Complete Genome Sequence of Flagellimonas sp. Strain HMM57, Isolated from Sedimentary Layers of Crustose Coralline Algae.

Here, we report the genome sequence of Flagellimonas sp. strain HMM57, which was isolated from sedimentary layers of crustose coralline algae in Jeju Island, South Korea. The genome is complete and consists of 4,159,450 bp, with a GC content of 38.5%, 3,616 predicted protein-coding sequences, and 70 RNA genes.

Draft Genome Sequence of Clostridium butyricum Strain 16-3, Isolated from Neonatal Feces.

Here, we report the genome sequence of Clostridium butyricum strain 16-3, which was isolated from infant feces. The genome contains circular contigs of 3,861,515 bp and 769,300 bp, with G+C contents of 28.8% and 28.3%, respectively.

Complete Genome Sequence of Flagellimonas sp. Strain CMM7, Isolated from a Marine Green Alga, Codium minus (Schmidt) Silva.

Here, we report the genome sequence of Flagellimonas sp. strain CMM7, which was isolated from a marine green alga, Codium minus (Schmidt) Silva, in Jeju Island, South Korea. The genome is complete and consists of 4,421,981 bp, with a GC content of 37.5%, 3,942 predicted protein-coding sequences, and 49 RNA genes.

Complete Genome Sequence of Weissella cibaria Strain BM2, Isolated from Korean Kimchi.

Weissella cibaria appears to have broad-spectrum health benefits. Here, we report the genome sequence of Weissella cibaria strain BM2, which was isolated from homemade kimchi; it consists of one circular chromosome of 2,462,443 bp and one plasmid of 11,067 bp. A total of 2,337 coding sequences were predicted, including 2,117 protein-coding sequences and a G+C content of 45.06%.

Complete Genome Sequence of Bacillus velezensis Strain DKU_NT_04, Isolated from a Traditional Korean Food Made from Soybeans (Cheonggukjang).

In the present work, we report the complete genome sequence of Bacillus velezensis DKU_NT_04, isolated from cheonggukjang, which is a traditional Korean fermented soybean paste. The final genome assembly consists of a 4.328-Mbp chromosome with 4,134 coding sequences and a G+C content of 45.21%.

Complete Genome Sequence of Clostridium butyricum Strain DKU_butyricum 4-1, Isolated from Infant Feces.

Clostridium butyricum is a strictly anaerobic spore-forming bacillus that is commonly present in the gut of humans. We report here the complete genome sequence of Clostridium butyricum strain DKU_butyricum 4-1, isolated from infant feces.

Complete Genome Sequence of Bacillus subtilis Strain DKU_NT_02, Isolated from Traditional Korean Food Using Soybean (Chung-gook-jang) for High-Quality Poly-γ-Glutamic Acid Activity.

The complete genome sequence of Bacillus subtilis strain DKU_NT_02, isolated from traditional Korean food using soybeans (chung-gook-jang), is presented here. This strain was chosen to help identify genetic factors with high-quality poly-γ-glutamic acid (γPGA) activity.

Complete Genome Sequence of Bacillus subtilis Strain DKU_NT_03, Isolated from a Traditional Korean Food Using Soybean (Chung-gook-jang) for High-Quality Nattokinase Activity.

We present here the complete genome sequence of Bacillus subtilis strain DKU_NT_03 isolated from the traditional Korean food chung-gook-jang, which is made from soybeans. This strain was chosen to identify genetic factors with high-quality nattokinase activity.

Draft Genome Sequences of Bacillus subtilis Strain DKU_NT_01 Isolated from Traditional Korean Food Containing Soybean (Chung-gook-jang).

Here, we report the whole-genome sequence of Bacillus subtilis strain DKU_NT_01 isolated from traditional Korean food containing soybean (chung-gook-jang). The de novo genome of Bacillus subtilis strain DKU_NT_01 has one contig and G+C content of 55.4%, is 4,954,264 bp in length, and contains 5,011 coding sequences (CDSs).

Enhancement of cytotoxic effect on human head and neck cancer cells by combination of photodynamic therapy and sulforaphane.

