A new malaria vector in Africa: Predicting the expansion range of Anopheles stephensi and identifying the urban populations at risk.

In 2012, an unusual outbreak of urban malaria was reported from Djibouti City in the Horn of Africa and increasingly severe outbreaks have been reported annually ever since. Subsequent investigations discovered the presence of an Asian mosquito species; Anopheles stephensi, a species known to thrive in urban environments. Since that first report, An. stephensi has been identified in Ethiopia and Sudan, and this worrying development has prompted the World Health Organization (WHO) to publish a vector alert calling for active mosquito surveillance in the region. Using an up-to-date database of published locational records for An. stephensi across its full range (Asia, Arabian Peninsula, Horn of Africa) and a set of spatial models that identify the environmental conditions that characterize a species' preferred habitat, we provide evidence-based maps predicting the possible locations across Africa where An. stephensi could establish if allowed to spread unchecked. Unsurprisingly, due to this species' close association with man-made habitats, our maps predict a high probability of presence within many urban cities across Africa where our estimates suggest that over 126 million people reside. Our results strongly support the WHO's call for surveillance and targeted vector control and provide a basis for the prioritization of surveillance.

The Huntingdon Day Hospital Trial: secondary outcome measures.

OBJECTIVE: To compare day hospital to day centre rehabilitation using scales to measure mobility, activities of daily living and quality of life. DESIGN: Single blind randomized controlled trial with home assessments at baseline (twice), six weeks and three months. SETTING: Mainly rural health district. Day hospital and social services day centres in market towns. INTERVENTIONS: Day hospital treatment or day centre rehabilitation by a physiotherapist and two health support workers. MAIN OUTCOME MEASURES: World Health Organization mobility scale scored with and without aid, Nottingham Extended Activities of Daily Living Scale and Nottingham Health Profile. SUBJECTS: One hundred and five physically disabled older patients living at home referred for day hospital rehabilitation or maintenance before discharge from hospital (66) or referred as outpatients (39). RESULTS: At three months there were no statistically significant differences between rehabilitation at day hospital and day centre for any of the outcome measurements. However, there were significant improvements between baseline and three months for the following subscales [mean change per six-week period (95% confidence interval) ]: WHO mobility subscale (with aid) -0.67 (-0.99,-0.35); Nottingham Health Profile mobility subscale -10 (-15.5,-4.5) Nottingham extended ADL mobility subscale +3.08 (1.78,4.37); Nottingham extended ADL leisure subscale +1.66 (0.96,2.36). CONCLUSION: There were no differences between day hospital and day centre in the outcomes measured. Day rehabilitation appeared to improve functional ability and mobility and scales reflecting these domains deserve further evaluation as outcome measures in this patient group. However, no improvement in quality of life was observed.

A randomized controlled trial of day hospital and day centre therapy.

OBJECTIVE: To compare the outcome of day hospital to day centre rehabilitation. DESIGN: Single blind randomized controlled trial with home assessments at baseline (twice), six weeks and three months. SETTING: Mainly rural health district. Day hospital and social services day centres in market towns. SUBJECTS: One hundred and five physically disabled older patients living at home referred for day hospital rehabilitation or maintenance before discharge from hospital (66) or referred as outpatients (39). INTERVENTIONS: Day hospital treatment or day centre rehabilitation by a physiotherapist and two health support workers. MAIN OUTCOME MEASURES: Barthel Index, Philadelphia Geriatric Morale Scale and Caregiver Strain Index. RESULTS: More day centre (23/55) than day hospital patients (6/50) (p <0.001) withdrew from allocated treatment by choice or because of operational difficulties. Both groups improved significantly in functional ability and reduction of care-giver strain by three months but there was no significant difference between groups. The mean improvement in Barthel Index (standard error) for day hospital = +1.5 (0.41) (n = 34) and day centres = +1.5 (0.48) (n = 38). The mean difference (95% confidence interval) between day hospital and day centre was 0 (-1.28, +1.28). Likewise the mean Philadelphia Geriatric Morale Scale improvement for day hospital +1.8 (0.66) (n = 35) and day centres was +0.9 (0.63) (n = 38). The mean difference was -0.88 (-2.7, +0.95). The mean reduction in Caregiver Strain for day hospital was -1.45 (0.5) (n = 23) and day centre was -1.59 (0.47) (n = 27). The difference was -0.14 (1.52, +1.24). (These analyses are all on an intention-to-treat basis.) CONCLUSION: Whilst the improvement in functional ability and care-giver strain was similar in both groups, day centre rehabilitation was less popular and had practical difficulties. If these difficulties can be overcome the model should be tested elsewhere.

