Travelers were subsequently contacted by telephone within a week of their return to minimize recall bias. Individuals were considered lost to follow-up after three unsuccessful calls at 1-week intervals. Data regarding risk behaviors, the occurrence

of health problems during travel, and malaria chemoprophylaxis observance were recorded. Data regarding insect bite prophylaxis, sun exposure, food and drink consumption, freshwater bathing, sport activities, wet sand exposure, and animal contact were documented. The occurrence of health problems during travel was recorded. Systematically, investigation was BIBF 1120 chemical structure conducted for the following: fever, cough, nose and throat diseases, diarrhea, vomiting, dehydration, heat stress, chronic disease decompensation, lower limb venous problems, trauma, psychological disorders, genitourinary symptoms, and skin diseases, including insect bites and sunburns. Data were analyzed with the SPSS v15.0 (SPSS, Inc., Chicago, IL, USA) software package. Chi-square tests were used to compare proportions of travelers who reported specific symptoms to those who did not. A p value <0.05 was considered significant. All p values were determined by two-tailed t-test. Factors associated with poor

compliance to malarial prophylaxis were explored using logistic regression models. Factors with p values below 0.20 in univariate models were considered eligible for multivariate analysis, as suggested in the classical work of Mickey and Greenland.11 A stepwise procedure based on likelihood Tacrolimus manufacturer ratio criteria was used to obtain the best criteria with the lowest Akaike criteria.12–14 Acetophenone For the final model, a two-tailed p value <0.05 was considered significant. Data were prospectively collected from the GeoSentinel data platform, using standard GeoSentinel data fields,15 for patients presenting to the two sites in Marseille (Infectious Diseases and Tropical Medicine wards, Hôpital Nord and Hôpital Lavéran) from March 2003 to December 2008 with a travel-associated illness

following travel to Senegal. The GeoSentinel Surveillance Network consists of specialized travel/tropical medicine clinics on six continents where ill travelers are seen during or after traveling to a wide range of countries and where information on travelers is prospectively recorded using a standardized format (www.geosentinel.org). Information collected included demographic data (age, sex, and country of birth), reason for most recent travel, duration of travel, pre-travel encounter, and time to presentation. Patients whose reason for traveling was their initial migration trip from Senegal to France were excluded from the study. Among the 392 individuals enrolled during pre-travel consultation, nine canceled their journey (2.3%), 25 were lost in follow-up (6.4%), and 358 were administered a post-travel questionnaire.

This can be accomplished using solid media (e.g. agar or uncompacted soil). For soil studies, Ljungholm et al. (1979) have demonstrated that the problem can be readily solved. They closed their ampoules with a 1-mm-thick porous silicone rubber seal because the material readily transmits simple gases. This procedure was shown to allow sufficient gas exchange (O2 and CO2), without significant loss of water, between the calorimetric ampoule and the atmosphere. Similarly, addition of glucose as a powder and not as selleck inhibitor a solution to soil samples combined with

the use of a flow-through cell is also a simple means to achieve calorimetric measurements in soil samples (Sparling, 1983) without reaching oxygen depletion. Finally, it is also possible to calculate the amount of oxygen present in the headspace of the calorimetric ampoule and calculate the amount of substrate that can be consumed using this oxygen. Using such simple calculations, Vor et al. (2002) were able to estimate when the transition from oxic to anoxic conditions in soil samples occurred and study changes in the metabolic heat production associated with this transition. Similarly, the use of agar medium or other solid growth substrates allows microorganisms to grow on top of the medium and therefore remain in contact with oxygen PFT�� price present in the headspace (Wadsöet al., 2004).

Furthermore, a closed environment can also be analytically advantageous – for mass balance calculations for example. Finally, it must be noted that the heat flow signal is a nonspecific, net signal related to the sum of all chemical and physical processes taking place in an IMC Non-specific serine/threonine protein kinase ampoule. As a consequence, unknown phenomena may produce some of the heat measured, and there may be simultaneous exothermic and endothermic processes taking place (Lewis & Daniels, 2003). However, well-described phenomena can be studied

under controlled conditions with a high accuracy [see the ‘diauxie’ (Monod, 1949) example in Fig. 1, Table 2]. Careful planning of IMC experiments is of great importance. Logical experimental designs must be devised and used that ensure that the observed heat flows are directly related to the processes of interest. IMC has been used in many different fields of microbiology. Medical and environmental applications provide an indication of the possibilities. One noteworthy medical application is rapid isothermal microcalorimetric detection of bacterial infection or contamination, which is of critical importance in quickly implementing the correct treatment. Recent studies have shown that with IMC, it is possible to detect bacterial contamination of donated blood platelets within a few hours (Trampuz et al., 2007). Similarly, it is also possible to determine inhibitory effects and/or the minimal inhibitory concentration for different antimicrobial compounds and microorganisms within hours using IMC (Xi et al., 2002; Yang et al., 2008; von Ah et al., 2009).

