13 research outputs found
DEVELOPMENT AND VALIDATION OF STABILITY INDICATING METHOD RP-HPLC METHOD OF ACOTIAMIDE
Objective: The objective of present work was to develop and validate simple, precise, accurate and specific stability indicating method for determination of acotiamide in presence of its degradation products.Methods: An isocratic RP-HPLC method has been developed using C-8 Thermo Hypersil BDS Column (250 x 4.6 mm i.d., 5ĂÂľparticle size) with the mobile phase composition of acetonitrile: 0.1 % triethylamine in 0.2% formic acid (30: 70) at column oven temperature of 40 Ă°C. The flow rate was 1.0 ml min-1 and effluent was detected at 282 nm. The method was validated in terms of linearity, accuracy, precision, LOD (Limit of Detection), LOQ (Limit of Quantification) and robustness as per ICH guidelines.Results: The method was found to be linear in the range of 10-60ĂÂľg/ml. Limit of detection and limit of quantification was found to 0.36ĂÂľg/ml and 1.10 ĂÂľg/ml.% Recovery was found to be in the range of 99.45%-99.75%and precision less than 2%. The developed method was successfully applied for estimation of Acotiamide in marketed tablet formulation and percentage assay was found to be 100.45%. Acotiamide was subjected to stress degradation under acid, base, neutral hydrolysis, oxidation, dry heat, photolysis conditions. Significant degradation was observed in acid and base degradation.Conclusion: The developed RP-HPLC method was simple, rapid, accurate, precise and stability indicating for the estimation of Acotiamide in bulk and tablet dosage form
STABILITY INDICATING HPTLC METHOD FOR DETERMINATION OF CLEVIDIPINE BUTYRATE IN SYNTHETIC MIXTURE
Objective: To develop and validate stability indicating HPTLC method for determination of clevidipine butyrate in synthetic mixture.Methods: The present study deals with development and validation of stability indicating HPTLC method for estimation of clevidipine butryate. Chromatographic separation was performed on aluminum plate pre coated with Silica Gel 60 F254 using toluene: ethyl acetate (8:2) as mobile phase. TLC scanner was set at wavelength of 370 nm.Results: Retention factor Rf of clevidipine was found to be 0.49. The method was validated as per ICH guidelines. Calibration curve was in the range of 1000-6000ng/band. The correlation coefficient was found to be 0.999. The precision expressed by RSD was less than 2%. The accuracy of method was confirmed by recovery studies using standard addition method and recovery was found to be 99.03-99.57%. The drug was subjected to ICH prescribed hydrolytic, oxidative, photolytic and thermal stress conditions. Clevidipine and its degradation products were well resolved under experimental conditions. The method was validated according to ICH guidelines. The drug showed significant degradation in alkaline and acidic condition and slight degradation in oxidative condition. The drug was stable in thermal condition.Conclusion: A new, Simple, Accurate, Precise, Sensitive and economic stability indicating HPTLC method has been developed and validated for the determination of clevidipine and can be employed for stability indicating analysis
Exploration and Profiling of Potential Thermo-alkaliphilic Bacillus licheniformis and Burkholderia sp. from varied Soil of Delhi region, India and their Plant Growth-Promoting Traits
Soilless cultivation has emerged as a fundamental alternative for large-scale vegetable production because it generates high-quality yields and uses resources efficiently. While plant growth-promoting bacteria (PGPB) are known to enhance growth and physiological aspects in crops grown in soil, their application in soilless cultivation has been relatively less explored. This study aimed to isolate potential PGPBs from soil samples collected from five locations in and around the Delhi-National Capital Region (NCR), India, which were further screened for significant PGPB attributes. Among these, 51 isolated were selected for assessing the impact on Oryza sativa (rice) growth and yield grown on a hydroponic set. The results indicated that isolates AFSI16 and ACSI02 significantly improved the physiological parameters of the plants. For instance, treatment with AFSI16 showed a 23.27% increase in maximum fresh shoot mass, while ACSI02 resulted in a 46.8% increase in root fresh mass. Additionally, ACSI02 exhibited the highest shoot length (34.07%), whereas AFSI16 exhibited the longest root length (46.08%) in O.sativa. Treatment with AFSI16 also led to significant increases in total protein content (4.94%) and chlorophyll content (23.44%), while ACSI02 treatment showed a 13.48% increase in maximum carotenoid content in the leaves. The potential PGPBs were identified through 16S rRNA sequencing, as the two most effective strains, AFSI16 and ACSI02, belonged to thermo-alkaliphilic Bacillus licheniformis and Burkholderia sp., respectively. This study demonstrated the potential of these identified PGPB strains in enhancing crop performance, specifically in soilless cultivation systems.
