Improved usual gait speed predicts substantial reduction in mortality. A better understanding of the modifiable factors that are independently associated with improved gait speed would ensure that intervention strategies are developed based on a valid theoretical framework. Thus, we examined the independent association of change in executive functions and change in falls-related self-efficacy with improved gait speed among community-dwelling senior women.A secondary analysis of the 135 senior women aged 65 to 75 years old who completed a 12-month randomized controlled trial of resistance training. Usual gait speed was assessed using a 4-meter walk. Three executive processes were assessed by standard neuropsychological tests: 1) set shifting; 2) working memory; and 3) selective attention and response inhibition. A linear regression model was constructed to determine the independent association of change in executive functions and falls-related self-efficacy with change in gait speed.Improved selective attention and conflict resolution, and falls-related self-efficacy, were independently associated with improved gait speed after accounting for age, global cognition, baseline gait speed, and change in quadriceps strength. The total variance explained was 24%.Interventions that target executive functions and falls-related self-efficacy, in addition to physical functions, to improve gait speed may be more efficacious than those that do not.ClinicalTrials.gov Identifier: NCT00426881 Gait speed is a global indicator of functional mobility and status. Reduced speed is associated with risk of major health-related outcomes , includiAlthough there has been a call for research to determine whether interventions to improve gait speed affect survival , a betteCurrent evidence suggests that executive functions should be considered in interventions to improve gait speed in older adults. The Health, Aging and Body Composition Study  highlighExecutive functions are higher order cognitive processes that control, integrate, organize, and maintain other cognitive abilities . These cEvidence from neuro-imaging studies provides insight to possible mechanisms underlying the association between executive functions and gait speed. Specifically, cerebral white matter lesions -- which are associated with reduced executive functioning  - are alFalls-related self-efficacy is associated with gait performance ,22 and hchange in executive functions and falls-related self-efficacy are independently associated with change in gait speed in older adults after accounting for relevant factors such as age and global cognitive function. Yet, such knowledge would facilitate the development and refinement of targeted interventions to improve gait speed in older adults. For example, if improved executive functioning were independently associated with improved gait speed, it would be logical to incorporate cognitive training that focused specifically on executive functioning rather than other domains of cognition such as memory. Thus, we examined the independent association of change in three key executive processes and change in falls-related self-efficacy with improved gait speed among community-dwelling senior women after accounting for age, global cognition, baseline gait speed, and change in quadriceps strength.To our knowledge, it is currently unknown whether The sample for this analysis consisted of 135 women who consented and completed a 12-month randomized controlled trial of exercise that aimed to examine the effect of once-weekly or twice-weekly resistance training compared with a twice-weekly balance and tone exercise intervention on cognitive performance of executive functions. The design and the primary results of the study have been reported elsewhere .We recruited women who: 1) were aged 65 to 75 years; 2) were living independently in their own home; 3) scored > 24 on the Mini-Mental State Examination (MMSE); and 4) had a visual acuity of at least 20/40, with or without corrective lenses. Recruitment strategies have been described previously . Ethical® Pressurized Air system and free weights were used to provide the training stimulus. Other key strength exercises included mini-squats, mini-lunges, and lunge walks.All classes were 60 minutes in duration. The protocol for this program was progressive and high-intensity in nature. Both a KeiserThis program consisted of stretching exercises, range of motion exercises, kegals, balance exercises, and relaxation techniques. This group served to control for confounding variables such as physical training received by traveling to the training centres, social interaction, and lifestyle changes secondary to study participation.Global cognition was assessed using the MMSE . We usedParticipants were asked to walk at their usual pace along a 4-meter path. Gait speed (m/s) was calculated from the mean of two trials. The test-retest reliability of gait speed in our laboratory is 0.95 (ICC) .A simple strain gauge was used to assess dominant quadriceps (isometric) strength to the nearest 0.5 kilogram . ParticiFor this study, we referred to the work by Miyake and colleagues  who idenWe used the Trail Making Tests (Part A & B) to assess set shifting . Part A We used the verbal digits forward and verbal digits backward tests to index the central executive component of working memory . Both teWe used Graf, Uttl and Tuokko's  version The 16-item Activities-specific Balance Confidence (ABC) Scale  assessedDescriptive data are reported for variables of interest. Data were analyzed using SPSS Windows Version 17.0 . The associations between the variables were determined using the Pearson product moment coefficient of correlation.A multiple linear regression model was constructed to determine the independent association of change in executive processes and change in falls-related self-efficacy with change in gait speed over a 12-month period. Change in gait speed, falls-related self-efficacy, and dominant quadriceps strength was calculated by subtracting the baseline value from the trial completion value. Change in the three executive functions was calculated by subtracting the trial completion value from the baseline value. Experimental group, age, global cognition, baseline gait speed, and the change in dominant quadriceps strength were statistically controlled by entering these four variables into the regression model first. These independent variables were determined from the results of the Pearson product moment coefficient of correlation analyses and based on biological relevance, such as experimental group, baseline global cognition, and change in dominant quadriceps strength.P < 0.05.The three executive processes were then entered into regression model and only those that significantly improved the model were kept in the model . The ABC Scale score was entered last into each model. Alpha was set at Of the 155 participants who consented and were randomized at baseline, 135 completed the 12-month exercise trial. Table P < 0.001).At the end of the 12-month trial, the 135 women demonstrated a mean increase of 7 kilograms in dominant quadriceps strength and a four- and nine-second reduction in Stroop Test and Trails Making Tests completion time, respectively (Table P < 0.01). Change in falls-related self-efficacy was also significantly and positively associated with improved gait speed (P < 0.001).Table F Change = 11.6, P = 0.001). Adding change in the ABC Scale score to the model resulted in an R-square change of 4.5% and significantly improved the model . The total variance accounted by the final model was 24.0% (Table Experimental group, age, baseline gait speed, baseline global cognition, and change in dominant quadriceps strength accounted for 11.9% of the variance in improved gait speed Table . Adding 0% Table .There is an increasing recognition that gait speed is an important indicator of current and future functional status in older adults. We found that both improved cognitive performance of selective attention and conflict resolution and falls-related self-efficacy were independently associated with improved gait speed among community-dwelling senior women. To our knowledge, this is the first study that has examined the independent association of changes in executive functions and falls-related self-efficacy with improved gait speed after accounting for age global cognition, baseline gait speed, and change in dominant quadriceps strength.Our findings concur with previous studies demonstrating the association between executive functions and gait speed ,35. Of tAs gait speed is an independent predictor of falls  and fracOur finding of an independent association between improved falls-related self-efficacy and improved gait speed extend those of two previous cross-sectional investigations ,22. SpecAccording to the LIFE-P study , the obsOur finding that improved dominant quadriceps strength was not significantly associated with improved gait speed concurs with previous findings . A modelAn important clinical implication of these results is that clinicians may need to consider the specific executive process of selective attention and conflict resolution and falls-related self-efficacy in the rehabilitation of gait among community-dwelling seniors. Our data suggest interventions that target executive functions and falls-related self-efficacy in addition to physical functions to improve gait speed may be more efficacious than those that do not.We note that our study sample consisted exclusively of independent community-dwelling senior women who were without significant physical and cognitive impairments and without a significant history of falls. Thus, the results of our study may not generalize beyond this population of senior women and we may have underestimated the contribution of change in executive functions and falls-related self-efficacy to change in gait speed. Future prospective studies are needed to test whether the present findings also apply to larger, more heterogeneous populations. Finally, because the effect of selective attention and conflict resolution and falls-related self-efficacy observed in this study was small, future studies are also needed to determine the clinical significance, if any, of the additional and unique variance accounted for by each factor.In conclusion, both change in executive functions and falls-related self-efficacy are independently associated with improved gait speed. Clinicians may need to consider these factors when developing rehabilitation strategies to improve gait speed.The authors declare that they have no competing interests.TLA conceived and designed the study, acquired data, and analyzed and interpreted the data. TLA, JCD, and KMK drafted and revised the manuscript. LSN, LK, and CLH acquired data and participated in the statistical analysis. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2318/10/25/prepub


