Improving the Management of Endometrial Cancer Patients through the Use of Liquid Biopsy Analyses: A Case Report

Endometrial cancer (EC) is the 4th most common neoplasm of the female genital tract, with 15–20% of patients being of high risk of recurrence which leads to a significant decrease in patient survival. Current therapeutic options for patients with EC are poor, being the combined therapy of carboplatin and paclitaxel the standard of care, with limited efficacy. Therefore, new therapeutic options and better monitoring tools are needed to improve the management of the disease. In the current case report, we showcase the value of liquid biopsy analyses in a microsatellite instability EC patient with initially good prognosis that however underwent rapid progression disease within 6 months post-surgery; through the study of plasma cfDNA/ctDNA dynamics to assess the tumour evolution during treatment, as well as the study of the uterine aspirate as a valuable sample that captures the intra-tumour heterogeneity that allows a comprehensive genomic profiling of the disease to identify potential therapeutic options. Furthermore, preclinical models were generated at the time of tumour progression to assess the efficacy of the identified targeted therapies.     

Facile Microwave Assisted Synthesis of Silver Nanostars for Ultrasensitive Detection of Biological Analytes by SERS

We report a very simple, rapid and reproducible method for the fabrication of anisotropic silver nanostars (AgNS) that can be successfully used as highly efficient SERS substrates for different bioanalytes, even in the case of a near-infra-red (NIR) excitation laser. The nanostars have been synthesized using the chemical reduction of Ag⁺ ions by trisodium citrate. This is the first research reporting the synthesis of AgNS using only trisodium citrate as a reducing and stabilizing agent. The key elements of this original synthesis procedure are rapid hydrothermal synthesis of silver nanostars followed by a cooling down procedure by immersion in a water bath. The synthesis was performed in a sealed bottom flask homogenously heated and brought to a boil in a microwave oven. After 60 s, the colloidal solution was cooled down to room temperature by immersion in a water bath at 35 °C. The as-synthesized AgNS were washed by centrifugation and used for SERS analysis of test molecules (methylene blue) as well as biological analytes: pharmaceutical compounds with various Raman cross sections (doxorubicin, atenolol & metoprolol), cell lysates and amino acids (methionine & cysteine). UV-Vis absorption spectroscopy, (Scanning) Transmission Electron Microscopy ((S)TEM) and Atomic Force Microscopy (AFM) have been employed for investigating nanostars’ physical properties.     

Lightweight Bytecode Verification

In this paper, we provide a theoretical foundation for and improvements to the existing bytecode verification technology, a critical component of the Java security model, for mobile code used with the Java “micro edition” (J2ME), which is intended for embedded computing devices. In Java, remotely loaded “bytecode” class files are required to be bytecode verified before execution, that is, to undergo a static type analysis that protects the platform's Java run-time system from so-called type confusion attacks such as pointer manipulation. The data flow analysis that performs the verification, however, is beyond the capacity of most embedded devices because of the memory requirements that the typical algorithm will need. We propose to take a proof-carrying code approach to data flow analysis in defining an alternative technique called “lightweight analysis” that uses the notion of a “certificate” to reanalyze a previously analyzed data flow problem, even on poorly resourced platforms. We formally prove that the technique provides the same guarantees as standard bytecode safety verification analysis, in particular that it is “tamper proof” in the sense that the guarantees provided by the analysis cannot be broken by crafting a “false” certificate or by altering the analyzed code. We show how the Java bytecode verifier fits into this framework for an important subset of the Java Virtual Machine; we also show how the resulting “lightweight bytecode verification” technique generalizes and simulates the J2ME verifier (to be expected as Sun's J2ME “K-Virtual machine” verifier was directly based on an early version of this work), as well as Leroy's “on-card bytecode verifier,” which is specifically targeted for Java Cards.     

Genetics and Epigenetics of Spontaneous Intracerebral Hemorrhage

Intracerebral hemorrhage (ICH) is a complex and heterogeneous disease, and there is no effective treatment. Spontaneous ICH represents the final manifestation of different types of cerebral small vessel disease, usually categorized as: lobar (mostly related to cerebral amyloid angiopathy) and nonlobar (hypertension-related vasculopathy) ICH. Accurate phenotyping aims to reflect these biological differences in the underlying mechanisms and has been demonstrated to be crucial to the success of genetic studies in this field. This review summarizes how current knowledge on genetics and epigenetics of this devastating stroke subtype are contributing to improve the understanding of ICH pathophysiology and their potential role in developing therapeutic strategies.     

Chronic Hyperkaliemia in Chronic Kidney Disease: An Old Concern with New Answers

Increasing potassium intake ameliorates blood pressure (BP) and cardiovascular (CV) prognoses in the general population; therefore the World Health Organization recommends a high-potassium diet (90–120 mEq/day). Hyperkalaemia is a rare condition in healthy individuals due to the ability of the kidneys to effectively excrete dietary potassium load in urine, while an increase in serum K⁺ is prevalent in patients with chronic kidney disease (CKD). Hyperkalaemia prevalence increases in more advanced CKD stages, and is associated with a poor prognosis. This scenario generates controversy on the correct nutritional approach to hyperkalaemia in CKD patients, considering the unproven link between potassium intake and serum K⁺ levels. Another concern is that drug-induced hyperkalaemia leads to the down-titration or withdrawal of renin-angiotensin system inhibitors (RASI) and mineralocorticoids receptors antagonists (MRA) in patients with CKD, depriving these patients of central therapeutic interventions aimed at delaying CKD progression and decreasing CV mortality. The new K⁺-binder drugs (Patiromer and Sodium-Zirconium Cyclosilicate) have proven to be adequate and safe therapeutic options to control serum K⁺ in CKD patients, enabling RASI and MRA therapy, and possibly, a more liberal intake of fruit and vegetables.     

