Miracle-dz/UniFAR-FTUs huggingface dataset:
A high-throughput assay for enzymatic polyester hydrolysis activity by fluorimetric detection.

A fluorimetric assay for the fast determination of the activity of polyester-hydrolyzing enzymes in a large number of samples has been developed. Terephthalic acid (TPA) is a main product of the enzymatic hydrolysis of polyethylene terephthalate (PET), a synthetic polyester. Terephthalate has been quantified following its conversion to the fluorescent 2-hydroxyterephthalate by an iron autoxidation-mediated generation of free hydroxyl radicals. The assay proved to be robust at different buffer concentrations, reaction times, pH values, and in the presence of proteins. A validation of the assay was performed by analyzing TPA formation from PET films and nanoparticles catalyzed by a polyester hydrolase from Thermobifida fusca KW3 in a 96-well microplate format. The results showed a close correlation (R(2) = 0.99) with those obtained by a considerably more tedious and time-consuming HPLC method, suggesting the aptness of the fluorimetric assay for a high-throughput screening for polyester hydrolases. The method described in this paper will facilitate the detection and development of biocatalysts for the modification and degradation of synthetic polymers. The fluorimetric assay can be used to quantify the amount of TPA obtained as the final degradation product of the enzymatic hydrolysis of PET. In a microplate format, this assay can be applied for the high-throughput screening of polyester hydrolases.
The impact of HAART on HPV-related cervical disease.

PURPOSE OF REVIEW highly active antiretroviral therapy (HAART) has had an unequivocally positive impact on morbidity and mortality in HIV-infected individuals. These benefits have clearly extended to some HIV-related malignancies, including Kaposi's sarcoma and non-Hodgkin's lymphoma. The impact of HAART on cervical cancer, however, remains uncertain. The objective of this review is to summarize the last ten years of registry-based and clinical research into the impact of HAART on human papillomavirus (HPV) related cervical disease. RELEVANT FINDINGS compared to their HIV-uninfected counterparts, HIV-infected women have an increased prevalence of HPV infection, increased risk of progression of HPV-related cervical disease, and an increased risk of invasive cervical cancer. While the partial immune reconstitution afforded by HAART might be expected to decrease susceptibility to HPV infection and cervical disease, the local effects of improved immunosurveillance on the cervix are uncertain and the increased longevity of patients on HAART may increase risk of exposure to HPV and provide the time required for progression of cervical disease. Registry-based evidence has been consistent in identifying the lack of decrease in cervical cancer incidence in the HAART era. Clinical research on the subject, however, has produced conflicting evidence with regards to both the effect of HAART on HPV infection and its impact on cervical disease progression/regression. SUMMARY the incidence of cervical cancer has not decreased in the HAART-era. Furthermore, clinical research has not shown a clear benefit of HAART in decreasing HPV-related cervical disease in HIV-infected women. A better understanding of this subject will have an impact on cervical disease surveillance practices.
Programas de transferencias condicionadas: balance de la experiencia reciente en América Latina y el Caribe

El presente documento tiene por objeto sintetizar la experiencia de America Latina y el Caribe con los programas de transferencias condicionadas, o "con corresponsabilidad" (PTC), a lo largo de mas de 15 anos. Durante este tiempo, estos programas han logrado consolidarse en los distintos paises de la region y se han difundido como un instrumento privilegiado de la politica contra la pobreza. Segun la Base de datos de programas de proteccion social no contributiva en America Latina y el Caribe de la CEPAL, los PTC operan en la actualidad en 18 paises de la region y benefician a mas de 25 millones de familias (alrededor de 113 millones de personas), es decir, el 19% de la poblacion de America Latina y el Caribe, a un costo que ronda el 0,4% del PIB regional. La estructura basica de los PTC consiste en la entrega de recursos monetarios y no monetarios a familias en situacion de pobreza o pobreza extrema que tienen hijos menores de edad, con la condicion de que estas cumplan con ciertos compromisos asociados al mejoramiento de sus capacidades humanas. A pesar de los debates aun inconclusos sobre la idoneidad de estos programas y sus resultados en distintos ambitos, se ha planteado que pueden representar un paso importante para poner a las familias pobres e indigentes con ninos en edad escolar en contacto con sistemas de proteccion social mas amplios e integrales. En este documento, que se espera sirva de base e insumo para la discusion y el avance hacia la construccion de sistemas de proteccion social fundados en las nociones de inclusion y universalidad de los derechos, se brinda informacion detallada sobre los distintos componentes de los PTC. Ademas, se revisan las principales caracteristicas de los programas en cuanto a la definicion y el registro de destinatarios, los mecanismos de focalizacion utilizados, los diversos tipos de prestaciones que ofrecen y las condicionalidades que los acompanan. Se analiza, asimismo, la evolucion historica de los indicadores de inversion y cobertura de los PTC, y la informacion disponible respecto de sus efectos en distintos ambitos. Por ultimo, se hace un balance de la experiencia y los principales desafios que estos programas presentan en materia de sostenibilidad, marco legal, rendicion de cuentas, participacion, institucionalidad e intersectorialidad.
Effects of experimental conditions on mycorrhizal relationships between Pinus sylvestris and Lactarius deliciosus and unprecedented fruit-body formation of the Saffron milk cap under controlled soilless conditions.

