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1.
The development of mass spectrometry-based leak detection for pharmaceutical container integrity was undertaken to provide an alternative to microbial challenge testing. Standard 10-mL vials were modified to contain pinholes (0.5 to 10 microns) by affixing micropipettes with epoxy into 2-mm vial side wall holes. The absolute leak rate was determined using vials that were sealed in a tracer (helium) environment with butyl rubber stoppers and crimps. Alternatively leak rates were determined using vials that were sealed in room air and exposed to tracer under pressure (charging or bombing). Tracer leak rates were measured with mass spectrometry leak rate detectors. The absolute leak rate was correlated the squared nominal leak radius which suggested that the mode of gas flow through the glass pipette leaks was more turbulent than viscous even at low leak rates typically associated with viscous flow. The minimum observed absolute leak rate was about 10(-6.6) std cc/sec and was likely due to helium permeation through the rubber stoppers. Heat-stressed rubber stoppers did not affect the baseline absolute leak rate. Adsorption of helium tracer to the test unit surfaces was found to confound baseline leak rate measurement reliability but was eliminated as a source of variation by exposing the test units to ambient air for > or = 12 hours. The absolute leak rate and the leak rate measured after charging were related in a mathematically predictable way.  相似文献   

2.
The shell vial assay is a sensitive, rapid test for the detection of cytomegalovirus (CMV) in a variety of specimens. The sensitivity of this assay is dependent on a number of factors including the antibodies used for immunostaining. Monoclonal antibodies to the CMV major immediate-early antigen (p72) from Chemicon (MAB810) and Dupont (NEA-9221) were assessed side by side in duplicate vials on 4,388 specimens from a patient population consisting of > 90% organ transplant recipients. A total of 240 specimens (5.5%) were CMV positive in either one or both vials. Positivity rates were variable across different specimen types but highest (12.9%) in urine specimens. Of the positive specimens, 175 (72.9%) tested positive in both vials, 43 (17.9%) tested positive in the Chemicon-stained vial only, and 22 (9.2%) tested positive in the Dupont-stained vial only (P < 0.01, McNemar's chi-square test). This gave an overall positivity rate of 5.0% for Chemicon antibodies and 4.5% for Dupont. There was no difference in the fluorescent focus counts produced by the two antibody sets. It is concluded that use of the Chemicon antibodies provides increased sensitivity of detection of CMV in the shell vial assay above that afforded by the Dupont antibody.  相似文献   

3.
Five reference laboratories were established in Pakistan for monitoring cholinesterase (ChE) activities of workers exposed to organophosphorus compounds. ChE activities were determined by the Michel and tintometric method. Observations of ChE activities were made during two malaria seasons. The first season showed that although a significant depression of cholinesterase occurred among some of the workers, the ChE activities of workers were within the normal range during the following season. The reason for the difference is discussed. Similar studies were undertaken in Haiti. Mean and standard deviations (SD) were calculated for comparison of the tintometric versus the Michel method. The data show a correlation between the methods. For further evaluation of the tintometric method, organophosphorus and oxon analog inhibition of cholinesterase were determined in vitro. The tabulated data show that the tintometric method is adequate for determining whether a worker is exposed to dangerous amounts of insecticides.  相似文献   

4.
The application of a newly developed thermal desorption method for the analysis of workplace air to the analysis of polar compounds is reported. The method was validated for both pumped and diffusive sampling of test gases containing polar volatile organic compounds (esters, alcohols, ketones or aldehydes) on adsorption tubes and subsequent analysis of these tubes. Carbosieve SIII, Carboxen 569, Carbopack B and Tenax TA were used as solid adsorbents. Analysis was performed by thermal desorption of the analytes from the adsorbent tubes followed by gas chromatography-flame ionisation detection (GC-FID). It could be demonstrated that thermal desorption-GC-FID is feasible also for the analysis of polar compounds and that problems arising from the high concentration levels of some analytes in workplace air could be solved.  相似文献   

