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PubMed2005

Perspectives for systematic in vitro antibody generation.

After the completion and refinement of the human genome, the characterization of individual gene products in respect of their functions, their modifications, their cellular localization and regulation in both space and time has generated an increased demand for antibodies for their analysis. Taking into account that the human genome contains approximately 25,000 genes, and that their products are found in different splice variants and produce proteins with post-translational modifications, it can be estimated that at least 100,000 different protein products have to be investigated to gain a complete picture of what's going on in the proteome of a cell. Antibodies are preferred tools helping with the characterization and detection of proteins as well as with elucidating their individual functions. The generation of antibodies to all available human protein products by immunization and/or the hybridoma technology is not only logistically and financially enduring, but may prove to be a difficult task, as quite a number of interesting targets may evade the immune response of experimental animals, for example, allosteric variants dependent on fragile interactions to cofactors, highly conserved antigens etc. For this reason, alternative methods for the generation of antibodies have to supplement these approaches. In vitro methods for antibody generation are seen to offer this capability. In addition, they may provide a cost effective and large scale production alternative for detection reagents for the research community in their own right. Among in vitro techniques, phage display has been evolved as the most efficient option for tackling this problem and approaches optimised for automation are emerging. Maximum benefit for proteomic research could be generated by judicious and preferably international coordination of the ongoing efforts to combine the strengths of the well established animal based approaches and the novel opportunities offered by in vitro methods.

Antibodies↗
PubMed2006

Prevalence of congenital heart disease in schoolchildren of sub-Saharan Africa, Mozambique.

A survey conducted by cardiologists in Mozambique provided an opportunity for assessing the prevalence of congenital heart disease in public school children in Maputo. After randomization, a total of 2170 children were examined. Five children had clinical and echographic evidence of congenital heart disease, corresponding to a prevalence of 2.3 in 1000 (95% confidence interval: 1.0 to 5.4), 80% newly discovered during the survey. These findings confirm that congenital heart disease is at least as common in this South African black community as in Caucasians. Early detection of these cardiac diseases in children is important in order to prevent serious complications and to institute endocarditis prevention.

Adolescent↗
PubMed2004

13CO2 pulse labelling of plants in tandem with stable isotope probing: methodological considerations for examining microbial function in the rhizosphere.

Recently developed 13CO2 pulse labelling and stable isotope probing (SIP) methods offer the potential to track 13C-labelled plant photosynthate into phylogenetic groups of microbial taxa in the rhizosphere, permitting an examination of the link between soil microbial diversity and carbon flow in situ. We tested the feasibility of this approach to detect functional differences in microbial communities utilising recently fixed plant photosynthate in moisture perturbed grassland turfs. Specifically, we addressed two questions: (1) How does moisture perturbation (three treatments; continual wetting, drying, and drying followed by rewetting) affect the assimilation of 13C-labelled exudates carbon into the soil microbial community?; (2) Can 13C deposited in soil from pulse-labelled plants be used to identify microbes utilising plant exudates using SIP methodologies? Net CO2 fluxes showed that prior to 13CO2 pulse labelling, all treatments were photosynthetically active, but differences were observed in night time respiration, indicating moisture treatments had impacted on net CO2 efflux. Measurements of pulse-derived 13C incorporated into soil RNA over 2 months showed that there was only evidence of 13C enrichment in the continuously wetted treatments. However, isotopic values represented only a 0.1-0.2 13C at.% increase over natural abundance levels and were found to be insufficient for the application of RNA-SIP. These findings reveal that in this experimental system, the microbial uptake of labelled carbon from plant exudates is low, and further optimisation of methodologies may be required for application of SIP to natural plant-soil systems where 13C tracer dilution is a consideration.

Carbon Dioxide↗
PubMed2006

Detection of methanogenic Archaea in peat: comparison of PCR primers targeting the mcrA gene.

