All-Trans Retinoic Acid Induces DU145 Cell Cycle Arrest through Cdk5 Activation

All-trans retinoic acid (ATRA), the active form of vitamin A, plays an important role in the growth arrest of numerous types of cancer cells. It has been indicated that cyclin-dependent kinase 5 (Cdk5) activity can be affected by ATRA treatment. Our previous results demonstrate the involvement of Cdk5 in the fate of prostate cancer cells. The purpose of this study is to examine whether Cdk5 is involved in ATRA-induced growth arrest of the castration-resistant cancer cell line DU145 through up-regulating Cdk inhibitor protein, p27

Selection of DDX5 as a novel internal control for Q-RT-PCR from microarray data using a block bootstrap re-sampling scheme

<p>Abstract</p> <p>Background</p> <p>The development of microarrays permits us to monitor transcriptomes on a genome-wide scale. To validate microarray measurements, quantitative-real time-reverse transcription PCR (Q-RT-PCR) is one of the most robust and commonly used approaches. The new challenge in gene quantification analysis is how to explicitly incorporate statistical estimation in such studies. In the realm of statistical analysis, the various available methods of the probe level normalization for microarray analysis may result in distinctly different target selections and variation in the scores for the correlation between microarray and Q-RT-PCR. Moreover, it remains a major challenge to identify a proper internal control for Q-RT-PCR when confirming microarray measurements.</p> <p>Results</p> <p>Sixty-six Affymetrix microarray slides using lung adenocarcinoma tissue RNAs were analyzed by a statistical re-sampling method in order to detect genes with minimal variation in gene expression. By this approach, we identified <it>DDX5 </it>as a novel internal control for Q-RT-PCR. Twenty-three genes, which were differentially expressed between adjacent normal and tumor samples, were selected and analyzed using 24 paired lung adenocarcinoma samples by Q-RT-PCR using two internal controls, <it>DDX5 </it>and <it>GAPDH</it>. The percentage correlation between Q-RT-PCR and microarray were 70% and 48% by using <it>DDX5 </it>and <it>GAPDH </it>as internal controls, respectively.</p> <p>Conclusion</p> <p>Together, these quantification strategies for Q-RT-PCR data processing procedure, which focused on minimal variation, ought to significantly facilitate internal control evaluation and selection for Q-RT-PCR when corroborating microarray data.</p

Toxicity of Transition Metal Complex-based Nanophotoswitches in Retina

Purpose: Nanophotoswitches (NPSs) based on transition metal complexes offer a new tool for optical stimulation of neural activity in photoreceptor degenerated retina. We previously reported robust light-elicited neural activity in degenerate retinae exposed to ruthenium bipyridine based NPSs (Rubpy-C17) and its iridium analog (Irbpy-C17). Irbpy-C17 was developed as an alternative to Rubpy-C17 for the biosafety properties of the iridium complexes. Here we present a study of the toxicity of both NPSs in rodent retinae. Methods: Toxicity of Rubpy-C17 was tested in wildtype C57BL/6J mice and Irbpy-C17 in wildtype Long Evans rats. Animals were intravitreally injected with the test molecules (up to 50 µM) and sacrificed at different time points post injection: 3, 7, 14 and 28 days, respectively. Retinae were obtained, fixed and sliced for histological analysis immediately after animal euthanization. H&E staining was performed to examine morphological integrity of retina and TUNEL staining performed to detect apoptosis of retinal cells. For comparison, Ru(bpy)3]Cl2 injection and sham surgery were included for control. Results: H&E staining revealed no detectable sign of morphological or structural changes in the retinae after prolonged exposure to either Rubpy-C17 or Irbpy-C17 versus the control. There was no significant reduction in the thickness of different nuclear and plexiform retinal layers or the density of retinal neurons (p<0.05), nor was there evidence of significant aggregation of immune cells. TUNEL staining showed minimal occurrence of cell apoptosis in the NPS treated retinae, similar to the control (p<0.05). No longitudinal changes in either the morphology or the cell apoptosis was observed with the post injection time. Conclusions: Overall our data did not find ocular toxicity associated with either the ruthenium or the iridium based NPSs within the concentration range tested. The results obtained with both complexes are similar to that obtained with the control molecule [Ru(bpy)3]Cl2, which lacks a membrane-anchoring 17 carbon chain attached to the bipyridine group, indicating that the inclusion of the carbon chain did not enable NPSs entry into the cells, nor did it cause apoptotic response. The present study provides new evidence of biosafety of our NPSs in rodent retinae, further encouraging developing NPS-based molecular retinal prosthesis to potentially restore high-acuity prosthetic vision in the blind

