Preoperative CT for postoperative radiotherapy planning in breast cancer.

INTRODUCTION: The exact location of the original tumor should be known for a targeted increase in the dose to the tumor bed after breast cancer surgery. Therefore, at our site, we perform CT examinations of patients in the radiation position before breast cancer surgery. METHODS: Preoperative native CT scans were performed in the patients in the planning position for radiotherapy; these data were fused with standard planning CT for boost irradiation. We evaluated whether the tumor was accurately identifiable in preoperative CT scans. We also contoured one irradiation volume in the standard planning CT scans and the other in the fusion CT scans with preoperative examination, and compared these volumes. RESULTS: Out of the total number of 554 patients, we were able to identify the exact location of the breast tumor in 463 cases (83.6 %). In a group of 50 randomly selected patients, the clinical target volume for the boost dose to the postlumpectomy cavity was changed in 20 patients (40%) - decreased in 9 cases (18%) and increased in 11 cases (22%). CONCLUSION: As shown by the results of our study, preoperative CT in the planning position can be used in patients with confirmed breast cancer. This method allows us to more accurately locate the tumor bed and thus more accurately draw the target volume for boost irradiation. We confirmed that preoperative CT had an impact on the size of the target volume.

Long-Term Clinical Results of IGRT in Prostate Cancer Treatment.

BACKGROUND: The combination of intensity modulated radiation therapy (IMRT) and image guided radiotherapy (IGRT) plays a significant role in sparing normal tissue during prostate cancer treatment. We report the clinical outcomes of 260 patients treated with high-dose IGRT as well as the toxicity of high-dose IGRT in these patients. MATERIALS AND METHODS: From September 2008 to February 2012, 260 men with clinically localized prostate cancer underwent radical radiotherapy. Two hundred patients were treated with IMRT (78 Gy in 39 fractions) to the prostate and base of seminal vesicles using an adaptive protocol combining cone-beam computed tomography (CBCT) and kilovoltage image matching with individualized safety margin calculation. Sixty patients underwent treatment with the same prescribed dose using RapidArc with a reduced safety margin of 6 mm and daily online matching using CBCT. Late toxicity was scored prospectively according to the RTOG/FC-LENT scale. RESULTS: Eighteen patients (6.9%) experienced acute grade 2 gastrointestinal toxicity. There was no acute grade 3 or 4 gastrointestinal toxicity. Thirty-nine patients (15%) experienced acute grade 2 genitourinary toxicity and 6 patients (2.3%) had grade 3 gerourinary toxicity. Genitourinary toxicity grade 4 was observed in 5 (1.9%) patients, due to installation of a urinary catheter. At a median follow up of 84.2 months, the estimated 7-year cumulative incidences of grade 2 gastrointestinal and genitourinary toxicity were 4.4 and 7.1% respectively. The estimated 7-year prostate specific antigen relapse free survival was 97.1% for low-risk disease, 83.6% for intermediate-risk disease and 75% for high-risk patients. CONCLUSION: The use of IMRT in combination with IGRT results in a low rate of late toxicity. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 8. 9. 2019 Accepted: 25. 10. 2019.

Experience of stigma and discrimination in families of persons with schizophrenia in the Czech Republic.

RATIONALE: Mental health-related stigma and discrimination not only affect persons living with schizophrenia but also their whole families. Stigma and discrimination reduction is key to respond to the unmet needs of persons with mental illness. The local context is of particular importance in this endeavor, as stigma and its manifestations depend on the specific conditions of the target population and across cultures and settings. Evidence on effective approaches to reduce stigma is sparse and lacking from Central and Eastern Europe, including from the Czech Republic. OBJECTIVE: Our aim was to inform an anti-stigma campaign undertaken in the framework of the national mental health reform in the Czech Republic. METHODS: We conducted a qualitative study based on semi-structured in-depth interviews with relatives of patients diagnosed with schizophrenia in the Czech Republic. Initial respondents were identified through local mental health services and users' organizations with a consecutive chain-referral sampling. Transcribed narratives were thematically analyzed within a pre-developed four-level thematic framework to comprehensively identify experiences of stigma and discrimination in all areas of the respondents' lives. RESULTS: Stigma experiences of 25 diverse family members of persons living with schizophrenia spanned four levels of respondents' lives (macro-, meso-, micro-, and intro-level). The overarching issues were: (1) general lack of understanding and misconceptions about mental illness; (2) structural discrimination and paucity of governmental and public support system; (3) burden of "pervasive and unlimited" care and inability of independent living. CONCLUSIONS: We identified several features of mental health related stigma and the ensuing discrimination in Czech Republic experienced by persons with severe mental illness and their relatives. We developed a set of recommendations for policy-makers aimed at reducing ignorance and prejudice amongst the public and professionals, improving health and social services-including employment, housing and community integration-and the provision of family support.