This can be accomplished using solid media (e.g. agar or uncompacted soil). For soil studies, Ljungholm et al. (1979) have demonstrated that the problem can be readily solved. They closed their ampoules with a 1-mm-thick porous silicone rubber seal because the material readily transmits simple gases. This procedure was shown to allow sufficient gas exchange (O2 and CO2), without significant loss of water, between the calorimetric ampoule and the atmosphere. Similarly, addition of glucose as a powder and not as BYL719 chemical structure a solution to soil samples combined with

the use of a flow-through cell is also a simple means to achieve calorimetric measurements in soil samples (Sparling, 1983) without reaching oxygen depletion. Finally, it is also possible to calculate the amount of oxygen present in the headspace of the calorimetric ampoule and calculate the amount of substrate that can be consumed using this oxygen. Using such simple calculations, Vor et al. (2002) were able to estimate when the transition from oxic to anoxic conditions in soil samples occurred and study changes in the metabolic heat production associated with this transition. Similarly, the use of agar medium or other solid growth substrates allows microorganisms to grow on top of the medium and therefore remain in contact with oxygen E7080 present in the headspace (Wadsöet al., 2004).

Furthermore, a closed environment can also be analytically advantageous – for mass balance calculations for example. Finally, it must be noted that the heat flow signal is a nonspecific, net signal related to the sum of all chemical and physical processes taking place in an IMC DOCK10 ampoule. As a consequence, unknown phenomena may produce some of the heat measured, and there may be simultaneous exothermic and endothermic processes taking place (Lewis & Daniels, 2003). However, well-described phenomena can be studied

under controlled conditions with a high accuracy [see the ‘diauxie’ (Monod, 1949) example in Fig. 1, Table 2]. Careful planning of IMC experiments is of great importance. Logical experimental designs must be devised and used that ensure that the observed heat flows are directly related to the processes of interest. IMC has been used in many different fields of microbiology. Medical and environmental applications provide an indication of the possibilities. One noteworthy medical application is rapid isothermal microcalorimetric detection of bacterial infection or contamination, which is of critical importance in quickly implementing the correct treatment. Recent studies have shown that with IMC, it is possible to detect bacterial contamination of donated blood platelets within a few hours (Trampuz et al., 2007). Similarly, it is also possible to determine inhibitory effects and/or the minimal inhibitory concentration for different antimicrobial compounds and microorganisms within hours using IMC (Xi et al., 2002; Yang et al., 2008; von Ah et al., 2009).

All patients who started ART remained on ART until the end of follow-up, although two participants switched to second-line ART by the end of the follow-up period. Since 1995, all HIV-positive participants not on

ART have had a CD4 cell count measurement taken every 6 months using FACSCount (Becton Dickinson, San Jose, CA, USA). Between 1992 and 1995, CD4 cell counts were determined in an external laboratory using flow cytometry. Viral load was selleck kinase inhibitor measured once a year in HIV-positive participants not on ART using the Bayer Quantiplex HIV RNA Branched DNA 3.0 assay (Bayer Diagnostics, Emeryville, CA, USA). For participants on ART, CD4 cell counts were performed at baseline and then every 3 months. Viral load was determined at baseline and every 6 months. Cryptococcal meningitis was diagnosed using Indian ink staining and culture of cerebrospinal fluid; PCI-32765 mouse cryptosporidial diarrhoea was diagnosed using modified Ziehl–Neelsen staining

of stools; other bacterial infections were diagnosed on clinical grounds as well as by culture of relevant specimens; toxoplasmosis was diagnosed on the basis of clinical presentation and/or serum toxoplasma antibody titres; and Pneumocystis jirovecii pneumonia was diagnosed on clinical grounds. Statistical analysis was performed using stata 10.0 (StataCorp, College Station, TX, USA). Participants who attended at least once following the enrolment visit contributed person-time at risk for the analysis. Patients were censored at death, at the date of their last clinic visit if they were lost to follow-up or transferred out of the study area or at 31 December 2008. Patients who were ‘lost to follow-up’ were patients not known to have died and who last attended a clinic appointment on or before 30 April 2008; 8 months prior to the

