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1.
Heliyon ; 9(11): e21668, 2023 Nov.
Article in English | MEDLINE | ID: mdl-38027635

ABSTRACT

The use of the automated dispensing cabinet (ADC) for drug distribution in hospitals has become increasingly common and has numerous benefits. This retrospective study assessed the effectiveness of an ADC that uses integrated information technology in the drug distribution process in a surgical unit as part of a smart medical process improvement project at Taichung Veterans General Hospital in 2019. The outcomes include medicine delivery time, working time of healthcare professionals, transportation manpower, dispensing errors, and satisfaction of nursing staff with the medication distribution process. After ADC implementation, the average waiting time of standing orders decreased significantly for both on and off duty periods (40.0 ± 27.6 to 3.0 ± 3.9 min, P < 0.001; 45.2 ± 25.8 to 2.9 ± 2.9 min, P < 0.001; respectively). Similar results were observed with immediate or temporary medication orders (54.4 ± 31.5 to 2.0 ± 3.0 min, P < 0.001; 64.0 ± 47.5 to 1.5 ± 1.8 min, P < 0.001; respectively). The average time spent by operation room and post-operation room (OR/POR) nurses on communicating with ward nurses for medication delivery to OR/POR was shortened by 46.9 ± 4.4 h per month, and the average time pharmacists spent on dispensing immediate or temporary medication orders was shortened by 5.6 ± 0.2 h per month. The satisfaction of nursing staff with the OR/POR drug delivery process was significantly improved after ADC implementation (3.2 ± 0.8 vs 4.2 ± 0.7, P < 0.001). Our results showed that ADC implementation in surgical units simplified drug delivery processes, shortened drug delivery time, improved drug delivery timeliness for surgical patients, decreased dispensing errors, and increased nursing staff satisfaction. In conclusion, the implementation of ADC was beneficial for surgical units. To the best of our knowledge, there have been no studies on a similar ADC system.

2.
Biosensors (Basel) ; 13(10)2023 Oct 16.
Article in English | MEDLINE | ID: mdl-37887122

ABSTRACT

An effective early diagnosis is important for rheumatoid arthritis (RA) management. This study reveals a novel RA detection method using bacteriorhodopsin as a photoelectric transducer, a light-driven proton pump in purple membranes (PMs). It was devised by covalently conjugating a PM monolayer-coated electrode with a citrullinated-inter-alpha-trypsin inhibitor heavy chain 3 (ITIH3)542-556 peptide that recognizes the serum RA-associated autoantibodies. The direct serum coating decreased the photocurrents in the biosensor, with the reduction in the photocurrent caused by coating with an RA-patient serum that is significantly larger than that with a healthy-control serum (38.1% vs. 20.2%). The difference in the reduction in the photocurrent between those two serum groups widened after the serum-coated biosensor was further labeled with gold nanoparticle (AuNP)-conjugated anti-IgA (anti-IgA-AuNP) (53.6% vs. 30.6%). Both atomic force microscopic (AFM) and Raman analyses confirmed the sequential peptide, serum, and anti-IgA-AuNP coatings on the PM-coated substrates. The reductions in the photocurrent measured in both the serum and anti-IgA-AuNPs coating steps correlated well with the results using commercial enzyme-linked immunosorbent assay kits (Spearman rho = 0.805 and 0.787, respectively), with both a sensitivity and specificity close to 100% in both steps. It was shown that an RA diagnosis can be performed in either a single- or two-step mode using the developed biosensor.


Subject(s)
Arthritis, Rheumatoid , Bacteriorhodopsins , Biosensing Techniques , Metal Nanoparticles , Humans , Gold , Arthritis, Rheumatoid/diagnosis , Peptides , Enzyme-Linked Immunosorbent Assay
3.
Sensors (Basel) ; 23(17)2023 Aug 27.
Article in English | MEDLINE | ID: mdl-37687909

ABSTRACT

A uniformly oriented purple membrane (PM) monolayer containing photoactive bacteriorhodopsin has recently been applied as a sensitive photoelectric transducer to assay color proteins and microbes quantitatively. This study extends its application to detecting small molecules, using adenosine triphosphate (ATP) as an example. A reverse detection method is used, which employs AuNPs labeling and specific DNA strand displacement. A PM monolayer-coated electrode is first covalently conjugated with an ATP-specific nucleic acid aptamer and then hybridized with another gold nanoparticle-labeled nucleic acid strand with a sequence that is partially complementary to the ATP aptamer, in order to significantly minimize the photocurrent that is generated by the PM. The resulting ATP-sensing chip restores its photocurrent production in the presence of ATP, and the photocurrent recovers more effectively as the ATP concentration increases. Direct and single-step ATP detection is achieved in 15 min, with detection limits of 5 nM and a dynamic range of 5 nM-0.1 mM. The sensing chip exhibits high selectivity against other ATP analogs and is satisfactorily stable in storage. The ATP-sensing chip is used to assay bacterial populations and achieves a detection limit for Bacillus subtilis and Escherichia coli of 102 and 103 CFU/mL, respectively. The demonstration shows that a variety of small molecules can be simultaneously quantified using PM-based biosensors.


