Tetanus is a serious but vaccine-preventable disease and fatality rate of the disease is high in the neonates and the elderly. The aim of this study was to detect the tetanus antibody prevalence in the over sixty-year age residents of the nursing homes in Bolu.A voluntary-based study was done in the residents of two nursing homes in Bolu, Turkey. Blood samples were taken from 71 volunteers residing in there nursing homes. Tetanus IgG antibodies were measured by a commercial ELISA kit.Among overall subjects, only 11 (15.7 %) had the protective tetanus antibody titers at the time of the study. Totally, 10 subjects were examined in emergency rooms due to trauma or accidents within the last ten years and, four (40%) of them had protective antibody levels. Of the remaining 61 subjects only 7 (11%) had protective antibody levels (p < 0.05) [Relative Risk = 3.49, 95% Confidence Interval 1.24–9.77].Tetanus antibody level is below the protective level in the majority of the over-sixty-year-age subjects residing in the nursing homes. Each over sixty-year age person in our country should be vaccinated. Until this is accomplished, at least, nursing home residents should be vaccinated during registration. Clostridium tetani. Tetanus spores are present in soil and manure and may be introduced into the body through a puncture wound, burn or scratch. Tetanus can never be eradicated because the spores are widely spread in the environment, and found in the intestinal flora of domestic animals, horses, chickens, and humans. Tetanus is not spread from person to person [Tetanus is an acute disease caused by the tetanus toxin, released by the bacterium o person .Tetanus is preventable by proper immunization. However, the disease is still prevalent even in the most developed countries and it often emerges in the elderly . The preThe mortality rate of tetanus is high in the neonates and the elderly patients . Age-relIn the USA, the disease occurs at a six-times higher rate in this age group ,9. Of 99Bolu is a city in the Western Black Sea Region of Turkey and has a population of approximately 283.000. There are two nursing homes  in the each with a boarding capacity of 60 people. Tetanus antibody level in such a specific group has not been researched in the country yet. The purpose of this study was to detect tetanus antibody prevalence in the voluntary residents of the nursing homes over 60 years old.This study was performed May 1–30, 2004. Subjects were selected from Izzet Baysal and Neziha Baysal Nursing home residents who volunteered for antibody measurement.A standard form was filled including subjects' clinical information, demographic traits along with vaccination and injury stories. 10 cc venous blood was taken from each subject and kept at -20°C until the day of study.Tetanus Ig G antibodies were measured by a commercial ELISA kit [Novatec Dietzenbach, Germany] at a 450/620 nm wavelength. The results were evaluated in the way previously defined by Schoroder et al. . BrieflyStudents't-test was used to compare quantitative variables while chi-square and Fischer's exact tests (two-tailed) were used for qualitative data, p < 0.05 was considered to be significant. Statistical analyses were performed with Epi-info 6.0 (Center for Disease Control, Atlanta, USA).We aimed to include all residents of the nursing homes. In the study period, 78 subjects were staying in the two nursing homes . However, 7 subjects did not want to participate in the study. Therefore, these seven subjects could not be included.A total of 71 voluntary subjects [54 (76%) male and 17 (24%) female] were included in the study. The mean age was 71. Only 11 (15.4%) of the 71 subjects had a protective antibody level against tetanus. 15 subjects were aged between 60–65 (3 of them had protective antibodies), 22 subjects were aged between 66–75 (2 of them had protective antibodies) and 34 of 71 subjects were aged ≥ 76 years (6 of them had protective antibodies). Protective antibody level decreased with every age group (60–65, 66–75, and ≥ 76) above 60 years old, but there were no statistically significant difference between the five-year age periods above 60 (p > 0.05)  of them had protective antibody levels, only 7 (11%) of 61 subjects who didn't get examined in the emergency room had protective antibody levels (p < 0.05) [Relative Risk = 3.49, 95% Confidence Interval 1.24–9.77].Only 10 (14%) of the 71 subjects had been vaccinated in the last ten years. Five of them had been vaccinated as a result of emergency room visits. Antibody level and demographic properties of subjects were presented in the Table According to the records of the Ministry of Health, 2039 tetanus cases were detected in Turkey between 1980–2002, 462 (23%) of which died . In our In our study group, only 11 15.4%) of 71 subjects had protective antibody levels while the rest were under the risk of tetanus. This resultsuggested that lack of protective immunity among nursing home residents was a significant public health concern. In industrialized countries tetanus has become a rare disease and an infrequent cause of death, mainly due to the implementation of comprehensive immunization programs. But, tetanus is still an important problem for the developing countries, due to poor immunization standards and inadequate hygiene . Deaths .4% of 71Individuals over 60 are usually retired people and spending most of their time in garden or land-work. Tetanus spores may be introduced into the body through a puncture wound, scratch during these works. So they face risk of getting tetanus. Also, 28 % of our subjects still were busy with soil and garden work.According to our results, antibody levels in individuals who had been examined in an emergency room within the last ten years due to injury are significantly higher than who didn't have such a history (p < 0.05). This situation suggests that vaccination programs are not well established in countries like Turkey, thus injuries and accidents expose people over 60 to a great risk of getting the tetanus prophylaxis .Once the existence of tetanus is suspected, intensive, and effective management is essential. The patient should receive intensive care aimed at prevention of muscle spasms, prevention of respiratory tract and metabolic complications, and neutralization of circulating toxins . AssuminThe lack of protective immunity against tetanus among the nursing home residents is a significant public health concern. A vaccination program including every individual over 60 should be charted immediately. After the primary series of 3 doses, protection against tetanus should be sustained by scheduling booster doses routinely in every 10 years . Until tThe author(s) declare that they have no competing interests.All authors read and approved the final manuscript. OK designed the study and drafted the manuscript. AT, KA, analysed the data. OK oversaw the microbiological research. OK and FO interpreted the results of the analysis and critically reviewed the final manuscript.The pre-publication history for this paper can be accessed here:


Creating a new species is a bit like climbing a greased flagpole—it's hard to get started and even harder to keep going. Random genetic variations may introduce slight differences between two groups but, without some means to keep them apart, sexual interbreeding will quickly remix the genes and obliterate the differences. Accidents of geography—the rising of mountains or a course-changing river, for instance—can provide physical isolation, which then enables genetic divergence through the accumulation of mutations, either through natural selection or genetic drift.In contrast, speciation without geographic separation relies on the direct action of natural selection to complete the speciation process by strengthening behavioral differences, a process called reinforcement. One of the most powerful means of completing speciation is through the evolution of mate discrimination.Drosophila. They identify two loci that influence the likelihood that a female will choose to mate with a conspecific male, rather than one of a closely related species.A study by Daniel Ortiz-Barrientos and colleagues focuses on the genetic underpinnings of mate discrimination in Drosophila pseudoobscura and D. persimilis exist together along the west coast of the United States (sympatry), but separately elsewhere . When together, they hybridize and produce sterile males. While D. pseudoobscura males will court females of both species, females prefer conspecific males. This female preference is stronger in sympatric females, an enhancement that presumably evolved by the direct action of natural selection to prevent females from wasting their reproductive efforts producing sterile sons. This variation allowed the authors to conduct a series of genetic crosses among flies of the same species but from different locations. Because the daughters of discriminating D. pseudoobscura females were just as discriminating as their mothers, Ortiz-Barrientos and colleagues concluded that female mating discrimination was inherited as a dominant trait. Further crosses showed that genes responsible for female preference were on the X and fourth chromosomes, and high-resolution mapping refined their locations sufficiently to allow the identification of likely candidate genes. While more work remains to be done, the most promising genes in both regions appear to be involved with olfaction. The fact that one of them, CG13982, is known to up-regulate the other, bru-3, strengthens the case that these are indeed promising candidates.D. pseudoobscura based on the combined response to auditory and olfactory cues. The first of these two layers, weak, or “basal” mating discrimination, has previously been associated with a set of traits for acoustic recognition and mapped to chromosomal regions that are inverted between the two species. Such inversions prevent recombination from purging alleles, thereby contributing to hybrid male sterility. As a consequence, when these species interbreed, they inexorably produce sterile males. The second layer, elucidated in the current study, is “reinforced” mating discrimination, which appears to be related to olfactory cues. This additional system of discrimination helps the first layer to fully eliminate the inevitable cost of producing sterile males. Once the nascent species have started up that slippery pole, reinforced discrimination could provide the traction needed to reach its top.According to their findings, the authors propose a novel model of mating discrimination in


