Deregulation and epigenetic modification of BCL2-family genes cause resistance to venetoclax in hematologic malignancies.

The BCL2 inhibitor venetoclax has been approved to treat different hematological malignancies. Because there is no common genetic alteration causing resistance to venetoclax in chronic lymphocytic leukemia (CLL) and B-cell lymphoma, we asked if epigenetic events might be involved in venetoclax resistance. Therefore, we employed whole-exome sequencing, methylated DNA immunoprecipitation sequencing, and genome-wide clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 screening to investigate venetoclax resistance in aggressive lymphoma and high-risk CLL patients. We identified a regulatory CpG island within the PUMA promoter that is methylated upon venetoclax treatment, mediating PUMA downregulation on transcript and protein level. PUMA expression and sensitivity toward venetoclax can be restored by inhibition of methyltransferases. We can demonstrate that loss of PUMA results in metabolic reprogramming with higher oxidative phosphorylation and adenosine triphosphate production, resembling the metabolic phenotype that is seen upon venetoclax resistance. Although PUMA loss is specific for acquired venetoclax resistance but not for acquired MCL1 resistance and is not seen in CLL patients after chemotherapy-resistance, BAX is essential for sensitivity toward both venetoclax and MCL1 inhibition. As we found loss of BAX in Richter's syndrome patients after venetoclax failure, we defined BAX-mediated apoptosis to be critical for drug resistance but not for disease progression of CLL into aggressive diffuse large B-cell lymphoma in vivo. A compound screen revealed TRAIL-mediated apoptosis as a target to overcome BAX deficiency. Furthermore, antibody or CAR T cells eliminated venetoclax resistant lymphoma cells, paving a clinically applicable way to overcome venetoclax resistance.

Using numerical methods from nonlocal optics to simulate the dynamics of N-body systems in alternative theories of gravity.

The generalized Schrödinger-Newton system of equations with both local and nonlocal nonlinearities is widely used to describe light propagating in nonlinear media under the paraxial approximation. However, its use is not limited to optical systems and can be found to describe a plethora of different physical phenomena, for example, dark matter or alternative theories for gravity. Thus, the numerical solvers developed for studying light propagating under this model can be adapted to address these other phenomena. Indeed, in this work we report the development of a solver for the HiLight simulations platform based on GPGPU supercomputing and the required adaptations for this solver to be used to test the impact of new extensions of the Theory of General Relativity in the dynamics of the systems. In this work we shall analyze theories with nonminimal coupling between curvature and matter. This approach in the study of these new models offers a quick way to validate them since their analytical analysis is difficult. The simulation module, its performance, and some preliminary tests are presented in this paper.

Assessment of the Insecticidal Potential of the Eupatorium buniifolium Essential Oil Against Triatoma infestans (Hemiptera: Reduviidae). A Chiral Recognition Approach.

In this research, bioactivities toward the Chagas' disease vector Triatoma infestans (Klug) (Hemiptera: Reduviidae) by the essential oil (EO) of Eupatorium buniifolium H. et A. (Asteraceae) are reported. The tests were designed in order to determine ovicidal activity as well as the response to vapor exposure (fumigant) and to topical application (contact toxicity) and as repellent. In the last three bioassays, nymphs from the 3rd and 4th instar were used. The assayed materials were obtained from aerial parts of plants collected during the months of March and December, throughout 4 years, in two locations. The EO samples were subjected to a qualitative analysis by GC-MS and the relative area of each component was reported by GC-FID. The main monoterpene detected was α-pinene and by using a chiral column through GC-MS experiments and having both stereoisomers as standards, we were able to determine that the enantiomer present was S,S-(-)-α-pinene. Although usually in studies of EOs changes in chemical composition are often observed due to the time of collection and the environment where the plant develops, in our case the differences were, with some exception, only at the level of the minor components. The best results were obtained in the experiments to determine ovicidal activity, fumigant action, and repellency. No worthy response was found as insecticide in the trials designed for contact toxicity. The results of the studied bioactivities were independent of the location, month, and year of collection of the plant material. This behavior provides an interesting scope in relation to the potential use of this natural blend for the control of this insect at the nymph stage as repellent as well as for decreasing the population by ovicidal effect. Notably, in the course of the two-choice repellency test, it was possible to demonstrate recognition of one of the enantiomers of the α-pinene, giving rise to a non-common chirality/response effect. In this assay, the levorotatory isomer was the most active as repellent. Considering the abundance of the wild plant under study and the fact that its EO is easy to obtain, it is suggested that it could be an adequate natural resource to control this vector in a sustainable way as a complementary approach to conventional methods.

