events. We also recommend regular training of laboratory staff on procedures relevant to bioterrorism agent evaluation, including cross-training of selected staff not usually involved in the handling of suspected bioterrorism agents as part of a surge capacity plan. Such a plan should also include a mechanism for handling submissions after regular work hours because of the high profile and expectations of turnaround time that submissions for bioterrorism agent evaluation usually have. Finally, enhancing communication between the many agencies involved remains both a challenge and an opportunity for future efforts to combat these kinds of events.


Monoclonal antibodies to proteins encoded by the ras, myb, myc, erb-B, src and PDGF-2 genes were tested for reactivity with normal rat liver, livers from rats fed with 0.06% 2-acetylaminofluorene (AAF), and premalignant lesions and primary liver tumours from rats given AAF alone or a combined treatment with diethylnitrosamine and AAF. Radioimmunoassays were performed with plasma membrane fractions and total soluble subcellular extracts of the tissues, and immunoperoxidase staining was carried out on frozen tissue sections. All of the antibodies were positive in radioimmunoassays, some more strongly than others, and each antibody bound equally to extracts of different kinds of tissue. Immunohistology revealed significant staining of normal liver by 5 of the 6 antibodies, and only minor qualitative differences of the staining pattern in some tumours and hyperplastic nodules. It was concluded that these antibodies were not able to discriminate sufficiently well between normal, premalignant and malignant rat liver to be of value in identifying the precursor cells of malignant tumours.


Therefore, we considered additional indirect evidence that would enable a comparison of docetaxel plus prednisolone/prednisone with other chemotherapy regimens and active supportive care. Systematic searching (upto April 2005) identified seven randomised controlled trials. One large well-conducted trial assessed docetaxel plus prednisone vs mitoxantrone plus prednisone; this showed statistically significant improvements with 3-weekly docetaxel in terms of overall survival, quality of life, pain response and PSA decline. Two other chemotherapy regimens that included docetaxel with estramustine also showed improved outcomes in comparison with mitoxantrone plus prednisone. Three trials that compared mitoxantrone plus corticosteroids with corticosteroids alone were identified and their results for overall survival combined, which showed very little difference between the two groups. The addition of clodronate to mitoxantrone plus prednisone showed no significant differences in comparison with mitoxantrone plus prednisone alone. The evidence suggests that chemotherapy regimens containing 3-weekly docetaxel are superior to mitoxantrone or corticosteroids alone.A systematic review was performed to evaluate the clinical effectiveness of docetaxel in combination with prednisolone  for the treatment of metastatic hormone-refractory prostate cancer. A scoping search identified a trial of docetaxel plus prednisone  Prostate cancer is the most common cancer among men in the UK (excluding nonmelanoma skin cancer), the lifetime risk for being diagnosed with prostate cancer is one in 13 . The majTreatment for mHRPC is palliative. Options include second-line hormonal therapy, chemotherapy and active supportive care, and patients may receive a combination of palliative treatments. Current advice issued by the National Institute for Health and Clinical Excellence (NICE) states that chemotherapy should be considered and trials of chemotherapy supported . The taxThe Centre for Reviews and Dissemination (CRD) and the Centre for Health Economics were commissioned to conduct a systematic review on behalf of NICE of the clinical and cost-effectiveness of docetaxel for the treatment of mHRPC. This paper presents the systematic review of the effectiveness evidence. A full technical report is available, which also presents the results of the economic evaluation that was conducted alongside the systematic review  or placebo for the treatment of mHRPC.The objective of the systematic review was to evaluate the clinical effectiveness of docetaxel in combination with prednisolone vs mitoxantrone  plus prednisone , the Cochrane Database of Systematic Reviews (CDSR), National Research Register (NRR), Health Technology Assessment Database (HTA), NHS Economic Evaluation Database (NHS EED), Database of Abstracts of Reviews of Effects (DARE), CINAHL, Health Management Information Consortium (HMIC), ISI Science and Technology Proceedings, Social Science Citation Index, Index to Theses, SIGLE, Inside Conferences, BIOSIS Previews, Current Controlled Trials, ClinicalTrials.gov, International Cancer Research Portfolio (ICRP), National Cancer Institute Clinical Trials PDQ, American Society of Clinical Oncology) were searched from inception to April 2005. Index and free text terms were used to search for prostate cancer and these terms were combined with index and free text terms for docetaxel (including generic and trade names). Separate searches were performed to combine the prostate cancer terms with index and free text terms for mitoxantrone. Where possible, subject index terms were used to exclude animal studies. No language, date or study-type restrictions were applied. The reference lists of included and background papers were also searched for additional relevant studies.Published and unpublished randomised controlled trials (RCTs) that compared docetaxel in combination with prednisolone/prednisone (as per its licensed indication) with any chemotherapy regimen or active supportive care or placebo in men with mHRPC were included. Randomised controlled trials that assessed mitoxantrone in combination with a corticosteroid  compared with any chemotherapy regimen or active supportive care or placebo in men with mHRPC were also eligible for inclusion. Data on the following outcomes were included: overall survival, progression-free survival, response rate , PSA decline, adverse effects of treatment, pain and health-related quality of life.Studies that were reported in abstract form only, where no further information was available, and foreign language papers were excluded. Where multiple publications of the same study were identified, data were extracted and reported as a single study. The quality of RCTs was assessed according to criteria based on CRD Report No. 4 . Each stA total of 1065 titles and abstracts were screened for inclusion in the review and 267 records were ordered as full papers. Seven RCTs were identified that met our inclusion criteria. The process of study selection is shown in Of the seven RCTs included, three used docetaxel compared with mitoxantrone plus prednisone   . Two of these compared mitoxantrone plus prednisone with prednisone (5 mg twice daily) ). This result was exactly the same using fixed effect and random effects approaches. Other outcomes could not be pooled because they were measured differently in the three RCTs. However, in the two studies that measured health-related quality of life and pain response, the mitoxantrone groups had statistically significant improvements compared with the corticosteroid groups for several of the quality of life and pain items assessed    and mitoboplatin et al.The results of the only identified RCT to assess the intervention under consideration (docetaxel plus prednisone) showed statistically significant improvements compared with mitoxantrone plus prednisone in terms of overall survival, quality of life, pain response and PSA decline (when given in 3-weekly doses). The improved outcomes for docetaxel plus prednisone were associated with more grade 3–4 adverse events; however, this had no detrimental effect on quality of life, which was significantly improved. There were statistically significant improvements in quality of life and PSA decline for patients receiving weekly docetaxel, however, survival and pain response were not statistically significantly improved. Two other chemotherapy regimens that included docetaxel: docetaxel plus estramustine and docetaxel plus prednisone plus estramustine, also showed improved outcomes in comparison with mitoxantrone plus prednisone in terms of overall survival, progression-free survival, response rate and PSA decline. However, docetaxel in combination with estramustine was associated with more adverse events. It should be noted that these trials included patients with a Karnofsky performance status score of at least 60%, or Eastern Cooperative Oncology Group or Southwest Oncology Group performance status score of 0–2; therefore, the results can only be generalised to patients with a similar performance status.Three trials that compared mitoxantrone plus a corticosteroid with a corticosteroid alone were identified and their results for overall survival were combined, which showed very little difference between the two groups. The addition of clodronate to mitoxantrone plus prednisone showed no significant differences in comparison with mitoxantrone plus prednisone alone.The review was limited by the lack of trials assessing the intervention under consideration: docetaxel plus prednisone. As docetaxel plus prednisone has only been directly compared with mitoxantrone plus prednisone, we considered additional evidence that would enable a comparison of docetaxel plus prednisone with other chemotherapy-based treatments and active supportive care. The only form of active supportive care for which evidence was available was corticosteroids. Therefore, no conclusions can be drawn regarding the comparative effectiveness of docetaxel to other types of active supportive care.The small number of studies identified prevented the assessment of publication bias using funnel plots or the Egger test . Careful patient selection by the oncologist therefore remains paramount.The limited evidence suggests that 3-weekly docetaxel plus prednisone is superior to mitoxantrone plus prednisone in terms of overall survival, quality of life, pain and PSA decline, and that mitoxantrone plus a corticosteroid does not improve survival compared with a corticosteroid alone. At the time of this assessment, there were ongoing trials of docetaxel as the standard arm in combination with other therapies.