Implantation of the embryo and successful pregnancy are dependent on the differentiation of endometrial stromal cells into decidual cells. Female interleukin-11 receptor α (IL-11Rα) deficient mice are infertile due to disrupted decidualization, suggesting a critical role for IL-11 and its target genes in implantation. The molecular targets of IL-11 in the uterus are unknown, but it is likely that IL-11 signaling modifies the expression of other genes important in decidualization. This study aimed to identify genes regulated by IL-11 during decidualization in mouse uterus, and to examine their expression and localization as an indication of functional significance during early pregnancy.IL11Ra+/+) and IL-11Rα deficient (IL11Ra-/-) littermates by oil injection into the uterine lumen, and gene expression analyzed by NIA 15K cDNA microarray analysis at subsequent time points. Quantitative real-time RT-PCR was used as an alternative mRNA quantitation method and the expression and cellular localization of the protein products was examined by immunohistochemistry.Decidualization was artificially induced in pseudopregnant wild type , biglycan and nidogen-1 (entactin). Immunohistochemistry confirmed increased collagen III and biglycan protein expression in IL11Ra-/- uterus at this time. In both IL11Ra-/- and wild type uterus, collagen III and biglycan were primarily localized to the outer connective tissue and smooth muscle cells of the myometrium, with diffuse staining in the cytoplasm of decidualized stromal cells.Among 15,247 DNA probes, 13 showed increased and 4 decreased expression in These data suggest that IL-11 regulates changes in the uterine extracellular matrix that are necessary for decidualization. Implantation of the embryo, formation of the placenta and successful pregnancy are dependent on the proliferation and differentiation of endometrial stromal cells into decidual cells. Decidualization occurs in response to endometrial and embryonic signals and is thought to involve complex interactions between ovarian steroid hormones, the uterine extracellular matrix (ECM), growth factors and cytokines (reviewed in ). When iIL11Ra-/-) mice are unable to support implantation of either IL11Ra-/- or wild type embryos. The failure of decidualization between days 4.5 and 10.5 in IL11Ra-/- females is characterized by greatly reduced vascular permeability at implantation sites, areas of hemorrhage, impaired secondary decidual zone formation, absence of mesometrial decidualization and aberrant infiltration of trophoblast giant cells [IL11Ra-/- mice show some mesometrial decidualization. Females homozygous for a hypomorphic IL-11Rα allele also show reduced decidualization, with decreased cell proliferation, progressive degeneration of the deciduae, infiltration of trophoblast giant cells and absence of placental formation [Female mice with a null mutation in the gene encoding interleukin-11 receptor α (IL-11Rα) are infertile due to disrupted decidualization , suggestnt cells . Althougormation . Neitherormation ,9.Interleukin-11 is a multifunctional cytokine, initially described as a bone marrow stroma-derived hematopoietic growth factor . IL-11 sLocalization and expression of IL-11, IL-11Rα and gp130 in human endometrium across the menstrual cycle suggests a role for this cytokine in decidual transformation in preparation for pregnancy -20. LeveThe molecular targets of IL-11 in the uterus are unknown, but it is likely that IL-11 signaling modifies the expression of other genes important in decidualization. This study aimed to identify genes regulated by IL-11 during decidualization by cDNA microarray, and to examine their expression and localization by immunohistochemistry, as an indication of functional significance during early pregnancy.IL11Ra-/-) had been previously generated by gene targeting, and serially crossed more than 10 generations onto a C57BL/6 background [IL11Ra+/+) and IL-11Rα deficient (IL11Ra-/-) mice, which were identified by Southern blot analysis of genomic DNA obtained from tail biopsies [Mice deficient in IL-11 receptor α  and 80 mg/kg ketamine hydrochloride  in sterile phosphate-buffered saline (PBS). Anesthesia was reversed with 250 μg yohimbine hydrochloride and 400 μg 4-amino pyridine  in sterile PBS.IL11Ra+/+ and IL11Ra-/- littermates at 8–12 weeks of age (n = 4 per genotype per time point) were mated with wild type vasectomized males to induce pseudopregnancy, with the day of plug detection designated day 0. At approximately 1400 h on day 3, decidualization was induced throughout both uterine horns of each animal by injection of 20 μl of sesame oil  into the lumen of each uterine horn via a 26-gauge needle inserted just distal to the utero-tubal junction. Mice were necropsied prior to surgery (0 h) or at 18 h, 24 h or 48 h following artificial decidualization. These time points prior to the onset of secondary decidualization were chosen to ensure similar cellular composition of the IL11Ra+/+ and IL11Ra-/- uteri. Whole uteri were cleaned of fat and weighed. A section of each uterus was either fixed in 10% phosphate-buffered formalin overnight or Carnoy's fixative for 2 h and processed to paraffin wax, or snap frozen in liquid nitrogen for subsequent RNA isolation.Female IL11Ra+/+ (n = 4/time point) and IL11Ra-/- (n = 4/time point) mice were statistically analyzed using GB-Stat 6.5 (Dynamic Microsystems, Inc., Silver Spring, MD). Following Bartlett's test for homogeneity of variance, uterine weight was used as the dependent variable and genotype and time as the two independent variables in a two factor analysis of variance. Bonferroni multiple comparison testing was used to compare uterine weight across time in each genotype and between genotypes at each time point. A two-tailed p value of less than 0.05 was considered a significant difference.The wet weights of whole uterus from 260:OD280 = 1.8–2.0). Each artificially decidualized IL11Ra+/+ or IL11Ra-/- uterus was processed individually for microarray hybridization. A reference pool of RNA was prepared from wild type unstimulated uteri (n = 16). The experimental design used indirect comparisons between IL-11Ra+/+ or IL11Ra-/- and the reference pool.Total RNA was extracted from whole uterus by acid guanidinium thiocyanate-phenol-chloroform extraction , incorpoIL11Ra+/+ or IL11Ra-/- cDNA (Cy3) and reference cDNA (Cy5) were competitively hybridized to the microarrays in the presence of 1 mg/ml Cot1 DNA , 10 mg/ml salmon sperm DNA (Gibco-BRL) and 10 mg/ml polyadenylic acid (Sigma). Hybridizations were repeated using the alternate dye combinations to account for differential fluorescent dye incorporation. After washing, slides were scanned using an Axon GenePix 4000B microarray reader (Axon Instruments, Union City, CA) and GenePix Pro 4.0 software (Axon Instruments) to generate pairs of 16-bit tagged image file format (TIFF) files. Following manual quality control for hybridization artefacts, red (Cy5) and green (Cy3) mean foreground and median background fluorescence intensity measurements for each spotted DNA sequence were extracted for export to the statistical programming environment R 1.5.1.Total RNA (10 μg) served as a template for the synthesis of aminoallyl-cDNA, which was then coupled to a fluorescent dye ester (Cy3 or Cy5) as described by the manufacturers of the CyScribe cDNA Post Labelling Kit (Amersham Biosciences, Buckinghamshire, England). Microcon-30 size exclusion columns (Millipore, Billerica, MA) were used to initially concentrate RNA samples, and to purify the cDNA probes prior to and following the dye-coupling reaction. Slides were printed with sequence-verified duplicate spots of the NIA 15K cDNA clone set  at the AIL11Ra+/+ (WT) and IL11Ra-/- (KO) uterus at time points following artificial decidualization was determined using the normalization and analysis functions of the statistical language R [R and G) from 4 slides were read into R using the read.genepix and init.data commands. The stat.ma function was used to calculate the log-ratios of expression (M = log2R - log2G) and average log-intensity (A = (log2R + log2G)/2) for each spot, and to normalize the red and green channels relative to one another using print-tip loess normalization [MA plots of each slide were used to determine the effectiveness of this normalization method in adjusting for sources of variation arising from dye bias and print-tip effects.Differential gene expression between nguage R  and the nguage R . For eaclization . Diagnost-statistic and B-statistic for each gene, to give a log odds ratio of difference in mRNA expression between IL11Ra+/+ and IL11Ra-/-. Genes with a log odds score of greater than 3  in both replicates were considered to be significantly up- or down-regulated in IL11Ra-/- uterus compared to wild type.Given the indirect dye swap design  of theseIL11Ra+/+ (n = 2) and IL11Ra-/- (n = 2) uterus at 48 h of decidualization and used for microarray analysis, and an additional two samples per genotype prepared in the same way were used for validation of the microarray data by real-time reverse transcription polymerase chain reaction (RT-PCR). All samples (n = 4/genotype) were further purified through an RNeasy Spin Column , and quantitated by RiboGreen Assay (Molecular Probes, Eugene, OR) according to the manufacturer's instructions, prior to triplicate reverse transcription reactions.Total RNA samples extracted from Total RNA (1 μg) was reverse transcribed at 46 C for 1.5 h in 20 μl reaction mixture using 100 ng random hexanucleotide primers and 6 IU AMV reverse transcriptase  in the presence of cDNA synthesis buffer (Roche), 1 mM dNTPs (Roche), 10 mM dithiothreitol (Roche) and 10 IU ribonuclease inhibitor . The resulting cDNA mixtures were heated at 95 C for 5 min before storage at -20 C in small volumes to avoid freeze-thawing. Negative controls were performed by omission of reverse transcriptase.IL11Ra+/+ and IL11Ra-/- uterus at 48 h of decidualization, PCR was carried out using a Roche LightCycler. Prior to LightCycler analysis, standard cDNA for each gene of interest was generated using a PCR Express block cycler (Thermo Hybaid Instruments, Franklin, MA). A 1 μl aliquot of RT product was amplified in a total volume of 40 μl using 4 μl of 10 × PCR Reaction Buffer , 62.5 μM dNTPs (Gibco-BRL, Life Technologies), 10 pmol sense and antisense primers  was added to sterile capillaries to a total volume of 20 μl containing SYBR Green I, dNTPs, Taq DNA polymerase and reaction buffer , supplemented with 5 pmol of specific sense and antisense primers Table  and an oIL11Ra+/+ and IL11Ra-/- uterus. Following normalization, levels of RNA for COL3A1, BGN, SPARC and NID1 in IL11Ra-/- compared to wild type were statistically analysed using the paired t-test function of GraphPad Prism 3.0 (GraphPad Software, San Diego, CA). A two-tailed p value of less than 0.05 was considered a significant difference.Triplicate RT reactions for each sample, the standard curve and a no RT negative control were analyzed in the same run, and each run was repeated once. A mean value for the initial target concentration of each sample was calculated using the fit points function of the LightCycler software. The mean concentration of 18S rRNA for each sample was used to control for RNA input, as it is considered a stable housekeeping gene and was not altered in expression between IL11Ra+/+ and IL11Ra-/- uterus collected at 48 h of decidualization, using specific antibodies (n = 5 mice per genotype, n = 3 sections per mouse). The morphology and cellular localization of artificial decidualization in IL11Ra+/+ and IL11Ra-/- uterus was also determined by immunostaining for the decidual marker desmin [To confirm differential expression of collagen III, biglycan, SPARC and nidogen-1 at the protein level, immunohistochemistry was carried out on transverse sections of r desmin . Primaryr desmin , gift frFive micron sections of Carnoy's-fixed  or formalin-fixed (for SPARC, nidogen-1 and desmin) uterus were mounted on poly-L-lysine-coated glass slides, deparaffinized and rehydrated through a series of graded ethanols. In an humidified chamber at 25 C, sections were incubated for 10 min in 3% hydrogen peroxide to block endogenous peroxidase activity, then for 30 min (SPARC and desmin) or 1 h (collagen III, biglycan and nidogen-1) in 10% serum of the species in which the secondary antibody was raised  and 2% mouse serum in one of the above buffers. Sections were then incubated with primary antibody or negative at 4 C overnight (collagen III, biglycan, nidogen-1 and SPARC) or 30 min at 25 C (desmin), then washed in TBS/Tween prior to secondary antibody incubation for 30 min (SPARC and desmin) or 1 h (collagen III, biglycan and nidogen-1) at 25 C. Sections were again washed in TBS/Tween, then the secondary antibody detected using the Vectastain ABC Elite/HRP Kit  or the StreptABComplex/HRP Kit  according to the manufacturer's instructions. Protein localization was visualized using the Liquid DAB-Plus Substrate Chromogen System (DAKO), with Harris hematoxylin (Sigma) counterstain.p < 0.01) at the final time point  in uterine weight at 48 h of artificial decidualization between IL11Ra+/+ (96.9 +/- 9.8 mg) and IL11Ra-/- (30.4 +/- 7.7 mg).Following the injection of oil into the wild type pseudopregnant uterus, a progressive increase in uterine