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At least 883 records · Page 49Linked to original sources
PubMed2001

[Simplifying the sampling method for evaluating the larval density of Aedes aegypti in São Paulo State, Brazil].

OBJECTIVE: To suggest changes in the sampling procedures for estimating the larval density of Aedes aegypti in the state of São Paulo, Brazil, by assessing the efficacy and level of complexity of the current procedures. METHODS: The "Superintendência de Controle de Endemias do Estado de São Paulo" - SUCEN's (Superintendence for Endemic Control of the State of São Paulo) sampling procedures for estimating the density of Aedes aegypti larvae was assessed. The Breteau index with one-stage cluster sampling was applied. An average of 35 blocks, 14 buildings per block, of a total of 510 buildings per sample, were assessed. One hundred and eleven samples were obtained in 6 municipalities of São José do Rio Preto. RESULTS: The Breteau index estimates higher than 3 had coefficients of variation smaller than 30% in 71% of the samples. The design effect was 1.19 and 1.79 for indexes lower and higher than 5, respectively. The estimates were obtained in less than 3 days, requiring a daily average of 5.5 men per sampling. CONCLUSION: This sampling design revealed to be a simplified one: fast, economic and easily managed. The operational easiness was due to the use of the one-stage cluster sampling, when there was not need to prepare lists of addresses and then sort and locate the buildings. Though the design effect shows a slight loss of the estimates' accuracy with the use of one-stage cluster sampling, this was proven to be under acceptable range. Simplifying changes in the process of determining the sample size and assessing sampling errors are proposed.

Aedes↗
PubMed2003

[Master sample and geoprocessing: technologies for household surveys].

OBJECTIVE: To reduce cost and time associated with household sampling process and to assess the feasibility of shared use of address data file of census enumeration areas in several epidemiological surveys using updated information from the National Survey of Households (PNAD). METHODS: Address data file comprising 72 census enumeration areas was kept as primary sampling units for the city of S o Paulo. During the period 1995-2000, three distinct household samples were drawn using the two-stage cluster sampling procedure. Geographic Information System (GIS) technology allowed delimiting boundaries, blocks and streets for any primary sampling unit and printing updated maps for selected sub-samples. RESULTS: Twenty-five thousand dwellings made up the permanent address data file of the master sample. A cheaper and quicker selection of each sample, plus gathering information on demographic and topographical profiles of census enumeration areas were the main contribution of the study results. CONCLUSIONS: The master sample concept, integrated with GIS technology, is an advantageous alternative sampling design for household surveys in urban areas. Using the list of addresses from the PNAD updated yearly, although limiting its application to the most populated Brazilian cities, avoids the need of creating an independent sampling procedure for each individual survey carried out in the period between demographic censuses, and it is an important contribution for planning sampling surveys in public health.

Censuses↗
PubMed1991

Epidemiologic studies utilizing surveys: accounting for the sampling design.

BACKGROUND: Since large-scale health surveys usually have complicated sampling schemes, there is often a question as to whether the sampling design must be considered in the analysis of the data. A recent disagreement concerning the analysis of a body iron stores-cancer association found in the first National Health and Nutrition Examination Survey and its follow-up is used to highlight the issues. METHODS: We explain and illustrate the importance of two aspects of the sampling design: clustering and weighting of observations. The body iron stores-cancer data are reanalyzed by utilizing or ignoring various aspects of the sampling design. Simple formulas are given to describe how using the sampling design of a survey in the analysis will affect the conclusions of that analysis. RESULTS: The different analyses of the body iron stores-cancer data lead to very different conclusions. Application of the simple formulas suggests that utilization of the sample clustering in the analysis is appropriate, but that a standard utilization of the sample weights leads to an uninformative analysis. The recommended analysis incorporates the sampling weights in a nonstandard way and the sample clustering in the standard way. CONCLUSIONS: Which particular aspects of the sampling design to use in the analysis of complex survey data and how to use them depend on certain features of the design. We give some guidelines for when to use the sample clustering and sample weights in the analysis.

Epidemiologic Methods↗
PubMed2000

The design of multi-stage tuberculin surveys: some suggestions for sampling.

