Tropane alkaloid production in transformed root cultures of brugmansia Candida

Transformed root cultures of BRUGMANSIA CANDIDA were established by infection with AGROBACTERIUM RHIZOGENES LBA 9402. Several clones with different growth index and tropane alkaloid pattern and content were obtained and two were examined in depth. The alkaloid content and pattern changed during the time course. At 21 days of culture clone 1 revealed an alkaloid spectrum dominated by 3alpha-acetoxytropane (about 50% of the total alkaloid) and hyoscyamine (about 25%), with a ratio of hyoscyamine to scopolamine of 11.2. In clone 40 this ratio was 1.5 and the content of 3alpha-acetoxytropane was low (2%). The maximum concentrations of hyoscyamine were obtained at 3 weeks of culture, and were 700 and 500 microg/g FW in clone 1 and 40, respectively.     

In vitro effects of resveratrol on oxidative stress in diabetic platelets

To evaluate the in vitro effects of resveratrol (RSV) incubation on platelets from compensated and decompensated diabetic patients in order to use it as an adjuvant therapy. The study was performed on 77 diabetic patients and divided into two phases: 29 compensated and 48 decompensated diabetic platelets were analyzed at recruitment (T ₀) and after in vitro RSV incubation (20 μg/ml) for 3 h at 37 °C (T ₁). Lipoperoxide and nitric oxide (NO) levels, superoxide dismutase (SOD) and Na⁺/K⁺ ATPase activities, total antioxidant capacity (TAC), and membrane fluidity tested by anisotropy of fluorescent probes TMA-DPH and DPH were determined. In vitro RSV incubation counteracts oxidative damage associated with diabetes and its complications; it is able to improve platelet function through augmented membrane fluidity and Na⁺/K⁺ ATPase activity; it enhances antioxidant systems’ functionality by increasing NO levels, SOD activity, and TAC and by decreasing lipoperoxide levels in both compensated and decompensated patients. Such platelet functionality enhancement suggests a new method of secondary prevention of complications associated with platelet dysfunction. Being free from one of the major risks associated with many antidiabetic agents, it can be assumed that RSV utilization in the diabetic diet may have a preventive and protective role in the progression of diabetic oxidative damage.     

Stimulated chemiluminescence of polymorphonuclear leukocytes in rheumatoid arthritis: in vivo enhancement by auranofin

The aim of the study was to investigate the interaction of opsonized zymosan with polymorphonuclear leukocytes (PMNLs) from 26 rheumatoid arthritis patients (six males and 20 females; mean age, 51.88 +/- 11.15 years; range, 33-72 years) and 11 control subjects (two males and nine females; mean age, 56.54 +/- 12.1 years; range, 33-75 years) using a luminol-amplified chemiluminescence (CL). The data obtained suggest that the opsonized zymosan stimulated CL is decreased in untreated rheumatoid patients compared with control subjects. The decreased stimulated CL may be the result of a physico-chemical alteration of the membrane (including decreased membrane fluidity and translocation of receptor sites) induced by free radicals of oxygen and/or by multiple exposures to chemotaxins. Auranofin enhances stimulated CL although the mechanism is not clear. Auranofin could interact with membrane lipids, induce changes in membrane surface charges and increase membrane fluidity. Our results are difficult to compare with those of previous studies, since different assays, and chemoattractants were applied. In addition, the disease activity and medication were also different.     

Cross-link immunoprecipitation data to detect polymorphisms lying in splicing regulatory motifs: a method to refine single nucleotide polymorphism selection in association studies

In a previous study we showed that prediction tools are useful to select single-nucleotide polymorphisms (SNPs) which potentially affect phenotype and therefore guide genotyping in association studies, thus saving time and money. Here we use the recently available RNA cross-link immunoprecipitation data to analyze several genes involved in psychiatric disorders and show which disease-associated SNPs can affect the splicing process by altering splicing factor binding sites. We point out the importance of using cross-link immunoprecipitation data in psychiatry to refine the SNP selection methods, to explain the association found and to plan molecular investigations.     

Artificial Neural Networks as a Way to Predict Future Kidney Cancer Incidence in the United States

IntroductionThe incidence of kidney cancer is increasing; it could be counteracted with new ways to predict and detect it. We aimed to implement an artificial neural network in order to predict new cases of renal-cell carcinoma (RCC) in the population using population rate, obesity, smoking incidence, uncontrolled hypertension, and life expectancy data in the United States.Patients and MethodsStatistics were collected on US population numbers, life expectancy, obesity, smoking, and hypertension. We used MATLAB R2018 (MathWorks) software to implement an artificial neural network. Data were repeatedly and randomly divided into training (70%) and validation (30%) subsets.ResultsThe number of new RCC cases will grow from 44,400 (2020) to 55,400 (2050), an increase of +24.7%. Our data show that preventing hypertension would have the greatest impact on reduction of the incidence, estimated at ?775 and ?575 cases per year in 2020 and in 2030, respectively. The prevention of obesity and smoking would have a more limited impact, estimated at ?64 and ?180 cases per year in 2020 and in 2030, respectively, for obesity, and ?173 and ?21 cases per year in 2020 and in 2030, respectively, for smoking.ConclusionsOur predictions underline the need for accurate studies on RCC-related risk factors to reduce the incidence.     

Three-dimensional nuclear architecture distinguishes thyroid cancer histotypes

Molecular markers can serve as diagnostic tools to support pathological analysis in thyroid neoplasms. However, because the same markers can be observed in some benign thyroid lesions, additional approaches are necessary to differentiate thyroid tumor subtypes, prevent overtreatment and tailor specific clinical management. This applies particularly to the recently described variant of thyroid cancer referred to as noninvasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP). This variant has an estimated prevalence of 4.4% to 9.1% of all papillary thyroid carcinomas worldwide. We studied 60 thyroid lesions: 20 classical papillary thyroid carcinoma (CPTC), 20 follicular variant of PTC (FVPTC) and 20 NIFTP. We examined morphological and molecular features to identify parameters that can differentiate NIFTP from the other PTC subtypes. When blindly investigating the nuclear architecture of thyroid neoplasms, we observed that NIFTP has significantly longer telomeres than CPTC and FVPTC. Super-resolved 3D-structured illumination microscopy demonstrated that NIFTP is heterogeneous and that its nuclei contain more densely packed DNA and smaller interchromatin spaces than CPTC and FVPTC, a pattern that resembles normal thyroid tissue. These data are consistent with the observed indolent biological behavior and favorable prognosis associated with NIFTP, which lacks BRAF^V600E mutations. Of note, next-generation thyroid oncopanel sequencing was unable to distinguish the thyroid cancer histotypes in our study cohort. In summary, our data suggest that 3D nuclear architecture can be a powerful analytical tool to diagnose and guide clinical management of NIFTP.