Evaluating capacity at three government referral hospital emergency units in the kingdom of Eswatini using the WHO Hospital Emergency Unit Assessment Tool.

BACKGROUND: The Kingdom of Eswatini, a lower-middle income nation of 1.45 million in southern Africa, has recently identified emergency care as a key strategy to respond to the national disease burden. We aimed to evaluate the current capacity of hospital emergency care areas using the WHO Hospital Emergency Unit Assessment Tool (HEAT) at government referral hospitals in Eswatini. METHODS: We conducted a cross-sectional study of three government referral hospital emergency care areas using HEAT in May 2018. This standardised tool assists healthcare facilities to assess the emergency care delivery capacity in facilities and support in identifying gaps and targeting interventions to strengthen care delivery within emergency care areas. Senior-level emergency care area employees, including senior medical officers and nurse matrons, were interviewed using the HEAT. RESULTS: All sites provided some level of emergency care 24 h a day, 7 days a week, though most had multiple entry points for emergency care. Only one facility had a dedicated area for receiving emergencies and a dedicated resuscitation area; two had triage areas. Facilities had limited capacity to perform signal functions (life-saving procedures that require both skills and resources). Commonly reported barriers included training deficits and lack of access to supplies, medications, and equipment. Sites also lacked formal clinical management and process protocols (such as triage and clinical protocols). CONCLUSIONS: The HEAT highlighted strengths and weaknesses of emergency care delivery within hospitals in Eswatini and identified specific causes of these system and service gaps. In order to improve emergency care outcomes, multiple interventions are needed, including training opportunities, improvement in supply chains, and implementation of clinical and process protocols for emergency care areas. We hope that these findings will allow hospital administrators and planners to develop effective change management plans.

Exposure to solar ultraviolet radiation limits diet-induced weight gain, increases liver triglycerides and prevents the early signs of cardiovascular disease in mice.

BACKGROUND AND AIMS: Sunlight exposure is associated with a number of health benefits including protecting us from autoimmunity, cardiovascular disease, obesity and diabetes. Animal studies have confirmed that ultraviolet (UV)-B radiation, independently of vitamin D, can limit diet-induced obesity, metabolic syndrome and atherosclerosis. The aim of this study is to investigate whether exposure to the UV radiation contained in sunlight impacts on these disease parameters. METHODS AND RESULTS: We have trialled an intervention with solar UV in obese and atherosclerosis-prone mice. We have discovered that solar-simulated UV can significantly limit diet-induced obesity and reduce atheroma development in mice fed a diet high in sugar and fat. The optimal regime for this benefit was exposure once a week to solar UV equivalent to approximately 30 min of summer sun. Exposure to this optimal dose of solar UV also led to a significant increase in liver triglycerides which may protect the liver from damage. CONCLUSION: Our results show that the UV contained in sunlight has the potential to prevent and treat chronic disease at sites distant from irradiated skin. A major health challenge going forward will be to harness the power of the sun safely, without risking an increase in skin cancers.

Surface plasmon resonance-based immunoassay for human C-reactive protein.

A rapid and highly-sensitive surface plasmon resonance (SPR)-based immunoassay (IA) has been developed and validated for detecting human C-reactive protein (CRP), a specific biomarker for inflammatory and metabolic disorders, and infections. The 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC)-activated protein A/G (Pr A/G) was diluted in 1% (v/v) 3-aminopropyltriethoxysilane (APTES), dispensed on a KOH-treated gold (Au)-coated SPR chip, and incubated for 30 min. The Pr A/G functionalized Au SPR chip was then bound to anti-human CRP capture antibody (Ab), blocked with bovine serum albumin, and subsequently used for the detection of CRP. The highly-simplified oriented Ab immobilization strategy enabled the leach-proof binding of capture Ab in 5-fold shorter time than conventional procedures. The developed IA detected 1.2-80 ng mL(-1) of CRP with a limit of detection (LOD) and a limit of quantification (LOQ) of 1.2 ng mL(-1) and 4.6 ng mL(-1), respectively. It detected CRP spiked in diluted human whole blood, serum and plasma as well as the CRP levels in the ethylenediaminetetraacetic acid (EDTA) plasma samples of patients with the same precision as the clinically-accredited analyzer-based IA and conventional CRP sandwich ELISA. The Ab-bound SPR chips stored at 4 °C retained their functional activity for 10 weeks, resulting in significant reduction in the overall analysis time.