weight was observed from 0 through to 48 h, reaching statistical significance p < 0.0 at the fIL11Ra+/+ and IL11Ra-/- uterus artificially decidualized for 0, 18, 24 or 48 h (n = 2/genotype/time point) was used as a template for the hybridization of NIA 15K cDNA microarrays. Figure p = 0.05). At 0 h, prior to application of the decidualizing stimulus on day 3 of pseudopregnancy, there were no reproducible differentially expressed genes between IL11Ra+/+ and IL11Ra-/- , COL3A1  and IL11Ra-/- (1.66 +/- 0.09 fg/μl) uterus approached statistical significance at p = 0.0708.To confirm the altered mRNA expression of the ECM genes COL3A1, SPARC, BGN and NID1 at 48 h of decidualization, quantitative real-time RT-PCR was carried out using the same RNA samples used in the microarray analysis, plus two additional RNA samples of each genotype, collected in the same way. At a significance level of 3A1 Fig. , SPARC . GrowthIL11Ra-/- compared to wild type uterus, this effect was not statistically significant when real-time RT-PCR was used as an alternative quantitation method. Many factors may contribute to discrepancies between cDNA microarray and real-time RT-PCR data. There are major differences in the approach to mRNA quantitation used by the two techniques. Using cDNA microarray, the mean fluorescence intensity ratio for each gene in IL11Ra-/- or IL11Ra+/+ uterus was calculated relative to a reference pool, and the ratio of IL11Ra-/- to IL11Ra+/+ determined by the use of computational algorithms. When quantitating the same mRNA species by real-time RT-PCR, a standard curve of known concentration was used to infer the absolute abundances of mRNA in the IL11Ra-/- and IL11Ra+/+ samples, which were then normalized for RNA input.While the microarray data showed consistent, reproducible upregulation of COL3A1, BGN, SPARC and NID1 in Real-time RT-PCR was chosen for cDNA microarray validation in this study because it has higher sensitivity and lower RNA requirements than Northern blot, but the lack of agreement between the two methods is not unusual. It is well recognized that fold change values for a given gene may vary widely, even between two different microarray techniques -43. In uThere are a number of compelling arguments both for and against conducting corroborative studies for microarray data, and there is good evidence that the data is highly reliable when the experimental design and statistical analysis is sound . In asseIL11Ra-/- uterus is supported by existing data in the literature. Collagen III is a fibrillar collagen with known roles in differentiation and migration [For each of the genes examined, upregulation during defective decidualization in igration , and togigration . Consistigration , and secigration . The redigration  may faciigration . This deigration . In the igration . This isIL11Ra-/- mice could contribute to impaired decidualization.During the human menstrual cycle, collagen III immunostaining is higher in the proliferative compared to the secretory phase , indicatIL11Ra-/- uterus at 48 h of decidualization, it appears that successful decidualization involves downregulation of COL3A1 transcription.Using microarray analysis, the mRNA encoding procollagen III α1 (COL3A1) has been previously shown to decrease in abundance in the mouse uterus at estrus , and betIL11Ra-/- mouse uterus was associated with the upregulation of biglycan mRNA, the activity of this proteoglycan in the ECM may inhibit the decidual response.Biglycan is a small leucine-rich proteoglycan, which binds to type I  and typeDecidual cells are known to express nidogen-1 as part of the pericellular basement-membrane laid down during decidualization . The maiSPARC (osteonectin/BM-40) is described as a matricellular glycoprotein, in that it binds to both cells and ECM to regulate cell-matrix interactions . Like otUsing mRNA and protein expression studies alone, it is not possible to determine whether IL-11 affects ECM molecule expression directly or indirectly. The presence of STAT binding sites in the promoters for COL3A1, BGN, NID1 and SPARC would confirm that a direct interaction is possible, but would not establish that the interaction is occurring in the uterus during decidualization. Given that IL-11 is a secreted cytokine with autocrine and paracrine activity, a direct effect on ECM molecule transcription would be dependent on coincident expression patterns of the ECM molecules, IL-11 and its receptors. Maximal expression of IL-11 and IL-11Rα mRNA has been reported in the predecidual and decidual cells of the mouse implantation site by in situ hybridization . ExpressIL11Ra-/- uterus compared to wild type. Previous in vitro studies have indicated that IL-11 inhibits MMP-1 and -3 protein in human synovium [Indirect effects of IL-11 on ECM composition could be mediated by matrix metalloproteinases (MMPs) and/or their inhibitors. Despite their presence on the NIA 15K microarray, differential expression was not observed for MMP-2, MMP-9, TIMP-2, or TIMP-3 in the synovium , and enhsynovium . IL-11 dsynovium , or indusynovium , but tissynovium . While nThis investigation of the downstream targets of IL-11 during mouse decidualization has uncovered previously unknown interactions between IL-11 and uterine ECM composition. Dysregulation of collagen III, biglycan, nidogen-1 and SPARC in the absence of IL-11 signaling at the time of decidualization may indicate essential functions for these molecules during the implantation process in mice. Functional studies using mouse and human endometrium may further clarify the mechanisms of IL-11 action on the ECM during this critical time in embryo implantation. By elucidating the role of IL-11 regulated genes in decidualization, future work may identify potential new targets for the manipulation of human fertility.IL11Ra+/- breeding pairs. LAS conceived of the study, participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.CAW carried out all the experimental work and statistical analysis and drafted the manuscript. LR provided the heterozygote


Medical outliers present a medical, psychological, social, and economic challenge to the physicians who care for them. The determinism of Stoic thought is explored as an intellectual basis for the pursuit of a correct mental attitude that will provide aid and comfort to physicians who care for medical outliers, thus fostering continued physician engagement in their care.The Stoic topics of good, the preferable, the morally indifferent, living consistently, and appropriate actions are reviewed. Furthermore, Zeno's cardinal virtues of Justice, Temperance, Bravery, and Wisdom are addressed, as are the Stoic passions of fear, lust, mental pain, and mental pleasure. These concepts must be understood by physicians if they are to comprehend and accept the Stoic view as it relates to having the proper attitude when caring for those with long-term and/or costly illnesses.Practicing physicians, especially those that are hospital based, and most assuredly those practicing critical care medicine, will be emotionally challenged by the medical outlier. A Stoic approach to such a social and psychological burden may be of benefit. Medical outliers are defined in health care reimbursement, especially in the prospective payment system, as those patients who require an unusually long hospital stay or whose stay generates unusually high costs, i.e., the most severely ill . AccordiFinancial concerns have overwhelmed the American medical educational system and hospitals ,5. MedicEric Cassell has pointed out that medical practice is now being shaped by commercial considerations; the issue of financing health care dominates all facets of medicine, including education, research, the relationships between physicians, and the relationships between physicians and their patients . NonetheIn a previous debate an argument was made, using the works of Kant and Hegel, that physicians have an obligation to treat medical outliers . Here anStoics will argue that becoming a physician is predetermined by Nature, or God, and, that having occurred, it must also be predetermined that a physician will care for medical outliers. Thus, the only thing physicians can alter, or control, is their attitude. Zeno, the founder of Stoicism, explains,"A man's excellence or virtue does not depend on his success in obtaining anything in the external world; it depends entirely on having the right mental attitude toward things" .logos, the ability of humans to think, plan, and express themselves, was inherent in the cosmos. Therefore, logos is part of Nature, or God,Stoicism teaches that the universe is rational, that it can be explained rationally, and organized rationally. Stoics taught that "God and man are related to each other at the heart of their being rational agents. If a man fully recognizes the implications of this relationship, he will act in a manner which wholly accords with human rationality at its best, the excellence of which is guaranteed by its willing agreement with nature. This is what it is to be wise...." .Stoics believed that a cosmic thread related every event to another and that such a premise allows a human to live a life at one with Nature, or God. In acquiring such an understanding a human could strike an accord between his or her attitudes, actions, and the course of events.The reason for having a good mental attitude and the understanding that a cosmic thread relates all actions and events, according to the Stoics, is that God rules all and nothing can occur unless God wills it. Accepting everything that happens to one's self will bring contentment,"So the good man will accept everything, knowing that it is not only unalterable, since Fate determines all, but also the work of God, the perfect being; namely that his happiness depends entirely upon himself, and not at the mercy of other persons or the play of outside forces. What brings happiness is to have the right attitude, to choose the right actions, to aim correctly at the mark" .Therefore, to "try" is within a man's power, and good intentions are indeed enough according to Stoic philosophy. The ability to succeed is not necessarily within man's grasp, but his attitude is his own doing. If a person does his or her best and has no self-reproach for this effort, then he or she is one with Nature, or God.There are difficulties with accepting Stoic philosophy, which will be reviewed later. Nonetheless, the Stoic view of a physician's approach to the care of the medical outlier will be advocated as one that may allow a physician to be successful in outlier care. Success does not refer to monetary remuneration, but an approach that will allow a physician mental comfort (a good attitude) in such an advocacy.Physicians must understand the concepts of good, the preferable, the morally indifferent, living consistently, appropriate actions, the cardinal virtues, and the passions to successfully apply the Stoic view.What is good? Any physician pondering this question may come up with answers such as a lucrative medical practice, minimal on-call days, or passing medical board examinations. These things are good, especially when compared to poverty, sickness, or unemployment. However, the Stoics believed what was "good", was also morally perfect (virtue, virtuous acts and virtuous people). Virtue and virtuous things belonged in a league of their own. If you were virtuous, according to the Stoics, you were "good", therefore happy, and this was moral perfection. If you were virtuous you always did what was morally right. Things that are "bad" are morally imperfect (not virtuous); here we speak of evil and wickedness, not poverty, illness, or death.The Stoics' view of good and bad were extremes of perfection and imperfection. Beauty, wealth, a good job, and a good marriage were things that were preferable, but not morally "good". Illness, poverty, and death were less preferable, but not morally "bad". However, neither the