Search

7565 Background: Recurring mutations in epigenetic functions in PTCL, coupled with marked activity of epigenetic drugs, raises a question regarding whether these mutations might portend greater vulnerability to one drug over another. For example, do mutations in genes governing DNA methylation suggest these patients might benefit from a hypomethylating (HMA) agent? Preclinical data from our group suggests marked synergism between histone deacetylase inhibitors (HDACi) and HMA, as well as HDACi and pralatrexate (PDX), irrespective of mutations in epigenetic genes. Phase 1 studies (romidepsin [R] plus PDX or R plus 5-azacytidine [Aza]) are completed, and the Phase 2 studies are near completion. This clinical trial scenario affords a unique opportunity to decipher the impact of a HMA on response as a function of TET2, IDH2, DNMT3 and other mutations in PTCL. Methods: Patients with R/R lymphoma were eligible for the phase 1, whereas the phase 2 only enrolled patients with PTCL, either R/R or treatment-naïve individuals. Exploratory endpoints included next generation sequencing (NGS) and methylation arrays. Results: In toto, 89 patients have been enrolled on both trials across all histology’s, 58 have PTCL. NGS and efficacy data is available for the majority of patients, with some from the PDX + R study in process. The ORR among the PTCL patients for PDX+R and Aza+R has been 71% and 73% respectively. Eight of 9 angioimmunoblastic TCL patients responded. Among those with TET2 mutations, 7 of 8 responded to the Aza based treatment, while only 3 of 6 (50%) who were TET2 negative responded. Similarly, of 3 patients with DNMT3 treated with Aza based therapy, all 3 responded. Remarkably one patient with a TET2 mutation experienced progression to PDX+R, and CR to Aza+R. Among the PTCL patients treated with Aza, the global demethylation score (GDMS) demonstrated marked demethylation among all patients, with no correlation between the score and likelihood of response. Conclusions: We will share the completely annotated analysis correlating clinical metrics to the spectrum of epigenetic mutations and GDMS and across all histology’s and treatments. Clinical trial information: NCT01998035; NCT01947140.

Introduction:Hunger is a global problem and has increased in recent years. In Latin America, hunger continues in high numbers. Although the level of hunger is relatively low compared to other regions, this increase in Latin America is mainly explained by the economic slowdown in South America. Also, climate changes are already weakening the production of the main crops in tropical and temperate regions.Aim:Report the numbers of hunger in Latin America.Methods:A cross-sectional study with reports of the World Health Organization’s hunger figures, September 2018.Results:The number of hungry people in the world has increased for the third consecutive year and affects 821 million people, according to a report released by UN agencies. This corresponds to one in nine people in the world. In Brazil, the figures indicate that more than 5.2 million people spent a day or more without consuming food by 2017, which corresponds to 2.5% of the population. In Latin America and the Caribbean, hunger has also increased and affects some 39 million people.Discussion:Hunger is a catastrophic problem in Latin America. Involving professionals in food and nutrition to try to reduce these numbers appears to be a good strategy because just as the doctor treats the disease, the involvement of other specialists to address the cause of the problem can bring long-term benefits. A social project for this purpose that mobilizes chefs and nutritionists is in progress in Brazil.

The complex interplay between cytokines and chemokines regulates innate and adaptive immune responses against pathogens; specifically, cytokine and chemokine expression drives activation of immune effector cells and their recruitment to tissue infection sites. Herein, we inoculated dogs with Leishmania braziliensis antigens plus saponin (the LBSap vaccine), as well as with the vaccine components, and then used real-time PCR to evaluate the kinetics of dermal expression of mRNAs of cytokines (IL-12, IFN-γ, TNF-α, IL-4, IL-13, TGF-β and IL-10) and chemokines (CCL2, CCL4, CCL5, CCL21 and CXCL8) 1, 12, 24 and 48 h after inoculation. We also evaluated the correlation between cytokine and chemokine expression and dermal cellularity. The LBSap vaccine induced high levels of IL-12 and IL-10 expression at 12 and 24 h, respectively. Furthermore, we observed positive correlations between IL-12 and IL-13 expression, IFN-γ and IL-13 expression, and IL-13 and TGF-β expression, suggesting that a mixed cytokine microenvironment developed after immunization with the vaccine. Inoculation with the saponin adjuvant alone induced a chemokine and cytokine expression profile similar to that observed in the LBSap group. CCL4 and CXCL8 chemokine expression was up regulated by the LBSap vaccine. CCL5 expression was initially highest in the LBSap group, but at 48 h, expression was highest in the LB group. Information about the kinetics of the immune response to this vaccine gained using this dog model will help to elucidate the mechanisms of and factors involved in a protective response against Leishmania infection and will aid in establishing rational approaches for the development of vaccines against canine visceral leishmaniasis.

Phytases of different origin were covalently bound onto Fe3O4 magnetic nanoparticles (12 nm). Binding efficiencies of all three phytases were well above 70% relative to the number of aldehyde groups available on the surface of the magnetic nanoparticles. Temperature stability for all three phytases was enhanced as a consequence of immobilisation, whereas pH dependence of enzyme activity was not affected. Maximum catalytic activity of the immobilised phytases was found at 60°C (rye), 65°C (Aspergillus niger) and 70°C (Escherichia albertii). The immobilised enzymes exhibited the same excellent substrate specificities and unique myo-inositol phosphate phosphatase activities as their soluble counterparts. However, the catalytic turnover number dropped drastically for the immobilised phytases. The amount of the desired partially phosphorylated myo-inositol phosphate isomer could be easily controlled by the contact time of substrate solution and immobilised enzymes. The immobilised phytases showed a high operational stability by retaining almost full activity even after fifty uses.

The role of interleukin IL-4, IL-10 and interferon gamma cytokines on natural Fasciola hepatica infection was investigated by quantifying the mRNA levels in liver tissue from chronically infected cattle. IL-4 and IL-10 had higher expression relative to interferon gamma in the liver tissue of infected animals when compared with the control group. The higher levels of IL-10 and IL-4 observed in the present study suggest a synergism between these cytokines, as well as involvement in the suppression of TH1 cell responses and a consequent induction of decreased interferon gamma expression in chronic cattle fascioliasis. The cytokine ratios were positively correlated, indicating a predominance of IL-4 in the chronic phase of infection with respect to interferon gamma and IL-10. Interferon gamma was predominant expressed in the controls, suggesting the involvement of IL-10 in modulating the immune response in favor of IL-4 in infected animals. Our results suggest that the TH2 polarized host immune response previously observed in experimental infection may also be responsible for establishing chronic phase and the maintenance of the natural infection of cattle from endemic areas that are in continuous contact with parasite.

