Determination of methotrexate and indomethacin in urine using SPE-LC-DAD after derivatization

A validated HPLC-DAD assay is presented for the simultaneous quantification of methotrexate and indomethacin in a drug combination which is used synergistically to intervene with tumoral or inflammatory tissue microenvironments. The analytes were isolated from urine via solid phase extraction. The method is based on derivatizing both analytes with a soluble carbodiimide coupler and 2-nitrophenylhydrazine directed to their commonly occurring carboxylate functions. The chromatographic separation was accomplished on an octylsilica column in less than 15 min using acetate buffer (pH 4; 10 mM)-methanol (60:40, v/v) as eluent at 1.5 ml/min and monitored at 400 nm. The selectivity of the method was demonstrated in a pre-dosed rheumatoid arthritis patient. Quality control samples were prepared and analyzed to reveal the validity of the method. Life samples collected from a healthy volunteer were monitored for both drugs and their urinary levels were determined.     

Haemocompatibility improvement of metallic surfaces by covalent immobilization of heparin-liposomes

Stainless steel surfaces were processed by means of plasma enhanced chemical vapor deposition (PE-CVD) fed with acrylic acid vapors in order to functionalize them with carboxyl groups, which were subsequently activated for covalent immobilization of heparin-loaded (HEP) NH(2) group-functionalized (Fun) nanoliposomes (NLs). Empty Fun or HEP non-functionalized (control) NLs were used as controls. NLs were characterized for mean diameter, surface charge and heparin encapsulation/release. Different lipid compositions were used for NL construction; PC/Chol (2:1mol/mol) or PC/Chol (4:1mol/mol) (fluid type vesicles) [which allow gradual release of heparin] and DSPC/Chol (2:1mol/mol) (rigid type vesicles). Surface haemocompatibility was tested by measuring blood clotting time. Platelet adhesion on surfaces was evaluated morphologically by SEM and CLSM. The haemocompatibility of plasma-processed surfaces was improved (compared to untreated surfaces); Fun-HEP NL-coated surfaces demonstrated highest coagulation times. For short surface/blood incubation periods, surfaces coated with Fun-HEP NLs consisting of PC/Chol (2:1) had higher coagulation times (compared to DSPC/Chol NLs) due to faster release of heparin. Heparin release rate from the various NL types and surface platelet adhesion results were in agreement with the corresponding blood coagulation times. Concluding, covalent immobilization of drug entrapping NLs on plasma processed surfaces is a potential method for preparation of controlled-rate drug-eluting metallic stents or devices.     

Embedding and Retrieving Private Metadata in Electrocardiograms

Due to the recent explosion of ‘identity theft’ cases, the safeguarding of private data has been the focus of many scientific efforts. Medical data contain a number of sensitive attributes, whose access the rightful owner would ideally like to disclose only to authorized personnel. One way of providing limited access to sensitive data is through means of encryption. In this work we follow a different path, by proposing the fusion of the sensitive metadata within the medical data. Our work is focused on medical time-series signals and in particular on Electrocardiograms (ECG). We present techniques that allow the embedding and retrieval of sensitive numerical data, such as the patient’s social security number or birth date, within the medical signal. The proposed technique not only allows the effective hiding of the sensitive metadata within the signal itself, but it additionally provides a way of authenticating the data ownership or providing assurances about the origin of the data. Our methodology builds upon watermarking notions, and presents the following desirable characteristics: (a) it does not distort important ECG characteristics, which are essential for proper medical diagnosis, (b) it allows not only the embedding but also the efficient retrieval of the embedded data, (c) it provides resilience and fault tolerance by employing multistage watermarks (both robust and fragile). Our experiments on real ECG data indicate the viability of the proposed scheme.

Role of mini-host models in the study of medically important fungi

Mini-host models have emerged as simple experimental systems to study the pathogenesis and host innate immune responses in fungal invaders and also to test drug efficacy against these organisms. A growing number of medically important fungi, including Aspergillus spp, Candida spp, Cryptococcus spp, and species in the class Zygomycetes, have been shown to infect and kill invertebrates such as roundworms, fruit flies, and wax moths. These studies have shown that several genes implicated in the virulence of fungi in mammalian models also have a similarly important pathogenic role in mini-host organisms. These mini-host models provide a unique opportunity of simultaneously exploring the molecular mechanisms of fungal pathogenicity and candidate agents with antifungal activity. Furthermore, the fact that some of these mini-hosts have well-defined genetics and conserved innate immunity offers the advantage of a comprehensive analysis of the molecular aspects of host immune response. We examine the relevance, advantages, and pitfalls of experimental systems of fungal infections in various mini-hosts and compare them with what is known in experimental systems in mammalian animal models.     

Myasthenia gravis accompanied by thymomas not related to foamy virus genome in Belarusian's patients

The spectrum and features of neurological disorders have been changed due to the Chernobyl catastrophe in the Republic of Belarus. More recently neurologists in Belarus have noted a significant increase in the frequency of myasthenia gravis (MG) with concomitant rise in the thymomas. There is some evidence suggesting that retroviruses play a key role in the development and pathogenesis of autoimmune diseases. This study analyzed thymomas from 45 MG patients from the Republic of Belarus by using PCR and primers for two regions of FV--gag and bel-2 genes. The results showed that none of the varied thymuses from the 45 MG patients contained FV genome. No relationship can be confirmed between FV and this disease and the results suggest that no pathological link between FV and MG exists.     