SETTING: Tuberculin surveys of children to estimate national or regional infection prevalences are commonly designed as multi-stage surveys. These surveys require strategies for the efficient sampling of sub-units at each stage. OBJECTIVES: To develop guidelines for sampling in tuberculin surveys. DESIGN: Sampling theory was used to develop a simple and efficient sampling strategy for planning and analysing tuberculin surveys. The issue of sample sizes is considered. RESULTS: Formulae for the calculation of infection prevalences and their confidence intervals are developed. Sample sizes are discussed. CONCLUSION: We recommend that districts be sampled using sampling proportional to size, in which larger units have a larger probability of being included in the sample. Schools are sampled next using simple random sampling, where each school within a district has the same probability of being included in the sample. In each school all eligible children are Mantoux tested. The number of children tested per district should be approximately constant. Increasing the number of selected districts is usually more efficient for increasing the precision of the estimate than increasing the number of children per district beyond several hundred to a few thousand.

Bias↗
PubMed1998

Deleted HTLV-1 provirus in cord-blood samples of babies born to HTLV-1-carrier mothers.

We screened 596 cord-blood samples by nested short PCRs for the gag and pX regions of HTLV-1 which are capable of detecting a single copy. The samples were derived from 449 and 147 babies born to seropositive and seronegative mothers respectively. Of these, 20 samples were positive in at least one of the PCRs: 9 (45%) were positive in both PCRs, but 10 and 1 samples in either the pX or the gag PCR respectively. These samples were tested further in nested long PCRs directed for gag-pX, gag-pol and pol-pX regions capable of detecting 8, 1 and 2 copies respectively. Of 9 dually positive samples, 7 (77%) showed the predicted 6.2-kbp band in the gag-pX PCR; only 2 of them had the predicted band alone; 7 samples had discrete bands shorter than the predicted size. In the gag-pol PCR, all 9 samples showed the predicted 2.2-kbp band alone. In the pol-pX PCR, 819 samples showed the predicted 4.2-kbp band, including one with an additional 2.1-kbp band, and the last a 1.0-kbp band alone. Thus, all of the dually positive samples had proviruses harboring gag, pol and pX priming sites. In contrast, none of the 11 singly positive samples showed the predicted band in the gag-pX PCR: 5 had no visible band, and the other 6 had shorter bands only. None of these 11 samples showed any positive signal in either gag-pol or pol-pX PCR. Our results suggest that HTLV-1 proviruses in the cord blood are frequently defective.

Base Sequence↗
PubMed2004

Comparison of hybrid capture 2 with in situ hybridization for the detection of high-risk human papillomavirus in liquid-based cervical samples.

BACKGROUND: The performance of two commercially available detection systems for high-risk HPV (hrHPV), Hybrid Capture 2 (HC2) and in situ hybridization (ISH), were compared on cervical scrapings. METHODS: Using general primer (GP)-mediated GP5+/6+-polymerase chain reaction (PCR)-enzyme immunoassay and reverse line blot genotyping, 76 liquid-based cervical samples were identified with > or = 1 of the 12 hrHPV types present in the probes of the HC2 and ISH assays. The positivity rate of the assays and the HC2 viral load were determined and related to cytologic findings (n = 76 samples) and histologic findings (n = 43 samples). RESULTS: Overall, HC2 scored significantly more samples positive compared with ISH (P < 0.01). Seventy-four of 76 samples (97%) were positive according to HC2. Forty-six of 76 samples (61%) were positive according to ISH, including 80% and 70% of samples that were classified cytologically as moderate dysplasia and severe dysplasia, respectively. All women with underlying cervical intraepithelial neoplasia (CIN) lesions and 67% of women without CIN had positive HC2 samples. ISH scored 33%, 66%, 88%, and 73% of samples positive of women with no CIN, Grade 1 CIN (CIN 1), CIN 2, and CIN 3, respectively. The HC2 viral load was significantly higher in women who had a cytologic diagnosis of dysplasia (P < 0.01) and in women who had an underlying diagnosis of CIN (P < 0.01) compared with women who had neither. In addition, the viral load was significantly higher in ISH positive samples compared with ISH negative samples (P < 0.01). CONCLUSIONS: An increased HC2 viral load was associated with an increased chance of underlying high-grade CIN disease in women who tested hrHPV GP5+/6+-PCR positive. Moreover, although positive ISH results were associated with an increased overall viral load in the sample, the analytic sensitivity of ISH was too low to detect all women with prevalent high-grade CIN.

