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Confocal microscopy and microbial viability detection for food research.

Confocal microscopy offers several advantages over other conventional microscopic techniques as a tool for studying the interaction of bacteria with food and the role of food microstructure in product quality and safety. When using confocal microscopy, samples can be observed without extensive preparation processes, which allows for the evaluation of food without introducing artifacts. In addition, observations can be made in three dimensions without physically sectioning the specimen. The confocal microscope can be used to follow changes over a period of time, such as the development of the food structure or changes in microbial population during a process. Microbial attachment to and detachment from food and food contact surfaces with complex three-dimensional (3-D) structures can be observed in situ. The fate of microbial populations in food system depends on processing, distribution, and storage conditions as well as decontamination procedures that are applied to inactivate and remove them. The ability to determine the physiological status of microorganisms without disrupting their physical relationship with a food system can be useful for determining the means by which microorganisms survive decontamination treatments. Conventional culturing techniques can detect viable cells; however, these techniques lack the ability to locate viable cells in respect to the microscopic structures of food. Various microscopic methods take advantage of physiological changes in bacterial cells that are associated with the viability to assess the physiologic status of individual cells while retaining the ability to locate the cell within a food tissue system. This paper reviews the application of confocal microscopy in food research and direct observation of viable bacteria with emphasis on their use in food microbiology.

Bacteria↗

Nordic dentists' opinions on the safety of amalgam and other dental restorative materials.

The safety of amalgam and other restorative materials has caused concern among dental patients in recent years. The aim of this study was to obtain information on dentists' perceived competence in handling different filling materials and their opinions on the safety of these. A random sample of practising dentists in Denmark, Finland, Norway, and Sweden received a mail questionnaire in spring 1990. Answers were received from 1732 dentists (65%). The study showed that the respondents believed that their theoretic knowledge and clinical skills were generally at a high level regarding restorative materials. The risks of the side-effects of gold, ceramic materials, and glass ionomer were considered to be low by about 90% of the respondents. Amalgam was considered to be significantly more hazardous by the Swedish respondents than the others. Interestingly, composite was considered to be associated with a high risk of side-effects by about half of the dentists in all Nordic countries. The dentists' opinions were not found to be greatly influenced by their sex, age, or place of residence but rather by their country and service sector. Against the background of the present lack of scientific evidence on the hazardousness of amalgam or other restorative materials for patients' general health, these findings indicate that dentists are influenced by discussions in the mass media about dental treatment and materials and, of course, by the guidelines given by the health authorities in their own countries. Few dentists were shown to be concerned about occupational risks associated with the use of amalgam, and they had not had their own amalgam fillings replaced.(ABSTRACT TRUNCATED AT 250 WORDS)

Attitude of Health Personnel↗

Development of a laboratory robotic system for automated bioanalytical methods--II. A robot computer program for guarding totally automated bioanalytical methods.

The application of fully automated, unattended sample preparation performed by a laboratory robot for the analysis of drugs in biological samples requires the prevention of system failures which may arise in the on-line coupled chromatographic system or in other components of the robotic system. A computer program has been developed which can help to detect such problems. The control program for the robotic sample preparation contains a number of safety measures to intercept robotic or human errors. A routine is implemented, guarding for chromatographic malfunctions and errors in dispensing liquids by the robot. After detection of trouble, sample preparation is interrupted.

Algorithms↗

Physician concerns about vaccine adverse effects and potential litigation.