Effects of heating and fasting, both alone and associated, have been studied in normal and Sarcoma 180 bearing mice. Heating reduced body weight and tumour mass and increased body temperature. Fasting reduced body weight, while tumour mass and body temperature were slightly affected. By associating heating and fasting a more marked decrease of body weight was produced than by each of the two factors involved, while effects on body temperature and on tumour mass were unchanged with respect to heating alone. 6-mercaptopurine was similar to heating in reducing body weight and tumour mass.


Castanea dentata) was devastated by an exotic pathogen in the beginning of the twentieth century. This chestnut blight is caused by Cryphonectria parasitica, a fungus that infects stem tissues and kills the trees by girdling them. Because of the great economic and ecological value of this species, significant efforts have been made over the century to combat this disease, but it wasn't until recently that a focused genomics approach was initiated. Prior to the Genomic Tool Development for the Fagaceae project, genomic resources available in public databases for this species were limited to a few hundred ESTs. To identify genes involved in resistance to C. parasitica, we have sequenced the transcriptome from fungal infected and healthy stem tissues collected from blight-sensitive American chestnut and blight-resistant Chinese chestnut (Castanea mollissima) trees using ultra high throughput pyrosequencing.American chestnut (C. mollissima and C. dentata respectively.We produced over a million 454 reads, totaling over 250 million bp, from which we generated 40,039 and 28,890 unigenes in total from In silico expression analyses showed that many of the stress response unigenes were expressed more in canker tissues versus healthy stem tissues in both American and Chinese chestnut. Comparative analysis also identified genes belonging to different pathways of plant defense against biotic stresses that are differentially expressed in either American or Chinese chestnut canker tissues.The functions of the unigenes, from GO annotation, cover a diverse set of molecular functions and biological processes, among which we identified a large number of genes associated with resistance to stresses and response to biotic stimuli. Cryphonectria parasitica between American chestnut and Chinese chestnut.Our study resulted in the identification of a large set of cDNA unigenes from American chestnut and Chinese chestnut. The ESTs and unigenes from this study constitute an important resource to the scientific community interested in the discovery of genes involved in various biological processes in Chestnut and other species. The identification of many defense-related genes differentially expressed in canker vs. healthy stem in chestnuts provides many new candidate genes for developing resistance to the chestnut blight and for studying pathways involved in responses of trees to necrotrophic pathogens. We also identified several candidate genes that may underline the difference in resistance to  Castanea), members of the family Fagaceae, naturally occur throughout deciduous forests of eastern North America, Europe, and Asia [Castanea mollissima), Japanese chestnut (Castanea crenata), European Chestnut (Castanea sativa) and American chestnut (Castanea dentata).The chestnuts  was also devastated by introduction of C. parasitica [Castanea exhibit very different susceptibilities to Cryphonectria infection. Asian chestnuts, the vector for the spread of Cryphonectria westward, range from somewhat susceptible to nearly immune to infection [Cryphonectria. Slow growing cankers are often visible on Chinese and Japanese chestnut trees although growth and yield of the trees are not substantially reduced. European chestnut is able to tolerate infection slightly more than American chestnut, which has little or no natural resistance to Cryphonectria infection [Although to a lesser extent, European chestnut . The average length of the reads was 101 nucleotides (nt)  and 213 (~1.5%) of the American and Chinese chestnut unigenes, respectively.The American Chestnut Canker cDNA library was constructed from a pool of RNA isolated from canker tissues of several individuals of one genotype (BA69). The Chinese chestnut cDNA library was also prepared from RNA extracted from several individuals of a single genotype (Nanking). One plate of sequencing was conducted with each library using the GS20 model of the 454 system. A total of 129,508 and 235,635 reads were generated from American and Chinese chestnut canker transcriptomes respectively, in the GS20 runs , with an average read length of ~246 nt  with an average read length of 247.9 bases . A quarter plate of sequencing was conducted for each American chestnut healthy stem library, while a three quarter plate worth of sequencing were conducted for the Chinese chestnut Nanking and Mahogany libraries Table . SequencArabidopsis proteome, a function could be assigned to 83,292 (26%) and 20,391 (28%) of the 454 reads from American and Chinese chestnut respectively. These percentages are lower than those obtained by BLASTx alignments to the Populus proteome (39% and 46% for American and Chinese chestnut respectively). However, most of the reads  with best hits to the Populus proteome are for Populus genes that are annotated as having no known function. GO ontology analysis based on the Arabidopsis proteome showed that the distributions of gene functions for cDNA sequences from American and Chinese chestnut cankers are similar   classificinnamoyl-CoA reductase (CCR), 4-coumarate–CoA ligase, hydrolase, kinases, phosphatases, translation factor, ATPases, pathogen responsive alpha-dioxygenase, etc . While a fraction of the remaining sequences may correspond to 3' or 5' untranslated regions, non coding RNAs, or short sequences not containing a known protein domain, a large number may correspond to potential Chestnut-specific genes. A similar situation was found when analyzing the transcription of Eschscholzia californica, Persea americana, and Aristolochia fimbriata [Before this project, only a few hundred chestnut sequences had been deposited in the EST database (dbEST) at NCBI. The data presented here represent the first large effort by the Fagaceae Genomic Tools Development project to generate cDNA resources and analyze the transcriptomes of American and Chinese chestnut. These resources are public and the sequences can be accessed in a searchable database at the project website , or as rimbriata  and  of coverage by 454 gene tagging, makes this technique a good tool for quantifying the expression level of sets of genes involved in various developmental stages or physiological conditions. cDNA sequences generated from both species cover various biological processes and molecular functions indicating that 454 sequencing constitutes a powerful tool for sequencing the transcriptome of non model species. These results confirm that pyrosequencing constitutes a powerful tool for transcriptome characterization and gene discovery.By taking into consideration only the sequences that have homologies in the Cryphonectria) mycelium within the canker and function by weakening the plant cell wall to facilitate fungal entry. However, analysis of these hydrolase sequences showed that they are more similar to other plant hydrolase genes than to fungal hydrolase sequences. This suggests that these hydrolase proteins are of plant origin and function either by strengthening cell walls against pathogen entry or in the programmed cell death response of the cells at the fungal infection site in the chestnut stem. A second functional category well represented is kinase activity. Such genes are involved in signaling in pathogen infection and play a key role in plant defense response. A third functional category observed in the chestnut transcriptomes is represented by transcription factors or genes associated with RNA or protein binding. Such genes may modulate the expression of resistance genes in response to the pathogen infection.GO annotation analyses showed that, overall, canker tissues from both species present a similar transcriptome. Gene function categories associated with metabolic process are highly represented in both transcriptomes. The category represented the most is composed of genes associated with various metabolic processes as previously described in other systems such as cassava . The secABC transporter, C2-domain-containing gene, methylenetetrahydrofolate reductase, elongation factor-1 alpha, and peroxidase. Such genes are involved in controlling the extent