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PubMed2000

Feverfew for preventing migraine.

BACKGROUND: Feverfew (Tanacetum parthenium L.) is a popular herbal remedy for migraine. OBJECTIVES: To systematically review the evidence for or against the efficacy of feverfew versus placebo for the prevention of migraine. SEARCH STRATEGY: Electronic literature searches were performed using the databases CISCOM (Research Council for Complementary Medicine, London, UK), MEDLINE, EMBASE, Biosis and the Cochrane Library (each from its inception to April 1998). Manufacturers were contacted and the bibliographies of identified articles checked for further trials. SELECTION CRITERIA: Randomised, placebo-controlled, double-blind trials assessing the efficacy of feverfew for preventing migraine were included. No restrictions regarding the language of publication were imposed. DATA COLLECTION AND ANALYSIS: Data on patients, interventions, methods, outcomes and results were extracted in a pre-defined, standardised manner. Methodological quality was evaluated using the scoring system developed by Jadad and colleagues. Both data extraction and the assessment of methodological quality were performed independently by two reviewers. MAIN RESULTS: Four trials met the inclusion criteria. The majority of these trials suggested beneficial effects of feverfew compared with placebo. However, the trial with the highest methodological quality, which was also among the largest, found no significant difference between feverfew and placebo. REVIEWER'S CONCLUSIONS: The efficacy of feverfew for the prevention of migraine has not been established beyond reasonable doubt.

Complementary Therapies↗
PubMed2005

School-based prevention for illicit drugs' use.

BACKGROUND: Drug addiction is a chronic, relapsing disease. Primary interventions should be aimed to reduce first use, or prevent the transition from experimental use to addiction. School is the appropriate setting for preventive interventions. OBJECTIVES: To evaluate the effectiveness of school-based interventions in improving knowledge, developing skills, promoting change, and preventing or reducing drug use versus usual curricular activities or a different school-based intervention . SEARCH STRATEGY: MEDLINE , EMBASE, ERIC, PSYCHINFO, Cochrane Library, ACP Journal Club, Cochrane Drug and Alcohol Group Register, updated to February 2004, were searched. Bibliography of papers was checked and personal contacts were made to identify other relevant studies. SELECTION CRITERIA: RCTs, CCTs or Controlled Prospective Studies (CPS) evaluating school-based interventions designed to prevent substance use. DATA COLLECTION AND ANALYSIS: Data were selected and extracted independently by two reviewers. Quality was assessed with the CDAG checklist. Interventions were classified as skills, affective, knowledge-focused and other characteristics were also studied (teaching, follow-up implementation, context activation). MAIN RESULTS: 32 studies (29 RCTs and 3 CPSs) were included. 28 were conducted in the USA; most were focused on 6th-7th grade students, and based on post-test assessment. RCTs: (1) Knowledge vs usual curricula: Knowledge focused programs improve drug knowledge (SMD=0.91; 95% CI: 0.42, 1.39).(2) Skills vs usual curricula: Skills based interventions increase drug knowledge (WMD=2.60; 95% CI: 1.17-4.03), decision making skills (SMD=0.78; CI95%: 0.46-1.09), self-esteem (SMD=0.22; CI95%: 0.03-0.40), peer pressure resistance (RR=2.05; CI95%: 1.24-3.42), drug use (RR=0.81; CI95%: 0.64, 1.02), marijuana use (RR=0.82; CI95%: 0.73, 0.92) and hard drug use (RR=0.45; CI95%: 0.24-0.85). (3) Skills vs knowledge: No differences are evident.(4) Skills vs affective: Skills-based interventions are only better than affective ones in self-efficacy (WMD=1.90; CI95%: 0.25, 3.55). (5) Affective vs usual curricula: Affective interventions improve drug knowledge (SMD=1.88; CI95%: 1.27, 2.50) and decision making skills (SMD=1.35; CI95%: 0.79, 1.9). (6) Affective vs knowledge: Affective interventions improve drug knowledge (SMD=0.60; CI95%: 0.18,1.03), and decision making skills (SMD=1.22; CI95%: 0.33, 2.12). Results from CPSs: No statistically significant results emerge from CPSs. AUTHORS' CONCLUSIONS: Skills based programs appear to be effective in deterring early-stage drug use. The replication of results with well designed, long term randomised trials, and the evaluation of single components of intervention (peer, parents, booster sessions) are the priorities for research. All new studies should control for cluster effect.