Adult↗
PubMed2006

New immunochemical fecal occult blood test with two-consecutive stool sample testing is a cost-effective approach for colon cancer screening: results of a prospective multicenter study in Chinese patients.

The purpose of the study is to evaluate a new immunochemical fecal occult blood test method (Hemosure IFOBT), and compare it to the Guaiac-based chemical method (CFOBT) for colorectal cancer detection. A hypothetical sequential method (SFOBT), in which IFOBT was used only as a confirmatory test for CFOBT, was also evaluated. A total of 324 patients were recruited from 5 major hospitals in Beijing, China. For each patient, 3 consecutive stool samples were collected for simultaneous CFOBT and IFOBT tests, followed by colonoscopic examination. We compared the sensitivity and specificity of the 3 methods (CFOBT, IFOBT and SFOBT) in two settings, with the first 2 consecutive samples versus all 3 samples. Although the sensitivity for the detection of cancer and large (>20 mm) or multiple adenoma was similar for all 3 methods in the three-sample setting, in the two-sample setting IFOBT had higher sensitivity than SFOBT for detecting cancer (87.8% vs. 75.5%, respectively, p < 0.05) and large (>20 mm) or multiple adenomas (65.4% vs. 42.3%, respectively, p < 0.05). The IFOBT also had a higher specificity than the CFOBT (89.2% vs. 75.5%, respectively, p < 0.01) in "normal" individuals defined by colonoscopy in the three-sample setting. Comparing two-sample setting to the three-sample setting, both CFOBT and SFOBT showed significant loss of sensitivity for the detection of cancer as well as adenoma, whereas the sensitivity for IFOBT did not change significantly. Overall, IFOBT with two-sample testing showed compatible sensitivity and specificity to the three-sample testing, and had a lower relative cost per cancer detected than the three-sample testing. In conclusion, the new Hemosure IFOBT with two consecutive stool samples appears to be the most cost-effective approach for colon cancer screening.

Adenoma↗
PubMed1998

Development of a limited sampling approach in pharmacokinetic studies: experience with the antiepilepsy drug tiagabine.

A sparse sampling method is proposed to assess pharmacokinetic parameters after a single dose of the antiepilepsy drug tiagabine. Pharmacokinetic parameters obtained from two different pharmacokinetic studies were compared using sparse sampling (7 blood samples) with extensive sampling (15 to 16 blood samples). The results indicated that sparse blood samples taken at appropriate times can be used to estimate pharmacokinetic parameters as accurately as extensive blood samples. In addition, a limited sampling model (LSM) was developed using samples from 10 subjects at two time points (6 and 8 hours). The model was validated in 40 subjects and provided good population mean estimates of area under the concentration-time curve (AUC) and maximum concentration (Cmax). The sparse sampling method described here can be used to assess pharmacokinetic parameters in drug development provided a prior knowledge of the pharmacokinetics of a drug has been obtained from extensive sampling. Further, the LSM described here may be useful in estimating AUC and Cmax of tiagabine using two samples in clinical settings. The LSM approach described here can also be used to estimate AUC and Cmax of a drug in preclinical toxicokinetic studies without detailed pharmacokinetic studies.

Adult↗
PubMed2004

Structural and antigenic preservation of plant samples by microwave-enhanced fixation, using dedicated hardware, minimizing heat-related effects.