of the cell death in the defense response [Pseudomonas syringae [omega-3 fatty acid desaturase, suppressor of fatty acid desaturase deficiency (SFD1 and SFD2), Ras-related GTP-binding which are required for systemic resistance [ATPase was found to be over-expressed in American and Chinese chestnut. This gene is required for the attenuation of the hypersensitive response [mitogen activated protein. This protein kinase activates both local resistance and basal resistance [Acetyl co-enzyme A carboxyltransferase (CAC3), SNF, and several transcription factors such as WRKY, Zinc finger, Myb, etc were identified. Myb genes are involved in regulation of disease resistance genes [PAL2, a key enzyme in phenylpropanoid and lignin biosynthesis [WRKY transcription factors have been shown to fine tune the response of plants to challenge with pathogens [SNF genes interact with other genes, such as SnRK1, which regulate glucose metabolism, cell defense and other cellular processes.Among genes that were found to be differentially expressed in American or Chinese chestnut or both, several are known to be involved in various processes of plant defense against pathogens such as cell death related to hypersensitivity response, construction of a physical barrier to block the pathogen progression, as well as systemic resistance. Among genes involved in hypersensitivity cell death, we found response -34. Pleiresponse -37. The response -42 showesyringae . We alsosyringae . Chestnusistance ,45. ATParesponse . Among gsistance ,46,47. Ice genes -50; theyynthesis . WRKY trathogens . SNF genCryphonectria parasitica infection by activating both local and systemic resistance responses. The trees try first to block the progression of the pathogen by increasing the expression of hydrolases, lignin synthesis, and cell death. The infection is also sensed by mitogen kinases, which activate other transcriptions factors such as AP2, Myb, and WRKY, which in turn induce the expression of genes from the phenylpropanoid, jasmonic acid, oligochitosan, and other pathways that are involved in resistance to pathogens [Overall, this study allowed us to conclude that chestnut trees respond to athogens ,54.Cryphonectria parasitica, (iii) Identify potential pathways involved in chestnut resistance to the Cryphonectria parasitica, and (iv) Identify several candidate genes for resistance to necrotrophic fungal pathogens in trees.In conclusion, this study allowed us to (i) Obtain over 28,000 and 40,000 unigenes from American and Chinese chestnut, (ii) Compare the transcriptomes of American and Chinese chestnut following infection by Cryphonectria strain EP155 as described by Hebard and collaborators [Healthy cambial tissue was collected from the American chestnut genotypes 'Watertown' and "Wisniewski' growing at the Connecticut Agricultural Experiment Station, Lockwood Farm, Hamden CT. Canker tissue was collected from the American chestnut genotype BA69 growing at The American Chestnut Foundation Meadowview Research Farm, Meadowview VA. Healthy cambial tissue was collected from the Chinese chestnut blight resistant genotypes 'Nanking' and 'Mahogany' growing at The American Chestnut Foundation Meadowview Research Farm, Meadowview VA. Canker tissues were collected from Chinese chestnut genotype 'Nanking' growing at Meadowview Research Farm. To create cankers, the stems of Chestnut trees were inoculated with the hypervirulent 2, again extracted with chloroform and precipitated with ethanol. The resulting RNA pellet was resuspended in 40–100 μl of DEPC-treated water, and the quality was assessed with an Agilent Technologies 2100 Bioanalyzer (Agilent Technologies). Poly(A) RNA was then separated from total RNA using the Poly(A) Purist kit (Ambion) and the quality assessed with an Agilent Technologies 2100 Bioanalyzer (Agilent Technologies). cDNA was synthesized from the mRNA using the Just cDNA kit (Stratagene) using random hexamer primers provided with the kit to obtain better 5' to 3' coverage of transcripts than is possible using Poly(A) priming alone.Total RNA was prepared by the method of Chang and collaborators . Three tThe resulting cDNA was used to construct a 454 library following the supplier's instructions (Roche Diagnostics). The sequencing was conducted at Penn State University using an FLX model 454 DNA sequencer (Roche Diagnostics).SRX001799, SRX001800, SRX001801, SRX001804, RX001805, and SRX001806 (submission SRP000395).454 libraries were constructed as described previously . In summArabidopsis [Populus [Cryphonectria parasitica [Arabidopsis gene identifiers with the strongest BLASTx alignments to the corresponding chestnut 454 reads. Comparison of the distribution of biological processes or molecular function obtained using Go annotation was done using GOstat program [The data from the 454 read sequences were assembled into transcript contigs using Newbler Assembler software (Roche). Reads from each library were assembled separately. The unigene (contigs and remaining unique singletons) sequences were annotated by query against the proteomes of bidopsis ) and Pop[Populus  and the rasitica  using Bl program . Compari program .CAD: cinnamyl alcohol dehydrogenase; CCR: cinnamyl-CoA reductase; cDNA: complementary DNA; EST: expressed sequence tag; GO: Gene Ontology; Mb: megabases; NCBI: National Center for Biotechnology Information; nt: nucleotide; SAGE: Serial Analysis of Gene Expression.AB contributed to extracting RNA and making the 454 libraries, curated and analyzed the data, supervised the work SC and YZ, and wrote the paper. CS and YZ contributed to the bioinformatics analyses. DSD collected tissue samples, prepared RNA, cDNA and 454 libraries, and helped prepare the first draft of the manuscript. KB and WAP contributed to the RNA preparation and discussion of disease response gene candidates. NW manages all aspects of the Genomic Tool Development for the Fagaceae Project. RS is the Principle Investigator of the Genomic Tool Development for the Fagaceae Project and was responsible for oversight, budget, obtaining the funding for the project, and contributing advice at each step of the research. This work was conducted in the laboratory of JC, who initiated the research with American Chestnut Foundation funding, co-directs the 454 sequencing facility at Penn State, and contributed to the development of 454 sequencing protocols, evaluation and discussion of the results, and preparation of the manuscript.Genes more highly expressed in canker tissues than healthy stem tissues of American Chestnut.Click here for fileGenes more highly expressed in canker tissues than in healthy stem tissues of Chinese Chestnut.Click here for fileDisease and Defense Response Genes More Highly Expressed in Infected Tissues of Chinese Chestnut (CC) than in American Chestnut (AC).Click here for fileDisease- and Defense- Response Genes More Highly Expressed in Infected Tissues of American Chestnut (AC) than in Chinese Chestnut (CC).Click here for file


Although cigarette smoking is a principal risk factor for bladder cancer in both men and women, few studies have statistically evaluated whether gender modifies the effect of smoking on bladder cancer risk. We initiated the present case-control study at Roswell Park Cancer Institute in Buffalo, New York, U.S., to provide further data on this important issue. We observed similar risk estimates for men and women with comparable smoking exposures, but did not observe a statistically significant interaction between gender and lifetime smoking exposure. We conclude that cigarette smoking is a major risk factor for bladder cancer in both sexes, but that gender does not modify the effect of smoking on bladder cancer risk. Cigarette smoking is a major independent risk factor for urinary bladder cancer. Many epidemiologic studies conducted in various parts of the world have shown that both male and female cigarette smokers have two to four times the risk of developing bladder cancer compared to nonsmokers. In general, the risk increases as the frequency and duration of smoking levels increase, and risk typically decreases in former smokers compared to current smokers ,2.In a recent case-control study conducted in Los Angeles, California, U.S., Castelao and colleagues  reportedThis study was approved by the Institutional Review Board of Roswell Park Cancer Institute (RPCI) in Buffalo, New York, U.S., and informed consent was properly obtained from all participants. Study subjects included adult patients admitted to RPCI between 1982 and 1998, who volunteered to complete a self-administered epidemiologic questionnaire. The details of this hospital-based data