5.
A simple, inexpensive modification in the elevator mechanism of a liquid scintillation counter allows for radioimmunoassay tubes (12 X 75 mm) containing the antibody-bound radioligand to be counted directly, thereby obviating the need for counting vials and considerably decreasing the amount of fluor used. Furthermore, because assay and counting are all done in the same tube, the modification shortens assay time and increases precision by avoiding what is often the most time consuming and inaccurate step, the transfer of antibody-bound radioligand to a counting vial. We find that use of this modification results in about a 75% decrease in the overall cost for materials and a substantial decrease in assay time.  相似文献   

6.
Organophosphates, a widely used class of insecticidal compounds, have been shown to cross the placental barrier, and thus potentially affect the developing fetus. This study compared the maternal and fetal effects, including cholinesterase inhibition, following gestational exposure to six organophosphates: tribufos, oxydemeton-methyl, azinphos-methyl, fenamiphos, isofenphos, and fenthion in the Sprague-Dawley rat. All test compounds were administered via oral gavage on gestation days 6-15. Maternal cholinesterase activities (plasma, PChe; erythrocyte, RChe; and brain, BChe) were measured on gestation days 16 and 20, and fetal brain cholinesterase activity was measured on gestation day 20. Effects on gestational parameters (clinical signs, food consumption, and body weight) in adult rats, when observed, were only observed at the highest dose tested for each compound. The inhibition of maternal cholinesterase activities associated with these clinical findings was, for all compounds, always greater than 20%. Moreover, cholinesterase activities were inhibited at dose levels below that which elicited clinical effects. Statistically significant inhibition of at least two of the three cholinesterase enzymes (PChe, RChe, or BChe) was observed on gestation day 16, 24 h following exposure, with all of the organophosphates tested. By gestation day 20, the inhibition of cholinesterase activity was reduced; however, the high dose for all test compounds (except BChe in fenamiphos-treated dams) continued to demonstrate statistically significant inhibition of RChe and BChe. Despite significantly affected cholinesterase activity in the dams, no remarkable effects on fetal BChe were observed with any test compound. No embryotoxicity or teratogenicity were observed with any of the test compounds. These results demonstrate that for the six organophosphates tested: (1) inhibition of maternal cholinesterase activity was the most sensitive indicator of organophosphate exposure; (2) the level of cholinesterase inhibition associated with clinical findings was always greater than 20%; and (3) no effect on fetal cholinesterase activity (BChe) was observed, even at dose levels that continued to demonstrate significant inhibition of maternal cholinesterase activity.  相似文献   

7.
Detection of enteroviruses and adenoviruses mainly in fecal specimens by rapid culture with inoculation onto cell monolayers in flat-bottom tubes by centrifugation and immunofluorescence staining with genus-specific monoclonal antibodies was compared with that by the conventional virus isolation procedure. For both conventional culture and shell vial culture human lung fibroblast cells and tertiary monkey kidney cells were used. For enterovirus detection, 979 clinical specimens (916 stool specimens, 56 cerebrospinal fluid specimens, and 7 nasopharyngeal swabs) were used. Conventional culture detected 74 enterovirus isolates. A cytopathic effect compatible with the presence of an enterovirus after 3 days of incubation occurred in 25 of the 74 (34%) specimens that eventually became positive. The detection rate for enteroviruses by rapid cell culture after 2 to 3 days of incubation was 42 of 74 (57%). The genus-specific enterovirus monoclonal antibody did not react with strains of echovirus types 22 and 23 or enterovirus type 71. Rapid cell culture for the detection of adenoviruses was performed with 567 clinical specimens (536 stool specimens, 25 cerebrospinal fluid specimens, and 6 miscellaneous specimens), in which 42 adenoviruses were found by conventional culture. Nine of the 42 (21%) adenovirus isolates were detected by conventional culture within 3 days after inoculation, whereas 21 (50%) were found by rapid cell culture within 2 to 3 days. Only two of the nine specimens found to be positive for the enteric adenovirus type 41 by conventional culture as well by a type-specific enzyme-linked immunosorbent assay (ELISA) tested positive by rapid cell culture. In conclusion, the rapid shell vial assay allows the early detection and identification of enteroviruses and adenoviruses in clinical specimens but is markedly less sensitive than the conventional isolation procedure according to the eventual results of the conventional isolation procedure. Conventional cell culture remains a prerequisite for serotyping of enteroviral isolates. On the basis of the results for adenovirus type 41, the rapid detection of adenoviruses was not considered to be useful for the detection of clinically relevant adenoviruses in fecal samples.  相似文献   

