Technical note: a device for obtaining time-integrated samples of ruminal fluid.
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A device was adapted to allow for time-integrated sampling of fluid from the rumen via a cannula. The sampler consisted of a cup-shaped ceramic filter positioned in the ventral rumen of a cannulated cow and attached to a tube through which fluid entering the filter was removed continuously using a peristaltic pump. Rate of ruminal fluid removal using the device was monitored over two 36-h periods (at 6-h intervals) and was not affected (P > .05) by time, indicating that the system was not susceptible to clogging during this period. Two cows having ad libitum access to a totally mixed ration were used in a split-block design to evaluate the utility of the system for obtaining time-integrated samples of ruminal fluid. Ruminal fluid VFA concentration and pattern in samples collected in two replicated 8-h periods by the time-integrated sampler (at 1-h intervals) were compared with composite samples collected using a conventional suction-strainer device (at 30-min intervals). Each 8-h collection period started 2 h before or 6 h after feeding. Results indicated that total VFA concentration was not affected (P > .05) by the sampling method. Volatile fatty acid patterns were likewise unaffected (P > .05) except that acetate was 2.5% higher (P < .05) in samples collected 2 h before feeding and valerate was 5% higher (P < .05) in samples collected 6 h after feeding by the suction-strainer device. Although significant, these differences were not considered physiologically important. We concluded that use of the ceramic filter improved the sampling of ruminal fluid by simplifying the technique and allowing time-integrated samples to be obtained. (+info)
Reactivity of monoclonal antibodies EG1 and EG2 with eosinophils and their granule proteins.
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Use of the murine monoclonal antibodies EG1 and EG2 has been based on the assumption that EG2 recognizes activated eosinophils. We examined the reactivity of EG1 and EG2 with eosinophil cationic protein (ECP), eosinophil-derived neurotoxin (EDN), and stimulated and nonstimulated eosinophils from normal donors. By radioimmunoassay, EG1 recognized only ECP, whereas EG2 recognized both ECP and EDN. By Western blot, EG1 reacted with ECP, EG2 reacted with both ECP and EDN, but EG2 could not distinguish between lysates of stimulated and nonstimulated eosinophils. By immunofluorescence, EG1 and EG2 at 20 microg/mL stained 95-100% of nonstimulated eosinophils, regardless of fixative; EG1 and EG2 at 0.1 microg/mL stained 61-90% of acetone- and paraformaldehyde-fixed and only 5-21% of methanol-fixed nonstimulated eosinophils. Thus, the reactivity of EG1 and EG2 with eosinophils depends on the method of fixation and antibody concentration; and EG2, in contrast to previous reports, cannot reliably discriminate between resting and activated eosinophils. (+info)
Arsenic speciation in humans and food products: a review.
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Although acute intoxication has become rare, arsenic (As) is still a dangerous pollution agent for industrial workers and people living in the vicinity of emission sources. In humans, only inorganic As is toxic; organic forms present in large amounts in the environment are nontoxic. It is therefore important to be able to differentiate one group from the other using appropriate speciation methods. The authors review the present knowledge of the distribution of As in humans and food products. The three steps of the speciation methods (sample preparation, species separation, and detection) are described. For liquid samples, a clean-up step (C18 cartridge extraction, dilution, or freezing) is necessary to eliminate proteins and salts from the matrix. For solid organic samples, the first step consists of the digestion of tissues followed by solvent extraction sometimes coupled with a C18 extraction. The separation of As species is accomplished by different high-performance liquid chromatography (HPLC) methods (ion-exchange, ion-pairing, and micellar liquid chromatography). The detection methods are compatible with HPLC and are able to detect As species in the microgram-per-liter range. Inductively coupled plasma (ICP) atomic emission spectrometry is more frequently used, but suffers from interference by organic solvents in the mobile phases. Atomic absorption spectrometry methods give sensitivities of the same order. ICP-mass spectrometry has the advantage of specificity and can be 100- to 1000-fold more sensitive than previous methods. (+info)
Field testing of a personal size-selective bioaerosol sampler.
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Existing samplers for the collection of bioaerosols have been designed with the aim of maintaining biological stability of the collected material, and in general do not select particles in accordance with international conventions for aerosol sampling. Many have uncharacterised sampling efficiencies and few are designed as personal samplers. If standard personal dust samplers are used for bioaerosols the viability of collected microorganisms may be compromised by dehydration. The objective of this study was to evaluate a novel personal bioaerosol sampler designed to collect the inhalable dust fraction and further subdivide the sample into thoracic and respirable fractions. The new sampler was tested to see whether it enhanced the survival of the collected microorganisms, and was assessed for ease of use in the field and in subsequent laboratory analyses. A number of occupation-related field sites were selected where large concentrations of bioaerosols were to be expected. The prototype sampler was found to be simple to use. Analysis could be carried out with similar efficiency either with all three fractions together for a total count, or separately for size selective data. The sampler performed at least as well as the standard IOM filter method but with the added advantage of size fractionation. The field trials showed that for sampling periods lasting several hours, microorganism survival within the sampler was adequate for culture and identification of the organisms present. This new sampler is now commercially available. In addition to bioaerosol sampling, the principle of size selective sampling using porous foams can be applied to other occupational hygiene problems, and also to indoor air monitoring of PM10 and PM2.5 concentrations. (+info)
Sample preparation and storage can change arsenic speciation in human urine.