We explored the use of microwave technology in fixation with the objective of achieving quicker fixation regimes, lower concentrations of toxic and volatile reagents, and enhanced antigen detection. We used a modified domestic microwave oven (900 W) and a low-power (5 W) microwave bench. The work was done on plant materials. The oven was supplemented with a cooling device, a stirring system, and a record of the sample temperature and the time of effective irradiation. The sample, immersed in a fixative solution of 1% paraformaldehyde (PFA) in PBS, was irradiated for only 10 minutes. The sample temperature did not exceed 37 degrees C. In these mild conditions, the quality of the (ultra)structural preservation of the samples, morphometrically assessed, was at the same level as obtained with the same fixative, using conventional methods. On the contrary, samples fixed in the same conditions without irradiation showed a poor structural preservation. The antigenic preservation of the irradiated samples was excellent, since the labeling levels of two nucleolar proteins, detected by immunogold, were three times higher than in conventionally fixed samples. In the so-called microwave bench, the pathway of microwaves is guided, so that low-power microwaves directly hit the sample and there is no dispersion of energy. Temperature of fixative did not increase after microwave irradiation. Fixation in the bench with either 4% PFA, or 1% PFA, for 20 minutes resulted in structural preservation of samples similar in quality as obtained with conventional fixation and in a similar or better level of antigen preservation. Therefore, controlling temperature and effective irradiation is crucial in order to obtain optimal structural and antigen preservation with microwave-enhanced fixation. The dramatic differences observed between microwave-irradiated samples and samples fixed in the same conditions without irradiation, strongly support the existence of specific effects of microwaves on fixation, independent from the mere heating of the samples.

Allium↗
PubMed1995

Design of sampling plans for mycotoxins in foods and feeds.

The control of the occurrence of mycotoxins in foods and feeds requires effective surveillance and quality control procedures which facilitate the identification and control of the mycotoxin problem respectively. Surveillance and quality control procedures involve a sequence of sampling, sample preparation, and analysis steps; and the integrity of the data produced by these procedures will be determined by the effectiveness of these steps. It is imperative that the sampling step is performed as accurately as possible so that the sample collected is representative of the batch of food or feed under investigation. Needless to say, the collection of a biased sample will completely invalidate the resultant analytical data. Most attempts to develop effective sampling protocols have focused upon the aflatoxins, since the majority of current regulations are concerned specifically with this group of mycotoxins. However, the design of effective sampling protocols has been severely hindered by the highly skewed distribution of the aflatoxins in foods and feeds. Studies already performed indicate that representative samples of commodities, composed of large particles (e.g., corn and oilseed kernels) should be 10 kg in weight, at least, and composed of approximately one hundred incremental samples. Similar studies have indicated that samples of oilseed cakes and meal, however, should be composed of fifty incremental samples which afford a composite sample of approximately 5 kg in weight.

Food Analysis↗
PubMed1993

Prenatal diagnosis by in situ hybridization on uncultured amniocytes: reduced sensitivity and potential risk of misdiagnosis in blood-stained samples.

Maternal cell contamination was assessed in 18 macroscopically blood-stained amniotic fluid samples from male fetuses. The samples were analysed by double-target fluorescent in situ hybridization (ISH) with Y and X chromosome-specific probes. The only sample with an aberrant karyotype (47,XY, + 18) was also analysed by hybridization with a chromosome 18-specific probe. An interpretation of extensive maternal cell contamination was made in two samples, one of which was the sample with trisomy 18. ISH with the chromosome 18-specific probe on this latter sample showed that the sensitivity of the ISH method for chromosome enumeration of uncultured amniotic fluid samples may be reduced in blood-stained samples. It was calculated that by using ISH for chromosome enumeration of the two extensively contaminated samples, a case of trisomy 21 might have been overlooked in both samples, while a case of trisomy 18 might only have been overlooked in one of the samples. It is concluded that ISH should not be used for chromosome enumeration of uncultured amniotic fluid samples that are macroscopically blood-stained without further technical developments.

Amniotic Fluid↗
PubMed2007

Evaluation of an on-target sample preparation system for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in conjunction with normal-flow peptide high-performance liquid chromatography for peptide mass fingerprint analyses.

