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The neurotransmitter phenylethylamine (PEA) is highly susceptible to oxidation to produce phenylacetic acid (PA). The fact that PEA and PA are both metabolites of phenylalanine (Phe) in humans makes them important indicators in the diagnosis of phenylketonuria. In this work, three-color whole-cell biosensors were developed to simultaneously detect these analytes (Phe, PEA, and PA). The tyrosine-responsive promoter was used to control the production of green fluorescent protein signals in response to Phe levels. The FeaR regulon was first used to indicate the presence of PEA, whereas the Paa regulon was used for the detection of PA. The combination of three sensor strains together made it possible to semiquantify the three analytes according to unique color outputs without cross-interference. We sought to optimize various modular components (ribosomal binding sites and fluorescent proteins) to ensure the rapid generation of fluorescent signals. Finally, the biosensors were implemented within a microchannel device to reduce sample consumption in point-of-care assays.

Aging is the major risk factor for cardiovascular disease, which is the leading cause of mortality worldwide among aging populations. Cisd2 is a prolongevity gene that mediates lifespan in mammals. Previously, our investigations revealed that a persistently high level of Cisd2 expression in mice is able to prevent age-associated cardiac dysfunction. This study was designed to apply a genetic approach that induces cardiac-specific Cisd2 overexpression (Cisd2 icOE) at a late-life stage, namely a time point immediately preceding the onset of old age, and evaluate the translational potential of this approach. Several discoveries are pinpointed. Firstly, Cisd2 is downregulated in the aging heart. This decrease in Cisd2 leads to cardiac dysfunction and impairs electromechanical performance. Intriguingly, Cisd2 icOE prevents an exacerbation of age-associated electromechanical dysfunction. Secondly, Cisd2 icOE ameliorates cardiac fibrosis and improves the integrity of the intercalated discs, thereby reversing various structural abnormalities. Finally, Cisd2 icOE reverses the transcriptomic profile of the aging heart, changing it from an older-age pattern to a younger pattern. Intriguingly, Cisd2 icOE modulates a number of aging-related pathways, namely the sirtuin signaling, autophagy, and senescence pathways, to bring about rejuvenation of the heart as it enters old age. Our findings highlight Cisd2 as a novel molecular target for developing therapies targeting cardiac aging.

Toll-like receptors (TLRs) play crucial roles in host immune defenses. Recently, TLR-mediated autophagy is reported to promote immune responses via increasing antigen processing and presentation in antigen presenting cells. The present study examined whether the synthetic TLR4 activator (CCL-34) could induce autophagy to promote innate and adaptive immunity. In addition, the potential of CCL-34 as an immune adjuvant in vivo was also investigated. Our data using RAW264.7 cells and bone marrow-derived macrophages showed that CCL-34 induced autophagy through a TLR4-NF-κB pathway. The autophagy-related molecules (Nrf2, p62 and Beclin 1) were activated in RAW264.7 cells and bone marrow-derived macrophages under CCL-34 treatment. CCL-34-stimulated macrophages exhibited significant antigen-processing activity and induced the proliferation of antigen-specific CD4+T cells as well as the production of activated T cell-related cytokines, IL-2 and IFN-γ. Furthermore, CCL-34 immunization in mice induced infiltration of monocytes in the peritoneal cavity and elevation of antigen-specific IgG in the serum. CCL-34 treatment in vivo did not cause toxicity based on serum biochemical profiles. Notably, the antigen-specific responses induced by CCL-34 were attenuated by the autophagy inhibitor, 3-methyladenine. In summary, we demonstrated CCL-34 can induce autophagy to promote antigen-specific immune responses and act as an efficient adjuvant.

Chronic myelogenous leukemia (CML) is clinically treated with imatinib, which inhibits the kinase activity of the Bcr-Abl oncoprotein. However, imatinib resistance remains a common clinical issue. Andrographolide, the major compound of the medicinal plant Andrographis paniculata, was reported to exhibit anticancer activity. In this study, we explored the therapeutic potential of andrographolide and its derivative, NCTU-322, against both imatinib-sensitive and imatinib-resistant human CML cell lines. Both andrographolide and NCTU-322 downregulated the Bcr-Abl oncoprotein in imatinib-resistant CML cells through an Hsp90-dependent mechanism similar to that observed in imatinib-sensitive CML cells. In addition, NCTU-322 had stronger effects than andrographolide on downregulation of Bcr-Abl oncoprotein, induction of Hsp90 cleavage and cytotoxicity of CML cells. Notably, andrographolide and NCTU-322 could induce differentiation, mitotic arrest and apoptosis of both imatinib-sensitive and imatinib-resistant CML cells. Finally, the anticancer activity of NCTU-322 against imatinib-resistant CML cells was demonstrated in vivo. In summary, our data demonstrated that andrographolide and NCTU-322 inhibit Bcr-abl function via a mechanism different from that of imatinib, and they induced multiple anticancer effects in both imatinib-sensitive and resistant CML cells. Our findings demonstrate that andrographolide and NCTU-322 are potential therapeutic agents again CML.

