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Genomic and gene expression signature of the pre-invasive testicular carcinoma in situ.

Testicular cancer is the most common malignancy among men in the reproductive age and the incidence is increasing, probably caused by environmental factors. Most testicular cancers are testicular germ cell tumours and all originate from a carcinoma in situ (CIS) pattern. In this review, we focus on the pre-invasive CIS and its possible fetal origin by reviewing recent data originating from DNA microarrays and comparative genomic hybridisations. A comparison of gene expression and genomic aberrations reveal chromosomal "hot spots" with mutual clustering of gene expression and genomic amplification. Some of the genes found in the hot spots may be involved in creating the CIS phenotype. On the other hand, many genes that are highly expressed in CIS are not present in the hot-spot areas. The gene expression profile of CIS thus most likely reflects the combined result of genomic amplification and increased transcriptional activation and/or deficiency in the epigenetic silencing of specific loci. Amplification of chromosome 12p, appears to be a good genomic marker of the transition from the pre-malignant to malignant CIS cell; this is consistent with recent findings of propagation advantages in cultured undifferentiated embryonic stem cells after spontaneous amplification in similar regions. The gene expression profile of CIS cells has remarkable similarity to that of embryonic stem cells and supports our long-standing hypothesis of an early developmental origin of CIS and testicular germ cell cancer.

Carcinoma in Situ↗

Polymorphic growth in larvae of Maculinea butterflies, as an example of biennialism in myrmecophilous insects.

The presence of annual and biennial individuals within the same population has been recently demonstrated in the myrmecophilous butterflies Maculinea rebeli and Maculinea alcon, which present a cuckoo strategy inside Myrmica nests, and Maculinea arion which is a predatory species. Here, we present field and laboratory data on polymorphic larval growth in two other predatory species of Maculinea: M. teleius and M. nausithous. Body mass distributions of pre-pupation larvae were bimodal in both species. These results point to the existence of larvae that develop in 1 or 2 years. We also showed that the probability of pupation depended on larval body mass. In the case of M. teleius, the critical body mass at which larvae have a 50% probability of pupation is about 80 mg. We suggest that polymorphism in Maculinea may have evolved as an adaptation to life in ant nests, a habitat which protects them from predators and provides food. However, the quality of this resource is highly variable and unpredictable. According to the bet-hedging hypothesis, if the habitat is unpredictable, females should have an advantage by producing more variable offspring. In the case of Maculinea butterflies, this may involve maintaining larvae that develop in 1 or 2 years.

Animals↗

Beneficial in vitro effect of N-acetyl-cysteine on oxidative stress and apoptosis.

Chronic renal failure (CRF) is usually accompanied by abnormalities of both humoral and cellular immune response. The aim of the study was to investigate the influence of N-acetyl-cysteine (NAC) on intracellular oxidative stress and apoptosis rate of T lymphocytes in children with CRF. Twenty-two children (aged 4-16, mean 7.4) with CRF treated with dialysis were enrolled in the study. Intracellular reactive oxygen species (ROS) production was quantified by mean rhodamine 123 (RHO) fluorescence intensity with flow cytometry. Annexin V FITC was used for identifying apoptotic cells. Mean fluorescence intensity (MFI), which reflected intracellular oxidative stress in T lymphocytes, was increased in patients with CRF compared with the controls (CD3+: 31.58+/-11.58 vs 22.55+/-4.97, p = 0.043; CD3+CD4+: 32.50+/-8.59 vs 27.75+/-12.76, NS; CD3+CD8+: 32.10+/-11.85 vs 20.77+/- 4.89, p =0.012). Apoptotic T lymphocytes occurred more frequently in patients with CRF treated with hemodialysis (HD) (11.36+/-6.96%) than in the controls (6.14%+/-3.36%; p = 0.025). After 24 h incubation with NAC MFI and apoptosis rate decreased significantly in all subpopulations of lymphocytes. NAC, as a strong antioxidant, has a favorable effect on intracellular oxidative stress and apoptosis rate of T lymphocytes in patients with CRF. A decreased apoptosis rate may have positive effect on functional abnormalities of T cells already found in patients with CRF.