Large-scale mass spectrometry (MS)-based proteomic analyses require high-throughput sample preparation techniques due to the increasing numbers of samples that make up a typical proteomics experiment. Moreover, extensive sample pre-treatment steps are necessary prior to MS acquisition for even the most rapid and robust MS-based proteomics methodology, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS followed by peptide mass fingerprinting (PMF) analysis. These include sample purification and fractionation, removal of digestion buffers or solvents, and spotting of sample with matrix onto the MALDI target. These multiple steps of time-consuming sample handling can result in high overall analysis costs and the likelihood of sample contamination and loss. In order to overcome some of these limitations in sample processing, we have investigated the use of a novel, simple, inexpensive 96-well elastomeric array that affixes to a MALDI target to create an on-target 96-well plate that accommodates a high solution volume (ca. 200 microL), thereby enabling the on-target processing of samples for MALDI-TOFMS. We explored several factors that influence MALDI sample preparation: type of matrix, solution volume, solution organic composition, solution drying rates and matrix/analyte co-crystallization methods. We also investigated the use of the 96-well elastomeric device for coupling MALDI-TOFMS analysis directly to high flow rate (1 mL/min) reversed-phase (rp)-HPLC. By developing an optimized, robust sample preparation protocol, we were able to obtain mass spectra with a high signal-to-noise ratio from peptide standards present at the 50-fmol level in large starting volumes of solution. PMF analyses were possible from 1-pmol and 500-fmol protein-digest standards. Coupling the device to high-flow HPLC (750 microL/min) yielded a robust and semi-automated means to obtain enhanced MALDI-TOFMS data at 500 ng of protein digest. These methodologies developed for this simple, on-target, elastomeric device show promise for streamlining the sample preparation process from HPLC to MALDI-MS.

Chromatography, High Pressure Liquid↗
PubMed1999

Evaluation of four sampling methods for determining exposure of children to lead-contaminated household dust.

Childhood exposure to lead has been demonstrated to result in health effects and lead-contaminated household dust is a primary exposure source. There is a need to establish reliable methods for sampling surfaces to determine levels of lead contamination. Three vacuums (HVS3, GS80, and MVM) and one wipe method were evaluated for the collection of household floor dust under field sampling conditions within a Superfund site and demographically similar control area. Side-by-side floor samples were taken from three locations within 41 randomly selected households between August and September 1995: a child's bedroom, primary play area, and primary entrance. Analysis was performed to assess the relative collection performance of each sampler, spatial distribution of lead within a household, and correlation of lead loading with observed blood lead level, and to determine if discrete or composites samples were more predictive of blood lead levels. Approximately 90% of the floor surfaces were carpeted. The rank order of sampling methods from greatest to lowest collection efficiency was HVS3 > GS80 > wipe > MVM. The HVS3 had the highest level of precision (CV=0.05), with the GS80 and wipe precisions 0.48 and 0.053, respectively. Lead loadings for samples collected in bedrooms and living areas and composite samples using the HVS3 and wipe methods were significantly correlated with blood lead levels. Correlations between blood lead levels and composite samples were stronger for the HVS3 (R(2)=0.33, P=0.003) and wipe (R(2)=0.25, P=0.002) methods than the respective discrete samples. Regression analysis indicated that a blood lead level of 10 microgram/dl corresponds to a carpet wipe sample geometric mean of 68 microgram/ft(2). For ongoing public health purposes, such as screening and clearance testing, use of the wipe sampling method is the most appropriate. This investigation supports findings by others that the present HUD risk levels for lead in floor wipe samples may not be adequate for reducing children's blood lead levels below 10 microgram/dl.

Child↗
PubMed2002

Optimal sampling strategy and core collection size of Andean tetraploid potato based on isozyme data - a simulation study.

Selection of an appropriate sampling strategy is an important prerequisite to establish core collections of appropriate size in order to adequately represent the genetic spectrum and maximally capture the genetic diversity in available crop collections. We developed a simulation approach to identify an optimal sampling strategy and core-collection size, using isozyme data from a CIP germplasm collection on an Andean tetraploid potato. Five sampling strategies, constant (C), proportional (P), logarithmic (L), square-root (S) and random (R), were tested on isozyme data from 9,396 Andean tetraploid potato accessions characterized for nine isozyme loci having a total of 38 alleles. The 9,396 accessions, though comprising 2,379 morphologically distinct accessions, were found to represent 1,910 genetically distinct groups of accessions for the nine isozyme loci using a sort-and-duplicate-search algorithm. From each group, one accession was randomly selected to form a genetically refined entire collection (GREC) of size 1,910. The GREC was used to test the five sampling strategies. To assess the behavior of the results in repeated sampling, k = 1,500 and 5,000 independent random samples (without replacement) of admissible sizes n = 50(50)1,000 for each strategy were drawn from GREC. Allele frequencies (AF) for the 38 alleles and locus heterozygosity (LH) for the nine loci were estimated for each sample. The goodness of fit of samples AF and LH with those from GREC was tested using the chi(2) test. A core collection of size n = 600, selected using either the P or the R sampling strategy, was found adequately to represent the GREC for both AF and LH. As similar results were obtained at k = 1,500 and 5,000, it seems adequate to draw 1,500 independent random samples of different sizes to test the behavior of different sampling strategies in order to identify an appropriate sampling approach, as well as to determine an optimal core collection size.