The mycorrhizal relationships between pines and two edible species of Lactarius sect. Dapetes were investigated by optimizing the experimental conditions of mycelial growth and of mycorrhizal colonization of pine seedlings. In vitro mycelial growth of Lactarius deliciosus and L. sanguifluus was improved on a buffered medium containing glucose, amino acids, and vitamins. Two methods of mycorrhization of pines with Lactarius deliciosus were tested. The mycorrhizal colonization was rapid and intense under non-aseptic conditions with a low nutrient supply and without exogenous glucose. A positive influence of mycorrhizal colonization on Pinus sylvestris growth was subsequently observed. Under axenic conditions and with a high nutrient supply, mycorrhization was stimulated at 10 g/L of exogenous glucose, irrespective of the phosphorus concentration. At high phosphorus level (1 mM) and 0.1, 1.0, or 10.0 g/L glucose, growth of Pinus sylvestris was reduced by inoculation. Stability and development of Pinus spp. /Lactarius deliciosus symbioses were assayed in a climatic chamber using containers filled with a synthetic substrate. Over a 2-year culture period, the root systems of the pine seedlings were heavily colonized by Lactarius deliciosus. One year following inoculation, Lactarius deliciosus fruit-body primordia appeared associated with Pinus sylvestris seedlings. Six months later, two mature basidiomata were obtained. This is the first report of soilless fruit-body formation of this edible mushroom.
Erlotinib after Failure of Gefitinib in Patients with Advanced Non-small Cell Lung Cancer Previously Responding to Gefitinib

Background: Gefitinib and erlotinib are two orally active epidermal growth factor receptor tyrosine kinase inhibitors used in the treatment of advanced non-small cell lung cancer (NSCLC). Gefitinib was the first one to become available in Europe in an expanded access program. When erlotinib granted approval, there were many patients who had progressed after receiving gefitinib and different lines of chemotherapy with no further treatment options; moreover, the results of clinical trials suggested possible significant differences in the mechanisms of sensitivity and in the efficacy between these two agents. Methods: Advanced NSCLC White patients, previously treated with almost two lines of chemotherapy and with gefitinib, obtaining a partial response or a stable disease with this agent, were treated with erlotinib after progression of disease to gefitinib. Only eight patients resulted eligible because of the unavailability of gefitinib after the closure of the expanded access program. Results: Four patients were women; histologic diagnosis was adenocarcinoma in four cases and bronchoalveolar carcinoma or unspecified NSCLC in two cases each. Median age was 70 years (range, 53–85). Seven out of eight patients had never smoked, one was a former smoker. We obtained two partial responses and three stable diseases with erlotinib with a median duration of response of 8 months. The median time to progression and overall survival were 5.9 and 14.6 months, respectively. Conclusion: Erlotinib seems to be a potential therapeutic option for treatment of selected advanced NSCLC patients after failure of gefitinib. Further studies are warranted to evaluate the molecular mechanisms behind this evidence and clarify how to select patients for treatment with epidermal growth factor receptor tyrosine kinase inhibitors.
Exploration for an Algorithm for Deintensification to Exclude the Retropharyngeal Site From Advanced Oropharyngeal Squamous Cell Carcinoma Treatment.