Acetylcysteine↗

Laparoscopic adrenalectomy of an adrenal adenoma with myelolipoma relieves severe hypertension in a 16-year-old patient.

Adrenal adenoma with myelolipoma is extremely rare in pediatrics. Although the tumor is usually asymptomatic, sometimes it may result in serious manifestations. A 16-year-old patient was diagnosed with severe hypertension associated with a right adrenal mass. The laboratory work-up was inconclusive of the nature of the tumor. Plasma and urinary hormonal studies were not diagnostic. Magnetic resonance imaging (MRI) of the brain and meta-iodobenzylguanidine (MIBG) scanning were normal. MRI of the abdomen showed a heterogeneous adrenal mass 4.2x3.3 cm. Laparoscopic resection of the mass was done, and the pathology revealed an adrenal adenoma with myelolipoma. After tumor resection the hypertension resolved, and within 1 month the patient was off medications. At 2-year follow-up the patient's blood pressure remained normal at 120/73 mmHg. This is a case in which an adrenal adenoma with myelolipoma, a benign and usually asymptomatic tumor, presented as severe hypertension resolving with surgical resection of the tumor.

Adolescent↗

Accumulation of poly(ADP-ribose) polymerase inhibitors in children with chronic renal failure.

Nicotinamide, N-methyl-2-pyridone-5-carboxamide (Met2PY) and N-methyl-4-pyridone-3-carboxamide (Met4PY) are biological metabolites of the intracellular coenzyme nicotinamide adenine dinucleotide (NAD) that can potentially inhibit poly(ADP-ribose) polymerase 1 (PARP-1; DNA repair enzyme). Our research was aimed at establishing whether chronic renal failure (CRF) in children leads to the elevation of plasma NAD metabolites sufficient to inhibit PARP-1 activity. Nicotinamide, Met2PY and Met4PY plasma and erythrocyte concentrations were measured in 25 children with CRF and in 19 healthy children. The effect of these NAD metabolites on PARP-1 activity was studied in vitro. We found that plasma concentration of all NAD metabolites (nicotinamide, Met2PY, Met4PY) in children with CRF could reach the concentration of 2, 30 and 10 microM as compared to 0.2, 1 and 0.5 microM, respectively, in healthy children. The concentration of nicotinamide metabolites correlated positively with plasma creatinine concentration and negatively with creatinine clearance in children with CRF. We found that Met2PY, Met4PY and nicotinamide inhibited in vitro PARP-1 activity with IC50 values of 2.1, 0.18 and 0.12 mM, respectively. Our data indicate that NAD metabolites accumulate in plasma of children with CRF and their combined effect could lead to the inhibition of PARP-1 activity. NAD metabolites could be particularly harmful in children due to higher DNA turnover than in adults.

Adolescent↗

Treatment for soft tissue sarcoma in childhood and adolescence. Austrian results within the CWS 96 study.

