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[Expectations of continuing medical education at first level of attention in Chiapas].

OBJECTIVE: To analyze the expectations of first level of attention physicians (FLAP) about continuing medical education. MATERIAL AND METHODS: An exploratory qualitative study was made from the cognitive view of motivation, in order to know the expectations of the FP at the Mexican Institute of Social Security (IMSS) in Chiapas. RESULTS: The FLAP show little interest in continuing medical education staff, because they find traditional professors and training courses off their work schedule, becoming difficult for them to attend. However, FLAP consider the library facilities and research activities as strategies to keep themselves updated, but they lack the required time to do so. CONCLUSIONS: The FLAP have low expectations caused by direct and indirect negative experiences at the work place as a result of ineffective communication among them and with the respective authorities of each unit. The FLAP show personal characteristics which limit their participation in the programs made by the continuing medical education staff, as well as their professional development.

Adult↗

Molecular subtyping of poultry-associated type A Clostridium perfringens isolates by repetitive-element PCR.

Clostridium perfringens strains (type A) isolated from an integrated poultry operation were subtyped using repetitive-element PCR with Dt primers. Isolates were obtained from fecal, egg shell, fluff, and carcass rinse samples as part of a previously reported temporally linked epidemiological survey. A total of 48 isolates of C. perfringens were obtained from different stages of the broiler chicken production chain from two separate breeder farms that supplied a single hatchery that in turn provided chicks to a single grow-out farm whose flocks were processed at a single plant. All 48 isolates were typeable (100% typeability) by repetitive-element PCR with Dt primers. This subtyping method was highly reproducible and discriminatory. By repetitive-element PCR with Dt primers, isolates were classified into four major branches with 12 subgroups or clades. The Simpson's index of discrimination was calculated to be 0.96 for groupings of >95% correlation. Toxin gene profiles of the isolates indicated that all of the isolates were C. perfringens alpha-toxin gene positive and 46 of 48 isolates were beta2-toxin gene positive. All strains were negative for beta- and epsilon-toxin genes. Repetitive sequence-based PCR was found to be a technically practical and reproducible means of subtyping C. perfringens libraries from specific epidemiological or production environment settings.

Animals↗

Quantitative analysis of a RNA-cleaving DNA catalyst obtained via in vitro selection.

In vitro selections performed in the presence of Mg(2+) generated DNA sequences capable of cleaving an internal ribonucleoside linkage. Several of these, surprisingly, displayed intermolecular catalysis and catalysis independent of Mg(2+), features that the selection protocol was not explicitly designed to select. A detailed physical organic analysis was applied to one of these DNAzymes, termed 614. First, the progress curve for the reaction was dissected to identify factors that prevented the molecule from displaying clean first-order transformation kinetics and 100% conversion. Several factors were identified and quantitated, including (a) competitive intra- and intermolecular rate processes, (b) alternative reactive and unreactive conformations, and (c) mutations within the catalyst. Other factors were excluded, including "approach to equilibrium" kinetics and product inhibition. The possibility of complementary strand inhibition was demonstrated but was shown to not be a factor under the conditions of these experiments. The rates of the intra- and intermolecular processes were compared, and saturation models for the intermolecular process were built. The rate-limiting step for the intermolecular reaction was found to be the association/folding of the enzyme with the substrate and not the cleavage step. The DNAzyme 614 is more active in trans than in cis and more active at temperatures below the selection temperature than at the selection temperature. Many of these properties have not been reported in similar systems; these results therefore expand the phenomenology known for this class of DNA-based catalysts. A brief survey of other catalysts arising from this selection found other Mg(2+)-independent DNAzymes and provided a preliminary view of the ruggedness of the landscape, relating function to structure in sequence space. Hypotheses are suggested to account for the fact that a selection in the presence of Mg(2+) did not exploit this Mg(2+). This study of a specific catalytically active DNAzyme is an example of studies that will be necessary generally to permit in vitro selection to help us understand the distribution of function in sequence space.

Bacteriophage lambda↗

"Mini-Medical School for Librarians": from needs assessment to educational outcomes.

