Time to detection of the growth of Mycobacterium tuberculosis in MGIT 960 for determining the early bactericidal activity of antituberculosis agents.

Evaluation of early bactericidal activity (EBA) by the determination of a fall in viable colony-forming units (CFU) of Mycobacterium tuberculosis in sputum is a first step in the clinical study of new antituberculosis agents. The time to detection (TTD) of growth in liquid media is more sensitive and could substitute for CFU counting on solid media. Overnight sputum samples collected during the evaluation of the novel agent TMC207 in comparison to isoniazid and rifampicin were studied. For the determination of CFU, we incubated 10-fold dilutions of homogenized sputum on selective 7H10 agar. The TTD was measured by incubating decontaminated sputum in the BACTEC MGIT 960 system. The fall in bacillary load over 7 days determined by CFU counting closely matched the prolongation of the TTD in the BACTEC MGIT 960 system. The CFU counts correlated significantly with the TTD. While the ranking of agents and different dosages of TMC207 was similar, the highest dose of TMC207 showed markedly better activity when measured by the TTD than CFU counting when compared to the activity of isoniazid. Automated TTD could augment, or, in future, replace, CFU counting to determine sputum bacillary load in EBA clinical trials pending a more formal evaluation of the correlation of the measurements.

Locked down: Impact of COVID-19 restrictions on trauma presentations to the emergency department.

BACKGROUND: COVID-19 was recognised as a global pandemic on 11 March 2020. In South Africa (SA), a nationwide lockdown was implemented at midnight on 26 March to prepare for the predicted surge and slow the spread of the virus. OBJECTIVES: To compare the volume and type of presentations of trauma secondary to interpersonal violence and road traffic collisions (RTCs) during two 5-month periods, from February to June 2019 and 2020, in the emergency department (ED) of an academic tertiary hospital in Gauteng Province, SA. In 2020, February - June included the lockdown period. METHODS: An observational retrospective audit of the patient register at the Helen Joseph Hospital ED was conducted, comparing the number of trauma presentations secondary to interpersonal violence (assaults with gunshot wounds, general assaults including mob assaults, assaults with stab wounds) and RTC presentations between February and June 2019 and 2020. RESULTS: A total of 4 300 trauma presentations secondary to interpersonal violence and RTCs were noted in the 5-month period February - June 2019, as opposed to 3 239 presentations in February - June 2020 (25% decline). A 40% decline in the number of RTCs, from 1 704 in February - June 2019 to 1 026 in the corresponding period for 2020, was noted and found to be statistically significant (p=0.03). Declines in the volume of trauma cases secondary to interpersonal violence and of overall trauma cases were only directional in favour of 2020, but not statistically significant. CONCLUSIONS: The volume of trauma presentations secondary to interpersonal violence and RTCs in the Helen Joseph Hospital ED decreased during the lockdown period. The decline in the volume of RTCs was statistically significant, but declines in the volume of trauma presentations secondary to interpersonal violence and in the volume of overall trauma presentations were not.

Early bactericidal activity and pharmacokinetics of the diarylquinoline TMC207 in treatment of pulmonary tuberculosis.

Tibotec Medicinal Compound 207 (TMC207) is a novel diarylquinoline with a unique mode of action that targets mycobacterial ATP synthase. TMC207 exhibits high in vitro activity against mycobacterial strains either susceptible or resistant to all first-line and many second-line drugs, including fluoroquinolones, and has shown exceptional in vivo activity against several mycobacterial species in different animal models. In this early bactericidal activity study, 75 treatment-naïve patients with smear-positive pulmonary tuberculosis were randomized to once-daily oral TMC207 (25 mg, 100 mg, or 400 mg), 600 mg rifampin (RIF), or 300 mg isoniazid (INH) for 7 days. Sixteen-hour overnight sputum collected at baseline and on each treatment day was plated in serial dilutions on selective agar plates. The bactericidal activity was expressed as the log(10) decrease in CFU/ml sputum/day. Pharmacokinetic sampling was performed on day 7 of TMC207 administration up to 24 h postdose. The decreases in log(10) CFU counts (+/- standard deviation) from baseline to day 7 were 0.04 +/- 0.46 for 25 mg TMC207 (n = 14), 0.26 +/- 0.64 for 100 mg TMC207 (n = 14), 0.77 +/- 0.58 for 400 mg TMC207 (n = 14), 1.88 +/- 0.74 for INH (n = 11), and 1.70 +/- 0.71 for RIF (n = 14). Significant bactericidal activity of 400 mg TMC207 was observed from day 4 onward and was similar in magnitude to those of INH and RIF over the same period. The pharmacokinetics of TMC207 were linear across the dose range. In summary, TMC207 demonstrated bactericidal activity with a delayed onset and was well tolerated, and no study drug-related serious adverse events occurred.