Advances in automated DNA sequencing technology have accelerated the generation of metagenomic DNA sequences, especially environmental ribosomal RNA gene (rDNA) sequences. As the scale of rDNA-based studies of microbial ecology has expanded, need has arisen for software that is capable of managing, annotating, and analyzing the plethora of diverse data accumulated in these projects.XplorSeq is a software package that facilitates the compilation, management and phylogenetic analysis of DNA sequences. XplorSeq was developed for, but is not limited to, high-throughput analysis of environmental rRNA gene sequences. XplorSeq integrates and extends several commonly used UNIX-based analysis tools by use of a Macintosh OS-X-based graphical user interface (GUI). Through this GUI, users may perform basic sequence import and assembly steps , perform BLAST  searc.XplorSeq should benefit researchers who are engaged in analyses of environmental sequence data, especially those with little experience using bioinformatics software. Although XplorSeq was developed for management of rDNA sequence data, it can be applied to most any sequencing project. The application is available free of charge for non-commercial use at  The recent explosions in culture-independent studies of environmental DNA sequences ("metagenomics") and automated DNA sequencing capabilities have prompted the creation of numerous software applications designed to aid the analysis of an avalanche of sequence data. However, many of the commonly used, freely available applications require some facility with the UNIX/Linux operating system and/or specialized scripting languages to either manipulate files in batch or pipe data between applications. As automated DNA sequencing and sequence analysis has become commonplace in laboratories that do not specialize in bioinformatics, need has arisen for the development of powerful, yet simple-to-use, software.XplorSeq, written for the Macintosh OS X operating system, provides a graphical user interface (GUI) that integrates the use of several popular UNIX-based DNA sequence analysis applications. A number of additional features have been incorporated in order to track, annotate, and analyze sequence information in a manner conducive to high-throughput metagenomics. Implementation of a common GUI Fig.  eliminat.XplorSeq was developed for rapid compilation and analysis of rDNA clone libraries, but should be applicable to any sequencing project . Although several commercial and non-commercial software packages implement some of the same basic functionalities as XplorSeq, the development of XplorSeq was motivated by the absence of GUI-based software designed specifically for high-throughput, batch analysis of rDNA sequences, such as arise from culture-independent metagenomic studies. Specifically, the extant software could not accommodate the phylogenetic orientation of analyses and sequence annotations that are most useful for metagenomics. In contrast, XplorSeq implements several domain-specific software tools  that are not available in general-purpose DNA analysis packages. Many published studies, from a variety of laboratories engaged in metagenomics, have used XplorSeq, and thereby established its stability, ease-of-use, and capabilities -29. The XplorSeq is written in Objective-C using the Cocoa application framework (Apple Inc.). Releases are compiled for the OS X operating system (current versions require OS 10.4.x or 10.5.x) as universal binaries, which run natively on Macintosh computers with Intel or PowerPC microprocessors. Similar to Cocoa, the architecture of XplorSeq is based on the Model-View-Controller (MVC) design pattern. XplorSeq is multi-threaded and can adjust its operation to accommodate multiple shared-memory microprocessors.The rationale for implementing XplorSeq as a standalone Macintosh application involved 1) desire for a highly responsive, feature-rich graphical output, thus precluding web-based applications; 2) recognition that the BSD-Unix operating system underpinning OS X would allow leveraging of existing open-source software; 3) observation that many computer novices (an intended audience for this software) were more comfortable with OS X than other operating systems; and 4) the maturity, stability, and support inherent in the Cocoa application framework.).Third-party software packages and plugin executables (sortx and biodiv) were written in C and C++. When possible, compiled executables are incorporated directly into the XplorSeq application bundle  so that users can install and operate XplorSeq without the need for local compilation or extensive configuration. Full implementation of XplorSeq requires separate installation of phred and phrap  hard-coded data objects (i.e. "BlastInfo" and "Oligo" objects) and 2) a customizable "Metadata" object. Metadata objects are implemented as key-value dictionaries  that can be linked to Project, Clone, and Sequence data objects. Users specify keys and values for particular objects through import of tab-delimited spreadsheets in which the first column designates the names of objects (i.e. sequence/clone names) and subsequent columns specify values (keyed by column headings) to attach to the objects. Alternatively, object inspector windows provide GUI-based features in which to display and edit meta-data ; calculate distance matrices (phylip dnadist [Table  dnadist ); constr dnadist ; phylip  dnadist ; RAxML,  dnadist ); and ca or created by use of the formatdb executable (through XplorSeq or the command line). XplorSeq dispatches sequences to the appropriate client software  and tXplorSeq provides graphical interfaces for several commonly used command-line phylogenetic tools are shown in Fig. and sequence length . Finally, the user can select a range of pairwise sequence distance thresholds by which to assemble OTUs in order to create multiple data sets at different phylogenetic depths.OTU clustering is implemented through the program sortx, which was written in tandem with XplorSeq Fig. . Sortx uobs, Schao1, CACE, Good's coverage, Shannon diversity; [Estimates of biodiversity indices  can be reported through either of two modes. First, the export function OTU Diversity... reports basic calculations of commonly used indices  for a sversity; . Biodiv versity; .XplorSeq provides several features by which to export sequences and data for further analyses. Data reports are generated in tab-delimited format for import into spreadsheets. A particularly useful export feature tabulates sequence abundances and/or prevalences on the basis of user-defined terms that define how data are categorized and enumerated. For instance, as shown in Fig. Preparation of sequence data for submission to public databases can be tedious and error-prone, despite the availability of tools such as NCBI's sequin software tool. XplorSeq can facilitate the submission process by automating the organization of sequences and their annotations into a form suitable for input into sequin Fig. . The useFor detailed phylogenetic studies, sequences assembled by XplorSeq can be exported in formats suitable for input into other software packages, such as ARB , DOTUR , NAST 44, SILVA rd party software  are invoked in separate threads, so that multiple tasks can be run in parallel. During these steps, feedback to the GUI is limited in order to minimize the overhead of performing analyses through XplorSeq. Typical latencies associated with XplorSeq contribute no more than an additional ~10% to the total elapsed time required for an analysis, relative to the same task performed through command line execution of the underlying 3re Table . This ovThe XplorSeq file format, implemented using the Cocoa software framework, does not significantly add to the size of data files. For example, an XplorSeq file containing an alignment of 250,000 rRNA sequences (1585 nucleotides per sequence) requires 396 megabytes (MB) of storage, compared to 381 MB for a fasta formated file storing the same alignment. The addition of metadata, such as blast results, increases file size in roughly linear proportion to the size of parsed input text.rd party software tools as by XplorSeq. However, the XplorSeq environment is scalable from laptops to more advanced workstations  so its capabilities can be expanded as need arises and hardware evolves.Although the use of a flat file format by XplorSeq greatly simplifies data storage and transfer, it does require that all data be read into memory before being manipulated. For relatively large files, bottlenecks are apparent primarily in tasks requiring import and export of data. Thus, sequence analysis projects are likely to be limited as much by computer hardware  and the performance of underlying 36 25-nucleotide sequences can be manipulated with similar lag times. More fully annotated files containing ~50,000 16S rRNA sequences, each of length ~1000 nucleotides, have been used routinely on a laptop with little performance degradation [As a rough guide to system requirements, Table radation ,23,27. HAlthough XplorSeq was developed to expedite the phylogenetic analysis of ribosomal RNA (rRNA) gene libraries, it should prove useful in any sequencing project, particularly ones facilitated by batch analysis of multiple clones. Moreover, any UNIX-based DNA sequence analysis tool that can be ported to Mac OSX can be readily incorporated into XplorSeq. Suggestions for the addition of other modules to the XplorSeq package are most welcome.Project name: XplorSeqProject home page: Operating system: Macintosh OS X (currently requires 10.4.x or 10.5.x)Programming language: Cocoa/Objective-C, C, C++Other requirements: phred and phrap are available at License: Daniel N. Frank. Free for non-commercial useAny restrictions to use by non-academics: Contact corresponding author. Users are requested to notify the corresponding author when XplorSeq is cited.Example of sequence enumeration table.Click here for file


Silvetia compressa. Previously, this motor was shown to be needed to assemble a bipolar spindle, similar to animals. This report builds on those findings by investigating the localization of the motor and probing its function in spindle maintenance.Kinesin-5 (Eg-5) motor proteins are essential for maintenance of spindle bipolarity in animals. The roles of Kinesin-5 proteins in other systems, such as Arabidopsis, Dictyostelium, and sea urchin are more varied. We are studying Kinesin-5-like proteins during early development in the brown alga Anti-Eg5 antibodies were used to investigate localization of Kinesin-5-like proteins in brown algal zygotes. In interphase zygotes, localization was predominantly within the nucleus. As zygotes entered mitosis, these motor proteins strongly associated with spindle poles and, to a lesser degree, with the polar microtubule arrays and the spindle midzone. In order to address whether Kinesin-5-like proteins are required to maintain spindle bipolarity, we applied monastrol to synchronized zygotes containing bipolar spindles. Monastrol is a cell-permeable chemical inhibitor of the Kinesin-5 class of molecular motors. We found that inhibition of motor function in pre-formed spindles induced the formation of multipolar spindles and short bipolar spindles.S. compressa serve novel roles in spindle formation and maintenance not observed in animals.Based upon these localization and inhibitor studies, we conclude that Kinesin-5-like motors in brown algae are more similar to the motors of animals than those of plants or protists. However, Kinesin-5-like proteins in  Kinesins are a diverse group of molecular motors present in protozoans, fungi, plants, and metazoans . They shMonastrol is a cell permeant inhibitor of Kinesin-5 motors and is thought to function by binding the motor domains, thereby blocking normal movement . SpecifiSilvetia compressa zygotes prior to mitosis induced the formation of mostly multipolar spindles; monasters were also formed albeit to a lesser degree [It has previously been shown that monastrol treatment leads to malformed spindles in brown algae. Monastrol treatment of r degree . MonastrS. compressa zygotes. We show that Kinesin-5-like motors are localized within the nucleus during interphase, and strongly localize to the spindle poles from the onset of mitosis until nuclear envelope reformation. These motors are also associated with polar microtubules and the spindle midzone but to a much lesser degree. Additionally, we show that inhibition of Kinesin-5-like motors during mitosis leads to the formation of multipolar and short bipolar spindles, indicating that motor function is needed to maintain spindle integrity. Many of our findings are in concert with those found in animal cells. In both brown algal zygotes and animal cells, Kinesin-5 proteins are localized within the nuclear envelope until mitosis where they strongly decorate the spindle poles, and monastrol treatment induces formation of aberrant spindles.Here we examine Kinesin-5-like localization during early embryonic development and utilize monastrol to assess motor function in maintaining spindle structure and function in 2, pH adjusted to 7.5 with KOH) containing 3% paraformaldehyde and 0.5% glutaraldehyde. Fixed zygotes were processed as previously described [Zygotes were cultured as previously described . Monastrescribed . KinesinS. compressa. During interphase of the first cell cycle, anti-Kinesin-5 signal was observed exclusively within the nucleus : after fertilization; (ASW): artificial sea water.The authors declare that they have no competing interests.NTP and ACM carried out the experiments, NTP and DLK wrote the manuscript, and DLK raised the funds. All authors have read and approved the final manuscript.