PURPOSE: This study evaluates the outcomes of the "Mini-Medical School for Librarians" or "Medical School Experience," a continuing education symposium designed to improve librarians' understanding of medicine and medical education. SUBJECTS: The subjects are the symposium participants, a group that consisted of fifty-eight medical librarians and other information professionals. METHODOLOGY: Pre- and post-symposium self-evaluation surveys gauged participants' self-assessed confidence with the course content. A follow-up survey was administered six months after the symposium. A learning action plan recorded both the intended and actual applications of course content to professional settings. RESULTS: T-test analysis of paired pre- and post-symposium responses reveal a significant positive change in the mean self-assessed confidence with course content immediately following the symposium. Pairings of post-symposium and follow-up survey responses indicate a slight reversal in attendees' confidence in the months following the symposium, but pairings of pre-symposium and follow-up survey results demonstrate that the longitudinal impact of the program on self-assessed confidence with course content was positive and significant. Analysis of the learning action plan revealed a disparity in how participants planned to use the information they learned in the course and how they actually used it. CONCLUSIONS: Continuing education programs that address the content and structure of medicine can be an effective means by which to inform both the novice's and mid-career medical librarian's understanding of medicine and medical education.

Education, Continuing↗

Genetics of palate development.

We are beginning to see a tantalizing picture by looking into the "genetic window" on the teratogenesis of cortisone-induced cleft palate and on normal palate development. We must continue to open this window. So far, all strains of mice that have been treated with cortisone do react with induced cleft palate and, from their dose-response behaviors, they appear to react in the same way. It is tempting to conclude that dosage tolerance is the only genetic variation in the reaction. We do not attach any significance to this because we must emphasize the fact that these different strains represent an insignificant sample of the hundreds of genetically different, recognized strains of the mouse. We have emphasized previously that systematic strain surveys must become an active part of teratological genetics in order to obtain an approximation of how much genetic variation exists in a trait in the species (e.g., Biddle, 1981). Now, we need to go beyond strain surveys. Crosses between strains must become part of this activity. Dominance appears to be a property of the homeostatic model of palate development as far as the cortisone probe is concerned, but it may not be a universal property. The significance of dominance and the architecture of gene action can only be interpreted by exploring more strain pairs in the mouse. A study of two or three or four strains is simply inadequate. From the crosses between a limited number of strains, major genetic factors in cortisone reactivity have been identified. These major genetic factors may regulate the homeostatic model of palate development, but we do not know this for a fact. The identification of major genetic factors has moved us to a new horizon; we can move now from a purely descriptive biology to a study of the functional biology of the process of palate development. This move will require not only the molecular probes with which to walk along the specific chromosomes to the major genetic factors but also a library of fully characterized strains of the mouse in which to use these probes. The library of strains that have been genetically characterized for the cleft palate reaction is missing.

Animals↗

Isolation and tissue profiles of a large panel of phage antibodies binding to the human adipocyte cell surface.

Phage display is a powerful technique for the rapid selection and isolation of antibodies to any given target antigen. We have applied this technology to isolate over 100 different human antibodies that bind to antigens expressed in situ on the human adipocyte cell surface. This is a diverse panel of antibodies, as indicated by the V-region sequences. The binding profile of each anti-adipocyte antibody has been characterised using phage antibody immunocytochemistry against a panel of normal human tissues. Although there was some variation in the intensity of the adipocyte staining, each antibody consistently recognised adipocytes, where present, irrespective of the tissue source. In addition, all of the antibodies recognised at least one other cell type other than the adipocyte cell surface. In total, over 50 different tissue-binding profiles were recorded, with the most frequently recognised tissues identified as capillaries or smooth muscle. Extensive tissue binding profiles were generated for some antibodies using a panel of 37 different human tissues. This identified anti-adipocyte antibodies with unexpected profiles, such as FAT.13, which binds only to adipocytes and capillaries in the entire tissue panel. We believe this is the most extensive survey ever undertaken of the human adipocyte cell surface. Moreover, similar methodology could be used to derive complete tissue-binding profiles of antibodies against cell-surface antigens of any cell type. Indeed, by screening antibodies on both normal and diseased tissues, it may be possible to identify antigenic associations between different cell types and the pathologies of many diseases.