Expression of inflammation-related genes in gluteal and abdominal subcutaneous adipose tissue during weight-reducing dietary intervention in obese women.

Accumulation of adipose tissue in lower body lowers risk of cardiovascular and metabolic disorders. The molecular basis of this protective effect of gluteofemoral depot is not clear. The aim of this study was to compare the profile of expression of inflammation-related genes in subcutaneous gluteal (sGAT) and abdominal (sAAT) adipose tissue at baseline and in response to multiphase weight-reducing dietary intervention (DI). 14 premenopausal healthy obese women underwent a 6 months' DI consisting of 1 month very-low-calorie-diet (VLCD), subsequent 2 months' low-calorie-diet and 3 months' weight maintenance diet (WM). Paired samples of sGAT and sAAT were obtained before and at the end of VLCD and WM periods. mRNA expression of 17 genes (macrophage markers, cytokines) was measured using RT-qPCR on chip-platform. At baseline, there were no differences in gene expression of macrophage markers and cytokines between sGAT and sAAT. The dynamic changes induced by DI were similar in both depots for all genes except for three cytokines (IL6, IL10, CCL2) that differed in their response during weight maintenance phase. The results show that, in obese women, there are no major differences between sGAT and sAAT in expression of inflammation-related genes at baseline conditions and in response to the weight-reducing DI.

Expression of lipolytic genes in adipose tissue is differentially regulated during multiple phases of dietary intervention in obese women.

The aim of this study was to investigate the time-course of the expression of key lipolysis-regulating genes in the subcutaneous adipose tissue (SCAT) during different phases of a 6-month dietary intervention. Fifteen obese women (BMI 34.7+/-1.0 kg.m(-2)) underwent a 6-month dietary intervention consisting of 1 month very low calorie diet (VLCD), followed by 2 months low calorie diet (LCD) and 3 months weight maintenance diet (WM). At each phase of the dietary intervention, a needle microbiopsy of the abdominal SCAT was obtained to evaluate mRNA expression of key lipolysis-regulating genes and a hyperinsulinemic euglycemic clamp (HEC) was performed. Dietary intervention induced a body weight reduction of 9.8 % and an improvement of insulin sensitivity as assessed by a HEC. Compared to pre-diet levels, mRNA levels of the adrenergic beta(2)-receptor in SCAT were higher at the end of VLCD and not different at the end of LCD and WM. In contrast, the expression of the adrenergic alpha(2)-receptor was lower at the end of VLCD and LCD compared to the pre-diet levels and did not differ at WM. Adipose triglyceride lipase and hormone-sensitive lipase levels were lower than the pre-diet levels at the end of LCD only, while phosphodiesterase-3B and the insulin receptor levels did not change throughout the dietary intervention. The results suggest that the regulation pattern of the genes that are involved in the control of lipolysis is different at the respective phases of the dietary intervention and depends on the duration of the diet and the status of energy balance.

Macrophage gene expression is related to obesity and the metabolic syndrome in human subcutaneous fat as well as in visceral fat.