Photodynamic therapy (PDT) is a method to treat cancers using photosensitizer and light. PDT has been tried for several tumors. However, the clinical applications are limited by the toxicity of photosensitizer and narrow effect. Sulforaphane (SFN) is a material of isothiocyanate group and known to have anticancer effect. We evaluated the cytotoxic effect of PDT combined with SFN on human head and neck cancer cells. We measured the cell viability, extent of apoptosis and necrosis, reactive oxygen species (ROS) generation and caspase activation. Cell viability was decreased significantly by combination treatment. Cellular apoptosis and necrosis were increased in combination treatment compared to SFN or PDT. ROS generation was also higher in combination treatment than single treatment. In combination treatment group, apoptosis and necrosis were decreased by administration of sodium azide (SA) which is scavenger of ROS. Increased caspase activation in combination treatment was also inhibited by SA. Combination of PDT and SFN led to enhanced cytotoxic effect on head and neck cancer cells. Combination treatment promoted the ROS generation, which induced cell death through activation of caspase pathway.

A 635-nm light-emitting diode (LED) therapy inhibits bone resorptive osteoclast formation by regulating the actin cytoskeleton.

Bone diseases such as osteoporosis are mainly caused by upregulated activity of osteoclasts. The present study was designed to examine the effects of light-emitting diode (LED) irradiation on the formation and activity of multinucleated osteoclasts, specifically "round-shaped" osteoclast cells (ROC) in different cell types derived from mouse. After 635-nm LED irradiation, the cell viability was evaluated by MTT assay. The amount of total tartrate-resistant acid phosphatase (TRAP) + osteoclast and the number of ROC cells were also estimated by TRAP solution assay and TRAP staining, respectively. Actin rings were stained with rhodamine-conjugated phalloidin, and resorption assay was performed by dentin slices. In addition, gene expression levels between the control and irradiation groups were evaluated by RT-PCR. In a morphological analysis, the formation of ROC was significantly inhibited by 635-nm LED irradiation in the different cell types. Actin rings were seen at cell peripheries in most ROC cells of the control group, but patches containing disorganized actin were found in the irradiation group. Both the number of ROCs and bone resorption activity were much lower in the irradiation group than in the control group. Also, the gene expression levels involved in actin ring formation such as integrin ß3 and c-Src decreased in RT-PCR analysis. Overall, 635-nm LED therapy may play a pivotal role in regulating bone remodeling, and it may prove to be a valuable tool to prevent bone loss in osteoporosis and other resorptive bone diseases.

Antiplatelet activity of L-sulforaphane by regulation of platelet activation factors, glycoprotein IIb/IIIa and thromboxane A2.

L-sulforaphane was identified as an anticarcinogen that could produce quinine reductase and a phase II detoxification enzyme. In recent decades, multi-effects of L-sulforaphane may have been investigated, but, to the authors' knowledge, the antiplatelet activation of L-sulforaphane has not been studied yet.In this study, 2 µg/ml of collagen, 50 µg/ml of ADP and 5 µg/ml of thrombin were used for platelet aggregations with or without L-sulforaphane. L-sulforaphane inhibited the platelet aggregation dose-dependently. Among these platelet activators, collagen was most inhibited by L-sulforaphane, which markedly decreased collagen-induced glycoprotein IIb/IIIa activation and thromboxane A2 (TxA2) formation in vitro. L-sulforaphane also reduced the collagen and epinephrine-induced pulmonary embolism, but did not affect prothrombin time (PT) in vivo. This finding demonstrated that L-sulforaphane inhibited the platelet activation through an intrinsic pathway.L-sulforaphane had a beneficial effect on various pathophysiological pathways of the collagen-induced platelet aggregation and thrombus formation as a selective inhibition of cyclooxygenase and glycoprotein IIb/IIIa antagonist. Thus, we recommend L-sulforaphane as a potential antithrombotic drug.

Combination treatment with photodynamic therapy and curcumin induces mitochondria-dependent apoptosis in AMC-HN3 cells.