Determination of inorganic anions in water by ion chromatography: a collaborative study.

The U.S. Environmental Protection Agency (U.S. EPA) and the American Society for Testing and Materials (ASTM) conducted a joint collaborative study validating an ion chromatographic method for determination fo inorganic anions (U.S. EPA method 300.0A and the equivalent proposed revision to ASTM method D4327). This study was conducted to determine the mean recovery and precision of analyses for bromide, chloride, fluoride, nitrate, nitrite, orthophosphate, and sulfate in reagent water, drinking water, and wastewater. The study design was based on Youden's nonreplicate plan for collaborative tests of analytical methods. The test waters were spiked with the anions at 6 concentration levels, prepared as 3 Youden pairs. The 22 volunteer laboratories were instructed to dilute 10 mL sample concentrate to 100 mL test water. A measured volume of sample (20-200 microL) was injected into an ion chromatograph equipped with a guard column, anion exchange column, and a chemical micromembrane suppression device. The anions were then separated using 1.7 mM sodium bicarbonate and 1.8 mM sodium carbonate, and measured by a conductivity detector. Submitted data were evaluated using U.S. EPA's IMVS computer program, which follows ASTM D2777-86 statistical guidance. U.S. EPA method 300.0A and ASTM method D4327 were judged acceptable for measurement of the above anions (except sulfate) at concentrations ranging from 0.3 to 25 mg/L and sulfate concentrations from 2.9 to 95 mg/L. Mean recoveries for the 7 anions from all matrixes, as estimated from the linear regression equations, ranged from 95 to 104%. At concentrations above 2-6 mg/L for bromide, fluoride, nitrate, nitrite, and orthophosphate, and above 24 mg/L for sulfate, the overall and single-analyst relative standard deviations were less than 10 and 6%, respectively. As concentrations decreased, precision became more variable. The relative standard deviations of results for chloride were slightly higher than the other anions, especially in matrixes with high chloride background. Analysis of Variance (ANOVA) tests at the 95% confidence interval indicated a statistically significant matrix effect for chloride, nitrite, and nitrate analyses in drinking water compared to analyses in reagent water. Because these matrix effects were caused by the spiking process and not the drinking water itself, the ANOVA determination was not considered to be of practical significance.

Optimized preparation of Aspergillus fumigatus extracts for allergy diagnosis.

Extracts of Aspergillus fumigatus are required for the measurement of specific antibodies that are important indices in the diagnosis of allergic bronchopulmonary aspergillosis (ABPA). This study investigated the effect of different culture conditions on the production and release of antigenic and allergenic proteins of A. fumigatus. Increasing the incubation temperature from 25 degrees C to 37 degrees C altered the production of proteins by the mycelium which resulted in the release of a greater number of proteins that reacted with precipitating antibodies. Static sporulating cultures produced a much wider antigenic spectrum than shake cultures although the number of precipitating proteins (5 and 3 respectively) and major IgE binding proteins (5 and 3 respectively) was not greatly altered. The widest range of proteins bound by precipitating antibody or IgE from ABPA serum were released into the culture filtrate during 28 day static incubation at 37 degrees C. The resultant extract proved useful for screening patients for specific IgE and will provide a starting material for the identification of individual antigens or allergens.

Gas chromatographic/electron capture detection method for determination of chlorinated acids in water: collaborative study.