Subject(s)
Bacteriorhodopsins , Metal Nanoparticles , Nucleic Acids , Gold , DNA , Adenosine Triphosphate , Escherichia coli
4.
Mol Neurobiol ; 58(5): 2396-2406, 2021 May.
Article in English | MEDLINE | ID: mdl-33421017

ABSTRACT

Proper development of neuronal cells is important for brain functions, and impairment of neuronal development may lead to neuronal disorders, implying that improvement in neuronal development may be a therapeutic direction for these diseases. Huntington's disease (HD) is a neurodegenerative disease characterized by impairment of neuronal structures, ultimately leading to neuronal death and dysfunctions of the central nervous system. Based on previous studies, fibroblast growth factor 9 (FGF9) may provide neuroprotective functions in HD, and FGFs may enhance neuronal development and neurite outgrowth. However, whether FGF9 can provide neuronal protective functions through improvement of neuronal morphology in HD is still unclear. Here, we study the effects of FGF9 on neuronal length in HD and attempt to understand the related working mechanisms. Taking advantage of striatal cell lines from HD knock-in mice, we found that FGF9 increases total neuronal length and upregulates several structural and synaptic proteins under HD conditions. In addition, activation of nuclear factor kappa B (NF-kB) signaling by FGF9 was observed to be significant in HD cells, and blockage of NF-kB leads to suppression of these structural and synaptic proteins induced by FGF9, suggesting the involvement of NF-kB signaling in these effects of FGF9. Taken these results together, FGF9 may enhance total neuronal length through upregulation of NF-kB signaling, and this mechanism could serve as an important mechanism for neuroprotective functions of FGF9 in HD.


Subject(s)
Corpus Striatum/drug effects , Fibroblast Growth Factor 9/pharmacology , Huntington Disease/metabolism , NF-kappa B/metabolism , Neuronal Outgrowth/drug effects , Neurons/drug effects , Animals , Cell Line , Corpus Striatum/metabolism , Disease Models, Animal , Mice , Neurons/metabolism , Signal Transduction/drug effects , Up-Regulation/drug effects
5.
Life Sci ; 267: 118952, 2021 Feb 15.
Article in English | MEDLINE | ID: mdl-33383048

ABSTRACT

AIMS: Huntington's disease (HD) is a neurodegenerative disease that causes deficits in neurite outgrowth, which suggests that enhancement of neurite outgrowth is a potential direction by which to improve HD. Our previous publications showed that fibroblast growth factor 9 (FGF9) provides anti-apoptosis and anti-oxidative functions in striatal cell models of HD through the extracellular signal-regulated kinases (ERK) pathway, and FGF9 also stimulates cytoskeletons to enhance neurite outgrowth via nuclear factor kappa B (NF-kB) signaling. In this study, we further demonstrate the importance of the ERK pathway for the neurite outgrowth induced by FGF9 in HD striatal models. MATERIALS AND METHODS: FGF9 was treated with ERK (U0126) or NF-kB (BAY11-7082) inhibitors in STHdhQ7/Q7 and STHdhQ111/Q111 striatal knock-in cell lines to examine neurite outgrowth, cytoskeletal markers, and synaptic proteins via immunofluorescence staining and Western blotting. NF-kB activity was analyzed by NF-kB promoter reporter assay. KEY FINDINGS: Here, we show that suppression of ERK signaling significantly inhibits FGF9-induced neurite outgrowth, cytoskeletal markers, and synaptic proteins in HD striatal cells. In addition, we also show suppression of ERK signaling significantly decreases FGF9-induced NF-kB activation, whereas suppression of NF-kB does not decrease FGF9-induced ERK signaling. These results suggest that FGF9 activates ERK signaling first, stimulates NF-kB upregulation, and then enhances neurite outgrowth in HD striatal cells. SIGNIFICANCE: We elucidate the more detailed mechanisms of neurite outgrowth enhanced by FGF9 in these HD striatal cells. This study may provide insights into targeting neurite outgrowth for HD therapy.


