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Article: Novel Tetra-Primer ARMS-PCR Assays for Thiopurine Intolerance Susceptibility Mutations NUDT15 c.415C>T and TPMT c.719A>G (TPMT*3C) in East Asians.

Ho, Chi-Chun / Fong, Wai-Ying / Lee, Yuen-Hon / Poon, Wing-Tat

2017 Volume 8, Issue 10

Abstract: Thiopurines are clinically useful in the management of diverse immunological and malignant conditions. Nevertheless, these purine analogues can cause lethal myelosuppression, which may be prevented by prospective testing for variants in the thiopurine S- ... ...

Abstract	Thiopurines are clinically useful in the management of diverse immunological and malignant conditions. Nevertheless, these purine analogues can cause lethal myelosuppression, which may be prevented by prospective testing for variants in the thiopurine S-methyltransferase (
Language	English
Publishing date	2017-10-23
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527218-4
ISSN	2073-4425
ISSN	2073-4425
DOI	10.3390/genes8100285
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Article: Comparison of Direct Sequencing, Real-Time PCR-High Resolution Melt (PCR-HRM) and PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis for Genotyping of Common Thiopurine Intolerant Variant Alleles NUDT15 c.415C>T and TPMT c.719A>G (TPMT*3C).

Fong, Wai-Ying / Ho, Chi-Chun / Poon, Wing-Tat

Diagnostics (Basel, Switzerland)

2017 Volume 7, Issue 2

Abstract: ... T allele) and TPMT: NM_000367.4:c.719A>G (TPMT*3C, dbSNP rs1142345 G allele) in East Asians ... NUDT15). Genetic testing of the common variants NUDT15:NM_018283.2:c.415C>T (Arg139Cys, dbSNP rs116855232 ... standard. Sixty patient samples were tested, revealing seven patients (11.7%) heterozygous for NUDT15 c ...

Abstract	Thiopurine intolerance and treatment-related toxicity, such as fatal myelosuppression, is related to non-function genetic variants encoding thiopurine S-methyltransferase (TPMT) and Nudix hydrolase 15 (NUDT15). Genetic testing of the common variants NUDT15:NM_018283.2:c.415C>T (Arg139Cys, dbSNP rs116855232 T allele) and TPMT: NM_000367.4:c.719A>G (TPMT3C, dbSNP rs1142345 G allele) in East Asians including Chinese can potentially prevent treatment-related complications. Two complementary genotyping approaches, real-time PCR-high resolution melt (PCR-HRM) and PCR-restriction fragment length morphism (PCR-RFLP) analysis were evaluated using conventional PCR and Sanger sequencing genotyping as the gold standard. Sixty patient samples were tested, revealing seven patients (11.7%) heterozygous for NUDT15 c.415C>T, one patient homozygous for the variant and one patient heterozygous for the TPMT3C non-function allele. No patient was found to harbor both variants. In total, nine out of 60 (15%) patients tested had genotypic evidence of thiopurine intolerance, which may require dosage adjustment or alternative medication should they be started on azathioprine, mercaptopurine or thioguanine. The two newly developed assays were more efficient and showed complete concordance (60/60, 100%) compared to the Sanger sequencing results. Accurate and cost-effective genotyping assays by real-time PCR-HRM and PCR-RFLP for NUDT15 c.415C>T and TPMT*3C were successfully developed. Further studies may establish their roles in genotype-informed clinical decision-making in the prevention of morbidity and mortality due to thiopurine intolerance.
Language	English
Publishing date	2017-05-12
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662336-5
ISSN	2075-4418
ISSN	2075-4418
DOI	10.3390/diagnostics7020027
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Article ; Online: Structure-based optimization of FDA-approved drug methylene blue as a c-myc G-quadruplex DNA stabilizer.