A U.S. Environmental Protection Agency (USEPA) interlaboratory method validation study was conducted on USEPA Method 515.1, "Determination of Chlorinated Acids in Water by Gas Chromatography with an Electron Capture Detector." This method is one of the 6 pesticide methods developed for the USEPA National Pesticide Survey (NPS). Method recovery and precision for analyses of sub-ppb to low-ppb concentrations of chlorinated acids were determined in reagent water and finished drinking waters. The analytes evaluated in the study included the 12 pesticides that were quantitatively measured in the National Pesticide Survey (bentazon, 2,4-D, 2,4-DB, 3,5-dichlorobenzoic acid, DCPA-diacid, dicamba, dichlorprop. 5-hydroxydicamba, pentachlorophenol, picloram, 2,4,5-T, and 2,4,5-TP) and 5 pesticides (acifluorfen, chloramben, dalapon, dinoseb, and 4-nitrophenol) that were only qualitatively assessed in the National Pesticide Survey because of recognized method imprecision. The study design was based on Youden's nonreplicate plan for collaborative tests of analytical methods. The waters were spiked with 17 chlorinated acids, each at 6 concentration levels, prepared as 3 Youden pairs. Eight laboratories extracted the spiked test waters at pH < 2 with ethyl ether, performed a solvent exchange with methyl tert-butyl ether, prepared methyl esters of the extracted acids using diazomethane, and analyzed an aliquot of each derivatized extract by gas chromatography with electron capture detection. The submitted data were analyzed using a USEPA computer program, which measured recovery and precision for each of the 17 compounds and compared the performance of the method between water types. Method 515.1 was judged acceptable for the 12 NPS analytes recovered quantitatively; mean percent recoveries at 10-15 times the method detection limits ranged from 79 to 105% in reagent water and from 75 to 123% in finished drinking water. In reagent water, overall precision (reproducibility relative standard deviation, RSDR) ranged from 9.6 to 34.2% and in finished drinking water, the RSDR ranged from 11.9 to 37.0%. Single-analyst precision (RSD for repeatability, RSDr) ranged from 5.8 to 17.7% in reagent water and from 4.6 to 27.9% in drinking water. Results for the 5 other NPS analytes were too inaccurate or imprecise and, for these compounds, supported use of the method for qualitative measurements only; the 5 compounds are not included in the adopted method. The method has been adopted first action by AOAC INTERNATIONAL for determination of residues of 12 chlorinated acids in finished drinking water.

Gas chromatographic/nitrogen-phosphorus detection method for determination of ethylene thiourea in finished drinking waters: collaborative study.

A joint U.S. Environmental Protection Agency (USEPA)-AOAC interlaboratory method validation study was conducted on USEPA National Pesticide Survey (NPS) Method 6, "Determination of Ethylene Thiourea (ETU) in Finished Drinking Water by Gas Chromatography with a Nitrogen-Phosphorus Detector." The purpose of the study was to determine and compare the mean recoveries and precision for determination of ETU in reagent water and finished drinking waters. The study design was based on Youden's nonreplicate plan for collaborative tests of analytical methods. The waters were spiked with ETU at 6 concentrations levels, prepared as 3 Youden pairs. In the method, the test water is extracted by passing the sample through an absorbent matrix type tube. ETU is recovered from the tube with methylene chloride, the extract is solvent-exchanged to ethyl acetate, and an aliquot of each extract is analyzed by gas chromatography using a nitrogen-phosphorus detector. Twelve laboratories participated in the study. Data were analyzed using a USEPA computer program, which measured recovery and precision for ETU and compared the performance of the method between the 2 water types. Over the concentration range tested, the mean percent recoveries of ETU were 82-92% in reagent water and 85-98% in finished drinking water. The range of the between-laboratory relative standard deviations (RSDR) for the 6 concentrations was 5-24% in reagent water, but was only 4-9% in finished drinking water. The range of the within-laboratory relative standard deviations (RSDr) was 6-14% for reagent water and 6-10% for finished drinking water. Results for the 2 water matrixes showed no statistically significant differences.(ABSTRACT TRUNCATED AT 250 WORDS)

Aspergillus fumigatus: identification of 16, 18, and 45 kd antigens recognized by human IgG and IgE antibodies and murine monoclonal antibodies.