collection system have been previously described . Cases , and the usual number of cigarettes smoked daily during years of regular use (frequency). A summary measure of lifetime smoking exposure (pack-years) was calculated as the product of frequency (packs/day) × duration (years).Unconditional multiple logistic regression was used to calculate odds ratio (OR) and 95% confidence interval (CI) estimates, and to test for the statistical significance of risk trends. OR and 95% CI estimates presented in this report are adjusted solely for age, which was entered as a continuous variable. Further adjustment for potential confounders such as occupational exposures , education, income, and geographic area of residence failed to notably alter point estimates. Gender was evaluated as a potential effect modifier for the association between smoking and the risk of bladder cancer by testing for the statistical significance of an interaction term  in a logistic regression model . All datSubject cigarette smoking histories and the results from the logistic regression analyses are presented in Table The risk of bladder cancer generally increased in both sexes as pack-years, duration, and frequency levels increased. We observed similar risk estimates for males and females with comparable smoking histories, and did not find a statistically significant interaction between lifetime smoking exposure and sex P = 0.76), thereby leading us to conclude that gender was not an effect modifier for the association between smoking and bladder cancer in our case-control data. In the two other case-control studies that have formally tested for the statistical significance of an interaction term , Castelao and colleagues  detected6, therebThe results of this study are consistent with those of other recent case-control studies from the United States -10 and CIt is important to consider that in most populations worldwide, bladder cancer is relatively rare in women. Men have approximately three to four times the risk of developing bladder cancer compared to women, and this sex difference in risk persists even after controlling for differences in smoking and occupational exposures ,2,5,9. TThe authors declare that they have no competing interests.


Musculoskeletal conditions, such as low back pain, are prevalent in the United States. These conditions exact an enormous toll on society, both in terms of their detrimental impact on quality of life and on the costs of treatment and lost productivity. Osteopathic physicians, as common providers of primary care services and spinal manipulation, are ideally positioned to lead future research efforts in this field. The emergence of data and standards relevant to osteopathic manipulative treatment outcomes, refinement of research methodologies to enhance evidence-based medicine, and investments in developing osteopathic research infrastructure are all critical elements in moving this field of research forward. He conc study." . Over 30 study." . CorrespOsteopathic physicians in the United States manage a disproportionately large number of patients with low back pain . In so dData from the National Health Interview Survey (NHIS) in 2007 indicate that 14.3 million adults used complementary and alternative medicine (CAM) therapies for back pain within the past 12 months, making it by far the leading reason for CAM therapy use . Neck paThe volume of patients with low back pain managed by osteopathic primary care physicians and their dual roles as conventional and CAM practitioners suggest that the osteopathic profession is ideally positioned to respond to Deyo's challenge, particularly as it pertains to spinal manipulation for low back pain. The osteopathic profession should take the lead in providing the scientific evidence base to establish the safety, efficacy, and effectiveness of spinal manipulation in treating low back pain and other musculoskeletal conditions. This should involve a comprehensive approach, including basic science research, translational research, clinical trials, and comparative effectiveness studies.Although spinal manipulation has been used in treating low back pain for over a century in the United States, the Agency for Health Care Policy and Research did not issue its seminal clinical practice guideline until 1994 .A review of nonpharmacologic therapies for acute and chronic low back in 2007 reported that the therapies with good evidence of moderate efficacy for subacute or chronic low back pain were cognitive behavioral therapy, exercise, spinal manipulation, and interdisciplinary rehabilitation, whereas only superficial heat was associated with good evidence of efficacy for acute low back pain . The joialternative to conventional medical treatment for low back pain, but not as a complement to conventional medical treatment. The lack of guidelines on complementary spinal manipulation in the United States represents an important scientific knowledge gap because patients often seek conventional and CAM therapies concurrently.Thus, spinal manipulation has been promoted as an In May 2009, the National Institute for Health and Clinical Excellence (NICE) in the United Kingdom issued its clinical guideline on the early management of persistent non-specific low back pain . A goal "...it gives clear, although still preliminary, evidence that OMT is much more than placebo," and recommended that two sufficiently powered trials be conducted to better delineate the magnitude of OMT effects [Most studies of spinal manipulation for low back pain have focused on manipulation provided by chiropractors or physical therapists, not by osteopathic physicians or osteopaths. Although Goldstein challenged the osteopathic profession in 1997 to embrace evidence-based medicine , there h effects .The OSTEOPAThic Health outcomes In Chronic low back pain (OSTEOPATHIC) Trial is a phase III clinical trial using a randomized, double-blind, placebo-controlled, 2 × 2 factorial design to study the efficacy of OMT . UltrasoOver the past few years, new data and standards have emerged to facilitate evidence-based medicine within the realm of OMT. A re-analysis of data originally collected in the North Texas Clinical Trial of OMT for chronic low back pain  was perfHaving standards for minimally important changes enables the implementation of evidence-based medicine, including the determination of such measures as number needed to treat, number needed to harm, and likelihood of help vs. harm as they pertain to OMT . Using sThe spinal manipulation guidelines described above have been driven by "efficacy" studies . Such studies generally include highly selected subjects treated under rigid experimental protocols. However, data are needed to corroborate the benefits of spinal manipulation in real-world settings under less-than-optimal conditions. Such "effectiveness" data, in some ways analogous to pharmaceutical post-marketing surveillance data (phase IV), are most efficiently collected in observational studies. Consequently, new paradigms are needed for extending research on OMT for musculoskeletal conditions to include thousands of patients. One such approach, for example, involves observation of osteopathic physicians providing OMT to their patients without the artificial constraints of a clinical trial protocol.The Osteopathic Research Center (ORC) recently developed a framework for moving to this next step in musculoskeletal research, with a focus on chronic low back pain. The objectives of this ORC initiative include: (1) creating a Consortium for Collaborative Osteopathic Research Development (CONCORD); (2) training a cadre, initially of 24 clinical research fellows, with specific interests in OMT for chronic low back pain; and (3) developing a national practice-based research network (PBRN) using the clinical practices of these fellows. Ideally, the location and clinical practice characteristics of each fellow could be used to configure a geographically representative and population-based PBRN for research in this field. An inception cohort of 2,000 to 3,000 patients with non-specific chronic low back pain could be enrolled over two years, providing data on clinical and laboratory findings. The fellows' training would facilitate fidelity in documenting treatment interventions and measuring outcomes. This framework would provide a foundation for planning and implementing more rigorous studies as "spin-offs," including nested case-control studies, longitudinal studies, and clinical trials.There is a great need for research involving the costs in providing OMT for musculoskeletal conditions . BecauseCost-utility analysis is particularly attractive