Journal Article↗
PubMed2006

Preservation of urine samples for metabolic evaluation of stone-forming patients.

Metabolic evaluation of stone-forming (SF) patients is based on the determination of calcium, oxalate, citrate, uric acid and other parameters in 24-h urine samples under a random diet. A reliable measurement of urinary oxalate requires the collection of urine in a receptacle containing acid preservative. However, urinary uric acid cannot be determined in the same sample under this condition. Therefore, we tested the hypothesis that the addition of preservatives (acid or alkali) after urine collection would not modify the results of those lithogenic parameters. Thirty-four healthy subjects (HS) were submitted to two non-consecutive collections of 24-h urine. The first sample was collected in a receptacle containing hydrochloric acid (HCl 6 N) and the second in a dry plastic container, with HCl being added as soon as the urine sample was received at the laboratory. Additionally, 34 HS and 34 SF patients collected a spot urine sample that was divided into four aliquots, one containing HCl, another containing sodium bicarbonate (NaHCO(3 )5 g/l), and two others in which these two preservative agents were added 24 h later. Urinary oxalate, calcium, magnesium, citrate, creatinine and uric acid were determined. Urinary parameters were also evaluated in the presence of calcium oxalate or uric acid crystals. Mean values of all urinary parameters obtained from previously acidified 24-h urine samples did not differ from those where acid was added after urine collection. The same was true for spot urine samples, with the exception of urinary citrate that presented a slight albeit significant change of 5.9% between samples in HS and 3.1% in SF. Uric acid was also not different between pre- and post-alkalinized spot urine samples. The presence of crystals did not alter these results. We concluded that post-delivery acidification or alkalinization of urine samples does not modify the measured levels of urinary oxalate, calcium, magnesium, creatinine and uric acid, and that the change on citrate was not relevant, hence allowing all parameters to be determined in a single urine sample, thus avoiding the inconvenience and cost of multiple 24-h urine sample collections.

Acids↗
PubMed1997

Distribution of Polychlorinated Naphthalene Congeners inEnvironmental and Source-Related Samples

Polychlorinated naphthalene (CN) congener profiles inenvironmental and source related samples were compared graphically and byprincipal component analysis. Samples investigated included biological,sediment, water, and air samples, technical polychlorinated biphenyl (PCB)and polychlorinated naphthalene (PCN) formulations, as well as municipalwaste incineration (MWI) fly ash and graphite electrode sludge. Biologicalsamples showed a preferential enrichment of planar, 1,3,5,7-substitutedtetra-, penta-, and hexachlorinated congeners and most of these samplesshowed profiles that displayed some similarity to those found in thetechnical PCB formulations. Sediment samples representing diffuse pollution,i.e., sediment samples from remote sites, showed an elevatedabundance of the planar hexa- and heptaCN congeners(1,2,3,4,6,7-/1,2,3,5,6,7- and 1,2,3,4,5,6,7-). The CN congener profile foundin these sediment samples and the two air samples were more similar to thetechnical PCB formulations than to the investigated MWI and graphite sludgesamples. Samples from three PCB contaminated lakes displayed similar congenerprofiles as Aroclor 1242, 1254 and Clophen A40. Two sediment samples and apike sample collected from the vicinity of a chloroalkali plant showedprofiles that were closely related to the investigated graphite electrodesludge sample. None of the environmental samples displayed profiles similarto low or medium chlorinated technical PCN (Halowax 1099, 1013, and 1014).