preferable, nor the less preferable were considered "good" or "evil"; they were considered morally indifferent,Right action  is a matter of choice concerned with morally indifferent things – will you look for wealth or accept poverty, marry or remain a bachelor, live or die? – and choice between absolutely different alternatives would not involve knowledge or reason.... Virtue then can consist in the effort to obtain these things that have value and avoid their contraries, and knowledge can be knowledge of what is to be preferred. But since things of this sort are not "good" or "bad", it is of no importance whether one has them or does not have them, as far as goodness is concerned. The good intention is enough; achievement may be impeded by forces outside a man's control" ["Goodness, however, and knowledge, although they had value of a unique kind, could not be the only things to have value. control" .It can generally be agreed upon that caring for medical outliers, ideally, is a "right action" because we are dealing with sickness and economics, i.e., morally indifferent things that are not preferable. Many times caring for medical outliers is something we must do to keep our position, fulfill our contract, or to avoid a lawsuit. However, if we speak of doing what is right in the virtuous, or morally perfect sense, caring for medical outliers must be more than moral indifference. It must be an act of virtue, but can only be so if the physician is doing it out of the deepest sense of duty. Virtue in this case means having the right mental attitude toward outlier care and understanding that it is more than an ordinary good, it is an action that stands morally in a class of its own.Philosophers in ancient Greece would inquire, "What is the goal of a perfect life" . The StoZeno explained that "the single plan by which life should be lived must be a plan formed by correct reason, and this would be one that is natural in the sense that it accords both with man's nature and with universal nature" . In advoTo be happy or content in the care of medical outliers a physician must have this consistency of life and an absence of conflict in this regard. To be able to understand this concept and accept it, the physician must be aware that he or she is a part of the whole. For example, let us say a flower flourishes in a garden; we understand what "to flourish" means. Also, let us say we observe a group of birds, some are healthy and some are not. Therefore, we know what the natural condition, or norm, for a flower and a bird should be. This norm is good. So each thing, flower, or bird has a good that is a universal. There is a universal condition or nature that is appropriate for each thing,"Eating hay is natural to horses, but not to men. It accords with universal nature that horses should eat hay and that men should speak a language. But the former is inappropriate to men and the latter to horses. Universal Nature sanctions a norm for particular things – the nature of plants, animals and men – by reference to which they can be said to attain or not to attain their individual ends" .Thus we can understand what is meant by "the part". Man is born and man will die. Man fights the Universal Nature of death, a particular "norm" of humans. In struggling against his role, or his "part" in nature, man may do extraordinary things to keep himself healthy or alive. A man may decide to preserve his home or his child's education by not having (paying for) health insurance. A physician, for example, may decide that a patient needs an organ transplant. The physician will engage this struggle whether or not the patient has health insurance, and the end result may make the patient a medical outlier, physiologically and/or economically. Universal Nature may sanction a norm for particular things, but humans, and especially physicians, often struggle against their role as a "part" and come into conflict with nature.Each flower, bird, person, or physician is a part of Nature's whole. Contrary events may happen to an individual "part". Birds are hunted and eaten, flowers are cut and put into a vase, and severely ill humans may end up on long-term ventilatory support. The Stoics believed that such events are part of nature's order. Such a view may be contrary to a human observing the whole of nature. However, if the human perspective is removed from an event then, "From the perspective of the whole even such conditions are not unnatural, because all natural events contribute to the universal well-being" .The Stoics combine their views of the part and the whole. They view the whole as perfect and the nature of perfect requires inequities and incompatibilities; nothing that happens to a human is disadvantageous to him or her, nor is it a disadvantage to nature. Nature is perfect, so according to Stoicism, suffering does not occur for its own sake, but "it is necessary to the economy of the whole" .For physicians to live consistently they must understand their place in nature. Stoics would explain to today's physician that their contentment in dealing with medical outliers depends on this understanding of nature. If this is understood and accepted there will be no conflict and "living consistently" will be attainable.appropriately is not in itself either good or bad in the sense of being morally "good" or "bad" [Most physicians hope to make the right, or moral, decisions in regard to their actions. Caring for medical outliers is a choice that has to be made by many physicians. Stoics believed it was always appropriate to act virtuously, but acting virtuously can only occur when a man is "perfectly good", and since men are not "perfectly good" then physicians cannot act virtuously, in other words, be morally perfect. However, Stoicism has made a niche for appropriate actions, "to act virtuously is always morally good, and to act faultily is always bad, to act or "bad" . Even thA physician may care for a medical outlier, but it is not necessarily a morally good action if he or she does it without the complete understanding of why it is the right thing to do. In other words, taking care of a medical outlier is a just action, and thus an appropriately good action, but only if the physician is doing it without duress or not being mandated to attend to the patient .Irrational forces plague a man's mind according to Plato, and these forces have to be controlled before a man could have the needed knowledge to act virtuously. The Stoics, however, did not feel this was true. They felt that if a man could be trained to think correctly, then he could learn to act virtuously.Justice was wisdom concerned with distribution. Temperance (self-control) was wisdom concerned with acquisition. Bravery was wisdom concerned with endurance. Wisdom was defined as "knowledge of what should and should not be done, or knowledge of what is good or bad or neither" [Zeno went on to define four cardinal virtues that were necessary for a man to acquire to be successfully trained to think correctly so that he could act virtuously. He used Plato's work as a basis and he defined the four virtues in terms of the fourth virtue, wisdom. neither" .Medical outliers require justice. Physicians must be able to distribute their actions  fairly and equitably to all those who are in need of such services. Temperance, or self-control, must be learned or acquired. Physicians have to control their emotions when assigned to a medical outlier, having their patients turn into medical outliers, or seeing other parties' responses to medical outliers. Physicians should allow neither anger, frustration, anxiety, nor fear to overtake them. Enduring the endless days, weeks, or months of caring for a medical outlier certainly requires bravery and stamina. Fielding the unending phone calls and the constant re-tuning of a patient's hemodynamic status can be of marathon proportions. Wisdom is what should or should not be done and what is good or bad must not only apply to the type and amount of medical care, but it should also encompass the virtue of the acts of justice, compassion, and care given to the medical outlier.pathos) or what many of them called a mental disturbance. The Stoic "passion" is defined as an excessive uncontrollable drive due to an overestimation of the worth of the "indifferent" things (or events) mentioned previously. Nonetheless Stoics taught that to have great affection was indeed desirable, but at the same time one should remain passionless. Animals are driven to an action because of a stimulus, but in man such a stimulus (or impulse) requires the mind to accede to the stimulus. The Stoics found this to be important because they felt it to be a point of distinction between humans and animals. To the Stoics all living animals were compelled to respond to stimuli by their psyche, a mix of fire and air that was responsible for the functions of living animals . There are times, however, when a man's mind is out of control and his passions become excessive [Physicians need emotion. The Stoics did not disagree, but they wished to eliminate passion . The passions were explained by F.H. Sanbach,"Fear is a contraction of the psyche caused by the belief that something bad is impending. It causes paleness, shivering, and thumping of the heart. But the belief is false: what is feared is not what a Stoic calls "bad", but one of the morally indifferent things, e.g. death, pain, ill-repute. Fear is the result of exaggerating their importance, of believing they will bring real harm, whereas they do not affect man's essential moral being and if they come are to be accepted as part of the great plan of nature. Lust is a longing for something believed to be good, but again is falsely so believed, since the supposed good is morally of the psyche. Mental pain is a contraction of the psyche resulting from the belief, again erroneous, that something bad is present.... Pleasure was defined as an irrational expansion of the psyche caused by the supposed presence of something good.... What is thought to be good is not in fact good, but at the best acceptable" .In many instances the passions do come into play when a physician cares for a medical outlier. Fear affects the physician from several perspectives. Physicians fear for adverse outlier outcomes, not only for the patient's sake and that of the family, but also out of concern for potential litigation, non-reimbursement for services rendered, and long hours incurred in the care of the patient.In regard to lust (desire), something believed to be good, but falsely so, several points can be made. The Stoics spoke of many types, or species, of lust, anger being foremost among them. This species of lust is very appropriate to discuss in regard to care of the medical outlier. Physicians do get angry and occasionally act out when challenged. Outliers involve a large investment of emotion, time, and a potential loss of income on the part of the physician . Also, hostility toward staff for small deviations from the plan of care may occur more frequently than the staff would like. As the patient's course of illness drags on physicians may have anger for the patient and the family . Such anger occurs because the patient does not improve or improves too slowly. There may also be anger towards the family for asking too many questions or questioning the plan of care the physician is following. In addition, the family may also want other physicians consulted or more time from their current physician. What is, in fact, happening is that the family is merely trying to exert control over what little they can yet control.Mental pain or anguish is self evident in medical outlier care. The previous passions of fear and lust (the species of anger) contribute to the burden of mental pain. The species of mental pain that Stoics address include grief and pity, two potentially powerful distorters of judgment for physicians.Mental pleasure in caring for those that are