Induction of Apoptosis and Antitumor Activity of Eel Skin Mucus,Containing Lactose-Binding Molecules,on Human Leukemic K562 Cells

For innate immune defense, lower animals such as fish and amphibian are covered with skin mucus, which acts as both a mechanical and biochemical barrier. Although several mucus sources have been isolated and studied for their biochemical and immunological functions, the precise mechanism(s) of action remains unknown. In the present study, we additionally found the eel skin mucus (ESM) to be a promising candidate for use in anti-tumor therapy. Our results showed that the viability of K562 cells was decreased in a dose-dependent manner by treatment with the isolated ESM. The cleaved forms of caspase-9, caspase-3 and poly adenosine diphosphate-ribose polymerase were increased by ESM. The levels of Bax expression and released cytochrome C were also increased after treatment with ESM. Furthermore, during the ESM mediated-apoptosis, phosphorylation levels of ERK1/2 and p38 but not JNK were increased and cell viabilities of the co-treated cells with ESM and inhibitors of ERK 1/2 or p38 were also increased. In addition, treatment with lactose rescued the ESM-mediated decrease in cell viability, indicating lactose-containing glycans in the leukemia cells acted as a counterpart of the ESM for interaction. Taken together, these results suggest that ESM could induce mitochondria-mediated apoptosis through membrane interaction of the K562 human leukemia cells. To the best of our knowledge, this is the first observation that ESM has anti-tumor activity in human cells.     

Synthesis of hydrophilic copolyesters and the characterization of their moisture-related cooling properties

A series of copolyesters (Co-PETs) containing poly(ethylene glycol) (PEG), 5-sodiumsulfodimethyl isophthalate (DMS), and dimethyl isophthalate (DMI) were synthesized via the conventional two-step melt-polycondensation method. The synthesized Co-PETs were characterized by ¹H-NMR spectroscopy, FT-IR spectroscopy, differential scanning calorimeter (DSC), and thermogravimetric analyzer (TGA). The DSC results showed that the melting temperature (T _m) and the heats of fusion (ΔH _m) of Co-PETs decreased with increasing the DMS content in Co-PET, while the inclusion of PEG did not affect their thermal properties significantly. The water absorption and the water contact angle of the Co-PET films were found to be significantly affected by the DMS content rather than PEG content. The moisture-related cooling properties of the fabric samples made of Co-PET 5 as well as PET were evaluated by using liquid moisture management tester (MMT) and Q _max measurements. The MMT and Q _max results indicated that Co-PET 5 fabric containing DMS 1.0 mol% and PEG 10.0 wt% in Co-PET seemed to be a good candidate for the fabric having durable cooling effects.     

Distribution and characterization of IL-10-secreting cells in lymphoid tissues of PCV2-infected pigs

Distribution and characterization of interlukin-10 (IL-10)-secreting cells in lymphoid tissues of pigs naturally infected with porcine circovirus type 2 (PCV2) were evaluated in accordance with PCV2 antigen detection. After screening a total of 56 pigs showing the symptoms of postweaning multisystemic wasting syndrome (PMWS), 15 pigs were PCV2 positive and 5 pigs, which showed stronger positive signals over multiples tissues were further investigated. This study showed that in PCV2-infected lymphoid tissues, particularly mandibular lymph node, spleen and tonsil, IL-10 expression was mainly localized in T-cell rich areas but rarely in B cell rich areas. IL-10 was highly expressed in bystander cells but rarely in PCV2-infected cells. Elevated IL-10 expression was predominantly associated with T cells, but rarely with B cells or with macrophages. The results of this study provide evidence for the role of IL-10 in chronic PCV2 infection and its relation to PCV2 antigen in affected tissues. Constantly elevated levels of IL-10 lead to immunosuppression in persistent and chronic viral infections. The increased IL-10 expression observed in PCV2 infection in this study suggests that IL-10-mediated immunosuppression may play an important role in the pathogenesis and maintenance of naturally occurring PCV2 infection.     

Biological characteristics and nucleotide sequences of three Korean isolates of Zucchini yellow mosaic virus

Zucchini yellow mosaic virus (ZYMV) is one of the most economically important viruses of cucurbit crops, causing severe mosaic, necrosis, and malformation. Three ZYMV isolates were obtained from pumpkins at Andong (ZYMV-PA), Euiryung (ZYMV-PE), and Suwon (ZYMV-PS), and their biological variability was determined on different hosts, including cucurbit crops as well as other indicator plants. ZYMV-PA caused the most severe symptoms, including severe mosaic, size reduction, and deformation, in oriental melon (Cucumis melo) and cucumber (Cucumis sativus) leaves. In contrast, ZYMV-PE and ZYMV-PS caused mild mosaic symptoms on oriental melon and cucumber. The nucleotide sequences of the genomic RNAs were determined and compared to the sequences of other potyviruses, including ZYMV isolates Reunion Island and TW-TN3. Each ZYMV Isolate had a genome of 9593 nucleotides, excluding the poly(A) tail, and contained 139 and 214 nucleotides in the 5- and 3-untranslated regions, respectively. Each had one large open reading frame encoding a protein of about 351kDa. The nucleotide sequences of ZYMV-PA, ZYMV-PE, and ZYMV-PS were more than 96.0% and the deduced amino acid sequences were more than 98.1% identical. When compared with other ZYMV isolates in a phylogenetic analysis, these three viruses formed a distinct virus clade and were more distantly related to other potyviruses (43.5%–62.8% identity).The nucleotide sequence data reported are available in the GenBank database under the accession numbers AY278998 to AY279000 for ZYMV-PA, ZYMV-PE and ZYMV-PS     