OBJECTIVE: The aim of the CWS 96 Study was to achieve an optimal treatment in children and adolescents with soft tissue sarcoma (STS) implementing a further refinement of risk-adapted allocation to chemotherapy, surgery and radiotherapy. METHODS: Treatment stratification was based on tumour histology, TNM status, postsurgical stage, localisation and age. Local tumour control was ensured by surgery and risk-adapted radiotherapy. RESULTS: From 1995 to 2002, 89 patients were registered in Austria. The 3-year event-free survival (EFS) and overall survival rates (OS) were 63% +/- 6% and 71% +/- 6%, respectively. 59/89 patients had localised RMS-like (rhabdomayosarcoma) STS (EFS 73% +/- 7%), 14 had localised NON-RMS STS (EFS 54% +/- 16%) and 15 patients had metastatic disease at diagnosis (EFS 33% +/- 12%), 1 patient had fibromatosis. The EFS rates at 3 years in patients with localised RMS-like tumours according to risk group were 92% +/- 8% for low and standard risk (12 patients) and 67% +/- 8% for high risk (47 patients). Favourable primary tumour sites of nonmetastatic RMS-like STS i.e. orbit, head/neck nonparameningeal or genitourinary non-bladder/prostate were diagnosed in 15 patients (1/15 patients died). In 44 patients with unfavourable localisation such as parameningeal, genitourinary bladder/prostate, extremity and others, 7 deceased. The 3 year EFS according to histology in patients with RMS-like STS was 61% +/- 11% for RME (embryonal RMS ) (28 patients) and 71% +/- 15% for RMA (alveolar RMS) (10 patients). The most common treatment failure was local relapse occurring in 21% of patients in the high-risk group. CONCLUSION: Risk-adapted individualisation of treatment led to a reduction of chemotherapy in the low and standard risk group without compromising survival. The outcome of RME and RMA was similar in this cohort of patients. These preliminary results after a median observation time of 2.5 years confirm the CWS 96 strategy.

Adolescent↗

What do patients living with advanced cancer and their carers want to know? - a needs assessment.

Our primary objective was to determine the content and format that is most suitable for educational events targeting patients and carers who are living with advanced cancer. Secondary objectives included examining the differences in information needs between patients and their carers, and providing an estimate of the rate of participation in educational events targeting such patients and carers. Out-patients receiving palliative radiotherapy at Toronto Sunnybrook Regional Cancer Center and their carers were invited to complete the Advanced Cancer Information Needs Survey. One hundred forty-four respondents participated in the survey. The participants identified the management of pain, fatigue, and home palliative care resources as the areas in which information was most needed. Carers displayed greater interest, and the range of topics in which they continue to seek additional information is wider. Thirty-one percent of respondents said they would participate in an educational event. A 'one-on-one' interview approach and short written materials were the preferred sources of information. Our study examined potential content areas and preferred format for proposed educational events targeting patients and carers living with advanced cancer. We also highlighted the reasons for, and potential limitations of this approach. The investment of future effort in evaluating the impact of 1 on 1 interviews and 'short written materials' on the informational needs of patients and carers living with advanced cancer is warranted.

Caregivers↗

Cell wall polysaccharides in differentiating anthers and pistils of Lolium perenne.

We are presenting the pattern of distribution of several carbohydrate epitopes, which constitute an important component of cell walls, within the anthers and pistils of a monocot grass species, perennial ryegrass (Lolium perenne L.). The results of immunocytochemical studies revealed that the flower organs are rich in (1-->3, 1-->4)-beta-D-glucans and possess surprisingly high amounts of methylesterified pectic domains that bind JIM7 antibody and pectin side chains rich in (1-->4)-beta-D-galactose residues which react with LM5 antibody. The presence of arabinogalactan protein epitopes binding JIM13 is restricted to microspores and ovule integuments. The results are discussed in terms of possible functions of cell wall polysaccharides and arabinogalactan proteins in the differentiation of flower organs.

Cell Differentiation↗

Properties of native and hydrophobic laccases immobilized in the liquid-crystalline cubic phase on electrodes.