New potentiometric sensor based on molecularly imprinted nanoparticles for cocaine detection.

Here we present a potentiometric sensor for cocaine detection based on molecularly imprinted polymer nanoparticles (nanoMIPs) produced by the solid-phase imprinting method. The composition of polymers with high affinity for cocaine was optimised using molecular modelling. Four compositions were selected and polymers prepared using two protocols: chemical polymerisation in water and UV-initiated polymerisation in organic solvent. All synthesised nanoparticles had very good affinity to cocaine with dissociation constants between 0.6nM and 5.3nM. Imprinted polymers produced in organic solvent using acrylamide as a functional monomer demonstrated the highest yield and affinity, and so were selected for further sensor development. For this, nanoparticles were incorporated within a PVC matrix which was then used to prepare an ion-selective membrane integrated with a potentiometric transducer. It was demonstrated that the sensor was able to quantify cocaine in blood serum samples in the range of concentrations between 1nM and 1mM.

Development of molecularly imprinted polymers specific for blood antigens for application in antibody-free blood typing.

A novel approach in antibody-free blood typing based on molecularly imprinted polymeric nanoparticles is described.

Formation of target-specific binding sites in enzymes: solid-phase molecular imprinting of HRP.

Here we introduce a new concept for synthesising molecularly imprinted nanoparticles by using proteins as macro-functional monomers. For a proof-of-concept, a model enzyme (HRP) was cross-linked using glutaraldehyde in the presence of glass beads (solid-phase) bearing immobilized templates such as vancomycin and ampicillin. The cross-linking process links together proteins and protein chains, which in the presence of templates leads to the formation of permanent target-specific recognition sites without adverse effects on the enzymatic activity. Unlike complex protein engineering approaches commonly employed to generate affinity proteins, the method proposed can be used to produce protein-based ligands in a short time period using native protein molecules. These affinity materials are potentially useful tools especially for assays since they combine the catalytic properties of enzymes (for signaling) and molecular recognition properties of antibodies. We demonstrate this concept in an ELISA-format assay where HRP imprinted with vancomycin and ampicillin replaced traditional enzyme-antibody conjugates for selective detection of templates at micromolar concentrations. This approach can potentially provide a fast alternative to raising antibodies for targets that do not require high assay sensitivities; it can also find uses as a biochemical research tool, as a possible replacement for immunoperoxidase-conjugates.

Hydrogen sensing via anomalous optical absorption of palladium-based metamaterials.

A palladium (Pd)-based optical metamaterial has been designed, fabricated and characterized for its application in hydrogen sensing. The metamaterial can replace Pd thin films in optical transmission schemes for sensing with performances far superior to those of conventional sensors. This artificial material consists of a palladium-alumina metamaterial fabricated using inexpensive and industrial-friendly bottom-up techniques. During the exposure to hydrogen, the system exhibits anomalous optical absorption when compared to the well-known response of Pd thin films, this phenomenon being the key factor for the sensor sensitivity. The exposure to hydrogen produces a large variation in the light transmission through the metamembrane (more than 30% with 4% in volume hydrogen-nitrogen gas mixture at room temperature and atmospheric pressure), thus avoiding the need for sophisticated optical detection systems. An optical homogenization model is proposed to explain the metamaterial response. These results contribute to the development of reliable and low-cost hydrogen sensors with potential applications in the hydrogen economy and industrial processes to name a few, and also open the door to optically study the hydrogen diffusion processes in Pd nanostructures.

Does size matter? Study of performance of pseudo-ELISAs based on molecularly imprinted polymer nanoparticles prepared for analytes of different sizes.

