Bootstrap estimates of the statistical accuracy of thresholds obtained from psychometric functions

Estimates of the accuracy of a threshold obtained from a psychometric function are often based on asymptotic theory. When the number of trails is small, however, these estimates may be untrustworthy. A computer program is described that uses a move reliable bootstrap approach to obtaining estimates of the standard deviation and confidence limits of a threshold and of the slope and spread of the psychometric function for any criterion level of performance.     

Clonal Differences in Expression of 25-Hydroxyvitamin D3-1α-hydroxylase, of 25-Hydroxyvitamin D3-24-hydroxylase, and of the Vitamin D Receptor in Human Colon Carcinoma Cells: Effects of Epidermal Growth Factor and 1α,25-Dihydroxyvitamin D3

Human colon carcinoma cells express 25-hydroxyvitamin D₃-1α-hydroxylase (CYP27B1) and thus produce the vitamin D receptor (VDR) ligand 1α,25-dihydroxyvitamin D₃ (1,25-D3), which can be metabolized by 25-hydroxyvitamin D₃-24-hydroxylase (CYP24). Expression of VDR, CYP27B1, and CYP24 determines the efficacy of the antimitotic action of 1,25-D3 and is distinctly related to the degree of differentiation of cancerous lesions. In the present study we addressed the question of whether the effects of epidermal growth factor (EGF) and of 1,25-D3 on VDR, CYP27B1, and CYP24 gene expression in human colon carcinoma cell lines also depend on the degree of cellular differentiation. We were able to show that slowly dividing, highly differentiated Caco-2/15 cells responded in a dose-dependent manner to both EGF and 1,25-D3 by up-regulation of VDR and CYP27B1 expression, whereas in highly proliferative, less differentiated cell lines, such as Caco-2/AQ and COGA-1A and -1E, negative regulation was observed. CYP24 mRNA was inducible in all clones by 1,25-D3 but not by EGF. From the observed clonal differences in the regulatory effects of EGF and 1,25-D3 on VDR and CYP27B1 gene expression we suggest that VDR-mediated growth inhibition by 1,25-D3 would be efficient only in highly differentiated carcinomas even when under mitogenic stimulation by EGF.     

Haplotyping cryptic Adélie penguin taxa using low-cost,high-resolution melt curves

Distinguishing morphologically cryptic taxa, by definition, requires genetic data such as DNA sequences. However, DNA sequences may not be obtained easily for taxa from remote sites. Here we provide the details of a high-resolution melt-curve-based method using taxon-specific primers that can distinguish two taxa of Adélie penguins, and that will be usable in Antarctica when combined with some of the newly developed field-deployable thermal cyclers. We suggest that the wider adoption of field-deployable polymerase-chain-reaction-based techniques will enable faster assignation of haplotype to individuals in situ, and so allow the targeting of observations and sample collection to specimens relevant to the research question. Targeting individuals will also reduce the need to repeatedly handle animals and reduce the time and travel required to complete field work.     

A parametric study of freezing injury in AT-1 rat prostate tumor cells.

The connection between thermal history and cell injury in single AT-1 cells is studied systematically through a two-level, four-parameter (2(4)) experiment. The four parameters considered are cooling rate (CR), end temperature (ET), hold time (HT), and thawing rate (TR). Cryosurgically relevant high and low values of each parameter are chosen (CR, 5 to 50 degrees C/min; ET, -20 to -80 degrees C; HT, 0 to 15 min; TR, 20 to 200 degrees C/min) to maximize applicability of the results to cryosurgery; it is important to note that any conclusions drawn from the results are valid only for the range of parameter values studied. AT-1 cell suspensions are frozen in a controlled way on a directional solidification stage, and viability is assessed postthaw with a live/dead assay using the fluorescent dyes calcein-AM and propidium iodide to indicate live and dead cells, respectively. The parameters which most significantly affect short-term survival outcome are determined through calculation of the individual parameter effect values (E) according to the factorial experimental design guidelines. In addition, any synergy between two parameters in determining short-term survival outcome is revealed by calculation of the interaction value for those parameters (I). The results suggest that survival is most significantly affected by variation in end temperature and hold time, and the only significant parameter interaction found is between these two parameters. The analysis further suggests that survival depends nonlinearly on the thermal parameters, based on calculation of the survival curvature (C) in the parameter ranges studied. These results are discussed within the context of previously proposed mechanisms of cellular injury during freezing. Although coupling between several mechanisms is possible, single mechanisms which may explain the survival results include slow-cooling injury mechanisms such as solute effects injury, dehydration-induced membrane instabilities, and volume-catalyzed nucleation of intracellular ice.     

