Chromosomal in situ suppression hybridization of human gonosomes and autosomes and its use in clinical cytogenetics

DNA libraries from sorted human gonosomes were used selectively to stain the X and Y chromosomes in normal and aberrant cultured human cells by chromosomal in situ suppression (CISS-) hybridization. The entire X chromosome was stained in metaphase spreads. Interphase chromosome domains of both the active and inactive X were clearly delineated. CISS-hybridization of the Y chromosome resulted in the specific decoration of the euchromatic part (Ypter-q11), whereas the heterochromatic part (Yq12) remained unlabeled. The stained part of the Y chromosome formed a compact domain in interphase nuclei. This approach was applied to amniotic fluid cells containing a ring chromosome of unknown origin (47,XY; +r). The ring chromosome was not stained by library probes from the gonosomes, thereby suggesting its autosomal origin. The sensitivity of CISS-hybridization was demonstrated by the detection of small translocations and fragments in human lymphocyte metaphase spreads after irradiation with 60Co-gamma-rays. Lymphocyte cultures from two XX-males were investigated by CISS-hybridization with Y-library probes. In both cases, metaphase spreads demonstrated a translocation of Yp-material to the short arm of an X chromosome. The translocated Y-material could also be demonstrated directly in interphase nuclei. CISS-hybridization of autosomes 7 and 13 was used for prenatal diagnosis in a case with a known balanced translocation t(7;13) in the father. The same translocation was observed in amniotic fluid cells from the fetus. Specific staining of the chromosomes involved in such translocations will be particularly important, in the future, in cases that cannot be solved reliably by conventional chromosome banding alone

Preparation of micron and submicron particles via spray drying and electrostatic precipitation

Small particles are of great interest in a variety of applications. Spray drying is a common technique for particle synthesis, but it is limited with respect to sizes below 10 μm. Therefore, a new laboratory-scale spray dryer was designed to address this issue. A novel aerosol generator consisting of a piezo crystal in a swirl chamber was designed to obtain droplets in the low micrometer range. After drying and cooling, particles were deposited in a molten carrier, using melt electrostatic precipitation. The median particle size of three pharmaceutical drug substances was 2 μm. The size distribution was particularly narrow, with span values of about 1.1. Spray drying is a sufficient technique to produce small drug particles below 5 μm with ultrasonic atomization at high frequencies. Electrostatic precipitation in a molten carrier is a suitable method to capture these particles

Applying a correction procedure to the prevalence estimates of overweight and obesity in the German part of the HBSC study

Background: Prevalence rates for overweight and obesity based on self-reported height and weight are underestimated, whereas the prevalence rate for underweight is slightly overestimated. Therefore a correction is needed. Aim of this study is to apply correction procedures to the prevalence rates developed on basis of (self-reported and measured) data from the representative German National Health Interview and Examination Survey for Children and Adolescents (KiGGS) to (self-reported) data from the German Health Behaviour in School Aged Children (HBSC) study to determine whether correction leads to higher prevalence estimates of overweight and obesity as well as lower prevalence rates for underweight. Methods: BMI classifications based on self-reported and measured height and weight from a subsample of the KiGGS study (2,565 adolescents aged 11–15) were used to estimate two different correction formulas. The first and the second correction function are described. Furthermore, the both formulas were applied to the prevalence rates from the HBSC study (7,274 adolescents aged 11–15) which are based on self-reports collected via self-administered questionnaires. Results: After applying the first correction function to self-reported data of the HBSC study, the prevalence rates of overweight and obesity increased from 5.5% to 7.8% (compared to 10.4% in the KiGGS study) and 2.7% to 3.8% (compared to 7.8% in the KiGGS study), respectively, whereas the corrected prevalence rates of underweight and severe underweight decreased from 8.0% to 6.7% (compared to 5.7% in the KiGGS study) and from 5.5% to 3.3% (compared to 2.4% in the KiGGS study), respectively. Application of the second correction function, which additionally considers body image, led to further slight corrections with an increase of the prevalence rates for overweight to 7.9% and for obese to 3.9%. Conclusion: Subjective BMI can be used to determine the prevalence of overweight and obesity among children and adolescents. Where there is evidence of bias, the prevalence estimates should be corrected using conditional probabilities that link measured and subjectively assessed BMI from a representative validation study. These corrections may be improved further by considering body image as an additional influential factor