A Study of Antibioactivity of Nanosilver Colloid and Silver Ion Solution

The colloidal silver solution was successfully prepared in dielectric fluid by using electrical spark discharge (ESD) without any surfactants. It does not require the toxic chemical agents in the process, which may affect the effectiveness of nanosilver colloid as an antibacterial agent. Nanocolloidal silver produced by ESD is characterized as low cost, zero environmental pollution, continuous, and rapid mass production process. In order to test the effect of antibioactivity, nanosilver dough was tested; the silver nanofluid was prepared by ESD machine, made into dough at different concentrations, and fermented for three hours in order to observe changes in the diameter of the dough. The results showed that the effect of effectiveness of nanosilver at the concentration of 100 ppm was weak, whereas the effect of 60 ppm silver ion (100 ppm AgNO3) was significant, as the dissociation rate of silver ion concentration correlates to the antibioactivity

Comparison of Antimicrobial Susceptibility Testing of Isolates from Blood Cultures by Direct Inoculation Method and PHOENIX

Background: Bloodstream infection (BSI) is an important cause of serious morbidity and mortality for hospitalized patients. Empirically Gram stain of bacteria gives the first clue for the etiology of infection and medical treatment. But the delayed treatment on 1 or 2 days after phenotypic identification and drug susceptibility testing may cause potential danger to patients. Rapid drug susceptibility testing can provide earlier information to guide treatment and in less time than bacterial culture and sensitivity testing, for antibiotics therapy. Methods: In this study, we excluded samples of polymicrobial bacteremia. We collected isolates from 815 infection episodes caused by Escherichia coli (57%), Klebsiella pneumoniae (20.16%), Enterobacter cloacae (6%), Ps.aeruginosa (9.1%), Stenotrophomonas maltophilia (3.1%), and Acinetobacter baumannii (3.1%) in a 10-month period. We identified those bacteria with direct susceptibility test with the use of Phoenix100 (BD) during a 10-month period. Results: The results of direct susceptibility were concordant (99%-100%) with those obtained from Phoenix100. Conclusion: These results have the potential to guide clinicians to initiate an early antimicrobial therapy in febrile patients with sepsis shock

Two-tiered Online Optimization of Region-wide Datacenter Resource Allocation via Deep Reinforcement Learning

This paper addresses the important need for advanced techniques in continuously allocating workloads on shared infrastructures in data centers, a problem arising due to the growing popularity and scale of cloud computing. It particularly emphasizes the scarcity of research ensuring guaranteed capacity in capacity reservations during large-scale failures. To tackle these issues, the paper presents scalable solutions for resource management. It builds on the prior establishment of capacity reservation in cluster management systems and the two-level resource allocation problem addressed by the Resource Allowance System (RAS). Recognizing the limitations of Mixed Integer Linear Programming (MILP) for server assignment in a dynamic environment, this paper proposes the use of Deep Reinforcement Learning (DRL), which has been successful in achieving long-term optimal results for time-varying systems. A novel two-level design that utilizes a DRL-based algorithm is introduced to solve optimal server-to-reservation assignment, taking into account of fault tolerance, server movement minimization, and network affinity requirements due to the impracticality of directly applying DRL algorithms to large-scale instances with millions of decision variables. The paper explores the interconnection of these levels and the benefits of such an approach for achieving long-term optimal results in the context of large-scale cloud systems. We further show in the experiment section that our two-level DRL approach outperforms the MIP solver and heuristic approaches and exhibits significantly reduced computation time compared to the MIP solver. Specifically, our two-level DRL approach performs 15% better than the MIP solver on minimizing the overall cost. Also, it uses only 26 seconds to execute 30 rounds of decision making, while the MIP solver needs nearly an hour

