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Screening and subcloning of high producer transfectomas using semisolid media and automated colony picker

Dharshanan S, Hung CS

Methods Mol Biol, 2014;1131:105-12.
PMID: 24515462 DOI: 10.1007/978-1-62703-992-5_7

Generation of high-producing clones is a perquisite for achieving recombinant protein yields suitable for biopharmaceutical production. However, in many industrially important cell lines used to produce recombinant proteins such as Chinese hamster ovary, mouse myeloma line (NS0), and hybridomas, only a minority of clones show significantly above-average productivity. Thus, in order to have a reasonable probability of finding rare high-producing clones, a large number of clones need to be screened. Limiting dilution cloning is the most commonly used method, owing to its relative simplicity and low cost. However the use of liquid media in this method makes the selection of monoclonal hybridoma and transfectoma colonies to be labor intensive and time consuming, thus significantly limiting the number of clones that can be feasibly screened. Hence, we describe the use of semisolid media to immobilize clones and a high-throughput, automated colony picker (ClonePix FL) to efficiently isolate monoclonal high-producing clones secreting monoclonal antibodies.
Morphological and biochemical changes of andrographolide-induced cell death in human prostatic adenocarcinoma PC-3 cells

Kim TG, Hwi KK, Hung CS

In Vivo, 2005 May-Jun;19(3):551-7.
PMID: 15875775

Andrographolide was extracted and purified from Andrographis panicula using hexane and water partitioning followed by ethyl acetate extraction and chromatography. It showed selective cytotoxicity to prostate cancer PC-3 cells in vitro. The morphological and biochemical changes induced by the extract in carcinoma PC-3 cell death were studied. In andrographolide-treated cells, evidence of apoptosis such as cell shrinkage and surface microvilli loss after 4-hour treatment and chromatin condensation and fragmentation in H&E-stained cells between 4 to 8 hours after treatment were observed. Under electron microscopy, membrane blebbing and apoptotic bodies formation were seen after 8-hour treatment. Using immunocytochemistry staining and cellular caspase-3 activity assay, andrographolide-treated cells showed considerable caspase-3 activation and caspase-8 in PC-3 cells at 4 and 2 hours after treatment, respectively. This suggests andrographolide-induced cell death was achieved through the apoptotic pathway, via the activation of an extrinsic caspase cascade.
Prevalence of burnout syndrome and its associated factors among doctors in Sabah, Malaysia

Yao LM, Hung CS, Ain SN, Pui E, Lee OC

Psychol Health Med, 2021 Feb 18.
PMID: 33602003 DOI: 10.1080/13548506.2021.1891265

Burnout syndrome is a condition marked by emotional exhaustion, deterioration in motivation and lack of commitment, resulting from long-lasting workplace stress, characterized by exhaustion, negative feelings towards one's work and reduced professional efficacy. Healthcare workers are at particularly high risk for burnout due to high levels of stress. Our objectives were to determine the prevalence of burnout syndrome among doctors in Sabah and factors associated with it. This multi-centre cross-sectional study was carried out among doctors working in five Sabah main hospitals. Data were collected by using self-administered questionnaires consisting of basic socio-demographic information and Copenhagen Burnout Inventory (CBI). Overall, a total of 523 doctors consented to participate in this study. The prevalence of personal burnout, work-related burnout, and patient-related burnout among doctors were 57.1%, 48.8%, and 30.4% respectively. Chi-square analysis showed factors that were associated to personal burnout and work-related burnout, were marital status, duration of service and position in the service. Duration of service was the only factor associated with patient-related burnout. The prevalence of burnout syndrome in Sabah should not to be overlooked and more researches are indicated to look into prevention of burnout syndrome in Malaysian setting.
Fulltext Predictors for Successful Endovascular Intervention in Chronic Carotid Artery Total Occlusion

Chen YH, Leong WS, Lin MS, Huang CC, Hung CS, Li HY, et al.

JACC Cardiovasc Interv, 2016 09 12;9(17):1825-32.
PMID: 27609258 DOI: 10.1016/j.jcin.2016.06.015

OBJECTIVES: This study sought to determine predictors for successful endovascular treatment in patients with chronic carotid artery occlusion (CAO).
BACKGROUND: Endovascular recanalization in patients with chronic CAO has been reported to be feasible, but technically challenging.
METHODS: Endovascular attempts in 138 consecutive chronic CAO patients with impaired ipsilateral hemisphere perfusion were reviewed. We analyzed potential variables including epidemiology, symptomatology, angiographic morphology, and interventional techniques in relation to the technical success.
RESULTS: The technical success rate was 61.6%. Multivariate analysis showed absence of prior neurologic event (odds ratio [OR]: 0.27; 95% confidence interval [CI]: 0.10 to 0.76), nontapered stump (OR: 0.18; 95% CI: 0.05 to 0.67), distal internal carotid artery (ICA) reconstitution via contralateral injection (OR: 0.19; 95% CI: 0.05 to 0.75), and distal ICA reconstitution at communicating or ophthalmic segments (OR:0.12; 95% CI: 0.04 to 0.36) to be independent factors associated with lower technical success. Point scores were assigned proportional to model coefficients, and technical success rates were >80% and <40% in patients with scores of ≤1 and ≥4, respectively. The c-indexes for this score system in predicting technical success was 0.820 (95% CI: 0.748 to 0.892; p < 0.001) with a sensitivity of 84.7% and a specificity of 67.9%.
CONCLUSIONS: Absence of prior neurologic event, nontapered stump, distal ICA reconstitution via contralateral injection, and distal ICA reconstitution at communicating or ophthalmic segments were identified as independent negative predictors for technical success in endovascular recanalization for CAO.
Agreement regarding overcoming hypertension in the Asian Hypertension Society Network 2022

Hoshide S, Yamamoto K, Katsurada K, Yano Y, Nishiyama A, Wang JG, et al.

Hypertens Res, 2023 Jan;46(1):3-8.
PMID: 36229522 DOI: 10.1038/s41440-022-00994-1