Methanogens (domain Archaea) have a unique role in the carbon cycle as producers of the greenhouse gas methane (CH(4)). Methyl-coenzyme M reductase (MCR) is a vital enzyme in CH(4) production, and the mcrA gene coding for a subunit of MCR has been employed as a specific marker for the detection and differentiation of methanogen communities. A critical step in assessing environmental mcrA diversity is the selection of PCR primers. The objective of this study was to compare the diversity coverage of three published mcrA primer sets MCR, ME and ML (also known as MCR and Luton-mcrA) and their ability to discern methanogen communities in a drained peatland. The primers were applied to DNA extracts from unfertilised and ash-fertilised peat from two different depths. Amplified mcrA communities were cloned and sequenced, and the sequences were divided into operational taxonomic units (OTUs) by restriction fragment length polymorphism (RFLP) and sequence analysis. All primers recovered characteristic OTUs associated with the peat depths and treatments and confirmed a previous observation of low methanogen diversity. The minor differences in OTU ranges of the primers did not greatly affect the observed community composition. However, as the proportions of several OTUs varied strongly, the primers provided different quantitative representations of mcrA communities. We concluded that the ML and MCR primers had better amplification ranges than the ME set, but the use of MCR with peat samples was problematic due to poor amplification. Consequently, the ML primers were best suited for mcrA analysis of peatland methanogen communities.

Amino Acid Sequence↗
PubMed2005

Smoking cessation intervention in a large randomised population-based study. The Inter99 study.

BACKGROUND: Several large and well-conducted community interventions have failed to detect an effect on prevalence of smoking. METHODS: Two thousand four hundred eight daily smokers in all motivational stages were actively recruited and included in a randomised population-based intervention study in Copenhagen, Denmark. All smokers completed a questionnaire and underwent a health examination and a lifestyle consultation. Daily smokers in the high intensity intervention group were offered assistance to quit in smoking cessation groups. RESULTS: The validated abstinence rate at 1-year follow-up was 16.3% in the high intensity group and 12.7% in the low intensity group compared with a self-reported abstinence rate of 7.3% in the background population. The adjusted odds ratio of abstinence in the high intervention group was significantly higher, OR = 2.2 (1.6-3.0) than in the background population, also in the 'intention-to-treat' analyses, OR = 1.5 (1.1-2.0). Higher socioeconomic status, higher age at onset of daily smoking, and a higher wish to quit were predictors of success. CONCLUSION: In a population-based setting, using active recruitment and offering assistance to quit, it was possible to include many smokers and to achieve a significantly higher validated abstinence in the high intensity intervention than in the background population, even when using 'intention-to-treat' analyses.

Adult↗
PubMed1997

Correlates of unrecognized acute myocardial infarction detected via perfusion imaging.

Research using the electrocardiogram (ECG) indicates that about 1/3 of acute myocardial infarctions (AMIs) are unrecognized. To date, no studies of unrecognized AMIs have employed perfusion imaging, although it is more sensitive than the ECG and provides more information about infarct characteristics, such as size and location. In this study, 82 of 258 consecutive patients (31.8%) undergoing exercise testing with technetium-99m sestamibi perfusion imaging had fixed, nonartifactual perfusion defects, suggesting AMI. These patients were interviewed regarding their recognition of AMI; 27 patients (32.9%) had unrecognized AMI. Unrecognized AMI was significantly associated with (1) smaller infarcts, (2) infarcts not in the apical or septal regions, (3) diabetes mellitus, (4) lack of angina, (5) a negative family history for cardiac disease, and (6) being African-American. Many of these variables were significantly intercorrelated, and in multivariate analysis, unrecognized AMI remained significantly predicted by a smaller infarct and lack of angina. This study suggests that the incidence of unrecognized AMI detected via perfusion imaging on a clinic population is similar to that detected via electrocardiographic studies on community samples. This study also replicates prior findings of the medical history and demographic correlates of unrecognized AMI, and indicates that infarct size and location are also associated with unrecognized AMI.

Aged↗
PubMed1998

The future: screening and effective intervention.

Despite an increased understanding of risk factors for cardiovascular disease and the development of new programs, procedures, and medications to reduce risk, effective large-scale primary and secondary prevention have remained difficult to achieve. Large-scale, primarily educational, community-based programs aimed at detecting and reducing risk can be effective, particularly when they are aimed at specific populations at increased risk. However, the long-term benefits of these programs have been modest. Recent studies have demonstrated that lipid-lowering therapy is a critical adjunct to dietary and lifestyle changes. The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors in particular are highly effective in reducing cardiovascular risk in both primary and secondary prevention programs that combine lifestyle modification and aggressive medical therapy. The ability of these drugs to produce rapid improvements in endothelial function and increase coronary perfusion also supports their use in the medical rather than surgical management of at least some patients with cardiovascular disease who are candidates for percutaneous transluminal coronary angioplasty.

Angioplasty, Balloon, Coronary↗
PubMed2002

Analysis on residues of estrogens, gestagens and androgens in kidney fat and meat with gas chromatography-tandem mass spectrometry.