Single cell responses from the optic tectum of the zebra finch (Taeniopygia guttata castanotis Gould)

Responses of neurons of the optic tectum, the prominent, highly laminated mesencephalic station of the tectofugal visual pathway in birds, to computer-generated and other visual stimuli were examined in zebra finches. Our study shows that the contralateral retina projects to the tectum in topographic order. The representation of the visual field is tilted against the horizon by 22°. The representation of the contralateral hemifield extends to the ipsilateral side by 15°. Most neurons have receptive fields with excitatory centres of different shapes and inhibitory surround. A new type of neuronal receptive field is described which has an excitatory centre and a surround which is movement sensitive and preferably excited by very small spots. The first type of neurons is mostly located in upper tectal layers, the latter only in deeper layers. Excitatory centre sizes increase with depth, and there is a tendency of smaller receptive fields in the foveal region. The representation of the frontal visual field does not show specializations which could be expected if it were used for fixation of grain during pecking. Our results are in accordance with previous behavioural experiments. Accepted: 30 April 1999     

Accuracy of diagnostic registers and management of chronic obstructive pulmonary disease: the Devon primary care audit

Background

Guidelines on COPD diagnosis and management encourage primary care physicians to detect the disease at an early stage and to treat patients according to their condition and needs. Problems in guideline implementation include difficulties in diagnosis, using spirometry and the disputed role of reversibility testing. These lead to inaccurate diagnostic registers and inadequacy of administered treatments. This study represents an audit of COPD diagnosis and management in primary care practices in Devon.

Methods

Six hundred and thirty two patients on COPD registers in primary care practices were seen by a visiting Respiratory Specialist Nurse. Diagnoses were made according to the NICE guidelines. Reversibility testing was carried out either routinely or based on clinical indication in two sub-samples. Dyspnoea was assessed. Data were entered into a novel IT-based software which computed guideline-based treatment recommendations. Current and recommended treatments were compared.

Results

Five hundred and eighty patients had spirometry. Diagnoses of COPD were confirmed in 422 patients (73%). Thirty nine patients were identified as asthma only, 94 had normal spirometry, 23 were restrictive and 2 had a cardiac disorder. Reversibility testing changed diagnosis of 11% of patients with airflow obstruction, and severity grading in 18%. Three quarters of patients with COPD had been offered practical help with smoking cessation. Short and long-acting anticholinergics and long acting beta-2 agonists had been under-prescribed; in 15–18% of patients they were indicated but not received. Inhaled steroids had been over-prescribed (recommended in 17%; taken by 60%), whereas only 4% of patients with a chronic productive cough were receiving mucolytics. Pulmonary rehabilitation was not available in some areas and was under-used in other areas.

Conclusion

Diagnostic registers of COPD in primary care contain mistakes leading to inaccurate prevalence estimates and inappropriate treatment decisions. Use of pre-bronchodilator readings for diagnosis overestimates the prevalence and severity in a significant minority, thus post bronchodilator readings should be used. Management of stable COPD does often not correspond to guidelines. The IT system used in this study has the potential to improve diagnosis and management of COPD in primary care.     

Phylogenetic studies of the genus <Emphasis Type="Italic">Cebus</Emphasis>(Cebidae-Primates) using chromosome painting and G-banding

Background  

Chromosomal painting, using whole chromosome probes from humans and Saguinus oedipus, was used to establish karyotypic divergence among species of the genus Cebus, including C. olivaceus, C. albifrons, C. apella robustus and C. apella paraguayanus. Cytogenetic studies suggested that the species of this genus have conservative karyotypes, with diploid numbers ranging from 2n = 52 to 2n = 54.     

Activation of the unfolded protein response is required for defenses against bacterial pore-forming toxin in vivo

Pore-forming toxins (PFTs) constitute the single largest class of proteinaceous bacterial virulence factors and are made by many of the most important bacterial pathogens. Host responses to these toxins are complex and poorly understood. We find that the endoplasmic reticulum unfolded protein response (UPR) is activated upon exposure to PFTs both in Caenorhabditis elegans and in mammalian cells. Activation of the UPR is protective in vivo against PFTs since animals that lack either the ire-1-xbp-1 or the atf-6 arms of the UPR are more sensitive to PFT than wild-type animals. The UPR acts directly in the cells targeted by the PFT. Loss of the UPR leads to a normal response against unrelated toxins or a pathogenic bacterium, indicating its PFT-protective role is specific. The p38 mitogen-activated protein (MAPK) kinase pathway has been previously shown to be important for cellular defenses against PFTs. We find here that the UPR is one of the key downstream targets of the p38 MAPK pathway in response to PFT since loss of a functional p38 MAPK pathway leads to a failure of PFT to properly activate the ire-1-xbp-1 arm of the UPR. The UPR-mediated activation and response to PFTs is distinct from the canonical UPR-mediated response to unfolded proteins both in terms of its activation and functional sensitivities. These data demonstrate that the UPR, a fundamental intracellular pathway, can operate in intrinsic cellular defenses against bacterial attack.