Frequency analysis and clinical characterization of different types of spinocerebellar ataxia in Serbian patients.

The relative frequencies of different spinocerebellar ataxias (SCAs) vary widely among different ethnic groups, presumably due to a founder effect. We investigated the relative prevalence of SCA1-3, 6-8, 12, 17; dentate-rubro-pallidoluysian atrophy; and Friedreich's ataxia (FRDA) in Serbian patients with adult-onset (>20 years of age) hereditary and sporadic SCAs, and compared clinical features of patients with genetically confirmed SCAs. A total of 108 patients from 54 families (38 apparently dominant [ADCA] and 16 apparently recessive) with adult-onset hereditary ataxia and 75 apparently sporadic patients were assessed. Of 38 families with ADCA, 13 (34%) were positive for an expansion in an SCA1 and 5 families (13%) for an expansion in an SCA2 allele. In 20 families (53%), no expansions have been identified in any of the analyzed genes. Gaze palsy, spasticity, and hyperreflexia were significantly more common in SCA1, whereas slow saccades, hypotonia, hyporeflexia, and dystonia prevailed in SCA2 patients. Among the 16 families with an apparently recessive mode of ataxia inheritance, 4 (25%) were identified as having the FRDA mutation. Ataxia-causing mutations were identified in 8 (10.6%) of patients with apparently sporadic adult-onset ataxia.     

Multiple reference tissue method for contrast agent arterial input function estimation.

A precise contrast agent (CA) arterial input function (AIF) is important for accurate quantitative analysis of dynamic contrast-enhanced (DCE)-MRI. This paper proposes a method to estimate the AIF using the dynamic data from multiple reference tissues, assuming that their AIFs have the same shape, with a possible difference in bolus arrival time. By minimizing a cost function, one can simultaneously estimate the parameters and underlying AIF of the reference tissues. The method is computationally efficient and the estimated AIF is smooth and can have higher temporal resolution than the original data. Simulations suggest that this method can provide a reliable estimate of the AIF for DCE-MRI data with a moderate signal-to-noise ratio (SNR) and temporal resolution, and its performance increases significantly as the SNR and temporal resolution increase. As demonstrated by its clinical application, sufficient reference tissues can be easily obtained from normal tissues and subregions segmented from a tumor region of interest (ROI), which suggests this method can be generally applied to cancer-based DCE-MRI studies to estimate the AIF. This method is applicable to general kinetic models in DCE-MRI, as well as other CE imaging modalities.     

Optimal surgical treatment for bilateral multinodular goitre

Objective: In the present study, we compared subtotal thyroidectomy (STT) with total thyroidectomy (TT) in the management of bilateral multinodular goitre. Methods: A total of 204 consecutive patients with bilateral multinodular goitre were assigned to have either TT (n = 73) or STT (n = 131). Demographic details, hospital stay, biochemical findings, indications for operation and complications were noted. Results: There was no significant difference in the age and sex ratio between the two groups (P = 0.695 and P = 0.733). According to thyroid functional status, the majority of patients were euthyroid in both groups (73.28%vs 84.90%). Goitre grades II and III presented the most common indication for STT and TT. Hospital stay for patients who underwent TT was significantly longer compared to STT (P < 0.001). There was no significant difference in the rate of permanent complications. Conclusions: In the present study, we have shown that the risk of permanent complications with TT is no greater than with STT.     

Coexistence of Unverricht-Lundborg disease and congenital deafness: Molecular resolution of a complex comorbidity

Purpose :  We report on genetic analysis of a complex condition in a Serbian family of four siblings, wherein two had progressive myoclonic epilepsy (PME) and congenital deafness (CD), one had isolated congenital deafness (ICD), and one was healthy.
Methods and Results :  Molecular diagnosis performed by Southern blotting confirmed Unverricht-Lundborg disease in the available sibling with PME/CD. In the sibling with ICD (heterozygote for expansion mutation in CSTB ) we demonstrated recombination event between the D21S2040 marker and the CSTB gene and identified c.207delC (p.T70Xfs) mutation in the fourth exon of the transmembrane protease, serine-3 (TMPRSS3) gene (maps in close proximity to CSTB), responsible for nonsyndromic deafness in the sibling with PME/CD as well.
Discussion :  To the best of our knowledge this is the first genetic confirmation of the coexistence of these two mutations.     

A Large Sellar Granular Cell Tumor in a 21-year-old Woman

We report here the case of a 21-year-old woman with a large sellar tumor, extending to the suprasellar area associated with growth hormone deficiency, hypogonadism, hypocorticism, and hyperprolactinemia. Transsphenoidal surgery was performed, and histologic, immunohistochemical, and electron microscopic study lead to the diagnosis of granular cell tumor. These tumors are, in most cases, very small and are found incidentally at autopsy of older patients. Our case is exceptional because the tumor developed in a young woman, extended to the suprasellar region, and caused clinical symptoms.     