OBJECTIVE: To understand physician concerns about litigation and beliefs regarding vaccine safety. DESIGN: A stratified random sample of family physicians, pediatricians, and general practitioners younger than 65 years who were in office-based practices across the United States was selected from the American Medical Association list that includes nonmembers. A standardized telephone survey was conducted by trained interviewers in 1995. PARTICIPANTS: Physicians seeing 5 or more patients per week younger than 6 years and having 50% or more primary care patients were eligible for the study. RESULTS: Of the 1236 physicians who were surveyed, 32% and 13% overestimated the risk for serious adverse effects related to pertussis and measles vaccines, respectively. Among physicians who thought that serious adverse effects from diphtheria and tetanus toxoids and pertussis vaccine (DTP) were unlikely, 15% were highly concerned about litigation; however, among those with higher ratings of the likelihood of serious adverse effects, 38% were highly concerned about vaccine litigation (P < .01). Of those aware of the Vaccine Injury Compensation Program, only 41% believed that it afforded a high level of liability protection; 22% believed that it gives little protection, and 37% gave an intermediate answer. Among physicians highly concerned about vaccine litigation, 22% were unlikely to recommend the third dose of DTP for a child with a fever of 39.4 degrees C and no other symptoms after the second dose of DTP, whereas among those expressing little concern about litigation, only 12% were unlikely to vaccinate (P < .05). Although some physicians were concerned about litigation, most (86%) encouraged vaccination even if a parent was argumentative about possible adverse effects. CONCLUSION: Physicians' perceptions about the risk for adverse effects and protection afforded by the Vaccine Injury Compensation Program influence their concern about litigation and, to a lesser extent, their reported likelihood to administer immunizations.

Adult↗

Evaluation of a safety resheathable winged steel needle for prevention of percutaneous injuries associated with intravascular-access procedures among healthcare workers.

OBJECTIVE: To compare the percutaneous injury rate associated with a standard versus a safety resheathable winged steel (butterfly) needle. DESIGN: Before-after trial of winged steel needle injuries during a 33-month period (19-month baseline, 3-month training, and 11-month study intervention), followed by a 31-month poststudy period. SETTING: A 1,190-bed acute care referral hospital with inpatient and outpatient services in New York City. PARTICIPANTS: All healthcare workers performing intravascular-access procedures with winged steel needles. INTERVENTION: Safety resheathable winged steel needle. RESULTS: The injury rate associated with winged steel needles declined from 13.41 to 6.41 per 100,000 (relative risk [RR], 0.48; 95% confidence interval [CI95], 0.31 to 0.73) following implementation of the safety device. Injuries occurring during or after disposal were reduced most substantially (RR, 0.15; CI95, 0.06 to 0.43). Safety winged steel needle injuries occurred most often before activation of the safety mechanism was appropriate (39%); 32% were due to the user choosing not to activate the device, 21% occurred during activation, and 4% were due to improper activation. Preference for the safety winged steel needle over the standard device was 63%. The safety feature was activated in 83% of the samples examined during audits of disposal containers. Following completion of the study, the safety winged steel needle injury rate (7.29 per 100,000) did not differ significantly from the winged steel needle injury rate during the study period. CONCLUSION: Implementation of a safety resheathable winged steel needle substantially reduced injuries among healthcare workers performing vascular-access procedures. The residual risk of injury associated with this device can be reduced further with increased compliance with proper activation procedures.

Blood-Borne Pathogens↗

Microbiological hazard identification and exposure assessment of street food vending in Johannesburg, South Africa.

One hundred and thirty-two samples of beef, chicken, salad and gravy were collected from two street vendors over eleven replicate surveys to assess microbiological safety and quality. For each food type samples were collected during preparation and holding. Dish water was also collected and food preparation surfaces swabbed during preparation and display. Standard methods were used to determine aerobic plate counts, Enterobacteriaceae counts, coliform counts and spore counts. Six hundred and seventy-five predominant colonies were isolated from aerobic plate counts of all samples and characterised. The incidence of selected foodborne bacterial pathogens and non-pathogenic E. coli 1 was also determined. In most cases mean bacterial counts of the raw materials were significantly higher (P < 0.05) than those of corresponding cooked foods. No significant differences (P > 0.05) in all count types were observed between food samples collected during cooking and those collected during holding. In addition, no significant differences (P > 0.05) in all count types were observed between prepared salads and their raw materials. Mean bacterial counts of water and swab samples collected from vendor 1 were lower than those of water and swab samples collected from vendor 2.The predominant populations isolated from the aerobic plate counts were Bacillus spp., Staphylococcus spp., Enterobacteriaceae and Alcaligenes spp. Bacillus cereus was detected in 17%, Clostridium perfringens in 1%, Staphylococcus aureus in 3% and Vibrio metchnikovii in 2% of the food samples. Campylobacter jejuni, Listeria monocytogenes, and Escherichia coli O157:H7 were not detected. Non-pathogenic E. coli 1 was detected in 13% of food samples, in 86 and 36% of dish water samples collected from vendors 1 and 2, respectively, and in 36% of surface swab samples from vendor 2.