Surface plasmon resonance-based immunoassay for human fetuin A.

This article describes a highly-sensitive surface plasmon resonance (SPR)-based immunoassay (IA) for human fetuin A (HFA), a specific biomarker for atherosclerosis and hepatocellular carcinoma. The assay is based on a novel immobilization procedure that simply involves the dilution of an anti-HFA capture antibody (Ab) in 1% (v/v) 3-aminopropyltriethoxysilane (APTES), followed by its dispensing on a KOH-treated gold (Au)-coated SPR chip and incubation for 30 min. The developed SPR IA detected 0.3-20 ng mL(-1) of HFA with a limit of detection and sensitivity of 0.7 ng mL(-1) and 1 ng mL(-1), respectively. The highly-simplified Ab immobilization procedure is also 5-fold more rapid than conventional procedures. It leads to the leach-proof binding of the capture Ab, which means that the developed SPR IA is highly cost-effective, as the Ab-bound SPR chip could be reused for many repeated HFA IAs after regeneration with 10 mM glycine-HCl, pH 2.0. The Ab-bound SPR chip, stored at 4 °C, lost only 18% of its original activity after 4 months. For the detection of HFA spiked in diluted human whole blood and plasma, the results obtained by the developed SPR IA agreed well with the commercial HFA sandwich ELISA.

Temperature-programmable resistively heated micromachined gas chromatography and differential mobility spectrometry detection for the determination of non-sulfur odorants in natural gas.

A portable, fast gas chromatographic method for the direct measurement of the parts per billion level of sulfur-free odorants in commercially available natural gas is introduced. The approach incorporates a resistively heated, temperature-programmable silicon micromachined gas chromatograph that employs a standard capillary column for the fast separation of methyl and ethyl acrylate from the natural gas matrix. The separation approach is coupled to a micromachined differential mobility detector to enhance analyte detectability, and the overall selectivity obtained against the matrix is described. A complete analysis can be conducted in less than 70 s. Furthermore, these two compounds can be measured accurately in the presence of other common volatile sulfur-based odorants such as alkyl mercaptans and alkyl sulfides. Repeatability of less than 3% RSD (n = 20) over a range from 0.5 to 5 ppm was obtained with a limit of detection for the target compounds at 50 ppb (v/v) and a linear range from 0.5 to 50 ppm with a correlation coefficient of at least 0.997.

Characterization of phenol and alkyl phenols in organic matrixes with monoethylene glycol extraction and multidimensional gas chromatography/mass spectrometry.

The use of monoethylene glycol as an extraction medium for removing phenol and alkyl phenols in organic matrixes such as hydrocarbons is introduced and combined with a practical analytical multidimensional gas chromatography approach. The analytical approach has been successfully developed for the characterization of phenol, cresols, xylenols, and alkyl phenols like 4-ethylphenol and 2,3,5-trimethylphenol. The technique employs a single-step extraction of the analytes with monoethylene glycol and sonication, followed by multidimensional gas chromatography with mass spectrometry in selected ion monitoring mode for the detection and quantitation. Extraction efficiency of phenol approached 100% while cresols, xylenols, and 4-ethylphenol were 97% or higher and 2,3,5-trimethylphenol was better than 91% under the analytical conditions used. With the technique described, a complete analysis can be conducted in less than 16 min. Reproducibility of area counts at two levels, namely, 5 ppm(w) and 50 ppm(w) over a period of 2 days were found to be less than 4% (n = 20). The analytes of interest was found to be linear over a range from 100 ppb(w) to 250 ppm(w) with correlation coefficient of at least 0.999 and detection limit of 50 ppb(w) . Spike recoveries from 500 ppb(w) to 250 ppm(w) for all analytes range from 96 to 102%.

Assessment of asphalt concrete acoustic performance in urban streets.

Geo-referenced close proximity rolling noise and sound absorption measurements are used for acoustical characterization of asphalt concrete surfaces in an urban environment. A close proximity noise map of streets with low speed limits is presented for a reference speed of 50 km/h. Different pavements and pavement conditions, common in urban streets, are analyzed: dense and semidense asphalt concrete, with Spanish denomination D-8 and S-12, respectively, and on the other hand, dense pavement at the end of its service life (D-8(*)). From the acoustics point of view, the most favorable surface, by more than 4 dB(A) compared with the S-12 mix, is the smoothest surface, i.e., the D-8 mix, even though it presents a minor absorption coefficient in normal incidence. Noise levels from dense surfaces (D-8) increase significantly over time, principally due to the appearance of surface defects such as cracks and ruts. Longitudinal variability of the close proximity tire/pavement noise emission and surface homogeneity are also analyzed.

