Suitability of PCR Fingerprinting, Infrequent-Restriction-Site PCR, and Pulsed-Field Gel Electrophoresis, Combined with Computerized Gel Analysis, in Library Typing of Salmonella enterica Serovar Enteritidis

Strains of Salmonella enterica (n = 212) of different serovars and phage types were used to establish a library typing computerized system for serovar Enteritidis on the basis of PCR fingerprinting, infrequent-restriction-site PCR (IRS-PCR), or pulsed-field gel electrophoresis (PFGE). The rate of PCR fingerprinting interassay and intercenter reproducibility was low and was only increased when DNA samples were extracted at the same time and amplified with the same reaction mixtures. Reproducibility of IRS-PCR technique reached 100%, but discrimination was low (D = 0.52). The PFGE procedure showed an intercenter reproducibility value of 93.3%. The high reproducibility of PFGE combined with the previously determined high discrimination directed its use for library typing. The use of PFGE with enzymes XbaI, BlnI, and SpeI for library typing of serovar Enteritidis was assessed with GelCompar 4.0 software. Three computer libraries of PFGE DNA profiles were constructed, and their ability to recognize new DNA profiles was analyzed. The results obtained pointed out that the combination of PFGE with computerized analysis could be suitable in long-term epidemiological comparison and surveillance of Salmonella serovar Enteritidis, specially if the prevalence of genetic events that could be responsible for changes in PFGE profiles in this serovar was low.     

Structures of ribosomal subunits from Saccharomyces cerevisiae

The large (60S) and small (40S) ribosomal subunits were isolated from yeast (Saccharomyces cerevisiae), and preparations stained with uranyl acetate were imaged by transmission electron microscopy. Averages of three different projections of each subunit were obtained by computerized image processing with reproducible spatial resolutions approaching 3.0 and 3.5 nm, respectively. Similarities between the structures of these particles and those of other eukaryotic ribosomal subunits contribute to the development of a concensus model for the eukaryotic ribosome.     

Diabetic Ketoacidosis Critical Care Pathway Implementation: Incorporation into EMR Significantly Decreases Length of Stay

Objective: We discuss the implementation and outcomes of a diabetic ketoacidosis (DKA) critical care pathway (CCP) at a 462-bed teaching hospital.Methods: A multi-disciplinary team implemented a DKA CCP that was translated into 3 computerized physician order entry (CPOE) order sets corresponding to the phases of DKA care. Historical and postintervention data were obtained via automated queries of the electronic medical record (EMR) and further analyzed by manual chart review.Results: Average length of stay decreased from 104.3 to 72.9 hours (P = .0003) after implementation of a DKA CCP.Conclusion: Outcome data supports the use of a DKA CCP at our institution.Abbreviations:DKA = diabetic ketoacidosisCCP = critical care pathwayEMR = electronic medical recordCPOE = computerized physician order entryICD-9 = International Classification of Diseases, ninth revisionLoS = length of staySQL = standard query language     

Is Working Memory Training Effective? A Study in a School Setting

We tested the effectiveness of an intensive, on average 17-session, adaptive and computerized working-memory training program for improving performance on untrained, paper and pencil working memory tasks, standardized school achievement tasks, and teacher ratings of classroom behavior. Third-grade children received either a computerized working memory training for about 30 minutes per session (n = 156) or participated in regular classroom activities (n = 126). Results indicated strong gains in the training task. Further, pretest and posttest transfer measures of working memory and school achievement, as well as teacher ratings, showed substantial correlations with training task performance, suggesting that the training task captured abilities that were relevant for the transfer tasks. However, effect sizes of training-specific transfer gains were very small and not consistent across tasks. These results raise questions about the benefits of intensive working-memory training programs within a regular school context.     

Spectral Sensitivity Measured with Electroretinogram Using a Constant Response Method

A new method is presented to determine the retinal spectral sensitivity function S(λ) using the electroretinogram (ERG). S(λ)s were assessed in three different species of myomorph rodents, Gerbils (Meriones unguiculatus), Wistar rats (Ratus norvegicus), and mice (Mus musculus). The method, called AC Constant Method, is based on a computerized automatic feedback system that adjusts light intensity to maintain a constant-response amplitude to a flickering stimulus throughout the spectrum, as it is scanned from 300 to 700 nm, and back. The results are presented as the reciprocal of the intensity at each wavelength required to maintain a constant peak to peak response amplitude. The resulting S(λ) had two peaks in all three rodent species, corresponding to ultraviolet and M cones, respectively: 359 nm and 511 nm for mice, 362 nm and 493 nm for gerbils, and 362 nm and 502 nm for rats. Results for mouse and gerbil were similar to literature reports of S(λ) functions obtained with other methods, confirming that the ERG associated to the AC Constant-Response Method was effective to obtain reliable S(λ) functions. In addition, due to its fast data collection time, the AC Constant Response Method has the advantage of keeping the eye in a constant light adapted state.     