Monitoring the ingestion of anti-tuberculosis drugs by simple non-invasive methods.

This investigation retrospectively assessed inexpensive non-invasive qualitative methods to monitor the ingestion of anti-tuberculosis drugs isoniazid, rifampicin and rifapentine. Results showed that commercial test strips detected the isoniazid metabolites isonicotinic acid and isonicotinylglycine as efficiently as the isonicotinic acid method in 150 urine samples. The presence of rifamycins in urine samples (n=1085) was detected by microbiological assay techniques and the sensitivity compared to the n-butanol extraction colour test in 91 of these specimens. The proportions detected by the two methods were significantly different and the sensitivity of the n-butanol procedure was only 63.8% (95% CL 51.2-76.4%) as compared to that of the superior microbiological method. Final validation (n=691) showed that qualitative assays measure isoniazid and rifamycin ingestion with an efficiency similar to high-performance liquid chromatography. The qualitative procedures may therefore be valuable in clinical trials and in tuberculosis clinics to confirm drug ingestion.

Radionuclides in fruit systems: model-model intercomparison study.

Modeling is widely used to predict radionuclide distribution following accidental radionuclide releases. Modeling is crucial in emergency response planning and risk communication, and understanding model uncertainty is important not only in conducting analysis consistent with current regulatory guidance, but also in gaining stakeholder and decision-maker trust in the process and confidence in the results. However, while methods for dealing with parameter uncertainty are fairly well developed, an adequate representation of uncertainties associated with models remains rare. This paper addresses uncertainty about a model's structure (i.e., the relevance of simplifying assumptions and mathematical equations) that is seldom addressed in practical applications of environmental modeling. The use of several alternative models to derive a range of model outputs or risks is probably the only available technique to assess consistency in model prediction. Since each independent model requires significant resources for development and calibration, multiple models are not generally applied to the same problem. This study uses results from one such model intercomparison conducted by the Fruits Working Group, which was created under the International Atomic Energy Agency (IAEA) BIOMASS (BIOsphere Modelling and ASSessment) Program. Model-model intercomparisons presented in this study were conducted by the working group for two different scenarios (acute or continuous deposition), one radionuclide ((137)Cs), and three fruit-bearing crops (strawberries, apples, and blackcurrants). The differences between models were as great as five orders of magnitude for short-term predictions following acute radionuclide deposition. For long-term predictions and for the continuous deposition scenario, the differences between models were about two orders of magnitude. The difference between strawberry, apple, and blackcurrant contamination predicted by one model is far less than the difference in prediction of contamination for a single plant species given by different models. This study illustrates the importance of problem formulation and implementation of an analytic-deliberative process in risk characterization.

Radionuclides in fruit systems: model prediction-experimental data intercomparison study.

This paper presents results from an international exercise undertaken to test model predictions against an independent data set for the transfer of radioactivity to fruit. Six models with various structures and complexity participated in this exercise. Predictions from these models were compared against independent experimental measurements on the transfer of 134Cs and 85Sr via leaf-to-fruit and soil-to-fruit in strawberry plants after an acute release. Foliar contamination was carried out through wet deposition on the plant at two different growing stages, anthesis and ripening, while soil contamination was effected at anthesis only. In the case of foliar contamination, predicted values are within the same order of magnitude as the measured values for both radionuclides, while in the case of soil contamination models tend to under-predict by up to three orders of magnitude for 134Cs, while differences for 85Sr are lower. Performance of models against experimental data is discussed together with the lessons learned from this exercise.

Radionuclide migration in forest ecosystems--results of a model validation study.

The primary objective of the IAEA's BIOMASS Forest Working Group (FWG) was to bring together experimental radioecologists and modellers to facilitate the exchange of information which could be used to improve our ability to understand and forecast radionuclide transfers within forests. This paper describes a blind model validation exercise which was conducted by the FWG to test nine models which members of the group had developed in response to the need to predict the fate of radiocaesium in forests in Europe after the Chernobyl accident. The outcomes and conclusions of this exercise are summarised. It was concluded that, as a group, the models are capable of providing an envelope of predictions which can be expected to enclose experimental data for radiocaesium contamination in forests over the time scale tested. However, the models are subject to varying degrees of conceptual uncertainty which gives rise to a very high degree of divergence between individual model predictions, particularly when forecasting edible mushroom contamination. Furthermore, the forecasting capability of the models over future decades currently remains untested.

Modelling and experimental studies on the transfer of radionuclides to fruit.