We have used an integrated approach to study the mobility of inorganic phosphorus (P) from soil solid phase as well as the microbial biomass P and respiration at increasing doses of citric and oxalic acid in two different soils with contrasting agronomic P status. Citric or oxalic acids significantly increased soil solution P concentrations for doses over 2 mmol kg

Tegumentary leishmaniasis (TL) constitutes a major public health problem with significant morbidity worldwide. Synthetic peptide-based vaccines are attractive candidates to protect against leishmaniasis, since T cell-specific epitopes can be delivery to antigen-presenting cells, leading to the generation of a Th1 cell-mediated immunity. In this context, the present study aims to evaluate the immunogenicity and protective efficacy of a vaccine composed of major histocompatibility complex class I and II-restricted epitopes derived from four Leishmania infantum proteins to protect mice against Leishmania amazonensis infection. This recombinant fusion protein was administered in BALB/c mice alone or with saponin. As controls, animals received saline or saponin. In the results, the administration of the recombinant protein plus saponin induced a specific IFN-γ, IL-12 and GM-CSF production, as well as high IgG2a isotype antibody levels, which protected mice against a challenge using L. amazonensis promastigotes. Lower parasite burden was found in the infected footpads, liver, spleen and draining lymph node of vaccinated mice, when compared to those from the control groups. In addition, protection was associated with a lower IL-4 and IL-10 response, which was accompanied by the antileishmanial nitrite production by spleen cells of the animals. Interestingly, the recombinant protein administered alone induced a partial protection against challenge. In conclusion, this study shows a new vaccine candidate based on T cell-specific epitopes that was able to induce protection against L. amazonensis infection.

In the present study, two Leishmania braziliensis proteins, one hypothetical and the eukaryotic initiation factor 5a (EiF5a), were cloned and used as a polyproteins vaccine for the heterologous protection of BALB/c mice against infantum infection. Animals were immunized with the antigens separately or in association, and in both cases saponin was used as an adjuvant. In the results, spleen cells from mice inoculated with the individual or polyproteins vaccine and lately challenged produced significantly higher levels of protein- and parasite-specific IFN-γ, IL-12, and GM-CSF, when both a capture ELISA and flow cytometry assays were performed. Evaluating the parasite load by a limiting dilution as well as by RT-PCR, these animals presented significant reductions in the parasite number in all evaluated organs, when compared to the control (saline and saponin) groups. The best protection was reached when the polyproteins vaccine was employed. Protection was associated with the IFN-γ production against parasite extracts, which was mediated by both CD4(+) and CD8(+) T cells and correlated with the antileishmanial nitrite production. In this context, this vaccine combining two L. braziliensis proteins was able to induce a heterologous protection against VL, and could be considered in future studies to be tested against other Leishmania species or in other mammalian hosts.

Resumo No Seminário da ética, Lacan opôs a pastoral analítica à ética do “não ceder de seu desejo”. Ao pôr o desejo como parâmetro, tal ética questiona valores universais para alcançar uma suposta boa vida. Comparamos tal formulação à ética do “bem-dizer o desejo”, destacada por Lacan em Televisão. Se no Seminário 7 temos uma crítica aos ideais de felicidade associados ao mercado dos bens, na ética do “bem-dizer o desejo”, a felicidade aparece como parte do sintoma, vinculada aos diferentes modos de gozo, via possível para questionar a atual injunção a aproveitar, vinculada a uma certa cultura da alta performance. Abstract In the Seminar of ethics, Lacan has opposed the analytical pastoral and the “do not give away on your desire” ethics. By putting the desire as a parameter, such an ethics questions the universal values to achieve a supposed good life. Here we compare this formulation to the ethics of the “benedictory of desire”, highlighted by Lacan in Television. If in the 7th Seminar we have a critics about the ideal of happiness, associated to the market of the goods, on the “benedictoty of desire” ethics happiness appears only as a symptom, tied to the different modes of enjoyment, which is a possible way to question about the contemporary injunction to enjoy, bound to a certain “high performance” culture.

The current treatment of leishmaniasis has been hampered due to the high toxicity of the available drugs and long duration protocols, which often lead to its abandonment. In the present study, a poloxamer 407-based delivery system was developed, and a molecule, 8-hydroxyquinoline (8-HQN), was incorporated with it, leading to an 8-HQN/micelle (8-HQN/M) composition. Assays were performed to evaluate the in vitro antileishmanial activity of 8-HQN/M against Leishmania amazonensis stationary promastigotes. The cytotoxicity in murine macrophages and in human red cells, as well as the efficacy of the treatment in macrophages infected by parasites, was also assessed. This product was also evaluated for the treatment of murine tegumentary leishmaniasis, using L. amazonensis-infected BALB/c mice. To evaluate the in vivo efficacy of the treatment, the average lesion diameter (area) in the infected tissue, as well as the parasite load at the site of infection (skin), spleen, liver and draining lymph nodes were examined. Non-incorporated micelle (B-8-HQN/M) and the free molecule (8-HQN) were used as controls, besides animals that received only saline. The parasite burden was evaluated by limiting dilution and quantitative real-time PCR (qPCR) techniques, and immunological parameters associated with the treatments were also investigated. In the results, the 8-HQN/M group, when compared to the others, presented more significant reductions in the average lesion diameter and in the parasite burden in the skin and all evaluated organs. These animals also showed significantly higher levels of parasite-specific IFN-γ, IL-12, and GM-CSF, associated with low levels of IL-4 and IL-10, when compared to the saline, 8-HQN/M, and B-8-HQN groups. A predominant IL-12-driven IFN-γ production, against parasite proteins, mainly produced by CD4+ T cells, was observed in the treated animals, post-infection. In conclusion, 8-HQN/M was highly effective in treating L. amazonensis-infected BALB/c mice and can be considered alone, or combined with other drugs, as an alternative treatment for tegumentary leishmaniasis. Graphical Abstract Therapeutic scheme and immunological and parasitological parameters developed in the present study.

