Nucleic acid analysis by sandwich hybridization

Nicholls, Peter J.; Malcolm, Alan D. B.

doi:10.1002/jcla.1860030210

Cited by 38 publications

(24 citation statements)

References 108 publications

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“…The present study shows that UCP-1, a member of mitochondrial transporter superfamily that increases energy expenditure by proton flux across the inner mitochondrial membrane (29), is up-regulated in interscapular BAT after L-NAME treatment. Because ␤-adrenergic stimulation of BAT has been shown to induce UCP-1 expression (30), the decreased adiposity after L-NAME treatment is likely due in part to increasing energy expenditure via induction of UCP-1 resulting from the enhanced sympathetic nerve activity.…”

Section: Discussionmentioning

confidence: 90%

Chronic Blockade of Nitric Oxide Synthesis Reduces Adiposity and Improves Insulin Resistance in High Fat-Induced Obese Mice

et al. 2007

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Genetic deletion of inducible nitric oxide synthase (NOS) in mice has been shown to improve high-fat diet (HFD)-induced insulin resistance. However, a pathophysiological role of endogenous nitric oxide (NO) in obesity-related insulin resistance remains controversial. To address this issue, we examined the metabolic phenotypes in HFD-induced obese mice with chronic blockade of NO synthesis by a NOS inhibitor, N(G)-nitro-l-arginine methyl ester (L-NAME). Six-week-old male C57BL/6j mice were provided free access to either a standard diet (SD) or a HFD and tap water with or without L-NAME (100 mg/kg.d) for 12 wk. L-NAME treatment significantly attenuated body weight gain of mice fed either SD or HFD without affecting calorie intake. L-NAME treatment in HFD-fed mice improved glucose tolerance and insulin sensitivity. HFD feeding induced inducible NOS mRNA expression, but not the other two NOS isoforms, in white adipose tissue (WAT) and skeletal muscle. L-NAME treatment up-regulated uncoupling protein-1 in brown adipose tissue of HFD-fed mice but down-regulated monocyte chemoattractant protein-1 and CD68 mRNAs levels in WAT. HFD feeding up-regulated leptin mRNA levels but conversely down-regulated adiponectin mRNA levels in WAT, but these effects were unaffected by L-NAME treatment. Moreover, L-NAME treatment also increased peroxisome proliferator-uncoupling protein-3 mRNA levels in skeletal muscles of HFD-fed mice. Increased urinary excretion of norepinephrine after HFD feeding was augmented in L-NAME-treated mice. Insulin-stimulated tyrosine phosphorylation of insulin receptor substrate-1 and serine phosphorylation of Akt/Akt2 in soleus muscle was markedly impaired in HFD-fed mice but reversed by L-NAME treatment. In conclusion, chronic NOS blockade by L-NAME in mice ameliorates HFD-induced adiposity and glucose intolerance, accompanied by reduced adipose inflammation and improved insulin signaling in skeletal muscle, suggesting that endogenous NO plays a modulatory role in the development of obesity-related insulin resistance.

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Section: Discussionmentioning

confidence: 90%

Chronic Blockade of Nitric Oxide Synthesis Reduces Adiposity and Improves Insulin Resistance in High Fat-Induced Obese Mice

et al. 2007

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“…There are several such systems, some of which are based on the principles and methods discussed above in the screening methods. This problem of the need for simpiicity has been addressed by Nicholls & Malcolm (1989) and Landegren et al (1988a).…”

Section: Mutation Detection Methods Where the Mutation Position Is Knmentioning

confidence: 99%

“…It was first used in diagnosis in 1983 to look for the presence of adenovirus (Ranki et al, 1983). Further applications to infectious organisms are reviewed in Nicholls & Malcolm (1989). The analysis of sequence variations was reported in 1985 (Langdale & Malcolm, 1985).…”

Section: Mutation Detection Methods Where the Mutation Position Is Knmentioning

confidence: 99%

Detection of single base changes in nucleic acids

Cotton

1989

Biochemical Journal

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“…Detection of amplified products on the dipstick membrane was achieved by sandwich nucleic acid hybridization (23); that is, by the hybridization of the target with two target-specific oligonucleotide probes, one for capture and one for detection. The low sensitivity of dipstick assays was overcome with the use of detector probes labeled with multiple hapten moieties for signal amplification.…”

Section: Viral Standards and Clinical Specimensmentioning

confidence: 99%

Simultaneous Visual Detection of Multiple Viral Amplicons by Dipstick Assay

et al. 2005

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A sensitive, simple, and instrument-independent method for the visual detection and identification of multiple nucleic acid amplicons by dipstick has been developed. This method is based on nucleic acid hybridization on the dipstick membrane and a signal amplification system to allow visual detection. The importance of nucleic acid testing (NAT) has become increasingly evident during the last decade for many purposes, such as diagnosing viral infections, monitoring antiviral therapy, and improving the safety of blood supplies. NAT combines the advantages of direct and highly sequence-specific detection of the genome of an infectious agent with an analytic sensitivity that is several orders of magnitude greater than that of antigen detection or virus isolation methods. NAT also reduces the risks of viral transmission during the period between infection and seroconversion (34), of infection with immunovariant viruses (20), of immunosilent carriage (6), and of occult carriage (3, 34). Unfortunately, a high investment cost for instruments, a high running cost for reagents, a lack of maintenance support, complex and often labor-intensive procedures that require highly trained personnel, and the need for cold-chain transport and storage of reagents render NAT unaffordable in the settings where it is needed most. These settings include predominantly the resource-limited countries of Africa, Asia, and Latin America with high prevalence of infectious diseases. Consequently, two complementary approaches to lower the costs of NAT have been proposed: pool testing for large-scale screening (26,35) and the development of multiplex assays for the simultaneous detection of several infectious agents (11).Multiplex assays have recently been developed for hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type 1 (HIV-1), which are three blood-borne pathogenic viruses of major epidemiological and clinical importance (9,19,21,22). The AMPLINAT MPX assay (21) and the multiplex real-time quantitative reverse transcription (RT)-PCR assay described by Candotti et al. (9) are based on TaqMan fluorescence technology, whereas the transcriptionmediated amplification-based Procleix Ultrio assay (19) relies on chemiluminescence technology for detection. NAT-based assays consist of three basic steps: extraction of nucleic acid, genome amplification mediated by a procedure such as PCR or RT-PCR, and amplicon detection. The first two steps can be performed with relatively inexpensive equipment, including a centrifuge, a water bath, and a thermal cycler. In contrast, the last step requires either time-consuming and labor-intensive electrophoretic separation or expensive equipment to detect fluorescence or chemiluminescence signals. The rapid visual detection of nucleic acid hybridization on a test strip (dipstick) would offer a simple and cost-effective alternative to these highly complex and instrument-dependent commercial methods. Serological testing performed with rapid dipstick tests is instrument independent ...

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Nucleic acid analysis by sandwich hybridization

Cited by 38 publications

References 108 publications

Chronic Blockade of Nitric Oxide Synthesis Reduces Adiposity and Improves Insulin Resistance in High Fat-Induced Obese Mice

Chronic Blockade of Nitric Oxide Synthesis Reduces Adiposity and Improves Insulin Resistance in High Fat-Induced Obese Mice

Detection of single base changes in nucleic acids

Simultaneous Visual Detection of Multiple Viral Amplicons by Dipstick Assay

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