Adolescent↗
PubMed2005

Antioxidants for preventing pre-eclampsia.

BACKGROUND: Oxidative stress has been proposed as a key factor involved in the development of pre-eclampsia. Supplementing women with antioxidants during pregnancy may help to counteract oxidative stress and thereby prevent or delay the onset of pre-eclampsia. OBJECTIVES: To determine the effectiveness and safety of any antioxidant supplementation during pregnancy and the risk of developing pre-eclampsia and its related complications. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group Trials Register (June 2004) and the Cochrane Central Register of Controlled Trials (The Cochrane Library, Issue 3, 2004). SELECTION CRITERIA: All randomised and quasi-randomised trials comparing one or more antioxidants with either placebo or no antioxidants during pregnancy for the prevention of pre-eclampsia, and trials comparing one or more antioxidants with another, or with other interventions. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trials for inclusion, data extraction and trial quality. Data were double-entered into the Review Manager software. MAIN RESULTS: Seven trials involving 6082 women are included in this review. The largest trial (5021 women) was quasi-random and only three of the seven included trials were rated high quality. Supplementing women with any antioxidants during pregnancy compared with control or placebo was associated with a 39% reduction in the risk of pre-eclampsia (relative risk (RR) 0.61, 95% confidence intervals (CI) 0.50 to 0.75, seven trials, 6082 women). Women receiving antioxidants compared with control or placebo also had a reduced risk of having a small-for-gestational-age infant (RR 0.64, 95% CI 0.47 to 0.87, three trials, 634 women), their infants had a greater mean birthweight (weighted mean difference 91.83 g, 95% CI 11.55 to 172.11, three trials, 451 women), but they were more likely to give birth preterm (RR 1.38, 95% CI 1.04 to 1.82, three trials, 583 women). There were insufficient data for reliable conclusions about possible effects on any other outcomes. AUTHORS' CONCLUSIONS: These results should be interpreted with caution, as most of the data come from poor quality studies. Nevertheless, antioxidant supplementation seems to reduce the risk of pre-eclampsia. There also appears to be a reduction in the risk of having a small-for-gestational-age baby associated with antioxidants, although there is an increase in the risk of preterm birth. Several large trials are ongoing, and the results of these are needed before antioxidants can be recommended for clinical practice.

Antioxidants↗
PubMed2005

Direct amplification of rhizobial nodC sequences from soil total DNA and comparison to nodC diversity of root nodule isolates.

A group-specific primer set was developed using nodC as a target gene for the amplification of rhizobial sequence diversity from nodule isolates and total soil DNA preparations. The primer set was tested on 209 nodule isolates, recovered from six different trap plant species which were grown in two soil samples collected from a chickpea and a wheat field site in India. We also amplified and cloned PCR products from total DNA isolated from the same soil samples. The total diversity within the resulting clone libraries (Sigma 218 clones) was higher than that recovered from trap plants, but differed depending on the PCR protocols and primers used. However, some plant-selected genotypes could not be obtained using the community approach, probably due to variable detection limits and limited clone library sizes.

Bacterial Proteins↗
PubMed2002

Prediction of drug solubility from structure.

The aqueous solubility of a drug is an important factor affecting its bioavailability. Numerous computational methods have been developed for the prediction of aqueous solubility from a compound's structure. A review is provided of the methodology and quality of results for the most useful procedures including the model implemented in the QikProp program. Viable methods now exist for predictions with less than 1 log unit uncertainty, which is adequate for prescreening synthetic candidates or design of combinatorial libraries. Further progress with predictive methods would require an experimental database of highly accurate solubilities for a large, diverse collection of drug-like molecules.

Chemical Phenomena↗
PubMed2003

Combined electrospray ionization-atmospheric pressure chemical ionization source for use in high-throughput LC-MS applications.