Animals↗

Results of a survey of small-animal practitioners on the perceived clinical efficacy and safety of an oral nutraceutical.

A mail survey of 3080 practitioners from two sampling frames was used to determine the perceived clinical efficacy and safety of an oral nutraceutical for the treatment of degenerative joint disease (DJD) in small animals. Overall response rate was 82%, of which 64% reported that they were recommending the oral nutraceutical to their clients. There was a high variability reported between the severity of DJD and response to treatment. Sixty-eight percent of practitioners reported lame dogs older than five years to be the most responsive to treatment. Most practitioners (83%) believed response to treatment with the studied product occurred within four weeks. Practitioners using the product rated the clinical efficacy of the product to be either 'good' or 'excellent' in improving mobility, alleviating pain and improving attitude in the majority of the treated animals. No practitioner reported a worsening of pain, mobility or attitude. Self-reported adverse effects were 2% (the most common was gastrointestinal upset). The results of this survey indicated that perceived clinical efficacy of the studied nutraceutical in the treatment of DJD was regarded to be 'good' or 'excellent' by most of the practitioners who use this product and was considered to be safe with minimal side effects. This provides additional information on the product for use in planning future clinical trials.

Administration, Oral↗

Quantitative determination of CGS 18102A, a new anxiolytic, in human plasma using capillary gas chromatography/mass spectrometry.

CGS 18102A is a novel hexahydrobenzopyranopyridine that has a mixed pharmacological profile of 5-HT-1A agonist and 5-HT-2 antagonist properties. Based upon these mechanisms, the compound is predicted to have anxiolytic efficacy with possible efficacy in depression. Preclinical studies in the rat have shown the drug to be well absorbed and extensively metabolized. Because of the anticipated low plasma levels in humans a gas chromatography/mass spectrometry (GC/MS) analytical method has been developed and validated to determine plasma concentrations of CGS 18102A in early clinical studies. The method utilizes CGS 18416A as the internal standard. Samples (1 mL) were extracted with pentane:ethyl acetate (75:25, v:v). Extracts were then concentrated and analysed directly by GC/MS. Separation was accomplished on a methylsilicone capillary column (30 m x 0.32 mm i.d.). GC/MS was carried out under positive ion ammonia CI conditions, with selected ion monitoring of the [M + H]+ ions (m/z = 262 and 248) for CGS 18102A and CGS 18416A, respectively. The method was successively applied to the analysis of clinical samples from an ascending multidose safety and tolerability study conducted in six normal healthy male volunteers.

Adolescent↗

Lesbians' and gay men's experiences of discrimination and harassment in a university community.

A survey of 125 lesbians and gay men in a university community was conducted to determine the incidence of discrimination, harassment, and violence. Nearly three fourths had experienced verbal abuse; 26% were threatened with violence; and 17% had personal property damaged. Students and roommates were most often those responsible. Most incidents were not reported to authorities, and many made changes in their daily routines to avoid harm. Over half of the sample feared for their personal safety; their fear was related to the amount of harassment and previous property damage. Men were more often victimized than women.

Adolescent↗

Exposure to airborne asbestos in buildings.

The concentration of airborne asbestos in buildings and its implication for the health of building occupants is a major public health issue. A total of 2892 air samples from 315 public, commercial, residential, school, and university buildings has been analyzed by transmission electron microscopy. The buildings that were surveyed were the subject of litigation related to suits alleging the general building occupants were exposed to a potential health hazard as a result of exposure to the presence of asbestos containing materials (ACM). The average concentration of all asbestos structures was 0.02 structures/ml (s/ml) and the average concentration of asbestos greater than or equal to 5 microns long was 0.00013 fibers/ml (f/ml). The concentration of asbestos was higher in schools than in other buildings. In 48% of indoor samples and 75% of outdoor samples, no asbestos fibers were detected. The observed airborne concentration in 74% of the indoor samples and 96% of the outdoor samples is below the Asbestos Hazard Emergency Response Act clearance level of 0.01 s/ml. Finally, using those fibers which could be seen optically, all indoor samples and all outdoor samples are below the Occupational Safety and Health Administration permissible exposure level of 0.1 f/ml for fibers greater than or equal to 5 microns in length. These results provide substantive verification of the findings of the U.S. Environmental Protection Agency public building study which found very low ambient concentrations of asbestos fibers in buildings with ACM, irrespective of the condition of the material in the buildings.