Organic wastes rich in phosphorus (P) are considered an alternative to decrease the dependence on chemical P fertilization in crops and pastures. Microbial inoculants are being studied as a tool to increase plant P availability in organic wastes. In this study, we explore the effect of inoculation with Bacillus sp. MQH-19 (a native phytase-producing bacterium) on the release of inorganic phosphorus (Pi) in cattle manure with low available P but a high total P content. Bacteria inoculation resulted in a higher release of Pi (8% in NaHCU3 and 13% in NaOH-EDTA extracts) compared with that of uninoculated manure (0.7% in NaHCU3 and 0.1% in NaOH-EDTA extracts). However, a greater amount of Pi was released in inoculated manure supplemented with phytate (47% in NaHCU3 and 117% in NaOH-EDTA extracts) compared with that of uninoculated manure supplemented with phytate (30% in NaHCU3 and 15% in NaOH-EDTA extracts). In addition, the use of denaturing gradient gel electrophoresis (DGGE) and quantitative PCR (qPCR) revealed that the bacterial community structure in manure was affected by inoculation and that the prevalence of Bacillus sp. MQH-19 decreased during incubation (6 days). This study demonstrates that Pi availability in cattle manure can be increased by phytase-producing bacteria inoculation. Phytase-producing bacteria inoculation might represent an attractive strategy to increase P availability in agricultural wastes, which are used as organic fertilizers in crops and pastures.

The mobility and resupply of inorganic phosphorus (P) from the solid phase were studied in 32 soils from the UK. The combined use of diffusive gradients in thin films (DGT), diffusive equilibration in thin films (DET) and the "DGT-induced fluxes in sediments" model (DIFS) were adapted to explore the basic principles of solid-to-solution P desorption kinetics in previously unattainable detail. On average across soil types, the response time (Tc) was 3.6 h, the desorption rate constant (k-1) was 0.0046 h(-1), and the desorption rate was 4.71 nmol l(-1) s(-1). While the relative DGT-induced inorganic P flux responses in the first hour is mainly a function of soil water retention and % Corg, at longer times it is a function of the P resupply from the soil solid phase. Desorption rates and resupply from solid phase were fundamentally influenced by P status as reflected by their high correlation with P concentration in FeO strips, Olsen, NaOH-EDTA and water extracts. Soil pH and particle size distribution showed no significant correlation with the evaluated mobility and resupply parameters. The DGT and DET techniques, along with the DIFS model, were considered accurate and practical tools for studying parameters related to soil P desorption kinetics.

Human tegumentary leishmaniasis (HTL), characterized by skin ulcers that may spread and cause dreadful and massive tissue destruction of the nose and mouth, is considered a neglected tropical disease, and it is a serious threat to global health due to its continuous expansion, favored by the lifecycle of its causative organism that is maintained in domestic animal reservoirs and anthropophilic sand fly species. Serodiagnosis of HTL is a great challenge due to many biological factors, including hampered specificity and/or sensitivity. This investigation addresses the unmet need for new diagnostic markers of HTL, and describes a simple platform to improve the serodiagnosis. A constrained conformational phage display random peptide library combined with a magnetic microsphere-based subtraction strategy was used to identify ligands with potential diagnostic applications. Six clones were selected against IgG antibodies from HTL patients, characterized by sequencing and confirmed by a phage-ELISA using sera from patients developing visceral leishmaniasis (n=20), Chagas disease (n=10), mucosal (n=30) and cutaneous (n=20) leishmaniasis; as well as from healthy subjects living in endemic (n=20) and non-endemic (n=30) areas of leishmaniasis. A wild-type M13-phage clone and a soluble Leishmania antigenic extract were used as negative and positive controls, respectively. Three clones reached 100% sensitivity and specificity, without any cross-reactivity with sera from patients with leishmaniasis-related diseases. Briefly, we describe for the first time a set of serological markers based on three immunodominant mimotopes that showed 100% accuracy, and that could be used in a phage-ELISA assay for the HTL serodiagnosis.

New therapeutics are urgently needed to treat visceral leishmaniasis (VL). Due to the fact that drug discovery is a long and expensive process, the development of delivery systems to carry old and toxic drugs could be considered, as well as the evaluation of new molecules that have already shown to present biological activity. In this context, the present study evaluated the in vitro and in vivo antileishmanial activity of an 8-hydroxyquinoline (8-HQN)-containing polymeric micelle (8-HQN/M) system against Leishmania infantum, the main causative agent of VL in the Americas. The experimental strategy used was based on the evaluation of the parasite load by a limiting-dilution technique in the spleen, liver, bone marrow and draining lymph nodes of the infected and treated animals, as well as by a quantitative PCR (qPCR) technique to also assess the splenic parasite load. The immune response developed was evaluated by the production of IFN-γ, IL-4, IL-10, IL-12 and GM-CSF cytokines, as well as by antileishmanial nitrite dosage and antibodies production. Hepatic and renal enzymes were also investigated to verify cellular injury as a result of treatments toxicity. In the results, 8-HQN/M-treated mice, when compared to the other groups: saline, free amphotericin B (AmpB, as a drug control), 8-HQN and B-8-HQN/M (as a micelle control) showed more significant reductions in their parasite burden in all evaluated organs. These animals also showed an antileishmanial Th1 immunity, which was represented by high levels of IFN-γ, IL-12, GM-CSF and nitrite, associated with a low production of IL-4 and IL-10 and anti-Leishmania IgG1 isotype antibodies. In addition, any hepatic or renal damage was found in these treated animals. In conclusion, 8-HQN/M was effective in treating L. infantum-infected BALB/c mice, and can be considered alone, or combined with other drugs, as an alternative treatment for VL.

All rights reserved. Visceral leishmaniasis (VL) is one of the most important tropical diseases worldwide. Although chemotherapy has been widely used to treat this disease, problems related to the development of parasite resistance and side effects associated with the compounds used have been noted. Hence, alternative approaches for VL control are desirable. Some methods, such as vector control and culling of infected dogs, are insufficiently effective, with the latter not ethically recommended. The development of vaccines to prevent VL is a feasible and desirable measure for disease control, for example, some vaccines designed to protect dogs against VL have recently been brought to market. These vaccines are based on the combination of parasite fractions or recombinant proteins with adjuvants that are able to induce cellular immune responses, however, their partial efficacy and the absence of a vaccine to protect against human leishmaniasis underline the need for characterization of new vaccine candidates. This review presents recent advances in control measures for VL based on vaccine development, describing extensively studied antigens, as well as new antigenic proteins recently identified using immuno-proteomic techniques, This work was supported by grants from Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica, Rede Nanobiotec/Brasil-Universidade Federal de Uberlândia/CAPES, PRONEX-FAPEMIG (APQ-01019-09), FAPEMIG (CBB-APQ-00819-12 and CBB-APQ-01778-2014), and CNPq (APQ-482976/2012-8, APQ-488237/2013-0, and APQ-467640/2014-9). EAFC and LRG are recipients of the grant from CNPq. MACF is the recipient of grants from FAPEMIG/CAPES

