Validating a standardised test battery for synesthesia: Does the Synesthesia Battery reliably detect synesthesia?

Synesthesia is a neurological condition that gives rise to unusual secondary sensations (e.g., reading letters might trigger the experience of colour). Testing the consistency of these sensations over long time intervals is the behavioural gold standard assessment for detecting synesthesia (e.g., Simner, Mulvenna et al., 2006). In 2007 however, Eagleman and colleagues presented an online 'Synesthesia Battery' of tests aimed at identifying synesthesia by assessing consistency but within a single test session. This battery has been widely used but has never been previously validated against conventional long-term retesting, and with a randomly recruited sample from the general population. We recruited 2847 participants to complete The Synesthesia Battery and found the prevalence of grapheme-colour synesthesia in the general population to be 1.2%. This prevalence was in line with previous conventional prevalence estimates based on conventional long-term testing (e.g., Simner, Mulvenna et al., 2006). This reproduction of similar prevalence rates suggests that the Synesthesia Battery is indeed a valid methodology for assessing synesthesia.

Regulation and induction of CYP3A11, CYP3A13 and CYP3A25 in C57BL/6J mouse liver.

This study reports that dexamethasone (DEX) significantly induces CYP3A11, CYP3A13 and CYP3A25 mRNA expression in male and female 4 days, 3 weeks and 18 weeks old C57BL/6J mice. Furthermore, CYP3A activity, as measured by erythromycin-N-demethylation, is also significantly increased. PXR, RXRalpha and CAR are known to be involved in the induction of CYP3As. Here we report nuclear receptors PXR and RXRalpha but not CAR demonstrate gender- and age-dependent expression. Also, treatment of C57BL/6J mice with DEX induces PXR but not RXRalpha or CAR. In summary, we demonstrate DEX is not only able to up-regulate CYP3A expression and activity, but also the nuclear receptor PXR through which it may exert this effect. Furthermore, the gender- and age-dependent pattern of basal PXR and RXRalpha expression is similar to the 3 CYP3As analysed.

Expression of the Wilms' tumor suppressor gene, WT1, reduces the tumorigenicity of the leukemic cell line M1 in C.B-17 scid/scid mice.

The Wilms' tumor suppressor gene, WT1, encodes a transcription of the Cys2-His2 zinc finger type. Loss of WT1 gene function has been implicated in the development of malignancies including Wilms' tumor and acute leukemias. We have shown previously that ectopic expression of WT1 +KTS isoforms in murine M1 leukemic cells spontaneously induces monocytic differentiation without the requirement for external differentiation-inducing stimuli. To determine whether these observed effects in vitro corresponded to a reduction in tumorigenicity in vivo, parental M1, control M1.Neo, and M1.WT1 +KTS cells were transplanted into C.B-17 scid/scid mice, and the growth and metastatic behavior of the cell lines were monitored for a period of 20 weeks. Mice inoculated either s.c. on the flank or directly into the peritoneal cavity, with M1 cells stably expressing WT1 +KTS isoforms exhibited a marked decrease in tumor formation compared with control groups. Moreover, tumors arising in mice after the injection of M1.WT1 +KTS cells exhibited a loss in ectopic WT1 protein expression. Confirmation that the tumors arose from M1.WT1 +KTS cells was achieved by the amplification of the introduced transgene from tumor samples and indicates that the tumorigenicity of leukemic M1 cells in these animals correlates with a loss in WT1 expression. This investigation is the first to demonstrate the tumor-suppressive effects of WT1 expression in a leukemic cell line, further advancing the notion that WT1 acts as a differentiation-promoting gene during hematopoiesis and that loss of functional WT1 expression may contribute to leukemogenesis in vivo.

An HLA-A2 polyepitope vaccine for melanoma immunotherapy.

Epitope-based vaccination strategies designed to induce tumor-specific CD8 CTL are being widely considered for cancer immunotherapy. Here we describe a recombinant poxvirus vaccine that codes for ten HLA-A2-restricted epitopes derived from five melanoma Ags conjoined in an artificial polyepitope or polytope construct. Target cells infected with the melanoma polytope vaccinia were recognized by three different epitope-specific CTL lines derived from HLA-A2 melanoma patients, and CTL responses to seven of the epitopes were generated in at least one of six HLA-A2-transgenic mice immunized with the construct. CTL lines derived from vaccinated transgenic mice were also able to kill melanoma cells in vitro. Multiple epitopes within the polytope construct were therefore shown to be individually immunogenic, illustrating the feasibility of the polytope approach for melanoma immunotherapy. Tumor escape from CTL surveillance, through down regulation of individual tumor Ags and MHC alleles, might be overcome by polytope vaccines, which simultaneously target multiple cancer Ags.