Subject(s)
Fibroblast Growth Factor 9/metabolism , Fibroblast Growth Factor 9/pharmacology , MAP Kinase Signaling System/drug effects , Neurites/metabolism , Animals , Butadienes/pharmacology , Cell Line , Cells, Cultured , Corpus Striatum/metabolism , Enzyme Inhibitors/pharmacology , Fibroblast Growth Factor 9/antagonists & inhibitors , Humans , Huntingtin Protein/genetics , Huntingtin Protein/metabolism , Huntington Disease/metabolism , Mice , Mice, Transgenic , NF-kappa B/antagonists & inhibitors , NF-kappa B/metabolism , Neurites/drug effects , Neuronal Outgrowth/physiology , Nitriles/pharmacology , Nuclear Proteins/metabolism , Oxidative Stress/drug effects , Recombinant Proteins/pharmacology , Signal Transduction , Sulfones/pharmacology
6.
Sensors (Basel) ; 20(24)2020 Dec 18.
Article in English | MEDLINE | ID: mdl-33353006

ABSTRACT

Glycated hemoglobin (HbA1c) levels are an important index for the diagnosis and long-term control of diabetes. This study is the first to use a direct and label-free photoelectric biosensor to determine HbA1c using bacteriorhodopsin-embedded purple membranes (PM) as a transducer. A biotinylated PM (b-PM) coated electrode that is layered with protein A-oriented antibodies against hemoglobin (Hb) readily captures non-glycated Hb (HbA0) and generates less photocurrent. The spectra of bacteriorhodopsin and Hb overlap so the photocurrent is reduced because of the partial absorption of the incident light by the captured Hb molecules. Two HbA0 and HbA1c aptasensors that are prepared by conjugating specific aptamers on b-PM coated electrodes single-step detect HbA0 and HbA1c in 15 min, without cross reactivity, with detection limits of ≤0.1 µg/mL and a dynamic range of 0.1-100 µg/mL. Both aptasensors exhibit high selectivity and long-term stability. For the clinical samples, HbA0 concentrations and HbA1c levels that are measured with aptasensors correlate well with total Hb concentrations and the HbA1c levels that are determined using standard methods (correlation gradient = 0.915 ± 0.004 and 0.981 ± 0.001, respectively). The use of these aptasensors for diabetes care is demonstrated.


Subject(s)
Bacteriorhodopsins , Biosensing Techniques , Animals , Glycated Hemoglobin/analysis , Humans , Rabbits , Transducers
7.
Free Radic Biol Med ; 130: 256-266, 2019 01.
Article in English | MEDLINE | ID: mdl-30391672

ABSTRACT

Huntington's disease (HD) is a heritable neurodegenerative disorder, and has been characterized as an increase of oxidative stress in brain regions. In our previous results, we showed fibroblast growth factor 9 (FGF9) provides neuroprotective functions to suppress cell death in HD striatal cells dominantly through ERK signalling. However, whether the working mechanism of FGF9 is related to anti-oxidative stress in HD is still unknown. In this study, STHdhQ7/Q7 (Q7) and STHdhQ111/Q111 (Q111) striatal knock-in cell lines were used to examine the neuroprotective effects of FGF9 against oxidative stress in HD. Results show that FGF9 alleviates oxidative stress induced by starvation in Q7 and Q111 cells. The treatment of FGF9 not only induces upregulation and activation of nuclear factor erythroid 2-like 2 (Nrf2), a critical transcription factor for anti-oxidative stress, but also further upregulates its downstream targets, such as superoxide dismutase 2, gamma-glutamylcysteine synthetase and glutathione reductase. Furthermore, blockage of the Nrf2 pathway abolishes the anti-oxidative functions of FGF9, and inhibition of ERK signalling reduces the activation of the FGF9-Nrf2 pathway, resulting in higher level of oxidative stress in HD cells. These results support the neuroprotective effects of FGF9 against oxidative stress through the ERK-Nrf2 pathway, and imply one of potential strategies for therapy of HD.


Subject(s)
Antioxidants/pharmacology , Brain/metabolism , Fibroblast Growth Factor 9/genetics , Huntington Disease/drug therapy , NF-E2-Related Factor 2/genetics , Animals , Brain/pathology , Cell Line , Gene Expression Regulation/drug effects , Glutamate-Cysteine Ligase/genetics , Glutathione Reductase/genetics , Humans , Huntingtin Protein/genetics , Huntington Disease/genetics , Huntington Disease/pathology , MAP Kinase Signaling System/genetics , Mice , Neuroprotective Agents/pharmacology , Oxidative Stress/drug effects , Superoxide Dismutase/genetics , Visual Cortex/metabolism , Visual Cortex/pathology
8.
Sensors (Basel) ; 18(12)2018 Dec 18.
Article in English | MEDLINE | ID: mdl-30567418