Stimulation of the biosynthesis of carotenes by oxidative stress in Blakeslea trispora induced by elevated dissolved oxygen levels in the culture medium

The adaptive response of the fungus Blakeslea trispora to the oxidative stress induced by elevated dissolved oxygen concentrations during carotene production was investigated by measuring the specific activities of catalase (CAT) and superoxide dismutase (SOD) and the micromorphology of the fungus using a computerized image analysis system. Changes in the ratio of the volume of air (V_a) over the medium and the volume of medium (V_m) in the flask caused changes of the morphology of microorganism from clumps to pellets and increases in the specific activities of CAT and SOD. The oxidative stress in B. trispora resulted in a significant increase in carotene production, and a maximum proportion of β-carotene (60%), γ-carotene (50%), and lycopene (10%) (as percentages of total carotenes) was observed at a ratio V_a/V_m of 15.7, 4.0 and 1.5, respectively. The highest concentration of carotenes (115.0 mg/g dry biomass) was obtained in V_a/V_m ratio of 9.0.     

Quantification of dead ectomycorrhizae from forest soils using computerized image analysis

A first-generation regression model is presented, describing a linear relationship between dead ectomycorrhizal biomass (mg dry weight) and mycorrhizal area in the xy-plane (mm²) (r²=0.996). This model enables reliable estimates of dead ectomycorrhizal biomass in soils through area measurements of mycorrhizae using computerized image analysis.     

Steroids from Euphorbia and other latex-bearing plants

Steroids in latices isolated from 15 Euphorbia species and four other latex-bearing plants were analysed (in terms of their chemotaxonomy) using computerized GC-MS. Steroid patterns divided these plants into six groupings.     

Body shape and life style of the extinct rodent Canariomys bravoi (Mammalia,Murinae) from Tenerife,Canary Islands (Spain)

A three-dimensional reconstruction of the skeleton of the giant rat of Tenerife (Canary Islands, Spain) Canariomys bravoi was obtained by computerized microtomography. Body size, body mass, and body shape were estimated, and limb morphofunctional indices used to infer the style of life of this recently extinct rat. A sample of recent Murinae, including the Philippines endemic giant cloud rat Phloeomys cumingi, was used for comparison. It appears that C. bravoi differed from most continental rats by its relatively large size, body proportions, and tail length. Among its distinctive features, claws almost similarly developed on fore and hind limbs, and feet longer than hands evoke an intermediate body shape between rats and arboreal murines like Phloeomys. C. bravoi was a strong and powerfully muscled rat able to move on different substrates from floor to trees, and probably had digging skills.     

A pilot crossover study: effects of an intervention using an activity monitor with computerized game functions on physical activity and body composition

Background

Wearing an activity monitor as a motivational tool and incorporating a behavior-based reward system or a computerized game element might have a synergistic effect on an increase in daily physical activity, thereby inducing body fat reduction. This pilot crossover study aimed to examine the effects of a short-term lifestyle intervention using an activity monitor with computerized game functions on physical activity and body composition.

Methods

Twenty healthy volunteers (31 ± 3 years) participated in a 12-week crossover study. The participants were randomly assigned to either Group A (a 6-week game intervention followed by a 6-week normal intervention) or Group B (a 6-week normal intervention followed by a 6-week game intervention). The participants wore both a normal activity monitor (Lifecorder EX) and an activity monitor with computerized game functions (Yuuhokei) during the game intervention, whereas they only wore a normal activity monitor during the normal intervention. Before, during, and after the intervention, body composition was assessed.