Cloning and characterization of EphA3 (Hek) gene promoter: DNA methylation regulates expression in hematopoietic tumor cells.

The Eph family of receptor tyrosine kinases (RTK) has restricted temporal and spatial expression patterns during development, and several members are also found to be upregulated in tumors. Very little is known of the promoter elements or regulatory factors required for expression of Eph RTK genes. In this report we describe the identification and characterization of the EphA3 gene promoter region. A region of 86 bp located at -348 bp to -262 bp upstream from the transcription start site was identified as the basal promoter. This region was shown to be active in both EphA3-expressing and -nonexpressing cell lines, contrasting with the widely different levels of EphA3 expression. We noted a region rich in CpG dinucleotides downstream of the basal promoter. Using Southern blot analyses with methylation-sensitive restriction enzymes and bisulfite sequencing of genomic DNA, sites of DNA methylation were identified in hematopoietic cell lines which correlated with their levels of EphA3 gene expression. We showed that EphA3 was not methylated in normal tissues but that a subset of clinical samples from leukemia patients showed extensive methylation, similar to that observed in cell lines. These results suggest that DNA methylation may be an important mechanism regulating EphA3 transcription in hematopoietic tumors.

Tumor metastasis biopsy as a surrogate marker of response to melanoma immunotherapy.

In patients undergoing immunotherapy for metastatic melanoma, the clinical response in immunotherapeutic trials may be partial or difficult to detect. Tumor metastasis biopsy allows direct characterisation of an anti-tumor immunological response. During a phase I/II trial of granulocyte macrophage colony stimulating factor (GM-CSF) transduced autologous melanoma immunotherapy, the cellular response was examined by immunohistochemical analysis in a limited number of tumor biopsies taken from patients who either responded or progressed. Clinical response was associated with tumor infiltration by CD4+ and CD8+ T-cells, macrophages and differentiated dendritic cells (DC), and expression of HLA-DR by the tumor cells. This tumor infiltration was associated with increased melanoma-specific peripheral blood precursor cytotoxic T-lymphocyte (pCTL) and the ability to obtain tumor-infiltrating lymphocytes in vitro. In contrast, progression or a lack of clinical response was associated with a lack of T-cell and DC infiltration into the tumor tissue in all such biopsies. Macrophages and eosinophils infiltrated these tumors, while T-cells and DC were present at some distance from the tumor. These preliminary data strongly suggest that the location and extent of T-cell and DC infiltration, as well as the expression of HLA-DR by tumor cells are associated with a clinical response in this form of melanoma immunotherapy.

The anaesthesia consult clinic: does it matter which anaesthetist sees the patient?

PURPOSE: The objectives of this multicentre survey were: first to ascertain whether the preoperative evaluation performed by anaesthetists in the preadmission anaesthesia consultation clinic (PACC) is influenced by the knowledge that they will or will not be the patient's attending anaesthetist; and second to determine the agreement among anaesthetists with regard to investigations requested. METHODS: A postal survey was designed in two different versions, equal numbers of which were sent to 522 anaesthetists in 39 Canadian hospitals. The anaesthetists contacted were asked to consider how they would investigate two hypothetical patients in a PACC. One version of the survey stated that they would be the attending anaesthetist for the first patient, but not for the second patient (group A). In the second version the situation was reversed (group B). RESULTS: A total of 281 eligible replies were received. For each of the two patients the decision to order an echocardiogram, cardiac stress test, arterial blood gas analysis, pulmonary function tests, or internal medicine referral was not affected by the knowledge that the respondent would or would not be the patient's attending anaesthetist. Within each of the two groups there was very little consensus with regard to the ordering of laboratory tests. CONCLUSION: The extent of investigation in the PACC scenarios was not affected by knowledge of whether or not the consulting anaesthetist would be the attending anaesthetist in the operating room. However, there was minimal agreement among anaesthetists concerning the preoperative evaluation of the patients, regardless of who would be the anaesthetist on the day of operation. Efficiency in preoperative evaluation could be increased if anaesthetists saw their own patients in the PACC, or if clinical guidelines for patient assessment were introduced by departments.