Both native Trametes hirsuta laccase and the same laccase modified with palmytic chains to turn it more hydrophobic were prepared and studied with cyclic voltammetry and Raman spectroscopy. Native laccase immobilized in the monoolein cubic phase was characterized with resonance Raman spectroscopy, which demonstrated that the structure at the "blue" copper site of the protein remained intact. The diamond-type monoolein cubic phase prevents denaturation of enzymes on the electrode surface and provides contact of the enzyme with the electrode either directly or through the mediation by electroactive probes. Direct electron transfer for both laccases incorporated into a lyotropic liquid crystal was obtained under anaerobic conditions, whereas bioelectrocatalytic activity was shown only for the native enzyme. The differences in electrochemical behavior of native and hydrophobic laccase as well as possible mechanisms of direct and mediated electron transfers are discussed. The Michaelis constant for 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) diammonium salt (ABTS2-), KMapp, and the maximal current, Imax, for the native enzyme immobilized onto the electrode were estimated to be 0.24 mM, and 5.3 microA, respectively. The maximal current density and the efficiency of the catalysis, Imax/KMapp, were found to be 73 microA cm-2 and 208.2 microA cm-2 mM-1, respectively, and indicated a high efficiency of oxygen electroreduction by the enzyme in the presence of ABTS2- in the cubic-phase environment. Rate constants were calculated to be 7.5x10(4) and 3.6x10(4) M-1 s-1 for native and hydrophobic laccase, respectively.

Benzothiazoles↗

Remarks on surgical strategy in creating vascular access for hemodialysis: 18 years of one center's experience.

The aim of the study is to evaluate surgical methods for creating vascular access for hemodialysis (HD) in patients with chronic renal failure. Over the last 18 years, 1,827 surgical procedures were performed in 722 patients (399 men and 323 women, mean age 43.7 +/- 17 years) in order to provide and maintain permanent vascular access for HD. Among all the surgical procedures, 992 were based on the construction of arteriovenous fistulas (AVF) and 835 were undertaken as secondary reparative surgical procedures. A total of 992 vascular accesses have been performed, including 904 AVF on upper and 14 on lower extremities as well as insertion of 74 permanent catheters. Radiocephalic AVF (RCAVF) was the principal type of AVF (58.8%). While constructing secondary angio-access after using RCAVF on the other extremity, fistulas with usage of brachial vessels were preferred. A total of 228 AVF of this type were created, including 143 brachiocephalic (BCAVF) and 85 brachiobasilic (BBAVF) AVF. Lately, synthetic grafts (arteriovenous graft, AVG) have been used more frequently, in 90 AVF. A brachial straight graft was the main type procedure performed, with polytetrafluoroethylene (95.6%). The patency of the fistulas has been evaluated. Kaplan-Meier survival curves were calculated to determine primary, primary-assisted, and secondary patency. Log-rank analysis was used to determine differences between curves. Primary, primary-assisted, and secondary patency at 12 months and 24 months were calculated. Comparing AVF patency in two patients' age periods (18-65 years, >65 years), it may be concluded that in the elderly group AVG provides better treatment for AVF. Finally, we conclude that a multidisciplinary approach to vascular access strategy offers the best option to achieve good functional AVF. Autogenous arteriovenous access should be regarded as the most suitable type in creating VA. However, individual conditions should be taken into consideration.

Arteriovenous Shunt, Surgical↗

Cardiac valvular disease in patients with systemic lupus erythematosus. Relationship with anticardiolipin antibodies.

We performed two-dimensional and Doppler echocardiography in 52 patients with systemic lupus erythematosus and in 34 healthy controls. In 25 patients (48.1%) echocardiographic disturbances were found (25/52 vs 2/34, p<0.001). Valvular abnormalities were detected in 18 patients (34.6%) but in only two controls (18/52 vs 2/34, p<0.01). The mitral valve was involved in 12 patients (23.1%). The most frequent finding was mild (13.5%) and moderate (9.6%) regurgitation or valvular thickening (9.6%). The aortic valve was involved in six and the tricuspid valve in three patients (11.5% and 5.8%, respectively). Only one patient had echocardiographic non-infective verrucous vegetation affecting the tricuspid valve. We did not observe significant hemodynamic valve disease. Endocardial findings were related to disease duration (p<0.05) but not to disease activity. Twenty-eight SLE patients (53.8%) had increased anticardiolipin antibodies (aCL). Patients with aCL (particularly those with IgG class) were characterized by a high incidence of echocardiographic abnormalities (p<0.001), mainly valvular (mitral or aortic) regurgitation (p<0.05). We found a relationship between anticardiolipin antibodies and disease activity (p<0.05). In conclusion, we postulate a prominent role for anticardiolipin antibodies in the pathogenesis of heart valve disease in patients with SLE.

