Comparison of specimen collection and laboratory techniques for isolation of Haemophilus ducreyi.

Sixteen patients with clinical chancroid were studied prospectively; different culture media and sampling techniques from genital lesions were evaluated. Technique A was aspiration of a saline wash from the ulcer which was pooled and inoculated into rabbit blood, rabbit blood + vancomycin (5 microgram/ml), and semisolid chocolate agar + vancomycin (3 microgram/ml). Each primary culture medium was subcultured to chocolate agar with 1% IsoVitaleX (CA), CA with vancomycin (3 microgram/ml) plus polymyxin (7.5 microgram/ml; CA + vp). Technique B was the use of a cotton swab, plated directly on CA, CA + v, and CA + vp. Nine strains of Haemophilus ducreyi were obtained. Technique A yielded seven strains, whereas technique B yielded eight strains; with each technique, five strains were isolated only after use of selective antibiotic media. CA + v medium yielded the largest number of isolates. Direct inoculation by swab to CA + v from chancroidal ulcers is effective as an isolation technique for growth of H. ducreyi.     

Retrospective data collection and analytical techniques for patient safety studies

To enhance patient safety, data about actual clinical events must be collected and scrutinized. This paper has two purposes. First, it provides an overview of some of the methods available to collect and analyze retrospective data about medical errors, near misses, and other relevant patient safety events. Second, it introduces a methodological approach that focuses on non-routine events (NRE), defined as all events that deviate from optimal clinical care. In intermittent in-person surveys of anesthesia providers, 75 of 277 (27%) recently completed anesthetic cases contained a non-routine event (98 total NRE). Forty-six of the cases (17%) had patient impact while only 20 (7%) led to patient injury. In contrast, in the same hospitals over a two-year period, we collected event data on 135 cases identified with traditional quality improvement processes (event incidence of 0.7-2.7%). In these quality improvement cases, 120 (89%) had patient impact and 74 (55%) led to patient injury. Preliminary analyses not only illustrate some of the analytical methods applicable to safety data but also provide insight into the potential value of the non-routine event approach for the early detection of risks to patient safety before serious patient harm occurs.     

Genetic quality control of laboratory animals used in aging studies

Laboratory rodents provide a useful model for aging processes in humans. Various genetic “types” of stock are available, including outbred stocks and inbred strains and their derivatives. Inbred strains, which can be regarded as clones of genetically identical individuals, provide a powerful research tool for studies in many disciplines, including gerontology. However, some form of genetic quality control is essential to ensure that the strains are authentic. Single gene polymorphisms, particularly those detected by electrophoresis and immunological methods, provide a powerful tool for such quality control, though these methods are expensive and require considerable expertise. Methods based on several loci studied simultaneously include skin grafting and polyvalent alloantisera. These methods are often quick and technically relatively easy, but are less flexible than the single locus methods. Methods based on DNA restriction fragment length polymorphisms (RFLPs) fall into both categories depending on whether a single locus or a multilocus probe, such as the fingerprinting probes, is used. These DNA-based methods have many advantages, and are likely to be the methods of choice in the future.     

Mycoplasma iowae: field and laboratory studies to evaluate egg transmission in turkeys

In order to study the egg transmission of Mycoplasma iowae in turkeys, two experiments are undertaken. Experiment 1 was a field trial designed to investigate egg transmission in naturally infected turkeys: commercial hens were inseminated with semen from naturally infected toms. Mycoplasma iowae was regularly cultured from dead-in-shell embryos, pips and culls but not from healthy poults. Egg transmission occurred from the second up to the tenth week of production. Experiment 2 was a laboratory study conducted to observe persistence and egg transmission of M. iowae in experimentally infected turkeys. Specific pathogen free (SPF) hens were inoculated either intracheally or intravaginally. They were then artificially inseminated with semen from SPF toms. Very little egg transmission was observed. The cultures from tracheal or vaginal swabs of the inoculated hens showed a limited invasive ability of M. iowae in the adult bird. The humoral immune response as detected by the metabolism inhibition test, was very poor. The observations made in these two experiments show the importance of venereal spread of M. iowae, especially under artificial insemination. Consequently, the need for monitoring toms is obvious.     