Progesterone end of the follow-up period on 31 December 2008. We assumed that individuals were at risk only once for herpes zoster virus eruption, oral hairy leukoplakia, persistent generalized lymphadenopathy, Kaposi sarcoma, HIV encephalopathy, and weight loss >10% of body weight. For other conditions, an individual experiencing an event was deemed not to be at further risk of that event for a specified period, following which they became at risk again. We assumed that individuals were not at risk for 30 days from the onset of an episode of severe bacterial infection (including pneumonia), oesophageal candidiasis and salmonellosis; for 90 days from the onset of cerebral toxoplasmosis, extrapulmonary cryptococcus, unexplained chronic diarrhoea lasting 1 month or more and unexplained prolonged fever lasting 1 month or more; for 14 days from the onset of minor mucocutaneous conditions; for 7 days from the onset of recurrent upper respiratory tract infections and oral candidiasis; for 6 weeks from the onset of Pneumocystis jirovecii pneumonia; and for 6 months from the onset of pulmonary and extrapulmonary tuberculosis.

The tubes were then visually screened for alterations in the intensity Seliciclib of the purple-colored reaction product. Oxalate measurements were performed using the Sigma oxalate diagnostic kit (catalog no. 591-D; St. Louis, MO), according to the manufacturer’s instructions. In brief, the oxalate was oxidized by oxalate oxidase to carbon

dioxide and hydrogen peroxide. The hydrogen peroxide generated was then allowed to react with 3-methyl-2-benzothiazolinone hydrazone and 3-(dimethylamino)benzoic acid in the presence of peroxidase to yield an indamine dye that was read at 590 nm. Cells were removed by centrifugation before quantifying the oxalic acid levels in the media. Experiments were repeated three times. All assays were conducted in duplicate, the results were averaged, and the error was determined. Based on the Southern blot analysis (data not shown), DNA fragments of the appropriate size were cut from the gel, purified, and subcloned into pBluescript II KS-. The individual constructs were propagated in the E. coli strain, DH5α. Plasmid DNA was isolated using the Wizard selleckchem miniprep kit (Promega, Madison, WI) and sequenced (Molecular

Genetics Core Facility, Department of Microbiology and Molecular Genetics, UT-Houston Medical School, Houston, TX). Sequence analysis was conducted using the University of Wisconsin Genetic Computer Group software (Program Manual for the wisconsin package, version 8, Genetics Computer Group, Madison, WI). Database homology searches were conducted using blastx programs (NCBI). The obcA ORF was amplified by PCR using gene-specific primers 5′-ATGACATCGCTATACATCACGGCAG-3′ and 5′-TCAGCCCGCCGCGGTCTGGGGGTCG-3′. The PCR reaction was conducted using the PCRx enhancer kit (Invitrogen Life Technology) according to the manufacturer’s instructions. All hybridization steps were

performed on a PTC-200 thermal cycler (MJ Research, Watertown, MA) using the following parameters: 94 °C for 1 min, followed by 30 cycles of 94 °C for 30 s, 58 °C for 30 s, and 72 °C for 2 min. After completion of the 30 cycles, a 5-min extension was run at 72 °C. The amplified ORF was TA cloned using the Qiagen TA cloning kit (Qiagen Inc., Valencia, CA). The obcA ORF was then isolated by restriction digestion with EcoRI and subcloned into the corresponding below site in the pRK415 vector (Wang et al., 2006) for complementation of the Bod1 mutant. For complementation with the C1E2 fragment, a 9-kb EcoRI genomic DNA fragment was cloned into the corresponding site in the pRK415 vector and transformed into a Bod1 mutant. Deletions were made of the 9-kb C1E2 genomic DNA fragment using the available restriction sites and PCR. The C1E2 EcoRI fragment was subcloned into the EcoRI site of pBluescript II KS-. To generate C1E2S2, the C1E2 construct was digested with SacI and religated. To generate the C1E2S2C1, the C1E2S2 construct was digested with ClaI and religated.