seriously and/or chronically ill is not as self-evident. Again we do not speak here of physical pleasure. The species of mental pleasure include "pleasure at unexpected 'benefits', pleasure at other people's misfortunes, pleasures caused by deceit and magic" . PhysiciThere is no doubt that medical outlier care evokes passions. There is little for a physician to do but his or her best in regard to giving health care. Much is out of the physician's control, and therefore much of Stoic thought is applicable; control the passions, keep a good attitude, have an open mind about plans of care, have an open mind as to who can participate in decisions , understand what is a reasonable outcome, and remember that is does not matter who gets credit for good outcomes .If all human events and actions are predetermined how are human freedoms and free will to be addressed? Universal causation is the bedrock of Stoic philosophy. If human attitudes and beliefs are within an individual's power or sphere of influence, is this truly congruent with Stoic determinism?Robert L. Arrington illustrates the human attitude towards sickness as a foible in Stoic thought . Illnessit is not possible for us to perceive it as a matter of indifference. If, on the contrary, the causes lead us to assume the attitude of indifference, then it becomes impossible for us to see the illness as misfortune. Either one of the sets of courses or the other must exist, from which it follows that it is either impossible for us to feel misfortune or impossible for us to feel indifference. If one of these options is impossible, the attitude we take is necessary in which case we really didn't have any options at all. And without options or choices, there is no thing as freedom or voluntary behavior. And, so it seems, our attitudes and beliefs are not in our power" ["And if the causes that exist prior to our forming the attitude lead us to perceive the illness as misfortune, r power" .This argument regarding whether universal causation and determinism is consistent with a free will has been debated for over 20 centuries. Today there are philosophers on both sides of the issue.Another flaw is the Stoic approach to evil. Stoics simply tell us it does not exist; events may seem evil, but they are not. Stoics teach that only the human perspective allows the interpretation that evil exists. Religions of the world, many philosophers, and people who have viewed and/or endured suffering cannot agree with the Stoics.A further distortion in Stoic thought involves the idea that the life of virtue is the only "good" life. What about the "preferred" things that we as humans know make our lives better? What is wrong with "attaining the goals of impulse" ? There wAnd, finally, the Stoics felt the universe was rational and in unity. A divine thread ran through the cosmos connecting everything and everybody. Many philosophers cannot accept this concept. However, as we see the progression of this line of reasoning as it regards the study of the "string" theory in physics and the further work and modification of Einstein's views of relativity, we realize that there may be a mathematical basis to existence. The Stoics may be criticized about their "thread" through the cosmos, but when we discuss how time "bends" and describe gravity as "curved space" the critics of Stoicism may be tightrope-walking this same thread.The health care rendered to medical outliers can be for a substantial length of time and cost to the medical practitioner. The intensive interaction with the patient, his or her family, consultants, nurses, other ancillary staff, and the institution can sap the performance of involved physicians.Placing the argument for the predetermination of events aside, if one is a physician that is involved in a hospital-based specialty, critical care medicine, or as surgeon, the fact of the matter is that medical outliers will come to your door. By fate or by choice physicians in the above-mentioned areas will be engaged with outliers. As the Stoics point out, the future is coming at you and there is nothing you can do about it, except adjust your attitude.While engaged in such an endeavor physicians will "try" to do their best, hopefully without self-reproach as to their efforts. Physicians, as humans, cannot be "good", or morally perfect. They become tired, hungry, worry about the bills, their children, their practice, hospital policies, etc. Nonetheless, most physicians realize and understand what appropriate actions are necessary in regard to the most ill and poorest of patients. Also, though physicians may not be morally perfect (virtuous in the Stoic sense), they know what is "preferable" for their patients, i.e., to get well, go home and be with their families. These are morally "indifferent" things, but as Stoics point out, "virtue then can consist in the effort to obtain these things that have value and avoid their contraries, and knowledge can be knowledge of what is to be preferred" .As mentioned previously, physicians frequently struggle against their role as a "part". By the mere fact that physicians acknowledge struggles with or against insurers, patients, families, colleagues, ancillary staff, and institutions there is realization that they are part of a "whole", but at the same to "live consistently" requires an understanding of one's role in nature and the need for absence of conflict. In the struggle to help the critically ill, the chronically ill, or the incredibly poor, avoiding conflict is challenging.The cardinal virtues of justice, temperance, bravery, and wisdom come with upbringing, education, and life experiences. Lacking the proper intellectual or nurturing environment will not allow the flourishing of these virtues in an individual. In schools of medicine the faculty attempt to inculcate these virtues in the students, although they are not always successful.The Accreditation Council for Graduate Medical Education (ACGME) has recently mandated six core competencies for resident education . One of Justice is a far-reaching concept that more physicians need to embrace. It is difficult to teach and is best acquired through experience of its antithesis. Temperance, or self-control, can be mandated by medical staff guidelines and licensing boards, but this virtue actually needs to have been ingrained before medical school. Bravery is something that residency training seeds in a physician through the frequent facing of dying or hostile patients that come through a medical practice, especially at an academic institution that cares for the disenfranchised. Wisdom will come with time and maturity. Both are needed to acquire wisdom. Physicians, medical students, and residents can, indeed, be taught to think correctly, as the Stoics emphasized. However, individuals will have varying degrees of success depending their rearing/nurturing and educational environment.Emotion is necessary to physicians, but the passions, those uncontrollable mental disturbances due to overestimation of the value of "indifferent" things, may blind them in their judgment and decision-making. It is important that when making important decisions in regard to medical outliers that the passions be "checked".To avoid frustration, disappointment and unhappiness in the practice of medicine as it regards medical outliers, physicians must do two things: (1) control things that are within their power (attitudes, desires, beliefs), and (2) be indifferent to the things that they cannot control  .Even though Stoicism has evoked controversy for over twenty centuries it is relevant to a physician who must juggle patients, procedures, therapies, and colleagues in the care of a patient who has maximally taxed medical insurers, institutions, other practitioners, and their own families.Insurers and institutions may have financial burdens, but those providing patient care, especially physicians, bear a disproportionate slice of the mental anguish associated with the care of medical outliers. Here an argument has been made that applying the philosophical tenets of Stoicism to the physician's intellectual pursuit of how to deal mentally with the care of medical outliers is appropriate. Physicians that are hospital based and those practicing critical care medicine may well be the providers most emotionally challenged by outliers. A Stoic approach to such a social and psychological burden may be helpful.The author(s) declare that they have no competing interests.TJP is responsible for the manuscript in its entirety.The pre-publication history for this paper can be accessed here:


H. pylori infection using only non-invasive tests. We compared the diagnostic value of non-invasive and endoscopy based H. pylori tests in a late post-treatment setting.Guidelines for management of peptic ulcer patients after the treatment are largely directed to detection of 13C-UBT tests were performed.Altogether 34 patients with dyspeptic complaints were referred for gastroscopy 5 years after the treatment of peptic ulcer using a one-week triple therapy scheme. The endoscopic and histologic findings were evaluated according to the Sydney classification. Bacteriological, PCR and cytological investigations and H. pylori positive by 13C-UBT test, PCR and histological examination. On endoscopy, peptic ulcer persisted in 4 H. pylori positive cases. Among the 6 cases with erosions of the gastric mucosa, only two patients were H. pylori positive. Mucosal atrophy and intestinal metaplasia were revealed both in the H. pylori positive and H. pylori negative cases. Bacteriological examination revealed three clarithromycin resistant H. pylori strains. Cytology failed to prove validity for diagnosing H. pylori in a post-treatment setting.Seventeen patients were defined H. pylori colonisation, PCR and bacteriology as they have shown good concordance with 13C-UBT. Moreover, endoscopy and histological investigation of a gastric biopsy have proved to be the methods with an additional diagnostic value, providing the physician with information about inflammatory, atrophic and metaplastic lesions of the stomach in dyspeptic H. pylori positive and negative patients. Bacteriological methods are suggested for detecting the putative antimicrobial resistance of H. pylori, aimed at successful eradication of infection in persistent peptic ulcer cases.In a late post-treatment setting, patients with dyspepsia should not be monitored only by non-invasive investigation methods; it is also justified to use the classical histological evaluation of  Endoscopic biopsies alone are not considered adequate for confirming eradication of bacteria, although they might provide additional information about gastritis and dysplasia [H. pylori status [Treatment of peptic ulcer in accordance with relevant guidelines is becoming a common task for general practitioners -6. In a 13C-UBT) -5. In a er cases . Moreoveysplasia . Use of i status .13C-UBT) and endoscopy-based diagnostic methods  for monitoring patients after eradication therapy in a late post-treatment setting.Our aim was to assess the diagnostic value of different non-invasive (13C-UBT, endoscopy, biopsy, bacteriology, PCR and cytology). The studied patients were not NSAID users.The study group was formed of 134 consecutive peptic ulcer outpatients who had been treated by 7-day triple therapy with metronidazole, amoxicillin and omeprazole in 1996. The group was observed at the outpatient department of Tartu University Hospital at 4 weeks, at 1 year (1997) and at 5 (2001) years after treatment . Five yeThe patients passed the Gastrointestinal Symptoms Rating Scale (GSRS) test  in a val13C-UBT drinking 100 mg 13C-urea; the test meal was citric acid and the time of specimen collection was 30 min. The test was provided, according to a standard protocol, from the Helsinki Keskuskatu Laboratory, Finland. The ratio of 13CO2 to 12CO2 in expired breath was measured by mass spectrometry and expressed in ml/mmol/kg (δ). An automated breath 13C analyser (ABCA) with chromatographic purification and a single inlet isotope ratio mass spectrometer (IRMS) were used. A difference of 5‰ in the content (δ13C) was considered positive for H. pylori infection.The subjects passed The procedure was performed with the gastroscope Olympus-GIF 21. All mucosal defects were registered according to the Sydney classification for endoscopic evaluation . GastricH. pylori colonisation were evaluated on a three-grade scale both for the antrum and the corpus [Five specimens from the antrum mucosa and five from the corpus mucosa were taken with medium-sized forceps. Two specimens were embedded in paraffin and the paraffin sections were stained using haematoxylin-eosin and Giemsa methods. The mucosal specimens were evaluated histologically according to the Sydney classification: presence of neutrophil infiltration, chronic lymphocytic inflammation, surface epithelial damage, atrophy, intestinal metaplasia, lymphoid follicles and e corpus -14.2 and diluted in the Brucella broth (Oxoid). H. pylori was isolated on the Columbia Agar Base supplemented with 7% horse blood and 1% Vitox (Oxoid) or Isovitalex (BBL). The plates were incubated for 3–7 days at 37°C under microaerobic conditions (CampyBak, BBL or CampyGen, Oxoid). H. pylori was identified by Gram staining and by oxidase, catalase and urease reactions [H. pylori strains to clarithromycin was estimated by E-test. The antibiotic cut-off points employed for the E-test were 1.0 mg/l (NCCLS, 2002).One specimen from the antrum and one from the corpus were placed in the Stuart Transport Medium (Oxoid) and taken to the laboratory within two hours for bacteriological examination. The biopsy samples were homogenised with sterile glass powder and under a stream of COeactions . The senH. pylori cells on the gastric mucosa and in the mucus layer.One specimen was used for imprinting the cytology slides from the antrum and corpus mucosa, fixed with 96% ethanol and stained by Acridine Orange (Difco, BBL) . The cytH. pylori from a frozen gastric biopsy specimen, a previously described procedure was used [glmM gene in each strain was established by PCR using primers, the reaction mixture, and thermal cycling [H. pylori NCTC 11637  and the DNA-free reaction mixture were assayed in separate tubes in each PCR and were run as the positive and negative controls of the reaction, respectively. The PCR products were identified by electrophoresis on 2% agarose gels.For DNA extraction of was used . The pre cycling ,20. DNA H. pylori was assessed positive if at least two tests were positive according to golden standard [standard .The data were analysed by Fisher's exact tests using the Jandel SigmaStat 2.0 program. Measurements from the GSRS were expressed as the mean values for dyspeptic syndrome.The study was carried out in accordance with the Helsinki Declaration and was approved by the Ethics Committee of the University of Tartu.Dyspeptic syndrome was found in all 34 cases. The mean GSRS score for the patients varied from 1.2 to 4.3.H. pylori infection in half of the investigated patients: positive 13C-UBT was found in 17 out of 34 cases. There was no difference between the mean GSRS score values for the H. pylori positive and negative cases (2.8 ± 1.8 vs. 2.9 ± 1.7, p > 0.05).The applied non-invasive test revealed H. pylori status and of the endoscopic finding are presented in Table On endoscopy, among the 34 patients, no ulcer or other mucosal defects were observed in 24 cases; erosions in the duodenal bulb were revealed in 6 cases and peptic ulcer was found in 4 cases . The data of H. pylori . The gastric and duodenal mucosa was visually normal in 11 H. pylori positive cases out of 17. On the contrary, only in 4 H. pylori positive cases did the endoscopic examination reveal the above mentioned peptic ulcers. Among the 6 cases with erosions of the duodenal mucosa, only two patients were H. pylori positive.A poor concordance was found between the visual examination of the gastric and duodenal mucosa on endoscopy and the applied non-invasive and invasive tests of H. pylori is shown in Table 13C-UBT and PCR were consistent with the data of histological examination both in 17H. pylori positive and 17 negative cases. On bacteriological examination, only one case, which was H. pylori positive both by PCR and the histological tests, was H. pylori negative. In contrast, cytological examination assessed typical H. pylori bacterial cells in only 4 of the 17H. pylori positive cases (24%), while all other cases (both positive and negative for H. pylori by the other methods) displayed abundant bacteria of different morphotypes.Comparison of the different diagnostic methods used for the detection of H. pylori was detected in 17 patients out of 34. Neutrophil infiltration, chronic inflammation, and surface epithelial damage both in the antrum and corpus mucosa were significantly expressed in the H. pylori positive cases (p < 0.001). Glandular atrophy and intestinal metaplasia were rarely observed both in the antrum and corpus mucosa of the H. pylori negative cases in comparison with the H. pylori positive cases, but the difference was not statistically significant (p > 0.05). Lymphoid follicles were more frequent in the antrum colonised with H. pylori (p < 0.05).The data of the histological findings are presented in Table H. pylori in 16 biopsy samples of the antral mucosa, while highly (> 256 mg/l) clarithromycin resistant H. pylori strains were found in 3 cases.Bacteriological investigation revealed H. pylori eradication therapy is of utmost importance for physicians as well for patients [H. pylori tests used before treatment, while post-treatment testing is not yet adequately studied. However, in the case of long-lasting recurrent dyspepsia after H. pylori eradication therapy, endoscopy has been strongly recommended [H. pylori status. Since persistent H. pylori positivity is always associated with possible peptic ulcer recurrence, the second line treatment according to bacterial susceptibility should be recommended. In the remaining cases where H. pylori is absent, the gastric mucosa is normal and no ulcer is detected, management of such patients should be aimed at establishment of other possible reasons for their complaints. Usually, a normal endoscopic finding reassures both the doctor and the patient [Proper diagnostic and therapeutic management of patients with dyspeptic syndrome after patients . Severalpatients ,23 have ommended . Our stu patient .H. pylori infection. On the contrary, our results demonstrate that although all patients with recurrent peptic ulcer were H. pylori positive, the minor visual findings in the other cases were not in concordance with H. pylori colonisation. Usually, the mucosa was visually normal even when H. pylori was found, and, on the contrary, most duodenal erosions occurred in H. pylori negative patients. The clinical data of our patients did not suggest earlier use of NSAID, which would have been one of the main reasons for H. pylori negative erosions. Therefore, after treatment, in presence of complaints, it is important to obtain samples for the investigation of gastric mucosa specimens to enhance the value of endoscopic examination. We completely agree with the opinion that the value of using mucosal specimens for histological evaluation of late post-treatment H. pylori eradication is sometimes underestimated [H. pylori test alone cannot solve the clinical problem of these patients. In our study, H. pylori negative patients had dyspeptic syndrome as well as gastric mucosal erosions, glandular atrophy and intestinal metaplasia. The last two lesions can presumably be associated with previous H. pylori infection and the follow-up of severe mucosal changes is recommended [H. pylori negative cases in accordance with endoscopic and histological findings.A recent study of Ohkusa et al.  showed tstimated . The nonommended . Hence iH. pylori infection even after treatment, and that the diagnostic value of a histology-based decision is high. Today, the value of mucosal specimens for the post-treatment histological diagnosis of H. pylori is considered low assuming that H. pylori colonisation may be patchy, or coccoid forms are difficult to detect [13C-UBT in parallel with histological investigation.Our study demonstrates that evaluation of the gastric mucosa with a focus on neutrophil and lymphocyte infiltration and epithelial damage is specific and sensitive for diagnosing o detect . We haveH. pylori completely correlates with the results of 13C-UBT and PCR both in H. pylori positive and negative cases. This confirms the validity of the histological evaluation of mucosal specimens in the case of recurrent peptic ulcer or erosions. Moreover, in countries with a high rate of H. pylori infection and gastric cancer, it is especially important to follow up patients for detecting dysplasia and malignancies [Next, for detecting the coccoid forms of the bacteria, we used additionally PCR method. Our results show that the histological finding of gnancies -29.H. pylori in cases where it was found by other methods. Cytology is highly evaluated for detection of H. pylori infection, as its agreement with histology is considered to be 100% [H. pylori detection, as different forms of the bacteria were found. The morphology of the helicobacters could have been modified for coccoid or otherwise non-typical forms. It is possible that some other bacteria might have colonised the mucosa due to reduced colonisation resistance after antibacterial treatment, failure of some intestinal functions or usage of medicines administered to relieve the feeling of discomfort [Surprisingly, brush cytology from the mucosa failed to detect  be 100% . Our resscomfort -33.H. pylori strains, which may result in the failure of repeat triple therapy. As the macrolide clarithromycin is chemically stable and well tolerated [Bacteriological investigation enabled to find a few clarithromycin resistant olerated , physiciH. pylori colonisation, PCR and bacteriology as they have shown good concordance with 13C-UBT. Moreover, endoscopy and histological investigation of a gastric biopsy have proved to be the methods with an additional diagnostic value, providing the physician with information about inflammatory, atrophic and metaplastic lesions of the stomach in dyspeptic H. pylori positive and negative patients. Bacteriological methods are suggested for detecting the putative antimicrobial resistance of H. pylori, aimed at successful eradication of infection in persistent peptic ulcer cases.In a late post-treatment setting, patients with dyspepsia should not be monitored only by non-invasive investigation methods; it is also justified to use the classical histological evaluation of The author(s) declare that they have no competing interests.13C-urea breath tests, and performed GSRS. KLõ carried out bacteriological examination. PH performed cytological examination. HA carried out molecular analysis and participated in the writing of the manuscript. HIM performed histological examination and statistical analysis, and participated in the design of the study and in the writing of the manuscript. MM coordinated the study and participated in the completion of the manuscript. All authors have read and approved the final version of the manuscript.HIM, IK and KLa carried out GSRS, endoscopy and gastrobiopsy. HK recruited patients, collected The pre-publication history for this paper can be accessed here:


However, when ThT assays were repeated using Aβ1-42, modest, but statistically significant, profibrillogenic activity was detected in both apolipoprotein E ε3- and apolipoprotein E ε4-containing media and was similar in magnitude for the two isoforms. These data demonstrate that native apolipoprotein E possesses "pathological chaperone"-type activity for Aβ: in other words, the data indicate that a chaperone-like misfolding reaction can occur between native apolipoprotein E and Aβ. However, the equipotent activities of the apolipoprotein E ε3 and ε4 isoforms suggests the possibility that either extended co-incubation of apolipoprotein E and Aβ, or, perhaps, the inclusion in the reaction of other fibrillogenesis-modulation co-factors  may be required for modeling in vitro the apolipoprotein E-isoform-specific-regulation of extracellular Aβ accumulation that occurs in vivo. Alternatively, other events, such as differential apolipoprotein E-isoform-mediated clearance of Aβ or of apolipoprotein E/Aβ complexes may underlie apolipoprotein E-isoform-dependent Aβ accumulation.The apolipoprotein E ε4 allele contributes to the genetic susceptibility underlying a large proportion (~40–60%) of typical, sporadic Alzheimer disease. Apolipoprotein E deficient mice made transgenic for human apolipoprotein E ε4 accumulate excess cerebral amyloid when compared to similarly prepared mice expressing human apolipoprotein E ε3. Therefore, it is important to search for relevant interactions(s) between apolipoprotein E ε4 and Aβ in order to clarify the biological role for apolipoprotein E ε4 in Alzheimer disease. Using a thioflavine T (ThT)-based assay, we have investigated the effects of native human apolipoprotein E isoforms on the kinetics of Aβ fibrillogenesis. No obvious profibrillogenic activity was detected in Aβ Using a thioflavine T (ThT)-based assay [Genetic-neuropathological correlation indicates that the apolipoprotein E type ε4 isoform specifies increased cerebral ,2 and ceed assay  we have 1-40 or Aβ1-42  was freshly prepared for each assay at a concentration of 16 mg/ml in distilled, deionized water and diluted with phosphate-buffered saline (PBS) to generate a 5 mg/ml working solution. The "aggregation step" consisted of a reaction mixture containing 8 μl Aβ peptide  and 32 μl of either apolipoprotein E ε3- or ε4-containing conditioned medium or control conditioned medium from SV40 empty vector-transfected cells.Synthetic AβFor the investigation of native apolipoprotein E preparations, apolipoprotein E isoforms were generated in the conditioned medium of stably-transfected SV40-apolipoprotein E ε3-, or SV40-apolipoprotein E ε4-, expressing Chinese hamster ovary (CHO) cells . All conditioned media were prepared using Dulbecco's minimal essential medium supplemented with 0.2% (wt/vol) bovine serum albumin only . Apolipoprotein E isoform levels were determined by quantitative immunoblotting of conditioned medium and apolipoprotein E-containing serum standards, the latter having been kindly provided by Dr. Petar Alaupovic of the Oklahoma Medical Research Foundation (Oklahoma City OK). Conditioned medium apolipoprotein E concentrations were then standardized using control medium conditioned by SV40 empty vector-transfected cells as diluent, yielding a final concentration of apolipoprotein E of 14 μg/ml, within the range of that reported in human cerebrospinal fluid. Since the final concentration of Aβ peptide was 1 mg/ml, the Aβ/apolipoprotein E stoichiometry (molar ratio) was ~500:1, suggesting models for the Aβ/apolipoprotein E interaction based either on a "catalytic" "pathological chaperoning" model of apolipoprotein E action on Aβ, or with a "seeding" model of Aβ folding. Detailed biochemical characterization of this native apolipoprotein E preparation has been reported .The "aggregation step" fibrillogenesis reaction  was incu1-40, either in physiological phosphate buffer alone or in the presence of metal ions , indicating that our Aβ preparations were capable of metal ion-induced fibrillogenesis. Metals were not present during assessment of profibrillogenic effects of apolipoprotein E isoforms.In order to standardize the ThT assay in our Laboratory, we performed studies of Aβ fibrillogenesis following 1–7 day incubations of Aβnal conc . ThT flunal conc   may be required for modeling in vitro the apolipoprotein E-isoform-specific-regulation of extracellular Aβ accumulation that occurs in vivo.These data demonstrate that native apolipoprotein E possesses "pathological chaperone"-type activity for Aβ: in other words, the data indicate that a chaperone-like misfolding reaction can occur between native apolipoprotein E and Aβ, at least at the concentrations and proportions evaluated herein. However, the equipotent activities of the apolipoprotein E ε3 and ε4 isoforms suggests the possibility that either extended co-incubation of apolipoprotein E and Aβ, or, perhaps, the inclusion in the reaction of other fibrillogenesis-modulation co-factors (such as metal ions, or inflammatory mediators such as reactive oxygen species, αin vivo.Alternatively, other events, such as differential apolipoprotein E-isoform-mediated clearance of Aβ or of apolipoprotein E/Aβ complexes ,9,14 mayThT, thioflavine T; Aβ, amyloid-β peptide; PBS, phosphate-buffered saline; CHO, Chinese hamster ovary cells.None declared.DS performed all assays, including the ThT assay, which was originated by HL. HL also oversaw the transfer of the assay from his lab to ours. RM prepared standard conditioned media from transfected cells provided by JDS. SG oversaw the project, supported the project as noted below, and wrote the manuscript.


The University of Edinburgh encourages its Ph.D. students to participate in a broad programme of science communication activities, designed to enhance public engagement in science The need for effective communication of research and the promotion of science is more important then ever. Public scepticism of research is high, and the number of students studying science continues to dwindle. In an attempt to combat this, the University of Edinburgh encourages its Ph.D. students to participate in a broad programme of science communication activities, designed to develop transferable skills they can use outside pure research and to enhance public engagement in science (PES).Included as part of the transferable skills programme, the University hosts a science communication module. The course provides an introduction to the different media, educational, ethical, and political issues surrounding the communication of science to a nonspecialist audience. Initially, the course addresses the presentation of science in written and oral forms. However, as the presentation of science to the public is not simply a practical skill, part of the course is dedicated to the tactical communication of science in society and the relationship between scientists and the media. Finally, students are introduced to ongoing PES opportunities. Any involvement requires the approval of supervisors, to ensure no adverse effects on academic performance. The activities available are diverse and flexible, and they enable students to undertake projects that reflect personal interest and availability.What opportunities are available? For enthusiastic students, membership of the Nikon–University of Edinburgh Post-Graduate Science Communication Team (PGSCT) is possible. The PGSCT requires a commitment of 15 days per academic year to paid science communication projects and events, including compulsory support of SCI-FUN, the University's established science and technology roadshow. The team is recruited annually from students of the Science and Engineering, Medicine, and Veterinary Medicine Colleges and takes a leading role in PES activities. Team members are supported by other graduate students who participate on a more casual basis.To ensure that a broad spectrum of activities is available to audiences, the PGSCT members are actively encouraged to design and develop their own ideas. An opportunity to do this is provided through the University's Science Zone, at the Royal Museum of Scotland, for the duration of the Edinburgh International Science Festival. From successful workshops, originally piloted at the science festival, several ex-PGSCT members have gone on to establish their own projects. One example of this is the Natural Environment Science Education scheme, which later received recognition through the Royal Society of Edinburgh/Scottish Executive ‘Science in the Community’ Award for 2003. A primary aim of this scheme was to deliver in isolated and remote communities innovative hands-on earth science- and natural science-based activities beyond city venues. For students, developing and presenting workshops are incredibly rewarding and allow them to experience the enthusiasm of participants and fellow presenters.Some activities are transferred from the Science Zone to the classroom, where students can communicate with children at all stages of their schooling. At primary school age (5–11 years) after-school science clubs, led by graduate students, provide an opportunity for presenters to share with pupils their enthusiasm for science. Alternative activities are directed at secondary school children (12–18 years), with a variety of workshops available across the different scientific disciplines. In particular, The Scottish Institute for Biotechnology Education (SIBE) has been set up within the University to facilitate PES activities such as the popular ‘Green Fingerprinting’ workshop where the principles of DNA fingerprinting are applied to an ecological scenario in the form of a practical activity . The majThe presentations may be in person or utilise new technologies such as video conferencing. A BBSRC Dialogue Award has recently been granted to SIBE to design, develop and deliver video-conferences addressing bio-ethical issues surrounding recent advances in biotechnology. Together, the two approaches of in-person and virtual presenting, enhance the schools' curriculum and facilitate dialogue between scientists and the public at a stage where promotion of science can influence the choice of further study and career options.The promotion of biotechnology within schools does not stop with the encouragement of school children's enthusiasm; SIBE also works closely with organisations such as ‘Science and Plants for Schools’ (SAPS) to assist with continuing professional development of biology teachers through the organisation of training days aimed at curriculum enhancement and the practical application of biotechnology in the classroom. Interested graduate students can co-present the workshops, though they are led by permanent staff. The students introduce their own research to highlight the applications of biotechnology and provide a useful technical resource.For students who prefer to communicate through the written word, though no guarantee of publication can be given, the University is in a position to highlight science-writing opportunities, usually as a contribution to publications specifically concerned with science communication or within the scientific press. At the Institute of Cell and Molecular Biology at the University a ‘press-gang’ meets on a monthly basis to discuss research carried out in the Institute and generate press releases for publication in the university press and national newspapers where appropriate.Complementing the University's efforts, many other organisations, such as UK Research Councils and the British Association for the Advancement of Science, endorse graduate students spending time on PES activities. Several schemes have been put in place to facilitate this; Researchers in Residence, the Science and Engineering Ambassadors Scheme and science communication courses. Hosted at the University's science campus—Kings Buildings—‘pgscicom’ provides up-to-date information on opportunities for PES at the University and beyond through regular email communications.As a PhD student and PGSCT member, I believe the approach of the University of Edinburgh to PES is of benefit to all involved. The combination of training and practical experience provides graduate students with new and valuable skills and opportunities to develop them further. The events and activities enthuse and engage audiences with the presentation of science in an informal but informative manner.