for studying the effects of OMT provided as a complement to conventional medical treatment for low back pain. This approach measures the incremental costs of OMT needed to achieve an incremental improvement in health, often measured in quality-adjusted life years (QALYs). The costs of providing OMT in an ambulatory setting are often straightforward, and either can be directly measured by the health care provider or estimated using explanation of benefits statements or other third-party billing data. Recent advances in econometric modeling have led to the development of algorithms for converting common outcomes data, such as the Medical Outcomes Study SF-36, into QALYs . Thus, tMusculoskeletal conditions, such as low back pain, are prevalent in the United States. These conditions exact an enormous toll on society, both in terms of their detrimental impact on quality of life and on the costs of treatment and lost productivity. Osteopathic physicians, as common providers of primary care services and spinal manipulation, are ideally positioned to lead future research efforts in this field. The emergence of data and standards relevant to OMT outcomes, refinement of research methodologies to enhance evidence-based medicine, and investments in developing osteopathic research infrastructure are all critical elements in moving this field of research forward.Dr. Licciardone is executive director of The Osteopathic Research Center.


In 1988 South Korea opened its cigarette market to foreign companies under the threat of US trade sanctions. Despite strong social stigma against female smoking in South Korea, and restrictions on tobacco marketing to women and children, smoking rates among young Korean females increased from 1.6% in 1988 to 13% in 1998. Previous analyses describe how Asian countries have been targeted by transnational tobacco companies for new markets, with Asian females offering substantial future growth potential. An understanding of the strategies used by TTCs to increase smoking among Korean females is critical to public health efforts to adopt a stronger gender perspective in implementing the Framework Convention on Tobacco Control.Internal documents of transnational tobacco corporations were systematically searched using keywords focused on the targeting of the female market since market liberalisation in 1988. Industry documents were analysed alongside primary and secondary data on the tobacco industry in South Korea.TTCs have targeted Korean females since the late 1980s, conducting market research to understand consumer preferences, cultural characteristics and social changes affecting women and girls. Brands designed to appeal to females have focused on "slim" and "superslim" cigarettes, "light" and "mild" claims, and marketing which appeals to the growing numbers of young women entering the labour force. Strategies for overcoming legal restrictions on marketing to women and children have included the use of company rather than brand names, retail distribution at venues frequented by females, trademark diversification and sponsorship.Given the high male smoking rates in South Korea, tobacco control efforts have given limited attention to girls and women. The limited data available on female smoking behaviour suggests that, despite legal restrictions and social stigma, smoking among females has increased since market opening, notably within younger age groups. In addition to more detailed trend data, there is an urgent need for the development and implementation of gender-sensitive tobacco control measures. Part of South Korea's accession to the FCTC should include emphasis on measures to address the strategic targeting of Korean females by TTCs. The Kobe Declaration, agreed upon at the WHO International Conference on Tobacco and Health in 1999, draws particular attention to the vulnerability of women and girls to the tobacco epidemic. WHO predicts that female smokers will triple in number (from 200 to 600 million by 2025), and the Declaration concludes " [i]t is urgent that we find comprehensive solutions to the danger of tobacco use and address the epidemic among women and girls" lthough in many countries male incidence of smoking is plateauing or even declining, female incidence appears to be more robust often showing continued positive trends" . Like thOur main postulated explanations for it are the increasing liberation of females throughout the world and the greater quantities of disposable income which they are obtaining. The first of these factors increases the degree of stress placed upon women in their day to day lives and/or makes them more interested in adopting traditionally male habits such as smoking as symbols of their equality. The second factor, personal income, gives them the freedom to indulge in the habit. This move to sexual equality is liable to continue in the future and lead towards an equalisation of male and female incidence of smoking and consumption. Thus, although male consumption may plateau or even decline, female consumption will continue to rise [ to rise .By the early 1990s, these ideas began to be applied to Asian women. As many countries in the region, including South Korea, experienced rapid economic growth, it was anticipated by industry analysts that the changing role of women would offer TTCs an opportunity to remain globally competitive:Long-term international-tobacco outlook: Bullish....We believe that a combination of factors has created a large one-time opportunity for global competitors who can move quickly:...Easing trade restrictions in key Asian markets, notably Japan, Thailand, Korea, and Taiwan, should fuel export growth of 18% per year between 1992 and 1996....With rising social status and participation in the labor force, more Asian women have taken up smoking [ smoking .International Herald Tribune, smoking was once "taboo" for women in many Asian countries, but had become a sign of female emancipation [In South Korea, the potential for TTCs to exploit rapid economic development and social change was immediately recognised. While Korean adult males had among the world's highest rates of smoking prevalence, there was a traditionally strong social stigma against female smoking. The country's significant economic growth from the 1960s, and integration into the global economy , broughtcipation ,50. In mcipation . At thiscipation  for forecipation .TTCs began to focus attention on better understanding the female market in South Korea in the mid-1990s. The desire to increase the number of female smokers grew as smoking prevalence among adult Korean males began to decline. BAT predicted that, as sales volume declined among older adult males, this would be offset to some degree by increasing female smoking . A 1995 The National Health Promotion Law Enforcement Ordinance, adopted in 1989, bans all tobacco advertising, marketing and sponsorship targeted at women and children including both print and broadcast media. Despite this restriction, smoking rates among Korean females has generally increased Table , with evFinesse, based on research showing that "the majority of purchasing and consumption of cigarettes by Korean females is made in coffee shops" [Second, TTCs focused on retail distribution on venues which tended to be frequented by females. Documents describe the sale of female brands in restaurants, coffee shops, "event lunches" , bars, ne shops" .Third, TTCs have used "trademark diversification" (TMD) to circumvent restrictions in order to promote selected brands to the female market. A 1999 BAT document defined TMD, sometimes known as brand stretching, as "the extension of a well-known trademark and its associated brand essence to a product or service unrelated to the one for which the trademark is traditionally associated" . ExampleIn a global environment of ever increasing restriction in the availability of traditional advertising media, parallel communications devices such as sponsorships and trademark diversification now represent the only major alternatives for tobacco marketers in a growing list of markets. Where traditional advertising media are available, their use should be maximized. However, in markets where these media are no longer available or are threatened in the foreseeable future, parallel communications should be seriously considered as part of a brand's marketing mix [ting mix .Capri/Finesse. A 1996 B&W report noted that, despite legal restrictions, KT&G had advertised its brand Simple in numerous magazines aimed at female readers. Strategies included the coupling of cigarettes with bottles of Chanel perfume [In 1989, PM introduced the first