The immunochemical properties of antigens produced by Aspergillus fumigatus were investigated with biochemical purification techniques in conjunction with the production of murine monoclonal antibodies (MAbs) and binding studies with human IgG and IgE antibodies. A. fumigatus antigens were partially purified by gel filtration and hydrophobic interaction chromatography on phenyl-Sepharose. Two fractions that eluted with either 2 mol/L or 0.15 mol/L of NaCl demonstrated strong binding to human IgG and IgE antibodies. Immunoprecipitation analysis with IgG antibodies from six patients with different Aspergillus-related diseases demonstrated that the 2M and 0.15M fractions contained major antigens of molecular weight 18 kd (Asp f I) and 45 kd, respectively. The 125I-labeled 2M fraction was used to compare IgG antibodies to A. fumigatus in sera from 25 patients with Aspergillus-related diseases. IgG antibodies were significantly higher in patients with allergic bronchopulmonary aspergillosis (geometric mean, 437 U/ml) than in patients with asthma (geometric mean, 14 U/ml; p less than 0.001), but undetectable (less than 5 U/ml) in 43/48 control subjects. A good correlation was found between levels of IgG antibodies to the 125I-labeled 0.15M fraction and the 125I-labeled 2M fraction in sera from 106 patients with cystic fibrosis (r = 0.77; p less than 0.001). Five murine IgG MAbs and two IgM MAbs were raised against the 2M fraction, and immunoprecipitation with the IgG MAb demonstrated two distinct antigens within the 2M fraction, Asp f I, and a 16 kd antigen. The results of a solid-phase RIA with IgG MAb 4A6 demonstrated that approximately 85% of A. fumigatus-allergic patients with allergic bonchopulmonary aspergillosis had IgE antibodies to Asp f I. The three protein antigens defined in these studies are useful probes for investigating the immunopathogenesis of diseases associated with colonization by A. fumigatus.

Allergy to rabbits. III. Further identification and characterisation of rabbit allergens.

Crossed immunoelectrophoresis and crossed radio-immunoelectrophoresis techniques have been employed for analysis of extracts of various rabbit source materials to identify rabbit allergens in addition to the already described Antigen R1 (AgR1). Urine and dander extracts were found to contain only low levels of AgR1 and its presence in urine was as a contaminant due to mode of collection--it was not present in urine collected directly from the bladder. Other allergens were only recognised by highly rabbit-sensitive individuals, one in particular (Ag2) being present in several source materials. Serum albumin proved to be of minor allergenic importance and except for dander its presence was only in minimal amounts. As both AgR1 and Ag2 are significant components in extracts of fur and dust these extracts are therefore most recommended for use in investigations of individuals sensitive to rabbits.

Capillary column gas chromatography with nitrogen-phosphorus detection for determination of nitrogen- and phosphorus-containing pesticides in finished drinking waters: collaborative study.

A joint U.S. Environmental Protection Agency/AOAC interlaboratory method validation study was conducted on EPA Method 507, Determination of Nitrogen- and Phosphorus-Containing Pesticides in Finished Drinking Water by Gas Chromatography with a Nitrogen-Phosphorus Detector, to determine the mean recovery and precision for analyses of 45 nitrogen- or phosphorus-containing pesticides in reagent water and finished drinking waters. The study design was based on Youden's nonreplicate plan for collaborative tests of analytical methods. The waters were spiked with 45 nitrogen- or phosphorus-containing pesticides at 6 concentration levels, prepared as 3 Youden pairs. Ten volunteer laboratories extracted the spiked test waters with methylene chloride, performed a solvent exchange with methyl tert-butyl ether, and analyzed an aliquot of each extract by gas chromatography using a nitrogen-phosphorus detector. Results were analyzed using an EPA computer program, which measured recovery and precision for each of the 45 pesticides and compared the performance of the method between water types. Method 507 was judged acceptable for all analytes tested except merphos, which thermally decomposed in the injection port of the gas chromatograph. Five compounds (carboxin, disulfoton, metolachlor, pronamide, and simazine) exhibited statistically significant matrix effects for the finished drinking water. The method has been adopted official first action by AOAC.

Direct aqueous injection-liquid chromatography with post-column derivatization for determination of N-methylcarbamoyloximes and N-methylcarbamates in finished drinking water: collaborative study.

