• BMB Reports
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• v.40 no.1
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• pp.107-112
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• 2007

To study the functioning of HSP70 in Escherichia coli, we selected NtHSP70-2 (AY372070) from among three genomic clones isolated in Nicotiana tabacum. Recombinant NtHSP70-2, containing a hexahistidine tag at the amino-terminus, was constructed, expressed in E. coli, and purified by $Ni^{2+}$ affinity chromatography and Q Sepharose Fast Flow anion exchange chromatography. The expressed fusion protein, $H_6NtHSP70$-2 (hexahistidine-tagged Nicotiana tabacum heat shock protein 70-2), maintained the stability of E. coli proteins up to 90$^{\circ}C$. Measuring the light scattering of luciferase (luc) revealed that NtHSP70-2 prevents the aggregation of luc without ATP during high-temperature stress. In a functional bioassay (1 h at 50$^{\circ}C$) for recombinant $H_6NtHSP70$-2, E. coli cells overexpressing $H_6NtHSP70$-2 survived about seven times longer than those lacking $H_6NtHSP70$-2. After 2 h at 50$^{\circ}C$, only the E. coli overexpressing $H_6NtHSP70$-2 survived under such conditions. Our NtHSP70-2 bioassays, as well as in vitro studies, strongly suggest that HSP70 confers thermo-tolerance to E. coli.

• Advances in Traditional Medicine
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• v.8 no.2
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• pp.171-177
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• 2008

Epigallocatechin 3-gallate (EGCG), one of the major catechins of tea, was isolated from the decaffeinated, crude methanolic extract of Indian green tea (Camellia sinensis L. O. Kuntze) using chromatographic techniques. EGCG was then screened for antidiarrhoeal activity against 30 strains (clinical isolates) of V. cholerae, which is a well known Gram negative bacillus functioning as the pathogen of cholera. V. cholerae strains like V. cholerae 69, 71, 83, 214, 978, 1021, 1315, 1347, 1348, 569B and ATCC 14033 were inhibited by EGCG at a concentration of $25\;{\mu}g/ml$ whereas V. cholerae 10, 522, 976 were even more sensitive, being inhibited at $10\;{\mu}g/ml$ level. However, V. cholerae DN 16, DN 26, 30, 42, 56, 58, 113, 117, 564, 593, 972 and ATCC 14035 were inhibited at $50\;{\mu}g/ml$ level of EGCG. Only four strains were inhibited at $100\;{\mu}g/ml$. In this study the isolated compound was found to be bacteriostatic in its mechanism of action. In the in vivo experiment using the rabbit ileal loop model two different dosages of EGCG ($500\;{\mu}g/ml$ and $1,000\;{\mu}g/ml$) were able to protect the animals when they were challenged with V. cholerae 569B in the ileum.

• Molecules and Cells
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• v.20 no.2
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• pp.297-302
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• 2005

The integrity and proper functioning of telomeres require association of telomeric DNA sequences with specific binding proteins. We have characterized PLP-1, a $PUR{\alpha}$ homolog encoded by F45E4.2, which we previously identified as a candidate double stranded telomere binding protein, by affinity chromatography followed by mass spectrometry. PLP-1 bound double-stranded telomeric DNA in vitro as shown by competition assays. Core binding was provided by the third and fourth nucleotides of the TTAGGC telomeric repeat. This is quite different from the binding sequence of CEH-37, another C. elegans telomere binding protein, suggesting that multiple proteins may bind nematode telomeric DNA simultaneously in vivo.

• Biomedical Science Letters
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• v.14 no.3
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• pp.139-146
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• 2008

The peroxisome proliferator-activated receptor ${\gamma}$ $(PPAR{\gamma})$ plays a central role in adipogenesis and lipid storage. The $(PPAR{\gamma})$ ligands, thiazolidinediones (TZDs), enhance in vitro adipogenesis in several cell types, but the role of the TZDs on in vivo adipogenesis is still poorly understood. To investigate how $PPAR{\gamma}$ ligand troglitazone regulates adipogenesis in female mice, we examined the effects of the troglitazone on adipose tissue mass, morphological changes of adipocytes, and the expression of $PPAR{\gamma}$ target and adipocyte-specific genes in low fat diet-fed female C57BL/6 mice. Administration of troglitazone for 13 weeks did not change body and total white adipose tissue weights compared with control mice. Troglitazone treatment also did not cause a significant decrease in the average size of adipocytes in parametrial adipose tissue although it is reported to increase the number of small adipocytes in male animals. Troglitazone did not affect the mRNA expression of $PPAR{\gamma}$ and its target genes as well as adipocyte-specific genes in parametrial adipose tissue. These results suggest that $PPAR{\gamma}$ does not seem to be associated with adipogenesis in females with functioning ovaries and that its inability to induce adipogenesis may be due to sex-related factors.