Fast and accurate analytical methods are essential to keep pace with sample libraries produced from combinational chemistry and high-throughput biological screening. Many laboratories now use a combination of ionization techniques for the characterization of these samples, including atmospheric pressure chemical ionization (APCI), electrospray ionization (ESI), and photoionization (PI). Data are shown here from the analysis of a compound collection plate containing a variety of sample structures. ESI will normally analyze around 80% of these samples, necessitating a source change to analyze a further 10%. In this work, we have developed a new combined ESI-APCI source (ESCi) for use in on-line HPLC applications. The combined source allows alternate on-line ESI and APCI scans with polarity switching within a single analysis. The ESCi source has been designed to be a simple replacement for the existing mass spectrometer interfaces. Each ionization method is optimized independently using separate tuning parameters. Instrument electronics can readily switch between the two ionization methods and polarities within normal interscan time periods. The new source has reduced the analysis time of sample plates by eliminating the need for a source hardware change, source optimization, and repeat analyses.

Journal Article↗
PubMed2006

Guidelines for the management of pelvic pain associated with endometriosis: a systematic appraisal of their quality.

BACKGROUND: Guidelines exist for the management of endometriosis. Validated and reliable appraisal tools exist to assess the quality of guidelines. OBJECTIVES: To systematically appraise the quality of guidelines for the management of pelvic pain associated with endometriosis. SEARCH STRATEGY: Guidelines were identified using a prospective protocol through a systematic search of MEDLINE (1951-2005), EMBASE (1974-2005), the Cochrane Library (2005, issue 2), known guideline websites and the World Wide Web. SELECTION CRITERIA: Type of document: guideline, consensus statement, care protocol or healthcare technology assessment produced by national or international professional organisations and societies or governmental agencies; subject: management of pelvic pain associated with endometriosis. DATA COLLECTION AND ANALYSIS: Two validated appraisal tools, Cluzeau and The Appraisal of Guidelines and Research and Evaluation (AGREE), were used to quantitatively assess the quality of guidelines. Areas evaluated included 'rigour of development', 'context and content' and 'application.' MAIN RESULTS: Eight of 596 potentially relevant citations identified met our inclusion criteria. The Cluzeau instrument quality score were the following: rigour of development, 53% (range 5-65%); context and content, 69% (range 29-79%) and application 20% (range 0-20%). The application dimension achieved significantly lower quality scores (P = 0.026 versus rigour of development and P = 0.017 versus context and content). The AGREE instrument quality scores were the following: scope and purpose, 68% (range 17-89%); stakeholder involvement, 33% (range 13-63%); rigour of development, 49% (range 10-81%); clarity of presentation, 55% (range 42-67%); applicability, 14% (range 0-28%) and editorial independence, 28% (range 8-67%). The applicability domain achieved significantly lower quality scores (P = 0.001 versus scope and purpose and P = 0.009 versus rigour of development). AUTHOR'S CONCLUSIONS: Guidelines for the management of pelvic pain associated with endometriosis do not comply with the recommendations for high-quality standards.

Endometriosis↗
PubMed2005

Photo-induced oxidative cross-linking as a method to evaluate the specificity of protein-ligand interactions.

The isolation of protein-binding synthetic molecules from combinatorial libraries or compound collections is now a common practice in chemical biology. An important, but underdeveloped, aspect of characterizing the binding properties of such molecules is their level of binding specificity. This is often evaluated by simply measuring the equilibrium binding affinity of the compound of interest with its target protein and comparing this value with its affinity to one or a few other purified proteins selected at random. These measurements may not reflect accurately the ability of the compound to seek out its target in a complex mixture of proteins such as a cell extract or serum. A more desirable alternative would be to develop solution assays that measure directly the binding of the molecule of interest to both target and competitor proteins in complex solutions. In this report, we evaluate a rapid and efficient photo-triggered cross-linking reaction for assessing binding specificity of synthetic molecules in protein mixtures. Using peptide-protein complexes, we demonstrate that this reaction provides an unbiased view of the peptide-protein contacts present in solution under a given set of conditions and thus is useful for assessing binding specificity. We also discuss the potential application of this chemistry to the related, but more difficult, problem of the identification of protein targets of bioactive molecules.