An interlaboratory method validation study was conducted on EPA Method 531.1, Measurement of N-Methylcarbamoyloximes and N-Methylcarbamates in Water by Direct Aqueous Injection HPLC with Post Column Derivatization, to determine the precision and mean recovery for determination of 10 carbamate pesticide compounds in reagent water and in finished drinking waters. The study design was based on Youden's nonreplicate plan for collaborative tests of analytical methods. The waters were spiked with 10 carbamate pesticides at 6 concentration levels, as 3 Youden pairs. Eight laboratories analyzed the samples by direct aqueous injection, with separation by reverse-phase liquid chromatography and post-column hydrolysis of the carbamates and carbamoyloximes to methylamine, followed by reaction of the methylamine with o-phthalaldehyde and 2-mercaptoethanol using fluorescence detection. Results were analyzed using an EPA computer program, which measured precision and recovery for each of the 10 compounds and compared the performance of the method between water types. The method was acceptable for all analytes tested. After removal of a nonrepresentative data set for aldicarb sulfoxide, no matrix effects were observed; the statistics for the pooled drinking waters were not significantly different from the statistics for the reagent waters. The method has been adopted official first action by AOAC.

Measurement of airborne mite antigen in homes of asthmatic children.

The airborne concentration of major house dust mite antigen Der p1 was measured by low volume sampling (2 litres/min) in the homes of 68 allergic, asthmatic children. The presence of detectable airborne antigen was strongly associated with sensitivity to the mite, whereas there was no significant relation between sensitivity and the previously recommended threshold level of 2 micrograms Der p1 per g carpet dust. There was a significant association with lower threshold levels in carpet dust (0.5 microgram/g) but at no level was the association as strong as that with air measurements. Concentrations of airborne antigen were higher in rooms with wool carpets than in those with synthetic carpets or hard floors, but there was no significant difference between the dust levels of Der p1 in the two carpet types. Air sampling is a more appropriate method of assessing antigen exposure than dust sampling for asthmatic patients.

Controlled trial of natamycin in the treatment of allergic bronchopulmonary aspergillosis.

Allergic bronchopulmonary aspergillosis often requires treatment with oral corticosteroids to control the host response to Aspergillus fumigatus. In a double blind study 25 patients with allergic bronchopulmonary aspergillosis taking maintenance oral corticosteroids were randomly allocated to receive 5 mg natamycin or placebo by nebuliser twice daily for one year. The primary aim of the study was to assess the steroid sparing potential of natamycin. Standardised reductions in corticosteroid dosage were therefore undertaken every five weeks, unless clinically contraindicated. Five patients were withdrawn in the first four months: two (1 natamycin, 1 placebo) died, two (1 natamycin, 1 placebo) had suspected drug reactions, and one (natamycin) was non-compliant. The pretreatment characteristics of the 20 patients (10 in each group) who completed the study were similar, 17 (9 natamycin, 8 placebo) having evidence of recent disease activity. At the end of the study prednisolone dose had been reduced by a similar amount in each group (median natamycin 2.25 mg, placebo 2.5 mg). Evidence of disease activity during the study year (transient shadowing on the chest radiograph, blood eosinophilia, or increases in antibodies to A fumigatus, or any combination of these) was observed in similar numbers of patients in each group (5 natamycin, 7 placebo). There was no evidence that natamycin conferred benefit on these patients with allergic bronchopulmonary aspergillosis.

Allergy to mice. II. Further characterization of two major mouse allergens (AG 1 and AG 3) and immunohistochemical investigations of their sources.

Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and quantitative immunoelectrophoretic techniques have been used to characterize further the two major mouse allergens, antigen 1 (Ag 1) and antigen 3 (Ag 3). Gel filtration using Sephacryl S-200 showed Ag 1 to have a molecular weight of 18 kD and Ag 3 of 21 kD. SDS-PAGE followed by Western blotting onto nitrocellulose then incubation with individual antisera directed against each of the two major allergens, and an alkaline phosphatase enzyme system, was used to distinguish between the two allergens and indicated a molecular weight of 17 kD for Ag 1 and 16 kD for Ag 3. Ag 3 but not Ag 1 was shown to contain polysaccharide residues. Immunohistochemical staining of mouse skin sections demonstrated that antigens detected in whole dust extracts were present in the hair follicles, on the hair shafts and on the stratum corneum. Staining of similar sections using the rabbit anti-Ag 3 showed the presence of this major allergen in the hair follicles coating the hairs and extending along the skin surface. Serum from a pool of mouse-allergic subjects also demonstrated staining in the same areas when detected using a fluorescein-labelled anti-human IgE as second antibody. As both major allergens were present in extracts of fur this would appear to be most appropriate for use in diagnosis (i.e. skin test and RAST) and also possibly desensitization. However, dust from isolators (available in greater amounts) would be equally suitable.