The GOLD.db (Genomics of Lipid-Associated Disorders Database) was developed to address the need for integrating disparate information on the function and properties of genes and their products that are particularly relevant to the biology, diagnosis management, treatment, and prevention of lipid-associated disorders. provides a reference for pathways and information about the relevant genes and proteins in an efficiently organized way. The main focus was to provide biological pathways with image maps and visual pathway information for lipid metabolism and obesity-related research. This database provides also the possibility to map gene expression data individually to each pathway. Gene expression at different experimental conditions can be viewed sequentially in context of the pathway. Related large scale gene expression data sets were provided and can be searched for specific genes to integrate information regarding their expression levels in different studies and conditions. Analytic and data mining tools, reagents, protocols, references, and links to relevant genomic resources were included in the database. Finally, the usability of the database was demonstrated using an example about the regulation of Pten mRNA during adipocyte differentiation in the context of relevant pathways.The GOLD.db The GOLD.db will be a valuable tool that allow researchers to efficiently analyze patterns of gene expression and to display them in a variety of useful and informative ways, allowing outside researchers to perform queries pertaining to gene expression results in the context of biological processes and pathways. The excessive consumption of high calorie, high fat diets and the adoption of a sedentary life style have made obesity and atherosclerosis major health problems in Western societies. In the USA, over 50% of the population are over-weight BMI > 25) and close to 25% are considered obese (BMI > 30) ,2. As a 5 and cloAt the root of obesity and atherosclerosis is an excessive deposition of neutral lipids. Adipose tissue accumulates predominantly triglycerides, whereas macrophages along the blood vessel wall mainly accumulate cholesterol and cholesteryl esters. Accordingly, a detailed understanding of the molecular mechanisms that govern the balance between lipid deposition and mobilization is fundamentally important for the prevention and improved treatment of disease. In addition to the apparent environmental components involved in the pathogenesis of disorders related to lipid and energy metabolism, a large number of studies have provided undisputed evidence that susceptibility genes contribute around 50% of the phenotype. These genes encode products involved in the cellular uptake, synthesis, deposition and/or mobilization of lipids. However, characterization of many if not most of these genes and their products remains rudimentary. Deficiencies in the current level of understanding extend to key enzymes such as important triglyceride hydrolases in adipose tissue  or choleWhile medical molecular biology traditionally associates single genes and gene products with diseases, a growing body of evidence suggests that several common disease phenotypes arise from the delicate interaction of many genes as well as gene-environment interactions. To elucidate the development of obesity and atherosclerosis, it will be necessary to analyze patterns of gene expression and relate them to various metabolic states. To discover novel genes, processes and pathways that regulate lipid deposition and mobilization, a departure from hypothesis-driven research and turn to a discovery-driven approach is necessary. The application of high-throughput technologies and genome-based analysis will provide the tools for the analysis of gene-gene and gene-environment interactions in a systematic and comprehensive manner.To facilitate genomic research we have initiated the development of a system for storing, integrating, and analyzing relevant data needed to decipher the molecular anatomy of lipid associated disorders. In order to provide a reference for pathways and information of the relevant genes and proteins in an efficiently organized way, we have created the Genomics Of Lipid-Associated Disorders database (GOLD.db). The GOLD.db integrates disparate information on the function and properties of genes and their protein products that are particularly relevant to the biology, diagnosis management, treatment, and prevention of lipid-associated disorders.The main goal of the GOLD.db was to provide biological pathways with image maps and visual pathway information. For each element in the pathway, specific information exists including structured information about a gene, protein, function, literature, and links. The GOLD.db provides also the possibility to map gene expression data individually to each pathway. Additionally, analytic and data mining tools, reagents, protocols, references, and links to relevant genomic resources were included in the database.The GOLD.db was implemented in Java technology . Hence, For tracking the repository of the reagents like clone resources and libraries which can be used for microarray studies, we have developed a relational database. Information about the vector, the sequence and length of the clone insert, primers for the PCR amplification, tissue, organism, accession number, library, container, storage information, date and person and access to other clone bases (e.g. IMAGE Consortium) can be stored. Users of the GOLD.db can list these clones and get all the information about each available clone. With restricted access, clone information or even clone lists can be uploaded and selection lists can be created and deleted. The input mask is designed in such way that the user can choose one of the elements of the created selection lists.In order to deal with the huge amount of data associated with large scale studies and to perform sequence based and microarray analysis, several bioinformatic tools were integrated or can be downloaded. Sequence similarity search against databases can be performed with BLAST  , FASTA  or HMM  to the corresponding elements of the available pathways similar to previous efforts . Either As key for the mapped relation the RefSeq number  is used. or ArrayExpress , it is still very difficult for researchers to obtain the original data. Web sites with Supplementary information are not maintained and/or not further developed. Hence, a database with a large collection of curated datasets will be enormously valuable for the community. Approaches to upload and retrieve gene expression data were pursued within the GOLD.db. Large scale gene expression data sets can be uploaded in form of tab delimited text files (Stanford file format) [Analysis of gene expression patterns in animal and cell models for lipid-associated disorders will help to understand the fundamental gene relations and regulatory mechanisms responsible for the development of obesity related diseases. The huge amount of data associated with the analysis of large scale gene expression analysis raises the demand of tools for storing, processing and retrieving complex information. Although a number of studies have been published and despite the requirements of some journals to deposit microarray data in public databases like GEO  format)  as used Recently, it was shown that insulin sensitivity, energy expenditure, and thermogenesis were enhanced in adipose-specific Pten-deficient (AdipoPten-KO) mice. Body and adipose tissues weight in these mice were significantly lower than those of control mice in spite of a larger food intake . We addrDuring adipocyte differentiation cyclin dependent kinase inhibitors, like p21 leads to a hypophosphorylation of the Retinoblastoma protein (Rb) which allows binding to the E2F transcription factor, causing cells to permanently exit the cell cycle – a required step in adipocyte differentiation called mitotic clonal expansion – before entering the terminal differentiation state. pRb interacts physically with adipogenic CCAAT/enhancer-binding proteins and positively regulates transactivation by C/EBPβ and therefore plays a pivotal role in adipocyte differentiation ,25. HencThus, using recently identified key player for food intake and weight control and using the GOLD.db, it is possible to address relevant questions and generate testable hypotheses on the molecular mechanisms of fat cell development.The vast quantity of gene expression data generated in genomic studies presents a number of challenges for their effective analysis and interpretation. In order to fully understand the changes in expression that will be observed, we must correlate these data with phenotype, genotype, metabolism and other information including the tissue distribution and time course expression data gleaned from previous studies. The goal of our work was the development of a specialized database and tools that allow researchers to efficiently analyze patterns of gene expression and to display them in a variety of useful and informative ways, allowing outside researchers to perform queries pertaining to gene expression results in the context of biological processes and pathways. The uniqueness of the GOLDdb database we have developed is threefold: 1) the inclusion of annotated pathways, 2) the availability of curated datasets and 3) the possibility to map experimental data on biological pathways. The upcoming challenges will be to include data from functional analysis and proteomics data, which will give us new opportunities in understanding mechanisms of different applications and lipid-associated disorders in particular.. The pathway editor and the clone tracker are available free of charge to academic, government, and other nonprofit institutions.The GOLD.db database should be cited with the present publication as a reference. Access to GOLD.db is possible through the world wide web at HH was responsible for the content, the annotation process, webdesign, and processing of data sets. MM was responsible for the implementation of the database and web application as well as the relational database for the clone tracker. BM and JH had implemented the mapping of expression data to pathways. GS is involved in providing of sequence analysis tools and server software. DMS has annotated the insulin signaling pathway. ZT was responsible for the design of the study and for overall project coordination.