O artigo discute as relações entre a psicanálise e a ciência, especialmente no que tange aos seus métodos de investigação e trabalho. Para isso, tomamos como objeto a noção de neutralidade, tal como podemos apreendê-la na clínica da psicanálise, para confrontá-la com o que chamamos de isenção no campo da ciência, que não quer dizer outra coisa que a neutralidade proposta lá. Veremos que, apesar da clara filiação da noção psicanalítica para com a noção vinda da ciência, a neutralidade clínica da psicanálise toma outro rumo, que se pode mesmo opor ao sentido do termo na ciência. Enquanto em ciência o termo quer significar um processo de purificação e de seleção, em psicanálise ele surge como a exigência de nada selecionar. This article discusses the relations between psychoanalysis and science, with special attention on their investigation and working methods. For that, we take as object the notion of neutrality, as we can apprehend on the psychoanalytical clinics, to confront it with what we’ll call exemption on science’s field, that doesn’t mean other than the neutrality proposed there. We’ll see that, despite the clear heritage of the psychoanalytical notion to the one from science, the clinical neutrality of psychoanalysis takes another route, one that can be even opposed to the sense of the term in science. While in science the term means a process of purification and selection, in psychoanalysis it arises as a requirement of nothing select. Cet article discute les rapports entre la psychanalyse et la science, avec une attention particulière par ces méthodes d’investigation. Pour ça, on prends la notion de neutralité, comme on peut lire a partir de la clinique psychanalytique, a fin de la confronter avec ce qu’on peut appeler d’exemption au champ scientifique, et qui ne veut dire autre chose que la neutralité qu’on se propose là. On vas voire que, malgré la claire filiation que de la notion psychanalytique par rapport à la notion scientifique, la neutralité clinique de la psychanalyse prends autre chemin, qui peut être même opposé au sens du terme chez la science. Si, à la science, le terme signifie un procès de purification et sélection, chez la psychanalyse il paraît comme une exigence de rien choisir. El artículo discute las relaciones entre el psicoanálisis y la ciencia, con atención especial para los métodos de investigación y trabajo. Para tal, tomaremos como objeto la noción de neutralidad, como la aprehendemos en la clínica psicoanalítica, para confrontarla con lo que acá llamamos exención en el campo científico, lo que no es otra cosa que la neutralidad propuesta allá. Veremos que, a pesar de la clara filiación que la noción psicoanalítica presenta por relación a la científica, la neutralidad del psicoanálisis toma otro rumo, que puede ser mismo el opósito del sentido científico del término. Mientras que en la ciencia el término significa un proceso de purificación y selección, en psicoanálisis surge como exigencia de nada seleccionar.

O presente artigo retoma a noção de neutralidade proveniente da técnica clínica psicanalítica, confrontando-a com noções próximas, tais como a neutralidade científica e o neutro na semiologia de Barthes. Procuramos demonstrar como tal noção descreve bem a postura analítica, exigida tanto do paciente (associação livre) quanto do analista (atenção flutuante), e como ela nasce na prática clínica freudiana, respondendo a problemas decorrentes dela. Assim, abordaremos os temas da hipnose e da sugestão, como também aqueles ligados à contra-transferência. Defenderemos, ainda, que a neutralidade não é apenas questão de técnica, mas é central para o entendimento da própria ética da psicanálise.

Following the discovery of NVP-BEZ235, our first dual pan-PI3K/mTOR clinical compound, we sought to identify additional phosphoinositide 3-kinase (PI3K) inhibitors from different chemical classes with a different selectivity profile. The key to achieve these objectives was to couple a structure-based design approach with intensive pharmacologic evaluation of selected compounds during the medicinal chemistry optimization process. Here, we report on the biologic characterization of the 2-morpholino pyrimidine derivative pan-PI3K inhibitor NVP-BKM120. This compound inhibits all four class I PI3K isoforms in biochemical assays with at least 50-fold selectivity against other protein kinases. The compound is also active against the most common somatic PI3Kα mutations but does not significantly inhibit the related class III (Vps34) and class IV (mTOR, DNA-PK) PI3K kinases. Consistent with its mechanism of action, NVP-BKM120 decreases the cellular levels of p-Akt in mechanistic models and relevant tumor cell lines, as well as downstream effectors in a concentration-dependent and pathway-specific manner. Tested in a panel of 353 cell lines, NVP-BKM120 exhibited preferential inhibition of tumor cells bearing PIK3CA mutations, in contrast to either KRAS or PTEN mutant models. NVP-BKM120 shows dose-dependent in vivo pharmacodynamic activity as measured by significant inhibition of p-Akt and tumor growth inhibition in mechanistic xenograft models. NVP-BKM120 behaves synergistically when combined with either targeted agents such as MEK or HER2 inhibitors or with cytotoxic agents such as docetaxel or temozolomide. The pharmacological, biologic, and preclinical safety profile of NVP-BKM120 supports its clinical development and the compound is undergoing phase II clinical trials in patients with cancer. Mol Cancer Ther; 11(2); 317–28. ©2011 AACR.

Phosphoinositide-3-kinases (PI3Ks) are important oncology targets due to the deregulation of this signaling pathway in a wide variety of human cancers. Herein we describe the structure guided optimization of a series of 2-morpholino, 4-substituted, 6-heterocyclic pyrimidines where the pharmacokinetic properties were improved by modulating the electronics of the 6-position heterocycle, and the overall druglike properties were fine-tuned further by modification of the 4-position substituent. The resulting 2,4-bismorpholino 6-heterocyclic pyrimidines are potent class I PI3K inhibitors showing mechanism modulation in PI3K dependent cell lines and in vivo efficacy in tumor xenograft models with PI3K pathway deregulation (A2780 ovarian and U87MG glioma). These efforts culminated in the discovery of 15 (NVP-BKM120), currently in Phase II clinical trials for the treatment of cancer.