8.
A fiber-optic microbial biosensor suitable for direct measurement of organophosphate nerve agents was developed. The unique features of this novel microbial biosensor were the recombinant Escherichia coli cells expressing the enzyme organophosphorus hydrolase on the cell surface and the optical detection of the products of enzyme-catalyzed organophosphate hydrolysis. The use of cells with the metabolic enzyme expressed on the cell surface as a biological sensing element provides advantages of no resistance to mass transport of the analyte and product across the cell membrane and low cost due to elimination of enzyme purification, over the conventional microbial biosensors based on cells expressing enzyme intracellularly and enzyme-based sensors, respectively. The use of an optical transducer allows the detection of different organophosphates in a mixture, presently not feasible with acetylcholinesterase-based biosensors. E. coli cells expressing organophosphorus hydrolase (OPH) on the cell surface were immobilized in low melting temperature agarose on a nylon membrane and attached to the common end of a bifurcated fiber-optic bundle. OPH-expressing E. coli cells catalyzed the hydrolysis of organophosphorus pesticides to form stoichiometric amounts of chromophoric products that absorb light at specific wavelengths. The backscattered radiation of the specific wavelength incident light was measured using a photomultiplier detector and correlated to the organophosphate concentration. The best sensitivity and response time were obtained using a sensor constructed with 1.5 mg of cells operating in pH 9, 50 mM HEPES buffer with 100 mM NaCl and 0.05 mM CoCl2 at 30 degrees C. At optimized conditions, the biosensor measured paraoxon, parathion, and coumaphos pesticides with high selectivity against triazine and carbamate pesticides in approximately 10 min. The lower detection limits were 3 microM for paraoxon and parathion and 5 microM for coumaphos. When stored in the buffer at 22 degrees C, the biosensor was stable for over a 1-month period and showed no decline in the response for over 75 repeated usages. The new fiber-optic microbial biosensor is an ideal tool for on-line monitoring of the detoxification process for organophosphate pesticides-contaminated wastewaters but may not be suitable for environmental monitoring.  相似文献   

9.
The aim of the study was to evaluate the morphological and functional status of the liver in acute, oral cholinesterase inhibitors poisoning using static scintigraphy, hepatography and measurements of chosen enzymes activity. Considering the different clinical picture of cholinesterase inhibitors poisonings in people, it was necessary to estimate the poisoning severity and its dependence on the frequency and intensification of the liver lesion. Under examination there were 37 cholinesterase inhibitors orally poisoned patients, treated at the Department of Toxicology in the years 1992-1995. The examined group comprised 7 women (19%) and 30 men (81%). Organophosphate compounds poisoning was noted in 14 patients, and carbamates poisonings in 23 patients. The reference group comprised 30 healthy men aged 24 to 59 years not exposed to hepatotoxic agents. More than 90% of patients were classified as severe poisoned. Any fatal case was not noted. A differently intensified pathological changes of the liver dependent on age and poisoning severity were found in 97.2% of patients and their frequency was significantly higher than in the control group. Hepatographic picture revealed in 96.6% of cases the liver lesion. Hepatographic picture of the liver was also dependent on poisoning severity. The higher activity of AST, ALT, AP and higher bilirubin concentration in blood were noted in poisoned men compared to the control group. Control scintigraphic examination revealed a considerable improvement in the intensification of the liver scintigraphic picture in 40% of the patients and a higher intensification in 13% of the subjects. In 46.6% of the patients the intensification of scintigraphic changes remained at the same level. Considering arbitrary criteria for the degree of the liver lesion, the improvement in the intensification of hepatographic changes was noted in 42.8% of the patients; the intensification of the liver lesion was not noted even in one case. Analyzing the percentage of the liver lesion for each individual patient, improvement was noted in 92.8% of the examined patients, and the changes with the same level of intensification in 7.2%. Deterioration was not noted at all. Conclusion: The liver scintigraphy and hepatography combined with biochemical analysis allows to assess the liver condition in acute cholinesterase inhibitors poisoning.  相似文献   