Air Pollutants↗

Lateral automobile impacts and the risk of traumatic brain injury.

STUDY OBJECTIVES: We determine the relative risk and severity of traumatic brain injury among occupants of lateral impacts compared with occupants of nonlateral impacts. METHODS: This was a secondary analysis of the National Highway Traffic Safety Administration's National Automotive Sampling System, Crashworthiness Data Systems for 2000. Analysis was restricted to occupants of vehicles in which at least 1 person experienced an injury with Abbreviated Injury Scale score greater than 2. Traumatic brain injury was defined as an injury to the head or skull with an Abbreviated Injury Scale score greater than 2. Outcomes were analyzed using the chi2 test and multivariate logistic regression, with adjustment of variance to account for weighted probability sampling. RESULTS: Of the 1,115 occupants available for analysis, impact direction was lateral for 230 (18.42%) occupants and nonlateral for 885 (81.58%) occupants. One hundred eighty-seven (16.07%) occupants experienced a traumatic brain injury, 14.63% after lateral and 16.39% after nonlateral impact. The unadjusted relative risk of traumatic brain injury after lateral impact was 0.89 (95% confidence interval [CI] 0.51 to 1.56). After adjusting for several important crash-related variables, the relative risk of traumatic brain injury was 2.60 (95% CI 1.1 to 6.0). Traumatic brain injuries were more severe after lateral impact according to Abbreviated Injury Scale and Glasgow Coma Scale scores. The proportion of fatal or critical crash-related traumatic brain injuries attributable to lateral impact was 23.5%. CONCLUSION: Lateral impact is an important independent risk factor for the development of traumatic brain injury after a serious motor vehicle crash. Traumatic brain injuries incurred after lateral impact are more severe than those resulting from nonlateral impact. Vehicle modifications that increase head protection could reduce crash-related severe traumatic brain injuries by up to 61% and prevent up to 2,230 fatal or critical traumatic brain injuries each year in the United States.

Abbreviated Injury Scale↗

Quetiapine for the treatment of borderline personality disorder; an open-label study.

PURPOSE: Only a few studies have commented on the use of atypical antipsychotics for the treatment of Borderline Personality Disorder (BPD) features, including affective dysregulation and aggression. We aimed at evaluating both efficacy and safety of quetiapine in a sample of consecutive BPD patients. GENERAL METHODS: 29 BPD outpatients entered, and 23 completed, a 12 week, open-label, regime of quetiapine at an average daily dosage of 540 mg (range: 400-800 mg). Efficacy assessment psychometric instruments included: Hamilton Rating Scales for Depression (HAM-D); Brief Psychiatric Rating Scale (BPRS); Global Assessment of Functioning (GAF); Clinical Global Impression Scale (CGI); and Aggression Questionnaire (AQ). FINDINGS: Both completer and intent-to-treat analysis showed that most psychometric scales' scores exhibited a highly significant (HAM-D: p=.003; BPRS Hostility and Suspiciousness subscales; CGI; GAF; AQ: all at p=.000) improvement over time. Six patients dropped out early from treatment due to side effects; quetiapine was associated with two cases of transient thrombocytopenia. CONCLUSIONS: Present findings would suggest that quetiapine may be effective for the treatment of a number of BPD features, including low mood and aggression. However, monitoring blood counts in patients receiving quetiapine seems to be justified.

Adult↗

DNA extraction and Escherichia coli quantification of anaerobically digested biosolids using the competitive touchdown PCR method.

Accurate enumeration of indicator organisms such as Escherichia coli is important for assessing the safety of water and wastewater samples. Recent research has shown that E. coli can enter a viable but non-culturable state; therefore, traditional cultivation methods could potentially underestimate the quantities of the organisms. The goals of the research were to develop and verify a DNA extraction protocol and a quantitative polymerase chained reaction (PCR) method for E. coli enumeration in digested biosolids. A solvent-based DNA extraction protocol with extensive cell lysis recovered approximately 78-84% of spiked DNA. In comparison, a commercial kit only recovered 28-42% of DNA, likely from inefficient cell lysis. The developed competitive touchdown PCR (cPCR) method for E. coli enumeration was comparable to both real-time PCR (rt-PCR) and cultivation methods with sensitivity of approximately 50,000-500,000 E. coli per gram dry solids (DS), which is suitable for Class B biosolids monitoring in the US and "conventional" biosolids in the European Union. The cPCR protocol provides a less expensive alternative than the rt-PCR as a culturing independent method for enumerating E. coli.