consumer pack promotion in Korea for Virginia Slims which included a pocket size address book . In 1990 perfume , and theKent "were very image compatible...a creative natural fit for upscale sociability in a resort setting" [Finally, TTCs used sports sponsorship to target certain age groups within the female market. In 1988, PM International noted that a tennis exhibition for Virginia Slims as a sponsorship would be acceptable, but cautions about associated perceptions noting "we have to be careful that we don't seem to be "targeting" females" . At the setting" .To capture the female market, TTCs undertook extensive research to develop products that would appeal to it. Findings identified female-specific styles and product preferences and specific types of packaging. For example, it was predicted that menthol flavoured cigarettes would increase slightly as a result of the growth in female smoking . It was Cigarette size was also identified as a characteristic that would appeal specifically to females. Based on a survey of Korean smokers aged between 18 and 54 years , PM assessed the importance of different product themes and attributes by gender. Male participants rated thicker circumference cigarettes higher than female participants, and women rated cigarettes with less smoke and no smoke higher than men . Brown &Finesse , "consiampaign" . Marketeampaign"  women,CAPRI/FINESSE is the U.S. International superslim cigarette offering the female smoker a statement of contemporary feminine style and a quality product. CAPRI is a fashion accessory. Target smokers are sophisticated, young adult females 21–35. The total CAPRI/FINESSE proposition projects a top quality image and makes the female smoker feel more like a woman [ a woman .Finesse combined the growing worldwide popularity of "light" cigarettes with distinctive "superslim" dimensions:Capri/Finesse is targeted to the largest group of prospective superslim smokers, women. Prior to the introduction of superslims, brands positioned primarily to women held a small share of the world market...Capri/Finesse can be effective against those brands.Finesse smokers were described as younger (under 30 years of age), well-educated, more likely to be single, and having above average family incomes. As described by BAT, "Smoking Capri/Finesse makes a woman feel good about herself as a woman and allows her to make a style statement" [Capri/Finesse given high levels of support from consumers for a superslim product [atement" . Importaatement" . South K product . Volume  product .Marlboro Lights performed poorly on taste-related attributes [By the late 1990s, BAT noted that improvements in taste and quality were needed to appeal more broadly to women. A 1997 BAT General Consumer Survey (GCS), a large, quantitative study (n = 500), investigated female-specific smoker usage, attitudes, behaviour, and brand images through face-to-face interviewing in places where females commonly smoked. The results indicated that, despite continued social stigma towards female smoking, female smokers were moving away from imported cigarette brands and showing signs of "maturity" . Korean tributes .Virginia Slims, the flagship female brand for the company worldwide. A 1989 PM report, Korea Market Management, recognized that the market for Virginia Slims Light (VSL) was growing at an impressive rate. To ensure continued growth, the company aimed to: (a) keep VSL in line with the trend towards lower tar and perceived product strength; and (b) introduce an ultra low version of the brand. At the same time, PM planned to launch a super slim product that would gain market share from Finesse, recognised as the only imported brand with a high share of female smokers (16%): "Since Finesse has a higher appeal to higher income female, a well-refined image for a Super slim brand with an acceptable product may have a chance to gain smokers from Finesse" [Virginia Slims and Super Slims together had become the second largest import brands behind Japan Tobacco's Mild Seven [In contrast with BAT, PMI targeted the female market with a single brand, a "light" version of Finesse" . By the ld Seven .The historically high rates of smoking prevalence in South Korea, the world's highest at one time among adult males, spurred TTCs to seek market access in the 1980s. While gaining market share among male smokers was the initial aim, this analysis describes how TTCs have targeted girls and women as a promising source of future profits.Tobacco control efforts by the Korean government and public health advocates to date have largely focused on reducing the number of male smokers. These efforts have been reported to have reduced male smoking rates, from 79% 1980) to 44% (2007) [ to 44%  makes clear reference to the need for a gender perspective in strengthening tobacco control measures. As stated in Article 4, gender must be considered across all control policies adopted and implemented by signatories . The finFirst, the limited data available on female smoking prevalence and behaviour in South Korea must be urgently addressed. Data from the Korean National Health and Nutrition Survey Table  suggestsSecond, fuller and more detailed data on female smoking behaviour would support more effective targeting of tobacco control measures. A KASH survey found that the smoking rate by high school girls (17–19 years) rose from 1.8% to 8.1% between 1988 and 1997, a finding supported by other surveys ,84. SmokFinesse was popular among young females during the first half of the 1990s [Esse Menthol (KT&G) and Virginia Superslims  have dominated market share since the late 1990s, all designed to appeal to the female market. So-called "ultra light", "low tar" and "superslim" cigarettes have been particularly effective, suggesting certain brands offer a healthier or safer option, as well as appealing to female concerns about weight gain. Restrictions on the use of such descriptors, alongside public education on the fallacy of such claims, are needed. The creation and promulgation of certain aspirations and values to appeal to Korean females, in general, should be addressed by public health advocates through public disclosure and countermarketing.Third, documents reviewed in this paper support the need for stronger gender-based tobacco control measures that counter the strategic targeting of Korean females by TTCs. Product design associating smoking with body image and female emancipation, familiarly deployed elsewhere , have behe 1990s , while EFinally, the strategic targeting of females in South Korea by TTCs since market opening has occurred despite Article 14 of the National Health Promotion Act of 1989 which is intended to restrict advertising directed at women and children. These findings suggest that these restrictions as currently stipulated have been effectively circumvented and exploited by tobacco companies. In particular, the Act has not prevented the use of indirect marketing tactics, such as brand stretching, sponsorship of events, or the use of descriptors or product design that appeal to females. There is need for comprehensive tobacco control legislation in South Korea, commensurate with commitments under the Framework Convention on Tobacco Control, which bans all forms of tobacco advertising, marketing and promotion.Since the opening of the South Korean tobacco market in the late 1980s, females have been targeted by TTCs as an important source of future market growth and profitability. The rise in smoking rates among females within certain age groups since the late 1980s suggests that these efforts have been successful. The implementation of comprehensive tobacco control measures under the FCTC, from a gender perspective, is urgently needed to protect and promote the health of Korean women and girls.BAT: British American Tobacco; B&W: Brown and Williamson; GATT: General Agreement on Tariffs and Trade; KT&G: Korean Tomorrow & Global; JTI: Japan Tobacco International; PMI: Philip Morris International; TTC: transnational tobacco company; USTR: United States Trade Representative.KL collaborated with the University of California, San Francisco and the Mayo Clinic in the Guildford Archiving Project which created the BAT Document Archive. KL has received funding for tobacco document research from the Rockefeller Foundation, Wellcome Trust, Cancer Research UK and Health Canada.KL, CCa and CCh undertook systematic searching and analysis of internal industry documents. KL and CCa drafted and revised the paper. SYL and CCh provided additional primary and secondary data, and commented on various drafts of the paper. GC and HK critically reviewed the manuscript.