Amino Acid Sequence↗
PubMed1969

Oral history and the medical librarian.

In recent years there has been an increasing interest in the "oral history," as a technique for creating records or source materials for medical historians. The unique aspects of this technique include the ability to supplement the written record with candid commentary, to create a record or collection of information about an individual or a specific subject area in those instances where a prior record does not exist, and to capture a sample of the personality of the person being interviewed by preservation of the tape recording. There is a lack of unanimity regarding the definition, limits, and goals of oral history. Some individuals insist that the oral historian must in fact be a trained historian and deplore the thought of librarians utilizing this technique. It is the feeling of the author that the interviewer's professional "label" is frequently unimportant, if that individual has prepared himself adequately for the interview. The ideal set of circumstances which combines the oral history memoir with the acquisition and review of the respondent's papers can be handled well by the librarian with the interest and time to conduct such a program. There is general agreement that the time has come when reasonable standards for this technique must be developed if the various programs are to command the interest and respect of those who will use these increasing collections of data. On a more practical level, it is only natural that those who are asked to provide financial support for oral history programs will demand evidence of attention to some sort of standards.

History of Medicine↗
PubMed2006

MCA1 detection of histone H3 serine 10 phosphorylation, a novel biomarker for determination of mitotic indices.

Previously we identified circulating autoantibodies in high titre in the serum of a patient with biopsy-proven discoid lupus erythematosus reactive to serine/threonine phosphoepitopes of the novel mitotic chromosomal autoantigen we named MCA1. MCA1-reactive antibodies have subsequently been demonstrated to bind to the modified histone H3 (H3K9me3S10ph). In this study we utilised Epstein Barr virus (EBV) infection of peripheral blood mononuclear cells collected from this patient to immortalise B-lymphocytes producing MCA1-reactive antibodies. We found MCA1-reactive antibody production by the EBV-immortalised B-lymphocytes unstable in culture. We discuss the possibilities of producing MCA-1-reactive reagents through protein engineering methods (variable or complementarity-determining region grafting) and to modulate antibody affinity through library technologies (such as phage, yeast or ribosome display). Clinical studies have shown that carcinogenesis is most often linked to the development of proliferative abnormalities and proliferative activity has prognostic significance in a variety of human tumours. Here we demonstrate the potential utility of antibodies reactive to H3K9me3S10ph (MCA1), which is only detected on mitotic chromatin, as a marker for cell proliferation in FACS analysis, tissue section staining and in determination of mitotic indices.

Aged↗
PubMed2004

Partnership working in the production of clinical guidelines.

This article describes a working partnership between the Cairns Clinical Librarian Service, Health Care Libraries, University of Oxford and the Emergency Department (ED) at the John Radcliffe Hospital, Oxford. This collaboration resulted in the establishment of a guideline development group in June 2002. The aim of the group is to produce a set of easily accessible, user-orientated, evidence-based guidelines for the use of clinicians in the Emergency Department, within a realistic timescale. The Cairns Clinical Librarian Project was built on previous models of clinical librarianship and incorporated an evaluation of previous programmes. The objective was to provide information to clinicians at the time and point of need. Consultation with the ED from the outset determined the approach adopted by the Cairns team. Clinical guidelines have been shown to improve clinical practice. Pending the introduction of nationally available protocols, we have begun to establish a collection of evidence-based guidelines using web-based architecture in conjunction with the Oxford Radcliffe Hospitals NHS Trust's IT department. A piece of diagnostic analysis was undertaken to ensure a coherent strategy was developed for the guideline project. An 'intermediate approach' was adopted, where existing valid guidelines were modified to fit local circumstances. We strove to be transparent at all times about all aspects of the guideline development process. Formative evaluation has shown how the application of evidence-based health care needs adequate resources and requires people with appropriate knowledge and skills. This article charts the progress of the project and highlights how the partnership between the clinical team and the Clinical Librarian has been deemed to be essential to the success of the project.