Os aneurismas da artéria radial são raros e esporádicos, sendo que os pseudoaneurismas são mais frequentes do que os aneurismas verdadeiros e maioritariamente de configuração sacular. A etiologia é variada e diferenciam-se de outros diagnósticos pela pulsação e frémito. Raramente se complicam de ruptura, sendo a trombose e embolização as principais complicações. Dos casos publicados sobre aneurismas verdadeiros da artéria radial, apenas um está descrito como sendo secundário a lesão ocupacional repetitiva, sendo a maioria de causa idiopática. Os autores descrevem um caso de uma mulher de 63 anos, referenciada à consulta de Cirurgia Vascular por crescimento de massa pulsátil na tabaqueira anatómica da mão esquerda. O estudo por eco-doppler e angiografia, confirmaram o diagnóstico de aneurisma sacular da artéria radial, com 20 mm de maior eixo, arcada palmar permeável e sem sinais de embolização distal. Foi submetida a aneurismectomia parcial com laqueação dupla proximal e distal e endoaneurismorrafia. A cirurgia e pós-operatório decorreram sem complicações, nomeadamente complicações isquémicas. A propósito desde caso clínico, discute-se a abordagem diagnóstica e opções terapêuticas.Radial artery aneurysms are sporadic and rare, pseudoaneurysms are more common than true aneurysms, mainly in saccular configuration. The etiology is varied and difference from other diagnostics is done by the presence of pulse and thrill. Thrombosis and embolization are the main complications, while rupture is rare. From the reported cases of true aneurysms of the radial artery, only one is described as being secondary to repetitive occupational injury, the majority being idiopathic. The authors describe the case of a 63 year old woman, referred to a Vascular Surgery consultation because of a growing pulsatile mass in the anatomical snuffbox of the left hand. The Doppler and Angiography studies confirmed the diagnosis of saccular aneurysm of the radial artery, 20 mm on the long axis, with permeable palmar arch and without signs of distal embolization. She underwent a partial aneurysmectomy with ligation and double proximal and distal endoaneurysmorrhaphy. There were no surgical or postoperative complications, particularly ischemic complications. Regarding this case, diagnosis and treatment options are discussed.

Trata-se de apresentar a contribuição de Paul Feyerabend ao debate entre psicanálise e ciência. A obra deste autor parece fornecer um novo horizonte de discussão, pois esvazia a pretensão de idealizar a ciência e a apresenta como atividade humana, impura, sem qualquer regra racional de funcionamento. Primeiro, faremos uma 'varredura' pelo campo atual de discussão, destacando alguns artigos que contribuem para esse debate. Seguiremos com uma crítica ao esforço de regramento epistemológico; e, por fim, apresentaremos a obra de Feyerabend, concentrando nossa atenção na leitura do célebre volume Contra o método, e no que esta obra acrescenta ao debate.Our goal is to present the contribution of Paul Feyerabend's works to the debate between psychoanalysis and science. Feyerabend's works seem to presents a new discussion horizon, since it empties out there the pretension to idealize science, and presents it as a human activity, impure, without any rational rule of functioning. First, we will scan the current discussion field, highlighting some articles that contribute to the debate. We will follow by doing some critics to epistemology and its ruling efforts. Finally, we will present Feyerabend's works, concentrating our attention on the lecture of Against Method, his famous volume, and what this work adds to the debate.

Phytate is one of the most abundant sources of organic phosphorus (P) in soils, but must be mineralized by phytase-producing bacteria to release P for plant uptake. Microbial inoculants based on Bacillus spp. have been developed commercially, but few studies have evaluated the ecology of these bacteria in the rhizosphere or the types of enzymes that they produce. Here, we studied the diversity of aerobic endospore-forming bacteria (EFB) with the ability to mineralize phytate in the rhizosphere of pasture plants grown in volcanic soils of southern Chile. PCR methods were used to detect candidate phytase-encoding genes and to identify EFB bacteria that carry these genes. This study revealed that the phytate-degrading EFB populations of pasture plants included species of Paenibacillus and Bacillus, which carried genes encoding β-propeller phytase (BPP). Assays of enzymatic activity confirmed the ability of these rhizosphere isolates to degrade phytate. The phytase-encoding genes described here may prove valuable as molecular markers to evaluate the role of EFB in organic P mobilization in the rhizosphere.

Introduction: Angiogenesis plays an important role in the malignancy of glioblastoma (GBM) as well as in cancer immunotherapy. In addition, PD-1 checkpoint receptors are upregulated in GBM. We investigate the treatment effects of combination immunotherapy with checkpoint inhibitor anti-PD-1 and anti-CXCR4, an antagonist of a chemokine receptor involved in immune cell homing as well as vasculature development, in a murine glioma model. Methods: C57Bl/6 mice were implanted with GL261-Luc+ glioma cells and randomized into 4 treatment arms: 1) control, 2) anti-PD-1 monotherapy, 3) anti-CXCR4 monotherapy, and 4) combination anti-PD-1 and anti-CXCR4 therapy. Overall survival and median survival were assessed. Brain samples from untreated mice were stained for CXCR4 expression on plasma membrane, and immunohistochemistry studies were conducted on human glioblastoma specimens as well. Immune cell activation and cell population changes were assessed through flow cytometry analysis. Results: Both murine and human glioma specimens demonstrated robust positive expression of CXCR4 on tumor-infiltrating immune cells and endothelial cells of vasculature specific to the tumor bed. Combination therapy with anti-PD-1 and anti-CXCR4 conferred survival benefit compared to control and both monotherapy arms. Long-term survivors that had received combination therapy demonstrate immune memory and decreased populations of immunosuppressive, tumor-promoting immune cells. For instance, the monocytic myeloid-derived suppressor cell population was decreased in the brain in mice treated with combination therapy. The pattern of change in microglia was similar in the brain compartment for the combination treatment group as well. Conclusion: Anti-CXCR4 and anti-PD-1 synergistic immunotherapy not only modulates the immune cell make-up of the glioma microenvironment but also affects vasculature within the tumor region. Dual therapy targeting both checkpoint and chemokine receptors results in improved survival rates. Citation Format: Adela Wu, Pina Cardarelli, Miho Oyasu, Daniel Menezes, Paul Ponath, John Cogswell, Russell Maxwell, Andrew Luksik, Alice Hung, Eileen Kim, Zineb Belcaid, Henry Brem, Drew Pardoll, Michael Lim. The combination of CXCR4 and checkpoint receptor inhibition improves survival in an orthotopic murine glioma model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1736.