The Fourth Assessment Report (AR-4) of the Intergovernmental Panel on Climate Change (IPCC) states that most of the observed increase in the global temperatures since the mid-20th century was very likely due to the increase in concentration of anthropogenic greenhouse gases (GHGs).. and natural resources  for their subsistence and livelihoods. The contribution of India to the cumulative global COThe latest high-resolution climate change scenarios and projections for India, based on Regional Climate Modeling (RCM) system, known as PRECIS developed by Hadley Center and applied for India using IPCC scenarios A2 and B2 shows an annual mean surface temperature rise by the end of century, ranging from 3 to 5°C under A2 scenario and 2.5 to 4°C under B2 scenario, with warming more pronounced in the northern parts of India., clean development mechanism (CDM), and emission trading (ET). Only CDMIndia has completed four nationally coordinated assessments of climate change projections, impacts, and mitigation. Recently, National Action Plan on Climate Change (NAPCC) has also been released with many good initiatives including a proposal for a National Solar Mission. Key Ministries of Government of India, such as Environment and Forest, Water Resources and Health and that of states, should devise urgent policy initiatives and implementation of NAPCC to mitigate the effect of climate change in country. Green technology, projects, and industries must be encouraged and promoted by instituting awards and others must be taxed. It must be supplemented by individual efforts leading to collective efforts for saving energy. There is need to promote the use of non-motorized transport systems , mass rapid transport system, and fewer private vehicles by taxing more than one or two vehicles per family for reducing GHG emissions, improving air quality, and making more people physically active. Such an approach would also help in reducing the emerging epidemic of non-communicable diseases including road-traffic accidents. Public health associations in India such as Indian Public Health Association and Indian Association of Preventive and Social Medicine must take lead by organizing thematic seminar and special sessions in conferences. Thus, India has a significant stake in scientific advancement as well as an international understanding to promote mitigation and adaptation. This requires improved scientific understanding, capacity building, networking, and broad consultation processes. Let us join hands to save planet earth and secure our survival and of future generations.


Anopheles and Culex larvae is common, which is likely to impact the transmission of lymphatic filariasis and malaria by the resulting adult mosquito populations. However, the importance of drains as larval habitats remains unknown.Dar es Salaam has an extensive drain network, mostly with inadequate water flow, blocked by waste, causing flooding after rainfall. The presence of Data on mosquito larval habitats routinely collected by the Urban Malaria Control Program (UMCP) and a special drain survey conducted in 2006 were used to obtain a typology of habitats. Focusing on drains, logistic regression was used to evaluate potential factors impacting the presence of mosquito larvae. Spatial variation in the proportion of habitats that contained larvae was assessed through the local Moran's I indicator of spatial association.Anopheles larvae presence and the second highest of Culex larvae presence. However, the majority of aquatic habitats were drains (42%), and therefore, 43%  of all culicine and 33% (320/976) of all anopheline positive habitats were drains. Compared with drains where water was flowing at normal velocity, the odds of finding Anopheles and Culex larvae were 8.8 and 6.3 (p<0.001) times larger, respectively, in drains with stagnant water. There was a positive association between vegetation and the presence of mosquito larvae (p<0.001). The proportion of habitats with mosquito larvae was spatially correlated.More than 70% of larval habitats in Dar es Salaam were human-made. Aquatic habitats associated with agriculture had the highest proportion of Restoring and maintaining drains in Dar es Salaam has the potential to eliminate more than 40% of all potential mosquito larval habitats that are currently treated with larvicides by the UMCP. The importance of human-made larval habitats for both lymphatic filariasis and malaria vectors underscores the need for a synergy between on-going control efforts of those diseases. Lymphatic filariasis and malaria are endemic in Dar es Salaam and partially share common vectors, but no synergy exists between their control programs. The vast majority of mosquito breeding habitats in the city is human-made, and therefore could be mitigated through the implementation of a community-based vector control program accompanied by an educational campaign to sensitize the population and promote behavior change. Of particular importance are clogged drains and ditches with stagnant water, which are the most common larval habitat in the city. This problem could be addressed by an environmental management intervention in order to restore and maintain the functionality of drains. Such intervention can potentially contribute to reduce vector-borne disease transmission, but also promote an overall healthier environment, particularly improving the situation of those living under slum conditions, as proposed by the Millennium Development Goal 7. A synergy between efforts to control lymphatic filariasis and malaria, identifying common strategies, combining monitoring activities, optimizing the use of limited financial resources, and carefully evaluating the cost-effectiveness of the joint venture would not only contribute to current goals of lymphatic filariasis and malaria elimination, but also provide important lessons for future integrated control efforts. According to the 2004 update of the Global Burden of Diseases (GBD) Both malaria and LF are mosquito-borne diseases. Afro-tropical LF and malaria vectors show a high ecological plasticity utilizing a broad range of aquatic larval habitats Fast urban growth often challenges government's ability to provide resources and to properly invest in urban planning. As a result, unplanned and unserviced settlements abound, characterized by lack of sanitation and drinking water, precarious housing, overcrowding, unpaved roads, and inefficient or inexistent solid waste collection Historically, environmental manipulation and modification of potential mosquito larval habitats by engineering works  Currently, Dar es Salaam still faces similar problems, augmented by the pace and pattern of urban growth, particularly since the early 1980s Anopheles larval development Anopheles and Culex mosquitoes throughout the year Anopheles were identified as malaria vectors, namely An. gambiae s.l., An. funestus and An. coustaniCulex quinquefasciatus, An. gambiae s.l., and An. funestusA small-scale urban malaria control effort attempted in Dar es Salaam during 1986–1993 identified drains as important sources of Anopheles and Culex larvae and discuss opportunities for a coordinated effort of vector control that could foster a synergy between current programs to control LF and malaria in Dar es Salaam.Here, we analyze the typology of mosquito breeding habitats in Dar es Salaam, to assess the importance of drains as a source of larval habitats for vectors of LF and malaria. We examine the most common characteristics of drains that were associated with the presence of de facto capital of the United Republic of Tanzania, located along the shores of the Indian Ocean. Administratively, the city comprises three municipalities – Ilala, Kinondoni and Temeke – and is divided into 73 wards, 51 of those considered to be urban, according to the National Bureau of Statistics. Wards are further divided into smaller neighborhood units called mitaa  mtaa is subdivided into ten-cell units (TCU), or clusters of approximately 10–20 houses, although some TCUs contain a much larger number of houses.Dar es Salaam is the largest city and 2  During May 2006 and March 2007 we conducted a survey in the study area to assess the physical characteristics of the drains  and their current conditions  was assessed through the use of the local Moran's I indicator of spatial association Spatial variation in the proportion of aquatic habitats that contained Anopheles larvae in surveyed drain segments. In the second model, the outcome variable indicates the presence of Culex larvae in surveyed drain segments. In both models we included independent variables that showed a significant effect in univariate analysis. These independent variables were all binary and included: (i) presence of a human-made connection to the drain