Photodynamic therapy (PDT) is a treatment for the selective destruction of cancerous and non-neoplastic cells that involves the simultaneous presence of light, oxygen and a light-activatable chemical known as a photosensitizer. Curcumin is one of the most extensively investigated phytochemicals with chemopreventive potential and antitumor effects. In this study, the effect of a combination of PDT and curcumin on apoptotic cell death in AMC-HN3 cells and the molecular mechanism underlying apoptosis was examined to confirm the interaction between photofrin-induced PDT and curcumin during combined mortality. The combination treatment with curcumin and PDT inhibited approximately 70% of the cell viability after PDT, whereas the PDT and curcumin only groups showed a 50 and 10% decrease in cell viability, respectively. In addition, the combination treatment increased the apoptotic events, such as nuclear fragmentation and nuclear condensation. This combination group showed an increase in ROS generation that was higher than that observed after each single treatment. Compared to the single agent treatments, the combination therapy induced the enhanced loss of ∆ψm. Furthermore, the cytosolic levels of cytochrome c were significantly elevated in the combination group. Caspases-9, -3 and PARP, which are apoptosis-related proteins induced by mitochondrial activation, were upregulated remarkably by the combination treatment. When co-treated with glutathione, a singlet oxygen quencher, the combination treatment-induced synergistic cytotoxic and apoptotic effects, enhanced the generation of ROS and suppressed the upregulation of caspase-3 and PARP. These results suggest that the combination modality with PDT and curcumin have a better treatment effect in vitro. The induction of mitochondrial-dependent apoptosis due to the increased generation of ROS may be involved in this combination treatment.

Protective Effect of Minocycline Against Cisplatin-induced Ototoxicity.

OBJECTIVES: Cisplatin, a widely used chemotherapeutic agent, has serious side effects, including nephrotoxicity and ototoxicity. Minocycline is a semisynthetic second-generation tetracycline that exerts anti-inflammatory and neuroprotective effects. The purpose of this study was to elucidate the protective effect of minocycline against cisplatin-induced ototoxicity in the auditory hair cell. METHODS: The House Ear Institute-Organ of Corti 1 (HEI-OC1) cell line and guinea pigs were used for in vitro and in vivo experiments. Cells were exposed to cisplatin with or without pre-treatment with minocycline. Cell survival was analyzed using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide). Whole-cell lysates were collected and immunoblotted with antibodies against Bcl-2, p-c-Jun, active caspase-3, cleaved poly (ADP-ribose) polymerase (PARP), and apoptosis-inducing factor (AIF). The guinea pigs received intraperitoneal injections of cisplatin alone or following minocycline pretreatment. The auditory brainstem response was tested and the cochleae were harvested and evaluated using scanning electron microscopy. RESULTS: Survival significantly increased in cells pretreated with minocycline compared with cells exposed to cisplatin alone. Cisplatin treatment increased the expression of active caspase 3, p-c Jun, PARP, and AIF, and pretreatment with minocycline attenuated this response. In animal study, the threshold shift by cisplatin injection in the auditory brainstem response was less pronounced in animals pretreated with minocycline. Scanning electron microscopy revealed more severe damage to the outer hair cells at the basal and middle turns than the apical turn. CONCLUSION: Minocycline partially protects against cisplatin-induced ototoxicity via both caspase-dependent and independent apoptosis pathways.

An in vitro and in vivo study of combination therapy with Photogem®-mediated photodynamic therapy and cisplatin on mouse cancer cells (CT-26).

OBJECTIVE: This study was designed to evaluate the anticancer effect of cisplatin and photodynamic therapy (PDT) combined in vitro and in vivo. BACKGROUND DATA: PDT, these days, is a promising modality for the treatment of cancer and infections. In order to optimize the treatment, cisplatin is often combined with other chemotherapeutic agents. METHODS: Colon cancer cells were incubated with cisplatin (0.1, 1, and 6 µg/ml), followed by photosensitization with Photogem® and irradiation with a 632 nm diode laser at an energy density of 3.2 J/cm(2). An MTT assay was then used to measure cell viability. For in vivo studies, established tumors were treated with cisplatin (3 mg/kg) alone or with PDT (5 mg/kg of Photogem®, 600 J/cm(2)). The sizes of the tumors were continuously measured to note the effects. RESULTS: The cell viability of the combined therapy group was 19.88 ± 0.41, corresponding to a 9% increase compared with that of the cisplatin- or PDT-only groups. In vivo, the tumors treated with PDT or combination therapy disappeared completely three days after each treatment, but on the 14th day, the recurrence rate was significantly lower in the combination therapy group than in the PDT group. CONCLUSIONS: Combination therapy results in an enhanced anticancer effect, presenting the possibility of minimizing the administration dosage of Photogem® and cisplatin.