As we initially hypothesised that switch trials would engage distributed networks of task-set reconfiguration and top-down attention to a greater extent than repeat trials, we sought to test for topographic differences among conditions that would suggest the differential engagement of a subset of cortical generators. To test for periods of topographic modulation irrespective of changes in oscillatory amplitude, we calculated the global dissimilarity (GD; Lehmann & Skrandies, 1980) between differential alpha-band activity (8–14 HZ) across the anticipatory period preceding Switch trials and Repeat trials. Differential activity is derived by subtracting cue-visual trials from cue-auditory

trials. GD is a method for assessing configuration differences between two scalp distributions, independent of their strength, as the data are normalised Ceritinib order using the global field power. The GD is calculated as the square root of the mean of selleck chemical the squared differences between the potentials measured at each of the 168 scalp electrodes. For each subject and time point, the GD indexes a single value, which varies between 0 and 2 (0 = homogeneity, 2 = inversion of topography). To create an empirical probability distribution against which the GD can be tested for statistical significance, the Monte Carlo manova was applied. This is a nonparametric bootstrapping procedure, wherein each subject’s data from each time

point are permutated such that they can ‘belong’ to either condition. For each time point, the dissimilarity was then calculated for each of 5000 such permutations (Manly, 1997). To provide a more

general description of the spatiotemporal properties of differential alpha-band activity as a function of task-set reconfiguration, we computed separate statistical cluster plots (SCPs) for trials preceding a Switch and Repeat of task. This procedure has been used effectively in post hoc analyses as a means to more fully explore complex datasets and generate pointed follow-up hypotheses (Molholm et al., 2002; Murray et al., 2002). Point-wise two-tailed t-tests between attend-visual and attend-auditory trials were calculated at each time-point for all electrodes. The results of the point-wise t-tests from 168 electrodes are displayed as an intensity plot to efficiently summarise and facilitate the identification LY294002 of the onset and general topographic distribution of differential alpha-band activity preceding a Switch and a Repeat of task. The x-, y-, and z-axes, respectively, represent time, electrode location and the t-test result (indicated by a color value) at each data point. For each scalp electrode, only the first time point at which the t-test exceeded the P-value criterion of 0.05 for at least 11 consecutive data points (> 20 ms at a 512 Hz digitisation rate) is considered significant (Guthrie & Buchwald, 1991; Foxe & Simpson, 2002).

23% (P = 0.0002). Dizziness and abnormal dreams/nightmares occurred significantly less frequently with rilpivirine vs. efavirenz (P < 0.01). In both groups, patients with prior neuropsychiatric history tended to report more neuropsychiatric AEs but rates BAY 80-6946 mouse remained lower for rilpivirine than for efavirenz. Rilpivirine was associated with fewer neurological and psychiatric AEs of interest than efavirenz over 48 weeks in treatment-naïve, HIV-1-infected adults. “
“The aim of the study was to assess the separate contributions of

smoking, diabetes and hypertension to acute coronary syndrome (ACS) in HIV-infected adults relative to uninfected adults. Two parallel case–control studies were carried out. In the first study, HIV-positive adults diagnosed with ACS between 1997 and 2009 (HIV+/ACS) were matched for age, gender and known duration of HIV infection with HIV-positive adults without ACS (HIV+/noACS), each individual in the HIV+/ACS group being matched with three individuals in the HIV+/noACS group. In the second study, each individual in the HIV+/ACS group in the first study was matched for age, gender and calendar date of ACS diagnosis with three HIV-negative individuals diagnosed with ACS between 1997 and 2009 (HIV–/ACS). Each individual in the

HIV–/ACS group was then matched for age and gender with an HIV-negative adult without ACS (HIV–/noACS). After matching, the ratio of numbers of individuals in the HIV+/ACS, HIV+/noACS, HIV–/ACS and HIV–/noACS groups was therefore 1 : 3 : 3 : 3, respectively. We performed logistic regression Chlormezanone analyses AZD0530 clinical trial to identify risk factors for ACS in each case–control study and calculated population attributable risks (PARs) for smoking, diabetes and hypertension in HIV-positive and HIV-negative individuals. There were