10.
A case of the foreign body due to coring from the rubbercap of a 50 ml Diprivan vial was reported. Furthermore, how the fragment is made and how to decrease the incidence are discussed. The incidence of coring should decrease by improvement of the needle and by sticking vials vertically. However, even if the improvement of the rubber, needles and sticking technique is achieved, the occurrence of coring can not be eliminated completely. The best way to avoid coring will be to furnish "50 ml syringes containing Diprivan" commercially.  相似文献   

11.
Standard genotoxicity tests are often inappropriate for testing new biological entities, in particular for recombinant proteins which are nature-identical. Arguments that these may contain mutagenic impurities are not substantiated; however, we have produced evidence that such impurities would be detected amidst a vast excess of protein. Concerns that human patients receiving therapy may be at risk from higher-than-physiological levels of proteins are also somewhat theoretical. However, it is apparent that genotoxicity testing will be required for these products for the time being, even if pragmatic approaches reduce the battery of in vitro tests to Ames and chromosomal aberrations only, and reduce the top dose in vivo to 1000x the human therapeutic dose. There is a number of physical and chemical properties of proteins that demand special approaches to methodology if the tests are to produce accurate results. The potential for adsorption to certain forms of glass and plastic means special care must be taken in dissolving and diluting test solutions; adherence to filters means special low protein binding, non-pyrogenic filters should be used for sterilisation of test solutions, where this is necessary; freeze-dried powders aliquotted in multiple vials should be dissolved in minimal solvent and cascaded from vial to vial rather than trying to empty the solid contents for bulk weighing. As proteins are often supplied in solution, in order to achieve sufficiently high test concentrations, it may be necessary to resuspend test bacteria/cells in the test solutions for short periods of time before centrifuging and resuspending in selective or growth media.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

12.
The widespread use of organophosphate pesticides creates the possibility of excessive exposure of migrant farm workers to these compounds. Blood cholinesterase determinations were used to compare the organophosphate pesticide exposure of 57 migrant farm workers with that of 35 controls. Frequently reported symptoms of the farm workers which might be related to pesticide exposure were also studied, including headaches, dizziness, loss of weight, nausea, and a general feeling of weakness or loss of energy. Significantly depressed cholinesterase activities were found in the farm workers, with 10.5% of the farm workers having values below the lower limit of normal. There was no significant relationship between frequently reported symptoms of the farm workers and depressed cholinesterase levels.  相似文献   

13.
The objective of this study was to evaluate manometric temperature measurement as a non-invasive method of monitoring product temperature during the primary drying phase of lyophilization. This method is based on analysis of the transient response of the chamber pressure when the flow of water vapor from the chamber to the condenser is momentarily interrupted. Manometric temperature measurements (MTM) were compared to product temperature data measured by thermocouples during the lyophilization of water, mannitol, lactose and potassium chloride solutions. The transient pressure response was mathematically modeled by assuming that four mechanisms contribute to the pressure rise: 1) direct sublimation of ice through the dried product layer at a constant temperature, 2) an increase in the temperature at the sublimation interface due to equilibration of the temperature gradient across the frozen layer, 3) an increase in the ice temperature due to continued heating of the frozen matrix during the measurement, and 4) leaks in the chamber. Experimental transient pressure response data were fitted to an equation consisting of the sum of these terms containing three variables corresponding to the vapor pressure of ice, product resistance to vapor flow, and the vial heat transfer coefficient. Excellent fit between the mathematical model and the experimental data was observed, and the value of the variables was calculated from the measured transient pressure response by a least squares method. The product temperature measured by MTM, which measures the temperature at the sublimation interface, was compared with product temperature measured by thermocouples placed in the bottom center of the vials. Manometrically measured temperatures were consistently lower than the thermocouple measurements by about 2 degrees C, this difference being largely accounted for by the temperature gradient across the frozen layer. The resistance of the dried product to mass transfer calculated from MTM was found to agree reasonably well with values measured by a direct vial technique. Product resistance was observed to increase with increasing solute concentration, and to increase continuously as the depth of the dried product layer increases for mannitol and potassium chloride. For lactose, product resistance increases continuously with thickness up to the onset of collapse, at which point the product resistance becomes essentially independent of depth. Scanning electron microscopy was used to explain this observation based on changes in morphology of the solid. The vial heat transfer coefficients obtained from regression analysis were on the order of 10(-3)-10(-4) cal.sec-1. degrees C-1; however, the scatter in the vial heat transfer coefficient data prevents the method from being used for accurate measurement of the vial heat transfer coefficient. The results of the study show that the manometric method shows promise as a process development tool and as an alternative method of in-process product temperature measurement during primary drying.  相似文献   