Although fruit is an important component of the diet, the extent to which it contributes to radiological exposure remains unclear, partially as a consequence of uncertainties in models and data used to assess transfer of radionuclides in the food chain. A Fruits Working Group operated as part of the IAEA BIOMASS (BIOsphere Modelling and ASSessment) programme from 1997 to 2000, with the aim of improving the robustness of the models that are used for radiological assessment. The Group completed a number of modelling and experimental activities including: (i) a review of experimental, field and modelling information on the transfer of radionuclides to fruit; (ii) discussion of recently completed or ongoing experimental studies; (iii) development of a database on the transfer of radionuclides to fruit; (iv) development of a conceptual model for fruit and (v) two model intercomparison studies and a model validation study. The Group achieved significant advances in understanding the processes involved in transfer of radionuclides to fruit. The work demonstrated that further experimental and modelling studies are required to ensure that the current generation of models can be applied to a wide range of scenarios.

Method of questioning and the accuracy of eyewitness testimony.

System variables are integrally part of factors that can be controlled by the legal system to enhance the accuracy of testimony by eyewitnesses. Apart from examining the relationship between questioning as system variable and the accuracy of testimony, the present study furthermore explores the relationship between type of questioning and certain biographical variables (occupation, age, gender and race). To achieve the aim of the study, 412 respondents consisting of 11 to 14-year-olds, university students, the public and Police College students participated and were exposed to open-ended or closed-ended questions. It was found that the participants who responded to the closed-ended questions were significantly more accurate than those who answered the open-ended questions. All the biographical groups, except the public, were more accurate in responding to the closed-ended questions. The scholars obtained the lowest scores (although not always significant) for both the open-ended and closed-ended questions. With respect to age the 18 to 25-year-olds obtained significantly higher scores than the other groups for the closed-ended questions. Whites performed significantly better than blacks in response to the open-ended and closed-ended questions.

The effect of confidence and method of questioning on eyewitness testimony.

Very often eyewitnesses are perceived as being accurate due to the confidence in the accuracy of their own testimony. The confidence displayed by an eyewitness may possibly be increased by the method of questioning used by legal professionals and police. The present study examines the confidence-accuracy relationship and the effect the method of questioning (open-ended versus closed-ended questions) may have on the confidence of eyewitnesses. The sample of 412 respondents consisted of scholars (11 to 14-year-olds), university students, the public and Police College students. A significant relationship between memory accuracy and confidence was found for more than 70% of the questions. Closed-ended questions provided a significantly higher rate of accuracy than open-ended questions. A significantly larger proportion of respondents to the closed-ended questions were more confident about their answers than those who responded to the open-ended questions.

The influence of human N-acetyltransferase genotype on the early bactericidal activity of isoniazid.

The elimination of isoniazid is subject to the influence of the N-acetyltransferase 2 (NAT2) genotype, and individuals may be homozygotic slow, heterozygotic fast, or homozygotic fast acetylators of isoniazid. The early bactericidal activity (EBA) of an antituberculosis agent can be determined by quantitative culture of Mycobacterium tuberculosis in sputum samples obtained from patients with pulmonary tuberculosis during the first days of treatment. In these studies, the EBA of isoniazid during the first 2 days of treatment was determined for 97 patients with sputum smear-positive pulmonary tuberculosis following isoniazid doses of < or =37.5 mg, 75 mg, 150 mg, 300 mg, and 600 mg. The NAT2 genotype was determined in 70 patients, and the association between EBA and genotype was examined in this subgroup. Similarly, the relationship between EBA and isoniazid serum concentration was evaluated in 87 patients. The mean EBA of isoniazid increased with dose, but it levelled off between doses of 150 mg (mean EBA, 0.572) and 300 mg (mean EBA, 0.553). Significant differences were found in the mean EBA of isoniazid between the homozygous slow acetylator group and the heterozygous fast acetylator group and between the homozygous slow acetylator group and the homozygous fast acetylator group, but not between the heterozygous fast acetylator group and the homozygous fast acetylator group. The EBA appeared to reach a maximum at a 2-h isoniazid concentration of 2-3 microg/mL. These data confirm a significant effect of NAT2 genotype on the EBA of isoniazid over a range of doses.

Genome and MIC stability in Mycobacterium tuberculosis and indications for continuation of use of isoniazid in multidrug-resistant tuberculosis.

