Maurocalcine is a novel toxin isolated from the venom of the chactid scorpion Scorpio maurus palmatus. It is a 33-mer basic peptide cross-linked by three disulfide bridges, which shares 82% sequence identity with imperatoxin A, a scorpion toxin from the venom of Pandinus imperator. Maurocalcine is peculiar in terms of structural properties since it does not possess any consensus motif reported so far in other scorpion toxins. Due to its low concentration in venom (0.5% of the proteins), maurocalcine was chemically synthesized by means of an optimized solid-phase method, and purified after folding/oxidation by using both C18 reversed-phase and ion exchange highpressure liquid chromatographies. The synthetic product (sMCa) was characterized. The half-cystine pairing pattern of sMCa was identified by enzyme-based cleavage and Edman sequencing. The pairings were Cys3-Cys17, Cys10-Cys21, and Cys16-Cys32. In vivo, the sMCa was lethal to mice following intracerebroventricular inoculation (LD 50 , 20 W Wg/mouse). In vitro, electrophysiological experiments based on recordings of single channels incorporated into planar lipid bilayers showed that sMCa potently and reversibly modifies channel gating behavior of the type 1 ryanodine receptor by inducing prominent subconductance behavior.z 2000 Federation of European Biochemical Societies.

Aims: To determine the distribution of the α1 to α6 chains of type IV collagen in Bruch's membrane of the human posterior pole. Methods: Cryosections (10 µm) from 18 human eyes (20 months to 83 years old) were acid treated, blocked with 10% normal goat serum, incubated for 1 hour with monoclonal antibodies against type IV collagen isoform specific peptides at 1:75 dilution, and visualised with an ABC staining kit. Results: In Bruch's membrane, the α1(IV) and α2(IV) chains were identified in retinal pigment epithelial (10/18 = 55%) and choriocapillaris basement membranes (18/18 = 100%); the α3(IV), α4(IV), and α5(IV) chains were also found in the retinal pigment epithelial basement membrane (13/18 = 72%). In the choroid, the α1(IV) and α2(IV) chains were detected in the blood vessels (18/18=100%). The α6(IV) chain was not identified in any sections. Conclusion:The heterogeneous distribution of α1-2(IV) and α3-5(IV) in Bruch's membrane could give insights into the function of this structure in health, ageing, and diseases such as age related macular degeneration.

Heart failure, a progressive, fatal disease of the heart muscle, is a state of chronic inflammation and injury. Heat shock protein (HSP) 72, a ubiquitous protective protein that is well-established as cardioprotective, is not increased in heart failure. In contrast, HSP60 levels are doubled in the failing heart. We hypothesized that HSF-1 is not activated in heart failure and that the increased expression of HSP60 was driven by NFκB activation. To test this hypothesis, we measured levels of heat shock factor (HSF) −1 and −2, the transcription factors controlling HSP expression, which were increased in heart failure. There was no increased phosphorylation of serine 230 or serine 303/307 in HSF-1, which are thought to regulate its activity; EMSA showed no increase in HSF binding activity with heart failure. Nonetheless, mRNA was increased for HSP60, but not HSP72. In contrast to HSF, NFκB activity was increased in heart failure. HSP60, but not HSP72, contained NFκB binding elements. ChIP assay demonstrated increased binding of NFκB to both of the NFκB binding elements in the heart failure HSP60 gene. TNFα treatment was used to test the role of NFκB activation in HSP60 expression in a cardiac cell line. TNFα increased HSP60 expression, and this could be prevented by pretreatment with siRNA inhibiting p65 expression. In conclusion, HSP72 is not increased in heart failure because HSF activity is not changed; increased expression of HSP60 may be driven by NFκB activation.

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Objectives The objective of this study was to compare the lesion detection performance of human observers between thin-section computed tomography images of the breast, with thick-section (>40 mm) simulated projection images of the breast. Methods Three radiologists and six physicists each executed a two alterative force choice (2AFC) study involving simulated spherical lesions placed mathematically into breast images produced on a prototype dedicated breast CT scanner. The breast image data sets from 88 patients were used to create 352 pairs of image data. Spherical lesions with diameters of 1, 2, 3, 5, and 11 mm were simulated and adaptively positioned into 3D breast CT image data sets; the native thin section (0.33 mm) images were averaged to produce images with different slice thicknesses; average section thicknesses of 0.33 mm, 0.71 mm, 1.5 mm, and 2.9 mm were representative of breast CT; the average 43 mm slice thickness served to simulate simulated projection images of the breast. Results The percent correct of the human observer’s responses were evaluated in the 2AFC experiments. Radiologists lesion detection performance was significantly (p<0.05) better in the case of thin-section images, compared to thick section images similar to mammography, for all but the 1 mm lesion diameter lesions. For example, the average of three radiologist’s performance for 3 mm diameter lesions was 92 % correct for thin section breast CT images while it was 67 % for the simulated projection images. A gradual reduction in observer performance was observed as the section thickness increased beyond about 1 mm. While a performance difference based on breast density was seen in both breast CT and the projection image results, the average radiologist performance using breast CT images in dense breasts outperformed the performance using simulated projection images in fatty breasts for all lesion diameters except 11 mm. The average radiologist performance outperformed that of the average physicist observer, however trends in performance were similar. Conclusions Human observers demonstrate significantly better mass-lesion detection performance on thin-section CT images of the breast, compared to thick-section simulated projection images of the breast, when the location of the lesion is known.