IMPORTANCE Understanding the drainage patterns to the retropharyngeal lymph nodes is an important consideration in oropharyngeal squamous cell carcinoma (OPSCC) because treatment of these nodes is related to increased morbidity. Prediction of these drainage patterns could not only help minimize treatment morbidity but also prevent failures in at-risk patients as deintensification trials are under way for this disease. OBJECTIVE To evaluate the prevalence of pathologic retropharyngeal adenopathy (RPA) in OPSCC relative to involvement of the oropharyngeal subsite, number of metastatic neck nodes, T classification, and N classification. DESIGN, SETTING, AND PARTICIPANTS We performed a retrospective review from January 1, 2003, through December 31, 2010, at an academic referral center of 205 previously untreated patients with pathologically confirmed, advanced-stage (III, IV) OPSCC. Data analysis was performed from January 1, 2013, through June 30, 2015. EXPOSURE Concurrent chemoradiotherapy. MAIN OUTCOMES AND MEASURES Radiologic evidence of pathologic RPA was tabulated and related to involvement of the oropharyngeal subsite, number of metastatic neck nodes, T classification, and N classification. RESULTS Of the 205 previously untreated patients (183 men; mean age, 56.1 years), pathologic RPA was identified in 37 (18.0%) of the 205 patients. Pathologic retropharyngeal lymph nodes were found in 12 (13.5%) of 89 patients with base of tongue cancers, 24 (22.0%) of 109 patients with tonsil cancers, and 1 (14.3%) of 7 patients with other oropharyngeal subsite cancers. Increasing prevalence of RPA was positively correlated with closer proximity to the posterior tonsillar pillar. A multivariable logistic regression model using the oropharyngeal subsite, involvement of the posterior tonsillar pillar, number of metastatic neck nodes, T classification, and N classification revealed that the number of metastatic neck nodes was statistically significant (odds ratio, 1.44; 95% CI, 1.20-1.71; P < .001). CONCLUSIONS AND RELEVANCE The prevalence of pathologic RPA in this cohort was 18.0%, and patients with multiple nodes had the highest risk of pathologic RPA, followed by involvement of the posterior tonsillar pillar. However, these data suggest that there is no clear algorithm that can be used for deintensification to exclude the retropharyngeal site from the treatment volume using extent of disease gathered from pretreatment imaging for patients with advanced-stage OPSCC.
A single bout of exhaustive exercise affects integrated baroreflex function after 16 days of head-down tilt.

We tested the hypothesis that one bout of maximal exercise performed 24 h before reambulation from 16 days of 6 degrees head-down tilt (HDT) could increase integrated baroreflex sensitivity. Isolated carotid-cardiac and integrated baroreflex function was assessed in seven subjects before and after two periods of HDT separated by 11 mo. On the last day of one HDT period, subjects performed a single bout of maximal cycle ergometry (exercise). Subjects did not exercise after the other HDT period (control). Carotid-cardiac baroreflex sensitivity was evaluated using a neck collar device. Integrated baroreflex function was assessed by recording heart rate (HR) and blood pressure (MAP) during a 15-s Valsalva maneuver (VM) at a controlled expiratory pressure of 30 mmHg. The ratio of change in HR to change in MAP (delta HR/ delta MAP) during phases II and IV of the VM was used as an index of cardiac baroreflex sensitivity. Baroreflex-mediated vasoconstriction was assessed by measuring the late phase II rise in MAP. Following HDT, carotid-cardiac baroreflex sensitivity was reduced (2.8 to 2.0 ms/mmHg; P = 0.05) as was delta HR/ delta MAP during phase II (-1.5 to -0.8 beats/mmHg; P = 0.002). After exercise, isolated carotid baroreflex activity and phase II delta HR/ delta MAP returned to pre-HDT levels but remained attenuated in the control condition. Phase IV delta HR/ delta MAP was not altered by HDT or exercise. The late phase II increase of MAP was 71% greater after exercise compared with control (7 vs. 2 mmHg; P = 0.041). (ABSTRACT TRUNCATED AT 250 WORDS)
Transactional Associations Among Teacher Support, Peer Social Preference, and Child Externalizing Behavior: A Four-Wave Longitudinal Study