ILâ1β prevents ILC2 expansion, type 2 cytokine secretion, and mucus metaplasia in response to earlyâlife rhinovirus infection in mice
BackgroundEarlyâlife wheezingâassociated respiratory infection with human rhinovirus (RV) is associated with asthma development. RV infection of 6âdayâold immature mice causes mucous metaplasia and airway hyperresponsiveness which is associated with the expansion of ILâ13âproducing type 2 innate lymphoid cells (ILC2s) and dependent on ILâ25 and ILâ33. We examined regulation of this asthmaâlike phenotype by ILâ1β.MethodsSixâdayâold wildâtype or NRLP3â/â mice were inoculated with sham or RVâA1B. Selected mice were treated with ILâ1 receptor antagonist (ILâ1RA), antiâILâ1β, or recombinant ILâ1β.ResultsRhinovirus infection induced Il25, Il33, Il4, Il5, Il13, muc5ac, and gob5 mRNA expression, ILC2 expansion, mucus metaplasia, and airway hyperresponsiveness. RV also induced lung mRNA and protein expression of proâILâ1β and NLRP3 as well as cleavage of caspaseâ1 and proâILâ1β, indicating inflammasome priming and activation. Lung macrophages were a major source of ILâ1β. Inhibition of ILâ1β signaling with ILâ1RA, antiâILâ1β, or NLRP3 KO increased RVâinduced type 2 cytokine immune responses, ILC2 number, and mucus metaplasia, while decreasing ILâ17 mRNA expression. Treatment with ILâ1β had the opposite effect, decreasing ILâ25, ILâ33, and mucous metaplasia while increasing ILâ17 expression. ILâ1β and ILâ17 each suppressed Il25, Il33, and muc5ac mRNA expression in cultured airway epithelial cells. Finally, RVâinfected 6âdayâold mice showed reduced ILâ1β mRNA and protein expression compared to mature mice.ConclusionMacrophage ILâ1β limits type 2 inflammation and mucous metaplasia following RV infection by suppressing epithelial cell innate cytokine expression. Reduced ILâ1β production in immature animals provides a mechanism permitting asthma development after earlyâlife viral infection.Earlyâlife rhinovirus infection increases epithelial expression of the innate cytokines ILâ25 and ILâ33, expands (type 2 innate lymphoid cells) ILC2s, and enhances development of an asthmaâlike phenotype. Rhinovirus causes macrophage (NLR family, pyrin domain containing 3) NLRP3 inflammasome activation and bioactive ILâ1β production. ILâ1β production, which is deficient in immature mice, attenuates production of ILâ25 and ILâ33, thereby protecting against rhinovirusâinduced asthma development.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/156197/3/all14241_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/156197/2/all14241.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/156197/1/all14241-sup-0001-FigS1.pd
Increased expression of miR146a dysregulates TLR2-induced HBD2 in airway epithelial cells from patients with COPD
Background
Airway epithelial cells from patients with COPD show suboptimal innate immune responses to nontypeable Haemophilus influenzae (NTHi) and Toll-like receptor (TLR)2 ligands despite expressing TLR2 similar to normal airway epithelial cells, but the underlying mechanisms are poorly understood.
Methods
Normal or COPD mucociliary-differentiated airway epithelial cells were treated with TLR2 agonists or infected with NTHi and expression of β-defensin (HBD)2 was examined. Interleukin-1 receptor-associated kinase (IRAK)-1 and microRNA (miR)146a were genetically inhibited in normal and COPD airway epithelial cell cultures, respectively, and HBD2 responses to TLR2 ligands were determined. IRAK-1 expression in lung sections was determined by immunofluorescence microscopy.
Results
Compared to normal, COPD airway epithelial cell cultures showed impaired expression of HBD2 in response to TLR2 agonists or NTHi infection. Apical secretions from TLR2 agonist-treated normal, but not COPD, airway epithelial cells efficiently killed NTHi. Knockdown of HBD2 significantly reduced NTHi killing by apical secretions of normal airway epithelial cells. Compared to normal, COPD cells showed significantly reduced expression of IRAK-1 and this was associated with increased expression of miR146a. Inhibition of miR146a increased the expression of IRAK-1, improved the expression of HBD2 in response to TLR2 agonists in COPD cells and enhanced the killing of bacteria by apical secretions obtained from TLR2 agonist-treated COPD cells. Bronchial epithelium of COPD patients showed reduced expression of IRAK-1.
Conclusions
These results suggest that reduced levels of IRAK-1 due to increased expression of miR146a may contribute to impaired expression of TLR2-induced HBD2 in COPD airway epithelial cells
Small Animal Models of Respiratory Viral Infection Related to Asthma
Respiratory viral infections are strongly associated with asthma exacerbations. Rhinovirus is most frequently-detected pathogen; followed by respiratory syncytial virus; metapneumovirus; parainfluenza virus; enterovirus and coronavirus. In addition; viral infection; in combination with genetics; allergen exposure; microbiome and other pathogens; may play a role in asthma development. In particular; asthma development has been linked to wheezing-associated respiratory viral infections in early life. To understand underlying mechanisms of viral-induced airways disease; investigators have studied respiratory viral infections in small animals. This report reviews animal models of human respiratory viral infection employing mice; rats; guinea pigs; hamsters and ferrets. Investigators have modeled asthma exacerbations by infecting mice with allergic airways disease. Asthma development has been modeled by administration of virus to immature animals. Small animal models of respiratory viral infection will identify cell and molecular targets for the treatment of asthma