The use of estrogens, gestagens and androgens (EGAs) in animal fattening is prohibited in the European Community. Based on the general detection capabilities of Belgian laboratories, National Minimum Required Performance Limits (National MRPLs) for a number of EGAs have been imposed by the inspection services. Selective hyphenated techniques, e.g. GC-MS and GC-MS2, with high detection capability are needed. Beta-trenbolone, which is meant to be a "problem" molecule for GC-MS, can be detected at the 2 microg/kg level using GC-MS2. Based on the National MRPLs in different matrices, our laboratory has divided the EGAs into a class system. In this set-up, analysis of EGAs in kidney fat and meat is discussed.

Adipose Tissue↗
PubMed2000

Biotic indices based on soil nematode communities for assessing soil quality in terrestrial ecosystems.

Nematode communities offer several advantages for assessing the quality of soils. Diversity and maturity indices as well as trophic structure defining parameters based on the composition of soil nematode communities have been used to detect various kinds of man-induced disturbances and monitor ecological recovery. In the present paper we examined the response of these indices in herbaceous Mediterranean ecosystems. Incipient soils, which were developed on the top of waste dumping sites were compared with agricultural soils under fallow. Grasslands and old-fields were considered as reference systems. Either diversity or maturity indices were shown to be very sensitive to changes in soil system caused by waste dumping. However, they were less useful for the categorisation of perturbation in agroecosystems. Trophic composition related parameters were more meaningful for this aim. Plant parasite index (PPI) may be a good generic indicator of recovery processes after perturbation in these annual herbaceous ecosystems.

Animals↗
PubMed2005

Cumulative incidence of childhood autism: a total population study of better accuracy and precision.

Most studies on the frequency of autism have had methodological problems. Most notable of these have been differences in diagnostic criteria between studies, degree of cases overlooked by the initial screening, and type of measurement. This study aimed to replicate the first report on childhood autism to address cumulative incidence as well as prevalence, as defined in the International Statistical Classification of Diseases and Related Health Problems, 10th revision (ICD-10) Diagnostic Criteria for Research. Here, the same methodological accuracy (exactness of a measurement to the true value) as the first study was used, but population size was four times larger to achieve greater precision (reduction of random error). A community-oriented system of early detection and early intervention for developmental disorders was established in the northern part of Yokohama, Japan. The city's routine health checkup for 18-month-old children served as the initial mass screening, and all facilities that provided child care services aimed to detect all cases of childhood autism and refer them to the Yokohama Rehabilitation Center. Cumulative incidence up to age 5 years was calculated for childhood autism among a birth cohort from four successive years (1988 to 1991). Cumulative incidence of childhood autism was 27.2 per 10000. Cumulative incidences by sex were 38.4 per 10000 in males, and 15.5 per 10000 in females. The male:female ratio was 2.5:1. The proportions of children with high-functioning autism who had Binet IQs of 70 and over and those with Binet IQs of 85 and over were 25.3% and 13.7% respectively. Data on cumulative incidence of childhood autism derived from this study are the first to be drawn from an accurate, as well as precise, screening methodology.

Autistic Disorder↗
PubMedDate not supplied

The control of Chagas' disease in Mambaí, Brazil: the initial phases.

The control of Chagas' disease depends on breaking contact between Man and vector organisms living in the house fabric; the primary method utilized is residual insecticide spraying of houses. Ministries of health in afflicted countries have elaborated standard procedures for such combat programs. The first two phases of such a program in Mambaí, Goiaś, Brazil are described. In the third phase of vigilance, community participation is necessary to detect residual organism populations in houses. A house improvement scheme applied to houses with persistent infection, is recommended as being most practical at this time.

Brazil↗
PubMed1990

A hand-held OKP chart for the screening of glaucoma: preliminary evaluation.

Oculo-kinetic perimetry (OKP) uses controlled movements of the patient's eye to position a static test stimulus in the visual field. The test chart consists of a white tangent screen with 23 numbered fixation points located eccentrically at strategic points in relation to a central black spot, which is the test stimulus. As the patient looks at each number in turn, the spot stimulates the retinal areas that are most likely to show early glaucomatous damage. The result is positive if one or more numbers make the spot disappear. Ninety-eight eyes of 54 patients attending a hospital glaucoma clinic were tested with a prototype OKP glaucoma screener and by conventional perimetry. Visual fields plotted conventionally were independently categorised as normal, equivocal and glaucomatous by the perimetrist and the ophthalmologist and these results were further categorised as normal, equivocal or glaucomatous according to whether they were considered abnormal by none, one or both of the observers respectively. The OKP test was positive in (i) 93% of 27 eyes with glaucomatous defects, (ii) 69% of 32 eyes with equivocal loss, (iii) 41% of 39 eyes without previously recognised field loss but having ocular hypertension, a suspicious disc, or contralateral glaucoma, (iv) 9% of 116 eyes from 60 age matched persons accompanying the patients to hospital. If the results were considered in terms of patients not eyes, the OKP test was positive in 85% individuals with glaucomatous field loss in their worse eye and in 12% of the controls. A hand-held OKP chart testing a small set of points and using a constantly exposed stimulus could be a useful tool for the detection of glaucoma in the community.