Adolescent↗

Peripheral blood lymphocyte apoptosis and circulating dendritic cells in patients with systemic lupus erythematosus: correlation with immunological status and disease-related symptoms.

We investigated in vivo the relationship between the degree of peripheral blood lymphocyte apoptosis and circulating dendritic cells (DC) as well as the immunological status in 45 patients with systemic lupus erythematous (SLE). Apoptosis was detected by phosphatidylserine externalization and assays to detect caspase activation. The total DC count (tDC) and their myeloid, mDC1 (BDCA1+) and mDC2 (BDCA3+), and plasmacytoid, pDC (BDCA3+), subtypes were assessed. Moreover, several immunological parameters, such as complement proteins, interferons (IFN), tumor necrosis factor (TNF)-alpha, interleukin (IL)-4, and IL-6 levels were assessed. There were no significant differences in both the intensity of apoptosis and DC counts between active and inactive SLE as well as between untreated patients and those treated with steroids. The incidence of lymphocyte apoptosis correlated positively with T-cell count, both T-helper (p=0.004) and cytotoxic T cells (p=0.001), but not with B or natural killer (NK) cells. The intensity of apoptosis enhanced along with the increase in complement C3 (p=0.016) and decrease in IFN-gamma (p=0.040) levels. The apoptotic cell count correlated with tDC (p=0.031), mDC1 (p=0.007), and pDC (p=0.039) counts. Both tDC and mDC1 counts correlated positively with antinuclear antibody (ANA) titers (p=0.017 and 0.032, respectively). Moreover, tDC correlated with C4 (p=0.039) and pDC with both C3 (p=0.032) and C4 (p=0.007) levels. The DC counts correlated inversely with IFN-gamma (tDC, p=0.038; mDC1, p=0.009), IL-6 (mDC2, p=0.031), or serum IgG levels (tDC, p=0.006; mDC1, p<0.001; mDC2, p=0.001). We found a positive correlation between lymphocyte apoptosis and peripheral blood DC count as well as the level of complement proteins in patients with SLE. The enhanced lymphocyte apoptosis was accompanied by the decrease in concentration of some cytokines, such as IFN-gamma or IL-6, as well as serum IgG level. This finding may reflect pathogenetic events during development of the disease, which include a persistent signal derived from circulating apoptotic lymphocytes, mobilizing the complement system, and attracting peripheral blood DC.

Adult↗

The changes in monosaccharide composition of immunoglobulin G in the course of rheumatoid arthritis.

The objective of this study was to determine whether galactosylation of immunoglobulin G (IgG) in patients with rheumatoid arthritis (RA) correlates with severity and duration of illness. Serum IgG glycosylation from 50 patients with RA in comparison with 30 healthy controls was analyzed. IgG from sera was isolated and monosaccharide composition was determined by means of gas chromatography. Ratio of galactose to mannose content was calculated. Patients were divided into groups according to three different criteria: disease duration, severity of RA (disease activity score index), and radiological degree of advancement of illness according to Steinbrocker. In patients with RA, significant decrease (p<0,01) of galactose ratio was observed in comparison with healthy control. In patients with long duration of RA (more than 15 years), significant decrease of galactose (p<0.05) ratio in comparison with patients who have had arthritis for less then 5 years was observed. For the group of patients with severe RA, we found reduction of galactose (p<0.001) ratio vs the group of patients in remission. For those patients who had radiological stage IV according to Steinbrocker, IgG galactose (p<0.01) content per oligosaccharide chain were also more decreased than in those patients who had stage I RA. Decreased galactosylation and of IgG in RA was observed. The lack of this carbohydrate component of IgG correlates with severity and duration of RA and could be used in monitoring the progression in early arthritis.