Antitumor Effects of Ral-GTPases Downregulation in Glioblastoma

Glioblastoma (GBM) is the most common tumor in the central nervous system in adults. This neoplasia shows a high capacity of growth and spreading to the surrounding brain tissue, hindering its complete surgical resection. Therefore, the finding of new antitumor therapies for GBM treatment is a priority. We have previously described that cyclin D1-CDK4 promotes GBM dissemination through the activation of the small GTPases RalA and RalB. In this paper, we show that RalB GTPase is upregulated in primary GBM cells. We found that the downregulation of Ral GTPases, mainly RalB, prevents the proliferation of primary GBM cells and triggers a senescence-like response. Moreover, downregulation of RalA and RalB reduces the viability of GBM cells growing as tumorspheres, suggesting a possible role of these GTPases in the survival of GBM stem cells. By using mouse subcutaneous xenografts, we have corroborated the role of RalB in GBM growth in vivo. Finally, we have observed that the knockdown of RalB also inhibits cell growth in temozolomide-resistant GBM cells. Overall, our work shows that GBM cells are especially sensitive to Ral-GTPase availability. Therefore, we propose that the inactivation of Ral-GTPases may be a reliable therapeutic approach to prevent GBM progression and recurrence.     

Nutritional Calcium Supply Dependent Calcium Balance,Bone Calcification and Calcium Isotope Ratios in Rats

Serum calcium isotopes (δ^44/42Ca) have been suggested as a non-invasive and sensitive Ca balance marker. Quantitative δ^44/42Ca changes associated with Ca flux across body compartment barriers relative to the dietary Ca and the correlation of δ^44/42Ca_Serum with bone histology are unknown. We analyzed Ca and δ^44/42Ca by mass-spectrometry in rats after two weeks of standard-Ca-diet (0.5%) and after four subsequent weeks of standard- and of low-Ca-diet (0.25%). In animals on a low-Ca-diet net Ca gain was 61 ± 3% and femur Ca content 68 ± 41% of standard-Ca-diet, bone mineralized area per section area was 68 ± 15% compared to standard-Ca-diet. δ^44/42Ca was similar in the diets, and decreased in feces and urine and increased in serum in animals on low-Ca-diet. δ^44/42Ca_Bone was higher in animals on low-Ca-diet, lower in the diaphysis than the metaphysis and epiphysis, and unaffected by gender. Independent of diet, δ^44/42Ca_Bone was similar in the femora and ribs. At the time of sacrifice, δ^44/42Ca_Serum inversely correlated with intestinal Ca uptake and histological bone mineralization markers, but not with Ca content and bone mineral density by µCT. In conclusion, δ^44/42Ca_Bone was bone site specific, but mechanical stress and gender independent. Low-Ca-diet induced marked changes in feces, serum and urine δ^44/42Ca in growing rats. δ^44/42Ca_Serum inversely correlated with markers of bone mineralization.     

Variation in fatty acid composition of ewes' colostrum and mature milk fat

Temporal content variations of approximately 70 C4–C24 fatty acids (FA) in colostrum and milk of ewes consuming winter diet were determined by gas chromatography. The content of monounsaturated, polyunsaturated and straight-chain saturated FA 14:0–16:0 in colostrum was higher whereas the content of 4:0–12:0, 18:0 and branched-chain saturated FA 15:0–17:0 was lower than that in mature milk. The effect of ewe breed on the FA profiles was not significant. The composition of FA changed most significantly 1–2 days after lambing in agreement with the time schedule of colostrum formation. Nevertheless, the content of FA further successively changed up to approximately the 6th day of lactation, whereas further changes up to the 60th day of lactation were smaller. This is consistent with the recommendation that ewes' milk is suitable for human consumption after six-to-eight days of lactation. The higher contents of palmitic and myristic acids in colostrum compared with mature milk suggest that FA composition in colostrum matches the changing needs of the growing lambs.     

Measurement of Helium Leakage Rates through Closures of Dangerous Goods Packagings for the Assessment of Potentially Explosive Mixtures in Freight Containers

The objective was to find out whether an explosive atmosphere can be created in a freight container by gaseous leakage flow of vapour‐air‐mixture through leaks in the closures of dangerous goods packagings filled with hazardous liquids. Because of high temperatures during intercontinental carriage, there is a gauge pressure in the free vapour phase inside the packagings which can cause a gaseous leakage flow. Two different methods were applied: Helium limit leakage rates for 23 quantitatively important hazardous liquids concerning their lower explosion limit (LEL) were calculated for a worst case transport scenario (Method 1). Helium leakage rates of five closure types of dangerous goods packagings with volumes of approximately 6 l were measured using the pressure technique by accumulation (Method 2). All types of closures of steel packagings were uncritical. The maximum measured leakage was 33% of the limit leakage rate. The leakage rates of screw closures of plastic jerricans can exceed the LEL if there are production‐related patterns such as non‐concentricity of the closures and flashes on the neck. Especially for plastic packagings it is important to minimize gaseous leakage flow, because an explosive atmosphere can also be reached by permeation of the individual filling substance or by a combination of both effects. For the assessment of potentially explosive mixtures in freight containers, both mass transfer mechanisms have to be taken into account. Copyright © 2015 John Wiley & Sons, Ltd.