Chan, Daniel Shiu-Hin / Yang, Hui / Kwan, Maria Hiu-Tung / Cheng, Zhen / Lee, Paul / Bai, Li-Ping / Jiang, Zhi-Hong / Wong, Chun-Yuen / Fong, Wang-Fun / Leung, Chung-Hang / Ma, Dik-Lung

Biochimie

2011 Volume 93, Issue 6, Page(s) 1055–1064

Abstract: ... the c-myc oncogene G-quadruplex DNA. Based on molecular docking analysis of MB to the c-myc G-quadruplex ... with the c-myc G-quadruplex were investigated using the FID assay. The results showed that the methylene blue ... derivatives 6a-c were able to bind to the c-myc G-quadruplex with greater binding affinity compared ...

Abstract	G-quadruplexes are non-canonical DNA secondary structures putatively present in the promoter regions of oncogenes in the human genome. The targeting of promoter G-quadruplex structures to repress oncogene transcription represents a potential anticancer strategy. Here, we have used high-throughput virtual screening to identify FDA-approved drug methylene blue (MB) as a promising scaffold for binding the c-myc oncogene G-quadruplex DNA. Based on molecular docking analysis of MB to the c-myc G-quadruplex, we designed and screened 50 MB derivatives containing side chains that could interact with the G-quadruplex grooves. As a proof-of-concept, the highest-scoring compounds were synthesized and the interactions with the c-myc G-quadruplex were investigated using the FID assay. The results showed that the methylene blue derivatives 6a-c were able to bind to the c-myc G-quadruplex with greater binding affinity compared to the known G-quadruplex binding ligand, crystal violet. The activity of the most potent compound identified from the FID assay, 6b, as an inhibitor for polymerase-drive DNA extension was examined using a PCR-stop assay and compared against that of the parent compound methylene blue. The results of the PCR-stop assay showed that the addition of the side chain improved the activity of the derivatives as an inhibitor compared to the parent compound. The MB derivative 6b was shown to be highly selective towards c-myc G-quadruplex over double-stranded DNA and other biologically relevant G-quadruplexes using UV-visible spectroscopy and mass spectrometry, respectively. The MB derivative 6b could induce or stabilize c-myc G-quadruplex formation in both cell-free and cellular biological models, and displayed higher cytoxicity against human hepatocarcinoma cells compared to the parent compound, MB.
MeSH term(s)	Antineoplastic Agents/chemical synthesis ; Antineoplastic Agents/pharmacology ; Cell Survival/drug effects ; Drug Approval ; G-Quadruplexes/drug effects ; Gene Expression Regulation/drug effects ; Genes, Reporter ; Genes, myc ; HeLa Cells ; Hep G2 Cells ; Humans ; Luciferases/biosynthesis ; Luciferases/genetics ; Methylene Blue/analogs & derivatives ; Methylene Blue/chemical synthesis ; Methylene Blue/chemistry ; Methylene Blue/pharmacology ; Molecular Dynamics Simulation ; Promoter Regions, Genetic ; Structure-Activity Relationship
Chemical Substances	3,7-bis((4-bromophenylethyl)(methyl)amino)phenothiazin-5-ium ; Antineoplastic Agents ; Luciferases (EC 1.13.12.-) ; Methylene Blue (T42P99266K)
Language	English
Publishing date	2011-06
Publishing country	France
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	120345-9
ISSN	1638-6183 ; 0300-9084
ISSN (online)	1638-6183
ISSN	0300-9084
DOI	10.1016/j.biochi.2011.02.013
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Article ; Online: Erratum to: Ungersma SE, Pacheco G, Ho C, Yee SF, Ross J, van Bruggen N, Peale FV Jr, Ross S, Carano RA. Vessel imaging with viable tumor analysis for quantification of tumor angiogenesis. Magn Reson Med 2010;63:1637–1647.