AIMS/HYPOTHESIS: Our goal was to identify a set of human adipose tissue macrophage (ATM)-specific markers and investigate whether their gene expression in subcutaneous adipose tissue (SAT) as well as in visceral adipose tissue (VAT) is related to obesity and to the occurrence of the metabolic syndrome. METHODS: ATM-specific markers were identified by DNA microarray analysis of adipose tissue cell types isolated from SAT of lean and obese individuals. We then analysed gene expression of these markers by reverse transcription quantitative PCR in paired samples of SAT and VAT from 53 women stratified into four groups (lean, overweight, obese and obese with the metabolic syndrome). Anthropometric measurements, euglycaemic-hyperinsulinaemic clamp, blood analysis and computed tomography scans were performed. RESULTS: A panel of 24 genes was selected as ATM-specific markers based on overexpression in ATM compared with other adipose tissue cell types. In SAT and VAT, gene expression of ATM markers was lowest in lean and highest in the metabolic syndrome group. mRNA levels in the two fat depots were negatively correlated with glucose disposal rate and positively associated with indices of adiposity and the metabolic syndrome. CONCLUSIONS/INTERPRETATION: In humans, expression of ATM-specific genes increases with the degree of adiposity and correlates with markers of insulin resistance and the metabolic syndrome to a similar degree in SAT and in VAT.

Adiponectin inhibits spontaneous and catecholamine-induced lipolysis in human adipocytes of non-obese subjects through AMPK-dependent mechanisms.

Adiponectin is an adipokine increasing glucose and fatty acid metabolism and improving insulin sensitivity. The aim of this study was to investigate the role of adiponectin in the regulation of adipocyte lipolysis. Human adipocytes isolated from biopsies obtained during surgical operations from 16 non-obese and 17 obese subjects were incubated with 1) human adiponectin (20 microg/ml) or 2) 0.5 mM AICAR - activator of AMPK (adenosine monophosphate activated protein kinase). Following these incubations, isoprenaline was added (10(-6) M) to investigate the influence of adiponectin and AICAR on catecholamine-induced lipolysis. Glycerol concentration was measured as lipolysis marker. We observed that adiponectin suppressed spontaneous lipolysis by 21 % and isoprenaline-induced lipolysis by 14 % in non-obese subjects. These effects were not detectable in obese individuals, but statistically significant differences in the effect of adiponectin between obese and non-obese were not revealed by two way ANOVA test. The inhibitory effect of AICAR and adiponectin on lipolysis was reversed by Compound C. Our results suggest, that adiponectin in physiological concentrations inhibits spontaneous as well as catecholamine-induced lipolysis. This effect might be lower in obese individuals and this regulation seems to involve AMPK.

Dietary intervention-induced weight loss decreases macrophage content in adipose tissue of obese women.

OBJECTIVE: Accumulation of adipose tissue macrophages (ATMs) is observed in obesity and may participate in the development of insulin resistance and obesity-related complications. The aim of our study was to investigate the effect of long-term dietary intervention on ATM content in human adipose tissue. DESIGN: We performed a multi-phase longitudinal study. SUBJECTS AND MEASUREMENTS: A total of 27 obese pre-menopausal women (age 39 ± 2 years, body mass index 33.7 ± 0.5 kg m(-2)) underwent a 6-month dietary intervention consisting of two periods: 4 weeks of very low-calorie diet (VLCD) followed by weight stabilization composed of 2 months of low-calorie diet and 3 to 4 months of weight maintenance diet. At baseline and at the end of each dietary period, samples of subcutaneous adipose tissue (SAT) were obtained by needle biopsy and blood samples were drawn. ATMs were determined by flow cytometry using combinations of cell surface markers. Selected cytokine and chemokine plasma levels were measured using enzyme-linked immunosorbent assay. In addition, in a subgroup of 16 subjects, gene expression profiling of macrophage markers in SAT was performed using real-time PCR. RESULTS: Dietary intervention led to a significant decrease in body weight, plasma insulin and C-reactive protein levels. After VLCD, ATM content defined by CD45+/14+/206+ did not change, whereas it decreased at the end of the intervention. This decrease was associated with a downregulation of macrophage marker mRNA levels (CD14, CD163, CD68 and LYVE-1 (lymphatic vessel endothelial hyaluronan receptor-1)) and plasma levels of monocyte-chemoattractant protein-1 (MCP-1) and CXCL5 (chemokine (C-X-C motif) ligand 5). During the whole dietary intervention, the proportion of two ATM subpopulations distinguished by the CD16 marker was not changed. CONCLUSION: A 6-month weight-reducing dietary intervention, but not VLCD, promotes a decrease in the number of the whole ATM population with no change in the relative distribution of ATM subsets.

