• Archives of Plastic Surgery
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• 제39권6호
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• pp.585-592
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• 2012

During the past decade, many studies using platelet-rich plasma (PRP) or adipose-derived stem cells (ASCs) have been conducted in various medical fields, from cardiovascular research to applications for corneal diseases. Nonetheless, there are several limitations of practical applications of PRP and ASCs. Most reports of PRP are anecdotal and few include controls to determine the specific role of PRP. There is little consensus regarding PRP production and characterization. Some have reported the development of an antibody to bovine thrombin, which was the initiator of platelet activation. In the case of ASCs, good manufacturing practices are needed for the production of clinical-grade human stem cells, and in vitro expansion of ASCs requires approval of the Korea Food and Drug Administration, such that considerable expense and time are required. Additionally, some have reported that ASCs could have a potential risk of transformation to malignant cells. Therefore, the authors tried to investigate the latest research on the efficacy and safety of PRP and ASCs and report on the current state and regulation of these stem cell-based therapies.

• BMB Reports
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• 제39권6호
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• pp.737-742
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• 2006

Open reading frame 60 of Bombyx mori nucleopolyhedrovirus (Bm60) is located between 56,673 and 57,479 bp in the BmNPV genome which encodes 268 amino acid residues with predicted molecular weight of 31.0 kDa. Bm60 and its homologues have been identified in 11 completely sequenced lepidopteran NPVs. The transcript of Bm60 was detected by RT-PCR at 18-72 h post-infection (p.i.), while the corresponding protein could be detected at 24-72 h p.i. in BmNPV-infected BmN cells by Western blot analysis using a polyclonal antibody against Bm60. The expression of Bm60 was inhibited in the presence of Ara-c, an inhibitor of viral DNA synthesis. These results together indicated that Bm60 was a late gene. The size of Bm60 product was found to be a 31 kDa in BmNPV-infected BmN cells, consistent with predicted molecular weight. Immuno-fluoresence analysis showed that the Bm60 product was first detected in the cytoplasm at 24 h p.i and also located in nucleus during later infection. In conclusion, the available data suggest that Bm60 is a functional ORF of BmNPV and encodes a 31kDa protein expressed in the later stage of infection cycle.

• BMB Reports
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• 제39권6호
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• pp.671-676
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• 2006

The holoenzyme of protein phosphatase (PP) from tulip petals was purified by using hydrophobic interaction, anion exchange and microcystin affinity chromatography to analyze activity towards p-nitrophenyl phosphate (p-NPP). The catalytic subunit of PP was released from its endogenous regulatory subunits by ethanol precipitation and further purified. Both preparations were characterized by immunological and biochemical approaches to be PP2A. On SDS-PAGE, the final purified holoenzyme preparation showed three protein bands estimated at 38, 65, and 75 kDa while the free catalytic subunit preparation showed only the 38 kDa protein. In both preparations, the 38 kDa protein was identified immunologically as the catalytic subunit of PP2A by using a monoclonal antibody against the PP2A catalytic subunit. The final 623- and 748-fold purified holoenzyme and the free catalytic preparations, respectively, exhibited high sensitivity to inhibition by 1 nM okadaic acid when activity was measured with p-NPP. The holoenzyme displayed higher stimulation in the presence of ammonium sulfate than the free catalytic subunit did by protamine, thereby suggesting different enzymatic behaviors.