Results

Significantly more daily steps were recorded for the game intervention than for the normal intervention (10,520 ± 562 versus 8,711 ± 523 steps/day, P < 0.01). The participants performed significantly more physical activity at an intensity of ≥ 3 metabolic equivalents (METs) in the game intervention than in the normal intervention (3.1 ± 0.2 versus 2.4 ± 0.2 METs · hour/day, P < 0.01). Although body mass and fat were significantly reduced in both periods (P < 0.01), the difference in body fat reduction was significantly greater in the game intervention than in the normal intervention (P < 0.05).

Conclusions

A short-term intervention using an activity monitor with computerized game functions increases physical activity and reduces body fat more effectively than an intervention using a standard activity monitor.     

L-carnosine as an enhancer to computerized cognitive behaviour therapy in Japanese workers,an unjustified claim

This letter comments on the conclusion drawn by Shirotsuki et al. (2017) in their article entitled “The effect for Japanese workers of a self-help computerized cognitive behaviour therapy program with a supplement soft drink”, recently published in BioPsychoSocial Medicine. The authors concluded that their drink, containing L-carnosine, enhances the effects of a computerized cognitive-behavioural therapy (CCBT) on the psychological well-being of healthy Japanese workers. Yet, we argue that their conclusion is unfounded given their results and the methodological shortcomings of their study. Briefly, while the authors reported improvement on the tension-anxiety subscale of the Profile of Mood States (POMS) in the CCBT only group, they also observed a lack of improvement on this subscale in the CCBT+L-carnosine group suggesting that the drink washes out this beneficial effect of CCBT. Methodological issues include the uncontrolled levels of L-carnosine metabolized by participants jeopardize the study’s internal validity. Also, the clinical meaningfulness of the findings seems dubious as post-treatment scores remained within the range of the general Japanese population. Consequently, we argue that Shirotsuki et al.’s study should be re-conducted before drawing any valid conclusion.     

A morphometric study of the midgut in resistant and non-resistant Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae) larvae to its nucleopolyhedrovirus (AgMNPV)

In this investigation, the anterior and posterior regions of the midgut of resistant (RL) and non-resistant (SL) Anticarsia gemmatalis larvae were analyzed morphometrically to characterize different regions along their length. Also, this investigation compares the results between SL and RL to improve the understanding of the resistance mechanisms to the virus. Histological sections were analyzed in a computerized system and the data were statistically analyzed by the Kruskal-Wallis test and by multivariate analysis. The midguts are morphometrically different in the two larval populations; we observed higher values in RL. The morphometric analysis of the epithelial cells showed that only columnar and goblet cells were distinct along the midgut, in both larvae, with the higher values found in the anterior region. Comparing the results between the two larval populations, all the epithelial cells presented significant differences, with RL showing the higher morphometric values. We concluded that there are regional differences along the length of midgut in SL and RL that confirm the idea of two morpho-functional distinct regions. The consistently morphometric superior values in RL indicate that this variability can be related with the resistance of A. gemmatalis to its AgMNPV.     

Are Normal Decision-Makers Sensitive to Changes in Value Contrast under Uncertainty? Evidence from the Iowa Gambling Task

The Iowa Gambling Task (IGT) developed by Bechara et al. in 1994 is used to diagnose patients with Ventromedial Medial Prefrontal Cortex (VMPFC) lesions, and it has become a landmark in research on decision making. According to Bechara et al., the manipulation of progressive increments of monetary value can normalize the performance of patients with VMPFC lesions; thus, they developed a computerized version of the IGT. However, the empirical results showed that patients'' performances did not improve as a result of this manipulation, which suggested that patients with VMPFC lesions performed myopically for future consequences. Using the original version of the IGT, some IGT studies have demonstrated that increments of monetary value significantly influence the performance of normal subjects in the IGT. However, other research has resulted in inconsistent findings. In this study, we used the computerized IGT (1X-IGT) and manipulated the value contrast of progressive increments (i.e., by designing the 10X-IGT, which contained 10 times of progressive increment) to investigate the influence of value contrast on the performance of normal subjects. The resulting empirical observations indicated that the value contrast (1X- vs. 10X-IGT) of the progressive increment had no effect on the performance of normal subjects. This study also provides a discussion of the issue of value in IGT-related studies. Moreover, we found the “prominent deck B phenomenon” in both versions of the IGT, which indicated that the normal subjects were guided mostly by the gain-loss frequency, rather than by the monetary value contrast. In sum, the behavioral performance of normal subjects demonstrated a low correlation with changes in monetary value, even in the 10X-IGT.     