Oestrogen exerts a robust yet imperfectly understood effect on sexual development in vertebrate embryos. New work by Pask and colleagues in BMC Biology indicates that it may interfere with male development by preventing nuclear localization of SOX9, a master regulator of the testis differentiation pathway.http://www.biomedcentral.com/1741-7007/8/113See research article  BMC Biology, Pask et al. [Vertebrates exposed to exogenous oestrogens during embryonic development often exhibit extensive sexual abnormalities as adults, ranging from genital and reproductive tract malformations to infertility or full male-to-female sex reversal. Although these effects have been studied at the phenomenological level for decades, no consensus has emerged regarding the molecular details of how this hormone influences primary sex determination, the process in which the initially bipotential gonads commit to testis or ovary differentiation. In the present issue of k et al.  show thaNon-mammalian vertebrate embryos are particularly sensitive to oestrogen-induced male-to-female sex reversal and reproductive dysgenesis. Suppression of oestrogen synthesis in these animals has the opposite effect, causing predetermined females to develop as males. These findings collectively indicate that oestrogen is necessary and sufficient for female development in most non-mammalian vertebrates. While this may be an advantageous system under some circumstances, the accumulation of oestrogenic chemicals as a result of agricultural runoff and pollution from modern pharmaceutical and industrial practices can have grave consequences on sexual development in egg-laying species, especially those that lay their eggs near water.Sry, that directs the bipotential gonads to develop as testes  system of sex determination based on the presence or absence of the Y chromosome. XY embryos express a gene on the Y chromosome, iewed in ). XX embon (e.g. ), althouon (e.g. . Convincon (e.g. ). In theHowever, the case is somewhat different in marsupials. Marsupial development is divided into a short period of placental gestation followed by a long period of postnatal development during lactation. Sex determination in this group of mammals also depends on the XY-SRY system, but the process occurs around the time of parturition, not mid-gestation as in eutherians (reviewed in ). Marsupet al. [SOX9, was not down-regulated in the oestrogen-treated samples. SOX9 (SRY-box containing gene 9) is a highly conserved HMG-box transcription factor that in mice is directly activated by SRY and can induce full female-to-male sex reversal when misexpressed in XX embryos . Its fuiewed in .et al. examined SOX9 protein localisation in the oestrogen-treated marsupial gonads, they saw that the protein was indeed still present, but diffuse and cytoplasmic, in sharp contrast to its normal dense nuclear pattern . OestroSox9 transcription in the adult mouse ovary [In the red-eared slider turtle, SOX9 is initially expressed at high levels during the bipotential period in both presumptive sexes, then declines precipitously at the onset of ovarian differentiation [se ovary . These fAlthough sex determination in marsupials retains the ancestral sensitivity to oestrogen, it is not clear that oestrogen is involved in the endogenous process of making an ovary in these animals. The β-estradiol content of developing tammar gonads is minimal , and no


To examine the effects of short duration stress management training (SMT) on self-perceived depression, anxiety and stress in male automotive assembly workers, 118 male automotive workers from Pekan, Pahang  and Kota Bharu, Kelantan  were assigned to experimental and control group, respectively. A SMT program consisting of aerobic exercise, stress management manual, video session, lecture, question and answer session, and pamphlet and poster session were conducted in the experimental group. A validated short-form Malay version of the Depression Anxiety Stress Scales (DASS-21) were self-administered before and after the intervention program in the experimental and control group and their time and group interaction effects were examined using the repeated measure ANOVA test. Results indicated that the mean (SD) scores for DASS-Depression (p = 0.036) and DASS-Anxiety (p = 0.011) were significantly decreased, respectively, after the intervention program in the experimental group as compared to the control group (significant time-group interaction effects). No similar effect was observed for the mean (SD) scores for DASS-Stress (p = 0.104). However, the mean (SD) scores for subscales of DASS-Depression , DASS-Anxiety , and DASS-Stress  showed significant time and group interaction effects. These findings suggest that short duration SMT is effective in reducing some aspects of self-perceived depression, anxiety and stress in male automotive workers. Job stress is a major occupational health problem in many sectors of industries and automotive assembly industry workers are one of several occupational groups who report disproportionately high levels of job stress ,2. StudiSeveral stress intervention programs have been developed over the years to counter health problems in the workplace. According to van der Klink et al. , interveThere are various techniques in the individual-focused approach that include exercise, stress management, relaxation training, physical fitness, cognitive-behavioural training, meditation, biofeedback, hypnosis, yoga, and interpersonal skills. A recent meta-analysis reported that job stress management intervention studies that focus on the individuals are effective in reducing workers' stress-related complaints . Among tth Malaysia Plan.A quasi experimental study was conducted among male automotive workers in Pahang and Kelantan from July to September, 2006. This study is part of a national research project entitled "Occupational Stress Intervention Study in Petrochemical and Automotive Assembly Plants: Developing and Evaluating a Stress Management Program at Workplaces" (OSIS) under the research program "Quality of Work Life – National Occupational Risk Management Study and National Injury Prevention" (NORMS) funded by the Ministry of Science, Technology and Innovation (MOSTI), Malaysia under its 8Sample size was estimated using the PS Software for two sample means, with power of 80% and alpha of 5%. Sample size calculation was based on the standard deviation (SD = 10.12) for self-perceived stress in general practice patients with mental health problems in Australia  and the The reference population consists of automotive workers in Malaysia. The source population includes workers in an automotive plant in Pekan, Pahang and workers in an automotive plant in Kota Bharu, Kelantan. The sampling frame is the list of automotive workers available for screening using the DASS questionnaire in Pekan, Pahang (88 workers) and Kota Bharu, Kelantan (75 workers). Inclusion criteria include male automotive workers and at least one year of working experience. Exclusion criterion is a history of medically confirmed mental illnesses. Based on the inclusion and exclusion criteria, 60 automotive workers were eligible in Pekan, Pahang  and 58 automotive workers in Kota Bharu, Kelantan (assigned to the control group).Our intervention program consists of a short duration SMT module that includes the following sessions: aerobic exercise, pamphlet and poster session on stress reduction through adopting a healthy lifestyle, review session of a simplified and well illustrated manual on the prevention and control of stress in the workplace, interactive lecture session on stress management in the workplace, video session on stress management in the workplace, and question and answer session. The intervention program was held in a public facility adjacent to the automotive plant in Pekan, Pahang.Aerobic Exercise. The aerobic exercise session was conducted by qualified aerobic instructors and consists of a preparatory 15 minutes warming up stretching exercises . This waPamphlet and Poster Session. This is an interactive pamphlet and poster session where participants were guided by the researchers to the pamphlets and posters that depict the nature, causes, and signs and symptoms of stress, stress reduction through adopting a healthy life style, and methods of managing stress in the workplace. A short question and answer session was conducted to improve the participants' understanding of the materials presented.Manual Review Session. A simplified and well illustrated manual on the prevention and control of stress in the workplace was distributed to all participants and they were asked to review it over the next 30 minutes followed by a question and answer session.Lecture on Stress Management. This is an interactive lecture session on stress management in the workplace and designed to reinforce the participants' understanding of occupational stress and a suggested plan towards stress reduction and control. The key component of the lecture is an elaboration of occupational stress, risk factors involved, associated health impacts and ways to prevent and control stress in the workplace. References were made to the Job Demand Control Model of stress and how work conditions of high job demand, low job control and poor social support can result in severe stress and serious health consequences. Participants were carefully guided as to the various ways of dealing with stress at work and the importance of good social support in protecting workers from severe stress.Video Session on Stress Management in the Workplace. The video session is entitled "Stress Management in the Malaysian Automotive Industry" and was produced by the researchers as part of the research program, with permission and full cooperation from the automotive assembly plant. The video previewed all processes of production in the automotive industry. The aim of this session is to provide an overview of the working conditions in the automotive assembly plant and how the working conditions can contribute toward occupational stress. The video also showed several simple methods of dealing with stress in the workplace.Question and Answer Session. At the end of the video session, the researchers conducted a question and answer session with the participants. This session was aimed at exploring several practical problems related to occupational stress encountered by the participants and discussing several options and suggestions in managing these problems. Participants were asked to identify several stressful situations that they had personally experienced at work and how they cope with these stresses. Suggestions were made to strengthen their coping skills including the need to adopt the healthy lifestyle approach – regular exercise, avoidance of tobacco, drugs, and alcohol, adequate sleep and rest, and proper nutrition and dieting – that has been shown to be beneficial in stress reduction.DASS-21 is the short-form of Lovibond and Lovibond's 42-item self-report which measures the negative emotional states of depression, anxiety and stress . The queTo examine the effect of short duration SMT intervention program on self-perceived depression, anxiety and stress, the DASS-21 questionnaires were self-administered to both study groups before and after the intervention program.Data entry and analysis was done using SPSS version 12.0.1 . Mean and standard deviations (SD) were calculated for continuous variables, and frequencies and percentages for categorical variables. Effectiveness of the short duration SMT program was examined using the repeated measure ANOVA analysis where the time-group interaction effect (within subject-between subject) was modelled. Assumptions of the repeated measure ANOVA include the following: histogram of residuals for normality assumption and Levene's test for equal variance assumption; sphericity assumption is not necessary because the model includes only 2 repeated