ABSTRACT

Bacteriorhodopsin-embedded purple membranes (PM) have been demonstrated to be a sensitive photoelectric transducer for microbial detection. To efficiently prepare versatile BR-based immunosensors with protein A as antibody captures, a large, high-coverage, and uniformly oriented PM monolayer was fabricated on an electrode as an effective foundation for protein A conjugation through bis-NHS esters, by first affinity-coating biotinylated PM on an aminated surface using a complex of oxidized avidin and graphene oxide as the planar linker and then washing the coating with a shear flow. Three different polyclonal antibodies, each against Escherichia coli, Lactobacillus acidophilus, and Streptococcus mutans, respectively, were individually, effectively and readily adsorbed on the protein A coated electrodes, leading to selective and sensitive quantitative detection of their respective target cells in a single step without any labeling. A single-cell detection limit was achieved for the former two cells. AFM, photocurrent, and Raman analyses all displayed each fabricated layer as well as the captured bacteria, with AFM particularly revealing the formation of a massive continuous PM monolayer on aminated mica. The facile cell-membrane monolayer fabrication and membrane surface conjugation techniques disclosed in this study may be widely applied to the preparation of different biomembrane-based biosensors.

9.
Cell Physiol Biochem ; 48(2): 605-617, 2018.
Article in English | MEDLINE | ID: mdl-30021209

ABSTRACT

BACKGROUND/AIMS: Huntington's disease (HD) is a heritable neurodegenerative disorder, and there is no cure for HD to date. A type of fibroblast growth factor (FGF), FGF9, has been reported to play prosurvival roles in other neurodegenerative diseases, such as Parkinson's disease and Alzheimer's disease. However, the effects of FGF9 on HD is still unknown. With many similarities in the cellular and pathological mechanisms that eventually cause cell death in neurodegenerative diseases, we hypothesize that FGF9 might provide neuroprotective functions in HD. METHODS: In this study, STHdhQ7/Q7 (WT) and STHdhQ111/Q111 (HD) striatal knock-in cell lines were used to evaluate the neuroprotective effects of FGF9. Cell proliferation, cell death and neuroprotective markers were determined via the MTT assay, propidium iodide staining and Western blotting, respectively. The signaling pathways regulated by FGF9 were demonstrated using Western blotting. Additionally, HD transgenic mouse models were used to further confirm the neuroprotective effects of FGF9 via ELISA, Western blotting and immunostaining. RESULTS: Results show that FGF9 not only enhances cell proliferation, but also alleviates cell death as cells under starvation stress. In addition, FGF9 significantly upregulates glial cell line-derived neurotrophic factor (GDNF) and an anti-apoptotic marker, Bcl-xL, and decreases the expression level of an apoptotic marker, cleaved caspase 3. Furthermore, FGF9 functions through ERK, AKT and JNK pathways. Especially, ERK pathway plays a critical role to influence the effects of FGF9 toward cell survival and GDNF production. CONCLUSIONS: These results not only show the neuroprotective effects of FGF9, but also clarify the critical mechanisms in HD cells, further providing an insight for the therapeutic potential of FGF9 in HD.


Subject(s)
Extracellular Signal-Regulated MAP Kinases/metabolism , Fibroblast Growth Factor 9/pharmacology , MAP Kinase Signaling System/drug effects , Animals , Apoptosis/drug effects , Butadienes/pharmacology , Caspase 3/metabolism , Cell Line , Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors , Glial Cell Line-Derived Neurotrophic Factor/metabolism , Huntington Disease/metabolism , Huntington Disease/pathology , JNK Mitogen-Activated Protein Kinases/metabolism , Mice , Mice, Transgenic , Nitriles/pharmacology , Proto-Oncogene Proteins c-akt/metabolism , Visual Cortex/cytology , Visual Cortex/drug effects , Visual Cortex/metabolism , bcl-X Protein/metabolism
11.
Biosens Bioelectron ; 91: 24-31, 2017 May 15.
Article in English | MEDLINE | ID: mdl-27987407

ABSTRACT

A photoelectric immunosensor using purple membranes (PM) as the transducer, which contains photoactive bacteriorhodopsin, is here first demonstrated for direct and label-free microbial detection. Biotinylated polyclonal antibodies against Escherichia coli were immobilized on a PM-coated electrode through further surface biotinylation and bridging avidin or NeutrAvidin. The photocurrent generated by the antibody-coated sensor was reduced after incubation with E. coli K-12 cultures, with the reduction level increased with the culture populations. The immunosensor prepared via NeutrAvidin exhibited much better selectivity than the one prepared via avidin, recognizing almost none of the tested Gram-positive bacteria. Cultures with populations ranging from 1 to 107CFU/10mL were detected in a single step without any preprocessing. Both AFM and Raman analysis confirmed the layer-by-layer fabrication of the antibody-coated substrates as well as the binding of microorganisms. By investigating the effect of illumination orientation and simulating the photocurrent responses with an equivalent circuit model containing a chemical capacitance, we suggest that the photocurrent reduction was primarily caused by the light-shielding effect of the captured bacteria. Using the current fabrication technique, versatile bacteriorhodopsin-based photoelectric immunosensors can be readily prepared to detect a wide variety of biological cells.