Quantitative analysis of electroencephalograms: is there chaos in the future?

The history of quantitative, computerized electroencephalogram (EEG) analysis is reviewed. It is shown that, until very recently, the basic approach to EEG analysis involved the assumption that the EEG is stochastic. Consequently, statistical pattern recognition techniques, segmentation procedures, syntactic methods, knowledge-based approaches, and even artificial neural network methods have been developed with different levels of success. A fundamentally different approach to computerized EEG analysis, however, is making its way into the laboratories. The basic idea, inspired by recent advances in the area of non-linear dynamics, and especially the theory of chaos, is to view an EEG as the output of a deterministic system of relatively simple complexity, but containing non-linearities. This suggests that studying the geometrical dynamics of EEGs, and the development of neurophysiologically realistic models of EEG generation may produce more successful automated EEG analysis techniques than the classical, stochastic methods. Evidence supporting the non-linear dynamics paradigm is reviewed, and possible research paths are indicated.     

Regulatory properties of rabbit red blood cell hexokinase at conditions close to physiological

The true level of hexokinase in rabbit erythrocytes was determined by three different methods, including the spectrophotometric glucose-6-phosphate dehydrogenase coupled assay and a new radioisotopic assay. The value found at 37°C (pH 7.2) was 10.23±1.90 μmol/h per ml red blood cells, which is lower than previously reported values. More than 40 cellular components of the rabbit erythrocytes were tested for their effects on the enzyme. Their intracellular concentrations were also determined. Several of these compounds were found to be competitive inhibitors of the enzyme with respect to Mg·ATP^2?. Furthermore, reduced glutathione at a concentration of 1 mM was able to maintain hexokinase in the reduced state with full catalytic activity. The ability of orthophosphate to remove the inhibition of some phosphorylated compounds was examined under conditions similar to cellular (pH 7.2 and 50 μM of orthophosphate) and found to be of no practical interest. In contrast, the binding of ATP^4? and 2,3-diphosphoglycerate to the rabbit hemoglobin significantly modifies their intracellular concentrations and the formation of the respective Mg complexes. The pH-dependence of the reaction velocity and of the kinetic properties of the enzyme in different buffer systems were also considered. This information was computerized, and the rate of glucose phosphorylation in the presence of the mentioned compounds was determined. The value obtained, 1.94±0.02 μmol/h per ml red blood cells, is practically identical to the measured rate of glucose utilization by intact rabbit erythrocytes (1.92±0.3 μmol/h per ml red blood cells). These results provide further evidence for the central role of hexokinase in the regulation of red blood cell glycolysis.     

An Expanded Combined Evidence Approach to the Gavialis Problem Using Geometric Morphometric Data from Crocodylian Braincases and Eustachian Systems

The phylogenetic position of the Indian gharial (Gavialis gangeticus) is disputed - morphological characters place Gavialis as the sister to all other extant crocodylians, whereas molecular and combined analyses find Gavialis and the false gharial (Tomistoma schlegelii) to be sister taxa. Geometric morphometric techniques have only begun to be applied to this issue, but most of these studies have focused on the exterior of the skull. The braincase has provided useful phylogenetic information for basal crurotarsans, but has not been explored for the crown group. The Eustachian system is thought to vary phylogenetically in Crocodylia, but has not been analytically tested. To determine if gross morphology of the crocodylian braincase proves informative to the relationships of Gavialis and Tomistoma, we used two- and three-dimensional geometric morphometric approaches. Internal braincase images were obtained using high-resolution computerized tomography scans. A principal components analysis identified that the first component axis was primarily associated with size and did not show groupings that divide the specimens by phylogenetic affinity. Sliding semi-landmarks and a relative warp analysis indicate that a unique Eustachian morphology separates Gavialis from other extant members of Crocodylia. Ontogenetic expansion of the braincase results in a more dorsoventrally elongate median Eustachian canal. Changes in the shape of the Eustachian system do provide phylogenetic distinctions between major crocodylian clades. Each morphometric dataset, consisting of continuous morphological characters, was added independently to a combined cladistic analysis of discrete morphological and molecular characters. The braincase data alone produced a clade that included crocodylids and Gavialis, whereas the Eustachian data resulted in Gavialis being considered a basally divergent lineage. When each morphometric dataset was used in a combined analysis with discrete morphological and molecular characters, it generated a tree that matched the topology of the molecular phylogeny of Crocodylia.     