Measurement and interpretation issues in laboratory and field studies of energy expenditure

The methods for measuring energy expenditure in the laboratory and the field are described and critically appraised and the criteria for reliable and valid measurements identified. Variation and adaptation in the energy costs of activities and the total daily energy expenditure are considered in the context of discriminating between what is variation and adaptation and what is honest error. This involves issues of comparative energetics and standardization and oxygen uptake kinetics, which are frequently neglected or ignored. The energetics of physical work, where many of the problems arise, are examined. It is technically feasible to measure energy expenditure with high accuracy, but the methods required, calorimeter rooms and doubly labelled water, are not widely available or applicable. The field methods, the factorial method of diary record of activities and their energy costs, and the heart rate methods are of acceptable accuracy for groups but less so for individuals. When using energy costs from the literature, as has been common in human biology, several problems arise. The sample sizes are small and of unknown provenance, and there is no information on variability or on whether the values are based on continuous or intermittent work. The problem of how to allow for differences in size and composition has been tackled with convenience rather than concepts. It is recommended that isometric scaling, using kcal/kg, be avoided and allometric scaling by, e.g., analysis of covariance, be used to remove the effects of size and composition. In investigations of variation and adaptation in the efficiency of work, the effects of oxygen uptake kinetics and of anaerobic metabolism must be considered. © 1996 Wiley-Liss, Inc.     

Qualitative assessment of study materials and communication strategies used in studies that include DNA collection

To understand motivations and barriers to participation in studies that include DNA collection, focus group discussions were held with mothers who had participated in a case-control study of birth defects. Recruited mothers had completed an interview and had received a mailed kit containing cytobrushes to collect buccal cells for DNA from herself, her infant, and her infant's father. Six moderator-led focus groups were attended by a total of 38 women residing in Atlanta, Georgia. Focus groups were segmented by DNA collection status (biologics participants or nonparticipants), infant case-control status, infant birthweight, and maternal race and ethnicity. This report assesses maternal attitudes toward study materials and communication strategies. Across groups, respondents expressed concern about how their contact information was obtained. Study materials were described as clear and professional by most women, although some respondents reported confusion about disclosure of individual genetic results. Respondents generally reported that monetary incentives were not a motivation to participate, but increased perceived study legitimacy. Biologics nonparticipants expressed concerns about kit component sterility; government involvement; and DNA sample use, storage, and disposal. Respondents suggested that investigators provide feedback on whether sample collection was performed correctly and provide materials targeted to fathers to help alleviate paternal skepticism. Participation in DNA collection might be improved by strengthening study materials and communication strategies.     

Palliative care making a difference in rural Uganda,Kenya and Malawi: three rapid evaluation field studies

Background

Many people live and die in pain in Africa. We set out to describe patient, family and local community perspectives on the impact of three community based palliative care interventions in sub-Saharan Africa.

Methods

Three palliative care programmes in Uganda, Kenya and Malawi were studied using rapid evaluation field techniques in each country, triangulating data from three sources: interviewswith key informants, observationsof clinical encounters and the local health and social care context, and routine data from local reports and statistics.

Results

We interviewed 33 patients with advanced illness, 27 family carers, 36 staff, 25 volunteers, and 29 community leaders and observed clinical care of 12 patients. In each site, oral morphine was being used effectively. Patients valued being treated with dignity and respect. Being supported at home reduced physical, emotional and financial burden of travel to, and care at health facilities. Practical support and instruction in feeding and bathing patients facilitated good deaths at home.In each country mobile phones enabled rapid access to clinical and social support networks. Staff and volunteers generally reported that caring for the dying in the face of poverty was stressful, but also rewarding, with resilience fostered by having effective analgesia, and community support networks.

Conclusions

Programmes were reported to be successful because they integrated symptom control with practical and emotional care, education, and spiritual care. Holistic palliative care can be delivered effectively in the face of poverty, but a public health approach is needed to ensure equitable provision.

     

Evaluation of quantitative aerosol techniques for use in bronchoprovocation studies

To investigate airway physiology by use of inhaled aerosols, it is frequently necessary to measure the actual amount of material deposited on the airway wall as well as the site of particle deposition. To satisfy these needs, radiolabeled aerosols and gamma camera techniques have been used to measure regional deposition of inhaled particles. To make quantitative measurements of the amount deposited, previous investigators have used a "phantom" technique to indirectly calibrate the gamma camera for the attenuation of gamma rays through the lungs and chest wall. For this calibration, the phantom is a simulated lung containing a known amount of radioactivity. Radioactive counts emitted from the phantom are assumed to be attenuated in the same manner as the intact human lung. The present article describes a technique to determine directly the amount of inhaled aerosol deposited in the lung and simultaneously to calibrate the gamma camera for each individual subject. We used right angle light scattering and a gamma camera to measure individual values of the deposition fraction (DF) of inhaled aerosol deposited in the lung and the coefficient of attenuation (AC) of gamma rays in normal and obstructed lungs of human subjects. Radiolabeled monodisperse aerosols 1 and 2 microns in diameter were used. Knowledge of the activity of the inhaled aerosol (microcurie per liter), the volume inhaled, and the measured DF determined each subject's AC (counts per minute per microcurie). DF varied by an order of magnitude in normal (0.04 to 0.48) and obstructed (0.16 to 0.75) of subjects.(ABSTRACT TRUNCATED AT 250 WORDS)     