Subject(s)
Antibodies, Immobilized/chemistry , Bacteria/isolation & purification , Bacteriorhodopsins/chemistry , Biosensing Techniques/methods , Halobacterium salinarum/chemistry , Purple Membrane/chemistry , Bacterial Infections/microbiology , Biosensing Techniques/instrumentation , Electrodes , Equipment Design , Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Humans
12.
Oncotarget ; 6(17): 14940-52, 2015 Jun 20.
Article in English | MEDLINE | ID: mdl-25909285

ABSTRACT

Foretinib, a multiple kinase inhibitor undergoing clinical trials, could suppress the activity of hepatocyte growth factor (HGF) receptor c-MET and vascular endothelial growth factor receptor-2 (VEGFR-2). In addition, Foretinib may inhibit two critical lymphangiogenic signaling receptors VEGFR-3 and TIE-2. However, the effect of Foretinib on lymphatic endothelial cells (LECs) in vitro and lymphangiogenesis in vivo is still unknown. We found Foretinib decreased basal- and HGF-induced c-MET activity at low concentrations. However, Foretinib only reduced the proliferation of pancreatic cancer cells at high concentration reflecting the intrinsic chemoresistance of pancreatic cancer cells. Foretinib inhibited VEGF-A, VEGF-C and Angiopoetin-2 (ANG-2)-stimulated tube formation and sprouting of LECs by reducing VEGFR-2, VEGFR-3 and TIE-2 activation and increased apoptosis of LECs. In xenograft animal study, Foretinib suppressed tumor growth by inhibiting proliferation, angiogenesis and lymphangiogenesis. Additionally, Foretinib inhibited angiogenesis and lymphangiogenesis more significantly and exhibited low detrimental effect in orthotopic animal study. Collectively, we suggested that Foretinib simultaneously inhibits cancer cells and LECs to reduce pancreatic tumor growth in vivo and demonstrated for the first time that Foretinib suppresses angiogenesis and lymphangiogenesis by blocking VEGFR-2/3 and TIE-2 signaling.


Subject(s)
Anilides/pharmacology , Neovascularization, Pathologic/prevention & control , Pancreatic Neoplasms/drug therapy , Quinolines/pharmacology , Receptor, TIE-2/metabolism , Vascular Endothelial Growth Factor Receptor-2/metabolism , Vascular Endothelial Growth Factor Receptor-3/metabolism , Angiopoietin-2/pharmacology , Animals , Blotting, Western , Cell Line, Tumor , Cells, Cultured , Extracellular Signal-Regulated MAP Kinases/metabolism , Humans , Lymphangiogenesis/drug effects , Mice, Inbred NOD , Mice, SCID , Neovascularization, Pathologic/metabolism , Neovascularization, Physiologic/drug effects , Pancreatic Neoplasms/blood supply , Pancreatic Neoplasms/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins c-met/metabolism , Signal Transduction/drug effects , Tumor Burden/drug effects , Vascular Endothelial Growth Factor A/pharmacology , Vascular Endothelial Growth Factor C/pharmacology , Xenograft Model Antitumor Assays
13.
Colloids Surf B Biointerfaces ; 116: 482-8, 2014 Apr 01.
Article in English | MEDLINE | ID: mdl-24561502

ABSTRACT

The effect of graphene oxide (GO) on the surface fabrication of purple membranes (PM) containing photosensitive bacteriorhodopsin is first reported in this study. GO was initially modified with biotin and then coupled with oxidized avidin to generate a GO-avidin complex, which was subsequently used as a linker to immobilize biotinylated PM (b-PM) onto amine-functionalized supports. Indium-tin-oxide glass coated with the GO-avidin complex was more hydrophilic than the electrode coated only with oxidized avidin, and the successive b-PM adsorption yielded a 1.4-fold higher (410 nA/cm(2)) photoelectric activity. AFM analysis on mica revealed that the GO-avidin complex layer had less surface roughness and dissipation energy than the pure oxidized avidin linker layer. For subsequent b-PM fabrication, GO addition not only reduced the stacking of immobilized b-PM patches but also improved their interior compactness and surface smoothness. This study demonstrates a convenient way to introduce GO into PM fabrication technology to provide enhanced surface morphology and photoelectric activity.