The inhibition of key receptor tyrosine kinases (RTKs) that are implicated in tumor vasculature formation and maintenance, as well as tumor progression and metastasis, has been a major focus in oncology research over the last several years. Many potent small molecule inhibitors of vascular endothelial growth factor receptor (VEGFR) and platelet-derived growth factor receptor (PDGFR) kinases have been evaluated. More recently, compounds that act through the complex inhibition of multiple kinase targets have been reported and may exhibit improved clinical efficacy. We report herein a series of potent, orally efficacious 4-amino-3-benzimidazol-2-ylhydroquinolin-2-one analogues as inhibitors of VEGF, PDGF, and fibroblast growth factor (FGF) receptor tyrosine kinases. Compounds in this class, such as 5 (TKI258), are reversible ATP-competitive inhibitors of VEGFR-2, FGFR-1, and PDGFRbeta with IC(50) values

O arroz é um dos alimentos mais consumidos no mundo, caracterizando-se como a base da alimentação para mais da metade da população mundial. De acordo com a Organização Mundial da Saúde (WHO), a ingestão de alimentos contaminados é uma das principais vias de absorção de elementos potencialmente tóxicos pelo ser humano. Por esse motivo, órgãos de fiscalização têm estabelecidos limites máximos permitidos desse tipo de elemento nos alimentos. Nesse sentido, o desenvolvimento de métodos e aplicações visando à determinação de elementos com caráter tóxico, tais como o Al, As, Cd e Pb em alimentos é de suma importância. Nesse projeto, foram desenvolvidos métodos para determinações diretas de Al, As, Cd e Pb em arroz por espectrometria de absorção atômica com forno de grafite e amostragem direta de sólidos (SS-GF AAS) que tem se mostrado uma técnica promissora especialmente devido à sua elevada sensibilidade e detectabilidade, além de dispensar morosos processos de decomposição de amostra. A adequada otimização do programa de aquecimento aliado ao uso de modificadores químicos adequados possibilitou o uso de calibração aquosa. Parâmetros importantes como o tamanho de partícula, massa de amostra e segregação de analito foram investigados. Observou-se que partículas inferiores a 100 µm geravam resultados precisos. A massa mínima ideal de amostra de arroz a ser utilizada na análise foi em torno de 300 µg. A massa máxima recomendada foi 500 µg. Segregação de Al e Cd não foram observadas na amostra de arroz. A segunda parte do projeto envolveu a aplicação do método de análise desenvolvido em amostras de arroz comercializadas na cidade de São Paulo. Investigou-se também a influência dos processos de lavagem e cozimento dos grãos sobre a concentração de Al, As, Cd e Pb. Todas as 17 amostras de diversos tipos e marcas de arroz analisadas apresentaram concentrações de As, Cd e Pb abaixo dos limites estabelecidos por lei. Com relação ao Al, não há regulamentado o valor de limite máximo. As maiores concentrações desse elemento foram observadas nos grãos de arroz integral. O processo de lavagem removeu significativamente a concentração de Al e Pb na maioria das amostras, entretanto, o mesmo não foi observado para o Cd. O processo de cozimento diminuiu a concentração de Cd e Pb em parte das amostras, e um pequeno aumento na concentração do Al foi observado, o qual pode ser atribuído à contaminação pelo ambiente. O cozimento do arroz em frascos de vidro ou panelas a base de alumínio não alterou a concentração desse elemento no arroz, nem mesmo quando a panela de alumínio foi polida com palha de aço Rice is a staple food for more than 40% of the world\'s population. Considering that the main reason of contamination by potentially toxic element occurs by food ingestion, World Health Organization (WHO) has been controlling the levels of some contaminants. In this context, development of methods and applications associated to determination of potentially toxic elements such as Al, As, Cd and Pb in food are important. In this work, methods for direct determination of Al, As, Cd and Pb in rice by solid sampling graphite furnace atomic absorption spectrometry (SS-GF AAS) were developed. Solid sampling-GF AAS has shown as a promising technique especially due to its high sensitivity and detectability and the possibility to practice direct analysis of solid samples, dismissing tedious and error prone sample preparation step. The adequate heating program optimization associated to the use of chemical modifier allowed direct rice analysis against aqueous calibration. The influence of some parameters, such as particle size, sample mass size and analyte segregation on the analytical results were investigated. Results showed that particle size smaller than 100 µm generated accurate results. The minimum sample mass size recommended for analysis was around 300 µg and the maximum sample mass size was around 500 µg. Segregation of Cd and Al was not observed in the rice sample. The second part of the work involved the analysis of 17 rice samples commercially available in São Paulo city. Influence of rice washing and cooking procedure on the Al, As, Cd and Pb concentrations was also investigated. Concentration of all analytes were lower than the maximum allowed by WHO in all samples. The highest concentrations of Al were found in coarse rice. The washing process reduced significantly the concentrations of Al and Pb in most samples, however, the same was not observed for Cd. Cooking process reduced the concentration of Cd and Pb for most samples, probably by volatilization. On the other hand, an increase in Al concentration was observed. This fact could be attributed to the sample contamination during cooking process. Rice cooked in glass or aluminum container presented the same concentration of Al, even when polished aluminum container was used. These results indicated that there is no translocation of Al from container to food

The serodiagnosis of human tegumentary leishmaniasis (TL) presents some problems, such as the low level of antileishmanial antibodies found in most of the patients, as well as the cross-reactivity in subjects infected by other trypanosomatids. In the present study, an immunoproteomic approach was performed aimed at identification of antigens in total extracts of stationaryphase promastigote and amastigote-like forms of Leishmania (Viannia) braziliensis using sera from TL patients. With the purpose of reducing the cross-reactivity of the identified proteins, spots recognized by sera from TL patients, as well as those recognized by antibodies present in sera from noninfected patients living in areas where TL is endemic and sera from Chagas disease patients, were discarded. Two Leishmania hypothetical proteins and 18 proteins with known functions were identified as antigenic. The study was extended with some of them to validate the results of the immunoscreening. The coding regions of five of the characterized antigens (enolase, tryparedoxin peroxidase, eukaryotic initiation factor 5a, β-tubulin, and one of the hypothetical proteins) were cloned in a prokaryotic expression vector, and the corresponding recombinant proteins were purified and evaluated for the serodiagnosis of TL. The antigens presented sensitivity and specificity values ranging from 95.4 to 100% and 82.5 to 100%, respectively. As a comparative antigen, a preparation of Leishmania extract showed sensitivity and specificity values of 65.1 and 57.5%, respectively. The present study has enabled the identification of proteins able to be employed for the serodiagnosis of TL., Instituto Nacional de Ciência e Tecnologia em Nano-biofarmacêutica (INCT-Nanobiofar), FAPEMIG (CBB-APQ-0496-11 and CBB-APQ-00819-12), and CNPq (APQ-472090/2011-9, APQ-482976/2012-8, and APQ-488237/2013-0). In addition, this study was partially funded in Madrid by a Spanish grant from Ministerio de Economía y Competitividad-FEDER (FISPI14/00366 from the Instituto de Salud Carlos III)