Epicillin: in vitro laboratory studies

A new semisynthetic penicillin, structurally related to ampicillin, has been assigned the generic name epicillin, 6-[d-2-amino-2-(1, 4-cyclohexadienyl) acetamido]-penicillanic acid. The antimicrobial spectrum and level of activity of epicillin in vitro are similar to those of ampicillin. In studies with recent clinical isolates, these two antibiotics, when compared with carbenicillin, showed consistently higher antimicrobial activity against Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, and Proteus species. When tested against Pseudomonas aeruginosa isolates, on the other hand, epicillin exhibited a level of intrinsic activity superior to that of ampicillin but less than that of carbenicillin. Epicillin is an amphoteric substance that is sensitive to penicillinase, is acid-stable, and is minimally, but reversibly, bound to human serum protein.     

Trimethoprim: laboratory and clinical studies

Trimethoprim is a synthetic compound which arrests the step in bacterial purine synthesis immediately succeeding that interfered with by sulphonamides.Acting alone it inhibits the growth of a wide range of bacteria in generally much lower concentrations than sulphonamides. Its action with a sulphonamide is strongly synergic and bactericidal.Studies are described of suitable cultural conditions, methods of testing sensitivity, and acquired resistance.Some clinical results of its use are reported in respiratory tract infections, Gram-negative septicaemia, and urinary tract infections, and further therapeutic possibilities are discussed.     

Blood collection procedures in hematology: knowledge and practice among laboratory personnel

Blood collection is an important preanalytical component of haematological testing. This questionnaire based study was conducted on laboratory personnel to elicit their knowledge and practice regarding blood collection procedures for haematological testing. Questionnaire comprised of 37 multiple choice questions, of which 10 questions each were related to essential and desirable knowledge, 10 to practice and 7 to educational and job profile. Ninety four laboratory personnel participated in the study. Analysis was done on SPSS software. Percentages of unsatisfactory scores were 42.6%, 4.3%, 17% and 6% in the essential knowledge, desirable knowledge, practice and total scoring respectively. 91.5%, however, had a satisfactory total score. Importance needs to be given to blood collection procedures both, individually and collectively. The study highlights the deficient areas which need to addressed by all laboratorians. Such studies should also be conducted among nursing and resident staff- the other groups concerned with blood collection.     

Evaluation of rodent bait containing imidacloprid for the control of fleas on commensal rodents in a plague-endemic region of northwest Uganda

In recent decades, the majority of human plague cases (caused by Yersinia pestis) have been reported from Africa. In an effort to reduce the risk of the disease in this area, we evaluated the efficacy of a host-targeted rodent bait containing the insecticide imidacloprid for controlling fleas on house-dwelling commensal rodents in a plague-endemic region of northwestern Uganda. Results demonstrated that the use of a palatable, rodent-targeted, wax-based bait cube was effective at reducing the prevalence of fleas on commensal rodents and flea burdens on these animals at day 7 postbait exposure, but lacked significant residual activity, allowing flea populations to rebound in the absence of additional bait applications. Our results indicate the use of a palatable host-targeted bait block containing imidacloprid was an effective technique for quickly reducing flea numbers on rodents in northwest Uganda and, thus, could be useful for lowering the potential risk of human flea bite exposures during plague outbreaks if applied continuously during the period of risk.     

Evaluation of various methods of maternal placental blood collection for immunology studies

The collection of maternal placental intervillous blood (IVB), without contamination of fetal blood and with an accurate mononuclear cell profile, is essential for immunological studies of placental malaria and other infectious diseases of the placenta. We have compared five documented methods of IVB collection: perfusion, incision, biopsy, tissue grinding, and puncture (prick) for fetal blood contamination and mononuclear cell profiles using flow cytometry. Twenty-five placentas were obtained from Plasmodium falciparum and human immunodeficiency virus-negative primigravid and secundigravid women delivering at Nyanza Provincial Hospital in Kisumu, western Kenya. Each of the five methods was performed on the same placenta. Fetal red blood cell contamination was significantly lower for the prick and perfusion methods (4.1% and 8.3%, respectively) than for incision (59.5%), biopsy (42.6%), and tissue grinding (19.9%). Significant variation was noted among the five methods in the percentages of monocytes, total T cells, CD4+ and CD8+ T cells, B cells, and NK cells. Further, a pairwise comparison of prick and perfusion, the two methods with low fetal blood contamination, showed that they were not different for fetal red blood cell contamination levels; however, prick yielded significantly higher percentages of CD4 T cells and CD4 memory T cells than perfusion. Collection by prick was determined to be the best method of intervillous blood collection for immunology studies, and perfusion represented the next best method of choice due to high sample volume yield. Overall, in considering the advantages/disadvantages of the two methods with low fetal cell contamination, we conclude that a combination of prick and perfusion is most suitable for immunology studies.     