Ungersma, Sharon E / Pacheco, Glenn / Ho, Calvin / Yee, Sharon Fong / Ross, Jed / van Bruggen, Nicholas / Peale, Franklin V / Ross, Sarajane / Carano, Richard A D

Magnetic resonance in medicine

2011 Volume 65, Issue 3, Page(s) 889–899

Abstract: Imaging of tumor microvasculature has become an important tool for studying angiogenesis and monitoring antiangiogenic therapies. Ultrasmall paramagnetic iron oxide contrast agents for indirect imaging of vasculature offer a method for quantitative ... ...

Abstract	Imaging of tumor microvasculature has become an important tool for studying angiogenesis and monitoring antiangiogenic therapies. Ultrasmall paramagnetic iron oxide contrast agents for indirect imaging of vasculature offer a method for quantitative measurements of vascular biomarkers such as vessel size index, blood volume, and vessel density (Q). Here, this technique is validated with direct comparisons to ex vivo micro-computed tomography angiography and histologic vessel measurements, showing significant correlations between in vivo vascular MRI measurements and ex vivo structural vessel measurements. The sensitivity of the MRI vascular parameters is also demonstrated, in combination with a multispectral analysis technique for segmenting tumor tissue to restrict the analysis to viable tumor tissue and exclude regions of necrosis. It is shown that this viable tumor segmentation increases sensitivity for detection of significant effects on blood volume and Q by two antiangiogenic therapeutics [anti-vascular endothelial growth factor (anti-VEGF) and anti-neuropilin-1] on an HM7 colorectal tumor model. Anti-vascular endothelial growth factor reduced blood volume by 36±3% (p<0.0001) and Q by 52±3% (p<0.0001) at 48 h post-treatment; the effects of anti-neuropilin-1 were roughly half as strong with a reduction in blood volume of 18±6% (p<0.05) and a reduction in Q of 33±5% (p<0.05) at 48 h post-treatment.
MeSH term(s)	Angiography ; Animals ; Antineoplastic Agents/therapeutic use ; Colorectal Neoplasms/diagnostic imaging ; Colorectal Neoplasms/drug therapy ; Magnetic Resonance Imaging/methods ; Mice ; Neovascularization, Pathologic/diagnostic imaging ; Sensitivity and Specificity ; Tomography, X-Ray Computed ; Tumor Burden/drug effects
Chemical Substances	Antineoplastic Agents
Language	English
Publishing date	2011-03-23
Publishing country	United States
Document type	Journal Article ; Corrected and Republished Article
ZDB-ID	605774-3
ISSN	1522-2594 ; 0740-3194
ISSN (online)	1522-2594
ISSN	0740-3194
DOI	10.1002/mrm.22880
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Article: Structure-based optimization of FDA-approved drug methylene blue as a c-myc G-quadruplex DNA stabilizer

Chan, Daniel Shiu-Hin / Yang, Hui / Kwan, Maria Hiu-Tung / Cheng, Zhen / Lee, Paul / Bai, Li-Ping / Jiang, Zhi-Hong / Wong, Chun-Yuen / Fong, Wang-Fun / Leung, Chung-Hang / Ma, Dik-Lung

Biochimie. 2011 June, v. 93, no. 6

2011

Abstract: ... the c-myc oncogene G-quadruplex DNA. Based on molecular docking analysis of MB to the c-myc G-quadruplex ... with the c-myc G-quadruplex were investigated using the FID assay. The results showed that the methylene blue ... derivatives 6a–c were able to bind to the c-myc G-quadruplex with greater binding affinity compared ...

