Abstract A wide variety of cellular mechanisms such as cell division and metabolic processes are maintained by protein-protein interactions (PPIs). The identification of PPI through laboratory techniques is costly, time-consuming, difficult, and challenging. However, computational techniques were generated for PPIs prediction. In alfalfa (Madicago sativa), PPI was predicted among 12 MsMAPKs and 4 MsPP2Cs using a docking approach. For homology modelling, the Swiss model was employed while PROCHECK, ERRAT, and Verify3D were used to validate 3D models. The Ramachandran plots were obtained from PROCHECK which showed value more than 90% (nPP2C1, PP2C1, PP2C, and MSK-3 revealed 92.9%, 94.2%, 92.4%, and 91.1% respectively) for high-quality structures. The HawkDock server and the BIPSPI server were used to analyse protein docking and predict interaction sites, respectively. Our findings demonstrated that MsPP2C docking sites play an important role in the identification and docking of MsMAPKs. The binding free energy ranged from -0.16Kcal/mol to -49.15Kcal/mol for all MsMAPKs and MsPP2Cs, indicating that they interact. Docking site analysis showed that there were 48 pairs of PPIs which indicated that MsPP2Cs can perform a vital role in other signaling pathways. This study found that all MsPP2Cs have docking sites for MsMAPKs, indicating that this method can accurately determine protein-protein interactions.

Um método isocrático de cromatografia líquida de alta resolução de fase reversa (RP-HPLC) foi desenvolvido para determinação de gemifloxacin a granel, em formulações e soro humano a 270 nm. A separação cromatográfica foi adquirida em uma coluna Purospher STAR C18 (250 × 4,6 mm, 5 µm) usando a fase móvel metanol:água (90:10, v/v) ajustada para pH 2,8 com ácido fosfórico 85% em fluxo de 1,5 mL min-1 a temperatura ambiente. As curvas de calibração mostraram-se lineares dentro do intervalo de 5-100 µg mL-1 com um coeficiente de correlação de 0,9998. Os limites de detecção (LOD) e de quantificação (LOQ) foram 0,015 and 0,045 µg mL-1, respectivamente. Os resultados de precisão intra- e inter-corridas e de exatidão foram 98,73-100,12%, que foram correlacionados através do teste t student. Este método foi aplicado para interações in vitro do gemifloxacino com elementos essencial e traço.

An isocratic reversed phase high-performance liquid chromatographic (RP-HPLC) method has been developed for the determination of gemifloxacin in bulk, dosage formulations and human serum at 270 nm. Chromatographic separation was achieved on Purospher STAR C18 (250 × 4.6 mm, 5 µm) column using mobile phase methanol:water (90:10, v/v) adjusted pH 2.8 via phosphoric acid 85% having flow rate of 1.5 mL min-1 at ambient temperature. Calibration curves were linear over range of 5-100 µg mL-1 with a correlation coefficient 0.9998. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.015 and 0.045 µg mL-1, respectively. Intra and inter-run precision and accuracy results were 98.73-100.12% and then correlated through student'st-test. This method was further applied for in vitro interactions of gemifloxacin with essential and trace elements.

Simultaneous determination of moxifloxacin (MOX) and H2-antagonists was first time developed in bulk and formulations. Purospher STAR C18 (250 x 4.6 mm, 5 μm) column was used. The mobile phase (methanol: water: ACN, 60:45:5 v/v/v, pH 2.7) was delivered at a flow rate of 1.0 mL min-1, eluent was monitored at 236, 270 and 310 nm for cimetidine, famotidine and ranitidine, respectively. The proposed method is specific, accurate (98-103%), precise (intra-day and inter-day variation 0.098-1.970%) and linear (r>0.998). The LOD and LOQ were 0.006-0.018 and 0.019-0.005 μg mL-1, respectively. The statistical parameters were applied to verify the results. The method is applicable to routine analysis of formulations and interaction of MOX with H2-antagonist.

An isocratic reversed phase high-performance liquid chromatographic (RP-HPLC) method has been developed for the simultaneous determination of gemifloxacin and diuretics (hydrochlorothiazide and furosemide) in bulk, dosage formulations and human serum at 232 nm. Chromatographic separation was achieved on Purospher Start C18 (250 mm x 4.6 mm, 5 µm) column using mobile phase, methanol: water: acetonitrile (70:25:5 v/v/v) adjusted to pH 3.0 via phosphoric acid 85% having flow rate of 0.8 mL min -1 at room temperature. Calibration curves were linear over range of 0.5-10 µg mL -1 with a correlation coefficient ± 0.999. LOD and LOQ were in the ranges of 0.75-2.56 µg mL -1. Intra and inter-run precision and accuracy results were 98.26 to 100.9.