The links between children's externalizing behaviors and two characteristics of children's social interactions within the classroom, namely, peer social preference and received support from the teacher, were studied in 570 children followed from their 2nd- to 3rd-grade years of elementary school. Data consisted of peer and teacher reports of externalizing behavior, sociometric “liked most” and “liked least” nominations, and teacher rated support. Results showed consistent paths from externalizing behavior to (low) peer social preference. Peer social preference, in turn, predicted decreases in externalizing behavior, even after taking teacher support into account. Teacher support was not consistently linked to the development of externalizing behavior across time. However, an indirect path from externalizing behavior, via (low) peer social preference, to lower levels of teacher support was found. These results were similar for girls and boys.
Coronin is involved in uptake of Mycobacterium bovis BCG in human macrophages but not in phagosome maintenance

By applying density gradient electrophoresis (DGE) to human macrophages infected with Mycobacterium bovis BCG, we were able to separate three different bacterial fractions representing arrested phagosomes, phagolysosomes and mycobacterial clumps. After further purification of the phagosomal population, we found that isolated phagosomes containing live BCG were arrested in maturation as they exhibited only low amounts of the lysosomal glycoprotein LAMP‐1 and processing of the lysosomal hydrolase cathepsin D was blocked. In addition, low amounts of MHC class I and class II molecules and the absence of HLA‐DM suggest sequestration of mycobacterial phagosomes from antigen‐processing pathways. We further investigated the involvement of the actin‐binding protein coronin in intracellular survival of mycobacteria and showed that human coronin, as well as F‐actin, were associated with early stages of mycobacterial phagocytosis but not with phagosome maintenance. Therefore, we conclude that the unique DGE migration pattern of arrested phagosomes is not as a result of retention of coronin, but that there are other proteins or lipids responsible for the block in maturation in human macrophages.
Electrothermally Actuated Silicon Carbide Tunable MEMS Resonators

This paper presents the fabrication and characterization of silicon carbide (SiC) flexural-mode structures able to operate as electrothermomechanical tunable resonators. Single- and double-clamped beams, as well as circular structures, have been fabricated with aluminium and platinum (Pt) top electrodes. Electrothermal excitation has been used for device actuation and resonant-frequency tuning and mixing. Circular structures (i.e., disks) have been shown to possess higher resonant frequencies and Q factors (up to ~23 000) compared to beams having similar dimensions. The tuning of the resonant frequency has been performed by varying the dc and ac components of the actuating voltage on SiC beams with u-shaped and slab Pt electrodes. When increasing the dc bias, the frequency-shift rates of about -11 000 and -1100 ppm/V are measured for the u-shaped and slab electrodes, respectively. When increasing the amplitude of the ac input, shift rates of about -1800 and -800 ppm/V are measured. In addition, measurements have shown that the frequency-shift rate increases with the ambient temperature. Electrothermal mixing has been performed by applying two actuating voltages with the sum or difference of their frequencies matching the fundamental resonance of the SiC structure. The tuning of the electrothermally mixed output signal has been demonstrated on a disk resonator.
Hepcidin as a predictive factor and therapeutic target in erythropoiesis-stimulating agent treatment for anemia of chronic disease in rats

Anemia of chronic disease is a multifactorial disorder, resulting mainly from inflammation-driven reticuloendothelial iron retention, impaired erythropoiesis, and reduced biological activity of erythropoietin. Erythropoiesis-stimulating agents have been used for the treatment of anemia of chronic disease, although with varying response rates and potential adverse effects. Serum concentrations of hepcidin, a key regulator of iron homeostasis, are increased in patients with anemia of chronic disease and linked to the pathogenesis of this disease, because hepcidin blocks cellular iron egress, thus limiting availability of iron for erythropoiesis. We tested whether serum hepcidin levels can predict and affect the therapeutic efficacy of erythropoiesis-stimulating agent treatment using a well-established rat model of anemia of chronic disease. We found that high pre-treatment hepcidin levels correlated with an impaired hematologic response to an erythropoiesis-stimulating agent in rats with anemia of chronic disease. Combined treatment with an erythropoiesis-stimulating agent and an inhibitor of hepcidin expression, LDN-193189, significantly reduced serum hepcidin levels, mobilized iron from tissue stores, increased serum iron levels and improved hemoglobin levels more effectively than did the erythropoiesis-stimulating agent or LDN-193189 monotherapy. In parallel, both the erythropoiesis-stimulating agent and erythropoiesis-stimulating agent/LDN-193189 combined reduced the expression of cytokines known to inhibit erythropoiesis. We conclude that serum hepcidin levels can predict the hematologic responsiveness to erythropoiesis-stimulating agent therapy in anemia of chronic disease. Pharmacological inhibition of hepcidin formation improves the erythropoiesis-stimulating agent’s therapeutic efficacy, which may favor a reduction of erythropoiesis-stimulating agent dosages, costs and side effects.
Essential residues in the polar loop region of subunit c of Escherichia coli F1F0 ATP synthase defined by random oligonucleotide-primed mutagenesis