measures; model fitness will be checked by residual plots and lack-of-fit test; level of significance is set at α = 0.05.A total of 118 workers  participated in this study. Socio-demographic characteristics such as age, marital status, years of education, duration of work and salary were compared between the two groups (Table p = 0.036) and DASS-Anxiety (p = 0.011) in the experimental group as compared to those in the control group (significant time-group interaction effects). However, there is no significant difference in the mean (SD) scores (pre- and post-intervention) for DASS-Stress (p = 0.104) in the experimental group as compared to those in the control group (not significant time-group interaction effect).Table p = 0.010) but not for evaluation of life (p = 0.200), hopelessness (p = 0.370), self-deprecation (p = 0.076), anhedonia (p = 0.352) and inertia (p = 0.350). Meanwhile, for DASS-Anxiety Subscales, there are significant time-group interaction effects in the mean (SD) scores for anxiety (p = 0.007) and situational anxiety (p = 0.048), but not for skeletal muscle effect (p = 0.495) and autonomic arousal (0.054). Whereas for DASS-Stress Subscales, there are significant time-group interaction effects in the mean (SD) scores for nervous arousal (p = 0.018) and easily upset (p = 0.047) but not for difficulty relaxing (p = 0.750) and irritable (p = 0.722).Table We examined the effects of short duration SMT on self-perceived depression, anxiety and stress in automotive workers using the validated short-form Malay version of the Depression Anxiety Stress Scales (DASS-21). Results indicated that SMT is effective in reducing some aspects of self-perceived depression, anxiety, and stress in these workers.p = 0.02), depressive symptoms as measured by the Beck Depression Inventory (p = 0.02) and improved markers of cardiovascular risk more than usual medical care alone. Our present study also shows that short duration SMT is effective in reducing some aspects of self-perceived depression, anxiety and stress in automotive workers. These findings are very important because automotive assembly workers tend to be busy, and long duration of training would be less likely to be used. However, it should be noted that our study did not consider compliance or continuation of the present training. Further research must be carried out to explore how the subjects would respond to longer duration training of weeks or even months.Blumenthal et al  have shop = 0.03). However, no significant relaxation training effects were found in the Profile Mood States (POMS) subscales for tension-anxiety (p = 0.870), depression-dejection (p = 0.161), anger-hostility (p = 0.937), vigor-activity (p = 0.657) and fatigue-inertia (p = 0.212), except for confusion (p = 0.036). Since the scope of the present study is only focused on responses using self-perceived DASS21 questionnaire, we suggest that further studies using physiological responses are needed to explore the effectiveness of SMT.To our knowledge, there is only one empirical study that examined the immediate effects of an intervention program . Tanigucet al. [Studies that examine the effects of short duration SMT on stressful workers are seriously lacking. One reason for this lack is that senior managers in the same organization presume that formal exercise training and SMT programs are time-consuming, expensive and workers cannot spare the time to practice any coping skills. Another reason is that provided by Yung et al. , with gip = 0.010) was significantly reduced after intervention. This could be due to the effect of the SMT intervention program, especially the aerobic exercise. However, the mean (SD) scores (pre- and post-intervention) for DASS-Depression subscales such as devaluation of life (p = 0.200), hopelessness (p = 0.370), self deprecation (p = 0.076), anhedonia (p = 0.352) and inertia (p = 0.350) were not significantly different in the experimental group as compared to those in the control group. All mean (SD) scores for DASS-Depression subscales were slightly reduced after the intervention program.In this intervention program, the experimental group received a short duration SMT program  that shows improvement in the mean (SD) scores for DASS-Depression and DASS-Anxiety but not for DASS-Stress. However, when we examined the mean (SD) scores for DASS-Stress (pre and post-intervention), the mean (SD) scores were slightly reduced after the intervention program. In an effort to identify which subscales of DASS-Depression, DASS-Anxiety and DASS-Stress are significantly contributing towards improvement in the mean (SD) scores, we also analysed the effects of the intervention program on each subscale of the DASS-Depression, DASS-Anxiety and DASS-Stress scales. With regards to DASS-Depression subscales, we found that the mean (SD) score for dysphoria (p = 0.007) and situational anxiety (p = 0.048) were significantly reduced after intervention. This suggests that subjective anxiety, as explained by "I had a feeling of faintness" and "I felt scared without any good reason" and situational anxiety as explained by "I found myself in situations that made me so anxious I was most relieved when they ended" are statistically significantly reduced. Meanwhile, the mean (SD) scores for self-perceived DASS-Anxiety subscales, such as skeletal muscle effect (p = 0.495) and autonomic arousal (p = 0.054) were not significantly different pre- and post-intervention. However, when we examined the mean (SD) scores for pre- and post-intervention, all the mean (SD) scores for DASS-Anxiety subscales were also reduced.With regards to the mean (SD) scores for DASS-Anxiety subscales, we found that the mean (SD) scores for subjective anxiety (p = 0.018) and easily upset  (p = 0.047) were significantly reduced after intervention.Whereas for DASS-Stress subscales, we found that the mean (SD) scores for nervous arousal ("I felt that I was using a lot of nervous energy") . Results indicated that SMT is effective in reducing some aspects of self-perceived depression, anxiety, and stress in these workers. Therefore, intervention programs to reduce stress, anxiety, and depression in the workplace may facilitate automotive workers to provide high-quality service in an automotive assembly plant setting.The authors declare that they have no competing interests.RBN, EBA and LN contributed equally to the design and conduct of the survey, analysis of the results, drafting and critical revision of the manuscript. RBN, EBA and LN read and approved the final version of the manuscript


Taurine (T) was first noted as beneficial for stroke and cardiovascular diseases (CVD) prevention in genetic rat models, stroke-prone spontaneously hypertensive rats (SHRSP). The preventive mechanisms of T were ascribed to sympathetic modulation for reducing blood pressure (BP) and anti-inflammatory action. Recent epidemiological surveys revealed the involvement of inflammatory mediators in the pathogenesis of stroke and also atherosclerosis for which T was proven to be effective experimentally. Arterio-lipidosis prone rats, a substrain of SHRSP selectively bred for higher reactive hypercholesterolemia, quickly develop not only arterial fat deposition but also fatty liver which could be attenuated by dietary T supplementation. CARDIAC (CVD and Alimentary Comparison) Study was a WHO-coordinated multi-center epidemiological survey on diets and CVD risks and mortalities in 61 populations. Twenty-four-hour urinary (24U) T was inversely related significantly with coronary heart disease mortality. Higher 24U-T excreters had significantly lower body mass index, systolic and diastolic BP, total cholesterol (T-Cho), and atherogenic index (AI: T-Cho/high density lipoprotein-cholesterol) than lower T excreters. T effects on CVD risks were intensified in individuals whose 24U-T and -magnesium (M) excretions were higher. Furthermore, higher Na excreters with higher heart rate whose BP were significantly higher than those with lower heart rate were divided into two groups by the mean of 24U-T, high and low T excreters. Since the former showed significantly lower BP than the latter, T may beneficially affect salt-sensitive BP rise. Included among the typical 61 populations, were Guiyang, China or St. John’s, Newfoundland, Canada where in which the means of both 24U-T and -M were high or low, respectively. The former and the latter had low and high CVD risks, respectively. Australian Aboriginals living at the coastal area in Victoria were supposed to eat T- and M-rich bush and sea foods and be free from CVD 200 years ago, but they presently have nearly the highest CVD risks indicating that T- and/or M-containing seafood, vegetables, fruits, nuts, milk, etc, similar to prehistoric hunters’ and gatherers’ food should be good for CVD prevention. The preventive effects of T, good for health and longevity, first noted experimentally, were also proven epidemiologically in humans. The health effects of taurine (T), “a wonder molecule”, was first noted in our genetic rat models for hypertension (Spontaneously Hypertensive Rats: SHR) ,2 and stSince SHRSP was established as a genetic model developing stroke, extensive nutritional and pharmacological studies have been conducted experimentally ,9,10,15.Therefore, we further analyzed the nutritional mechanisms of stroke in SHRSP and also in humans. The common predilection site of stroke is the basal ganglia which are fed through perforating arteries branching recurrently from the cerebro-basal arteries. Blood flow through such recurrent arteries is reduced hemorheologically in hypertension, and thus vascular damage develops due to the reduced blood supply to these arterial walls ,16. AnatAfter our demonstration of the importance of inflammation in stroke in rat models, the pathogenic concept of stroke as well as atherosclerosis in humans has also changed greatly . High seBy our further effort for establishing a better model of atherosclerosis from SHRSP, the selective sib-mating of the substrain of SHRSP, developing greater reactive hypercholesterolemia in response to high fat cholesterol (HFC) diet feeding, was started in 1972 . The strDietary T supplementation was proven to be effective for attenuating the development of hypercholesterolemia and for decreasing fat deposits in the mesenteric arteries of the selected SHRSP fed on HFC diets ,27. SuchWe are presently interested in the activation of LOX-1, a receptor for oxidized LDL that was up-regulated in SHRSP fed on HFC diet . T actuaIn relation to arterial lipid deposition we could establish a new model for NASH , now incIn conclusion, our experimental studies consistently indicate greater prospective for T to contribute to the nutritional prevention of CHD, as well as stroke and probably NASH, the increasing health problem of mankind exposed to urbanization and westernization of dietary habit and life styles.These experimental findings to support the beneficial effect of T and other nutrients on CVD prevention led Y. Yamori to propose an international cooperative study on the association of dietary factors with CVD risks and CVD mortality in 1982. The protocol of this study obtained an international consensus of two meetings organized by WHO-Collaborating Center for Primary Prevention of CVD in 1983 and 1985 . The CARIn CARDIAC Study populations we examined 5 diet-related markers in relation to CHD by applying the structural equation modeling for males and females. Five markers were T-Cho, BMI and Na/Cr, M/Cr and T/Cr ratios in 24U , both SBP and DBP, T-Cho and atherogenic index (AI: T-Cho/HDL), an indicator of the risk for developing atherosclerosis and CHD, than low T eaters . SimilarSince the differences in CVD risks between high T-M excreters and low excreters were statistically so high, we further analyzed the additional adverse effect of high salt (Na) intake on low T-M excreters or the additional beneficial effect of low Na intake