Capillary flow technology with multi-dimensional gas chromatography for trace analysis of oxygenated compounds in complex hydrocarbon matrices.

By employing multi-dimension gas chromatography with capillary flow technology in combination with highly selective capillary columns and a pressurized liquid injection system, light oxygenated compounds such as methanol, ethanol, n-propanol, 2-propanol, and n-butanol in the presence of either light hydrocarbon, heavy hydrocarbon, or aromatic matrices can be measured accurately with minimal possibility of a false positive. Using this technique, a detection limit of at least 0.20 ppm (w/w) with a linear correlation coefficient greater than 0.9993 over a range from 0.5 ppm to 600 ppm (w/w) and a relative standard deviation of greater than 2.7% are achieved for the solutes tested. The technique can also be effective for the measurement of other classes of oxygenated compounds such as ethers, aldehydes, and ketones. Another added benefit for the implementation of capillary flow technology is the capability to conduct column back-flushing, where heavier, undesired solutes in a sample can be back-flushed from the chromatographic system to improve system cleanliness and sample throughput.

Adeno-associated viral serotypes differentially transduce inhibitory neurons within the rat amygdala.

Recombinant adeno-associated viruses (AAV) are frequently used to make localized genetic manipulations within the rodent brain. It is accepted that the different viral serotypes possess differing affinities for particular cell types, but it is not clear how these properties affect their ability to transduce specific neuronal cell sub-types. Here, we examined ten AAV serotypes for their ability to transduce neurons within the rat basal and lateral nuclei of the amygdala (BLA) and the central nucleus of the amygdala (CeA). AAV2 based viral genomes designed to express either green fluorescent protein (GFP) from a glutamate decarboxylase (GAD65) promoter or the far-red fluorescent protein (E2-Crimson) from a phosphate-activated glutaminase (PAG) promoter were created and pseudotyped as AAV2/1, AAV2/4, AAV2/5, AAV2/6, AAV2/7, AAV 2/8, AAV2/9, AAV2/rh10, AAV2/DJ and AAV2/DJ8. These viruses were infused into the BLA and CeA at equal titers and twenty-one days later tissue within the amygdala was examined for viral transduction efficiency. These serotypes transduced neurons with similar efficiency, except for AAV4 and AAV5, which exhibited significantly less efficient neuronal transduction. Notably, AAV4 and AAV5 possess the most divergent capsid protein sequences compared to the other commonly available serotypes. We found that the Gad65-GFP virus did not exclusively express GFP within inhibitory neurons, as assessed by fluorescent in situ hybridization (FISH), but when this virus was used to transduce CeA neurons, the majority of the neurons that expressed GFP were in fact inhibitory neurons and this was likely due to the fact that this nucleus contains a very high percentage of inhibitory neurons.

Assessment of an action against environmental noise: Acoustic durability of a pavement surface with crumb rubber.

Environmental noise is a worldwide problem that has an adverse effect in the quality of life of urban population. Some work has shown that there is a correlation between environmental noise and health issues as sleep disturbance or annoyance. This study presents the time evolution of a test track fabricated with an asphalt mixture with 20% of crumb rubber by weight of bitumen, added by the wet process. A complete surface characterization has been performed by determining tire/pavement sound levels, road texture profiles, in-situ dynamic stiffness and sound absorption of compacted and extracted sample cores. Two measurement campaigns were performed: just after mixture laying and after 3 years in service. This study confirms that the use of crumb rubber as a modifier of bituminous binders (CRMB) can improve the pavement characteristics: gap-graded mixtures with crumb rubber can be used in the action plans as urban rehabilitation measure to fight noise pollution. However, this noise reduction seems to decrease with age at a rate of approximately 0.15 dB(A) per year.

Developments and applications of biosensors in food analysis.

The food industry needs suitable analytical methods for process and quality control; that is, methods that are rapid, reliable, specific and cost-effective in their provision of information about physical and chemical characteristics of food. Apart from a few important analytes, such as sugars, alcohols, amino acids, flavours and sweeteners, food applications mainly focus on the determination of contaminants. However, very few biosensors play a prominent role in food processing or quality control. Considerable effort must be made to develop biosensors that are inexpensive, reliable, and robust enough to operate under realistic conditions.