Sex chromosome evolution in cotton stainers of the genus Dysdercus (Heteroptera: Pyrrhocoridae).

The neo-X and neo-Y sex chromosomes of Dysdercus albofasciatus represent a unique model for the study of early stages of sex chromosome evolution since they retained the ability to pair and recombine, in contrast to sex chromosomes in most Heteroptera. Here we examined structure, molecular differentiation, and meiotic behaviour of the D. albofasciatus neo-sex chromosomes. Two related species with the ancestral X0 system, D. chaquensis and D. ruficollis, were used for a comparison. In D. albofasciatus, 2 nucleolar organizer regions (NORs) were identified on the neo-X chromosome using fluorescence in situ hybridization (FISH) with an rDNA probe, whereas a single NOR was found on an autosomal pair in the other 2 species. Genomic in situ hybridization (GISH) differentiated a part of the original X in the neo-X chromosome but not the neo-Y chromosome. The same segment of the neo-X chromosome was identified by Zoo-FISH with a chromosome painting probe derived from the X chromosome of D. ruficollis, indicating that this part is conserved between the species. Immunostaining against the cohesin subunit SMC3 revealed that only terminal regions of the D. albofasciatus neo-Xneo-Y bivalent pair and form a synaptonemal complex, which is in keeping with the occurrence of terminal chiasmata, whereas the interstitial region forms a large loop indicating the absence of homology. These results support the hypothesis that the neo-X chromosome evolved by insertion of the original X chromosome into 1 NOR-bearing autosome in an ancestor carrying the X0 system. As a consequence, the homologue of this NOR-autosome became the neo-Y chromosome. A subsequent inversion followed by transposition of the NOR located on the neo-Y onto the neo-X chromosome resulted in the present neo-sex chromosome system in D. albofasciatus.

[Relation between insulin resistance and amount of visceral adipose tissue]. / Vztah inzulíinové rezistence a mnozství viscerálního tuku.

BACKGROUND: Association of obesity with metabolic and cardiovascular complications depends on the adipose tissue distribution. The role of intraabdominal, i.e. visceral, adipose tissue in pathogenesis of insulin resistance is still not elucidated. The aim of this study was to investigate the relation between insulin resistance and contribution of visceral and subcutaneous fat in a group of women with a wide range of body weight. METHODS AND RESULTS: 62 women (age 21-66 years) among which 32 were non-obese and 30 obese (BMI > 30 kg/m2) were examined. The amount of visceral and subcutaneous fat was evaluated using computerized tomography, total body fat evaluated using bioimpedance, and the degree of insulin resistance was evaluated using glucose disposal (M) during euglycemic hyperinsulinemic clamp. Obese women had lower insulin sensitivity than non-obese (5.88 +/- 2.17 vs 3.32 +/- 1.44 mg/min/kg, p <0.001) and higher absolute amount of visceral fat. However, the relative amount of visceral fat (related to total body fat or subcutaneous fat) was not different between the two groups. In the entire study group, the magnitude of insulin sensitivity did correlate with absolute amount of total and visceral fat, but no correlation with relative amount of visceral fat was found. CONCLUSIONS: The results suggest that the absolute amount of fat, either total or visceral, is a stronger predictor of the degree of insulin resistance than the relative contribution of visceral fat.

Visfatin expression in subcutaneous adipose tissue of pre-menopausal women: relation to hormones and weight reduction.