Mycobacterium tuberculosis strains resistant to two or more of the first line antituberculosis drugs (MDR) are a serious threat to successful tuberculosis control programmes. For this retrospective study, 85 follow-up drug resistant isolates from 23 patients residing in a community with a high incidence of tuberculosis were collected and the level of in-vitro resistance to antibiotics determined quantitatively. PCR-SSCP and sequencing techniques were used to screen for gene mutations associated with resistance in 31 follow-up samples from a smaller group of eight patients. DNA fingerprint analysis was done on sequential isolates to confirm identity. Although treatment had a profound effect on changes in drug resistance patterns, the MIC for a particular agent remained constant in follow-up isolates. DNA fingerprinting and mutational analysis (14 different loci) showed that the genome of MDR strains of M. tuberculosis is relatively stable during the course of therapy. The rpoB gene was the most frequently mutated structural gene involved in drug resistance and a novel C to T mutation upstream of open reading frame (ORF)1 of the inhA operon was detected. No evidence was found of the presence of strain W (New York) in this group of MDR strains. The results stress the importance of confirming individuality of strains for the accurate calculation of frequencies of particular mutations associated with drug resistance, particularly in a high incidence area. Approximately one-half (47.8%) of the patients had isolates resistant to concentrations just above the critical concentration for isoniazid (MICs of 0.2-5 mg/L). Therefore, these patients and their contacts who develop primary drug-resistant tuberculosis may respond to higher dosages of treatment which could have a considerable impact on the cost and the ease of management of resistant tuberculosis.

The early bactericidal activity of amikacin in pulmonary tuberculosis.

SETTING: Stellenbosch University, a tertiary care hospital in Cape Town, South Africa. OBJECTIVE: To determine the early bactericidal activity (EBA) of amikacin in dosages of 5 mg/kg, 10 mg/kg and 15 mg/kg body weight in comparison to that of isoniazid 6 mg/kg body weight or no drug. DESIGN: An open, randomised trial. PATIENTS: Patients with previously untreated, sputum smear-positive pulmonary tuberculosis. INTERVENTION: Patients received amikacin 5 mg/kg (12 patients), 10 mg/kg (13 patients) or 15 mg/kg (15 patients), isoniazid 6 mg/kg (9 patients) or no drug (10 patients). RESULTS: The rate of decrease in log viable colony forming units of Mycobacterium tuberculosis per ml of sputum per day during the first 2 days of treatment with amikacin 5 mg/kg, 10 mg/kg and 15 mg/kg was 0.041 (SD 0.100), 0.045 (SD 0.144) and 0.052 (SD 0.096), respectively, 0.515 (SD 0.173) in the patients receiving isoniazid 6 mg/kg, and 0.041 (SD 0.113) in those receiving no drug. The EBA found in patients receiving amikacin did not differ significantly from that of the no drug group. However, as the EBA in the no drug group was the highest ever encountered at Stellenbosch University, the mean in patients receiving drug was tested against 0 and found to differ significantly (P = 0.03), suggesting minimal activity. Mean amikacin serum concentrations 1 hour after intramuscular drug administration were 13.5 microg/ml, 26.7 microg/ml and 39.2 microg/ml in the patients receiving 5 mg, 10 mg and 15 mg per kg body weight, respectively. CONCLUSION: Despite serum concentrations well in excess of the minimal inhibitory concentration of 2-4 microg/ml, the EBA of amikacin in patients with smear-positive pulmonary tuberculosis was only just detectable.

The early bactericidal activity of streptomycin.

SETTING: Patients with sputum smear-positive, newly diagnosed pulmonary tuberculosis studied at Tygerburg Hospital, Cape Town, for their early response to streptomycin (SM). OBJECTIVE: To determine the standard early bactericidal activity (EBA), namely the fall in viable counts of tubercle bacilli in 16-hour sputum collections during the first 2 days of treatment with SM. DESIGN: Patients were randomised to logarithmically spaced daily doses of 7.5, 15 or 30 mg/kg SM. A comparison by standard biological assay methods was then made with previous estimations of the EBA of paromomycin in doses of 7.5 and 15 mg/kg. RESULTS: An EBA of 0.133 obtained with 30 mg/kg SM differed significantly from zero (P = 0.0009), while the EBAs of 0.043 with 15 mg/kg and -0.025 with 7.5 mg/kg did not so differ. A linear regression equation of EBA = -0.2587 + 0.2627 log10 dose was obtained with significant slope (P = 0.007). Paromomycin was estimated to be 1.745 more potent than SM with wide 95% confidence limits (0.6-28.6), indicating that it cannot be considered more potent than SM. CONCLUSIONS: The low EBAs show that SM has low, dose-related, bactericidal activity in cavities, consistent with results from clinical trials. If streptomycin-resistant bacilli are present, paromomycin is probably the aminoglycoside of choice.

Culture of human granulosa cells from an in vitro fertilization program: effects of extracellular matrix on morphology and cyclic adenosine 3',5' monophosphate production.