• BMB Reports
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• 제29권4호
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• pp.372-379
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• 1996

Components I and II (CI&II) are major phosphoproteins in the frog rod outer segments (ROS) of retina, whose phosphorylation is light- and cyclic nucleotide-dependent. Although it was reported that CI & II could be chemically cross-linked to ${\beta}{\gamma}-subunit$ of transducin (${\beta}{\gamma}_t$), it was not clear whether CI&II physically interact with ${\beta}{\gamma}_t$, under native conditions. CI&II extracted by hypotonic washing fo ROS membranes showed an overlapped migration with ${\beta}{\gamma}_t$, in sucrose density gradient centrifugation. The elution profile of CI&II in the peripheral membrane fractions from gel filtration chromatography also overlapped that of ${\beta}{\gamma}_t$. These hydrodynamic parameters indicate that the native molecular state of CI&II in the peripheral membrane fraction appears to be within a complex, most likely with ${\beta}{\gamma}_t$. CI&II coeluted with ${\beta}{\gamma}_t$, showed no phosphorylation by endogenous kinase which phosphorylates a serine of CI&II in other fractions. The purified CI&II were not able to inhibit trypsin-activated cGMP-phosphodiesterase, and CI&II were not recognized by a monoclonal antibody against the ${\gamma}-subunit$ of transducin, indicating that CI&II are not y-subunit of PDE or transducin. Thus, it is likely that native CI&II, which undergo a light-dependent phosphorylation/dephosphorylation cycle, can associate with ${\beta}{\gamma}$, in frog photoreceptor membranes, and the complex formation has an inhibitory effect on the endogenous phosphorylation of CI&II.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.772-776
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• 2004

Physiological and molecular characteristics of Bacillus thuringiensis SR6 and SR8 were investigated, and phase contrast and electron microscopies revealed that a large rhomboidal crystal protein was present in the sporulating cells. SDS-PAGE and Western blot analyses showed that B. thuringiensis SR8 produced 70 kDa protein much more than other proteins, and that the 70 kDa protein could bind to the antibody of B. thuringiensis subsp. tenebrionis-crystal toxin protein, indicating that the crystal 70 kDa protein has an immunological homology with B. thuringiensis subsp. tenebrionis-crystal toxin protein. The DNA fragment of B. thuringiensis subsp. tenebrionis-toxin gene was detected in B. thuringiensis SR6 and SR8 by using PCR amplification analysis. Furthermore, the insect bioassay showed the insecticidal activity against Colorado potato beetle larvae. Based on the physiological and molecular similarities to B. thuringiensis subsp. tenebrionis, it is suggested that the B. thuringiensis SR6 and SR8 may be mutants of the B. thuringiensis subsp. tenebrionis strain overexpressing the crystal of 70 kDa toxin protein.

• The Plant Pathology Journal
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• 제31권1호
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• pp.41-49
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• 2015

Sweet cherry is an important fruit crop with increasing economical value in Turkey and the world. A number of viruses cause diseases and economical losses in sweet cherry. Prune dwarf virus (PDV), is one of the most common viruses of stone fruits including sweet cherry in the world. In this study, PDV was detected from 316 of 521 sweet cherry samples collected from 142 orchards in 10 districts of Isparta province of Turkey by double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA). The presence of PDV in ELISA positive samples was confirmed in 37 isolates by reverse transcription- polymerase chain reaction (RT-PCR) method. A genomic region of 862 bp containing the coat protein (CP) gene of PDV was re-amplified from 21 selected isolates by RT-PCR. Amplified DNA fragments of these isolates were purified and sequenced for molecular characterization and determining genetic diversity of PDV. Sequence comparisons showed 84-99% to 81-100% sequence identity at nucleotide and amino acid level, respectively, of the CP genes of PDV isolates from Isparta and other parts of the world. Phylogenetic analyses of the CP genes of PDV isolates from different geographical origins and diverse hosts revealed that PDV isolates formed different phylogenetic groups. While isolates were not grouped solely based on their geographical origins or hosts, some association between phylogenetic groups and geographical origins or hosts were observed.