The aim of this work is to evaluate whether the size of the analyte used as template for the synthesis of molecularly imprinted polymer nanoparticles (nanoMIPs) can affect their performance in pseudo-enzyme linked immunosorbent assays (pseudo-ELISAs). Successful demonstration of a nanoMIPs-based pseudo-ELISA for vancomycin (1449.3 g mol(-1)) was demonstrated earlier. In the present investigation, the following analytes were selected: horseradish peroxidase (HRP, 44 kDa), cytochrome C (Cyt C, 12 kDa) biotin (244.31 g mol(-1)) and melamine (126.12 g mol(-1)). NanoMIPs with a similar composition for all analytes were synthesised by persulfate-initiated polymerisation in water. In addition, core-shell nanoMIPs coated with polyethylene glycol (PEG) and imprinted for melamine were produced in organics and tested. The polymerisation of the nanoparticles was done using a solid-phase approach with the correspondent template immobilised on glass beads. The performance of the nanoMIPs used as replacement for antibodies in direct pseudo-ELISA (for the enzymes) and competitive pseudo-ELISA for the smaller analytes was investigated. For the competitive mode we rely on competition for the binding to the nanoparticles between free analyte and corresponding analyte-HRP conjugate. The results revealed that the best performances were obtained for nanoMIPs synthesised in aqueous media for the larger analytes. In addition, this approach was successful for biotin but completely failed for the smallest template melamine. This problem was solved using nanoMIP prepared by UV polymerisation in an organic media with a PEG shell. This study demonstrates that the preparation of nanoMIP by solid-phase approach can produce material with high affinity and potential to replace antibodies in ELISA tests for both large and small analytes. This makes this technology versatile and applicable to practically any target analyte and diagnostic field.

Optomechanical entanglement between a movable mirror and a cavity field.

We show how stationary entanglement between an optical cavity field mode and a macroscopic vibrating mirror can be generated by means of radiation pressure. We also show how the generated optomechanical entanglement can be quantified, and we suggest an experimental readout scheme to fully characterize the entangled state. Surprisingly, such optomechanical entanglement is shown to persist for environment temperatures above 20 K using state-of-the-art experimental parameters.

Fluorescence study of dehydroabietic acid-based bipolar arylamine-quinoxalines.

The absorption and fluorescence spectra, lifetimes and quantum yields of a series of triarylaminequinoxaline bipolar compounds, with and without the bulky dehydroabietic acid group, have been studied in toluene solution. This bulky group is introduced to improve solubility and thermal properties of these systems. It is shown that this does not affect their spectral or photophysical behavior. The compounds show relatively strong fluorescence, with the emission maximum strongly dependent upon the substituents present. Oxidation potentials have also been determined in acetonitrile solution, and again indicate that introduction of the resin acid moiety has no effect on these properties.

Helicobacter pylori virulence genotypes in Portuguese children and adults with gastroduodenal pathology.

The aim of this study was to evaluate the prevalence of virulence genotypes, namely cagA, vacA and babA2, of Helicobacter pylori strains isolated from Portuguese adults and children presenting gastroduodenal pathology. One hundred thirty-six strains were studied, 82 isolated from adult patients (50 with nonulcerative gastritis and 32 with active peptic ulcer) and 58 isolated from children (54 with nonulcerative gastritis and 4 with duodenal ulcer). Genotyping of cagA, vacA and babA2 was assessed by polymerase chain reaction. Overall, Helicobacter pylori strains carrying more virulent genotypes were much more prevalent in adults than in children, particularly the type I ( vacAs1- and cagA-positive) and the triple-positive ( vacAs1-, cagA- and babA2-positive) strains ( P<0.001). A subpopulation of adults and children with nonulcerative gastritis was also studied, and differences in the prevalence of virulent genotypes were observed, either for individual genotypes ( P=0.017 for cagA, P=0.010 for vacAs1) or in combinations, i.e. the type I genotype ( P=0.005) and the triple-positive strains ( P=0.031). There was no difference between the two populations in the distribution of babA2 and m1/m2 genotypes. Considering the cohort effect in the epidemiology of Helicobacter pylori infection, these results suggest that different strains might circulate during different periods of time, or that, after infection in childhood, individual strains will undergo changes during the course of infection.

[Endoprosthesis in the treatment of severe abdominal angina]. / Endoprótese no tratamento de angina abdominal grave.

The endovascular treatment (percutaneous transluminal angioplasty and stent placement) is an option to surgical treatment in patients with abdominal angor, with good results by a less invasive way. A case of a patient with superior mesenteric artery occlusion and severe stenosis of celiac axis is presented that was treated by endovascular treatment and stenting, with resolution of the abdominal angor.

Gastric cancer (carcinoma) and Helicobacter pylori: situation in Portugal.

This article of review describes the gastric cancer and Helicobacter pylori situation in Portugal. Regarding gastric carcinoma, it refers to the incidence data, geographical distribution as well as nutritional factors, which are potentially cancerous. Also mentioned are the prevalence data of Helicobacter pylori in Portugal.