Cooperative Behavior↗
PubMed2006

Delayed umbilical cord clamping for reducing anaemia in low birthweight infants: implications for developing countries.

BACKGROUND: Cheap and effective interventions are needed to reduce the risk of infant anaemia in developing countries. Delayed cord clamping (DCC) has been shown to be a simple, safe and cost-free delivery procedure that augments red cell mass in appropriate-for-gestational-age term and preterm infants. It is not known, however, whether DCC is similarly safe and effective in small-for-gestational-age (SGA) infants. We analysed the available evidence to generate a balanced inference on the use of DCC in developing countries. OBJECTIVES: To examine the short- and long-term effects in SGA infants of DCC compared with immediate clamping, and to assess the relationship between time of clamping and the potential postnatal haematological complications of DCC in SGA infants. SEARCH STRATEGY: PubMed (1966 to January 2006), EMBASE (1988 to January 2006) and The Cochrane Library (Issue 1, 2006) were searched. Reference lists of published trials were examined and major journals of perinatal and tropical medicine were hand-searched. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing delayed with immediate cord clamping in infants born between 30 and 42 completed weeks of gestation and which included a proportion of SGA infants. DATA COLLECTION AND ANALYSIS: Three reviewers assessed eligibility and trial quality. MAIN RESULTS: To date, no trials have specifically reported the effects of DCC in SGA infants. Three trials were included, of 190 term and 40 preterm infants, a proportion of whom were SGA. DCC was associated with higher haemoglobin levels in term infants at follow-up [two trials, 127 infants, weighted mean difference (WMD) 9.17 g/L, 95% confidence interval (CI) 5.94-12.40]. In preterm infants, the proportion who required a blood transfusion in the 1st 6 weeks after birth was lower after DCC (one trial, 38 infants, RR 0.56, 95% CI 0.34-0.94). It was not possible to infer from the available data whether SGA infants were at greater risk of adverse effects in the early neonatal period. CONCLUSIONS: DCC in a group that contains both AGA and SGA infants was associated with higher haemoglobin levels at 2-3 months of age in term infants and a reduction in the number of blood transfusions needed in the 1st 4- 6 weeks of life in preterm infants. No reliable conclusions could be drawn about the potential adverse effects of DCC. The paucity of information on DCC in SGA infants justifies further research, especially in developing countries where the baseline risk for polycythaemia-hyperviscosity syndrome is likely to be lower than in industrialised countries.

Anemia↗
PubMed2002

Methadone maintenance therapy versus no opioid replacement therapy for opioid dependence.

BACKGROUND: Methadone maintenance was the first widely used form of opioid replacement therapy developed to treat heroin dependence, and it remains the best-researched treatment for this problem. Despite the widespread use of methadone in maintenance treatment for opioid dependence in many countries, it is a controversial treatment whose effectiveness has been disputed. OBJECTIVES: To evaluate the effects of methadone maintenance treatment (MMT) compared with treatments that did not involve opioid replacement therapy (i.e., detoxification, offer of drug-free rehabilitation, placebo medication, wait-list controls) for opioid dependence. SEARCH STRATEGY: We searched all the following databases up to 2001: Cochrane Drugs and Alcohol Review Group Register, the Cochrane Controlled Trials Register, MEDLINE, EMBASE, Current Contents, Psychlit, CORK [www. state.vt.su/adap/cork], Alcohol and Drug Council of Australia (ADCA) [www.adca.org.au], Australian Drug Foundation (ADF-VIC) [www.adf.org.au], Centre for Education and Information on Drugs and Alcohol (CEIDA) [www.ceida.net.au], Australian Bibliographic Network (ABN), and Library of Congress databases, available NIDA monographs and the College on Problems of Drug Dependence Inc. proceedings, the reference lists of all identified studies and published reviews; authors of identified RCTs were asked about other published or unpublished relevant RCTs. SELECTION CRITERIA: All randomised controlled clinical trials of methadone maintenance therapy compared with either placebo maintenance or other non-pharmacological therapy for the treatment of opioid dependence. DATA COLLECTION AND ANALYSIS: Reviewers evaluated the papers separately and independently, rating methodological quality of concealment of allocation, data were extracted independently for meta-analysis and double-entered. MAIN RESULTS: Six studies met the criteria for inclusion in this review, all were randomised clinical trials, two were double-blind. There were a total number of 954 participants. The method of concealment of allocation was inadequate in one study, not clearly described in four studies, but adequate in a sixth study. Based on the meta-analysis, methadone appeared statistically significantly more effective than non-pharmacological approaches in retaining patient in treatment (3 RCTs, RR=3.05; 95%CI: 1.75-5.35) and in the suppression of heroin use (3 RCTs, RR=0.32; 95%CI: 0.23-0.44), but not statistically in criminal activity (3 RCTs, RR=0.39; 95%CI: 0.12-1.25). REVIEWER'S CONCLUSIONS: Methadone is an effective maintenance therapy intervention for the treatment of heroin dependence as it retains patients in treatment and decreases heroin use better than treatments that do not utilise opioid replacement therapy. It does not show a statistically significant superior effect on criminal activity.

