Immunodetection of retinoblastoma-related protein and its phosphorylated form in interphase and mitotic alfalfa cells

Plant retinoblastoma-related (RBR) proteins are primarily considered as key regulators of G(1)/S phase transition, with functional roles in a variety of cellular events during plant growth and organ development. Polyclonal antibody against the C-terminal region of the Arabidopsis RBR1 protein also specifically recognizes the alfalfa 115?kDa MsRBR protein, as shown by the antigen competition assay. The MsRBR protein was detected in all cell cycle phases, with a moderate increase in samples representing G(2)/M cells. Antibody against the human phospho-pRb peptide (Ser807/811) cross-reacted with the same 115?kDa MsRBR protein and with the in vitro phosphorylated MsRBR protein C-terminal fragment. Phospho-MsRBR protein was low in G(1) cells. Its amount increased upon entry into the S phase and remained high during the G(2)/M phases. Roscovitine treatment abolished the activity of alfalfa MsCDKA1;1 and MsCDKB2;1, and the phospho-MsRBR protein level was significantly decreased in the treated cells. Colchicine block increased the detected levels of both forms of MsRBR protein. Reduced levels of the MsRBR protein in cells at stationary phase or grown in hormone-free medium can be a sign of the division-dependent presence of plant RBR proteins. Immunolocalization of the phospho-MsRBR protein indicated spots of variable number and size in the labelled interphase nuclei and high signal intensity of nuclear granules in prophase. Structures similar to phospho-MsRBR proteins cannot be recognized in later mitotic phases. Based on the presented western blot and immunolocalization data, the possible involvement of RBR proteins in G(2)/M phase regulation in plant cells is discussed.     

Conformal stereotactic radiosurgery therapy: plan evaluation methods and results

The purpose of our study was the objective evaluation of micro-multileaf collimator (mMLC)-based stereotactic radiosurgery treatment plans. Forty-seven patients, 71 lesions received static beam conformal stereotactic radiosurgery treatment in our institute between November 2005 and June 2008. Target volume and organs at risk were outlined on a MRI-CT image fusion basis. BrainSCAN 5.31 system (BrainLAB AG, Heimstetten, Germany) was used for treatment planning, Elekta Presice TS linear accelerator (Elekta Oncology Systems Ltd, Crawley, UK) and BrainLAB m3 mMLC were used for treatment delivery. An invasive head frame, mounted to the treatment table, was used with four screws for patient head fixation. Treatment plans were analysed with objective parameters, such as conformal index (COIN), homogeneity index (HI), coverage index (CI) and healthy tissue relative overdose factor (HTOF) tools. x2 tests were performed between COIN, HI and the geometrical parameters of the target volume (lesion volume - LV, lesion-organ distance - LOD, lesion deformity index - LDI). Mean value of COIN, HI, HTOF and CI was 0.52 (SD 0.13), 1.16 (SD 0.1), 0.88 (SD 0.53), and 0.94 (SD 0.11), respectively. COIN significantly correlated with (p<0.001 in all three cases), while HI was independent of LV, LOD, LDI (p=0.94; 0.14 and 0.72). COIN is similar, HTOF is less than data from the literature. According to our results geometrical parameters of the target volume (size, location, deformation) significantly influence the COIN, but they have no effect on HI.     

Challenges in oncologic plastic surgery of the breast

Breast screening programs along with advances in diagnostic methods and oncologic treatment have resulted in full recovery for a decisive number of patients diagnosed with early-stage breast cancer. The results of the ultra-radical-, followed by the breast conserving era pose new opportunities and challenges for the oncologic breast surgeon. The focus of oncoplastic surgery is not only on the tumor, but also on the female patient, allowing for individualized immediate breast reconstruction with acceptable esthetic result following radical tumor exstirpation. Modern procedures differ both in concept and technique from that of traditional breast surgery. This paper provides a comprehensive and detailed overview of reconstructive and oncoplastic breast surgery.     

Vanilloid Receptor-1 (TRPV1) Expression and Function in the Vasculature of the Rat

Transient receptor potential (TRP) cation channels are emerging in vascular biology. In particular, the expression of the capsaicin receptor (TRPV1) was reported in vascular smooth muscle cells. This study characterized the arteriolar TRPV1 function and expression in the rat. TRPV1 mRNA was expressed in various vascular beds. Six commercially available antibodies were tested for TRPV1 specificity. Two of them were specific (immunostaining was abolished by blocking peptides) for neuronal TRPV1 and one recognized vascular TRPV1. TRPV1 was expressed in blood vessels in the skeletal muscle, mesenteric and skin tissues, as well as in the aorta and carotid arteries. TRPV1 expression was found to be regulated at the level of individual blood vessels, where some vessels expressed, while others did not express TRPV1 in the same tissue sections. Capsaicin (a TRPV1 agonist) evoked constrictions in skeletal muscle arteries and in the carotid artery, but had no effect on the femoral and mesenteric arteries or the aorta. In blood vessels, TRPV1 expression was detected in most of the large arteries, but there were striking differences at level of the small arteries. TRPV1 activity was suppressed in some isolated arteries. This tightly regulated expression and function suggests a physiological role for vascular TRPV1.     

Multimodality treatment resulting in long-term survival in hepatocellular carcinoma.

The early detection and complex therapy of the hepatocellular carcinoma (HCC) is one of the most seasonable questions of the gastroenterology-oncology, because of the increasing prevalence of the primary liver cancer. The course of the hepatocellular carcinoma is rapid, untreated patients rarely live over 5-6 months. Combination of different treatment modalities in HCC can offer the best chances for survival. If possible, a surgical resection should be the primary procedure, followed by adjuvant cytostatic treatment and chemoembolisation. The authors report three cases with HCC with extremely long survival. The long-term survival achieved by multimodality therapy, as presented in these cases, seems to justify aggressive therapeutical approaches in HCC. It has been concluded, that early detection and complex, aggressive multimodality treatment--even repeated liver resections and surgical elimination of duplex distant metastases--can result in long-term survival with a good quality of life.     