14.
Several routinely employed diagnostic methods were analysed for their usefulness in aiding an early and rapid diagnosis of human cytomegalo-virus infection in immunocompromised patients. Clinical samples obtained during an 18-month period were examined by conventional culture, the shell vial method, detection of pp65 antigen and the polymerase chain reaction. Detection of pp65 antigen in peripheral leukocytes was the most useful method for rapid detection of infection at an early stage. Results of other rapid detection methods, the shell vial method and the polymerase chain reaction, gave useful support, while results obtained by conventional culture were not available until after the initiation of therapy. Only a small proportion of serological tests provided useful information for determining whether to treat the patient.  相似文献   

15.
OBJECTIVE: To identify risk factors for polymicrobial bloodstream infections (BSIs) in neonatal intensive care unit (NICU) patients during an outbreak of BSIs. DESIGN: During an outbreak of BSIs, we conducted a retrospective cohort study, assessed NICU infection control practices and patient exposure to NICU healthcare workers (HCWs), and obtained cultures of the environment and HCW hands. PATIENTS: During the period May 3 to 7, 1996, 5 infants contracted BSIs caused by both Enterobacter cloacae and Pseudomonas aeruginosa, and one infant contracted a BSI caused by E cloacae only. For each pathogen, all isolates were identical on DNA typing. RESULTS: Infants exposed to the following were more likely than nonexposed infants to have BSI: umbilical venous catheters (6/14 vs 0/7, P = .05), total parenteral nutrition given simultaneously with a dextrose/electrolyte solution (6/12 vs 0/9, P = .02), or one HCW (5/7 vs 1/13, P = .007). Neither environmental nor HCW hand cultures yielded the outbreak pathogens. Quality control cultures of intravenous solution bags were negative. CONCLUSIONS: We speculate that a dextrose multidose vial became contaminated during manipulation or needle puncture and that successive use of this contaminated vial for multiple patients may have been responsible for BSIs. Aseptic techniques must be employed when multidose vial medications are used. Single-dose vials should be used for parenteral additives whenever possible to reduce the risk of extrinsic contamination and subsequent transmission of nosocomial pathogens.  相似文献   

16.
A radio gas-chromatographic system consisting of a gas chromatograph, combustion furnace, proportional counter and guard scintillation counter has been studied for the analysis of trace amounts of 14C-labelled organophosphorus insecticides. The proportional detector is a concentration sensitive detector requiring strict flow control. Sensitivity depends on the concentration of the quenching gas used. Using 14C-ethyl parathion and 14C-metamidophos as model compounds a linear range of 3500-5000 was measured. A sensitivity of 0.98 cpm/pCi and a detection limit of 24pCi were obtained which allows the detection of 0.33 ng if the specific activity of the insecticide is 21.5 mCi/mMol. The system must be decontaminated in order to achieve optimum detection levels by passage of oxygen through the combustion tube and by periodically injecting the corresponding non-labelled compound into the column. A solvent effect was found to interfere in the analysis due to the sudden production and expansion of CO2. This effect was eliminated by choosing column conditions to prevent elution near the solvent. The system was tested by analyzing a mixture of 37 ppb and 23 ppb of ethyl parathion and its oxygen analog in orange juice.  相似文献   