Subject(s)
Bacteriorhodopsins/chemistry , Graphite/chemistry , Oxides/chemistry , Purple Membrane/chemistry , Halobacterium salinarum/chemistry , Molecular Structure , Surface Properties
14.
Protein Expr Purif ; 89(2): 219-24, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23583309

ABSTRACT

Purple membrane (PM) is a part of cytoplasmic membrane in certain extreme halophilic microorganisms belonging to Domain Archaea. It transduces light energy to generate proton gradient for ATP synthesis in the microorganisms. Bacteriorhodopsin (BR) is the only protein in PM responsible for the generation of proton gradient. Generally, PM was purified from Halobacterium salinarum via a tedious and lengthy sucrose density gradient ultracentrifugation (SGU). In this work, a facile method based on polyethyleneglycol (PEG)-phosphate aqueous-two- phase extraction system (ATPS) was employed to purify PM from cell lysate of H. salinarum. The results showed that PM could be completely recovered from the interface of PEG-phosphate ATPS with BR purity ca 94.1% as measured by UV-visible absorption spectra. In comparison with PM obtained by SGU, the PM isolated by ATPS could achieve the same level of purity and photocurrent activity (ca 177.2nA/µgBR/cm(2)) as analyzed by SDS-PAGE and photocurrent measurement, respectively. The easily scalable and straightforward ATPS procedure demonstrated that PM can be purified and recovered more cost-effectively with a significantly reduced operation time that should lead to broader range applications of PM possible.


Subject(s)
Archaeal Proteins/isolation & purification , Archaeal Proteins/metabolism , Bacteriorhodopsins/isolation & purification , Bacteriorhodopsins/metabolism , Halobacterium salinarum/metabolism , Purple Membrane/metabolism , Cell Fractionation , Electrophoresis, Polyacrylamide Gel , Equipment Design , Halobacterium salinarum/chemistry , Phosphates/chemistry , Photochemistry/instrumentation , Polyethylene Glycols/chemistry , Purple Membrane/chemistry , Ultracentrifugation
15.
J Clin Endocrinol Metab ; 98(5): E837-46, 2013 May.
Article in English | MEDLINE | ID: mdl-23482612

ABSTRACT

CONTEXT: Uterine leiomyoma, the most common tumors found in the women of the reproductive age, may cause abnormal uterine bleeding and be life threatening. Compared with myometrium, leiomyoma contains excessive extracellular matrix (ECM). However, the pathological roles of ECM in the development of leiomyoma remain largely unknown. Integrins are the major adhesion molecules on cell surface to interact with ECM. The interactions of ECM with integrins regulate cell adhesion and initiate signals for cell growth, differentiation, and migration. OBJECTIVE: The aim of this study was to investigate the expression and functional role of integrin-ß1 in leiomyoma pathogenesis. DESIGN: Levels of integrin-ß1 protein were determined by Western blotting in paired normal and leiomyomal tissues (n = 15). Knockdown of integrin-ß1 and inhibition of ECM-integrin interaction by disintegrin were used to evaluate the impact of integrin-ß1 in cell adhesion, spreading, and proliferation. RESULTS: Levels of integrin-ß1 were significantly up-regulated in leiomyomal cells compared with their normal counterparts. Knockdown of integrin-ß1 did not affect cell adhesion on fibronectin or laminin matrix but significantly inhibits cell spreading ability. Consistent with this notion, the phosphorylation of focal adhesion kinase and the recruitment of paxillin to the focal contact were decreased in integrin-ß1 knockdown cells, which attenuates contraction force. The inability of cell spreading leads to inhibition of cyclin D1 expression and impedes cell cycle progression. More importantly, disruption of ECM-integrin interaction by the small protein, disintegrin inhibited cyclin D1 expression and cell proliferation. CONCLUSION: These data demonstrate that integrin-ß1 is a critical ligand to enhance cell-ECM contact force and thus promotes cell proliferation. Disruption of ECM-integrin-ß1 signaling may serve as an option to inhibit the progression of leiomyoma.