Photoactivation of 9-hydroxypheophorbide alpha triggers apoptosis through the reactive oxygen species-mediated mitochondrial pathway and endoplasmic reticulum stress in AMC-HN-3 laryngeal cancer cells.

Photodynamic therapy (PDT) is a promising treatment modality for a variety of cancers. It utilizes light-absorbing compounds combined with direct illumination to generate reactive oxygen species in photosensitizer-targeted tumor cells resulting in the final photodamage of tumors. Recently, we demonstrated that a combination modality of 9-hydroxypheophorbide alpha (9-HPbD)-based PDT and carboplatin exerts enhanced cytotoxic and apoptotic effects on AMC-HN-3 laryngeal cancer cells. The present study aimed to investigate the potential apoptotic pathways initiated by 9-HPbD-PDT-mediated reactive oxygen species (ROS) in AMC-HN-3 cells. Cytotoxicity and apoptosis induced by 9-HPbD-PDT were exhibited in a ROS-dependent manner. Mitochondria and the endoplasmic reticulum (ER) were observed as preferential sites of 9-HPbD accumulation in AMC-HN-3 cells. ROS induced by 9-HPbD-PDT directly led to downregulated expression of Bcl-2, loss of mitochondrial membrane potential, release of cytochrome c from mitochondria, elevation of intracellular calcium due to ER stress, as well as induction of CHOP and activation of caspase-3, -8, -9 and -12. Our results demonstrated that ROS induced by 9-HPbD-PDT play a causative role in triggering mitochondrial events, ER stress and probable involvement of the extrinsic apoptotic pathway in AMC-HN-3 cells.

Combination treatment of Cetuximab and photodynamic therapy in SNU-1041 squamous cancer cell line.

Cetuximab (Erbitux) has been highlighted for its anti-proliferative effects in solid tumors and it is currently used as an adjuvant modality with other anti-cancer treatments. Photodynamic therapy (PDT) is used widely in many specialties of medicine. This study evaluated the efficacy of a combination treatment of two modalities (Cetuximab, PDT) both in vivo and in vitro. The SNU-1041 cell line was used for both the in vitro and in vivo studies. The in vivo and in vitro experiments were each classified into four groups, control group, Cetuximab applied group, PDT applied group and combined modality group. A migration study was performed to determine the anti-migration effect of Cetuximab, and a MTT assay was performed to compare the anti-proliferative effect of the modalities in vitro. For the in vivo study, the cells were implanted into 5-week-old nude mice. The measured volume of the tumor for each group was compared as a function of time. In the migration study, the control group showed a longer migration length than the Cetuximab applied group. In the MTT assay, the combination modality group showed less survival than the uni-modality groups. The measured tumor size after treatment showed that the combination treatment was more effective than the single modalities. PDT and Cetuximab are treatment modalities that target different molecular pathways. A combination of these two treatment modalities was found to more effective than an individual treatment. However, further studies will be needed to determine the optimal dose of the photosensitizer and Cetuximab.

Anti-inflammatory and anticarcinogenic effect of genistein alone or in combination with capsaicin in TPA-treated rat mammary glands or mammary cancer cell line.

The topical application of TPA (12-O-tetradecanoylphorbol-13-acetate) to animal skin or direct treatment of TPA to cell cultures leads to inflammatory responses by enhancing cyclooxygenase 2 (COX-2) expression, and specific COX-2 inhibitors counteract this kind of inflammatory response. Furthermore, suppression of these inflammatory events by dietary-origin chemopreventive agents can provide a potential strategy to control carcinogenesis. In this in vivo study, the mammary glands of mature female rats were treated with TPA, and then the effects of genistein alone or in combination with capsaicin on suppression of inflammatory responses were examined. The combined effects of genistein and capsaicin on COX-2, pJNK, pERK, and pp38 expressions were additive or nonadditive, depending on signals tested. In vitro MCF-7 breast cancer cells, the apoptotic bodies as shown with Hoechst 33342 dye, exhibited a synergistic effect between genistein and capsaicin. The abilities of genistein alone or in combination with capsaicin in inhibiting breast cancer cell proliferation through the modulation of AMPK and COX-2 were tested. AMPK activation by genistein in combination with capsaicin is critical for inhibiting COX-2. We propose that genistein in combination with capsaicin exerts anti-inflammatory and anticarcinogenic properties through the modulation of AMPK and COX-2 and possibly various mitogen-activated protein kinases synergistically or nonsynergistically.

