• Journal of Hospice and Palliative Care
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• v.24 no.4
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• pp.254-260
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• 2021

Continuous deep sedation (CDS) is an extreme form of palliative sedation to relieve refractory symptoms at the end of life. In this study, we shared our experiences with CDS and examined the clinical characteristics associated with survival in patients with terminal cancer who received CDS. We conducted a chart audit of 106 consecutive patients with terminal cancer who received CDS at a single hospice care unit between January 2014 and December 2016. Survival was defined as the first day of admission to the date of death. The associations between clinical characteristics and survival were presented as hazard ratios and 95% confidence intervals using a Cox proportional hazard model. The mean age of participants was 65.2 years, and 33.0% (n=35) were women. Diazepam was the most commonly administered drug, and haloperidol or lorazepam were also used if needed. One sedative was enough for a majority of the patients. Stepwise multivariate analysis identified poor functioning, a high Palliative Prognostic Index score, hyperbilirubinemia, high serum ferritin levels, and a low number of sedatives as independent poor prognostic factors. Our experiences and findings are expected to be helpful for shared decision-making and further research on palliative sedation.

• Journal of Sericultural and Entomological Science
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• v.35 no.1
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• pp.36-42
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• 1993

Expression of insecticidal protein by fusion product of truncated HD-1[CryIA(a)] N-terminal and HD-73[CryIA(c)] C-Terminal fragment of Bacillus thruingiensis subsp. kurstaki was investigate. Immunological analysis of transformants by using polyclonal antisera raised against the whole-crystal protein of HD-1 revealed that SK4 and SK5 were observed cross-reaction with polypeptides of 77-kDa and 105-kDa, respectively. Bioassay of the transformant pSK5 to Plutella maculipennis and Heliothis assulta were 96% and 97%, respectively.

• Biomolecules & Therapeutics
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• v.9 no.2
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• pp.79-87
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• 2001

Heat shock proteins serve as chaperone by preventing the aggregation of denatured proteins and promote survival of pathogens in harsh environments. In this study, heat shock gene encoding a 84-kDa (p84) protein, which is one of the three major heat shock proteins in S. pneumoniae, was cloned and characterized. PCR with a forward primer derived from N-terminal amino acid sequence of the p84 and a reverse primer derived from the conserved second ATP-binding region of Clp family was used for amplification of the gene encoding the p84 and subsequently the PCR product was used for sequence determination. Sequence analysis of the p84 gene demonstrated that it is a member of ClpL. The deduced amino acid sequence of pneumococcal ClpL shows homology with other members of the Clp family, and particularly, even in variable leader region, with bovine Clp-like protein and L. lactis ClpL. S. pneumoniae clpL is the smallest clop member (701 amono acids) containing the two conserved ATP-binding regions, and hydrophilic N-terminal variable region of pneu-mococcal Clp ATPase is much shorter than any known Clp ATPases. Histidine tagged ClpL was overexpressed and purified from E. coli. Immunoblot analysis employing antisera raised against pneumococcus p84 demonstrated no cross-reactivity with Clp analog in Eschericha coli, Staphylococcus aureus and human HeLa cells. Preimmunization of mice with ClpL extended mice life partially but did not protect them from death.

• BMB Reports
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• v.34 no.4
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• pp.305-309
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• 2001

MatR in Rhizobium trifolii is a malonate-responsive transcription factor that regulates the expression of genes, matABC, enabling decarboxylation of malonyl-CoA into acetyl-CoA, synthesis of malonyl-CoA from malonate and CoA, and malonate transport. According to an analysis of the amino acid sequence homology, MatR belongs to the GntR family The proteins of this family have two-domain folds, the N-terminal helix-turn-helix DNA-binding domain and the C-terminal ligand-binding domain. In order to End the malonate binding site and amino acid residues that interact with RNA polymerase, a site-directed mutagenesis was performed. Analysis of the mutant MatR suggests that Arg-160 might be involved in malonate binding, whereas Arg-102 and Arg-174 are critical for the repression activity by interacting with RNA polymerase.