Discovery of the cancer stem cell related determinants of radioresistance

AbstractTumors are known to be heterogeneous containing a dynamic mixture of phenotypically and functionally different tumor cells. The two concepts attempting to explain the origin of intratumor heterogeneity are the cancer stem cell hypothesis and the clonal evolution model. The stochastic model argues that tumors are biologically homogenous and all cancer cells within the tumor have equal ability to propagate the tumor growth depending on continuing mutations and selective pressure. By contrast, the stem cells model suggests that cancer heterogeneity is due to the hierarchy that originates from a small population of cancer stem cells (CSCs) which are biologically distinct from the bulk tumor and possesses self-renewal, tumorigenic and multilineage potential. Although these two hypotheses have been discussed for a long time as mutually exclusive explanations of tumor heterogeneity, they are easily reconciled serving as a driving force of cancer evolution and diversity. Recent discovery of the cancer cell plasticity and heterogeneity makes the CSC population a moving target that could be hard to track and eradicate. Understanding the signaling mechanisms regulating CSCs during the course of cancer treatment can be indispensable for the optimization of current treatment strategies

Gender Identity, Disability, and Unmet Healthcare Needs among Disabled People Living in the Community in the United States

Disabled adults and transgender people in the United States face multiple compounding and marginalizing forces that result in unmet healthcare needs. Yet, gender identity among disabled people has not been explored, especially beyond binary categories of gender. Using cross-sectional survey data, we explored the rates of disability types and the odds of unmet healthcare needs among transgender people with disabilities compared to cisgender people with disabilities. The rates of disability type were similar between transgender and cisgender participants with two significant differences. Fewer transgender participants identified physical or mobility disability as their main disability compared to cisgender participants (12.31%/8 vs. 27.68/581, p < 0.01), and more transgender participants selected developmental disability as their main disability compared to cisgender participants (13.85%/9 vs. 3.67%/77, p < 0.001). After adjusting for sociodemographic characteristics, the odds of disabled transgender participants reporting an unmet need were higher for every unmet need except for preventative services

Fishery collapse, recovery, and the cryptic decline of wild salmon on a major California river

Fall-run Chinook salmon (Oncorhynchus tshawytscha) from the Sacramento–San Joaquin River system form the backbone of California’s salmon fishery and are heavily subsidized through hatchery production. Identifying temporal trends in the relative contribution of hatchery- versus wild-spawned salmon is vital for assessing the status and resiliency of wild salmon populations. Here, we reconstructed the proportion of hatchery fish on natural spawning grounds in the Feather River, a major tributary to the Sacramento River, using strontium isotope (87Sr/86Sr) ratios of otoliths collected during carcass surveys from 2002 to 2010. Our results show that prior to the 2007–2008 salmon stock collapse, 55%–67% of in-river spawners were of hatchery origin; however, hatchery contributions increased drastically (89%) in 2010 following the collapse. Data from a recent hatchery marking program corroborate our results, showing that hatchery fish continued to dominate (∼90%) in 2011–2012. Though the rebound in abundance of salmon in the Feather River suggests recovery of the stock postcollapse, our otolith chemistry data document a persistent decline of wild spawners, likely leading to the erosion of locally adapted Feather River salmon populations

One-step selection of Vaccinia virus-binding DNA aptamers by MonoLEX

<p>Abstract</p> <p>Background</p> <p>As a new class of therapeutic and diagnostic reagents, more than fifteen years ago RNA and DNA aptamers were identified as binding molecules to numerous small compounds, proteins and rarely even to complete pathogen particles. Most aptamers were isolated from complex libraries of synthetic nucleic acids by a process termed SELEX based on several selection and amplification steps. Here we report the application of a new one-step selection method (MonoLEX) to acquire high-affinity DNA aptamers binding Vaccinia virus used as a model organism for complex target structures.</p> <p>Results</p> <p>The selection against complete Vaccinia virus particles resulted in a 64-base DNA aptamer specifically binding to orthopoxviruses as validated by dot blot analysis, Surface Plasmon Resonance, Fluorescence Correlation Spectroscopy and real-time PCR, following an aptamer blotting assay. The same oligonucleotide showed the ability to inhibit <it>in vitro </it>infection of Vaccinia virus and other orthopoxviruses in a concentration-dependent manner.</p> <p>Conclusion</p> <p>The MonoLEX method is a straightforward procedure as demonstrated here for the identification of a high-affinity DNA aptamer binding Vaccinia virus. MonoLEX comprises a single affinity chromatography step, followed by subsequent physical segmentation of the affinity resin and a single final PCR amplification step of bound aptamers. Therefore, this procedure improves the selection of high affinity aptamers by reducing the competition between aptamers of different affinities during the PCR step, indicating an advantage for the single-round MonoLEX method.</p

Multimodal imaging techniques to evaluate the anticancer effect of cold atmospheric pressure plasma