Humans↗
PubMed1995

The impact of IAIMS on the work of information experts. Integrated Advanced Information Management Systems.

Integrated Advanced Information Management Systems (IAIMS) programs differ but have certain characteristics in common. Technological and organizational integration are universal goals. As integration takes place, what happens to those implementing the vision? A survey of 125 staff members, or information experts, involved in information or informatics at an IAIMS-funded institution was conducted during the last year of the implementation phase. The purpose was to measure the impact of IAIMS on the jobs of those in the library and related service units, and the computing, telecommunications, and health informatics divisions. The researchers used newly developed scales measuring levels of integration (knowledge of and involvement with other departments), customer orientation (focus on the user), and informatedness (changes in the nature of work beyond automation of former routines). Ninety-four percent of respondents indicated that their jobs had changed a great deal; the changes were similar regardless of division. To further investigate the impact of IAIMS on librarians in particular, a separate skills survey was conducted. The IAIMS librarians indicated that technology and training skills are especially needed in the new, integrated environment.

Analysis of Variance↗
PubMed2004

Percutaneous central venous catheters versus peripheral cannulae for delivery of parenteral nutrition in neonates.

BACKGROUND: Parenteral nutrition for newborn infants may be delivered via short peripheral cannulae or central venous catheters, which are usually sited percutaneously. The method of delivery may affect nutrient input, and consequently growth and development. Although potentially more difficult to site, percutaneous central venous catheters may be more stable than peripheral cannulae, and need less frequent replacement. These methods may also be associated with different risks of adverse events, including acquired systemic infection and extravasation injury. OBJECTIVES: To review the evidence from randomised controlled trials that, in newborn infants who require parenteral nutrition, infusion via a percutaneous central venous catheter versus a peripheral cannula improves nutrient input, and growth and development, without increasing adverse consequences including systemic infection, or extravasation injuries. SEARCH STRATEGY: We used the standard search strategy of the Cochrane Neonatal Review Group. This included searches of the Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 4, 2003), MEDLINE (1966 - October 2003), EMBASE (1980 - October 2003), conference proceedings, and previous reviews. SELECTION CRITERIA: Randomised controlled trials that compared the effect of delivering parenteral nutrition via percutaneous central venous catheters versus peripheral cannulae in newborn infants. DATA COLLECTION AND ANALYSIS: We extracted the data using the standard methods of the Cochrane Neonatal Review Group, with separate evaluation of trial quality and data extraction by each author, and synthesis of data using relative risk, risk difference and mean difference. MAIN RESULTS: We found three trials eligible for inclusion. These recruited a total of 262 infants and reported a number of different outcomes. One study showed that the use of a percutaneous central venous catheter was associated with a decreased risk of cumulative nutritional deficit during the trial period: Mean difference in the percentage of the prescribed nutritional intake actually received: -7.1% (95% confidence interval -11.2, -3.2). In another trial, infants in the percutaneous central venous catheter group needed significantly fewer catheters/cannulae per infant during the trial period: Mean difference in the number of catheters/cannulae per infant: -3.2 (95% confidence interval -5.13, -1.27). Meta-analysis of data from two trials did not find any evidence of an effect on the incidence of systemic infection: Typical relative risk: 0.90 (95% confidence interval 0.62, 1.32); typical risk difference: -0.04 (95% confidence interval -0.17, 0.10). REVIEWERS' CONCLUSIONS: Data from one small study suggest that the use of percutaneous central venous catheters to deliver parenteral nutrition in newborn infants improves nutrient input. The significance of this in relation to longer-term growth and developmental outcomes is unclear. Another study suggested that the use of percutaneous central venous catheters rather than peripheral cannulae decreases the number of catheters/cannulae needed to deliver the nutrition. We have not found any evidence that percutaneous central venous catheter use increases the risk of adverse events, particularly systemic infection.