BACKGROUND: A novel adipokine, visfatin, was found to be related to adiposity in humans and regulated by a number of hormonal signals. The aim of this study was to investigate the relationships of visfatin expression in adipose tissue with potential regulatory factors such as insulin, testosterone and tumor necrosis factor-alpha (TNF-alpha) and to elucidate the effect of a diet induced weight reduction on adipose tissue mRNA expression and plasma levels of visfatin. MATERIALS AND METHODS: Biopsies of subcutaneous abdominal adipose tissue (SCAAT) and plasma samples were obtained at the beginning of the study from 47 pre-menopausal women (age 38.7 +/- 1.7 years, body mass index (BMI) 27.9 +/- 1.4 kg m(-2)), consisting of 15 lean, 16 overweight and 16 obese subjects. The subgroup of 32 overweight/obese women (age 42.1 +/- 1.9 years, BMI 31.2 +/- 0.9 kg m(-2)) underwent a 12 week hypocaloric weight reducing diet and samples were obtained at the end of the diet. Biopsy samples were analysed for visfatin and TNF-alpha mRNA levels and plasma was analysed for relevant metabolites and hormones. RESULTS: In the group of 47 subjects visfatin mRNA expression in SCAAT was negatively correlated with plasma free testosterone (r = -0. 363, P < 0.05) and BMI (r = -0.558, P < 0.01) and positively associated with adipose tissue TNF-alpha mRNA expression (r = 0.688, P < 0.01). The diet resulted in the reduction of body weight and in the decrease of plasma insulin, free testosterone and TNF-alpha levels. In the group of overweight/obese subjects visfatin mRNA in SCAAT increased after the diet and the diet induced increase was positively correlated with the magnitude of body weight loss. CONCLUSION: Visfatin mRNA expression in SCAAT is associated with TNF-alpha expression, plasma free testosterone and BMI in pre-menopausal women. A weight reducing hypocaloric diet results in the increase of visfatin mRNA in SCAAT.

Retinol-binding protein 4 expression in visceral and subcutaneous fat in human obesity.

Retinol binding protein 4 (RBP4) is a novel adipokine which might be involved in the development of insulin resistance. The aim of the study was to investigate the expression of RBP4 mRNA in subcutaneous and visceral fat depots and the relationship between RBP4 plasma and mRNA levels relative to indices of adiposity and insulin resistance. In 59 Caucasian women (BMI 20 to 49 kg/m(2)) paired samples of subcutaneous and visceral fat were obtained for RBP4, leptin and GLUT 4 mRNA analysis using reverse transcription-quantitative PCR. Euglycemic hyperinsulinemic clamp and computed tomography scans were performed. RBP4 mRNA levels as well as GLUT 4 mRNA and leptin mRNA levels were lower (P<0.001, P<0.01 and P<0.001, respectively) in visceral compared to subcutaneous fat. No differences were found in RBP4 mRNA expression in the two fat depots or in RBP4 plasma levels between subgroups of non-obese subjects (n=26), obese subjects without metabolic syndrome (n=17) and with metabolic syndrome (n=16). No correlations between RBP4 mRNA or plasma levels relative to adiposity, glucose disposal rate and GLUT 4 mRNA expression in adipose tissue were found. There was a weak positive correlation between plasma RBP4 and plasma triglycerides (r = 0.30, p<0.05) and between plasma RBP4 and blood glucose (r = 0.26, p<0.05). Regardless of the state of adiposity or insulin resistance, RBP4 expression in humans was lower in visceral than in subcutaneous fat. We found no direct relationship between either RBP4 mRNA or its plasma levels and the adiposity or insulin resistance.

The atrial natriuretic peptide- and catecholamine-induced lipolysis and expression of related genes in adipose tissue in hypothyroid and hyperthyroid patients.

