Algorithms for compact letter displays: Comparison and evaluation

Multiple pairwise comparisons are one of the most frequent tasks in applied statistics. In this context, letter displays may be used for a compact presentation of results of multiple comparisons. A heuristic previously proposed for this task is compared with two new algorithmic approaches. The latter rely on the equivalence of computing compact letter displays to computing clique covers in graphs, a problem that is well-studied in theoretical computer science. A thorough discussion of the three approaches aims to give a comparison of the algorithms’ advantages and disadvantages. The three algorithms are compared in a series of experiments on simulated and real data, e.g., using data from wheat and triticale yield trials.     

A strategy of plasma protein quantitation by selective reaction monitoring of an intact protein

Immunoassays are used extensively in the quantitative analysis of proteins in plasma, urine, and other biological matrixes to support preclinical and clinical studies. Although immunoassays are both sensitive and rapid, difficulties during development of these assays are compounded by the need to have a specific antibody or antigen to the protein of interest. Furthermore, calibration curves of immunoassays are inherently nonlinear, and the technique often detects many structurally related components in addition to the analyte of interest. We have developed a novel strategy of analyzing protein concentrations in plasma by utilizing 96-well solid-phase extraction and LC-MS/MS detection of the intact protein. This strategy has been successfully applied in method development and assay validation of quantitatively analyzing protein rK5 concentrations in monkey plasma samples. Additional techniques such as precolumn regeneration and column heating were also incorporated into the assay. Total run time for each sample was approximately 15 min. An LLOQ of 99.2 ng/mL from a sample volume of 50 microL, corresponding to only 380 fmol (3.97 ng) of the rK5 analyte being injected onto the analytical column (assuming 100% extraction recovery), was obtained. The validated linear dynamic range was between 99.2 and 52 920.0 ng/mL, with a correlation coefficient (r(2)) ranging from 0.9972 and 0.9994. The intraassay CV for this assay was between 0.6 and 3.8%, and the interassay CV was between 1.7 and 3.2%. Interassay mean accuracies were between 101.5 and 104.7%. The assay has proven rugged and specific and has been employed to generate data in support of preclinical studies. This strategy for rK5 assay could be used for the development of bioanalytical assays to provide preclinical and clinical support for other protein drug candidates and, furthermore, for the validation of biomarkers discovered from proteomic research.     

The effects of PH, Ionic concentration and ionic species of dissolution media on the release rates of quinidine gluconate sustained release dosage forms

The dissolution profiles of some extended release quinidine gluconate products were shown to be dependent on several dissolution medium variables. It was shown that, for a quinidine gluconate wax matrix tablet, the dissolution rate has an unexpected inverse relationship to the solubility as a function of pH. The dissolution rate is also affected by the concentration of chloride ion present in the dissolution medium apparently due to the inhibition of disintegration. It was found that the nature of the anion (inorganic vs. organic) of the dissolution buffer plays a significant role in the release of the drug. Salts of inorganic acids used as part of the buffer system lower the rate of release of quinidine gluconate from wax matrix tablets through an inhibition of disintegration. On the other hand, buffer salts of organic acids do not have any appreciable effect on the disintegration or dissolution of these tablets. Since the concentration of chloride ion in both gastric and intestinal juices is approximately 0.1M, this suggests that the use of a dissolution medium containing chloride ion represents the more appropriate approach when an in-vivo/in-vitro correlation is desired. For all the quinidine gluconate controlled release formulations studied, mechanisms of release are of at least two different types. This makes the selection of a single dissolution medium for in-vivo/in-vitro correlations either improbable or impossible.     

Single-molecule assays of calmodulin target binding detected with a calmodulin energy-transfer construct

We have detected single-molecule binding interactions of a target peptide with the calcium-signaling protein calmodulin (CaM) immobilized in an agarose gel, and we have demonstrated the application of a single-molecule binding assay to measure the binding strength of CaM with the CaM-binding domain of calmodulin-dependent protein kinase II (CaMKII). The results demonstrate the potential for ultrasensitive assays of CaM-target interactions and the measurement of a picomolar dissociation constant. To detect single-molecule protein interactions, single-molecule assays require that the analyte molecule be confined to the focal spot of the objective for the time scale of the measurement. We demonstrate the deleterious effect of surface immobilization on CaM. As an alternative to surface immobilization, we have constructed a CaM/maltose binding protein fusion protein, which renders CaM translationally immobile in a low weight percent agarose gel. The target binding functionality of CaM assayed in agarose gels is in good agreement with solution assays. The utility of the construct for detecting interactions with CaM targets was demonstrated in a single-molecule assay of binding interactions of MBP-CaM with the CaMKII CaM-binding domain peptide. A value of 103 +/- 35 pM for the dissociation constant of this interaction was determined by simple counting of fluorescent molecules.     

