AAA ATPases as therapeutic targets: Structure, functions, and small-molecule inhibitors

ATPases Associated with Diverse Cellular Activity (AAA ATPase) are essential enzymes found in all organisms. They are involved in various processes such as DNA replication, protein degradation, membrane fusion, microtubule serving, peroxisome biogenesis, signal transduction, and the regulation of gene expression. Due to the importance of AAA ATPases, several researchers identified and developed small-molecule inhibitors against these enzymes. We discuss six AAA ATPases that are potential drug targets and have well-developed inhibitors. We compare available structures that suggest significant differences of the ATP binding pockets among the AAA ATPases with or without ligand. The distances from ADP to the His20 in the His-Ser-His motif and the Arg finger (Arg353 or Arg378) in both RUVBL1/2 complex structures bound with or without ADP have significant differences, suggesting dramatically different interactions of the binding site with ADP. Taken together, the inhibitors of six well-studied AAA ATPases and their structural information suggest further development of specific AAA ATPase inhibitors due to difference in their structures. Future chemical biology coupled with proteomic approaches could be employed to develop variant specific, complex specific, and pathway specific inhibitors or activators for AAA ATPase proteins

Development and Validation of the Parents\u27 Perceived Self-Efficacy to Manage Children\u27s Internet Use Scale for Parents of Adolescents with Attention-Deficit/Hyperactivity Disorder

Background and aims: This study developed and validated the Parents’ Perceived Self-Efficacy to Manage Children’s Internet Use Scale (PSMIS) in the parents of children with attention-deficit/hyperactivity disorder (ADHD). Methods: In total, 231 parents of children with ADHD were invited to complete the PSMIS, followed by the Chen Internet Addiction Scale and the short version of Swanson, Nolan, and Pelham, Version IV Scale – Chinese version for analyzing Internet addiction severity and ADHD symptoms, respectively. Results: The results of exploratory and confirmatory factor analyses confirmed the four-factor structure of the 18-item PSMIS. The significant difference in the levels of parents’ perceived self-efficacy between the parents of children with and without Internet addiction supported the criterion-related validity of the PSMIS. The internal consistency and 1-month test–retest reliability were acceptable. Conclusion: The results indicate that the PSMIS has acceptable validity and reliability and can be used for measuring parents’ perceived self-efficacy to manage children’s Internet use among parents of children with ADHD

Osteomyelitis of Multiple Lumbar Vertebrae Associated with Infected Aortic Aneurysm: A Case Report

A 73-year-old male patient presented with a pulsating abdominal mass and intractable low back pain for several days. Magnetic resonance imaging revealed an infected abdominal aortic aneurysm invading the second, third, and fourth lumbar vertebrae. He underwent radical debridement of the infected aneurysm with reconstruction using vascular bypass, partial corpectomy of the L2 to L4 vertebrae, anterior reconstruction with autogenous fibular shaft, and posterior instrumentation with posterolateral fusion. Culture of the necrotic tissues grew oxacillin-resistant Staphylococcus aureus. He received intravenous vancomycin infusion for 4 weeks and oral ciprofloxacin for 6 months postoperatively. After a 15-month follow-up, no apparent signs of further infection were noted. C-reactive protein and erythrocyte sedimentation rate returned to normal during follow-up. No neurologic symptoms other than mild low back soreness were noted. The stability of the lumbar spine was maintained using long segment reconstruction with autogenous fibula shaft and posterior instrumentation along with posterolateral fusion. Infected aortic aneurysm with vertebral osteomyelitis is a rare clinical entity. Prompt diagnosis and adequate treatment are essential

New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation

AbstractObjectiveSignal transducer and activator of transcription (STAT)1 is a key tumor suppressor, which is always methylated in a variety of human cancers. However, nonspecific primers for the detection of specific promoter hypermethylation of STAT1 gene can lead to false-positive or false-negative results for gene methylation.Materials and MethodsWe designed new primers for the detection of STAT1 methylation and compared the sensitivities and specificities of these new primers with prior published primers by methylation-specific polymerase chain reaction (PCR) from ovarian clear cell carcinomas. The mRNA expression levels of STAT1 in these cancerous tissues were also evaluated by reverse-transcriptase PCR and correlated with the results of promoter methylation of STAT1 gene.ResultsNine (39%) of the 23 samples detected by the new primers and 13 samples (56%) detected by prior published primers showed STAT1 methylation. A direct DNA sequencing test revealed that four of the 13 samples (30.8%) showed false positivity for STAT1 methylation using the prior published primers. In contrast, none of the nine samples was false-positive for the detection of STAT1 methylation using the new primers. The new primers for the detection of STAT1 methylation showed 100% specificity and 100% sensitivity without false positivity.ConclusionSpecific primers for methylation-specific PCR are mandatory for the accurate detection of STAT1 gene methylation. Besides, specific primers can generate correct interpretation of STAT1 gene methylation, and its correlation with the clinicopathological characteristics and outcome of cancer patients

High-throughput avian molecular sexing by SYBR green-based real-time PCR combined with melting curve analysis