Abstract	G-quadruplexes are non-canonical DNA secondary structures putatively present in the promoter regions of oncogenes in the human genome. The targeting of promoter G-quadruplex structures to repress oncogene transcription represents a potential anticancer strategy. Here, we have used high-throughput virtual screening to identify FDA-approved drug methylene blue (MB) as a promising scaffold for binding the c-myc oncogene G-quadruplex DNA. Based on molecular docking analysis of MB to the c-myc G-quadruplex, we designed and screened 50 MB derivatives containing side chains that could interact with the G-quadruplex grooves. As a proof-of-concept, the highest-scoring compounds were synthesized and the interactions with the c-myc G-quadruplex were investigated using the FID assay. The results showed that the methylene blue derivatives 6a–c were able to bind to the c-myc G-quadruplex with greater binding affinity compared to the known G-quadruplex binding ligand, crystal violet. The activity of the most potent compound identified from the FID assay, 6b, as an inhibitor for polymerase-drive DNA extension was examined using a PCR-stop assay and compared against that of the parent compound methylene blue. The results of the PCR-stop assay showed that the addition of the side chain improved the activity of the derivatives as an inhibitor compared to the parent compound. The MB derivative 6b was shown to be highly selective towards c-myc G-quadruplex over double-stranded DNA and other biologically relevant G-quadruplexes using UV–visible spectroscopy and mass spectrometry, respectively. The MB derivative 6b could induce or stabilize c-myc G-quadruplex formation in both cell-free and cellular biological models, and displayed higher cytoxicity against human hepatocarcinoma cells compared to the parent compound, MB.
Keywords	DNA ; binding capacity ; drugs ; gentian violet ; humans ; mass spectrometry ; methylene blue ; molecular models ; oncogenes ; promoter regions ; screening ; transcription (genetics) ; ultraviolet-visible spectroscopy
Language	English
Dates of publication	2011-06
Size	p. 1055-1064.
Publishing place	Elsevier Masson SAS
Document type	Article
ZDB-ID	120345-9
ISSN	0300-9084
ISSN	0300-9084
DOI	10.1016/j.biochi.2011.02.013
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Article: Association between sequence variant of c.553 G > T in the apolipoprotein A5 gene and metabolic syndrome, insulin resistance, and carotid atherosclerosis.

Chien, Kuo-Liong / Hsu, Hsiu-Ching / Chen, Yen-Ching / Su, Ta-Chen / Lee, Yuan-Teh / Chen, Ming-Fong

Translational research : the journal of laboratory and clinical medicine

2009 Volume 154, Issue 3, Page(s) 133–141

Abstract: Common polymorphism of the apolipoprotein A5 gene (APOA5, c.553G>T) related to metabolic syndrome ... of the APOA5 c.553G>T gene with various metabolic syndrome components and carotid artery atherosclerosis ... included in this study. Participants with APOA5 c.553T carrier (GT and TT) were more likely to have higher ...

Abstract	Common polymorphism of the apolipoprotein A5 gene (APOA5, c.553G>T) related to metabolic syndrome components, insulin resistance, and carotid atherosclerosis remains unclear. We investigated the associations of the APOA5 c.553G>T gene with various metabolic syndrome components and carotid artery atherosclerosis among family members. A total of 661 participants who provided complete genotyping and carotid artery measures were included in this study. Participants with APOA5 c.553T carrier (GT and TT) were more likely to have higher levels of triglycerides and apolipoprotein B, as well as lower levels of high-density lipoprotein (HDL) cholesterol, than participants with the GG genotype. Individuals who carried T alleles had an increased risk of a high level of triglycerides (multivariate odds ratio [OR], 3.86; 95% confidence interval [CI], 1.98-7.55; P<0.0001) and low levels of HDL cholesterol (OR, 2.32; 95% CI, 1.40-3.86; P=0.0012) compared with those without T alleles. The age was an effect modifier for the association between APOA5 genotype and smoking, alcohol drinking, obesity, and lipid profiles, including total, HDL, and low-density lipoprotein (LDL) cholesterol; triglycerides; and apolipoproteins. In addition, the association between APOA5 genotype and hypertriglyceridemia was significant only in adult groups (OR, 3.53; 95% CI, 1.79-6.94), and the association between APOA5 genotype and low HDL cholesterol was stable in young adolescents (OR, 2.39; 95% CI, 1.19-4.78) and adults (OR, 2.20; 95% CI, 1.17-4.15). Our findings indicated that the APOA5 c.553G>T polymorphism is associated with high triglycerides and low HDL cholesterol but not with other metabolic syndrome components or carotid atherosclerosis in this ethnic Chinese population.
MeSH term(s)	Adolescent ; Adult ; Apolipoprotein A-V ; Apolipoproteins A/genetics ; Asian Continental Ancestry Group/genetics ; Carotid Artery Diseases/blood ; Carotid Artery Diseases/ethnology ; Carotid Artery Diseases/genetics ; Cholesterol, HDL/blood ; Female ; Genetic Predisposition to Disease/ethnology ; Genetic Variation ; Genotype ; Humans ; Insulin Resistance/ethnology ; Insulin Resistance/genetics ; Male ; Metabolic Syndrome/blood ; Metabolic Syndrome/ethnology ; Metabolic Syndrome/genetics ; Middle Aged ; Risk Factors ; Triglycerides/blood ; Young Adult
Chemical Substances	APOA5 protein, human ; Apolipoprotein A-V ; Apolipoproteins A ; Cholesterol, HDL ; Triglycerides
Language	English
Publishing date	2009-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2246684-8
ISSN	1878-1810 ; 1532-6543 ; 1931-5244
ISSN (online)	1878-1810 ; 1532-6543
ISSN	1931-5244
DOI	10.1016/j.trsl.2009.06.005
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Article: Specific induction of RGS16 (regulator of G-protein signalling 16) mRNA by protein kinase C in CEM leukaemia cells is mediated via tumour necrosis factor alpha in a calcium-sensitive manner.