Association of endothelial shear stress with plaque thickness in a real three-dimensional left main coronary artery bifurcation model

Women with advanced heart failure have better prognosis than men (men versus women HR 2.5, 95% CI 1.1-5.5, p=0.03). Although interaction ischemic heart disease-gender was not significant, HR was 1.6 (95% CI 0.5-5.6, p=0.43) for patients with coronary artery disease and 3.4 (95% CI 1.1-10.5, p=0.03) for patients without.     

The role of glutamate and its receptors in migraine

Glutamate (Glu) is the principal excitatory neurotransmitter in the central nervous system. Its receptors are classified into ionotropic receptors, which are ion channels and include NMDA, AMPA and kainate receptors, named after the agonists that selectively bind to them, and metabotropic receptors, which are G-protein coupled receptors. The trigeminal system is considered to play a key role in migraine pathophysiology, trafficking pain signals from the head and face to the trigeminal nucleus caudalis. The role of glutamate in the pathophysiology of migraine is implicated by data from animal and human studies. Animal studies include experiments of cortical spreading depression, studies of c-fos protein expression in trigeminal nucleus caudalis, studies of plasma protein extravasation and electrophysiological studies. Human studies investigating the role of Glu in migraine pathogenesis measured the levels of Glu in plasma, platelets and cerebrospinal fluid, studied its effect on migraine symptoms and examined the effect of Glu in modulating sensitization. Findings from both the animal and the human studies suggest a link between glutamate and migraine and further suggest that glutamate plays a key role in migraine mechanisms. In the future, efforts should be made to further investigate the role of glutamate in migraine pathogenesis and, subsequently, in migraine treatment.     

Texture analysis of perimenopausal and post-menopausal endometrial tissue in grayscale transvaginal ultrasonography

The aim of this study was to investigate the feasibility of texture analysis in characterizing endometrial tissue as depicted in two-dimensional (2D) grayscale transvaginal ultrasonography. Digital transvaginal ultrasound endometrial images were acquired from 65 perimenopausal and post-menopausal women prior to gynaecological operations; histology revealed 15 malignant and 50 benign cases. Images were processed with a wavelet-based contrast enhancement technique. Three regions of interest (ROIs) were identified (endometrium, endometrium plus adjacent myometrium, layer containing endometrial-myometrial interface) on each original and processed image. 32 textural features were extracted from each ROI employing first and second order statistics texture analysis algorithms. Textural features-based models were generated for differentiating benign from malignant endometrial tissue using stepwise logistic regression analysis. Models' performance was evaluated by means of receiver operating characteristic (ROC) analysis. The best logistic regression model comprised seven textural features extracted from the ROIs determined on the processed images; three features were extracted from the endometrium, while four features were extracted from the layer containing the endometrial-myometrial interface. The area under the ROC curve (A(z)) was 0.956+/-0.038, providing 86.0% specificity at 93.3% sensitivity using the cut-off level of 0.5 for probability of malignancy. Texture analysis of 2D grayscale transvaginal ultrasound images can effectively differentiate malignant from benign endometrial tissue and may contribute to computer-aided diagnosis of endometrial cancer.     

Influence of attenuation correction by integrated low-dose CT on somatostatin receptor SPECT

AIM: Somatostatin receptor scintigraphy (SRS) is well-established in neuroendocrine tumour (NET) imaging. This study evaluated the impact of attenuation correction (AC) on SRS SPECT data in patients examined by SPECT-CT. METHODS: Planar scintigraphy and SPECT-CT of 17 patients (10 men, seven women; age, 40-74 years; mean, 62 years) suffering from NET were included. For the visual assessment of AC, the intensity and contrast of foci classified as pathological were rated in both the non-attenuation corrected (NAC) and the attenuation corrected (AC) SPECT images using a 5-point score. The change in signal intensity after AC was semiquantified two-fold for each focus in both SPECT(AC) and SPECT(NAC): firstly by using tumour-to-background (TB) ratios (defined as T(max)/B(mean)) for the determination of a TB(AC)/TB(NAC) ratio. Secondly, by a T(max,AC)/T(max,NAC) ratio. Both ratios were correlated to the focus depth. RESULTS: A total of 46 pathological foci were found. Focus contrast and intensity significantly increased in 14/46 foci (30%) after AC (mean, 3.7-4.0) in the visual analysis (P<0.001). While TB ratios increased only in 24/46 foci after AC and no correlation between the T(BAC)/T(BNAC) ratio and focus depth (r=0.027; P=0.856) was found, T(max) was higher after AC in all foci and the T(max,AC)/T(max,NAC) ratio showed the expected correlation to focus depth (r=0.650; P<0.01), indicating the superiority of the Tmax approach for the demonstration of the effects of attenuation correction on focal uptake. CONCLUSION: Attenuation correction of SRS SPECT data by SPECT-CT results in visually more clearly contrasted foci. Moreover, as focus intensity increases, especially in the more centrally localised foci, CT-based AC has a potential to further improve the sensitivity of SRS SPECT.