Pigeon breeders' disease: quantitative immunoelectrophoretic studies of pigeon bloom antigen.

This study employed quantitative immunoelectrophoretic techniques, on sera from confirmed cases of pigeon breeders' disease (PBD), to investigate the antigenicity of a pigeon bloom extract, implicated as a sensitizing agent in this disease. On crossed immunoelectrophoresis the maximum number of antigenic components identified was 29 for the bloom compared to 10 for pigeon serum. A major component was shown to be closely related to pigeon IgA, and demonstrated partial crossreactivity to the pigeon IgG. This component also showed identity with the major component of a pigeon droppings extract, considered to be derived from intestinal IgA. Only trace amounts of serum albumin were detected and most other bloom components were not serum-related. Although greater overall antigenic similarity was found to pigeon droppings extract, at least three of the bloom components appeared to be specific. The bloom extract also contained a low amount of an alpha-techoic acid-like component, causing some non-specific reactivity. Pigeon feather dust or 'bloom', like pigeon droppings, is therefore a potent source of antigens associated with PDB--pigeon IgA being a major component of both antigens.

Complex microphthalmos.

Forty patients were diagnosed as having complex microphthalmos on the basis of a malformed globe with a total axial length measurement at least 2 SDs below the mean for age-similar controls. Three had anterior segment dysgenesis; 4, congenital lens abnormalities; 14, chorioretinal colobomas; 12, persistent hyperplastic primary vitreous; 4, retinal dysplasia; and 3, complex malformations due to ipsilateral facial malformations. Measurements of total axial length indicated that complex microphthalmos was congenital and that postnatal growth of the malformed eye was similar to that of normal eyes. In most patients the anterior segment length was normal, while in all patients the posterior segment length was at least 2 SDs below the mean. Corneal diameter correlated significantly with total axial length (r2 = .57) and decreased linearly as total axial length decreased. In most patients in whom measurements were obtained, the lens and corneal power were increased, thereby compensating for decreased total axial length. We propose that inadequate production of secondary vitreous is the cause of the microphthalmos, given that the posterior segment was disproportionately reduced in size and the secondary vitreous is its predominant component. Evidence that each of the various ocular malformations can influence the production of secondary vitreous is presented.

Simple microphthalmos.

Simple microphthalmos was diagnosed in 22 patients on the basis of a normal-appearing eye and a total axial length at least 2 SDs below the mean for age. Anterior segment length was normal in most patients while posterior segment length was at least 2 SDs below the mean in all patients, indicating that disproportionate reduction in posterior segment length accounted for the microphthalmos. The normal values for total axial length, anterior segment length, and posterior segment length were determined from the analysis of axial length measurements obtained from age-similar controls. Ten patients had isolated microphthalmos. One of them was diagnosed as having nanophthalmos on the basis of microcornea, total axial length less than 18 mm, and absence of systemic disease. Twelve patients had associated systemic disorders, such as fetal alcohol syndrome, myotonic dystrophy, and achondroplasia, which implicated decreased size of the optic cup, altered vitreous proteoglycans, low intraocular pressure, and abnormal release of growth factors in the pathogenesis of microphthalmos.

IgG antibodies in relation to exposure to laboratory-animal allergens.

An ELISA technique has been used to measure levels of specific IgG in individuals exposed to laboratory animals in comparison to levels in unexposed individuals. The technique was to determine whether these levels could be useful as a monitor of exposure. Both symptomatic (with and without specific IgE) and nonsymptomatic individuals were studied. Several antigenic source materials were investigated for each animal. The IgG response was extremely complex and elicited a varied pattern of response between animals (mouse, rat, and rabbit). No evidence was found for raised IgG levels being directly related to the degree of exposure, more to the presence of clinical symptoms and specific IgE. Long-term heavy exposure to rats in nonsymptomatic individuals appeared to produce a suppression of the IgG response in relation to control levels. These findings suggest that IgG is not an indicator of exposure but demonstrate a varied response related to the presence of specific allergy to an animal, the particle size on which antigens are carried, and the physiologic state of the mucosa onto which the particles impact.