Antibacterial and synergistic effects of Nardostachytis rhizoma extracts on methicillin-resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) is a serious clinical problem worldwide. Few new drugs are available against MRSA, because it has the ability to acquire resistance to most antibiotics which consequently increases the cost of medication. In the present study, the antibacterial activity of Nardostachytis rhizoma was investigated. The most effective method is to develop antibiotics from the natural products without having any toxic or side effects. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases. The use of two drugs in combination is a good alternative to slow the process of developing drug resistance and to restore the effectiveness of drugs that are no longer prescribed. Combination therapy is the most commonly recommended empirical treatment for bacterial infections in intensive care units, where monotherapy may not be effective against all potential pathogens, and for preventing the emergence of resistant. Five clinical isolates (MRSA) were obtained from five different patients at Wonkwang University Hospital (Iksan, South Korea). The other two strains were S. aureus ATCC 33591 (methicillin-resistant strain) and S. aureus ATCC 25923 (methicillin-susceptible strain). Antibacterial activity (minimal inhibitory concentrations, MICs) was determined by broth dilution method, disc diffusion method, MTT test and checkerboard dilution test. Antimicrobial activity of n-hexane fraction of N. rhizoma was significant. Against the seven strains, the disc diffusion test was in the range of 14 to 18 mm and had a MICs ranging from 31.25 to 125 ìg/ml. FICI values for n-hexane fraction (HFL) of N. rhizome + ampicillin (AM) and HFL + oxacillin (OX) were 0.1875 and 0.078125-0.09375, showing the increase of synergistic effect. When combined together, these antibiotic effects were dramatically increased. These effective combinations could be new promising agents in the management of MRSA and MSSA.Key words: Nardostachytis rhizoma, synergism, antibacterial, methicillin-resistant Staphylococcus aureus (MRSA)

Dysregulation of Parkin-mediated mitophagy in thyroid H\ufcrthle cell tumors

Abnormal accumulation of defective mitochondria is the hallmark of oncocytes, which are frequently observed in thyroid H\ufcrthle cell lesions. Autophagy is an essential cellular catabolic mechanism for the degradation of dysfunctional organelles and has been implicated in several human diseases. It is yet unknown how autophagic turnover of defective mitochondria in H\ufcrthle cell tumors is regulated. We characterized the expression patterns of molecular markers including Beclin1, LC3, PINK1 and Parkin, which are required for autophagy or mitophagy, in human oncocytic lesions of the thyroid. To undertake mechanistic studies, we investigated autophagy and mitophagy using XTC.UC1 cells, the only in vitro model of H\ufcrthle cell tumors. Beclin1 and LC3 were highly expressed in oncocytes of H\ufcrthle cell tumors. XTC.UC1 showed autophagic responses to starvation and rapamycin treatment, whereas they displayed ineffective activation of mitophagy, which is triggered by the coordinated action of PINK1 and Parkin in response to CCCP. This resulted in a decreased turnover of abnormal mitochondria. The mechanisms underlying defective mitophagy and mitochondrial turnover were investigated by genetic analysis of the PARK2 gene in XTC.UC1 and H\ufcrthle cell tumor tissues. XTC.UC1 and several tumors harbored the V380L mutation, resulting in dysfunctional autoubiquitination and decreased E3 ligase activity. Consistently, oncocytes in H\ufcrthle cell tumors displayed comparable expression of PINK1 but decreased Parkin expression in comparison to normal thyrocytes. The introduction of wild-type Parkin sensitized XTC.UC1 to death induced by CCCP. This study provides a possible etiological basis for oncocytic formation in heterogeneous H\ufcrthle cell tumors through insufficient mitophagy leading to ineffective turnover of aberrant mitochondria caused by dysfunctional Parkin-mediated pathways of mitochondria quality control