The development of new therapeutic strategies to treat leishmaniasis has become a priority. In the present study, the antileishmanial activity of 8-hydroxyquinoline (8-HQN) was investigated against in vitro promastigotes and in vivo intra-macrophage amastigotes of three Leishmania species: Leishmania amazonensis, Leishmania infantum and Leishmania braziliensis. Studies were performed to establish the 50% Leishmania inhibitory concentration (IC50) of 8-HQN, as well as its 50% cytotoxic concentration (CC50) on murine macrophages and in human red blood cells. The inhibition of macrophages infection was also evaluated using parasites that were pre-treated with 8-HQN. The effects of this compound on nitric oxide (NO) production and in the mitochondrial membrane potential were also evaluated. Finally, the therapeutic efficacy of 8-HQN was assessed in a known murine model, L. amazonensis-chronically infected BALB/c mice. Our results showed that 8-HQN was effective against promastigote and amastigote stages of all tested Leishmania species, presenting a selectivity index of 328.0, 62.0 and 47.0 for L. amazonensis, L. infantum and L. braziliensis, respectively. It was effective in treating infected macrophages, as well as in preventing the infection of these cells using pre-treated parasites. In addition, 8-HQN caused an alteration in the mitochondrial membrane potential of the parasites. When administered at 10mg/kg body weight/day by subcutaneous route, this product was effective in reducing the lesion diameter, as well as the parasite load in evaluated tissues and organs of infected animals. The results showed the in vitro and in vivo efficacy of 8-HQN against three different Leishmania species causing tegumentary and/or visceral leishmaniasis, and it could well be used for future therapeutic optimization studies to treat leishmaniasis.

The serodiagnosis of canine visceral leishmaniasis (CVL) presents problems related to its sensitivity and/or specificity. In the present study, a new Leishmania-specific hypothetical protein, LiHyD, was produced as a recombinant protein (rLiHyD) and evaluated in ELISA experiments for the CVL serodiagnosis. LiHyD was characterized as antigenic in a recent immunoproteomic search performed with Leishmania infantum proteins and the sera of dogs developing visceral leishmaniasis (VL). Aiming to compare the efficacy between whole proteins and synthetic peptides, two linear and one conformational B cell epitopes of LiHyD were synthesized and also evaluated as diagnostic markers. The four antigens were recognized by the sera of dogs suffering VL. On the contrary, low reactivity was observed when they were assayed with sera from non-infected healthy dogs living in endemic or non-endemic areas of leishmaniasis. In addition, no reactivity was found against them using sera from dogs experimentally infected by Trypanosoma cruzi, Babesia canis, or Ehrlichia canis, or sera from animals vaccinated with the Leish-Tec® vaccine, a prophylactic preparation commercially available for CVL prevention in Brazil. As comparative diagnostic tools, a recombinant version of the amastigote-specific A2 protein and a soluble crude Leishmania extract were studied. Both antigens presented lower sensitivity and/or specificity values than the LiHyD-based products. The rLiHyD presented better results for the CVL serodiagnosis than its linear epitopes, although the peptide recreating the conformational epitope resulted also appropriate as a diagnostic marker of CVL. To the best of our knowledge, this is the first study showing the use of a conformational epitope derived from a Leishmania protein for serodiagnosis of CVL.

The present study aimed to evaluate a new Leishmania-specific hypothetical protein, LiHyT, as a vaccine candidate against VL. The immunogenicity of the recombinant protein (rLiHyT) plus saponin was evaluated in BALB/c mice. In the results, it is shown that rLiHyT plus saponin vaccinated mice produced high levels of IFN-γ, IL-12, and GM-CSF after in vitro stimulation of spleen cells using both rLiHyT and Leishmania infantum SLA. The protective efficacy was evaluated after subcutaneous challenge with stationary promastigotes of L. infantum. Immunized and infected mice, when compared to the controls, showed significant reductions in the number of parasites in the liver, spleen, bone marrow, and in the paws' draining lymph nodes. Protection was associated with an IL-12-dependent production of IFN-γ, mainly by CD4(+) T cells, with a minor contribution of CD8(+) T cells. In these mice, a decrease in the parasite-mediated IL-4 and IL-10 responses, as well as a predominance of LiHyT- and parasite-specific IgG2a isotype antibodies, were also observed. The present study showed that a new Leishmania-specific protein, when combined with a Th1-type adjuvant, presents potential to be used as a vaccine against VL.

Serological diagnostic tests for canine and human leishmaniasis present problems related with their sensitivity and/or specificity. Recently, an immunoproteomic approach performed with Leishmania infantum proteins identified new parasite antigens. In the present study, the diagnostic properties of two of these proteins, cytochrome c oxidase and IgE-dependent histamine-releasing factor, were evaluated for the serodiagnosis of canine visceral (CVL) and human tegumentary (HTL) leishmaniasis. For the CVL diagnosis, sera samples from non-infected dogs living in an endemic or non-endemic area of leishmaniasis, sera from asymptomatic or symptomatic visceral leishmaniasis (VL) dogs, from Leish-Tec ® -vaccinated dogs, and sera from animals experimentally infected by Trypanosoma cruzi or Ehrlichia canis were used. For the HTL diagnosis, sera from non-infected subjects living in an endemic area of leishmaniasis, sera from active cutaneous or mucosal leishmaniasis patients, as well as those from T. cruzi -infected patients were employed. ELISA assays using the recombinant proteins showed both sensitivity and specificity values of 100% for the serodiagnosis of both forms of disease, with high positive and negative predictive values, showing better diagnostic properties than the parasite recombinant A2 protein or a soluble Leishmania antigen extract. In this context, the two new recombinant proteins could be considered to be used in the serodiagnosis of CVL and HTL.

Clínica & Culturav. IV, n. IEditorial

The aim of this study was to compare the biochemical properties of seven commercially available phytase products as well as their catalytic performance in an in vitro simulation of the digestive tract of poultry. Their enzymatic properties relevant with respect to phytate dephosphorylation in the digestive tract of birds were determined under identical assay conditions. All phytase products included in the study showed an acid pH optimum of activity and were capable of releasing the organically bound phosphate from phytate during the in vitro simulation. However, their overall biochemical properties and relative catalytic performances were remarkably different. The in vitro degradation system was considered as a simple and useful tool to evaluate the suitability of a phytase to be used as a feed supplement. Although relevant factors such as dietary P levels, intestinal phytase, and P absorption are not implemented in the system, this approach might help to reduce the number of feeding trials necessary in the search for a better suited phytase for animal feeding application.