Human GCs obtained from patients undergoing an IVF procedure and treated with CC, hMG, and hCG, lack cAMP responsiveness to hCG stimulation in vitro. After 48 hours in culture, recovery of cAMP production was significantly higher in hCG-stimulated cells grown on dishes coated with ECM than in cells grown on uncoated dishes. This difference correlated with a higher degree of morphologic differentiation of cells cultured on ECM. It is concluded that ECM provides superior culture conditions for the recovery of hormone-stimulated cAMP production and the maintenance of morphologic differentiation of preovulatory GCs from stimulated cycles.

Early bactericidal activity of antituberculosis agents.

The early bactericidal activity (EBA) of an antituberculosis agent is arbitrarily defined as the fall in log(10) colony forming units (cfu) of Mycobacterium tuberculosis per ml sputum per day during the first 2 days of treatment. Determining the EBA is an important preliminary step in the clinical evaluation of an antituberculosis agent. We review the results of eight published studies of the EBA of different antituberculosis agents, the impact of these results on our understanding of the actions of the respective agents, the clinical characteristics and sputum findings of patients included in these studies, and explore sources of variation in the EBA results. Patients in these studies had a mean age of 31-36 years, a mean weight of 50-57 kg, 67% were male and 56% had lung involvement covering an area of more than one lung, and 90% had multicavitary disease. None of these findings were related to EBA in any study. The mean log(10) cfu per ml sputum in the first specimen was 6.474. This was related to radiological extent of disease and cavity size in one study (p < 0.001) and, in the case of isoniazid to EBA with a rise in EBA of 0.094 (95% CL 0.029-0.158) for each tenfold rise in cfu counts/ml sputum. The overall variation in EBA in these studies was 0.0303, that due to laboratory processing of specimens was 0.0011, and due to patient characteristics and sputum sampling 0.0212. The EBA is a reproducible investigation that has contributed significantly to our knowledge of the actions and characteristics of both established and new antituberculosis agents. The greatest source of variation in EBA results appears to be that due to interpatient variation in disease characteristics and sputum sampling.

Radionuclide transfer to fruits: A critical review. Foreword.

After completion of the (Validation of Environmental Model Predictions) (VAMP) and (BIOspheric Model Validation Study) (BIOMOVS II) Programmes, the Division of Radiation and Waste Safety of the IAEA decided to promote a new co-ordinated research programme to continue activities in the area of biosphere modelling. As a result, the BIOMASS Programme on BIOsphere Modelling and ASSessment was launched in Vienna in October 1996. The Programme is scheduled to finish in October 2000. The overall objective of BIOMASS is to provide an international focal point in the area of biosphere assessment modelling. There are 3 Themes and 11 working groups in BIOMASS. The objective of the Theme 3 Fruits Working Group is to improve understanding of the uptake and transfer of radionuclides from different sources to fruit. As part of the work programme, a review was undertaken of the experimental, field and modelling information on the transfer of radionuclides to fruit.

The relationship between memory and the recall of specific details.

When eyewitnesses are exposed to an event, certain details tend to capture their attention more than other details, which ensures the better recall of the details that were observed. As little research has focussed on the relationship between certain biographical variables (occupation, age, gender, race and retention period) and specific event details, the present study attempts to shed some light on this area. A sample of 295 participants was obtained from 12- to 14-year-olds, university students, the public and Police College students that were divided into a short-term and long-term memory group. The details that were focussed on in the questionnaire were divided into obvious physiognomic, less pronounced physiognomic, clothing, circumstantial, and action-related details. Between the short-term and long-term memory groups it was found that significant differences were indicated for only certain items related to obvious physiognomic details, clothing details, circumstantial evidence and action-related details. For all the different biographical groups significant differences were found for certain items belonging to the different groups of details.

Memory accuracy of a real-life simulated incident.

Due to an awareness that both laboratory and real-life research present limitations, the present study attempts to combine the two research approaches, making use of the strengths of both methodologies. A real-life situation was created in a classroom and combined with laboratory conditions to determine the influence of certain variables (occupation, age, gender, race and retention interval) on the accuracy of eyewitness reporting. A total of 295 participants consisting of 12- to 14-year-olds, University students, the public and Police College students were divided into a short- and long-term memory group. The short-term group were assessed immediately after witnessing the incident, while the long-term group were only assessed five to six weeks later. It was found that the effect of passage of time was insubstantial; however, the performance of the short-term group had already been disconcertingly poor. The school children did not perform significantly less well than other occupational groups, and the members of the Police did not perform significantly better than the other groups. No significant differences were found for the different gender and racial groups.