Search Results (3)

Impedance biosensors are manufactured on glass slides using a semiconductor process to monitor cell growth and cell–drug reactions in real time, and the results are compared with biological assay results to confirm the validity of impedance measurement method. Approximately 10,000 cells per well were cultured for 48 h, after which 6.67 μg/mL puromycin was injected to observe apoptosis over the following 48 h. A frequency sweep from 1 kHz to 1 MHz was performed to determine the optimal frequency range, identifying 367–440 kHz as the most sensitive for detecting impedance changes. Impedance was measured every 10 min for 96 h. Capacitance gradually increased during cell proliferation, while after drug administration, a transient increase occurred within 9 h, followed by a rapid decline, indicating cell death within 24 h. The sensor utilized Electrical Cell–substrate Impedance Sensing (ECIS) to detect real-time changes in cell status without the need for staining or destruction. Comparison with conventional biological assays such as MTS and FACS confirmed that the impedance biosensor provided higher sensitivity and quantitative accuracy in monitoring both cell proliferation and apoptosis. This study demonstrates that the developed biosensor enables label-free, non-invasive, and continuous monitoring of cellular behaviors with acceptable coincidence with 3 different biological assay results. Impedance biosensor presents a promising alternative to conventional biological assays and offers potential applications in drug screening, cytotoxicity evaluation, and real-time biological monitoring. Full article

Among ribosomal proteins essential for protein synthesis, the functions of ribosomal protein L5 (RPL5) and RPL11 still remain unclear to date. Here, the roles of RPL5 and RPL11 were investigated in association with p53/p21 signaling in the antitumor effect of puromycin mainly in HCT116 and H1299 cancer cells. Cell proliferation assays using 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assays and colony formation assays, cell cycle analysis, Reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were performed in cancer cells. Puromycin exerted cytotoxic and anti-proliferative effects in p53 wild-type HCT116 more than in p53 null H1299 cells. Consistently, puromycin increased sub-G1, cleaved Poly (ADP-ribose) polymerase (PARP), activated p53, p21, and Mouse double minute 2 homolog (MDM2), and attenuated expression of c-Myc in HCT116 cells. Notably, puromycin upregulated the expression of RPL5 and RPL11 to directly bind to MDM2 in HCT116 cells. Conversely, deletion of RPL5 and RPL11 blocked the activation of p53, p21, and MDM2 in HCT116 cells. Also, puromycin enhanced the antitumor effect with reactivating p53 and inducing tumor apoptosis (RITA) or doxorubicin in HCT116 cells. These findings suggest that puromycin induces p53-dependent apoptosis via upregulation of RPL5 or RPL11 for binding with MDM2, and so can be used more effectively in p53 wild-type cancers by combination with RITA or doxorubicin. Full article

Chloramphenicol (CAM) is the D-threo isomer of a small molecule, consisting of a p-nitrobenzene ring connected to a dichloroacetyl tail through a 2-amino-1,3-propanediol moiety. CAM displays a broad-spectrum bacteriostatic activity by specifically inhibiting the bacterial protein synthesis. In certain but important cases, it also exhibits bactericidal activity, namely against the three most common causes of meningitis, Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis. Resistance to CAM has been frequently reported and ascribed to a variety of mechanisms. However, the most important concerns that limit its clinical utility relate to side effects such as neurotoxicity and hematologic disorders. In this review, we present previous and current efforts to synthesize CAM derivatives with improved pharmacological properties. In addition, we highlight potentially broader roles of these derivatives in investigating the plasticity of the ribosomal catalytic center, the main target of CAM. Full article