segment – these connections are often illegal and tend to leave exposed pipes inside the drain blocking the water flow; (ii) lined drain segment – lined drains are less likely to present erosion; (iii) located in residential areas; (iv) presence of waste materials; (v) drain segment contained vegetation; (vi) efforts to maintain the drain in good conditions were conducted regularly; (vii) drain segment often floods after the rains; (viii) presence of stagnant water; (ix) water flowing at low velocity; (x) drain segment easily accessible by trucks; (xi) drain segment constructed before 1992; and (xii) drain segment surveyed during the dry season. Finally, the first model included a binary variable indicating the presence of Culex larvae, and the second model included a binary variable indicating the presence of Anopheles larvae.We analyzed the UMCP larval habitat survey data to obtain a typology of aquatic habitats in the study area and to assess their seasonal variation due to rainfall. Focusing on drains, we used the drain assessment survey to evaluate potential factors impacting the presence of larvae in these habitats. We fitted two separate logistic regression models. In the first model, the outcome variable indicates the presence of Model goodness-of-fit was assessed through the calculation of three diagnostics measures: (i) the area under the receiver operating characteristic (ROC) curve; (ii) the detection of extreme observations ; and (iii) the assessment of heteroscedasticity in the residuals (robust standard errors) Anopheles larvae, 24% contained Culex larvae, and in 5% the concurrent presence of both anopheline and culicine larvae was observed. A detailed analysis by habitat type (Anopheles (33%), followed by ponds (25%) and habitats associated with rivers and streams (21%). In contrast, anophelines were, on average, found in only 6% of the drains. Overall, a higher proportion of habitats were colonized by culicine larvae than anopheline larvae. Culex larvae were, on average, present in 51% of the ponds, 48% of habitats associated with agriculture, 43% of habitats associated with rivers and streams, but in only 25% of the drains. Notably, the vast majority of the weekly surveyed aquatic habitats were drains (42%) and only 3% of the habitats were associated with agriculture. Therefore, 33% (320/976) of all anopheline positive habitats and 43%  of all culicine positive habitats found per weekly survey were drains and ditches, three and eight times more than anopheline and culicine positive agricultural sites, respectively  contained water in one or more of the surveyed weeks . On avertat type  indicateectively . Construectively , and com puddles . Heavy rAnopheles larvae were observed in TCUs located in the northern part of the study area (Mikocheni ward), in the western part (Kipawa ward), and in the eastern-most area  times larger, and those with stagnant water were 8.3 (95% CI 5.8–11.7) times larger. Drain segments with vegetation were 3.4 (95% CI 2.5–4.6) times more likely to contain Anopheles larvae. Finally, Anopheles larvae were more likely to occur in drain segments that also contained Culex larvae . An important difference between characteristics that impacted the presence of Anopheles and Culex larvae was related to the occurrence of accumulated waste materials in drains .Results from the drain assessment survey indicated almost complete absence of maintenance activities, common presence of waste materials inside the drains, inadequate water flow, occurrence of flooding during the rainy season, and presence of x larvae . Importacurrence , Model 1n drains , Model 2Model diagnostics indicated a good fit, as represented by a ROC value of approximately 0.93 . In addiAnopheles and Culex larvae in the city of Dar es Salaam. Nevertheless, drains seem not to be the most preferred habitat by these species, since a small proportion contained larvae. In contrast, the much fewer aquatic sites associated with urban agriculture are more likely to be found with larvae of both mosquito species. In addition, the vast majority of larval habitats in Dar es Salaam are human-made. Drains, borrow pits, and house foundations under construction comprised 55% of all habitats that contained Anopheles larvae and 69% of all open habitats that contained Culex larvae. These habitats are direct consequences of human actions, as opposed to puddles, swampy grounds, marshes and seepages which are mainly a result of the local environmental conditions.Dar es Salaam is served by an extensive network of drains , designed to lower the water level in the city and to prevent the accumulation of stagnant water suitable for vector proliferation Anopheles and Culex larval habitats. These results highlight that, under ideal conditions, drains should serve as a tool for source reduction of all mosquito types, including the vectors of numerous neglected tropical diseases, and therefore it is not the habitat per se that is conducive to mosquito breeding, but human activity and lack of maintenance that can produce ideal conditions for larval development.In well maintained drain segments, where water can flow undisrupted and vegetation is absent, the likelihood of finding anopheline larvae was reduced by over 90%. Culicines showed a wider distribution in drain segments with various characteristics, particularly waste accumulation. Interestingly, there was a very strong positive association between the presence of anophelines and the presence of culicines. Similar findings have been reported from habitats in rural areas in East and West Africa 2), located in lowland areas, close to rivers or ponds but far from drains, and with loamy or clayey soils Anopheles and Culex larvae were observed in three out of the 15 surveyed city wards. This spatial variability is likely to be a result of idiosyncratic interactions between the local ecology and human behaviors that have the potential to minimize or augment the negative impacts of human-made transformations on vector development. Spatial exploratory approaches similar to those utilized in this analysis, as well as more sophisticated spatial modeling, should be applied more frequently in entomological studies to better capture the occurrence of local patterns in the distribution of positive habitats, and to shed light on the likely factors that determine such patterns.The typology of breeding habitats did vary spatially. Some habitat types, such as ponds, agricultural sites, and drains are located in specific areas, and are not randomly distributed across the city. In the case of agriculture, previous studies showed that the presence of larvae was more likely in larger fields  or by their size  even though the spatial variation in habitat abundance is likely to be partially influenced by these variables. However, the clustering pattern demonstrated in our analysis is particularly important as a first step toward better understanding of the local variations of human exposure to LF and malaria, and could be utilized for more targeted interventions The majority of open vector breeding sites could be readily avoided by keeping the drain network in Dar es Salaam in good condition, through the implementation of routine EM activities, embedded in an integrated vector management (IVM) approach These activities would greatly benefit from involvement of community members Such multidisciplinary and community-based approaches applied to contemporary EM interventions would represent an important distinction from historical EM efforts, which were mainly vertical (and often authoritarian) programs A community-based pilot EM intervention conducted in Dar es Salaam in 2008 provided initial evidence of the potential to increase awareness and local engagement after sensitization campaigns Currently, there is a lack of synergy between the National Malaria Control Program (NMCP)/UMCP and the National Lymphatic Filariasis Elimination Program (NLFEP), although opportunities for combined action do exist. Strategies of the NMCP in Dar es Salaam include the introduction of malaria rapid diagnostic tests in health facilities, the distribution of long-lasting insecticidal nets, the improvement of intermittent preventive treatment of malaria in pregnancy, and wide access to artemisinin-based combination therapy Considering the importance of drains as larval habitats for both LF and malaria vectors, and the often constrained availability of financial resources in countries endemic to both diseases http://www.undp.org/mdg/goal7.shtml). Further studies are needed to provide definite evidence of such potential successful outcomes.In 1997, the World Health Assembly called for LF elimination after the International Task Force for Disease Eradication identified the disease as potentially eradicable