In vitro and in vivo D2-dopamine receptor binding with [123I]S(−)iodobenzamide ([123I]IBZM) in rat and human brain

As a promising dopamine D2-receptor imaging agent for single photon emission computerized tomography (SPECT), [¹²³I](S)-(−)-2-hydroxy-3-iodo-6-methoxy-N [(1-ethyl-2-pyrrolidinyl)methyl]benzamide ([¹²³I]IBZM) has recently been synthesized in a modified way along with its precursor, S(−)BZM, and the stereoisomer R(+)BZM. The present study applied this new product to investigate in vitro and in vivo D2-receptor binding in rat brain and in postmortem human brain. In vitro saturation binding curves with [¹²³I]IBZM for rat crude striatal membrane preparations yielded an affinity constant (K_d) of 0.28 nM confirming data in the literature. Displacement curves revealed an order of increasing potency as follows: R(+)BZM < S(−)sulpiride = < S(−)BZM < S(−)IBZM. A similar order was obtained when [³H]spiperone was used as ligand. For human putamen and caudate nucleus membranes slightly higher K_d values (0.49 nM) were obtained. Rank order of displacing potency for the various drugs was similar to that found in the rat preparations. In vivo uptake of [¹²³I]IBZM in rat brain following injection of 50 μCi (12–16pmol) in the tail vein revealed an increase in the striatum-to-cerebellum ratio from 1.5 at 5 min to 6.9 at 2 h. The olfactory tubercle-to-cerebellum ratio was also raised from 1.6 to 3.3. Other brain regions tested failed to show statistically significant enhancements. Coinjection of 40 nmol S(-)IBZM, 4μ mol S(−)BZM or 200 nmol haloperidol displaced [¹²³I]IBMZ when tested at 90 min. The use of 4μ mol R(+)BZM resulted in minor displacement only, demonstrating that stereospecificity of the displacement was present in vivo and in vitro. Displacements were also observed in substantia nigra and pons-medulla oblongata, but not in hippocampus or frontal and occipital cortex. The data provide the required background needed in order to initiate in vivo binding studies for D2-receptors in basal ganglia of human patients using [¹²³I]IBZM in SPECT analyses.     

Cluster Analysis of Longidorus Species (Nematoda: Longidoridae), a New Approach in Species Identification

Hierarchical cluster analysis based on female morphometric character means including body length, distance from vulva opening to anterior end, head width, odontostyle length, esophagus length, body width, tail length, and tail width were used to examine the morphometric relationships and create dendrograms for (i) 62 populations belonging to 9 Longidorus species from Arkansas, (ii) 137 published Longidorus species, and (iii) 137 published Longidorus species plus 86 populations of 16 Longidorus species from Arkansas and various other locations by using JMP 4.02 software (SAS Institute, Cary, NC). Cluster analysis dendograms visually illustrated the grouping and morphometric relationships of the species and populations. It provided a computerized statistical approach to assist by helping to identify and distinguish species, by indicating morphometric relationships among species, and by assisting with new species diagnosis. The preliminary species identification can be accomplished by running cluster analysis for unknown species together with the data matrix of known published Longidorus species.     

Specific effects of retinoic acid on the skeletal morphogenesis of the 11-day mouse embryo forelimb bud in vitro

Using our improved method for culturing 11-day mouse forelimb buds in vitro, we have investigated the effects of a local application of all-trans-retinoic acid (RA) on growth, cartilaginous differentiation and skeletal patterning in the mammalian limb bud. Carrier implants of catgut impregnated with DMSO or various doses of RA in DMSO were inserted at the apex of the buds in the proximo-distal axis just beneath the apical ectodermal ridge. After 6 days of culture, cartilaginous skeletons were stained and explants were processed for morphological analysis and quantitative study using computerized optical image analysis. Buds treated with low doses of RA exhibited stimulated growth and chondrogenesis. Moreover, hypertrophied and fused metacarpals were seen within explants treated with the lowest dose. High doses strongly inhibited growth and skeletal morphogenesis. An intermediate dose sustained cartilaginous differentiation at the same level as low doses, but concomitantly disturbed the skeletal pattern. These results are discussed considering reported RA effects on other experimental systems including avian limb bud as an in vivo model or cell cultures as an in vitro simplified model.