Enzyme reactions in the presence of cyclodextrins: biosensors and enzyme assays.

Cyclodextrins, macrocyclic carbohydrates with apolar internal cavities, can form complexes with, and solubilize many normally water-insoluble compounds. Ferrocene and its derivatives, tetrathiafulvalene and tetramethylbenzidine, can function as redox mediators, but are insoluble in water; when they are complexed with cyclodextrins, they can be used in enzymatic assays and in the construction of mediated biosensors. In addition, the solubilization of polynuclear aromatic hydrocarbons (PAHs), including the potent carcinogen benzo[a]pyrene, by cyclodextrins has enabled the detection of these important environmental contaminants.

Improvement of the selectivity of an FIA amperometric biosensor system for glucose.

A flow injection analysis (FIA) biosensor system has been developed for the determination of glucose from urine, blood plasma and foodstuffs. Glucose oxidase was immobilized onto porous aminopropyl glass beads via glutaraldehyde activation to form an enzyme column. The hydrogen peroxide released from the conversion of glucose to gluconic acid was monitored by a platinum electrode vs. silver/silver chloride poised at +700 mV. As a novel aspect to the improvement of the selectivity of the biosensor system, an anion exchange column was placed upstream to remove uric acid, ascorbic acid or acetaminophen, three major electroactive interfering substances which usually occur in urine and blood plasma. Among several resins tested, the effective adsorption of uric and ascorbic acids could be accomplished using an acetate anion exchanger, and the selectivity coefficient was pH dependent. The binding of acetaminophen to the resin was much less efficient and, in all cases, the selectivity coefficient was independent of the operating temperature up to 37 degrees C. When applied to real samples, the data obtained by the biosensor system compared well with those of the standard hexokinase assay. The immobilized glucose oxidase could be reused for at least 2000 repeated analyses without loss of its original activity.

An FIA biosensor system for the determination of phosphate.

A flow injection analysis (FIA) biosensor system for the determination of phosphate was constructed using immobilized nucleoside phosphorylase and xanthine oxidase and an amperometric electrode (platinum vs silver/silver chloride, polarized at 0.7 V). When a phosphate-containing sample was injected into the detection cell, phosphate reacted with inosine in the carrier buffer to produce hypoxanthine and ribose-1-phosphate in the presence of nucleoside phosphorylase. Hypoxanthine was then oxidized by xanthine oxidase to uric acid and hydrogen peroxide, which were both detected by the amperometric electrode. The response of the FIA biosensor system was linear up to 100 microM phosphate, with a minimum detectable concentration of 1.25 microM phosphate. Each assay could be performed in 5-6 min and the system could be used for about 160 repeated analyses. This system was applicable for the determination of phosphate in various food products and plasma, and the results obtained agreed well with those of the enzymatic assay.

A microbial biosensor for trimethylamine using Pseudomonas aminovorans cells.

A biosensor system based on the difference in the oxygen uptake response of two microbial electrodes was developed to monitor trimethylamine (TMA). The first electrode, constructed using Pseudomonas aminovorans grown on TMA, was sensitive to TMA, trimethylamine N-oxide (TMAO), dimethylamine (DMA) and monomethylamine (MMA). The second electrode responding to TMAO, DMA and MMA was prepared using Ps. aminovorans grown on TMAO. The difference in oxygen uptake was linearly related to the TMA concentration in the range of 5-26 microM. The minimum detectable level was 2.6 microM and the relative standard deviation was determined to be 14% for 16 repeated analyses. When operated and stored at 30 degrees C, the response of the system was stable for only 2 days. However, when the biosensor system was operated at 30 degrees C but stored overnight at 4 degrees C, the system was stable up to 20 days. The biosensor system was applicable for the determination of TMA in fish tissue extracts and the results compared well with those determined by HPLC.

Monitoring glutamine in mammalian cell cultures using an amperometric biosensor.

An amperometric biosensor has been developed for monitoring glutamine in the pulsed-batch cultivation of murine hybridoma cells. Glutamine oxidase was cross-linked with bovine serum albumin (BSA) via glutaraldehyde activation and deposited on a preactivated nylon membrane. Glutaminase was then immobilized on the protein layer and the resulting membrane was attached to the sensing area of a hydrogen peroxide probe (platinum vs silver/silver chloride polarized at +0.7 V). An orthogonal test was performed to optimize the activity of the membrane for glutamine with respect to the concentrations of glutamate oxidase, BSA, glutaminase and glutaraldehyde. There was an excellent linear relationship between the biosensor's response and glutamine in the range 0.1-3 mM. The determination of glutamine could be performed in 2 min and each membrane was reused for at least 300 consecutive analyses. The data obtained also agreed well with those high-performance liquid chromatography, thus validating the applicability of the biosensor.