Distribution of endogenous cobalamin between the transcobalamins in various mammals.

1. Plasma samples from ten mammals were chromatographed on Sephadex G-200 columns and the total cobalamin content of each fraction was determined. 2. Unlike the situation in man, the bulk of the endogenous plasma cobalamin was found attached to a transcobalamin II-like protein in all ten animals. Transcobalamin 0 carried between 3 and 20% and this proportion appeared to be inversely related to the plasma total cobalamin. No endogenous cobalamin peak corresponding to transcobalamin I was detected in any species, though in the rabbit 5.3% of the plasma total cobalamin was attached to a protein of apparent molecular weight 176 000.

A comparison of methods for analysing red cell and platelet volume-distribution curves.

Cell volume-distribution curves may be analysed using log-probability paper or by considering volumes corresponding to frequencies of 50 or 60% of that at the mode. All methods give similar results; the log-probability paper method is better because it allows the researcher to check that the underlying distribution is lognormal; methods based on 50 or 60% modal frequency are slightly less reliable but are simpler for routine use. For platelets it may be convenient to estimate the curve from the modal volume and the volume above the mode at which the frequency is reduced to 60% of the modal frequency.

Microcytosis, anisocytosis and the red cell indices in iron deficiency.

Red cell volume distribution curves have been used to measure microcytosis and anisocytosis in normal subjects, blood donors and patients with iron deficiency anaemia. These measurements were more sensitive than the conventional red cell indices for detecting blood donors with a low transferrin saturation. Three stages are suggested as iron deficiency progressively interferes with haemopoietic function. Anisocytosis and an increased percentage of microcytic cells are the first haematological abnormalities to occur and at this stage haemoglobin concentration is usually normal and trasferrin saturation less than 32%. At the second stage the MCV and MCH decline, haemoglobin concentration is generally sub-normal, though not below 9 g/dl, and transferrin saturation is usually below 16%. The final stage of iron deficiency is associated with a low MCHC, a haemoglobin concentration below 9 g/dl and a transferrin saturation of less than 16%.

The transport of endogenous vitamin B12 in normal human serum.

1. Serum from normal subjects has been chromatographed on Sephadex G-200 columns and the fractions containing transcobalamins 0, I and II have been identified. 2. The fractions corresponding to transcobalamin I contained, on average, 90% of the endogenous vitamin B12. Only 3% was attached to transcobalamin 0, and 7% was bound to transcobalamin II.

Measurement of the mean cell volume using electronic particle counters.

Under carefully controlled conditions electronic cell counters, for example the Coulter Counter, Model FN, and Channelyzer, may be calibrated to give MCV values down to as small as 20 fl which agree with those derived from the centrifugation PCV (corrected for plasma trapping) and the red cell count. The MCV values will be too high if the instrument uses a high cell concentration, has a fixed lower threshold, no effective upper threshold and no edit facility. This may partly explain why the Coulter Counter, Model S, when standardized with 4C cell control gives higher MCV values than the Model FN linked to the Channelyzer. The difference is, on average, 2 fl with normal blood samples and 5 fl in cases of microcytic anaemia. It is suggested that standards of low MCV should be used together with those of normal MCV when calibrating the Model S.

Determination of the packed cell volume using 131I-human serum albumin.

When trace quantities of labelled albumin have been added to blood, the packed cell volume (PCV) may be determined by isotope counting using the relationship: (plasma counts-blood counts)/(plasma counts). This method for measuring the PCV is not affected by plasma trapping between the red cells and the results do not depend upon the centrifugation conditions. The PCV determination with labelled albumin has a high precision (coefficient of variation = 0.9%) and there is very good agreement with the centrifugation PCV using Wintrobe tubes after this measurement has been corrected for plasma trapped between the red cells. No significant compression of the red cells occurred when the blood samples were centrifugated in Wintrobe PCV tubes.

Simple method for automating the differential leucocyte-count.

A simple method is described for analysing leucocyte-volume distribution curves to obtain a differential leucocyte-count. The results derived by this method agree well with the differential counts obtained by microscopical examination of the stained blood-film.