The bacterial strain designated I1-1(T) was isolated from a hot spring located in the Pingtung area, southern Taiwan. Cells of this organism were Gram reaction negative rods, motile by a single polar flagellum. Optimum conditions for growth were 55 degrees C and pH 7. Strain I1-1(T) grew well in lower nutrient media such as 5-10% Luria-Bertani broth, and its extracellular products expressed alkaline protease activity. The 16S rRNA gene sequence analysis indicates that strain I1-1(T) is a member of beta-Proteobacteria. On the basis of a phylogenetic analysis of 16S rDNA sequences, DNA-DNA similarity data, whole-cell protein analysis, physiological and biochemical characteristics, as well as fatty acid compositions, the organism belonged to the genus Tepidimonas and represented a novel species within this genus. The predominant cellular fatty acids of strain I1-1(T) were 16:0 (about 41%), 18:1 omega7c (about 13%), and summed feature 3 [16:1 omega7c or 15:0 iso 2OH or both (about 26%)]. Its DNA base ratio was 68.1 mol%. We propose to classify strain I1-1(T) (=BCRC 17406(T)=LMG 22826(T)) as Tepidimonas taiwanensis sp. nov.

After undergoing keratinase digestion, feather wastes could have a great potential as a source of proteins and amino acids for many applications. In this study, the fermentation conditions of feather-degrading Meiothermus sp. strain I40 (I40) were optimized to enhance the biodegradation of chicken feather to hydrolysates. Initially, the factors essential for I40 keratinase production in submerged fermentation were screened, whereas response surface methodology (RSM) was then employed to evaluate the interactions among the effective factors. At first stage, eight fermentation parameters were screened using a Plackett-Burman (PB) design. Four effective factors identified by PB screening, namely feather concentration, tryptone concentration, yeast extract concentration, and incubation temperature, were further investigated their effects on keratinase production by RSM using central composite design (CCD). The I40 fermentation conditions for maximal keratinase activity were as follows: tryptone concentration 0.16 percent (w/v), yeast extract concentration 0.27 percent (w/v), feather concentration 0.08 percent (w/v), and incubation temperature at 51.7°C for 72 hr under 120 rev/min shaking. Compared to the initial stage, a 13.3-fold increase in keratinase activity was achieved when I40 incubated in the optimized conditions.

A non-pigmented, Gram-negative, aerobic, rod-shaped bacterium, strain GR13(T), was isolated using nutrient agar from a water sample from a pond used for the culture of soft-shell turtles (Trionyx sinensis), Pingtung County, Taiwan. 16S rRNA gene sequence studies indicated that the novel strain formed a monophyletic branch at the periphery of the evolutionary radiation occupied by the genus Vogesella. Its closest neighbours were Vogesella indigofera ATCC 19706(T) and Vogesella perlucida DS-28(T) (both with 97.4 % gene sequence similarity). The novel isolate could be distinguished from these species by several phenotypic characteristics. The predominant fatty acids were summed feature 3 (C(16 : 1)omega7c and/or C(15 : 0) iso 2-OH; 60 %), C(16 : 0) (13.6 %) and C(18 : 1)omega7c (12.5 %). The DNA G+C content of strain GR13(T) was 63 mol%. The DNA-DNA hybridization values for the novel strain with V. indigofera and V. perlucida were25 %. On the basis of the 16S rRNA gene sequence analysis and the chemotaxonomic and physiological data, it is concluded that strain GR13(T) represents a novel species in the genus Vogesella, for which the name Vogesella lacus sp. nov. is proposed. The type strain is GR13(T) (=BCRC 17836(T)=LMG 24504(T)).

A cream-white-coloured, aerobic, Gram-positive, ovoid to spherical-shaped bacterial strain, designated L-143(T), was isolated from soil in Taiwan. The highest 16S rRNA gene sequence similarities of strain L-143(T) (98.3-95.8%) were with members of the genus Rothia. Chemotaxonomic and phenotypic properties of this organism were consistent with its classification in the genus Rothia. The novel isolate was distinguished from all Rothia species by several phenotypic characteristics. The peptidoglycan type was A3alpha, containing lysine, glutamic acid and alanine. The isolate contained MK-7 as the major component of the quinone system. The predominant polar lipid consisted of phosphatidylglycerol and diphosphatidylglycerol, with some unknown phospho- and glycolipids as minor components. The major fatty acids were anteiso-15:0 (57.3%), anteiso-17:0 (17.0%) and 16:0 (9.3%). The G+C content of the genomic DNA was 56.1 mol%. Hence, genotypic, chemotaxonomic and phenotypic data demonstrate that strain L-143(T) should be classified as a novel species in the genus Rothia, for which the name Rothia terrae sp. nov. is proposed. The type strain is L-143(T) (=BCRC 17588(T)=LMG 23708(T)).