Anaerobiosis↗

Immediate and reversible platelet inhibition after intravenous administration of a peptide glycoprotein IIb/IIIa inhibitor during percutaneous coronary intervention.

We studied the pharmacokinetic and pharmacodynamic properties of integrelin, a novel platelet glycoprotein IIb/IIIa receptor inhibitor, in patients undergoing elective percutaneous coronary intervention. Patients were randomized to placebo (n = 19) or to 1 of 4 integrelin dosing regimens (total n = 54) that were studied sequentially. All patients received aspirin and heparin. Patients were followed until discharge for the occurrence of adverse clinical events: death, myocardial infarction, coronary artery bypass surgery, repeat intervention, or recurrent ischemia. Bleeding was the primary safety end point. Frequent blood sampling was performed for adenosine diphosphate-induced platelet aggregations. Simplate bleeding times were performed. Adverse clinical events occurred less often in the integrelin-treated patients, although the overall numbers were too small to make a definitive statement as to clinical efficacy. There was no significant increase in serious bleeding among integrelin-treated patients. The 2 highest integrelin boluses (180 and 135 micrograms/kg) immediately (15 minutes after the bolus) provided > 80% inhibition of adenosine diphosphate-induced platelet aggregation in > 75% of treated patients. A constant integrelin infusion of 0.75 micrograms/kg/min maintained this marked antiplatelet effect, whereas an infusion of 0.50 micrograms/kg/min allowed gradual recovery of platelet function. Elective coronary intervention was performed safely and with no significant increase in serious bleeding events using integrelin with aspirin and heparin as an antithrombotic regimen. Integrelin provided rapid, intense, and persistent ex vivo platelet inhibition during coronary intervention. This new antiplatelet agent may be beneficial in reducing platelet-mediated ischemic complications of percutaneous coronary intervention.

Aged↗

Early feeding after cesarean: randomized trial.

OBJECTIVE: To study the rate of ileus symptoms and hospital course of women who are offered solid food shortly after cesarean delivery. METHODS: This study involved women delivered by cesarean under regional anesthesia. Exclusion criteria included general anesthesia, magnesium sulfate, intra-operative bowel injury or bowel surgery, or other conditions that precluded early feeding. Early-fed women were offered regular diets within 8 hours of surgery, and controls were given nothing by mouth for 12-24 hours, advanced to clear liquids on the first postoperative day, and then given solid food on the second or third postoperative day. RESULTS: Sixty women were assigned randomly to each method. Early-fed women received solid food sooner after surgery, 5.0 +/- 1.2 hours versus 40.0 +/- 10.6 hours. The incidences of mild ileus symptoms and postoperative complications were similar in both groups; however, the study did not have an adequate sample size to definitively assess safety concerns. Women in the early-fed group had shorter hospital stays (49.5 +/- 12.7 hours versus 75.0 +/- 12.3 hours, P <.001), and shorter time intervals from surgery to bowel movement, 34.5 hours (interquartile range 25.3-48.8) versus 51.0 (43.3-62.0) hours, P <.001. In the early-fed group, women whose operative times exceeded 40 minutes were more likely to have symptoms of mild ileus. CONCLUSION: Early initiation of solid food after cesarean delivery appears to be well tolerated and may be associated with a shorter hospital stay. Early-fed women whose operations exceed 40 minutes may be more likely to have mild ileus symptoms.

Adult↗

Pooling of samples for seroepidemiological surveillance of human T-cell lymphotropic virus types I and II.