Background: Skin cancer is the most frequent cancer worldwide and is divided into non-melanoma skin cancer, including basal cell carcinoma, as well as squamous cell carcinoma (SCC) and malignant melanoma (MM). Methods: This study evaluates the effects of cold atmospheric pressure plasma (CAP) on SCC and MM in vivo, employing a comprehensive approach using multi-modal imaging techniques. Longitudinal MR and PET/CT imaging were performed to determine the anatomic and metabolic tumour volume over three‐weeks in vivo. Additionally, the formation of reactive species after CAP treatment was assessed by non‐invasive chemiluminescence imaging of L‐012. Histological analysis and immunohistochemical staining for Ki‐67, ApopTag®, F4/80, CAE, and CD31, as well as protein expression of PCNA, caspase‐3 and cleaved‐caspase‐3, were performed to study proliferation, apoptosis, inflammation, and angiogenesis in CAP‐treated tumours. Results: As the main result, multimodal in vivo imaging revealed a substantial reduction in tumour growth and an increase in reactive species after CAP treatment, in comparison to untreated tu-mours. In contrast, neither the markers for apoptosis, nor the metabolic activity of both tumour entities was affected by CAP. Conclusions: These findings propose CAP as a potential adjuvant therapy option to established standard therapies of skin cancer

Analysis of reactive aldehydes in urine and plasma of type-2 diabetes mellitus patients through liquid chromatography-mass spectrometry: Reactive aldehydes as potential markers of diabetic nephropathy

IntroductionDiabetes is a major public health issue that is approaching epidemic proportions globally. Diabetes mortality is increasing in all ethnic groups, irrespective of socio-economic class. Obesity is often seen as the main contributor to an increasing prevalence of diabetes. Oxidative stress has been shown to trigger obesity by stimulating the deposition of white adipose tissue. In this study, we measured reactive aldehydes by liquid chromatography-mass spectrometry (LC-MS), in the urine and plasma of type-2 diabetic mellitus (T2DM) patients, as potential surrogates of oxidative stress. Our hypothesis was that reactive aldehydes play a significant role in the pathophysiology of diabetes, and these reactive species, may present potential drug targets for patient treatment.Materials and methodsStudy participants [N = 86; control n = 26; T2DM n = 32, and diabetic nephropathy (DN) n = 28] were recruited between 2019 and 2020. Urine and blood samples were collected from all participants, including a detailed clinical history, to include patient behaviours, medications, and co-morbidities. Reactive aldehyde concentrations in urine and plasma were measured using pre-column derivatisation and LC-MS, for control, T2DM and DN patients.ResultsReactive aldehydes were measured in the urine and plasma of control subjects and patients with T2DM and DN. In all cases, the reactive aldehydes under investigation; 4-HNE, 4-ONE, 4-HHE, pentanal, methylglyoxal, and glyoxal, were significantly elevated in the urine and serum of the patients with T2DM and DN, compared to controls (p &lt; 0.001) (Kruskal–Wallis). Urine and serum reactive aldehydes were significantly correlated (≥0.7) (p &lt; 0.001) (Spearman rho). The concentrations of the reactive aldehydes were significantly higher in plasma samples, when compared to urine, suggesting that plasma is the optimal matrix for screening T2DM and DN patients for oxidative stress.ConclusionReactive aldehydes are elevated in the urine and plasma of T2DM and DN patients. Reactive aldehydes have been implicated in the pathobiology of T2DM. Therefore, if reactive aldehydes are surrogates of oxidative stress, these reactive aldehyde species could be therapeutic targets for potential drug development

Comparison of BMI Derived from Parent-Reported Height and Weight with Measured Values: Results from the German KiGGS Study

The use of parent-reported height and weight is a cost-efficient instrument to assess the prevalence of children’s weight status in large-scale surveys. This study aimed to examine the accuracy of BMI derived from parent-reported height and weight and to identify potential predictors of the validity of BMI derived from parent-reported data. A subsample of children aged 2–17 years (n = 9,187) was taken from the 2003–2006 cross-sectional German KiGGS study. Parent-reported and measured height and weight were collected and BMI was calculated. Besides descriptive analysis, linear regression models with BMI difference and logistic regression models with weight status misclassification as dependent variables were calculated. Height differences varied by gender and were generally small. Weight and BMI were under-reported in all age groups, the under-reporting getting stronger with increasing age. Overall, the proportion for overweight and obesity based on parental and measured reports differed slightly. In the youngest age group, the proportion of overweight children was overestimated, while it was underestimated for older children and adolescents. Main predictors of the difference between parent reported and measured values were age, gender, weight status and parents’ perception of the child’s weight. In summary, the exclusive use of uncorrected parental reports for assessment of prevalence rates of weight status is not recommended