Thyroid dysfunction is associated with several abnormalities in intermediary metabolism, including impairment of lipolytic response to catecholamines in subcutaneous abdominal adipose tissue (SCAAT). Atrial natriuretic peptide (ANP) is a powerful lipolytic peptide; however, the role of ANP-mediated lipolysis in thyroid disease has not been elucidated. The aim of this study was to investigate the role of thyroid hormones in the regulation of ANP-induced lipolysis as well as in the gene expression of hormone-sensitive lipase, phosphodiesterase 3B (PDE3B), uncoupling protein-2 (UCP2), natriuretic peptide receptor type A, and beta(2)-adrenergic receptor in SCAAT of hyperthyroid and hypothyroid patients. Gene expression in SCAAT was studied in 13 hypothyroid and 11 hyperthyroid age-matched women before and 2-4 mo after the normalization of their thyroid status. A microdialysis study was performed on a subset of nine hyperthyroid and 10 hypothyroid subjects. ANP- and isoprenaline-induced lipolyses were higher in hyperthyroid subjects, with no differences between the groups following treatment. Hormone-sensitive lipase gene expression was higher in hyperthyroid compared with hypothyroid subjects before treatment, whereas no difference was observed following treatment. No differences in gene expression of other genes were observed between the two groups. Following treatment, the gene expression of UCP2 decreased in hyperthyroid, whereas the expression of PDE3B decreased in hypothyroid subjects. We conclude that thyroid hormones regulate ANP- and isoprenaline-mediated lipolysis in human SCAAT in vivo. Increased lipolytic subcutaneous adipose tissue response in hyperthyroid patients may involve postreceptor signaling mechanisms.

Isoflavonoids are present in Arabidopsis thaliana despite the absence of any homologue to known isoflavonoid synthases.

Extracts from Arabidopsis thaliana leaves and inflorescence stalks and from Lepidium sativa (Brassicaceae) seedlings were analysed by HPLC-MS-SIM and by five isoflavonoid-specific ELISA methods after the HPLC fractionation of samples, in order to determine presence of isoflavonoids. Individual ELISAs were specific for daidzein, genistein, biochanin A and for their derivatives substituted either at the 4'- or at the 7- positions. Both analytical approaches revealed homologous spectra of isoflavonoids in both plant species. As the ononin specific immunoassay was not available this compound was only detected by HPLC-MS. Formononetin and prunetin represented the main aglycones followed by biochanin A, daidzein and genistein; sissotrin was the most abundant isoflavonoid glycoside followed by ononin, daidzin and genistin. The content of individual compounds ranged from a few micrograms up to 2.2 mg kg(-1) (dry weight). Expression profiles of A. thaliana genes homologous to enzymes involved in isoflavonoid synthesis and metabolism were extracted from publicly available transcriptomic datasets for various tissues. Genes likely to be involved in important steps of the isoflavonoid metabolism in A. thaliana were identified. However, in accord with the previously published data, no homologue to isoflavone synthases known from the Fabaceae plants was found. These aryl migrating enzymes belong to the CYP93C family that is absent in A. thaliana. We conclude that another gene must be responsible for biosynthesis of the isoflavone skeleton in Brassicaceae.

[Carbomers and their use in pharmaceutical technology]. / Karbomery a jejich vyuzití ve farmaceutické technologii.

Carbomers, high-molecular polymers based on acrylic acid, are employed in the technology of drugs as auxiliary substances acting as emulsifiers, gel-forming substances, stabilizers of suspensions, and binders in tablets. Their properties are utilized to control release as well as to improve biological availability of drugs. They are physiologically inert, non-sensitizing, and possess excellent thermal stability. They are used for various routes of administration: topical, oral and peroral, vaginal and rectal.

The use of some hydrophobic substances in tablet technology.

The objective of this work is to continue in a study of utilization of synthetic triacylglycerides of higher aliphatic acids as additives improving the flow properties of granulates and reducing die friction in the tabletting press. These effects are not supposed to interfere with the drug liberation velocity. We have evaluated the effects of the synthetic triacylglycerides in theophylline containing model tablets using magnesium stearate as the reference standard. Magnesium stearate is the lubricant of choice in the production, found in up to 80% of all industrially produced tablets.

[Drug forms for drug delivery into the gastrointestinal tract]. / Liekové formy na prívod lieciva do gastrointestinálneho traktu.

Most drugs introduced into the GIT are absorbed into the stomach and/or the small intestine according to their chemical character. Some drugs (their number is on the increase) require targeted supply to a certain site in the GIT if their biological activity is to be manifested and made use of. Fulfilling this condition, which concerns primarily peptides and proteins, requires adjustments of dosage forms. Much effort is put forth to shift absorption into the large intestine (colon), which is advantageous from the viewpoint of circadian biorhythms.