Features and trends in Helicobacter pylori antibiotic resistance in Lisbon area, Portugal (1990-1999).

The features of Helicobacter pylori antibiotic resistance in Lisbon from 1990 to 1999 were studied. Overall resistance rates to amoxycillin, tetracycline, metronidazole, clarithromycin and ciprofloxacin were 0, 0, 30.6, 19.0 and 9.6%, respectively. The incidence of resistance to clarithromycin was much higher in isolates from children (44.8%) than adults (14.6%). For metronidazole, the contrary was observed (children: 19.0%, adults: 32.3%). Ciprofloxacin-resistant isolates were all from adult patients. Concerning the adult population, the resistance rate to metronidazole showed a slight increase during the decade, while for clarithromycin and ciprofloxacin a significant increase was observed (4.6 to 22.0% and 0 to 20.9%, respectively).

Field quantization in a plasma: photon mass and charge

It is shown here that a straightforward procedure can be used to quantize the linearized equations for an electromagnetic field in a plasma. This leads to a definition of an effective mass for the transverse photons, and a different one for the longitudinal photons, or plasmons. Both masses are simply proportional to the electron plasma density. A nonlinear perturbative analysis can also be used to extend the quantization procedure, in order to include the ponderomotive force effects. This leads to the definition of a photon charge operator. The mean value of this operator, for a quantum state with a photon occupation number equal to 1, is the equivalent charge of the photon in a plasma.

[Antineutrophil cytoplasmic antibodies in patients with sickle cell anemia]. / Anticuerpos anti citoplasma de neutrófilos en pacientes con anemia drepanocítica.

PURPOSE: To determine whether anti neutrophil cytoplasmic antibodies (ANCA): anti myeloperoxidase (anti MPO) and anti proteinase 3 (anti PR3), are present in patients with sickle cell anaemia (SCA) during painful crisis. PATIENTS AND METHODS: 55 patients with SCA were included in this study, 35 of them had painful crisis and the rest were in steady state. We determined levels of anti MPO and anti PR3 by two enzyme immunoassays commercially available (Progen Biotechnik GMBH). RESULTS: A significant increase of anti MPO in the group of patients with painful crisis was found in comparison with steady state patients and controls. CONCLUSIONS: Our results suggest the possibility that anti MPO interacting with MPO on primed neutrophils lead to neutrophil activation, enhanced neutrophil adhesion and amplification of endothelial cell damage.

[Pure autonomic failure]. / Falência autonómica pura.

We present the case of a 64-year-old woman who, in the past 5 years, complained of constipation/diarrhea, hyposudoresis, xerostomia and xerophthalmia, dysuria and orthostatic hypotension. Cardiovascular reflexes analysis revealed sympathetic and parasympathetic failure. Norepinephrine was markedly reduced, both lying and after tilt. Norepinephrine infusion determined a significant rise in blood pressure, allowing the diagnosis of denervation hypersensitivity. The diagnosis of pure autonomic failure was made. Therapy with 9 alpha fludrocortisone and metoclopramide was initiated with marked and sustained symptomatic effect.

Omeprazole in the treatment of peptic ulcers resistant to H2-receptor antagonists.

Thirty patients with peptic ulcers resistant to at least 8 weeks of continuous therapy with full-dose H2-receptor antagonists alone or followed by other anti-ulcer drugs, were treated with the gastric proton pump inhibitor omeprazole (40 mg), administered orally once daily for up to 8 weeks. The study design was non-comparative and open; healing was verified by endoscopy. After only 2 weeks of treatment, 21 out of 23 (91%) duodenal ulcer patients were healed, as well as 2 out of 2 patients with both duodenal and gastric ulcer and 1 out of 3 patients with prepyloric ulcer. After 4 weeks, all duodenal ulcers, 1 out of 2 gastric ulcers and 2 out of 3 pre-pyloric ulcers were healed. A further month of therapy healed the gastric ulcer to give an overall healing rate of 97% and leaving only one patient (pre-pyloric ulcer) unhealed at the end of the study. Of 19 patients suffering ulcer symptoms at entry, only two patients reported any symptoms at 2 weeks and one of these (who remained unhealed) continued to have symptoms throughout the study. One patient reported mild asthenia; otherwise, no clinical or biochemical side-effects were recorded. It is concluded that omeprazole is highly effective in healing refractory peptic ulcers.