Adult↗

Project of Polish guidelines for conducting pharmacoeconomic evaluations in comparison to international health economic guidelines.

We present the project of Polish guidelines for conducting economic evaluation and compare them with international health economic guidelines, highlighting areas of agreement and dissent. The Polish guidelines are very similar to those in Australia, Canada, and some European Union countries. Nevertheless a number of implementation and attitude problems seem to have delayed the introduction of pharmacoeconomic evaluation as a prerequisite for reimbursement of new pharmaceutical products. A transitional phase is required for collecting reliable cost data and familiarizing and training personnel both in public authorities and private industry.

Journal Article↗

The anatomy of the chi-chi of Ginkgo biloba suggests a mode of elongation growth that is an alternative to growth driven by an apical meristem.

The chi-chi of Ginkgo biloba L. are cylindrical woody structures that grow downwards from the branches and trunks of old trees, eventually entering the soil where they give rise to adventitious shoots and roots. Examination of segments of young chi-chi taken from a mature ginkgo tree revealed an internal woody portion with irregular growth rings of tracheid-containing secondary xylem covered by a vascular cambium and bark. The cambium was composed of both fusiform cells and parenchymatous ray cells. Near the tip of the chi-chi, these two types of cambial cells had orientations ranging between axial, radial and circumferential with respect to the cylindrical form of the chi-chi. The xylem rays and tracheids that derived from the cambium showed correspondingly variable orientations. Towards the base of the chi-chi, the fusiform cells and young tracheids were aligned parallel to the axis, indicating that the orientation of the cambial cells in basal regions of the chi-chi gradually became normalised as the tip of the chi-chi extended forwards. Nevertheless, in such basal sites, tracheids near the centre of the chi-chi showed variable orientations in accordance with their mode of formation during the early stages of chi-chi development. The initiation of a chi-chi is proposed to derive from a localised hyperactivity of vascular cambial-cell production in the supporting stem. The chi-chi elongates by tip growth, but it does so in a manner different from organ growth driven by an apical meristem. It is suggested that the chi-chi of Ginkgo is an "evolutionary experiment" that makes use of the vascular cambium, not only for its widening growth but also for its elongation.

Ginkgo biloba↗

Computer-assisted bone age assessment: graphical user interface for image processing and comparison.

The current study is part of a project resulting in a computer-assisted analysis of a hand radiograph yielding an assessment of skeletal maturity. The image analysis is based on features selected from six regions of interest. At various stages of skeletal development different image processing problems have to be addressed. At the early stage, feature extraction is based on Lee filtering followed by the random Gibbs fields and mathematical morphology. Once the fusion starts, wavelet decomposition methods are implemented. The user interface displays the closest neighbors to each image under consideration. Results show the sensitivity of different regions to both stages of development and certain feature sensitivity within each region. At the early stage of development, the distal features are more reliable indicators, whereas at the stage of epiphyseal fusion, a larger dynamic range of middle features makes them more sensitive. In the current study, a graphical user interface has been designed and implemented for testing the image processing routines and comparing the results of quantitative image analysis with the visual interpretation of extracted regions of interest. The user interface may also serve as a teaching tool. At the later stage of the project it will be used as a classification tool.

Adolescent↗

Application of image analysis techniques in activated sludge wastewater treatment processes.

Image analytical techniques have been extensively developed to evaluate complex microbial aggregates such as sludge flocs and biofilms. This review covers the latest contributions concerning the application of image analysis to the activated sludge systems with respect to the most frequently used morphological parameters and relations between them and traditional wastewater treatment parameters. Recent developments have indicated that image analysis can be successfully used for the quantification of flocs and filamentous bacteria in the operating wastewater treatment plants, which enables prediction of bulking events and pinpoint flocs formation.

Bacteria↗