Synthetic oxygen carriers in cardiac tissue engineering

The prominence of cardiovascular diseases has prompted investigations into alternative treatment options, including tissue engineering. Currently, the biggest limitation in cardiac tissue engineering lies in delivering oxygen to all cells within the construct. Synthetic oxygen carriers hold much promise in that they have high affinity for oxygen and can be supplemented to culture medium without adverse effect on the cells. This review highlights two complementary studies by our group that utilized oxygen carriers in cardiac tissue engineering. Experimental and modeling studies were performed to evaluate the effect of a perfluorocarbon (PFC)-based synthetic oxygen carrier, Oxygent, on oxygen supply within tissue engineered cardiac constructs. Porous biorubber scaffolds with an array of parallel channels mimicking the capillary network were seeded with cardiomyocytes and fibroblasts, and cultivated in medium supplemented with PFC. The presence of PFC enhanced the transport of oxygen, increased oxygen concentrations, and yielded constructs that displayed stronger cardiac-like phenotype.     

Interactive effects of surface topography and pulsatile electrical field stimulation on orientation and elongation of fibroblasts and cardiomyocytes

In contractile tissues such as myocardium, functional properties are directly related to the cellular orientation and elongation. Thus, tissue engineering of functional cardiac patches critically depends on our understanding of the interaction between multiple guidance cues such as topographical, adhesive or electrical. The main objective of this study was to determine the interactive effects of contact guidance and electrical field stimulation on elongation and orientation of fibroblasts and cardiomyocytes, major cell populations of the myocardium. Polyvinyl surfaces were abraded using lapping paper with grain size 1-80 microm, resulting in V-shaped abrasions with the average abrasion peak-to-peak width in the range from 3 to 13 microm, and the average depth in the range from 140 to 700 nm (AFM). The surfaces with abrasions 13 microm wide and 700 nm deep, exhibited the strongest effect on neonatal rat cardiomyocyte elongation and orientation as well as statistically significant effect on orientation of fibroblasts, thus they were utilized for electrical field stimulation. Electrical field stimulation was performed using a regime of relevance for heart tissue in vivo as well as for cardiac tissue engineering. Stimulation (square pulses, 1 ms duration, 1 Hz, 2.3 or 4.6 V/cm) was initiated 24 h after cell seeding and maintained for additional 72 h. The cover slips were positioned between the carbon rod electrodes such that the abrasions were either parallel or perpendicular to the field lines. Non-abraded surfaces were utilized as controls. Field stimulation did not affect cell viability. The presence of a well-developed contractile apparatus in neonatal rat cardiomyocytes (staining for cardiac Troponin I and actin filaments) was identified in the groups cultivated on abraded surfaces in the presence of field stimulation. Overall we observed that (i) fibroblast and cardiomyocyte elongation on non-abraded surfaces was significantly enhanced by electrical field stimulation, (ii) electrical field stimulation promoted orientation of fibroblasts in the direction perpendicular to the field lines when the abrasions were also placed perpendicular to the field lines and (iii) topographical cues were a significantly stronger determinant of cardiomyocyte orientation than the electrical field stimulation. The orientation and elongation response of cardiomyocytes was completely abolished by inhibition of actin polymerization (Cytochalasin D) and only partially by inhibition of phosphatidyl-inositol 3 kinase (PI3K) pathway (LY294002).     

Obtaining patient-specific point model of the human ilium bone in the case of incomplete volumetric data using the method of parametric regions

In this paper, we present the methodology for determining the point model of the ilium bone in cases when volumetric data of the whole bone are not available. An extreme traumatic bone damage, osteoporosis, destruction of bone tissue by malignant bone tumors or the existence of only 2D medical image (X-ray) can be the reason for the lack of complete volumetric data. Points on the bone surface were defined at the curves that run through 26 previously defined parameters, at the edges of anteroposterior (A–P) and lateral projections and at the parts of the surface between some parameters. Those parts of the surface, enclosed by parameters, represent ten parametric regions. The values of coordinates, which represent the input data in the statistical program, were measured in a uniquely defined coordinate system. After establishing the correlations between the values of coordinates, 8869 different linear and nonlinear regression models were obtained. The prediction values for point coordinates were calculated and exported to a CAD program. Results obtained were tested on a randomly chosen male right ilium bone, applying the methodology for creating the prediction model using the method of parametric regions, which allows creating a complete polygonal model, for each region separately or just for some parts of the region. Results obtained in the form of regression equations for the right ilium bone can be applied to the left ilium bone. The results of the research were verified using a comparative deviation and distance analysis between the initial and obtained polygonal models.     

Immunoglobulins from sera of antiphospholipid syndrome patients are internalized in the HTR-8/SVneo cell line and cytotrophoblast in culture

Women with antiphospholipid syndrome (APS) experience pregnancy complications mostly due to impaired trophoblast cell functions. Antiphospholipid antibodies (aPL) affect extravillous trophoblast in vivo and in culture, but the mechanisms are still poorly understood. Previously, syncytiotrophoblast was shown to bind and internalize aPL, which was not replicated for extravillous cytotrophoblast in short term culture. Here, aPL binding and time dependent internalization was demonstrated with exposure to aPL in the extravillous cell line HTR-8/SVneo and isolated first trimester of pregnancy cytotrophoblast (CT) using immunocytochemistry and flow cytometry. Intracellular aPL were detectable from 2?h of culture, reaching 30.7?±?3.1% (p?<?0.001) positive cells in CT and 24.8?±?7% (p?<?0.01) in HTR-8/SVneo cells at 24?h and 33?±?4.2% (p?<?0.01) at 48?h. The data presented show that extravillous trophoblast cells internalize aPL in a time-dependent manner significantly more than control immunoglobulins after 24?h of exposure.