We evaluated a straight forward pooling strategy for antibody screening of HTLV-I/II, using panels of sera from various parts of the world including a total of 43 HTLV-I and 54 HTLV-II positive specimens. Four antibody screening assays were included in the evaluation: the HTLV-I/II GE 80/81 (Murex Diagnostics), the HTLV-I/HTLV-II Ab Capture ELISA (Ortho Diagnostics), the HTLV-I/II ELISA 3.0 (Genelabs Diagnostics) and the Serodia HTLV-I (Fujirebio). The Murex and Ortho assays represent a new generation of HTLV screening tests with a sandwich format incorporating both HTLV-I and HTLV-II synthetic and/or recombinant peptide antigens. The Genelabs assay is an indirect ELISA with recombinant HTLV-I and -II antigens and Serodia is a particle agglutination assay with HTLV-I whole viral lysate. Each HTLV-positive sample was included in pools of 1/1 up to 1/16, in two-fold steps made in normal HTLV-negative blood donor serum from one up to nine donors. For HTLV-I, with the exception of one false negative sample in dilution 1/16 with Genelabs ELISA, all assays were positive at all dilutions. The Murex assay had absorbance values at maximum levels for all samples at all dilutions. The other assays had gradually decreasing absorbance values although clearly above cut-off. For HTLV-II, the Murex assay correctly detected all samples to dilution 1/16 despite gradually decreasing signals. The Serodia assay had 100% sensitivity to dilution 1/4 while at 1/8 and 1/16 it decreased 82 and 80%, respectively. The Genelabs ELISA had gradually decreasing sensitivity for HTLV-II from 98 (1/1) to 33% (1/16) while the Ortho assay detected all specimens at all dilutions in a limited set of samples tested. Taken together, this evaluation has shown that pooling of samples may be an appropriate strategy for serosurveillance of HTLV. It is, however, crucial to limit the number of samples and to choose assays that allow the dilution caused by the pooling. Using the best performing assays in this evaluation for pools of e.g. five samples would leave a reasonable safety margin.

Agglutination Tests↗

Determination of a potential antiasthmatic compound, tibenelast, and its application to phase I clinical trials.

A sensitive and selective high-performance liquid chromatographic (HPLC) assay was developed for the determination of tibenelast, 5,6-diethoxybenzo[b]thiophene-2- carboxylic acid, in plasma and urine. The plasma assay involves protein precipitation with 4% trichloroacetic acid, while the urine assay is an automated solid-phase extraction procedure that utilizes the Waters Millilab workstation. The analysis was achieved by reversed-phase HPLC with ultraviolet detection at 313 nm. The quantitation limit of the assay was 50 ng/ml in plasma and 100 ng/ml in urine. The intra-day coefficient of variation for the plasma analysis was between 2.2 and 8.4%, while the overall inter-day coefficient of variation was 5.5 and 6.0% for the high and low calibration curves, respectively. The intra-day coefficient of variation for the urine analysis was between 0.3 and 3.0%, while the inter-day coefficient of variation was 2.1% for both the low and high validation samples. The assay methodology has been used in the evaluation of samples from pharmacokinetic and clinical safety studies.

Asthma↗

"On a supposed right to lie [to the public] from benevolent motives": communicating health risks to the public.

There are three main categories of rationale for withholding information or telling lies: if overwhelming harm can only be averted through deceit; complete triviality such that it is irrelevant whether the truth is told; a duty to protect the interests of others. Public health authorities are frequently having to form judgements about the public interest, whether to release information or issue warnings. In June 1992, routine surveillance detected patulin levels (a known carcinogen) in samples of apple juice exceeding safety threshold. Remedial actions were promptly taken and it was planned to subsequently publish the information in the routine way. However, the media portrayed the handling of the problem as a conspiracy and there was a short term reduction in juice sales. In October 1995, the UK Committee on Safety of Medicines issued a warning about certain brands of the contraceptive pill, based on the interim results of three unpublished studies. The increased risk of thromboembolism was small, but the resulting scare led to an increase in unwanted pregnancies. The handling of the B.S.E. crisis in the U.K. also led to accusations of incompetence or conspiracy. Public health authorities have to handle uncertainty and frequently have to form judgements for public safety on the basis of evidence of poor quantity and quality. Their task is not helped by the sometimes conflicting agenda of scientists and media. The public also have differing perceptions and interpretations of risk. The series of scares and crises are having a detrimental effect on public confidence in public health authorities.

Animals↗