RESUMO
Immunoassay detection of O-pinacolyl methylphosphonic acid (PMPA) employing direct coating of N-2-aminoethyl-O-pinacolyl methylphosphonate (hapten B) on microtiter plates is reported. Coating was achieved by covalently linking hapten B to a glutaraldehyde (GA) polymer network directly bound to the polystyrene (PS) surface of a standard 96-well microtiter plate. 4-(2-(O-Pinacolylmethylphosphoryl amino)ethyl amino)-4-oxobutanoic acid (hapten A)-ovalbumin (OVA) conjugate served as the coating antigen for comparison with direct hapten B-coated plates in the CIELISA format. The developed assay employing direct hapten B coated plates demonstrated enhanced sensitivity with the IC(50) value for PMPA being 0.027 µg mL(-1). The assay could detect PMPA even at the concentration of 0.006 µg mL(-1). The mean recovery of standard PMPA (spiked in water) was found to be 83.7%.
Assuntos
Anticorpos Monoclonais/imunologia , Ensaio de Imunoadsorção Enzimática , Haptenos/química , Haptenos/imunologia , Imunoensaio , Poliestirenos/química , Soman/análogos & derivados , Animais , Ligação Competitiva , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Glutaral/imunologia , Haptenos/metabolismo , Ovalbumina/imunologia , Coelhos , Soman/análiseRESUMO
Chikungunya virus (CHIKV) data from population studies are sparse. During the 2006 epidemic, 509 clinical cases (43% attack rate) were identified in a village survey (West India); laboratory investigations demonstrated normal blood cell counts, elevated acute-phase reactants [erythrocyte sedimentation rate, C-reactive protein and interleukin-6 (IL-6)] and excluded malaria and dengue. Acute CHIKV was characterized by high fever, severe peripheral polyarthralgias, axial myalgias and intense fatigue in over 90% of cases; skin rash (34%) and headache (19%) were uncommon. There were 49% and 62% of survey cases seropositive for IgM (rapid assay) and IgG (immunofluorescence) anti-CHIKV antibodies, respectively. Sixty-five percent of cases recovered within 4 weeks. None of the cases died. Of the population, 4·1% and 1·6% suffered from persistent rheumatic pains, predominantly non-specific, at 1 and 2 years, respectively. Chronic inflammatory arthritis was uncommon (0·3% at 1 year) although serum IL-6 often remained elevated in chronic cases. A larger population study is required to describe post-CHIKV rheumatism and its prognosis.
Assuntos
Infecções por Alphavirus/complicações , Infecções por Alphavirus/epidemiologia , Vírus Chikungunya/patogenicidade , Surtos de Doenças , Dor Musculoesquelética/epidemiologia , Dor Musculoesquelética/etiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Infecções por Alphavirus/virologia , Anticorpos Antivirais/sangue , Febre de Chikungunya , Criança , Pré-Escolar , Doença Crônica , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Índia/epidemiologia , Lactente , Recém-Nascido , Interleucina-6/sangue , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , População Rural , Adulto JovemRESUMO
We synthesize porous polyHIPE networks with silanol functionalities in the polyHIPE backbone. These silanol functionalities are used for covalent bonding with silica aerogels embedded in the polyHIPE. Covalent bonding between silica and polyHIPE networks are confirmed using Fourier-transform infrared spectroscopy and scanning electron microscopy. Silica aerogels covalently bonded with polyHIPE network show macroporous and mesoporous morphologies and possess excellent properties like high bendability, high elasticity, superhydrophobicity (~160°), low density (~0.128 g/cm3), and low thermal conductivity (~0.045 W/m·K). Oil absorption from water/oil mixtures and recovery of the absorbed oil (by squeezing) from flexible silica-polyHIPE networks is studied. The silica-polyHIPE is shown to absorb crude oil ~16-times its own weight and can be reused multiple times after recovery. Hence, such materials are very important for oil spill cleanup applications from aqueous systems.
RESUMO
Three haptens of the organophosphorus (OP) toxicant 'sarin' having different spacer arm were designed and synthesized. Haptens were conjugated with BSA (bovine serum albumin) and ovalbumin (OVA) for raising antibody and coating antigen. High antibody titer with higher specificity was obtained from 4-(4-(isopropoxy(methyl)phosphoryloxy)phenylamino)-4-oxobutanoic acid (hapten B) having reasonable long spacer arm. For the standard curve, an IC(50) (inhibitory concentration) of free antigen was found to be 0.415 µg mL(-1) on the basis of indirect competitive ELISA. The study revealed that heterology in competition inhibition enzyme immunoassay (CIEIA) produced remarkable improvement in the sensitivity and specificity of the assay. Under the optimized conditions, the quantitative working range was found to be 0.19-1.56 µg mL(-1) with a limit of detection (LOD) of 0.05 µg mL(-1). The antibodies showed negligible cross reactivity (CR) with other OP toxicants and pesticides, which makes the assay suitable for the selective detection of sarin.
Assuntos
Ensaio de Imunoadsorção Enzimática/métodos , Imunoconjugados/imunologia , Compostos Organofosforados/análise , Sarina/análise , Animais , Anticorpos/química , Bovinos , Técnicas de Química Analítica , Feminino , Cromatografia Gasosa-Espectrometria de Massas/métodos , Haptenos/química , Imunoconjugados/química , Concentração Inibidora 50 , Modelos Químicos , Ovalbumina/química , Coelhos , Reprodutibilidade dos Testes , Albumina Sérica/química , Espectrofotometria Ultravioleta/métodos , TemperaturaRESUMO
AIM OF THE STUDY: The present study aims to evaluate the in vivo antimalarial potential of the leaf extract of Gomphostemma niveum and two new compounds; named as Gomphostenin (GN-6) and acetyl Gomphostenin (GN-10) isolated and purified from this plant against Plasmodium berghei in mice. MATERIALS AND METHODS: The blood schizontocidal activity was performed in early infection of Plasmodium berghei and also in established infection of Plasmodium berghei. The mice were orally administrated with various doses of water and chloroform extracts of leaves of Gomphostemma niveum (GN-W and GN-C, 300, 400 and 500 mg/kg/day), GN-6 and GN-10 (50, 100, 150 and 200mg/kg/day). Chloroquine (8 mg/kg) used as a positive control, while an equal volume of saline plus 0.5% tween-80 was used as a negative control. RESULTS: GN-W, GN-C and pure compounds (GN-6 and GN-10) produced a dose dependent chemosuppression effect at various dose levels. GN-10 exhibited highest percent of chemosuppression i.e. 92.65% at a dose level of 200mg/kg/day. In case of curative test, the survival period of the parasitized infected mice was significantly prolonged at 200mg/kg dose of GN-10. CONCLUSIONS: The studies have indicated that clerodane class of diterpenes GN-6 and GN-10 certainly holds great promise for malaria control and will be useful in antimalarial chemotherapy.