<p>Abstract</p> <p>Background</p> <p>Combination of <it>CHD </it>(chromo-helicase-DNA binding protein)-specific polymerase chain reaction (PCR) with electrophoresis (PCR/electrophoresis) is the most common avian molecular sexing technique but it is lab-intensive and gel-required. Gender determination often fails when the difference in length between the PCR products of <it>CHD-Z </it>and <it>CHD-W </it>genes is too short to be resolved.</p> <p>Results</p> <p>Here, we are the first to introduce a PCR-melting curve analysis (PCR/MCA) to identify the gender of birds by genomic DNA, which is gel-free, quick, and inexpensive. <it>Spilornis cheela hoya </it>(<it>S. c. hoya</it>) and <it>Pycnonotus sinensis </it>(<it>P. sinensis</it>) were used to illustrate this novel molecular sexing technique. The difference in the length of <it>CHD </it>genes in <it>S. c. hoya </it>and <it>P. sinensis </it>is 13-, and 52-bp, respectively. Using Griffiths' P2/P8 primers, molecular sexing failed both in PCR/electrophoresis of <it>S. c. hoya </it>and in PCR/MCA of <it>S. c. hoya </it>and <it>P. sinensis</it>. In contrast, we redesigned sex-specific primers to yield 185- and 112-bp PCR products for the <it>CHD-Z </it>and <it>CHD-W </it>genes of <it>S. c. hoya</it>, respectively, using PCR/MCA. Using this specific primer set, at least 13 samples of <it>S. c. hoya </it>were examined simultaneously and the Tm peaks of <it>CHD-Z </it>and <it>CHD-W </it>PCR products were distinguished.</p> <p>Conclusion</p> <p>In this study, we introduced a high-throughput avian molecular sexing technique and successfully applied it to two species. This new method holds a great potential for use in high throughput sexing of other avian species, as well.</p

Toward Inter-Connection on OpenFlow Research Networks

With the advance of Future Internet technologies, many research issues andideas are growing fast in recent years. In the field of network virtualization, softwaredefined network becomes a common topic on network research. In Taiwan, manyinstitutes and laboratories of universities already built their bench-scale testbed forresearch and educational use with OpenFlow protocol. As time goes by, stitchingexperimental networks is a growing trend to fulfill requirements for large scaleemulation. Hence, this paper revealed a progressing deployment which connectsdifferent experimental networks with centralized control policy. The objective is tobuild an integrated research network with a proposed solution which utilizesOpenFlow protocol to deal with the inter-connections. With a centralized controllerand implemented architecture, the deployment not only solves the limitation of VLANtag number in network but also improves the flexibility of configuration. This designcould be a solution for the realistic constraints of network environment in Taiwan, andit also supports the possibility of stitching regional experimental networks fornetworking research

Structural and cognitive deficits in chronic carbon monoxide intoxication: a voxel-based morphometry study

BACKGROUND: Patients with carbon monoxide (CO) intoxication may develop ongoing neurological and psychiatric symptoms that ebb and flow, a condition often called delayed encephalopathy (DE). The association between morphologic changes in the brain and neuropsychological deficits in DE is poorly understood. METHODS: Magnetic resonance imaging and neuropsychological tests were conducted on 11 CO patients with DE, 11 patients without DE, and 15 age-, sex-, and education-matched healthy subjects. Differences in gray matter volume (GMV) between the subgroups were assessed and further correlated with diminished cognitive functioning. RESULTS: As a group, the patients had lower regional GMV compared to controls in the following regions: basal ganglia, left claustrum, right amygdala, left hippocampus, parietal lobes, and left frontal lobe. The reduced GMV in the bilateral basal ganglia, left post-central gyrus, and left hippocampus correlated with decreased perceptual organization and processing speed function. Those CO patients characterized by DE patients had a lower GMV in the left anterior cingulate and right amygdala, as well as lower levels of cognitive function, than the non-DE patients. CONCLUSIONS: Patients with CO intoxication in the chronic stage showed a worse cognitive and morphologic outcome, especially those with DE. This study provides additional evidence of gray matter structural abnormalities in the pathophysiology of DE in chronic CO intoxicated patients

Antioxidant activity and growth inhibition of human colon cancer cells by crude and purified fucoidan preparations extracted from Sargassum cristaefolium

AbstractFucose-containing sulfated polysaccharides, also termed “fucoidans”, which are known to possess antioxidant, anticoagulant, anticancer, antiviral, and immunomodulating properties, are normally isolated from brown algae via various extraction techniques. In the present study, two methods (SC1 and SC2) for isolation of fucoidan from Sargassum cristaefolium were compared, with regard to the extraction yields, antioxidant activity, and inhibition of growth of human colon cancer cells exhibited by the respective extracts. SC1 and SC2 differ in the number of extraction steps and concentration of ethanol used, as well as the obtained sulfated polysaccharide extracts, namely, crude fucoidan preparation (CFP) and purified fucoidan preparation (PFP), respectively. Thin layer chromatography, Fourier transform infrared analysis, and measurements of fucose and sulfate contents revealed that the extracts were fucoidan. There was a higher extraction yield for CFP, which contained less fucose and sulfate but more uronic acid, and had weaker antioxidant activity and inhibition of growth in human colon cancer cells. In contrast, there was a lower extraction yield for PFP, which contained more fucose and sulfate but less uronic acid, and had stronger antioxidant activity and inhibition of growth in human colon cancer cells. Thus, since the difference in bioactive activities between CFP and PFP was not remarkable, the high extraction yield of SC1 might be favored as a method in industrial usage for extracting fucoidan