on high T-M excreters Figure . High anThe population averages of both SBP and DBP in CARDIAC Study showed significantly positive association with the averages of 24U-Na excretion in males but not in females in general. The association was significant only in SBP of postmenopausal women, indicating gender difference in salt-sensitive BP rise . Since gHeart rate (HR) is related to CVD risk in CARDIAC data  as well Since the individuals with greater T and M intakes estimated from 24U excretions had significantly lower CVD risks than lesser T and M excreters , typicalWe are still continuing our CARDIAC health examination since we started the world wide study in 1985 in coordination with WHO. Our recent study on Australian Aboriginal people for over past 7 years demonstrated that they are presently suffering most severely from lifestyle-related diseases in comparison with the other CARDIAC populations ,45. ObesMoreover, they had no salt at all before Captain Cock reached Australia at the beginning of 18th century. Our CARDIAC data help us to speculate how Aboriginals should have been free from CVD risks, based on our comparison of CARDIAC Study individuals consuming a high amount of T and M but less Na with those consuming less T and M but a great amount of Na. CVD risks were clearly and highly significantly lower in the CARDIAC Study participants taking a high amount of T and M but less Na, corresponding to the nutritional balance of Aboriginal ancestors eating traditional bush food. High T and M but less Na-containing foods were supposed to be taken commonly by the ancestors of humans living on hunter-gatherers’ foods like Aboriginal ancestors for nearly 10 times longer than the period after the start of agriculture and farming. Therefore, these dietary conditions seem to be more suitable for the human genome to live a healthy life without CVD.Both T and M are basically important for the maintenance of life. T is involved in cellular physiology by its effect on osmoregulation, anti-oxidant, membrane stabilization and calcium regulation -49, and Moreover, in relation to the ageing of vascular tissue and the accelerated senescence of endothelial progenitor cell (EPC) noted in SHRSP , T was rOn the other hand, M is the 8th abundant element in the weight, the number of atoms and the volume percentage of all atoms on the earth. It is also the most abundant intracellular divalent cation and is involved in the various biological functions of about 300 enzymes as their coenzyme. They include all enzymatic reactions requiring ATP, such as Na-K ATPase, important for intracellular ionic balance. Therefore, M is supposed to be causatively and clinically related to cardiovascular health, hypertension and diabetes -58. LoweSince the evolutional origin of life of human beings was inside the sea containing abundant M and food gatherers lived on seafood rich in T, both M and T are assumed to be essential for the homeostasis maintaining cardiovascular health.In addition to the pathophysiological and various physiological functions of T, the preventive effect of T was first experimentally demonstrated in hypertensive rat models, SHR and SHRSP developing hypertension and stroke genetically. World-wide epidemiological studies conducted for the last 25 years revealed that 24U-T was inversely associated with the age-adjusted mortality rates of stroke and CHD. High 24U-T, particularly combined with high M excretion, was associated highly significantly with lower CVD risks, obesity, hypertension, hypercholesterolemia and AI. These findings indicate the consistency of the beneficial T effects on animal models and humans proven either experimentally or epidemiologically. Such consistency of the extensive basic and epidemiological findings of T effects and the effects combined with M indicates greater prospective for T to contribute to the nutritional prevention of CVD and lifestyle-related diseases.AI: atherogenic index, BMI: body mass index, ALR: arterio-lipidosis prone rats, Ca: calcium, CARDIAC Study: Cardiovascular Diseases and Alimentary Comparison study, CHD: coronary heart diseases, Cr: creatinine, CRP: C-reactive protein, CVD: cardiovascular diseases, DBP: diastolic blood pressure, ER: endoplasmic reticulum, HDL: high density lipoprotein, HR: heart rate, K: potassium, LDL: low density lipoprotein, M: magnesium, Na: sodium, NASH: nonalcoholic steato-hepatitis, SBP: systolic blood pressure, SHR: spontaneously hypertension rats, SHRSP: stroke-prone spontaneously hypertension rats, T: taurine, T-Cho: total cholesterol, 24U: 24-hour urine.The authors declare that they have no competing interests.YY designed taurine-related studies and prepared for this review. MM and HM conducted epidemiological surveys. TT contributed to the data analysis. AM conducted laboratory analyses. KK helped experimental studies and reference collection.


To better understand income-related inequalities in health care use, it is imperative to identify sources of inequalities and assess the extent to which health care use is still related to income after differences in need across the income distribution are accounted for. Little is known regarding rural-urban differences in income-related inequalities and subgroup variation in horizontal inequities in health care use. This study decomposes income-related inequalities in ambulatory care use into contributions of need and non-need factors and compares horizontal inequities of subgroups in rural and non-rural areas.This analysis used non-elderly adult samples from the 1998 to 2001 U.S. National Health Interview Survey data. The area of residence was categorized as rural for non-Metropolitan Statistical Area (MSA) and non-rural for MSA. Concentration indices of ambulatory care use were used to gauge income-related inequalities and decomposed into contributing factors. Horizontal inequities were measured using two methods and the results were compared.Ambulatory care use was disproportionately concentrated in the poor before need adjustment. However, the results of decomposition and horizontal inequity analyses indicate that the pro-poor concentration of health care use was due to greater health care need in low-income groups. Adjusting for need, ambulatory care use was distributed favoring the better-off, to a larger degree in non-rural areas. Health-related variables were the major contributors to income-related inequalities. Non-need factors, including socioeconomic factors, health insurance, and usual source of care, also contributed to income-related inequalities. There were variation in determinants' contributions to income-related inequalities between rural and non-rural populations and subgroup differences in horizontal inequities. Horizontal inequities were greater within non-whites, high school graduates, individuals with private health insurance, and those without a usual source of care with some geographic variation.Our analysis shows that seemingly pro-poor income-related inequalities in ambulatory care use were largely due to greater health care need among low-income groups. The results demonstrate different contributions of determinants to income-related inequalities and variation in horizontal inequities by subgroup and locale. The findings of this study should help identify targets for policy intervention for each rural and non-rural area. Income inequalities have been associated with differences in health outcomes including mortality, mental and physical health, and reproductive outcomes -5. ReseaHealth inequalities are partly attributed to unequal health care use across sociodemographic groups ,15. StudHorizontal equity is a widely accepted concept in health inequality research ,19. The HIwv), a measure of inequity in health care use, can be obtained either (1) by calculating the concentration index of need-standardized health care use or (2) by subtracting the concentration index of need-predicted (or need-expected) use (CN) from that of actual care use (CU) [Horizontal inequity index (use (CU) -24. Sincuse (CU) . Due to HIwv indicates a higher share of health care use of higher-income groups than their share of need, indicating horizontal inequities favoring the better-off. On the contrary, a negative value of HIwv represents horizontal inequities favoring the poor given their share of need [A negative value of a concentration index signifies health care need/use favoring lower-income groups. When the concentration index is positive, health care need/use is more concentrated in higher-income groups. If the concentration index is zero, health care need/use is distributed equally regardless of income level . A posit of need .When individual-level information is available from microdata , a horizontal inequity index can be obtained based on the estimates from indirect need standardization, in which non-need variables in addition to need variables are included in regression ,24. HereCU - CN), the concentration index for need-predicted health care use should be estimated first [CU) is calculated based on the observed number of health care use .To compute a horizontal inequity index using the second method  decompose income-related inequalities in health care use by source of the inequalities among U.S. non-elderly population; 2) measure degrees of horizontal inequities adjusting for differences in health care need across the income distribution; and 3) compare income-related inequalities, determinants of inequalities, and horizontal inequities by rural-urban locale.This analysis used pooled data of non-elderly adult samples aged 18 to 64 from the 1998, 1999, 2000, and 2001 U.S. National Health Interview Survey (NHIS). NHIS is an annual survey of a nationally representative household sample using random-digit dialed telephone interviews to collect information on a broad range of health topics including illness, disability, chronic impairments, and health care use of noninstitutionalized civilian population of the 50 states and the District of Columbia . The areThe dependent variable is the frequency of ambulatory health care use as measured by the number of health professional contacts including visits to doctors' office, telephone contacts, and home care services during the past two weeks. Health professional refers to physicians, nurses, and other health care professionals such as physical therapists, ophthalmologists, and chiropractors .Equivalent individual income was calculated based on reported family income using the modified Organization for Economic Co-operation and Development (OECD) equation formula. Equivalent individual income is defined as the income of an individual adjusted for differences in family size and composition. The equivalence scale formula is:Family income included all sources  from all family members who were 18 years or older.CN), the contribution of the following need variables was included: 6 categories of age-sex variables , 4 dummy variables derived from a five-point Likert-type scale of self-assessed health status , and dummy variables for activity limitations and illnesses. Activity limitation was coded 1 if a respondent reported any activity limitation due to mental, physical, or emotional problems and 0 otherwise. Illness was also coded 1 if a respondent reported any illness condition and 0 otherwise.In computing the concentration index of need-predicted use  is as follows:In statistical analyses, steps were taken to correct errors occurring in pooled data with different sampling weights by year. Fahimi's weighting method for pooled data  was usedk, is the number of surveys to be pooled, ki Wis the sampling weight for the i th sample of k survey datasets. The sample size of pooled data is n = n1 + n2 + ......... + kn. To avoid inflated variance in pooled data, weights (kiW) from each sample have to be normalized with respect to their corresponding sample sizes [where N is the population size, le sizes . For prele sizes  using thle sizes .A