Use of cumulative meta-analysis in the design, monitoring, and final analysis of a clinical trial: a case study

From 1983 to 1987, the Department of Veterans Affairs (DVA) Cooperative Studies Program (CSP) conducted a multicenter clinical trial (CSP #207) to determine whether four different antiplatelet regimens compared to placebo could prevent the occlusion of grafts following coronary artery bypass surgery. The study showed that all of the active regimens tended to be better than placebo and that the three regimens containing aspirin were statistically significantly better. A cumulative meta-analysis of 12 trials performed shortly before the end of CSP #207 raised the issue as to whether the meta-analysis, if done earlier, would have changed the conduct of the trial. At the start of the planning period, one trail of size n = 37 had been published with a nonsignificant odds ratio (OR) of 0.74 (95% CI: 0.18, 3.12). At the time that CSP # 207 was approved by the DVA Cooperative Studies Evaluation Committee, two trials had been published (cumulative n = 150, OR = 0.44, 95% CI 0.19, 0.99). At the time patient intake started, five trials showed cumulative n = 769, OR = 0.42, 95% CI = 0.26, 0.68. Although the first 6-month CSP #207 progress report showed no treatment effect, by the time of the 12-month review by the Data Monitoring Board (DMB) a trend was developing in favor of active treatment. If the results of the meta-analysis had been available to the DMB at that time, conceivably the Board would have recommended stopping the placebo arm because of a convincing treatment effect based on the totality of the evidence. Cumulative meta-analysis could be useful as an adjunct in the planning, conduct, and final analysis of a clinical trial. It could also be used as one piece of evidence in the monitoring of the ongoing phase of a trial.     

Continuous-wave operation of GaInAs-AlGaAsSb quantum cascade lasers

We report on the demonstration of continuous-wave (CW) operation of GaInAs-AlGaAsSb quantum cascade (QC) lasers. By placing a 2.5-/spl mu/m-thick gold layer on both sides of the laser ridge to extract heat from the active region in the lateral direction, together with mounting the device epilayer down, we have achieved CW operation of GaInAs-AlGaAsSb QC lasers composed of 25 stages of active/injection regions. The maximum CW operating temperature of the lasers is 94 K, and the emission wavelength is around /spl lambda//spl sim/4.65 /spl mu/m. For a device with the size of 10/spl times/2000 /spl mu/m/sup 2/, the CW optical output power per facet is 13 mW at 42 K and 4 mW at 94 K. The CW threshold current density is 1.99 kA/cm/sup 2/ at 42 K, and 2.08 kA/cm/sup 2/ at 94 K, respectively.     

Learning and chronobiological regulation following experimental damage to the coronary-artery wall of albino rats]

Forty male albino rats were used to study the relationship between experimentally (concentrated hydrochloric acid) produced local structural changes of the coronary artery wall and central nervous information processing and chronobiological events, respectively. The alterations of the coronary arteries caused by this treatment did not produce infarction but initiated on the whole length of the arterial branch proliferative processes that are complete within 12 weeks following the intervention. Central nervous information processing and chronoregulatory processes showed, immediately after exposure to hydrochloric acid, pronounced malfunctions followed by a stage of hyperreactivity (4 weeks following the damage). At 8 weeks from the intervention, the parameters of information processing and chronobiological events of the CNS again equalled the control values. Since the central nervous functions were back to normal earlier than the structural alterations, the results are interpreted as reflecting CNS-controlled interactions between the complex of visceral afference and efference, the regulation of which is determined by the CNS after coronary damage in such a way as to restore the adaptational capacity of the organism by a predominance of sanogenetic processes.     

High-throughput data analysis for detecting and identifying differences between samples in GC/MS-based metabolomic analyses

In metabolomics, the objective is to identify differences in metabolite profiles between samples. A widely used tool in metabolomics investigations is gas chromatography-mass spectrometry (GC/MS). More than 400 compounds can be detected in a single analysis, if overlapping GC/MS peaks are deconvoluted. However, the deconvolution process is time-consuming and difficult to automate, and additional processing is needed in order to compare samples. Therefore, there is a need to improve and automate the data processing strategy for data generated in GC/MS-based metabolomics; if not, the processing step will be a major bottleneck for high-throughput analyses. Here we describe a new semiautomated strategy using a hierarchical multivariate curve resolution approach that processes all samples simultaneously. The presented strategy generates (after appropriate treatment, e.g., multivariate analysis) tables of all the detected metabolites that differ in relative concentrations between samples. The processing of 70 samples took similar time to that of the GC/TOFMS analyses of the samples. The strategy has been validated using two different sets of samples: a complex mixture of standard compounds and Arabidopsis samples.     

The cyclophilin component of the unactivated estrogen receptor contains a tetratricopeptide repeat domain and shares identity with p59 (FKBP59)

Using a rapid single-step affinity chromatography procedure we have isolated the unactivated estrogen receptor from bovine uterus. Results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western analyses for protein extracts recovered from affinity chromatography of receptor cytosols, either preincubated or untreated with estradiol, suggest a component structure for the intact oligomeric receptor which includes hsp90, hsp70, p59, a 40-kDa cyclophilin-related protein, and an uncharacterized 22-kDa protein species. We have chemically determined the amino acid sequences of eight peptides derived from the 40-kDa component and now report the cloning and primary sequence of a cDNA encoding this protein, which is designated estrogen receptor-binding cyclophilin (ERBC). Homology analyses confirm that ERBC is a new member of the cyclophilin family and contains a C-terminal domain with significant sequence homology to an internal region of p59, a binding protein for the immunosuppressant FK506 (FKBP59). This conserved region includes a 3-unit tetratricopeptide repeat domain bounded at the C terminus by a putative calmodulin binding site. We propose that the tetratricopeptide repeat domain mediates the protein interaction properties of ERBC and p59. Both immunophilins may have important roles in receptor assembly and may represent a new category of ligand- and calcium-dependent modulators of protein function.     

Engineered superoxide dismutase monomers for superoxide biosensor applications

Because of its high reaction rate and specificity, the enzyme superoxide dismutase (SOD) offers great potential for the sensitive quantification of superoxide radicals in electrochemical biosensors. In this work, monomeric mutants of human Cu,Zn-SOD were engineered to contain one or two additional cysteine residues, which could be used to bind the protein to gold surfaces, thus making the use of promotor molecules unnecessary. Six mutants were successfully designed, expressed, and purified. All mutants bound directly to unmodified gold surfaces via the sulfur of the cysteine residues and showed a quasi-reversible, direct electron transfer to the electrode. Thermodynamic and kinetic parameters of the electron transfer were characterized and showed only slight variations between the individual mutants. For one of the mutants, the interaction with the superoxide radical was studied in more detail. For both partial reactions of the dismutation, an interaction between protein and radical could be shown. In an amperometric biosensorial approach, the SOD-mutant electrode was successfully applied for the detection of superoxide radicals. In the oxidation region, the electrode surpassed the sensitivity of the commonly used cytochrome c electrodes by approximately 1 order of magnitude while not being limited by interferences, but the electrode did not fully reach the sensitivity of dimeric Cu,Zn-SOD immobilized on MPA-modified gold.