Natural and anthropogenic influences on the population structure of white‐tailed eagles in the Carpathian Basin and central Europe

European populations of the white‐tailed eagle Haliaeetus albicilla suffered a drastic decline during the 20th century. In many countries, only a few dozen breeding pairs survived or the species disappeared completely. By today, the populations have recovered, naturally or through restocking (e.g. in Scotland or the Czech Republic). In the Carpathian Basin, which is now a stronghold in southern Europe for the species in the southern part of the distribution range with more than 500 breeding pairs, only about 50 pairs survived the bottleneck. This region provides important wintering places for individuals arriving from different regions of Eurasia. In the present study, we investigated 249 DNA samples from several European countries, using 11 microsatellites and mitochondrial control region sequences (499 bp), to answer two main questions: 1) did the Carpathian Basin population recover through local population expansion or is there a significant gene flow from more distant populations as well? 2) Does the Czech population show signs in its genetic structure of the restocking with birds of unknown origin? Our microsatellite data yielded three genetically separate lineages within Europe: northern, central and southern, the latter being present exclusively in the Carpathian Basin. Sequencing of mitochondrial DNA revealed that there is one haplotype (B12) which is not only exclusive to the Carpathian Basin but it is frequent in this population. Our results suggest that in accordance with the presumably philopatric behaviour of the species, recovery of the Carpathian Basin population was mainly local, but some of the wintering birds coming from the northern and central populations contributed to its genetic composition as well. We detected considerably higher proportions of northern birds within the Czech Republic compared to the neighbouring areas, making it likely that parents of the reintroduced birds came from northern populations.     

Starch degradation by glucoamylase Glm from Saccharomycopsis fibuligera IFO 0111 in the presence and absence of a commercial pullulanase

This study describes the course of enzymatic hydrolysis of the native corn starches Maritena 100 and Maritena 300. Hydrolyses were carried out with glucoamylase Glm produced by Saccharomycopsis fibuligera IFO 0111, which degrades also native starch, with the purpose to substitute a two-step hydrolysis (amylase followed by glucoamylase) by a one-step process (glucoamylase only). Hydrolysis generally became more effective by adding the pullulanase Promozyme D, which cleaves alpha-1,6-glycosidic bonds more effectively than glucoamylase Glm does. The time course (kinetics) of hydrolysis was followed by determination of the glucose concentration and calculation of dextrose equivalents.     

Measuring Long-Term Impact Based on Network Centrality: Unraveling Cinematic Citations

Traditional measures of success for film, such as box-office revenue and critical acclaim, lack the ability to quantify long-lasting impact and depend on factors that are largely external to the craft itself. With the growing number of films that are being created and large-scale data becoming available through crowd-sourced online platforms, an endogenous measure of success that is not reliant on manual appraisal is of increasing importance. In this article we propose such a ranking method based on a combination of centrality indices. We apply the method to a network that contains several types of citations between more than 40,000 international feature films. From this network we derive a list of milestone films, which can be considered to constitute the foundations of cinema. In a comparison to various existing lists of ‘greatest’ films, such as personal favourite lists, voting lists, lists of individual experts, and lists deduced from expert polls, the selection of milestone films is more diverse in terms of genres, actors, and main creators. Our results shed light on the potential of a systematic quantitative investigation based on cinematic influences in identifying the most inspiring creations in world cinema. In a broader perspective, we introduce a novel research question to large-scale citation analysis, one of the most intriguing topics that have been at the forefront of scientific enquiries for the past fifty years and have led to the development of various network analytic methods. In doing so, we transfer widely studied approaches from citation analysis to the the newly emerging field of quantification efforts in the arts. The specific contribution of this paper consists in modelling the multidimensional cinematic references as a growing multiplex network and in developing a methodology for the identification of central films in this network.     

Pathophysiological significance of the global and regional left-ventricular ejection fraction (LV-EF) on ischaemic heart disease patients at rest and during ergometric exercise load (from radiocardio-cyclography (RKG-RCG) to parametric amplitude and phase scan)

As early as in the graphical RKG-RCG period a close inverse correlation could be established between the LV-EF and the serum myoglobin level during the acute course of myocardial infarction, in 10 patients in repeated follow up examinations. Corr. coeff. -0.91, p less than 0.01. In the mid seventies it could be shown by RKG-RCG, in 15 IHD patients with angina pectoris that the decrease of the basal LV-EF during ergometric load reflected the severity of IHD, compared with the increasing LV-EF tendency of 15 normal subjects. This fact could be verified on 19 middle age males (mean age, 41 years) by 99mTc RBC gamma camera ventriculography, i.e. that under modest load (100 W ergometry) a more than 10% decrease was a non-specific sign of main branch or three-vessel coronary heart disease. So in this extreme case our nuclear stethoscope-like RKG-RCG method alone may be satisfactory for staging and screening of coronary ischaemic heart disease (IHD) patients. All the 11 normal subjects belonged to the load-reaction group with more than 5% LV-EF increase, while the extensive anterior and inferior scar patients reacted without exception with more than 10% deficit (their basal LV-EF value was already under 45%). Supported by data in the literature in the comparison of load ECG and coronarography and two-step load, we could gain more refined data, but in accordance with the one-step load on the same patients. As regards the reproducibility of our global LV-EF investigations with gamma camera computer program Supersegams, it was within 5%.(ABSTRACT TRUNCATED AT 250 WORDS)