Subject(s)
Antigens, Neoplasm/metabolism , Integrin beta1/metabolism , Leiomyoma/metabolism , Up-Regulation , Uterine Neoplasms/metabolism , Antigens, Neoplasm/chemistry , Antigens, Neoplasm/genetics , Cell Adhesion , Cell Movement , Cell Proliferation , Cell Shape , Cyclin D1/metabolism , Disease Progression , Disintegrins/metabolism , Extracellular Matrix Proteins/metabolism , Female , Focal Adhesion Protein-Tyrosine Kinases/metabolism , Humans , Integrin beta1/chemistry , Integrin beta1/genetics , Leiomyoma/pathology , Leiomyoma/physiopathology , Leiomyoma/surgery , Paxillin/metabolism , Phosphorylation , Protein Processing, Post-Translational , RNA Interference , RNA, Small Interfering , Tumor Cells, Cultured , Uterine Neoplasms/pathology , Uterine Neoplasms/physiopathology , Uterine Neoplasms/surgery
16.
Nurse Educ Today ; 31(1): 18-23, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20406717

ABSTRACT

The purpose of this study was to identify the relationship between learning styles and age among nursing students in a two-year, a five-year associate degree of nursing (ADN) program, and a two-year bachelor of science in nursing (BSN) program in Taiwan. The Chinese version of the Myers-Briggs Type Indicator (MBTI) Form M was used to measure individual preferences in four dichotomous dimensions of Jungian theory: extraversion/introversion, sensing/intuition, thinking/feeling, and judging/perceiving. The study sample included 331 nursing students. The analysis of the data revealed that the most common learning styles were introversion, sensing, thinking, and judging (ISTJ) and introversion, sensing, feeling, and judging (ISFJ). The findings indicated that the SJs comprised 43.0% of the participating nursing students. SJs are highly preferred in the field of nursing. However, the ages of nursing students were not significantly related to their learning styles. The findings suggested that the participating nursing students were homogeneous. We recommend the use of a large sample for further studies. The awareness and understanding of individual differences is of great importance in tailoring each learning style to benefit educators and learners, thereby enhancing nursing education.


Subject(s)
Education, Nursing/methods , Learning , Schools, Nursing , Students, Nursing/psychology , Teaching/methods , Adolescent , Adult , Age Factors , Curriculum , Education, Nursing, Associate , Education, Nursing, Baccalaureate , Educational Measurement , Educational Status , Humans , Jungian Theory , Male , Models, Educational , Nursing Theory , Surveys and Questionnaires , Taiwan , Young Adult
17.
Anal Biochem ; 392(1): 1-7, 2009 Sep 01.
Article in English | MEDLINE | ID: mdl-19464992

ABSTRACT

A biotin-coated quartz crystal microbalance (QCM) chip was prepared by dip-coating a long-chain alkanethiol-modified crystal with precoupled dextran-biotin hydrogels. The resulting biotin chip was used to affinity-immobilize streptavidin (SAv) and was then further employed for various biosensor assays. First, the SAv chip allowed efficient on-line binding of biotinylated bovine serum albumin (bBSA), followed by a sensitive and specific response toward anti-bovine serum albumin (BSA) antibodies. Three consecutive immunoassays were reproducibly demonstrated with a single chip. The apparent binding kinetics with k(on)=5.9 microM(-1) h(-1), k(off)=10.1 h(-1), and K(D)=1.71 microM was readily resolved by fitting the real-time sensorgrams. Second, the capability of the SAv chip to selectively recognize recombinant Escherichia coli with flagella displaying an artificial SAv binding peptide, Strep-tag II, was demonstrated by QCM analysis and verified by scanning transmission electron microscope (STEM) image analysis with biotin-coated gold nanoparticles as the label. Finally, the affinity of the cell-displayed Strep-tag II peptide to surface-coated SAv, K(D)=6.8 x 10(8) CFU/ml, was resolved on-line using equilibrium binding kinetics by QCM. This study presents an easy, economical, and reliable method of preparing high-performance SAv-coated biotin chips with potential for application in real-time repetitive immunoassays, on-line binding kinetics studies, and high-affinity peptide screening.


Subject(s)
Biosensing Techniques/methods , Biotin/analysis , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Immunoassay/methods , Peptide Library , Quartz/chemistry , Animals , Biosensing Techniques/instrumentation , Biotin/chemistry , Cattle , Crystallization , Escherichia coli/chemistry , Immunoassay/instrumentation , Kinetics , Microchip Analytical Procedures/methods , Microscopy, Electron, Transmission , Molecular Structure , Reproducibility of Results , Serum Albumin, Bovine/immunology
18.
Cancer Nurs ; 31(5): 363-9, 2008.
Article in English | MEDLINE | ID: mdl-18772661