concentration index measuring income-related inequalities in health care use was calculated using a convenient regression method as follows:i, and μ is the mean of health care use yi, and ri is the weighted fractional rank of utilized income, defined aswhere i is the sample weight scaled to sum to 1 [where wsum to 1 . The conThe concentration index was decomposed into the contribution of individual factors to income-related inequalities following the method proposed by van Doorslaer, Koolman, and Jones . For nonαm is an intercept, i εis an error term, and δm, βmk and γmj are the partial effect of log income (ln inc), a set of k need and j non-need variables, respectively. Given the relationship between yi and xki, the overall inequalities in health care use C is expressed aswhere μ is the mean of y, k Cand j Care the concentration index of k xand jz, and ε GCis the generalized concentration index of the error tem ε. The overall inequality in an outcome has two components: an explained component and an unexplained component. The residual component reflects the income-related inequality that is not explained by systematic variation of the determinants in the model. The explained component has two main elements: the impact of each determinant on an outcome and the extent of unequal distribution of each determinant across income groups [where e groups .CN) from that of actual health care use (CU) as follows [The extent of horizontal inequity was measured by the concentration index of need-standardized health care use and by subtracting the concentration index of need-predicted use . Horizontal inequity indices for subgroups  were also calculated to examine whether the degree of horizontal inequity varies across the subgroups. In calculating horizontal inequity indices for subgroups, all independent variables except for the group variable of interest were included in need standardization. For example, in measuring the horizontal inequity indices of white and non-white subgroups, all independent (need and non-need) variables were included in need standardization except for the race indicator variable. All analyses were performed with the whole sample and with rural and non-rural samples separately.where Table Ambulatory care use was disproportionately concentrated in the poor before need adjustment. The crude concentration indices for the whole sample and rural and non-rural subsamples were negative, indicating income-related inequalities in health care use favoring the poor when health care need was ignored. The negative values of need-predicted concentration indices  signify that expected health care use given health care need is larger among the poor than among the better-off. Overall, the distribution of health care use given health care need was in favor of the better-off as indicated by the positive value of the horizontal inequity index (0.118). Each rural and non-rural population also showed pro-rich horizontal inequity in ambulatory care use.Decomposition analyses illustrate the sources of pro-poor income-related inequalities in ambulatory care use. Table The associations between determinants and the outcome were in expected directions. Having health-related problems (activity limitation and illness), perceiving one's health not excellent, and having higher education, higher income, and public insurance were associated with more ambulatory care use when all other factors were equal. Being non-white or less educated and having no insurance or usual source of care were associated with less ambulatory care use in both rural and non-rural populations when all other factors were equal.Decomposition of concentration indices also showed expected distribution of determinants in income groups. Positive (negative) concentration indices indicate concentration of individuals with specific characteristics in higher (lower) income groups. For example, individuals with above-high-school education were concentrated in higher-income groups . In contrast, individuals with below-high-school education were disproportionately concentrated in lower-income groups . Non-whites, people with public insurance, people without health insurance or a usual source of care, and those in poor health were disproportionately concentrated in low-income groups.The last column for each locale in Table For both rural and non-rural populations, health-related variables  as a group were the largest contributor to income-related inequalities because of their unequal distribution in low-income groups and strong positive association with health care use. Activity limitation was the single largest contributor to income-related inequalities as indicated by 57% in rural and 152% in non-rural areas. It can be interpreted that the total inequalities in health care use favoring the poor, with all else equal, would have been 57% lower in rural areas if individuals with activity limitations were equally distributed across income groups or if having activity limitation was not associated with health care use as indicated by the regression coefficient (0.355) in the model. Both SES variables (education levels and income) showed negative inequality contributions. For example, due to their concentration in low-income (high-income) groups and negative (positive) association with health care use, both education groups (below-high and above-high school groups) lowered the pro-poor income-related inequalities. The negative inequality contribution of being uninsured was particularly strong in non-rural areas. In other words, the higher concentration of the uninsured in low-income groups and their lower number of ambulatory care use substantially decreased the extent of pro-poor income-related inequalities in health care use in non-rural areas.The results of horizontal inequity analyses indicate that the disproportionate concentration of health care use in low-income groups was due to their greater health care need. When need was accounted for, ambulatory care use was actually distributed favoring high-income groups.CN from CU   were similar, but not identical to the concentration indices computed by subtracting Horizontal inequity can also be assessed based on the regression coefficients from decomposition analyses Table . AssuminTable HIwv = 0.089 vs. 0.065). Education seems to significantly reduce horizontal inequities in non-rural areas, where the high-school graduate group had greater horizontal inequity (HIwv = 0.055) than the group with higher education (HIwv = 0.024). In rural areas, examination of horizontal inequity by insurance coverage showed pro-rich horizontal inequities only among residents with private health insurance. In non-rural areas, horizontal inequities were significant among both privately and publicly insured groups. Across locales, having a usual source of care or not appears to be closely linked to the extent of horizontal inequity. Rural residents without a usual source of care experienced significant horizontal inequity (HIwv = 0.123). In non-rural areas, the pro-rich horizontal inequity was much greater among residents without a usual source of care (HIwv = 0.151) than among those with (HIwv = 0.074).Horizontal inequity in health care use was present within both whites and non-whites in non-rural areas, but was more substantial among non-whites  and higher income and not having a usual source of care decreased the pro-poor income-related inequalities because of these determinants' associations with health care use and concentration in the income distribution. Horizontal inequity indices indicate health care use favoring the better-off in both rural and non-rural areas when health care need was accounted for. However, the magnitude of horizontal inequity was greater in non-rural areas. Racial minorities, high school graduates, those with private health insurance, and people lacking a usual source of care were likely to experience relatively higher degrees of within-group horizontal inequities than their respective counterparts with some geographic variation.The findings of this study provide information for policy makers to reduce inequalities in health care use and, in turn, health disparities. Different contributions of determinants to income-related inequalities in health care use and variation in horizontal inequities by subgroup and locale point to more explicit targets for policy amendment and resource allocation for each rural and non-rural area.The authors declare that they have no competing interests.HS analyzed data, interpreted the results, and drafted the initial version of the manuscript. JK interpreted the results and wrote the final version of the manuscript. All authors read and approved the final manuscript.


The majority of cancer patients experience dramatic weight loss, due to cachexia and consisting of skeletal muscle and fat tissue wasting. Cachexia is a negative prognostic factor, interferes with therapy and worsens the patients' quality of life by affecting muscle function. Mice bearing ectopically-implanted C26 colon carcinoma are widely used as an experimental model of cancer cachexia. As part of the search for novel clinical and basic research applications for this experimental model, we characterized novel cellular and molecular features of C26-bearing mice.A fragment of C26 tumor was subcutaneously grafted in isogenic BALB/c mice. The mass growth and proliferation rate of the tumor were analyzed. Histological and cytofluorometric analyses were used to assess cell death, ploidy and differentiation of the tumor cells. The main features of skeletal muscle atrophy, which were highlighted by immunohistochemical and electron microscopy analyses, correlated with biochemical alterations. Muscle force and resistance to fatigue were measured and analyzed as major functional deficits of the cachectic musculature.We found that the C26 tumor, ectopically implanted in mice, is an undifferentiated carcinoma, which should be referred to as such and not as adenocarcinoma, a common misconception. The C26 tumor displays aneuploidy and histological features typical of transformed cells, incorporates BrdU and induces severe weight loss in the host, which is largely caused by muscle wasting. The latter appears to be due to proteasome-mediated protein degradation, which disrupts the sarcomeric structure and muscle fiber-extracellular matrix interactions. A pivotal functional deficit of cachectic muscle consists in increased fatigability, while the reported loss of tetanic force is not statistically significant following normalization for decreased muscle fiber size.We conclude, on the basis of the definition of cachexia, that ectopically-implanted C26 carcinoma represents a well standardized experimental model for research on cancer cachexia. We wish to point out that scientists using the C26 model to study cancer and those using the same model to study cachexia may be unaware of each other's works because they use different keywords; we present strategies to eliminate this gap and discuss the benefits of such an exchange of knowledge. Cancer genome projects are providing complete landscapes of the mutations that exist in tumors, making it essential to bridge the gap between high-throughput sequencing information and the molecular mechanisms underlying the natural history of cancer . In thisCancer cachexia is a muscle wasting syndrome that affects most cancer patients . Cachexiin vivo by inoculation of serial cell doses into syngeneic BALB/c mice. The C26 line was highly tumorigenic and displayed a low tendency to metastasize; C26-inoculated mice exhibited high mortality  a complex metabolic syndrome associated with underlying illness and characterized by loss of muscle with or without loss of fat mass [...]" - translaTibialis Anterior; TUNEL: Terminal dUTP nick end labeling; WB: Western Blot.BrdU: 5-bromo-2'-deoxyuridine; C26: colon carcinoma C26; CSA: Cross Sectional Area; EDL: Extensor Digitorum Longus; CXCR3: CXC Chemokine Receptor 3; H&E: Hematoxylin and Eosin; IL-6: interleukin-6; MAP: Mitogen-Activated Protein; MMP: Matrix MetalloProteinase; NADH: Nicotinamide Adenine Dinucleotide: reduced; Peg3/PW1: Paternally Expressed Gene 3; PI: propidium iodide; RT-PCR: Reverse-Transcriptase Polymerase Chain Reaction; SEM: Standard Error of the Mean; TA: The authors declare that they have no competing interests.PA, EB and VC, conception, design and execution of experiments; ER, physiology experiments; BP, contributing important intellectual content; CR and FP, data collection; EPS, AB and FF, histopathology of the tumor and major intellectual contribution; SA, concept, design and editorial support, research fund collection; DC, conception and design, data collection and analysis, figure and manuscript preparation. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2407/10/363/prepub