Photodynamic therapy with 9-hydroxypheophorbide alpha on AMC-HN-3 human head and neck cancer cells: induction of apoptosis via photoactivation of mitochondria and endoplasmic reticulum.

Skin phototoxicity is one of the main side effects of photodynamic therapy (PDT). To overcome this problem, some new photosensitizers have been developed with longer absorbance wavelengths and shorter half-life in the body. In this study, we investigated the mechanism of PDT mediated by a new chlorophyll derivative photosensitizer, 9-hydroxypheophorbide alpha (9-HPbD), on AMC-HN-3 cancer cells. Phototoxicity and apoptosis on AMC-HN-3 cells induced by 9-HPbD was exhibited in a time- and dose-dependent manner. Mitochondria and endoplasmic reticulum (ER) were observed as preferential sites of 9-HPbD accumulation. Photoactivation of 9-HPbD-loaded AMC-HN-3 cells led to a rapid generation of reactive oxygen species (ROS) at 30 min, followed by a loss of mitochondrial membrane potential (MMP) at 2 h, translocation of apoptosis-inducing factor (AIF) at 2 h, and the release of cytochrome c at 3 h following PDT. Caspase-12, an important caspase involved in ER-induced apoptosis, and C/EBP homologous protein (CHOP), an ER stress inducible transcription factor, were also upregulated after PDT (3-12 h and 6-12 h, respectively). Subsequently, activation of caspase-9 at 6 h, caspase-3 and PARP at 12 h also occurred in PDT-treated AMC-HN-3 cells. The above observations demonstrate that both mitochondria and ER serve not only as the sites of sensitizer binding, but also the subcellular targets of 9-HPbD-PDT, effective activation of which is responsible for 9-HPbD PDT-induced apoptosis in AMC-HN-3 cells.

Enhanced apoptotic effect of combined modality of 9-hydroxypheophorbide alpha-mediated photodynamic therapy and carboplatin on AMC-HN-3 human head and neck cancer cells.

Photodynamic therapy (PDT) has been developed as an effective treatment for malignant disease. Carboplatin (CBDCA), a less nephrotoxic analog of cisdiamminedichloroplatinum (cisplatin), has been widely used for the treatment of multiple malignancies. In this study, we investigated the cytotoxic and apoptotic effect of combined modality of 9-hydroxypheophorbide alpha (9-HPbD)-mediated PDT and CBDCA on AMC-HN-3 human head and neck cancer cell line in vitro. The attached AMC-HN-3 cells were incubated with CBDCA (0.04 mg/ml) for 24 h at 37 degrees C and followed by photosensitization with 9-HPbD for 6 h and laser irradiation with 670 nm diode laser at an intensity of 2.0 J/cm(2) for activating 9-HPbD for 15 min. Then MTT reduction assay and Hoechst 33342 and propidium iodide (PI) double staining were used respectively to measure the cytotoxicity and nuclear morphology at 24 h after PDT. Expression of caspase-3, -9 and poly(ADP-ribose) polymerase (PARP) was detected at 0, 3, 6 and 12 h after irradiation through Western blotting techniques. Compared with PDT and CBDCA alone groups, there was more cytotoxicity and enhanced apoptotic cell death in combination groups. The peaked expression of cleaved form of caspase-3, -9 and PARP occurred approximately 3 h after PDT. There was stronger expression of cleaved caspase-3, -9 and PARP in combination groups than that in PDT or CBDCA alone groups. This study demonstrates that the combined modality resulted in enhanced apoptotic cell death as well as cytotoxic effect on AMC-HN-3 cells in vitro, which suggests the feasibility of combined modality and the possibility of reducing the effective dosage of 9-HPbD and CBDCA and lowering the side effects on normal cells.