Catheterization, Central Venous↗
PubMed2006

The Macrostomum lignano EST database as a molecular resource for studying platyhelminth development and phylogeny.

We report the development of an Expressed Sequence Tag (EST) resource for the flatworm Macrostomum lignano. This taxon is of interest due to its basal placement within the flatworms. As such, it provides a useful comparative model for understanding the development of neural and sensory organization. It was anticipated on the basis of previous studies [e.g., Sánchez-Alvarado et al., Development, 129:5659-5665, (2002)] that a wide range of developmental markers would be expressed in later-stage macrostomids, and this proved to be the case, permitting recovery of a range of gene sequences important in development. To this end, an adult Macrostomum cDNA library was generated and 7,680 Macrostomum ESTs were sequenced from the 5' end. In addition, 1,536 of these aforementioned sequences were sequenced from the 3' end. Of the roughly 5,416 non-redundant sequences identified, 68% are similar to previously reported genes of known function. In addition, nearly 100 specific clones were obtained with potential neural and sensory function. From these data, an annotated searchable database of the Macrostomum EST collection has been made available on the web. A major objective was to obtain genes that would allow reconstruction of embryogenesis, and in particular neurogenesis, in a basal platyhelminth. The sequences recovered will serve as probes with which the origin and morphogenesis of lineages and tissues can be followed. To this end, we demonstrate a protocol for combined immunohistochemistry and in situ hybridization labeling in juvenile Macrostomum, employing homologs of lin11/lim1 and six3/optix. Expression of these genes is shown in the context of the neuropile/muscle system.

Animals↗
PubMed2001

A CE-MALDI interface based on the use of prestructured sample supports.

We have developed an off-line coupling of capillary electrophoresis (CE) to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS) based on CE fraction collection onto prestructured MALDI sample supports. Analyte carryover and detection sensitivity were investigated using a standard peptide mixture. Low femtomole amounts were detected, and no noticeable carryover was discovered. The performance of the method was evaluated with a mixture of tryptic digests of proteins from a human fetal brain cDNA expression library. The total number of identified peptides was increased from 47 to 211 when the CE-MALDI interface was used compared to direct MALDI-MS analysis. Sequence coverage with CE-MALDI was in the 25-60% range for the different proteins, corresponding to an increase of 1.3-4.9 times relative to that obtained with MALDI-MS of the crude mixture. Fractionation of sample components also facilitated protein identification by MALDI postsource decay analysis. Our initial results suggest this CE-MALDI interface can be used for the analysis of complex peptide mixtures isolated from biological tissues.

Brain Chemistry↗
PubMedDate not supplied

Development of a mass-directed preparative supercritical fluid chromatography purification system.

In this paper, we report the development of a mass-directed supercritical fluid chromatography (SFC) purification system. We have addressed issues on software compatibility, the interface between the preparative SFC and the mass spectrometer, and fraction collection. Good peak shape and signal were achieved in the mass spectrometry (MS) trace, allowing accurate peak detection and reliable fraction collection. Simple modifications on a commercially available fraction collector enabled fractionation at atmospheric pressure with high recovery. The SFC/MS purification system has been used in support of high-throughput library purification and has been proven to be a valuable tool in complementing our reversed-phase high-performance liquid chromatograph (RP-HPLC/MS)-based technology platform.

Chromatography, Supercritical Fluid↗