Helicobacter pylori and the gastric MALT lymphoma is a well known issue, but recently there are several data suggesting the role of hepatitis C virus (HCV) infection in the pathogenesis of lymphoma including MALT lymphoma. Herein we present a rare case of duodenal MALT lymphoma with multiple intra-abdominal thromboses together with HCV infection that was confirmed by real-time polymerase chain reaction detecting HCV-RNA within the peripheral blood mononuclear cells.Mucosa associated lymphoid tissue MALT lymphoma is a low grade malignancy that arises most commonly from the gastric mucosa. Small intestinal involvement is very rare. The causative relationship between  Helicobacter pylori in the pathogenesis of gastric MALT lymphoma is a well known issue. On the other hand, recently there are some clues that hepatitis C virus (HCV) infection may be a possible contributing infectious factor in the pathogenesis of MALT lymphoma [Mucosa associated lymphoid tissue (MALT) lymphoma is a neoplasm with low grade malignancy, arising from MALT of various organs including gastrointestinal system, salivary glands, lungs, thyroid glands, thymus, mammary glands and lacrimal glands . Among tlymphoma ,3.Simultaneous portal, splenic and mesenteric vein thrombosis is a rare condition and may be associated with malignancy, surgery, portal hypertension, hyperhomocysteinemia, Behçet's disease, oral contraceptive use, factor V Leiden mutation, local inflammatory conditions and hematological hypercoagulable states such as paroxysmal nocturnal hemoglobinuria and myeloproliferative disorders -6.Herein we present a case with a history of periodic abdominal pain bouts who admitted with new onset of severe abdominal pain, ascites, signs of malabsorption, and multiple intraabdominal venous thromboses.A 32 year old male presented with severe abdominal pain, fatigue and nausea. He had a seven years history of periodic abdominal pain attacks unrelated to meals and lasting for several days. He had once appendectomy and once laparotomy for acute abdominal findings. Because he had recurrent bouts of abdominal pain, colchicine treatment was started considering the diagnosis of familial Mediterranean fever (FMF). No objective improvement was achieved on this treatment and the patient stopped taking the drug because of the adverse effects such as diarrhea. For the last three weeks, the abdominal pain persisted, became very severe and worsened after meals with nausea and vomiting. In physical examination, the patient was pale and cachectic, abdominal examination revealed guarding and diffuse abdominal tenderness to palpation. There was ascites at level of umbilicus. The patient's past history and family history were noncontributory. In the initial radiological and biochemical assessment, there were no sign of acute abdomen and thus laparotomy was not indicated.Results of initial laboratory analyses are shown in table In abdominal ultrasonography (USG), ascites and portal and splenic vein thromboses were observed. Abdominal computed tomography (CT) and CT angiography showed increased wall thickness of intestinal loops, splenic, portal and superior mesenteric vein thrombosis. Arterial pathology was not detected Figure . In smalTo determine the cause of multiple venous thrombosis in the abdomen, thrombophilic parameters were investigated: Anti-cardiolipin immunoglobulin (IG) G: 4.6 GPLU/mL; anti-cardiolipin IG M: 36.3 MPLU/mL; homocystein (fasting level): 25.1 μmol/L ; protein C: 94%; protein S: 83%; lupus anticoagulant was negative; factor V Leiden and prothrombin gene mutations were not detected. Immunophenotyping in the peripheral blood revealed normal CD55 and CD59 expression and paroxysmal nocturnal hemoglobinuria (PNH) was not considered. Vasculitis might be responsible for the thrombosis, so rheumatologic diseases were investigated. For Behçet's disease, pathergy test, HLA-B51, HLA-B52 were all negative. Anti-DNA, anti nuclear antibody, anti neutrophil cytoplasmic antibody, anti- Jo-1, anti-RNP, anti-Scl-70, anti-Sm, anti SS-A, anti- SS-B and rheumatoid factor were negative. FMF was considered because of the recurrent bouts of abdominal pain, but available FMF mutation assays were negative. Viral serology was negative for hepatitis B virus (HBV) surface antigen, anti HBV surface antibody and anti-HIV. Anti-HCV (ELISA) was positive but HCV-RNA was negative . To investigate possible cryoglobulinemia associated with hepatitis C infection; cryoglobulin and cryofibrinogen tests were performed and found negative.Because the patient had anti-HCV positivity but HCV-RNA negativity with normal liver transaminases, to confirm the hepatitis C virus infection, HCV-RNA in the plasma and peripheral blood mononuclear cells (PBMC) was tested by RT-PCR method . HCV-RNA was detected in peripheral blood mononuclear cells, so the patient was considered to have hepatitis C virus infection. Liver biopsy revealed non-specific reactive hepatitis and no sign of chronic viral hepatitis, so antiviral treatment was not started.Malabsorption associated with abdominal pain raised the suspicion of intestinal T cell lymphoma following celiac disease. Anti-gliadin immunoglobulin (Ig) A was 1.2 U/mL and anti-gliadin Ig G was 20.4 U/mL. Anti-endomysium Ig A was negative and serum Ig A level was 163 mg/dL , so diagnosis of Celiac disease was not considered.Helicobacter pylori was negative in biopsy specimens. Colonoscopy was also performed and was normal. In bone marrow biopsy, mild hypercellularity was seen and infiltration was not detected.In gastroduodenoscopy, duodenum was observed edematous, fragile and granular; multiple small ulcerated lesions were seen Figure . Second 2 and 2 mg vincristine for once and 100 mg/day prednisolone for 5 days) was started. Because Helicobacter pylori was negative in pathology specimens, eradication therapy was not administrated. At follow up, HCV RNA was still negative and liver function tests remained stable. Because the patient had multiple venous thromboses in abdomen, enoxaparine in treatment dose followed by warfarin was started. After these treatments, patient's complaints and findings of malabsorption were dramatically improved. Control gastroduodenoscopy 4 months after treatment revealed regression of the lesions; endoscopic biopsies were repeated in which no infiltration was detected. Follow up small intestinal passage radiograph was normal.Diagnosis of MALT lymphoma of the small intestine was considered and COP treatment protocol  lymphoma accounts for approximately 30% of all extranodal lymphomas ,7. HowevThe presenting symptoms of duodenal lymphoma are insidious and non-specific and depend on the location of the tumor, the degree of obstruction, and the rapidity of growth. Our case presented with abdominal pain, nausea and vomiting especially just after the meals probably related to intestinal obstruction. He also had signs of malabsorption due to infiltration of the duodenum and jejunum. After successful treatment of lymphoma and nutritional replacement therapy, all these findings subsided.Helicobacter pylori infection and possibly HCV infection. Infectious agents like Helicobacter pylori, Campylobacter jejuni, Borrelia burgdorferi, Chlamydia psittaci and HCV indirectly increases the probability of lymphoid transformation by chronically stimulating the immune system to maintain a protracted proliferative state [Predisposing factors for lymphoma of the small intestine include prior malabsorption syndromes (e.g. Celiac sprue and dermatitis herpetiformis); inflammatory bowel disease (e.g. Crohn's disease), acquired immunodeficiency states, ve state .Helicobacter pylori, as another potential infectious co-factor in the multiple-stage pathogenesis of low-grade lymphomas of MALT type [HCV is both hepatotropic and lymphotropic. Association between HCV infection and mixed cryoglobulinemia, a benign lymphoproliferative disorder is well known but the role of HCV in the pathogenesis of frank lymphoma is not as certain, and is under ongoing discussion, but some authors suggest that HCV may be considered, in addition to ALT type .Isolated anti-HCV positivity in individuals with negative serum HCV-RNA may be related to past infection or false positivity, but this has not yet been completely understood. In our case, anti-HCV was positive but HCV-RNA was negative and liver enzymes were normal. To confirm the diagnosis of hepatitis C virus infection, HCV-RNA was detected by RT-PCR within the PBMCs. Presence of HCV within the lymphoid cells may have contributed to development of intestinal MALT lymphoma, but to our knowledge there is no published report investigating the association of isolated anti-HCV positive hepatitis C virus infection and lymphoma.After the chemotherapy including corticosteroids, our patient was followed by serial liver enzymes and HCV-RNA tests, but signs of activation of HCV infection did not occur; liver enzymes were always in normal range and HCV-RNA was persistently negative.One of the unique features of this case is the associated multiple intraabdominal venous thrombosis, including portal vein, splenic vein and superior mesenteric vein. Patients with combination of these thromboses may present with abdominal pain or findings of chronic portal hypertension ,6. Our pOne of the most important mechanisms of hypercoagulability in malignant diseases is the action of tissue factor, which may be tumor cell-derived  or may oHelicobacter pylori and antiviral treatment for HCV may cause regression of the MALT lymphoma and may be used for its treatment similar to Helicobacter pylori eradication therapy. Pathophysiological link between HCV infection and MALT lymphoma and its impact on the treatment should be further investigated.We have reported a rare case of MALT lymphoma of the duodenum and jejunum together with multiple intraabdominal thromboses and HCV infection proven by detection of HCV-RNA in PBMCs. All patients with MALT lymphoma should be screened for HCV infection. Isolated anti-HCV positivity should be confirmed by detection of HCV-RNA in PBMCs to exclude false positivity, because diagnosis of HCV infection is important in follow up of patients who are given immunosuppresive chemotherapeutics for lymphoma. HCV infection may play a role in the pathogenesis of MALT lymphoma beside Written informed consent was obtained from the patient for publication of this case report and accompanying images. A copy of the written consent is available for review by the journal's Editor-in-Chief.The authors declare that they have no competing interests.AO: Follow-up of the patient and writing the manuscript.FT: Follow-up of the patient and writing the manuscript.SN: Follow-up of the patient and writing the manuscript.MB: Forming and explanation of the radiological figures.BP: Providing the explanation of the endoscopic findings.OD: Forming and explanation of the pathology figures.MAK: Follow-up of the patient and writing the manuscript.CT: Follow-up of the patient and writing the manuscript.All authors read and approved the final manuscript.