Adipocytes↗

Hepatitis B vaccine boosters: is there a clinical need in high endemicity populations?

The Steering Committee for the Prevention and Control of Infectious Diseases in Asia recently conducted a survey of primary-care physicians in Asia, which revealed that many physicians administer boosters in their clinical practice and that there is considerable variation and uncertainty among physicians regarding this practice. This paper serves as a response to physicians' uncertainties by reviewing the literature regarding the administration of hepatitis B vaccine boosters in high endemicity areas and presenting the Steering Committee's guidelines for booster administration. While there are few data to support a need for routine hepatitis B vaccine boosters as a public health measure, they help to provide reassurance of immunity against breakthrough infection in certain risk groups. In clinical practice, primary-care physicians must exercise their judgment regarding the need for booster vaccination on an individual basis. This paper examines the available literature on the administration and value of hepatitis B vaccine boosters, explores the differences between the public health approach and clinical practice, and provides guidelines for those who use boosters in high endemicity Asian populations. Relevant articles were identified through searches of MEDLINE (1975-2003) and the Cochrane Library, using 'hepatitis B' and 'booster' as primary search terms. Guidelines for those who decide to administer hepatitis B vaccine boosters include: boosting approximately 10-15 years after primary vaccination; boosting rather than not when monitoring of antibody levels is not feasible; boosting immunocompromised patients when the antibody to hepatitis B surface antigen titer falls below 10 mIU/mL; and boosting healthcare workers based on the endemicity of the particular country.

Endemic Diseases↗

Frequencies and sequence characteristics of di-, tri-, and tetra-nucleotide microsatellites in wheat.

Microsatellites have emerged as an important source of genetic markers for eukaryotic genomes. In this report, two wheat (Triticum aestivum L.) genomic libraries were screened for several di-, tri-, and tetranucleotide tandem repeats. Clones containing (AC)n, (AG)n, (TCT)n, and (TTG)n repeats were isolated and sequenced. On average, there was one (AC)n microsatellite every 292 kbp and one (AG)n microsatellite every 212 kbp. The trinucleotide tandem repeats (TCT)n and (TTG)n were about 10 times less common than the two dinucleotide tandem repeats tested and tetranucleotide tandem repeats were rare. Many of the microsatellites had more than 10 repeats. The maximum repeat number found for (AC)n was 36 and for (TCT)n was more than 50. The prevailing category of (AG)n microsatellites from (AG)n isolates was perfect repeats. About half of the (AC)n microsatellites were compound repeats, while most of the (TCT)n microsatellites were imperfect repeats. In a small sample, (TTG)n microsatellites consisted mainly of compound repeats. The most frequently associated repeats were (AC)n with (AG)n, (TCT)n with (TCC)n, and (TTG)n with (TGG)n. Among 32 pairs of microsatellite primers surveyed, seven produced polymorphic products in the expected size range and these loci were mapped using a hexaploid wheat mapping population or aneuploid stocks.

Base Sequence↗

Expression of cone transducin, Gz alpha, and other G-protein alpha-subunit messenger ribonucleic acids in pancreatic islets.