Biochemical Properties and Potential Applications of Recombinant Leucine Aminopeptidase from Bacillus kaustophilus CCRC 11223

Experiments were carried out to investigate the effects of various factors on the activity and conformation of recombinant leucine aminopeptidase of Bacillus kaustophilus CCRC 11223 (BkLAP) and potential utilization of BkLAP in the hydrolysis of anchovy protein. Optimal temperature and pH of BkLAP were 70 °C and 8.0 in potassium-phosphate buffer, respectively, and the activity was strongly stimulated by Ni2+, followed by Mn2+ and Co2+. Conformational studies via circular dichroism spectroscopy indicated that various factors could influence the secondary structure of BkLAP to different extents and further induce the changes in enzymatic activity. The secondary structure of BkLAP was slightly modified by Ni2+ at the concentration of 1×10−4 M, however, significant changes on the secondary structures of the enzyme were observed when Hg2+ was added to the concentration of 1×10−4 M. The potential application of BkLAP was evaluated through combination with the commercial or endogenous enzyme to hydrolysis the anchovy protein. Results showed that combining the BkLAP with other enzymes could significantly increase the degree of hydrolysis and amino acid component of hydrolysate. In this regard, BkLAP is a potential enzyme that can be used in the protein hydrolysate industry

Toxicity of Transition Metal Complex-based Nanophotoswitches in Retina

Purpose: Nanophotoswitches (NPSs) based on transition metal complexes offer a new tool for optical stimulation of neural activity in photoreceptor degenerated retina. We previously reported robust light-elicited neural activity in degenerate retinae exposed to ruthenium bipyridine based NPSs (Rubpy-C17) and its iridium analog (Irbpy-C17). Irbpy-C17 was developed as an alternative to Rubpy-C17 for the biosafety properties of the iridium complexes. Here we present a study of the toxicity of both NPSs in rodent retinae. Methods: Toxicity of Rubpy-C17 was tested in wildtype C57BL/6J mice and Irbpy-C17 in wildtype Long Evans rats. Animals were intravitreally injected with the test molecules (up to 50 µM) and sacrificed at different time points post injection: 3, 7, 14 and 28 days, respectively. Retinae were obtained, fixed and sliced for histological analysis immediately after animal euthanization. H&E staining was performed to examine morphological integrity of retina and TUNEL staining performed to detect apoptosis of retinal cells. For comparison, Ru(bpy)3]Cl2 injection and sham surgery were included for control. Results: H&E staining revealed no detectable sign of morphological or structural changes in the retinae after prolonged exposure to either Rubpy-C17 or Irbpy-C17 versus the control. There was no significant reduction in the thickness of different nuclear and plexiform retinal layers or the density of retinal neurons (p<0.05), nor was there evidence of significant aggregation of immune cells. TUNEL staining showed minimal occurrence of cell apoptosis in the NPS treated retinae, similar to the control (p<0.05). No longitudinal changes in either the morphology or the cell apoptosis was observed with the post injection time. Conclusions: Overall our data did not find ocular toxicity associated with either the ruthenium or the iridium based NPSs within the concentration range tested. The results obtained with both complexes are similar to that obtained with the control molecule [Ru(bpy)3]Cl2, which lacks a membrane-anchoring 17 carbon chain attached to the bipyridine group, indicating that the inclusion of the carbon chain did not enable NPSs entry into the cells, nor did it cause apoptotic response. The present study provides new evidence of biosafety of our NPSs in rodent retinae, further encouraging developing NPS-based molecular retinal prosthesis to potentially restore high-acuity prosthetic vision in the blind