Evaluation Studies as Topic↗
PubMed1998

Bladder tumours detected on screening: results at 7 years.

OBJECTIVES: To report the outcome at a minimum follow-up of 7 years of a cohort of tumours that were asymptomatic at presentation and detected in a previous community-based screening programme (with reduced mortality and progression at 3 years of follow-up). PATIENTS AND METHODS: In the original screening, 2356 men aged > or = 60 years had a dipstick test for microscopic haematuria; 474 (20%) had positive results and of these, 317 agreed to further investigation. Twenty-one were found to have bladder tumours; this was reduced to 17 after pathological review. No tumours were muscle-invasive at diagnosis. The outcome of this cohort was evaluated at 7 years of follow-up. RESULTS: The patients were grouped on the basis of their original histology. In group A, four patients had no tumour on histological review and none progressed in grade or stage. In group B, eight patients reported to have well differentiated superficial tumours (i.e. G1pTa) had no recorded deaths from cancer or progression to muscle-invasive disease. In group C, of nine patients with tumours having a potentially worse prognosis (G1-2, > or = pT1, or carcinoma in situ, three died from bladder cancer, two progressed to muscle-invasive disease (one of whom died from unrelated causes) and two further patients died from unrelated causes. CONCLUSION: This cohort of patients represents a unique group, in that all had superficial disease at the time of diagnosis. The outcome at 3 years was excellent, but in the subsequent 4 years many patients had progression of their cancer. All patients who developed invasive disease were in group C; as their disease was identified at a superficial stage it may have been amenable to aggressive early management not usually associated with such grade and stage.

Aged↗
PubMed1998

Persistent parasitemia after acute babesiosis.

BACKGROUND: Babesiosis, a zoonosis caused by the protozoan Babesia microti, is usually not treated when the symptoms are mild, because the parasitemia appears to be transient. However, the microscopical methods used to diagnose this infection are insensitive, and few infected people have been followed longitudinally. We compared the duration of parasitemia in people who had received specific antibabesial therapy with that in silently infected people who had not been treated. METHODS: Forty-six babesia-infected subjects were identified from 1991 through 1996 in a prospective, community-based study designed to detect episodes of illness and of seroconversion among the residents of southeastern Connecticut and Block Island, Rhode Island. Subjects with acute babesial illness were monitored every 3 months for up to 27 months by means of thin blood smears, Bab. microti polymerase-chain-reaction assays, serologic tests, and questionnaires. RESULTS: Babesial DNA persisted in the blood for a mean of 82 days in 24 infected subjects without specific symptoms who received no specific therapy. Babesial DNA persisted for 16 days in 22 acutely ill subjects who received clindamycin and quinine therapy (P=0.03), of whom 9 had side effects from the treatment. Among the subjects who did not receive specific therapy, symptoms of babesiosis persisted for a mean of 114 days in five subjects with babesial DNA present for 3 or more months and for only 15 days in seven others in whom the DNA was detectable for less than 3 months (P<0.05); one subject had recrudescent disease after two years. CONCLUSIONS: When left untreated, silent babesial infection may persist for months or even years. Although treatment with clindamycin and quinine reduces the duration of parasitemia, infection may still persist and recrudesce and side effects are common. Improved treatments are needed.

Animals↗
PubMed2006

Help-seeking rates for intimate partner violence (IPV) among Canadian immigrant women.

We examined rates of help seeking for intimate partner violence (IPV) among recent (0-9 years in Canada) and non-recent (10+ years in Canada) immigrant women. Data from a national, cross-sectional, telephone survey were used. Help-seeking variables included disclosure of IPV, reporting IPV to police, use of social services subsequent to IPV, and barriers to social service use. Recent immigrant women, compared with non-recent immigrant women, were significantly more likely to report IPV to police and less likely to use social services. Findings have important implications for prevention and detection of IPV in immigrant communities and in future research.