• Natural Product Sciences
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• v.14 no.3
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• pp.182-186
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• 2008

Scutellaria baicalensis Georgi (SBG) is traditionally used medicinal herb that has anti-oxidant, anticancer and anti-inflammatory effects. In this study, we investigated whether the extracts of SBG have the inhibitory activity in the osteoclast differentiation by using mouse monocytes RAW264.7 cells and primary mouse bone marrow-derived macrophages (BMMs). Methanol extract (ME) from SBG was successively fractionated into methylene chloride (MF), ethylacetate (EF) and n-butanol fraction (BF). The activity assay for tartrateresistant acid phosphatase (TRAP) and Western blot analysis were employed to evaluate the osteoclasts differentiation and the activation of mitogen-activated protein (MAP) kinases, respectively. ME, MF, EF and BF significantly and dose-dependently inhibited osteoclast differentiation without the decrease of cell viability at the concentrations used in this study. In addition, ME significantly inhibited the activation of c-jun-N-terminal kinase (JNK). In conclusion, this study firstly demonstrated that ME of SBG has the potential to inhibit the osteoclast differentiation through the suppression of JNK activation partially.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.54-62
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• 2010

A novel harpin-like protein, HpaXm, was described from cotton leaf blight bacteria, Xanthomonas citri subsp. malvacearum. The hpaXm was found to be localized between hrp2 and hrcC. A phylogenetic analysis of the complete amino acid sequence or solely the 13 highly conserved residues $H_2N$-SEKQLDQLLTQLI-COOH in the N-terminal $\alpha$-helix indicates that HpaXm is evolutionarily closer to HpaGXag and HpaXac than to Hpa1Xoo and Hpa1Xoc. A synthesized peptide containing two heptads, 39-LDQLLTQLIMALLQ-52, from the N-terminal a-helical region of HpaXm displayed comparable activity in inducing a hypersensitive response, but two other synthesized derivatives, $HpaXm{\Delta}T44C$ and $HpaXm{\Delta}M48Q$, showed reduced HR-triggering activity. The data from a GST trap test revealed that HpaXm was released into the extracellular medium, hpaXm mutant deficient for the leader peptide (1-MNSLNTQIGANSSFL-15) was unable to be secreted outside cells but still induced HR in tobacco leaves.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.213-223
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• 2009

Lipid transfer proteins (LTPs) are widely distributed in the plant kingdom, but their functions remain elusive. The proteins AlLTP2-4 were isolated from three related Allium plants: garlic (A. sativum L.), Welsh onion (A. fistulosum L.), and Nanking shallot (A. ascalonicum L.). These novel proteins comprise a new class of LTPs associated with the Ace-AMP1 from onion (A. cepa L.). The AlLTP genes encode proteins harboring 132 common amino acids and also share a high level of sequence identity. Protein characteristics and phylogenetic analysis suggest that LTPs could be classified into five distinct groups. The AlLTPs were clustered into the most distantly related plant LTP subfamily and appeared to be restricted to the Allium species. In particular, the number of amino acids existing between the fourth and fifth Cys residue was suggested as a conserved motif facilitating the categorization of all the LTP-related proteins in the family. Unlike other LTPs, AlLTPs harboring both the putative C-terminal propeptide and N-terminal signal peptide were predicted to be localized to cytoplasmic vacuoles. When a chimeric GFP protein fused with both N-terminal and C-terminal AlLTP2 signal peptides was expressed in rice cells, the fluorescence signal was detected in the endomembrane compartments, thereby confirming that AlLTPs are an unprecedented intracellular type of LTP. Collectively, our present data demonstrate that AlLTPs are a novel type of LTP associated with the Allium species.