The conserved, polar loop region of subunit c of the Escherichia coli F1F0 ATP synthase is postulated to function in the coupling of proton translocation through F0 to ATP synthesis in F1. We have used a random mutagenesis procedure to define the essential residues in the region. Oligonucleotide-directed mutagenesis was carried out with a random mixture of mutant oligonucleotides, the oligonucleotide mixture being generated by chemical synthesis by using phosphoramidite nucleotide stocks that were contaminated with the other three nucleotides. Thirty mutant genes coding single-amino-acid substitutions in the region between Glu-37 and Leu-45 of subunit c were tested for function by analyzing the capacity of plasmids carrying the mutant genes to complement a Leu-4----amber subunit c mutant. All substitutions at the conserved Arg-41 residue resulted in loss of oxidative phosphorylation, i.e., transformants could not grow on a succinate carbon source. The other conserved residues were more tolerant to substitution, although most substitutions did result in impaired growth on succinate. We conclude that Arg-41 is essential in the function of the polar loop and that the ensemble of other conserved residues collectively maintain an optimal environment required for that function.
Cloning of the crystalline cell wall protein gene of Bacillus licheniformis NM 105

A protein with a tetragonal pattern, defined as RS protein, was found on the wall surface of an alkaline phosphatase secretion-deficient mutant (NM 105) of Bacillus licheniformis 749/C. The protein was present on the wall surface of the exponential-growth-phase cells, but at the stationary growth phase it was overproduced and hypersecreted. This protein was precipitated to homogeneity from the culture fluid by 80% ammonium sulfate saturation and chilled acetone. The molecular mass of the protein was 98 kilodaltons, and it had a single subunit in a sodium dodecyl sulfate gel. Specific anti-RS antibody was generated in rabbits and used to immunolabel the RS protein on the cells at different growth phases. In early-exponential-growth-phase cells, the outside surface of the wall, the cytoplasm, and the inside surface of the cytoplasmic membrane were labeled. In stationary-growth-phase cells, the cytoplasm was poorly labeled, but the labeling on the outside surface of the wall was high. AB. licheniformis NM 105 gene library was made by using the lambda phage EMBL3. The RS protein expression from this gene library was detected by a modified autoradiographic procedure. One of the amplified RS protein-positive plaques (4213-1) containing recombinant DNA was chosen, and the restriction map of this DNA was prepared. The RS protein expressed in Escherichia coli NM 539 infected with 4213-1 recombinant phage had a lower molecular mass than the purified authentic RS protein. The 4.5-kilobase-pair (kbp) SalI-EcoRI fragment of the recombinant DNA was cloned in the shuttle plasmid pMK4 to construct pMK462, which was expressed in B. subtilis MI112 and produced the RS protein identical in molecular mass to the purified authentic RS protein. The RS protein expression was also demonstrated in cryosections of transformed E. coli and B. subtilis cells by immunoelectron microscopy. The 1.2-kbp SalI-HindIII and 1.8-kbp HindIII-HindIII recombinant DNA restriction enzyme fragments, respectively, from the right of the restriction map produced anti-RS antibody cross-reacting proteins. The expression of the 1.2-kbp SalI-HindIII DNA fragment cloned in pUC8 could be induced with isopropyl-beta-D-thiogalactopyranoside. The 1.8-kbp DNA restriction fragment hybridized with both the chromosomal DNA of strain NM 105 and the recombinant phage 4213-1 DNA. The RS gene expression was finally demonstrated in transformed E. coli 539 cells by in situ hybridization of frozen thin sections with the 1.8-kbp HindIII biotin-dATP probe and immunolabeling these with anti-biotin immunoglobulin G and protein A-gold.