Fong, C W / Zhang, Y / Neo, S Y / Lin, S C

The Biochemical journal

2000 Volume 352 Pt 3, Page(s) 747–753

Abstract: The RGS (regulator of G-protein signalling) proteins are GTPase-activating proteins for activated ... Galpha subunits. We investigated the effects of protein kinase C (PKC) on RGS proteins in various T ...

Abstract	The RGS (regulator of G-protein signalling) proteins are GTPase-activating proteins for activated Galpha subunits. We investigated the effects of protein kinase C (PKC) on RGS proteins in various T cell lines by treating them with PMA. mRNA levels of both RGS16 and tumour necrosis factor alpha (TNFalpha) were found to be up-regulated in CEM leukaemia cells in a PKC-dependent manner. Mezerein, a non-phorbol-ester activator of PKC, also elevated RGS16 and TNFalpha mRNA levels, while the specific PKC inhibitor Go6983 abrogated their expression. In view of the slower kinetics of PMA-induced RGS16 expression and the tight correlation between TNFalpha and RGS16 mRNA induction among the cell lines studied, we suggest that activation of PKC up-regulates RGS16 via TNFalpha. Indeed, addition of recombinant TNFalpha to CEM cells rapidly stimulated RGS16 mRNA expression independently of PKC. Furthermore, mobilization of calcium by A23187 and thapsigargin blocked the TNFalpha-mediated induction of RGS16, which was reversed by EGTA and by the immunosuppressants FK506 and cyclosporin A, suggesting that the calcineurin/NF-AT (nuclear factor of activated T cells) pathway may repress the up-regulation process. Our results demonstrate for the first time that activation of PKC induces RGS16 expression via TNFalpha in a calcium-sensitive manner, thereby implicating RGS16 in the regulation of T cell responses to inflammation.
MeSH term(s)	Androstadienes/pharmacology ; Bucladesine/pharmacology ; Calcimycin/pharmacology ; Calcineurin/metabolism ; Calcium/antagonists & inhibitors ; Calcium/metabolism ; Calcium Signaling/drug effects ; Cyclosporine/pharmacology ; DNA-Binding Proteins/metabolism ; Egtazic Acid/pharmacology ; Enzyme Activation/drug effects ; Humans ; Kinetics ; Mitogen-Activated Protein Kinases/metabolism ; NFATC Transcription Factors ; Nuclear Proteins ; Protein Kinase C/antagonists & inhibitors ; Protein Kinase C/metabolism ; Proteins/genetics ; RGS Proteins/genetics ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Tacrolimus/pharmacology ; Tetradecanoylphorbol Acetate/pharmacology ; Thapsigargin/pharmacology ; Transcription Factors/metabolism ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha/antagonists & inhibitors ; Tumor Necrosis Factor-alpha/genetics ; Tumor Necrosis Factor-alpha/pharmacology ; Up-Regulation/drug effects
Chemical Substances	Androstadienes ; DNA-Binding Proteins ; NFATC Transcription Factors ; Nuclear Proteins ; Proteins ; RGS Proteins ; RGS16 protein ; RNA, Messenger ; Transcription Factors ; Tumor Necrosis Factor-alpha ; Calcimycin (37H9VM9WZL) ; Egtazic Acid (526U7A2651) ; Bucladesine (63X7MBT2LQ) ; Thapsigargin (67526-95-8) ; Cyclosporine (83HN0GTJ6D) ; Protein Kinase C (EC 2.7.11.13) ; Mitogen-Activated Protein Kinases (EC 2.7.11.24) ; Calcineurin (EC 3.1.3.16) ; Tetradecanoylphorbol Acetate (NI40JAQ945) ; Calcium (SY7Q814VUP) ; Tacrolimus (WM0HAQ4WNM) ; wortmannin (XVA4O219QW)
Language	English
Publishing date	2000-12-15
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2969-5
ISSN	1470-8728 ; 0264-6021 ; 0006-2936 ; 0306-3275
ISSN (online)	1470-8728
ISSN	0264-6021 ; 0006-2936 ; 0306-3275
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Article ; Online: Accurate top protein variant discovery via low-N pick-and-validate machine learning.