57 subjects in the HIV+/ACS group, 173 in the HIV+/noACS group, 168 in the HIV–/ACS group, and 171 in the HIV–/noACS group. Independent risk factors for ACS were smoking [odds ratio (OR) 4.091; 95% confidence interval (CI) 2.086–8.438; P < 0.0001] and a family history of cardiovascular disease (OR 7.676; 95% CI 1.976–32.168; P = 0.0003) in HIV-positive subjects, and smoking (OR 4.310; 95% CI 2.425–7.853; P < 0.0001), diabetes (OR 5.778; 95% CI 2.393–15.422; P = 0.0002) and hypertension (OR 6.589; 95% CI 3.554–12.700; P < 0.0001) in HIV-negative subjects. PARs for smoking, diabetes and hypertension were 54.35 and 30.58, 6.57 and 17.24, and 9.07 and 38.81% in HIV-positive and HIV-negative individuals, respectively. The contribution of smoking to ACS in HIV-positive adults was generally greater than the contributions of diabetes and hypertension, and was almost twice as high as that in HIV-negative adults. Development of effective smoking cessation strategies should be prioritized to prevent cardiovascular disease in HIV-positive adults.

The SHCS is a prospective observational cohort study, established in 1988, that continuously enrols and follows HIV-positive individuals aged ≥16 years at five university out-patient clinics, two cantonal hospitals, 14 affiliated regional hospitals, and 39 private practices collaborating with the university centres [24]. Laboratory, clinical and behavioural characteristics are collected at registration and at follow-up visits every 6 months. To study the smoking status, we selected cohort participants with at least one follow-up visit with available information on smoking after 1 April 2000, when information on

smoking behaviour was included in the cohort questionnaires. The SHCS was I BET 762 approved by local ethical review boards, and written informed consent was obtained from all participants. The single

centre intervention included training for HIV care physicians on smoking cessation counselling and in the pharmacotherapy of nicotine dependence, Apitolisib in vitro and a physicians’ checklist for semi-annual documentation of counselling. Between November 2007 and December 2009, all physicians at the HIV out-patient clinic at the University Hospital Zurich took part in half a day of training on smoking cessation. This training – conducted in a standardized way by trainers of the Swiss Lung Association – included information on identification of smokers, nicotine dependence, nicotine withdrawal-related problems, motivation stages of intended behavioural change of substance-dependent persons according to the Prochaska/Di Clemente transtheoretical model [19, 25], methods of counselling, and pharmacological support of smoking cessation. At every cohort visit during the intervention period, physicians had to complete

a short checklist to document the participants’ smoking status, their current motivation level to stop smoking, and physician’s support offered at this visit. Support for smoking cessation included Clomifene short or detailed counselling about problems associated with smoking cessation, information on medication (nicotine, bupropion and varenicline), arranging a follow-up appointment for further discussion about smoking cessation, and, if appropriate, planning a date for smoking cessation. According to the broadly accepted criterion of 6 months of nicotine abstinence for smoking cessation [26], we defined a smoking cessation event as at least one follow-up visit with smoking followed by at least two consecutive semi-annual follow-up visits without smoking.

Using SSH, we had isolated three fragments encoding the factors HrpF, HrpD4-HpaA, and HrpB8 in Xoo MAI1. Essential for bacteria–host interaction are hrp genes encoding proteins involved in the T3SS, as demonstrated for various plant pathogenic bacteria by different authors (Alfano & Collmer, 2004; He et al., 2004; Büttner & Bonas, 2006). HrpF is a putative translocon protein that is essential

for pathogenicity in plant-pathogenic bacteria (Büttner et al., 2002, 2007; Meyer et al., 2006; Büttner & He, 2009). The hrp regions also contain so-called hrp-associated (hpa) genes; the hpaA and hrpB genes are encoded selleck chemicals llc by the hrpD and hrpB operons, respectively. The gene hpaA is an important virulence factor that contributes to T3SS and Ruxolitinib datasheet effector protein translocation to host cells (Lorenz et al., 2008), whereas the translated sequence derived from hrpB8 is similar to the amino acid sequence of FliR, which has been determined to be a component in the T3SS flagellar export apparatus in Salmonella typhimurium (Fan et al., 1997). We also found DNA fragments that present similarity to genes encoding RTX (repeats in toxin) toxins (FI978128 and FI978182). In Bradyrhizobium elkanii, rtx genes are involved in rhizobitoxine biosynthesis, which inhibits ethylene biosynthesis in plants (Sugawara et al., 2007). Recently,