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BACKGROUND: Stability of chemical speciation during sample handling and storage is a prerequisite to obtaining reliable results of trace element speciation analysis. There is no comprehensive information on the stability of common arsenic species, such as inorganic arsenite [As(III)], arsenate [As(V)], monomethylarsonic acid, dimethylarsinic acid, and arsenobetaine, in human urine. METHODS: We compared the effects of the following storage conditions on the stability of these arsenic species: temperature (25, 4, and -20 degrees C), storage time (1, 2, 4, and 8 months), and the use of additives (HCl, sodium azide, benzoic acid, benzyltrimethylammonium chloride, and cetylpyridinium chloride). HPLC with both inductively coupled plasma mass spectrometry and hydride generation atomic fluorescence detection techniques were used for the speciation of arsenic. RESULTS: We found that all five of the arsenic species were stable for up to 2 months when urine samples were stored at 4 and -20 degrees C without any additives. For longer period of storage (4 and 8 months), the stability of arsenic species was dependent on urine matrices. Whereas the arsenic speciation in some urine samples was stable for the entire 8 months at both 4 and -20 degrees C, other urine samples stored under identical conditions showed substantial changes in the concentration of As(III), As(V), monomethylarsonic acid, and dimethylarsinic acid. The use of additives did not improve the stability of arsenic speciation in urine. The addition of 0.1 mol/L HCl (final concentration) to urine samples produced relative changes in inorganic As(III) and As(V) concentrations. CONCLUSIONS: Low temperature (4 and -20 degrees C) conditions are suitable for the storage of urine samples for up to 2 months. Untreated samples maintain their concentration of arsenic species, and additives have no particular benefit. Strong acidification is not appropriate for speciation analysis. (+info)
A prospective, randomized, double-blind and placebo-controlled study to assess the efficacy of paracervical block in the pain relief during egg collection in IVF.
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The role of paracervical block in the pain relief during egg collection in in-vitro fertilization (IVF) is still not confirmed. In this prospective, double-blind and placebo-controlled study, 135 patients undergoing egg collection in their first IVF cycle were randomized to receive 10 ml of 1.5% lignocaine (group A) or normal saline (group B) in the paracervical block and no local injection (group C). No differences were seen among the groups in the demographic data, the ovarian response, the duration of egg collection, the number of follicles punctured, the pregnancy rates and the pain levels related to blood taking, scanning and insertion of an i.v. cannula. All patients experienced similar pain scores for vaginal puncture but patients in group A experienced significantly less abdominal pain during egg collection, compared with those in group B and group C (P = 0.009 and P = 0.001 respectively; Mann-Whitney U-test). When lignocaine was used, the abdominal pain scores were reduced by 38.9 and 51.4% compared with placebo and no local injection respectively. We recommend that paracervical block with lignocaine should be used in conjunction with i.v. sedation/analgesia during egg collection performed through the transvaginal route under ultrasound guidance (TUGOR) to reduce the pain of the procedure. (+info)
Reproductive capacity of spermatozoa from men with testicular failure.
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Controversial reports have been published about the influence of sperm source and of the underlying testicular pathology on success rates of intracytoplasmic sperm injection (ICSI). In this controlled study, ICSI treatment cycles with testicular spermatozoa from men with obstructive and non-obstructive azoospermia were compared with ICSI ejaculated sperm cycles with semen parameters < or = 5 x 10(6)/ml and < or = 10% progressive motility. The control cases were matched for female age, rank of trial, female basal follicle-stimulating hormone serum concentrations and close proximity to the study group's procedure. The fertilization, cleavage, pregnancy and abortion rates were similar in matched groups irrespective of the type of azoospermia. However, the implantation rate in the non-obstructive azoospermic patient group was significantly lower than that in the matched ejaculated sperm group (13.4% versus 26%, P = 0.05). On the other hand, no impairment of the implantation rate was observed in the obstructive azoospermic patient group. These data show that testicular pathology has a negative impact on reproductive performance of testicular spermatozoa, resulting in a decreased implantation potential without any apparent effect on fertilization and early preimplantation development. (+info)
Postal urine specimens: are they a feasible method for genital chlamydial infection screening?
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BACKGROUND: A United Kingdom (UK) screening programme for Chlamydia trachomatis has recently been announced. Pilot projects involving the opportunistic testing of women attending health facilities are due to commence in several sites. There is a danger that this approach will fail to obtain adequate population coverage. The alternative--true systematic population screening--is generally assumed to be unfeasible. Studies in Denmark using postal urine specimens have challenged this assumption. No such studies have been reported from the UK. AIM: To assess the potential of urine specimens sent by post as the basis for a UK population screening strategy for genital chlamydial infection. METHOD: Two hundred patients (100 men, 100 women) aged 18 to 45 years were randomly sampled from the list of one urban group practice. Subjects were mailed an explanatory letter, a urine sample container, a sexual lifestyle questionnaire, and a prepaid return envelope. Non-responders were contacted by telephone; persistent non-responders were visited at home. Samples were tested for Chlamydia by DNA amplification and enzyme immunoassay. RESULTS: Sixty-four (32%) subjects were no longer living at their GP registered address. Of the remaining 136, 126 (93%) responded to the survey and 113 (83%) accepted the request for a urine sample and completed a questionnaire. Acceptance rates were similar for men and women and across age groups. Four samples (3%) were Chlamydia positive. CONCLUSION: Home mailed urine specimen collection in conjunction with a self-completed postal questionnaire is feasible. This could provide a viable basis both for determining population Chlamydia prevalence and for a UK Chlamydia population screening strategy. Overall cost effectiveness of such a strategy will depend on the cost of the test used. Comparative performance characteristics of the different currently available tests in this setting have yet to be fully determined. (+info)