Chu, Hoi Yee / Fong, John H C / Thean, Dawn G L / Zhou, Peng / Fung, Frederic K C / Huang, Yuanhua / Wong, Alan S L

Cell systems

2024 Volume 15, Issue 2, Page(s) 193–203.e6

Abstract: A strategy to obtain the greatest number of best-performing variants with least amount of experimental effort over the vast combinatorial mutational landscape would have enormous utility in boosting resource producibility for protein engineering. Toward ... ...

Abstract	A strategy to obtain the greatest number of best-performing variants with least amount of experimental effort over the vast combinatorial mutational landscape would have enormous utility in boosting resource producibility for protein engineering. Toward this goal, we present a simple and effective machine learning-based strategy that outperforms other state-of-the-art methods. Our strategy integrates zero-shot prediction and multi-round sampling to direct active learning via experimenting with only a few predicted top variants. We find that four rounds of low-N pick-and-validate sampling of 12 variants for machine learning yielded the best accuracy of up to 92.6% in selecting the true top 1% variants in combinatorial mutant libraries, whereas two rounds of 24 variants can also be used. We demonstrate our strategy in successfully discovering high-performance protein variants from diverse families including the CRISPR-based genome editors, supporting its generalizable application for solving protein engineering tasks. A record of this paper's transparent peer review process is included in the supplemental information.
MeSH term(s)	Humans ; Mutation/genetics ; Machine Learning ; Protein Engineering ; Genome
Language	English
Publishing date	2024-02-09
Publishing country	United States
Document type	Journal Article
ZDB-ID	2854138-8
ISSN	2405-4720 ; 2405-4712
ISSN (online)	2405-4720
ISSN	2405-4712
DOI	10.1016/j.cels.2024.01.002
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Article ; Online: Home Care Worker Continuity in Home-Based Long-Term Care: Associated Factors and Relationships With Client Health and Well-Being.

Reckrey, Jennifer M / Russell, David / Fong, Mei-Chia / Burgdorf, Julia G / Franzosa, Emily C / Travers, Jasmine L / Ornstein, Katherine A

Innovation in aging

2024 Volume 8, Issue 3, Page(s) igae024

Abstract: Background and objectives: Despite the importance of provider continuity across healthcare settings, continuity among home care workers who provide hands-on long-term care is understudied. This project describes home care worker continuity, identifies ... ...