A behavioral change-based approach to energy efficiency in a manufacturing plant

In the realm of industrial energy efficiency studies, very little research has been done to understand the barriers and opportunities to influence behavior of production workers and the corresponding energy-saving potentials. This paper analyzes a case study of Volvo Construction Equipment AB in Braås, Sweden (VCE Braås), that has reduced its relative idle electricity use by more than 10 percentage points during 2013–2016 by implementing a strategy of changing everyday behavior of production workers. The results based on interviews with actors involved in the energy efficiency project showed that a concrete goal, the employment of a fulltime operational leader who earlier worked as a production worker at VCE Braås, and the involvement of both the leadership and employees in project management were key to the success of the project.     

Gold Nanoparticle‐Based Sensors Activated by External Radio Frequency Fields

A novel molecular beacon (a nanomachine) is constructed that can be actuated by a radio frequency (RF) field. The nanomachine consists of the following elements arranged in molecular beacon configuration: a gold nanoparticle that acts both as quencher for fluorescence and a localized heat source; one reporter fluorochrome, and; a piece of DNA as a hinge and recognition sequence. When the nanomachines are irradiated with a 3 GHz RF field the fluorescence signal increases due to melting of the stem of the molecular beacon. A control experiment, performed using molecular beacons synthesized by substituting the gold nanoparticle by an organic quencher, shows no increase in fluorescence signal when exposed to the RF field. It may therefore be concluded that the increased fluorescence for the gold nanoparticle‐conjugated nanomachines is not due to bulk heating of the solution, but is caused by the presence of the gold nanoparticles and their interaction with the RF field; however, existing models for heating of gold nanoparticles in a RF field are unable to explain the experimental results. Due to the biocompatibility of the construct and RF treatment, the nanomachines may possibly be used inside living cells. In a separate experiment a substantial increase in the dielectric losses can be detected in a RF waveguide setup coupled to a microfluidic channel when gold nanoparticles are added to a low RF loss liquid. This work sheds some light on RF heating of gold nanoparticles, which is a subject of significant controversy in the literature.     

Variable time-delay estimation for anesthesia control during intensive care

The presence of artifacts plays a crucial role in automatic sedation systems and may introduce variable time delays (TDs) in the closed-loop-control structures. This paper presents a successful procedure to estimate the varying TD of the bispectral index (BIS) monitor used in closed-loop control during intensive care. The TD estimation (TDE) is based on the cross-correlation analysis technique and the method is validated with real measured signals of propofol and BIS. Extended prediction self-adaptive control is used in combination with a Smith predictor to reduce the computational burden imposed by the variable TD. The conclusion is that an online TDE of the BIS monitor improves the performance of the closed-loop system for reference tracking, disturbance rejection, and overall stability.     

Pre-natal social stress and post-natal pain affect the developing pig reproductive axis

This study assessed the effect of pre-natal social stress and post-natal pain on the reproductive development of young (approximately day 40) pigs. Male pigs carried by sows that were stressed by mixing with unfamiliar older sows for two 1-week periods during mid-pregnancy had lower plasma testosterone (0.54 vs 0.86 ng/ml, S.E.D.=0.11; P=0.014) and oestradiol (E(2); 22.9 vs 38.7 pg/ml, S.E.D.=7.80; P=0.021) concentrations compared with males carried by unstressed control sows. Although there was no effect of pre-natal stress on female E(2) concentrations, female pigs carried by stressed sows had fewer primordial ovarian follicles (log -4.32/μm(2) vs -4.00/μm(2), s.e.d.=0.136; P=0.027). Tail amputation on day 3 after birth reduced E(2) concentrations in female (4.78 vs 6.84 pg/ml, s.e.d.=0.86; P=0.03) and in male (25.6 vs 34.9 pg/ml, S.E.D.=3.56; P=0.021) pigs and reduced both testis weight (0.09% of body weight vs 0.10% of body weight, S.E.D.=0.003; P=0.01) and the percentage of proliferating Leydig cells (1.97 vs 2.12, S.E.D.=0.114; P=0.036) compared with sham-amputated littermate controls. There was a significant (P=0.036) interaction between the effects of pre-natal stress and post-natal pain on testicular expression of the steroidogenic enzyme 17α-hydroxylase, such that amputation increased expression in pigs born to control sows, but reduced expression in animals born to stressed sows. This study shows that stressful procedures associated with routine animal husbandry can disrupt the developing reproductive axis.