Measuring arterial blood pressure in humans: Auscultatory and automatic measurement techniques for human biological field studies

Human biologists have been examining arterial blood pressure since they began studying the effects of the environment and culture on the health of diverse populations. The Korotkoff auscultatory technique with a trained observer and aneroid sphygmomanometer is the method of choice for blood pressure measurement in many bioanthropological field contexts. Korotkoff sounds (the first and fifth phases) are the preferred determinants of systolic and diastolic pressure, even in infants, children, pregnant women, and the elderly. Training of observers, positioning of individuals, and selection of cuff size are all essential for obtaining standardized measurements. Automatic electronic devices are increasingly being used for blood pressure measurement in human biological studies. The automatic monitors often use the oscillometric method for measuring pressure, but must be validated before use. The emergence of automatic ambulatory blood pressure monitors has opened another avenue of research on blood pressure in human biology, where allostasis and circadian responses to environmental change and real life behavioral challenges can be defined and evaluated, largely because there is now the ability to make multiple measurements over time and in varying contexts. Stand‐alone automatic monitors can also be substituted for manual auscultated readings in field contexts, although in studies where participants measure their own pressure, education about how the devices work and protocol specifics are necessary. Finally, computer‐driven plethysmographic devices that measure pressure in the finger are available to evaluate short‐term reactivity to specific challenges.     

Defined epitope blocking with Murray Valley encephalitis virus and monoclonal antibodies: laboratory and field studies.

In an attempt to develop a specific serological test for Murray Valley encephalitis (MVE) virus antibodies, a panel of MVE monoclonal antibodies was utilised in defined-epitope blocking ELISA tests. In sera of mice immunised singly and in combinations of MVE, Alfuy (ALF), and Kunjin (KUN) viruses, blocking patterns usually distinguished MVE infections from those of the other flaviviruses. When blocking tests with selected MAbs were applied to 468 flavivirus antibody positive sera collected from human subjects throughout New South Wales, sera with blocking patterns consistent with previous MVE infection were found in 18 subjects. All were long-term residents of areas previously frequented by MVE, and all were of an age to have been exposed to the virus in past epidemics. No such sera were found in subjects living in coastal areas of NSW where MVE has never been reported.     

Evaluation of physical fitness from field tests at high altitude in circumpubertal boys: comparison with laboratory data

Field tests of running and laboratory tests were performed in La Paz [high altitude (HA), 3700 m] and in Clermont-Ferrand [low altitude (LA), 300 m] to investigate their validity at HA. Prepubertal boys of mean ages 10.6 years (HA1,n = 16; LA1,n = 28) and pubertal boys of 13.7 years (HA2,n = 12; LA2,n = 41) took part in the study. All the boys performed a 30-m sprint (v _30m), a 30-s shuttle run (v _3os) and a progressive shuttle run test until their maximal aerobic velocity (v _maxsRT). Maximal oxygen consumption was extrapolated from the last test. . In the laboratory, the boys performed a force-velocity test (P _max), a Wingate test (P _Wing) and a graded test to measure maximal oxygen consumption ; direct method) on a cycle ergometer. At similar ages, there was no significant difference between HA and LA boys forv _30m andP _max. Thev _30s of HA boys was 3%–4% lower than those of LA boys (P<0.05); there was no significant difference forP _Wing. Significant relationships were observed at both altitudes betweenP _max (watts per kilogram) andv _30m (HA:r=0.76; LA:r=0.84) and betweenP _Wing andv _30s (HA:r=0.67; LA:r = 0.77); the slopes and the origins were the same at HA and LA. The ,v _maxSRT and were lower by 9%, 12% and 20%, respectively, at HA than at LA (P<0.05). However, the relationships between and (litres per minute) at HA (r=0.88) and at LA (r=0.93) were identical. In conclusion, chronic hypoxia did not modify performance in very short dash exercises. The influence of HA appeared when the exercise duration increased and, during a maximal shuttle run test, performance was reduced by 10% at HA. Moreover, it was possible to assessP _max,P _Wing and at HA as well as at LA from field tests.