Abstract	Background and objectives: Despite the importance of provider continuity across healthcare settings, continuity among home care workers who provide hands-on long-term care is understudied. This project describes home care worker continuity, identifies factors associated with increased continuity, and examines associations between continuity and client outcomes. Research design and methods: We conducted a retrospective cohort study of clients receiving Medicaid-funded home-based long-term care ( Results: While home care worker continuity was lowest for clients receiving the most weekly care hours, a range of continuity existed across all levels of care need. Those who were male, older, Asian/Pacific Islander/Native American, cognitively impaired, and functionally impaired had lower continuity. Higher home care worker continuity was significantly associated ( Discussion and implications: The finding that home care worker continuity is associated with the health and well-being of home-based long-term care clients underscores the importance of building high-quality relationships in long-term care. Continued efforts are necessary to understand and advance home care worker continuity and to identify other aspects of the home care experience that benefit those receiving long-term care at home.
Language	English
Publishing date	2024-02-23
Publishing country	England
Document type	Journal Article
ISSN	2399-5300
ISSN (online)	2399-5300
DOI	10.1093/geroni/igae024
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Article ; Online: Intracapillary LPL levels in brown adipose tissue, visualized with an antibody-based approach, are regulated by ANGPTL4 at thermoneutral temperatures.

Song, Wenxin / Yang, Ye / Heizer, Patrick / Tu, Yiping / Weston, Thomas A / Kim, Joonyoung R / Munguia, Priscilla / Jung, Hyesoo / Fong, Jared L-C / Tran, Caitlyn / Ploug, Michael / Beigneux, Anne P / Young, Stephen G / Fong, Loren G

Proceedings of the National Academy of Sciences of the United States of America

2023 Volume 120, Issue 8, Page(s) e2219833120

Abstract: ... thermoneutral conditions (30 °C). To pursue this objective, we developed an antibody-based method to quantify ... in a direct fashion) LPL levels inside capillaries. At 30 °C, intracapillary LPL levels fell sharply ... of TRLs along capillaries. ANGPTL4 expression in BAT increased fourfold at 30 °C, suggesting a potential ...

Abstract	Lipoprotein lipase (LPL) is secreted into the interstitial spaces by parenchymal cells and then transported into capillaries by GPIHBP1. LPL carries out the lipolytic processing of triglyceride (TG)-rich lipoproteins (TRLs), but the tissue-specific regulation of LPL is incompletely understood. Plasma levels of TG hydrolase activity after heparin injection are often used to draw inferences about intravascular LPL levels, but the validity of these inferences is unclear. Moreover, plasma TG hydrolase activity levels are not helpful for understanding LPL regulation in specific tissues. Here, we sought to elucidate LPL regulation under thermoneutral conditions (30 °C). To pursue this objective, we developed an antibody-based method to quantify (in a direct fashion) LPL levels inside capillaries. At 30 °C, intracapillary LPL levels fell sharply in brown adipose tissue (BAT) but not heart. The reduced intracapillary LPL levels were accompanied by reduced margination of TRLs along capillaries. ANGPTL4 expression in BAT increased fourfold at 30 °C, suggesting a potential explanation for the lower intracapillary LPL levels. Consistent with that idea,
MeSH term(s)	Animals ; Mice ; Adipose Tissue/metabolism ; Adipose Tissue, Brown/metabolism ; Antibodies/metabolism ; Lipoprotein Lipase/metabolism ; Receptors, Lipoprotein/metabolism ; Temperature ; Triglycerides/metabolism
Chemical Substances	Antibodies ; Lipoprotein Lipase (EC 3.1.1.34) ; Receptors, Lipoprotein ; Triglycerides ; Angptl4 protein, mouse
Language	English
Publishing date	2023-02-14
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.2219833120
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