B. elkanii rtx gene homologs were found, forming gene clusters in Xoo genomes (Ochiai et al., 2005; Sugawara et al., 2007). In the animal pathogen Kingella kingae, disruption of these genes results in the loss of toxicity (Kehl-Fie & Geme, 2007). Of the 17 clones tested, 12 were present aminophylline in the Xoo strain MAI1 and absent from the corresponding

driver DNA (Xoo PXO86 and/or Xoc BLS256). Of the four fragments tested against several other X. oryzae strains from different geographical origins, one (FI978197) specifically hybridized to the Xoo strain MAI1 (data not shown). Three (FI978100, FI978105, and FI978167) yielded hybridization signals with all the African Xoo strains tested, but not with the Asian Xoo and Xoc strains (data not shown and Table 1). These fragments corresponded to genes of ‘unknown function’ and may represent specific Xoo MAI1 genes (i.e. FI978197) and/or specific genes of African strains. From the 134 SSH Xoo MAI1 nonredundant sequences, 20 were found in both libraries (Table 2). blast analysis showed that eight of these fragments correspond to hypothetical and/or unknown proteins. The remaining 12 fragments were distributed across seven functional categories (Table 2). Using blastn, the genome of Xoc strain BLS256 was searched for these 20 SSH Xoo MAI1 sequences, with 16 being found in the Xoc BLS256 genome. The ratio identities/sequence size, obtained after blast search, nevertheless indicated a low identity percentage, <50% for most cases (Table 2).

The calibrated selleckchem standard serum 89-SF was absorbed with 10 μg/mL CWPS. At least two internal controls were added to each plate. For serotypes 14 and 19F, a high-titre and a low-titre serum sample from

pneumococcal vaccine responders were used as internal controls, respectively. For serotype 23F, two high-titre and two low-titre serum samples from pneumococcal vaccine responders were used as internal controls. The coefficients of variation (CVs) for the high-titre controls were 19.53 and 16.55% for serotypes 14 and 19F and the CVs were 18.34 and 17.91% for serotype 23F. The CVs for the low-titre controls were 17.12 and 14.91% for serotypes 14 and 19F and the CVs were 15.39 and 14.23% for serotype 23F. Sera from persons with AIDS who had no antibodies to S. pneumoniae serotype 14, 19F, 23F or 6B capsules and <1 μg of antibody to CWPS per mL were used as negative controls. Capsular polysaccharides from S. pneumoniae serotypes 14 (catalogue number 197-X; lot #2038310), 19F (catalogue number 205-X; lot #2033178), 23F (catalogue number 217-X; lot #2048448) and 6B (catalogue number 225-X; lot #2045655) were obtained from the American

Type Culture Collection (ATCC; Manassas, VA, USA). These capsular polysaccharides were suspended in phosphate-buffered saline (PBS; pH 7.4) containing 0.02% NaN3 at a concentration of 10 μg/mL and used directly to coat wells

Staurosporine by incubation at 4 °C overnight. After washing of the antigen-coated plates, the pre-absorbed patient sera or controls were added to plates and incubated at room temperature for 2 h. After thorough washing to remove antibodies not bound to the wells, horseradish peroxidase (HRP)-conjugated goat antibody to human IgG (Zymed Laboratories Inc., San Francisco, CA, USA) at a dilution of 1:2000 was used to detect IgG, and the reaction was developed for 10 min in the dark by the addition of K-blue substrate (Neogen Rapamycin Corporation, Lexington, KY, USA), followed by the addition of 1N sulphuric acid to stop the reaction. Between incubations, all washings were performed with Tris buffered saline (TBS) buffer containing 0.1% Brij solution. Optical density was read in an ELISA reader (SpectraMAX 340; Molecular Devices, Sunnyvale, CA, USA) at a wavelength of 450 nm, with subtraction of the optical density of the appropriate blank. The antibody concentrations were calculated relative to the 89-SF control serum, using the US Centers for Disease Control and Prevention (CDC) program elisa for Windows (version 2.15) [28]. The detection limits for our assay for 14, 19F, 23F and 6B, determined using the control serum 89-SF, were 0.02, 0.015, 0.02 and 0.50 μg/mL, respectively.