The G-proteins are a family of heterotrimeric guanine nucleotide-binding proteins that play important roles in signal transduction and whose expression is regulated in a tissue-specific manner. Here we have surveyed the expression of G-protein alpha-subunits in mouse pancreatic islets. Degenerate oligonucleotide primers corresponding to conserved primary sequences in known G alpha-subunits were used in a reverse transcriptase-polymerase chain reaction, and the amplified complementary DNA (cDNA) fragments were subcloned and sequenced. Over 100 clones were analyzed, from which we determined that islet cells express at least seven G alpha-subunits: G8 alpha, Gi1 alpha or Gi3 alpha, Gi2 alpha, G11 alpha, G14 alpha, Gz alpha, and Gt2 alpha (cone transducin). In particular, the identification of Gz alpha and Gt2 alpha was of interest in that previous studies had indicated that the expression of Gz alpha was restricted mainly to the brain, retina, and adrenal gland, whereas Gt2 alpha was expressed predominantly in retinal cone photoreceptors. By Western blot analysis, we estimated that the amount of Gz alpha protein present in mouse islets was about 40% of that in retina. To further investigate the expression of Gt2 alpha, mouse Gt2 alpha cDNA was cloned from a retinal library and sequenced. The cDNA was used as a probe for Northern blot analysis, and the results confirmed that mouse islets contained a substantial level of Gt2 alpha messenger RNA (mRNA), albeit less than that found in retina (approximately 5-fold lower). Gt2 alpha mRNA was also shown to be present in a clonal mouse pancreatic alpha-cell line (alpha TC1-6) as well as in adrenal gland, pituitary, and a clonal mouse anterior pituitary cell line (AtT20). In situ hybridization revealed that Gt2 alpha mRNA was expressed essentially throughout the islet, suggesting that it is normally expressed in the abundant islet beta-cells and possibly others. In situ analysis also showed that Gt2 alpha mRNA expressed in the pituitary was limited to the intermediate and anterior lobes. We conclude that islet cells express multiple G-proteins, including several that are normally expressed at high levels in certain neuronal cells.

Amino Acid Sequence↗

Health information on the internet: patient empowerment or patient deceit?

Internet is the biggest medical library in the world. It has transformed the way many health seekers find health information. Seekers on net have exponentially increased from 54 million in 1998 to 110 million in 2002 (U.S. figures) and are ever increasing. Act of looking for health or medical information is the third most popular activities online. Search engines are used by almost 81% of the e-patients to look for the information they want. Internet is fast becoming an influential force as more than 70% consumers say that the information on the net has influenced their treatment decisions. However, the reliability of all the health information available on the Internet is questionable. Numerous studies have indicated deficiencies in the quality of information on the Internet. Due to technical and commercial reasons, the results provided by various search engines can be potentially biased. Only about a quarter of health seekers thoroughly check the source, timeliness of information every time they search for health information. In fact, most adults from USA, Japan, France and Germany who participated in a recent survey thought online health care information to be trustworthy, of good quality, easy to understand and easy to find. Efforts are now being taken to ensure the quality of health information on the Internet. Patients need to be educated about the worthiness of a site and also be prescribed the right sites to be consulted for information.

Health Education↗

Dentocase - open-source education management system in dentistry.

Since 2001, an interdisciplinary project on multimedia education in medicine has been sponsored by the Federal Ministry of Education and Research at the Charité. One part of the project is on dentistry. In the light of the results of a survey of dental students, an Internet-based education management system was created using open-source back-end systems. It supports four didactic levels for editing documentation of patient treatments. Each level corresponds to the learning abilities of the students. The patient documentation is organized to simulate the working methods of a physician or dentist. The system was tested for the first time by students in the summer semester of 2003 and has been used since the winter semester of 2003 as part of the curriculum.

Computer-Assisted Instruction↗

Novel gene expression profiles define the metabolic and physiological processes characteristic of wood and its extractive formation in a hardwood tree species, Robinia pseudoacacia.

Wood is of critical importance to humans as a primary feedstock for biofuel, fiber, solid wood products, and various natural compounds including pharmaceuticals. The trunk wood of most tree species has two distinctly different regions: sapwood and heartwood. In addition to the major constituents, wood contains extraneous chemicals that can be removed by extraction with various solvents. The composition and the content of the extractives vary depending on such factors as, species, growth conditions, and time of year when the tree is cut. Despite the great commercial and keen scientific interest, little is known about the tree-specific biology of the formation of heartwood and its extractives. In order to gain insight on the molecular regulations of heartwood and its extractive formation, we carried out global examination of gene expression profiles across the trunk wood of black locust (Robinia pseudoacacia L.) trees. Of the 2,915 expressed sequenced tags (ESTs) that were generated and analyzed in the current study, 55.3% showed no match to known sequences. Cluster analysis of the ESTs identified a total of 2278 unigene sets, which were used to construct cDNA microarrays. Microarray hybridization analyses were then performed to survey the changes in gene expression profiles of trunk wood. The gene expression profiles of wood formation differ according to the region of trunk wood sampled, with highly expressed genes defining the metabolic and physiological processes characteristic of each region. For example, the gene encoding sugar transport had the highest expression in the sapwood, while the structural genes for flavonoid biosynthesis were up-regulated in the sapwood-heartwood transition zone. This analysis also established the expression patterns of 341 previously unknown genes.