Adult↗
PubMed2003

Volatile substance and other drug abuse inhalation in Spain.

OBJECTIVE: Inhalant misuse is the intentional inhalation of volatile substances in order to obtain euphoric, disinhibiting, and exciting effects. Solvents, glues, adhesives, paints, varnishes, paint removers, dry cleaning agents, spray paints, nail polish removers, typewriter correction fluids, and aerosol propellants are common sources of volatile substance abuse. In recent years the abuse of inhalant substances, not only among those who abuse other drugs but also in teenagers and younger children, has been reported. We reviewed retrospectively the cases of inhalant misuse reported to the Spanish Poison Control Center. METHODS: Human intoxications from abuse of inhalant substances registered by our service from 1991 to 2000 were studied. Data analyzed were relative to age, gender, signs and symptoms, drug dependence antecedents, and severity of symptoms of the patients. The type of product and composition were also investigated. RESULTS: During the study period 109 cases of patients aged from 8 to 50 years were collected. A percentage of 36.6% was less or equal to 20 years old. Seventy percent corresponded to males. Of the patients, 11% presented dependence antecedents to other abuse drugs and 72.5% were symptomatic. In the symptomatic exposures clinical features affected the following systems: CNS (62.8%), gastrointestinal (8.1%), cardiovascular (8.1%), respiratory (2.9%), peripheral nervous system (1.1%), renal (1.1%), haematological (1.1%), hepatic (1.7%), and other (13.1%). The commercial products more frequently inhaled were solvents (34.9%) and glues/adhesives (22.9%). We noted the use of medicines with ethyl chloride-local anaesthetic (8.3%), three cases with aerosol bronchodilator (with fluorocarbons as propellants), and one case of xylazine inhalation. The composition most often involved was aromatic hydrocarbons (46.9%), halogenated hydrocarbons (16.5%), aliphatic hydrocarbons (11.4%), ketones (10.1%), local anaesthetic (ethyl chloride) (8.4%), ethers (2.5%), nitrous oxides (2.5%), and aliphatic nitrites (1.7%). The calls received were 59.6% from health care units and 22% from general public. Only 14% of cases were at home and 48% had moderate to severe clinical effects. Acute intoxications occurred in 82% of cases. CONCLUSIONS: Inhalation of volatile substance as abuse drugs has been detected in different age groups, including very young people. Although the principal source was industrial products, the use of drugs such as local anaesthetics and aerosol broncodilators was also detected. Based on epidemiological studies in the Spanish population (essentially adolescents and childhood) together with the ability of a Poison Center to detect sentinel-events, the community and authorities should develop strategies to prevent these exposures and the later use of other substances of abuse. In fact, recently a Law on Drug Dependences and Other Addictive Alterations has been approved in Madrid in order to take precautionary measures.

Administration, Inhalation↗
PubMed2003

Natural and nosocomial infection in a patient with West Nile encephalitis and extrapyramidal movement disorders.

Since its first recognition in North America in 1999, West Nile virus (WNV) has spread rapidly across the continent, but in many communities, rapid diagnostic tests for detection of WNV infection are not fully available. We describe a patient with extrapyramidal movement disorders and changes in the basal ganglia noted on magnetic resonance images that are characteristic of other flavivirus encephalitides and may help in the recognition of patients with West Nile encephalitis. Detailed molecular analysis suggested that, although our patient received a blood transfusion infected with WNV, the virus that caused his initial infection and encephalitis was probably acquired naturally from a mosquito.

Animals↗
PubMed1998

Tay-Sachs disease carrier screening: a model for prevention of genetic disease.

Tay-Sachs disease (TSD) is an autosomal-recessive, progressive, and ultimately fatal neurodegenerative disorder. Within the last 30 years, the discovery of the enzymatic basis of the disease, namely deficiency of the enzyme hexosaminidase A, made possible both enzymatic diagnosis of TSD and heterozygote identification. In the last decade, the cloning of the HEXA gene and the identification of more than 80 associated TSD-causing mutations has permitted molecular diagnosis in many instances. TSD was the first genetic condition for which community-based screening for carrier detection was implemented. As such, the TSD experience can be viewed as a prototypic effort for public education, carrier testing, and reproductive counseling for avoiding fatal childhood disease. More importantly, the outcome of TSD screening over the last 28 years offers convincing evidence that such an effort can dramatically reduce incidence of the disease.

Alleles↗