17.
Liquid chromatography-mass spectrometry (LC-MS) with a particle beam (PB) interface is used to separate and identify a group of pesticides. The mass spectra obtained under the different ionization modes, electron ionization (EI) and positive and negative chemical ionization (PCI and NCI) are compared. The operating conditions under each mode, determined by studying the influence on the ion abundance of the ion source temperature of the EI mode, and the gas pressure and ion source temperature in the methane CI were optimized. EI was more sensitive than PCI and NCI and of the latter two modes, NCI gave higher responses, especially for organophosphorus compounds. When on-line solid-phase extraction-LC-PB-MS was applied to real samples, limits of detection in full scan mode were in the range of 0.5 and 10 micrograms l-1 for EI. The analysis of real samples by on-line solid-phase extraction-LC-PB-MS enabled EI detection of one of the pesticides studied and confirmation by PCI and NCI. The combined EI/CI information also enabled the detection of some non-target compounds.  相似文献   

18.
Glucose utilization of four cerebral cortex and 35 subcortical regions (CGU) was analyzed in three models of cholinergic seizures induced by the following compounds: 1) soman (pinacolylmethylphosphonofluoridate) an organophosphorus cholinesterase inhibitor, 100 microg/kg SC after pretreatment with pyridostigmine 26 microg/kg IM (n = 6); 2) physostigmine, a carbamate cholinesterase inhibitor, 1.31 mg/kg infused IV over 75 min (n = 6); and 3) pilocarpine, a direct cholinergic agonist, 30 mg/kg SC (n = 6). Physostigmine and pilocarpine were preceded by 3 mmol/kg LiCl IP 20 hrs earlier. Animals injected with saline SC (n = 6) were used as controls. Step-wise discriminant analysis successfully classified 100% of the cases into the four experimental groups with data from only six regions. Pyridostigmine-soman induced the most widespread and greatest increases in CGU. More restricted and lower levels of activation were observed with Li-pilocarpine while Li-physostigmine induced significant increases in CGU only in globus pallidus, entopeduncular nucleus, and substantia nigra. These three regions, which are functionally related, were also activated in the other two models of cholinergic convulsions and may represent the initial step in cholinergic activation of the CNS. Li-pilocarpine failed to activate most of the brainstem and the superior colliculus. All cortical regions were activated by Li-pilocarpine and pyridostigmine-soman, while they were inhibited by Li-physostigmine. This phenomenon may be due in part to the lack of activation with physostigmine of the basal forebrain nuclei (lateral septum, medial septum, vertical and horizontal limbs of the diagonal band, and substantia innominata) resulting in a decreased drive of cortical metabolism.  相似文献   

19.
A total of 663,533 colonies from 72 dairy and meat sources showed a detection rate of 0.2% for bacteriocin producers using direct plating techniques. A further 83,000 colonies from 40 fish and vegetable sources showed a detection rate of 3.4% for bacteriocin producers using selective enrichment procedures. A collection of seven purified isolates showing a different host spectrum of bacteriocin activity and with the ability to produce bacteriocins in broth culture were compared with nisin and pediocin (with respect to their inhibitory activity, determined by the critical dilution method), against various indicator bacteria in agar and broth. The sensitivity of Listeria species to various bacteriocins was influenced by the agar and broth test systems used. A Lactobacillus curvatus strain was found to be the most suitable indicator for quantitating antimicrobial effects of all the bacteriocins investigated in both agar and broth test systems. The bacteriocin-producing isolates were characterized by biochemical reactions and DNA restriction enzyme profiles and taxonomic identification revealed species of Lactobacillus, Carnobacterium and Lactococcus assigned on the basis of 16S rDNA sequences.  相似文献   

20.
Incorporating resins in blood culture media can effectively reduce the activities of several antibiotics. It was shown that the activities of some generally used antibiotics decreased by 80 to 90% within 2 h in Bactec Plus Aerobic/F resin-containing culture medium. Bactec vials containing resins were still found to be positive for bacteria when antibiotics were present. The addition of beta-lactamase shortened the detection time irrespective of the presence of resins.  相似文献   

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