• Parasites, Hosts and Diseases
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• 제39권1호
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• pp.49-56
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• 2001

Excretory/secretory proteins (ESP) from Toxoplasma gondii were analyzed to define the function in the penetration process into host cells. Whole ESP obtained at 37$^{\circ}C$ were composed of 15 bands with molecular mass of 110, 97, 86, 80, 70, 60, 54, 42, 40, 36. 30, 28, 26, 22, and 19 kDa. Five ESP of 86, 80, 42, 36, and 28 kDa were reacted with monoclonal antibodies (mAb). named as Tg386 (microheme), Tg485 (surface membrane), Tg786 (rhoptry), Tg378, and Tg556 (both dense granules), respectively. The ESP was released by a temperature-dependent/-independent manner and all at once whenever ready to pour out except Tg786. Each ESP was not exhausted within the parasite but the amount was limited. Tg786 was released continuously with increment, whereas Tg378 and Tg556 were ceased to release after 3 and 4 hr, Dense granular Tg378 and Tg556 were released spontaneously and constitutively before the entry into host cells also. The entry of T. gondii was inhibited by all the mAbs differentially. And the parasite deprived of ESP was inhibited to enter exponentially up to 90.1%. It is suggested that ESP play an essential function to provide appropriate environment for the entry of the parasite into host cells.

• 한국동물위생학회지
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• 제40권3호
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• pp.187-192
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• 2017

In this report, fifteen monoclonal antibodies (MAbs) against an avian influenza virus (H9N2 subtype) were newly produced and characterized. These MAbs proved to react to the epitopes of nucleocapsid protein (NP), hemagglutinin (HA), neuraminidase (NA) and non-structural protein 1 (NS1) of Korean H9N2 strain, respectively. Two HA-specific MAbs showed the ability to inhibit the hemagglutination activity of H9N2 subtype avian influenza virus when tested by hemagglutination inhibition (HI) assay. All MAbs did not cross-react with other avian-origin viruses (Newcastle disease virus, infectious bursal disease virus, infectious bronchitis virus and avian rotavirus) by immunofluorescence test or enzyme-linked immunosorbent assay. The MAbs produced in this study could be useful as the materials for diagnostics and therapeutics against Korean-lineage H9N2 virus infections.

• The Korean Journal of Physiology
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• 제28권1호
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• pp.91-97
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• 1994

In order to produce a specific bombesin antiserum far very sensitive radioimmunoassay, synthetic $[lys^3]-bombesin$ conjugated to bovine serum albumin was subcutaneously injected into guinea pigs. The conjugation was performed using either carbodiimide or gIutaraldehyde as a coupling agent. The antisera were characterized by analysis of Scatchard and Sips plots. The antiserum LBE 2G/2 raised by repeat injection of the immunogen conjugated with carbodiimide showed the titer of 1 : 188,000, very low cross-reactivity to bombesin-like peptides except bombesin, with high affinity constant $(1.64{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.91). The antiserum LBG 1G/2 produced by repeat injection of the immunogen conjugated with glutaraldehyde possessed the titer of 1 : 43,000, high cross-reactivity to some bombesin-like peptides, high affinity constant $(1.19{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.79). These results indicate that the antiserum LBE 2G/2 is specific only to bombesin and that the antiserum LBG IG/2 binds to some bombesin-like peptides such as alytesin, gastrin releasing peptide and neuromedin C. The antiserum LBE 2G/2 is sufficient for the very sensitive radioimmunoassay of bombesin.

• Fisheries and Aquatic Sciences
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• 제21권4호
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• pp.7.1-7.8
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• 2018

In this study, we isolated and characterized the acid-soluble skin collagen of Pacific bluefin tuna (PBT, Thunnus orientalis). The PBT skin collagen was composed of two ${\alpha}$ chains (${\alpha}1$ and ${\alpha}2$) and one ${\beta}$ chain. The denaturation temperature of PBT collagen was low although it was rich in proline and hydroxyproline. The primary structure of PBT skin collagen was almost identical to that of calf and salmon skin collagen; however, it differed with respect to the epitope recognition of the antibody against salmon type I collagen. These results suggest that the primary structure of skin collagen was highly conserved among animal species, although partial sequences that included the epitope structure differed among collagens.