Bacillus anthracis and B. cereus, but has gone unrecognized because it is always encoded far from its maturation protein operon. Patterns of insertions and deletions among twenty-four variants suggest a repeating functional unit of Cys-Xaa-Xaa.Bacteriocins are peptide antibiotics from ribosomally translated precursors, produced by bacteria often through extensive post-translational modification. Minimal sequence conservation, short gene lengths, and low complexity sequence can hinder bacteriocin identification, even during gene calling, so they are often discovered by proximity to accessory genes encoding maturation, immunity, and export functions. This work reports a new subfamily of putative thiazole-containing heterocyclic bacteriocins. It appears universal in all strains of This article was reviewed by Andrei Osterman and Lakshminarayan Iyer. The N-terminal region is a leader peptide that may show homology from one precursor to another. The C-terminal region, however, contains low-complexity sequence, often repetitive and rich in Cys, Ser, or Thr, that may be hard to distinguish from spurious translations of miscalled genes. Second, a putative cyclodehydratase encoded by a neighboring gene joins the side chain of a Cys, Ser, or Thr residue to the carbonyl group contributed by the immediately N-terminally adjacent residue, with the elimination of a water molecule, to form a heterocyclic ring. Third, a dehydrogenase is encoded in the cluster. The dehydrogenase acts on the resulting thiazoline (Cys) or oxazoline (Ser or Thr) ring to produce a thiazole or oxazole ring. Lastly, a protein described as a docking scaffold appears in the same cluster. Additional functionally connected genes may co-occur in these clusters, including putative transporter subunits, immunity proteins, proteases, and enzymes for other types of modifications, but none is required.Many bacteria produce bacteriocins, toxins built by post-translational modification of ribosomally produced precursors . RecentlWe developed hidden Markov model (HMM) search models  for the Bacillus anthracis [Bacillus, with no candidate bacteriocin precursor gene nearby. This observation suggested that the toxin precursor gene might be present in the genome but at some distance from the maturation cluster.In the genome of nthracis , the caunthracis  with a fnthracis , as wellB. anthracis Ames, BA_2677 (GB|AAP26522), with sequence features appropriate for a heterocycle-containing bacteriocin precursor. In particular, the C-terminal region has an exaggeratedly low amino acid complexity. It is Cys-rich, Gly-rich, and highly repetitive; the thirteen Cys residues are regularly interspaced with either two amino acids (ten times) or three amino acids (twice).At a distance of 1.27 megabases from the cluster, we identified an 88-residue protein in Bradyrhizobium japonicum. We constructed HMMs from alignments of constructed Cys-rich repetitive sequences with periodicities of two (Cys-Xaa) or three (Xaa-Cys-Xaa), repeated four to eight times. The default behavior of the hmmbuild program in HMMER 2.0 is to detect largely redundant sequences and replace actual counts of the different residues seen at each position by "effective counts." Overriding this behavior with the --noeff option forces the use of actual counts rather than effective counts in Cys-only columns of the alignment, sharpening the distinction between Cys itself and all other amino acids. The resulting series of HMMs, based on from four to eight repeats of the tripeptide sequence Xaa-Cys-Xaa, consistently reported BA_2677 as the top or only match in the genome. Searches based on repeated Cys-containing dipeptide repeats, as in the Staphylococcus aureus RF122 bacteriocin precursor YP_416854, found no candidate bacteriocin in B. anthracis.Only a fraction of the putative thiazole-containing bacteriocins put forward by Lee, et al. have extended Cys-rich repetitive regions, but such regions are highly suggestive in those that do, such as the Cys-Arg-Gly repeats in a precursor from B. anthracis as a spore-forming lethal pathogen with biowarfare capability, and the medical significance of the B. cereus group in both infection and food poisoning [B. anthracis, B. cereus, and other Bacillus species. Therefore, numerous putative orthologs could be found in completed genomes in the genus Bacillus. In each case, a member was found only if and only if orthologs to BA_1266 (dehydrogenase), BA_1267 (scaffolding protein), and BA_1268 (cyclodehydratase) were present in the same genome. In Bacillus cereus ATCC 14579, with two such sets of maturation proteins, two putative protoxins were found. More commonly, however, protoxins appear to be accommodated by a single maturation protein cassette, consistent with the broad specificities inferred from heterologous expression studies.The high interest in oisoning , have heBacillus licheniformis ATCC 14580. A non-overlapping region of the same open reading frame represents the best match in the genome to HMMs based on Cys-Xaa-Xaa repeats. The respective candidate bacteriocins from B. anthracis and B. licheniformis are similar in Cys content and spacing, if not in their interleaving sequence residues; triplets of Cys-Gly-Gly in B. anthracis are largely replaced by triplets of Cys-Trp-Ser in B. licheniformis. Interestingly, the tetrapeptide Cys-Trp-Ser-Cys also occurs in a known bacteriocin, thuricin S [An HMM was built from the N-terminal region of collected BA_2677 homologs in order to represent the presumptive leader sequence. This HMM identified a region of sequence similarity to a previously uncalled putative bacteriocin precursor gene in uricin S , within B. licheniformis gene, however, is adjacent to two appropriate maturation genes, AAU22609 (scaffolding) and AAU22610 (cyclodehydratase). A reciprocal search, initiated from the N-terminal 35 amino acids from B. licheniformis, performed by PSI-BLAST , finds BA_2677 immediately as the top hit . PSI-BLAST converges in the third round to recover 24 non-identical sequences closely related to BA_2677 in the leader sequence region, and it finds no other sequences. These results suggest that BA_2677, and TIGRFAMs model TIGR03601 as an HMM for its detection, represent a new family of heterocycle-containing bacteriocins of the class described. The supporting evidence includes bidirectional best hit matches between the leader sequence regions, similar Cys-rich tripeptide repeat structures in the C-terminal regions, and colocalization of the B. licheniformis member with two distinctive components, the cyclodehydratase and the scaffolding protein, of canonical heterocycle biosynthesis cassettes. Figure The B. anthracis, B. cereus, B. thuringiensis, and B. weihenstephanensis. The most abundant tripeptide was CGG (152), outnumbering its phase-shifted alternatives GGC (95) and GCG (76), as well as RCG (83). The multiple alignment therefore is edited [Figure s edited  in Cys/GThe primary function of most bacteriocins is bacteriocidal activity against closely related species or strains. However, these secondary metabolites may have important roles in pathogenesis, in food preservation, as antibiotics, or as pharmacologically active compounds that may lead to the development of new drugs. Since the maturation proteins of heterocyclic bacteriocin have been shown to act rather freely on protoxins from other bacterial lineages , it is rHMM: hidden Markov model; ORF: open reading frame.The author declares no competing interests.DHH performed the analyses, interpreted the results, and wrote the paper.Andrei Osterman, Burnham Institute for Medical Research, La Jolla, CA.A short article by D. Haft provides us with another beautiful illustration of the power of comparative genomic approach for pathway reconstruction and gene discovery. This study builds on top of a recently published analysis of widespread genomic distribution of pathways generating bacteriocins, short proteins containing oxazole and/or thiazole heterocycles. By combining highly sensitive profile-based search models with the analysis of genome context, D. Haft predicted the existence of a putative bacteriocin maturation pathway in an important group of Bacillus (B. anthracis and B. cereus). The most challenging aspect of this analysis was a "missing" bacteriocin precursor, which in all previously described cases was located in the same chromosomal cluster with genes involved in its posttranslational modification. A convincing prediction of a remotely located gene candidate for a novel version of protoxin characteristic of this group is the most impressive intellectual breakthrough of this elegant study. The identification of an uncalled gene encoding an ortholog of the "heterocycloanthracin" precursor in a chromosomal cluster with the maturation pathway genes in B. licheniformis strongly validated this ambitious inference. A unique and highly attractive feature of D. Haft's article is that it provides biologists with a mature hypothesis that may (and most certainly will) be immediately tested by a focused experiment.Lakshminarayan Iyer, NCBIB. licheniformis gene, that these are bacteriocin genes.The author describes a bacteriocin gene family from Bacillus species that is not in the vicinity of its predicted maturation genes. As bacteriocins are generally short, can be missed during gene annotation, and not very conserved in sequence, isolating them is of some importance. The main point of this study is that while most bacteriocins are encoded next to the genes involved in their maturation, this particular family, barring one exception, appears to be encoded far away from them. I have confirmed the results independently and agree, based on the evidence from the I have a few minor comments.- I recommend adding the source of the sequences in the figure for better clarity. Typically the species names are given as abbreviations.licheniformis bacteriocin as a starting point and the database/s against which it was searched should be mentioned for reproducibility, as a simple web PSI-BLAST search against nr does not yield the result as described in the text. However, upon searching against individual genomes, the bacteriocin sequences are retrievable.- The parameters of the PSI-BLAST search using the predicted The strain of origin for every sequence has been added to the legend for figures Because of the small size of the putative protoxin, especially once repetitive sequence is removed to leave only 35 N-terminal amino acids, PSI-BLAST requires an E-value more permissive than the default value of 10. The proper search parameters are now described explicitly in the text.