Journal Article↗
PubMed2001

Polymerase chain reaction in detection of Gymnodinium mikimotoi and Alexandrium minutum in field samples from southwest India.

Polymerase chain reaction (PCR) primers were constructed for the detection of two toxic dinoflagellate species, Gymnodinium mikimotoi and Alexandrium minutum. The primers amplified a product of expected size from cultured cells of G. mikimotoi and A. minutum. The species-specific primers targeting G. mikimotoi did not yield any product with a wide range of other cultured algae used as negative controls. Primers designed for A. minutum were species-group-specific since it PCR yielded a product from the closely related species A. ostenfeldii and A. andersonii, but not from other species of this genus tested. The confirmation of PCR products was performed by digestion of the products with restriction enzymes. Sensitivity analyses of the primers on DNA template from cultured cells was positive by PCR at a DNA template concentration of 1.5 x 10(-4) ng/microl (0.3 cells/L) for A. minutum, and at a DNA concentration of 2.5 x 10(-2) ng/microl (697 cells/L) for G. mikimotoi. The PCR method for detection of G. mikimotoi and A. minutum was applied on field samples collected with a plankton net. Gymnodinium mikimotoi could be detected in 11 field samples by microscopy, and all these field samples were positive by PCR. The cell counts of G. mikimotoi in simultaneously collected water samples ranged from 306 to 2077/L. Alexandrium minutum could be detected by microscopy in 3 different field samples. The cell counts in water samples collected at the same time as the net samples ranged from 115 to 1115 cells/L. Alexandrium minutum was detected by PCR in these field samples, with the exception of the sample displaying the lowest cell count (115 cells/L). Plankton samples that were negative by microscopy for any of the two target species were also negative by PCR. All the PCR products from field samples were confirmed by restriction enzyme digestion. The application of PCR-based detection of harmful algal bloom species for aquaculture and monitoring purposes in natural field samples is discussed.

Journal Article↗
PubMed2004

Sampling and interpolation of the a-wave of the electroretinogram.

UNLABELLED: This study was undertaken (a) to determine the minimum sampling frequency required to record a-waves evoked by flashes of very high energy without significant distortion and (b) to demonstrate that data sampled at a minimally adequate frequency can be interpolated to reconstruct the original waveform. METHODS: Dark-adapted ERGs from two anaesthetized macaque monkeys and an adult human were studied. Responses evoked by high-energy flashes that produced a-waves peaking as early as 5 or 6 ms after the flash were sampled at 5 kHz and transformed to obtain their discrete Fourier spectra. The amplitude of all spectral components above some cut-off frequency (e.g., 400 Hz) was then set to zero and the modified spectra transformed back into the time domain. The resulting computed responses, which contained no Fourier components above the cut-off frequency, were compared with the original recorded samples. To assess the validity of one method of interpolation, sample sets consisting of every fifth point of records sampled at 5 kHz (i.e., sets of 1 kHz samples) were subjected to Fourier transformation to give spectra with a frequency range of 0-500 Hz. These spectra were extended from 500 Hz up to a much higher frequency (e.g., 8 kHz) by adding zeros. The extended spectra were transformed back into the time domain to provide sets of interpolated samples at twice the chosen spectral frequency limit (i.e., 16 kHz). RESULTS: Removing all Fourier components above 400 or 500 Hz had no significant effect upon the leading edge or peak of the a-wave. However, removing Fourier components above 500 Hz gave rise to slight distortion of the oscillatory potentials (OPs) that appeared just after the a-wave peak on the leading edge of the b-wave. Except for this small distortion, the original 5 kHz data samples corresponded very well with the interpolated curves that had been generated as described above from a 1 kHz subset of the samples. This provides further confirmation that dark-adapted ERG a-waves evoked by flashes of up to about 50,000 sc. Td sc do not contain Fourier components with frequencies above 500 Hz. CONCLUSION: Human and macaque a-waves are completely represented by 1 kHz samples and Fourier methods can be used to reconstruct the original continuous waveform. However, to capture the OPs with complete fidelity, a higher sampling rate is necessary.

Adult↗