ABSTRACT

Oral mucositis (OM) is a frequent and potentially severe complication of cancer chemotherapy. The aim of this descriptive, cross-sectional study was to better understand patients' experiences and perceptions of chemotherapy-induced OM. Fifty-seven adult patients in a chemotherapy day unit who had completed at least 2 cycles of chemotherapy within the previous 12 months and who were receiving chemotherapy at the time of study completed the questionnaire (response rate, 86.3%). Results demonstrated that 75.4% of participants (N = 57) had experienced at least one episode of OM since their first chemotherapy. Dry lips were the most common symptom of OM (n = 20, 54.1% previously; n = 14, 73.7% currently). Ulcerated mucosa was regarded as the most significant problem caused by OM (n = 14, 87.5% previously; n = 8, 100% currently), whereas pain on swallowing was considered as the most distressing mucositis-related effect (n = 14, 87.5% previously; n = 4, 100% currently). The findings revealed a high incidence of chemotherapy-induced OM. Chemotherapy-related mouth problems were perceived as problematic and caused varying degrees of distress. A systematic oral assessment method can be used in future studies with a cohort of patients to further generalize the pattern of patients' experiences of OM.


Subject(s)
Antineoplastic Agents/adverse effects , Neoplasms/drug therapy , Patient Satisfaction , Quality of Life , Stomatitis/chemically induced , Adaptation, Psychological , Adult , Aged , Aged, 80 and over , Antineoplastic Agents/therapeutic use , Cross-Sectional Studies , Diagnosis, Oral , Female , Humans , Male , Middle Aged , Neoplasms/complications , Oncology Service, Hospital , Pilot Projects , Stomatitis/psychology , Surveys and Questionnaires
19.
Biotechnol Prog ; 23(3): 741-8, 2007.
Article in English | MEDLINE | ID: mdl-17469846

ABSTRACT

Two different hydrogel-coated streptavidin (SAv) piezoelectric chips were investigated. One was directly prepared by immobilizing SAv molecules covalently onto a dextran-modified crystal, and the other one was indirectly prepared by physically adsorbing SAv onto a biotin-linked dextran surface. The covalent preparation yielded 80% more SAv-binding and better subsequent adsorption of biotinylated bovine serum albumin (bBSA). Both chips displayed the best binding affinity with bBSA at pH 5.0 in a flow injection analysis and exhibited reproductive real-time response during layer-by-layer assembly of a bBSA and SAv multilayer film. In the multilayer assembly, approximately 3-7 SAv molecules were captured by each immobilized bBSA, and the estimated apparent KD values of the binding of flowing bBSA with surface SAv were 0.24 and 0.11 microM in the first two cycles of the covalently prepared chip, respectively. Two Escherichia coli cells, each flagellum-displaying Strep-tag I and Strep-tag II, respectively, were selectively detected by both kinds of SAv chips. These studies suggest the potential application of both chips in real-time screening SAv affinity ligands from a cell-display random peptide library.


Subject(s)
Biosensing Techniques/methods , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Streptavidin/chemistry , Animals , Biotinylation , Cattle , Dextrans/chemistry , Escherichia coli/chemistry , Escherichia coli/cytology , Escherichia coli/metabolism , Hydrogen-Ion Concentration , Models, Chemical , Protein Binding , Serum Albumin, Bovine/metabolism
20.
J Clin Endocrinol Metab ; 90(6): 3715-23, 2005 Jun.
Article in English | MEDLINE | ID: mdl-15769981

ABSTRACT

Pituitary tumor-transforming gene-1 (PTTG-1) is a novel protooncogene overexpressed in numerous cancer cell lines and cancers. In this study we elucidate the expression of PTTG-1 in uterine leiomyomas and its functional role in the development of this disease. By comparing 23 pairs of leiomyomas and matched pairs of myometria, we found that the expression of PTTG-1 is significantly elevated in leiomyoma. The expression of PTTG-1 is independent of the menstrual cycle and is not affected by ovarian hormones. In contrast, basic fibroblast growth factor (bFGF) time- and dose-dependently stimulates PTTG-1 expression, which results in increasing cell proliferation. Forced expression of PTTG-1 by transient transfection stimulates bFGF and VEGF expression as well as changes the expression pattern of cell cycle proteins. Western blotting analysis demonstrates that the expressions of PTTG-1, bFGF, and the cell proliferation marker, proliferating cell nuclear antigen, are positively correlated with each other, which supports the hypothesis that the positive feedback loop between PTTG-1 and bFGF increases leiomyoma cell proliferation. In summary, we have shown for the first time that PTTG-1 is up-regulated in human uterine leiomyomas and that the positive feedback loop between PTTG-1 and bFGF may be pivotal in the growth of leiomyoma cells.


Subject(s)
Gene Expression Regulation, Neoplastic , Leiomyoma/genetics , Neoplasm Proteins/genetics , Uterine Neoplasms/genetics , Base Sequence , DNA Primers , Feedback , Female , Fibroblast Growth Factor 2/genetics , Humans , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , Securin , Transcription, Genetic , Tumor Cells, Cultured
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