• Journal of Microbiology and Biotechnology
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• v.11 no.3
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• pp.508-514
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• 2001

A stuructural gene (ycl) encoding novel yeast cell wall hydrolase, YCL, was cloned from alkalophilic Bacillus alcalophilus subsp. YB380 by PCR, and transformed into E. coli JM83. Based on the N-terminal and internal amino acid sequences of the enzyme, primers were designed for PCr. The positive clone that harbors 1.8 kb of the yeast cell wall hydrolase gene was selected by the colony hybridization method with a PCR fragment as a probe. According to the computer analysis, this gene contained a 400-base-paired N-terminal domain of the enzyme. Based on nucletide homology of the cloned gene, a 850 bp fragment was amplified and the C-terminal domain of the enzyme was sequenced. With a combination of the two sequences, a full nucleotide sequence for YCL was obtained. This gene, ycl, consisted of 1,297 nucleotides with 27 nucleotides with 27 amino acids of signal sequence, 83 redundant amino acids of prosequence, and 265 amino acids of the mature protein. This gene was then cloned into the pJH27 shuttle vector and transformed into the Bacillus subtilis DB104 to express the enzyme. It was confirmed that the expressed cell wall hydrolase that was produced by Bacillus subtilis DB104 was the same as that of the donor strain, by Western blot using polyclonal antibody (IgY) prepared from White Leghorn hen. Purified yeast cell wall hydrolase and expressed recombinant protein showed a single band at the same position in the Western blot analysis.

• Microbiology and Biotechnology Letters
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• v.20 no.6
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• pp.655-662
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• 1992

Pectate lyase produced by recombinant strain containing pectate lyase gene from alkalitolerant Bacillus sp. YA-14 was succesively purified with 257.6 purification folds and a 10.2% yields by the affinity method, eM-cellulose column chromatography followed by gel filtration on Sephadex G-I00 column. The optimal pH and temperature for pectate lyase activity were 10.0 and $60^{\circ}C$, respectively. The enzyme was stable between pH 4.0 and 10.0, and up to $50^{\circ}C$. The molecular weight of this enzyme was estimated to be 43,000 daltons by SDS-PAGE. Amino acid analysis showed that the enzyme contained more polar and basic amino acids, especially serine, glycine and tyrosine, than that of various pectate lyase from other strains. The N-terminal amino acid sequence was Ala-Asp-Leu-Gly-His-Gln-Thr.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1054-1059
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• 2005

An extracellular protease was identified from Bacillus subtilis KCCM 10257 by N-terminal sequencing and mass spectral analysis. The molecular mass of the extracellular protease was estimated to be 28 kDa by SDS-PAGE. Sequencing of the N-terminal of the protease revealed the sequence of A(G,S,R)QXVPYG(A)V(P,L)SQ. The N-terminal sequence exhibited close similarity to the sequence of other proteases from Bacillus sp. A mass list of the monoisotopic peaks in the MALDI-TOF spectrum was searched after peptide fragmentation of the protease. Six peptide sequences exhibiting monoisotopic masses of 1,276.61, 1,513.67, 1,652.81, 1,661.83, 1,252.61, and 1,033.46 were observed from the fragmented protease. These monisotopic masses corresponded to the lytic enzyme L27 from Bacillus subtilis 168, and the Mowse score was found to be 75. A doubly charged Top product (MS) at a m/z of 517.3 exhibiting a molecular mass of 1034.6 was further analyzed by de novo sequencing using a PE Sciex QSTAR Hybrid Quadropole-TOF (MS/MS) mass spectrometer. MS/MS spectra of the Top product (MS) at a m/z of 517.3 obtained from the fragmented peptide mixture of protease with Q-star contained the b-ion series of 114.2, 171.2, 286.2, 357.2, 504.2, 667.4, 830.1, and 887.1 and y-ion series of 147.5, 204.2, 367.2, 530.3, 677.4, 748.4, 863.4, and 920.5. The sequence of analyzed peptide ion was identified as LGDAFYYG from the b- and y-ion series by de novo sequencing and corresponded to the results from the MALDI-TOF spectrum. From these results the extracellular protease from Bacillus subtilis KCCM 10257 was successfully identified with the lytic enzyme L27 from Bacillus subtilis 168.