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Using C. elegans as a Model in PKD
Published in Jinghua Hu, Yong Yu, Polycystic Kidney Disease, 2019
This protocol is adapted from previous work.2,27 The assay is done using a stereomicroscope at 50× and a camera mounted to record behaviors. Pick 20 L4 males the day before assay.Prepare small 10 μL OP50 bacterial lawn within 8 h of the assay. We observed sharp declines in tail-chasing behavior if OP50 bacterial lawns were older than 8 h.Use EV preps from him-5, and klp-6; him-5 as positive and negative controls, respectively. Adjust EVs sample concentration to 10 ng total protein/μL. Add 10 μL EV sample or M9 buffer as control to the center of the small OP50 bacterial lawn. Wait 10 min or until the lawn is dry with Petri dish lid open.Place one single young adult male in the center of the EV or M9 buffer (control) spotted lawn.Start to record behavior for 5 min with a capture rate of 1 frame/second. Score reversal movement and tail-chasing (male moving backward in a circle with tail touching male's own head) events from recordings. For each assay, at least 5 males are scored per trial, perform 4 trials to obtain statistical data. Statistics analysis can be done by multiple group comparisons by Kruskal–Wallis test for nonparametric data with Dunn's multiple comparison post hoc test, using GraphPad Prism software.
Green synthesis of ZnO-NPs using endophytic fungal extract of Xylaria arbuscula from Blumea axillaris and its biological applications
Published in Artificial Cells, Nanomedicine, and Biotechnology, 2023
Lavanya Nehru, Gayathri Devi Kandasamy, Vanaraj Sekar, Mohammed Ali Alshehri, Chellasamy Panneerselvam, Abdulrahman Alasmari, Preethi Kathirvel
The agar-disc diffusion assay was used to examine the antibacterial potential of bioinspired ZnONPs. The antibacterial action was studied using eight different types of bacteria: Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa, Proteus mirabilis, Staphylococcus epidermidis, Klebsiella pneumoniae, and Bacillus subtilis. 0.5 OD McFarland bacterial suspensions were prepared, and by using a sterile cotton swab, a homogeneous bacterial lawn was made on the freshly prepared Muller-Hinton agar plates. Whatman filter paper was cut into 6-mm discs and sterilised. Then, 50 µL of varied concentrations (25, 50, 75, 100 µg/mL) of ZnONPs were spotted on the sterile disc under aseptic conditions and placed on the surface of the agar medium. Gentamicin was used as a positive control, while DMSO served as a negative control. After 24 h of incubation at 37 °C, the potential of ZnONPs was determined by measuring the inhibitory zones (in mm). The experiments were repeated three times, and the mean inhibition zones were recorded for each trial [28].
Tea extracts inhibit the attachment of streptococci to oral/dental substrata by reducing hydrogen bonding energies
Published in Biofouling, 2022
Yi Wang, Lakshman P. Samaranayake, Gary A. Dykes
Surface hydrophobicity was determined using contact angle measurements for bacterial lawns and hard surfaces as described by Chia et al. (2011). A bacterial lawn was prepared by filtering 20 ml of bacterial suspension (containing ∼ 108 CFU ml−1) through a membrane filter (pore size: 0.22 μm, pore diameter: 25 mm; Millipore) using negative pressure. The cell-filter was air-dried for 30 min, attached to a glass slide using double sided tape and further dried using a freeze drier (FDU-2100, Eyela, Japan) overnight. A 2 μl drop of liquid was placed onto a hard surface or bacterial lawn using a 10 μl syringe fitted with a needle (Ramé-Hart Inc., USA) and the contact angles were measured using a goniometer (Model 250, Ramé-Hart Inc.) with the aid of DROPimage software (Ramé-Hart Inc.). For each material, ten readings were taken for each of three independently prepared samples.
Co-delivery of isotretinoin and clindamycin by phospholipid-based mixed micellar system confers synergistic effect for treatment of acne vulgaris
Published in Expert Opinion on Drug Delivery, 2021
Gajanand Sharma, Yukhti Yachha, Kanika Thakur, Akanksha Mahajan, Gurjeet Kaur, Bhupinder Singh, Kaisar Raza, OP Katare
Anti-acne susceptibility was performed by cup plate diffusion method. The standard cultures (100 µL) of microbes were poured on top of the agar in the plates for the formation of bacterial lawn [26]. The wells of 4 mm were bored aseptically using sterile cork borer and the holes were filled completely with 100 µL of test solutions. The various formulations tested were viz. Clindamycin phosphate (1%, w/v), ITR (0.05 and 1% w/v in alcohol), ITR + tretinoin (0.05% w/v in alcohol), Tretinoin (1% w/v in alcohol) and phospholipid (1%, w/v) respectively. The plates were kept in an incubator at 37°C for specified time period. Further, petri plates were observed for the antibacterial activity by measuring the zone of inhibition. Clindamycin phosphate solution was used as a standard [43].