A chemiluminescence fiber-optic biosensor system for the determination of glutamine in mammalian cell cultures.

A chemiluminescence fiber-optic biosensor system has been developed for determining glutamine in hybridoma cell cultures producing monoclonal antibodies against viral surface antigens. Glutaminase and glutamate oxidase (GLO) were immobilized onto aminopropyl glass beads via glutaraldehyde activation separately and packed in a column. Two separate columns containing immobilized GLO and catalase were placed upstream to eliminate endogenous glutamate. In the presence of ferricyanide, luminol reacted with hydrogen peroxide released from the enzymatic reactions to produce a chemiluminescence (CL) light signal which was detected and quantitated with a fiber-optic system. In combination with flow injection analysis it was possible to process samples virtually identically, thus avoiding difficulties in reproducing the CL signal. There was an excellent linear relationship between the CL response and standard glutamine concentration in the range 10(-6) to 10(-3) M. A complete analysis could be performed in 2 min including sampling and washing. Each immobilized enzyme column was stable for at least 300 repeated analyses without any loss of activity. When the biosensor system was used for the determination of glutamine in spent mammalian cell cultures, the values obtained compared well with those of high-performance liquid chromatography, thus validating the applicability of the CL fiber-optic system.

The potential role of biosensors in the food and drink industries.

Despite their apparent potential as analytical tools in the food and drink industries, only a few biosensors are used routinely. This article describes the development of biosensors for these sectors and discusses the technical and economic problems of applying this technology to the monitoring of food and drink products.

Monitoring the activity of glucose oxidase during the cultivation of Aspergillus niger using novel amperometric sensor with 1, 1'-dimethylferricinium as a mediator.

1, 1'-dimethylferricinium (DMF+), a deep blue, and stable mediator, was prepared from a water-soluble 1, 1'-dimethylferrocene(DMF):2-hydroxypropyl- beta-cyclodextrin complex via enzymatic oxidation using immobilised bilirubin oxidase. This mediator was superior to other soluble ferrocenes, notably carboxyferrocene, in terms of both solubility (110 mM vs 0.5 mM) and oxidation potential (150 mV vs 300 mV against Ag/AgCl). Although the cyclic voltammogram of DMF+ was electrochemically equivalent to DMF, the use of the former resulted in a significantly lower background current (< 10 nA vs 30 nA). Because of its higher solubility, concentrated stock solutions of DMF+ can be prepared and supplied to the electrode. This is of particular importance when the signal is severely limited by the rate at which the working electrode can oxidase DMF to DMF+. A linear response of current versus units of glucose oxidase (GOD) was obtained up to 0.5 unit/ml. The detection limit was estimated to be 0.03 unit/ml and the response time was 2.5 min or less. The amperometric system was used successfully to follow the GOD activity during the growth of Aspergillus niger a well-known GOD producer. The results obtained correlated well with a standard absorbance-based assay using dichlorophenol-indophenol (DCPIP). The KM of GOD for the glucose in the lysate was measured as 38 mM. A reduced response and higher KM (48 mM) of the cell homogenate, compared to the lysate, illustrated the requirement for the DMF+ and glucose to diffuse across the cell membrane to interact with GOD in whole cells.

Achiral selectivity in cyclodextrin-modified capillary electrophoresis.

Torus-shaped, circular, and hydrophilic cyclodextrins (CD) have been frequently used in capillary electrophoresis (CE) as buffer modifiers to effect chiral separation of enantiomers of drugs and specialty chemicals. Although less common, both neutral and charged cyclodextrins have also been exploited in CE to optimize the achiral separations of peptides, proteins, small molecules and a variety of positional isomers. Nonionic CDs are only useful for separations of net charged analytes through judicious partitioning of such guest molecules into their hydrophobic cavity of the former. However, they can be used with a surfactant for an effective resolution of neutral solutes as a result of a differential partitioning of such solutes in the micellar and the cyclodextrin-modified buffer phase. Ionic cyclodextrins, particularly, negatively charged derivatives with their own electrophoretic mobilities, increase the separation window and enable better resolution of analytes which weakly complex with or are poorly differentiated by neutral cyclodextrins.