DNA, Complementary↗

The role of expert searching in the Family Physicians' Inquiries Network (FPIN).

OBJECTIVE: This article describes the contributions of medical librarians, as members of the Family Physicians' Inquiries Network (FPIN), to the creation of a database of clinical questions and answers that allows family physicians to practice evidence-based medicine using high-quality information at the point of care. The medical librarians have contributed their evidence-based search expertise and knowledge of information systems that support the processes and output of the consortium. METHODS: Since its inception, librarians have been included as valued members of the FPIN community. FPIN recognizes the search expertise of librarians, and each FPIN librarian must meet qualifications demonstrating appropriate experience and training in evidence-based medicine. The consortium works collaboratively to produce the Clinical Inquiries series published in family medicine publications. RESULTS: Over 170 Clinical Inquiries have appeared in Journal of Family Practice (JFP) and American Family Physician (AFP). Surveys have shown that this series has become the most widely read part of the JFP Website. As a result, FPIN has formalized specific librarian roles that have helped build the organizational infrastructure. CONCLUSIONS: All of the activities of the consortium are highly collaborative, and the librarian community reflects that. The FPIN librarians are valuable and equal contributors to the process of creating, updating, and maintaining high-quality clinical information for practicing primary care physicians. Of particular value is the skill of expert searching that the librarians bring to FPIN's products.

Databases as Topic↗

Health information in Italian public health websites: moving from inaccessibility to accessibility.

AIMS: In January 2004, the Italian government passed new legislation designed to give people with disabilities access to online services. Starting from the new legislation requirements, the purpose of this study is to investigate whether Italian Local Health Authorities websites are accessible to users with different abilities. METHODS: One hundred and seventy websites were analysed evaluating their compliance to the World Wide Web Consortium (W3C) Priority 1 guidelines via a combination of appropriate accessibility testing methods. This review was carried out twice: the first time it took place between the end of 2003 and May-June 2004. The same survey was repeated again in December 2004, after the issuing of the draft of the Implementation Regulations for The Stanca Law in July 2004. In this second survey, all the websites analysed before were checked again to establish if some measures to make those sites accessible were taken. RESULTS: Seventy-six per cent of the analysed websites fail to satisfy the most basic W3C requirements, but Italian Local Health Authorities websites are likely to present significant improvements to access for some disabled user groups: the number of accessible websites increased consistently between the first and the second review. CONCLUSIONS: It seems that awareness of web accessibility issues is increasing among developers of health information websites because of law enforcement. Recommendations for future research and development are offered.

Consumer Behavior↗

A multiple PCR-primer approach to access the microeukaryotic diversity in environmental samples.

The Cariaco Basin off the Venezuelan coast in the Caribbean Sea is the world's largest truly marine body of anoxic water. The first rRNA survey of microbial eukaryotes in this environment revealed a number of novel lineages, but sampled only a fraction of the entire diversity. The goal of this study was to significantly improve recovery of protistan rRNA from the Basin. This was achieved by a systematic application of multiple PCR primer sets and substantially larger sequencing efforts. We focused on the most diverse habitat in the basin, anoxic waters approximately 100m below the oxic-anoxic interface, and detected novel lineages that escaped the single PCR primer approach. All clones obtained proved unique. A 99% sequence similarity cut-off value combined these clones into operational taxonomic units (OTUs), over 75% of which proved novel. Some of these OTUs form deep branches within established protistan groups. Others signify discovery of novel protistan lineages that appear unrelated to any known microeukaryote. Surprisingly, even this large-scale multi-primer rRNA approach still missed a substantial part of the samples' rRNA diversity. The overlap between the species lists obtained with different primers is low, with only 4% of OTUs shared by all three libraries, and the number of species detected only once is large (55%). This strongly indicates that, at least in anoxic environments, protistan diversity may be much larger than is commonly thought. A single sample appears to contain thousands of largely novel protistan species. Multiple PCR primer combinations may be needed to capture these species.