Suppression of cytokine-inducible nitric oxide synthesis during intraperitoneal Meth A tumor growth

Nitric oxide (NO*) synthesis is induced within many tumors. The time course of NO* synthesis was evaluated during intra-peritoneal Meth A fibrosarcoma progression. While increasing macrophage recruitment into ascites was noted, inducible nitric oxide synthase (iNOS) antigen and function peaked between days 3-6 after tumor implantation. The capacity of cells to respond to LPS and IFNgamma stimulation was markedly depressed on day 9 and 11. Cellular proliferation correlated in an inverse fashion with levels of NO* synthesis. Electron paramagnetic resonance spectroscopy and nitrotyrosine immunostaining failed to show accumulation of characteristic target cell lesions induced by NO*. These findings lead us to conclude that NO* production was increasingly suppressed during Meth A tumor progression. Depression of NO* production did not correlate with levels of the inhibitory cytokines TGFbeta and IL-10, but could be partially overcome by addition of sepiapterin (a tetrahydrobiopterin prodrug). Thus, depletion of essential co-factors necessary for iNOS function may contribute to depressed NO* responses during cancer progression.     

Acupoint stimulation using bee venom attenuates formalin-induced pain behavior and spinal cord fos expression in rats

In two previous reports, we have demonstrated that injection of bee venom (BV) into an acupoint produces a significant antinociceptive and anti-inflammatory effect in both a mouse model of visceral nociception and a rat model of chronic arthritis. The present study was designed to evaluate the potential antinociceptive effect of BV pretreatment on formalin-induced pain behavior and it associated spinal cord Fos expression in rats. Adult Sprague-Dawley rats were injected with BV directly into the Zusanli (ST36) acupoint or into an arbitrary non-acupoint located on the back. BV pretreatment into the Zusanli acupoint significantly decreased paw-licking time in the late phase of the formalin test. In contrast, BV injected into a non-acupoint in the back region did not suppress the paw-licking time. In addition, BV pretreatment into the Zusanli acupoint markedly inhibited spinal cord Fos expression induced by formalin injection. These findings indicate that BV pretreatment into the Zusanli acupoint has an antinociceptive effect on formalin-induced pain behavior.     

Molecular epidemiological investigation of foot-and-mouth disease virus in Korea in 2000

The genetic relatedness of 7 Korean type O field strains of foot-and-mouth disease virus (FMDV) in clinical specimens collected from 5 different geographic locations in 2000 was investigated. The sequence of 162 nucleotides (nt 478-639) at the 3' end of the 1D (VP1) genes was determined from amplified cDNA fragments, and subjected to the analysis for the sequence identity/divergence and phylogenetic relationship. The overall nucleotide sequence divergence among the 7 field strains was 0 to 3.8%, suggesting that they are closely related to each other. Phylogenetic analysis with the known Middle East-South Asia (ME-SA) topotype strains showed that the 7 Korean field strains formed two distinct clusters within the same lineage of the ME-SA topotype strains. Cluster 1 consisted of the strains of the primary foci of infection (Paju and Hongseong), and closely related to the strains prevailed in the Far East. Cluster 2 comprised those of subsequently affected regions (Boryeong, Yongin, and Chungju), and was further diverged from the Cluster 1. The result of phylogenetic analysis indicated that the Korean strains may have evolved from a common ancestor of the Pan Asia strains, and that at least 2 phylogenetically clustered variants within the same lineage were prevalent during the epidemic. The potential origin and sources of the virus introduction to Korea were discussed.     

Inhibition of rho-associated kinase reduces MLC20 phosphorylation and contractility of intact myometrium and attenuates agonist-induced Ca2+ sensitization of force of permeabilized rat myometrium

The role of rhoA/rho-associated kinase (ROK) signaling pathways in agonist-induced contraction of the rat myometrium was investigated. We measured the [Ca(2+)](i)-force relationship, phosphorylation of myosin regulatory light chains (MLC(20)) in intact tissue and the Ca(2+)-sensitization of force in permeabilized myometrial cells of rat. In measurements of the relationship between [Ca(2+)](i) and tension in intact tissue, Y-27632, a ROK inhibitor, significantly attenuated the carbachol-induced contraction without changing [Ca (2+)](i). Phosphorylation of MLC(20) was increased by carbachol and this increased phosphorylation was blocked by treatment of tissue with Y-27632. In tension measurements of single hyperpermeable cells, carbachol evoked sustained contraction at constant pCa 6.7 and these agonist-induced contractions were decreased by treatment with Y-27632. These results suggest that activation of a ROK-mediated signaling pathway(s) plays an important role in agonist-induced alterations in MLC(20) phosphorylation and force of rat myometrium.