Animals↗

Aeromonas salmonicida possesses two genes encoding homologs of the major outer membrane protein, OmpA.

Two homologs of the outer membrane protein OmpA were identified in Aeromonas salmonicida by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, and amino-terminal sequence analyses. An A. salmonicida genomic DNA library was constructed by using lambda GEM-11 and recombinant phage carrying both genes ompAI and ompAII) selected by immunoscreening. A 5.0-kb BamHI fragment containing the two genes in tandem was subcloned in pBluescript and used for further subcloning and sequencing of the genes. The encoded proteins (Mr = 33,564 and 32,536 for mature OmpAI and OmpAII, respectively) had only 64% identity with each other and otherwise had the highest level of homology to OmpA proteins from the members of the family Enterobacteriaceae. Based on the Escherichia coli OmpA model, an eight-stranded amphipathic beta-barrel model for the membrane assembly of the N-terminal half of OmpAI and OmpAII was predicted. Most variation between the two proteins was localized to the predicted surface loops and periplasmic turns, while the transmembrane strands and C-terminals domains were highly conserved. Expression of ompAI and ompAII separately in E. coli indicated that both genes could be independently transcribed from their own promoters and that both gene products were assembled into the E. coli outer membrane. A survey of different Aeromonas spp. by PCR revealed that possession of two tandem ompA genes was widespread among this genus. This is the first report of any bacterial species possessing two genes for homologs of this major outer membrane protein.

Aeromonas↗

Nucleotide variation at the Gpdh locus in the genus Drosophila.

The Gpdh locus was sequenced in a broad range of Drosophila species. In contrast to the extreme evolutionary constraint seen at the amino acid level, the synonymous sites evolve at rates comparable to those of other genes. Gpdh nucleotide sequences were used to infer a phylogenetic tree, and the relationships among the species of the obscura group were examined in detail. A survey of nucleotide polymorphism within D. pseudoobscura revealed no amino acid variation in this species. Applying a modified McDonald-Kreitman test, the amino acid divergence between species in the obscura group does not appear to be excessive, implying that drift is adequate to explain the patterns of amino acid change at this locus. In addition, the level of polymorphism at the Gpdh locus in D. pseudoobscura is comparable to that found at other loci, as determined by a Hudson-Kreitman-Aguadé test. Thus, the pattern of nucleotide variation within and between species at the Gpdh locus is consistent with a neutral model.

Algorithms↗

A novel complementary deoxyribonucleic acid is abundantly and specifically expressed in the uterus during pregnancy.

OBJECTIVE: Our purpose was to identify novel genes expressed by the uterus during late pregnancy. STUDY DESIGN: A complementary deoxyribonucleic acid library constructed from late pregnancy mouse uterus was screened by differential hybridization with complementary deoxyribonucleic acid probes constructed from late pregnancy mouse uterus and nonpregnant mouse uterus. Radiolabeled complementary deoxyribonucleic acid probes derived from one of the complementary deoxyribonucleic acids isolated were used in northern hybridizations against ribonucleic acid collected from pregnant and nonpregnant uterus and a variety of other mouse tissues. RESULTS: A total of 40 positive clones were isolated; half were identified as cytotoxic T-lymphocyte antigen-2 alpha (a putative inhibitor of the protease cathepsin L) and the other half represented a novel complementary deoxyribonucleic acid. Conceptual translation of the complementary deoxyribonucleic acid predicted a novel protein of 154 amino acids that is proline rich and acidic (pregnancy-specific uterine protein). Northern hybridizations demonstrated that message is abundant in the uterus during late pregnancy. After birth expression rapidly decreased and message was no longer found in the uterus by the third day. A minimal amount of message is present in placental ribonucleic acid, but expression is otherwise not detected in a variety of adult and fetal tissues surveyed, suggesting that expression of this gene is limited to the pregnant uterus. CONCLUSIONS: The abundance of message and expression apparently limited to the pregnant uterus suggests that the protein represented by this complementary deoxyribonucleic acid may play an important role in pregnancy.

Animals↗