Feed is the foremost and highest expense in large-scale breeding of monogastric animals, accounting for more than 70% of the total production costs. For more than 30 years, various enzymes have been used to improve the efficiency of feedstuff utilization. These enzymes are applied to simultaneously address many different practical problems in feed use, such as reducing the presence of antinutritional factors, increasing the digestibility of feed constituents, reducing the viscosity in the digestive tract, allowing the use of low-cost ingredients, and reducing environmental risks related to manure and waste disposal. The main feed enzymes in the market are phytases, xylanases and β-glucanases (cellulases). Nevertheless, other enzymes, such as mannanases, α-galactosidases, pectinases, amylases and proteases, are increasing in use. Poultry and swine sectors are the main feed enzyme consumers; however, ruminants, aquaculture and pets are projected to be large markets in the near future. Recent and future advancements in this field of knowledge are discussed here.

Quantification of metal loading in supported catalysts is imperative; however, the analytical methods are usually time-consuming, expensive, and require sophisticated equipment and sample extraction with hazardous and corrosive strong acids. Herein, we report a new method for quantification of metal in supported catalysts using slurry sampling flame atomic absorption spectrometry. The slurry sampling method was used for determination of gold and the results were similar to those obtained with the traditional extraction method using aqua regia; however, it overcomes the main drawbacks associated with sample digestion by avoiding the use of concentrated toxic reagents, minimizing sample manipulation, and consequently, reducing the risks of contamination and the sample preparation time. Ultrasound radiation optimization associated with the solvent, mass of sample, concentration, and matrix effect in the slurry analysis allowed direct analysis by using the calibration in aqueous medium. The slurry sampling procedure could be applied to other catalysts with different supports and in bimetallic systems.

Deferasirox (DFX), an orally active and clinically approved iron chelator, is being used extensively for the treatment of iron overload. However, its water insolubility makes it cumbersome for practical use. In addition to this, the low efficacy of DFX to remove brain iron prompted us to synthesize and evaluate a DFX-TAT(47-57) peptide conjugate for its iron chelation properties and permeability across RBE4 cell line, an in vitro model of the blood-brain barrier. The water-soluble conjugate was able to remove labile iron from buffered solution as well as from iron overloaded sera, and the permeability of DFX-TAT(47-57) conjugate into RBE4 cells was not affected compared to parent deferasirox. The iron bound conjugate was also able to translocate through the cell membrane.

In this work, the effect of vaccination of a newly described Leishmania infantum antigenic protein has been studied in BALB/c mice infected with this parasite species. The LiHyD protein was characterized after a proteomic screening performed with the sera from dogs suffering visceral leishmaniasis (VL). Its recombinant version was expressed, purified and administered to BALB/c mice in combination with saponin. As a result of vaccination and 10 weeks after challenge using an infective dose of L. infantum stationary promastigotes, vaccinated mice showed lower parasite burdens in different organs (liver, spleen, bone marrow and footpads' draining lymph nodes) than mice inoculated with the adjuvant alone or the vaccine diluent. Protected mice showed anti-Leishmania IgG2a antibodies and a predominant IL-12-driven IFN-γ production (mainly produced by CD4 T cells) against parasite proteins, whereas unprotected controls showed anti-Leishmania IgG1 antibodies and parasite-mediated IL-4 and IL-10 responses. Vaccinated mice showed an anti-LiHyD IgG2a humoral response, and their spleen cells were able to secrete LiHyD-specific IFN-γ, IL-12 and GM-CSF cytokines before and after infection. The protection was correlated with the Leishmania-specific production on nitric oxide. Altogether, the results indicate that the new LiHyD protein could be considered in vaccine formulations against VL., Instituto Nacional de Ci^encia e Tecnologia em Nano-biofarmac^eutica (INCT-NanoBiofar), FAPEMIG (CBB-APQ-00819-12 and CBB-APQ-01778-2014) and CNPq (APQ-482976/2012-8, APQ-488237/2013-0 and APQ-467640/2014-9). In addition, this study was partially funded by the Spanish grant from Ministerio de Economía y Competitividad-FEDER (FIS PI14/00366 from the Instituto de Salud Carlos III)

The calibration and quantitative measurement is the “ Achilles heel ” of the LIBS technique. This paper deals with a method developed for the direct measurement of K and Mg in plant samples. Instrumental parameters were optimized and the best condition found was a 50 μm spot size, 10 Hz laser repetition rate, 75 accumulated laser pulses with 25 mJ/pulse and 0.25 μs of delay time. For method calibration, the use of synthetic standard calibrating material prepared by the addition of increasing concentrations of K and Mg in wood, filter paper and babassu mesocarp was proposed in order to assess the feasibility of using these various matrices in plant samples analysis. The limits of detection of proposed method were 2–30 and 6–27 μg g −1 for K and Mg, respectively. The use of the carbon emission wavelength at 247.856 nm was used as internal standard to improve the analytical results. Certified reference materials of plants were used to check the accuracy of the proposed method and recovery around 82% and 100% were obtained in all cases.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) A method for the multielemental determination of metals and nonmetals in goji berries (Lycium barbarum L.) by axially and radially viewed inductively coupled plasma-optical emission spectrometry (ICP-OES) is presented. The accuracy of the entire proposed method was confirmed by three certified reference material analyses, and the certified values showed a good agreement at a 95% confidence limit (Student's Rest). Univ Fed Sao Paulo, Sao Paulo, Brazil Univ Fed Sao Paulo, Inst Quim, Sao Paulo, Brazil Univ Fed Sao Paulo, Sao Paulo, Brazil Univ Fed Sao Paulo, Inst Quim, Sao Paulo, Brazil FAPESP: 2012/11517-1 FAPESP: 2012/11998-0 CNPq: 475282/2013-2 CNPq: 475455/2013-4 Web of Science

ELISA and RIFI are currently used for serodiagnosis of canine visceral leishmaniasis (CVL). The accuracy of these tests is controversial in endemic areas where canine infections byTrypanosoma cruzimay occur. We evaluated the usefulness of synthetic peptides that were selected through phage display technique in the serodiagnosis of CVL. Peptides were chosen based on their ability to bind to IgGs purified from infected dogs pooled sera. We selected three phage clones that reacted only with those IgGs. Peptides were synthesized, polymerized with glutaraldehyde, and used as antigens in ELISA assays. Each individual peptide or a mix of them was reactive with infected dogs serum. The assay was highly sensitive and specific when compared to solubleLeishmaniaantigen that showed cross-reactivity with anti-T. cruziIgGs. Our results demonstrate that phage display technique is useful for selection of peptides that may represent valuable synthetic antigens for an improved serodiagnosis of CVL.

Purpose: To evaluate the therapeutic and biological effects of CHIR-258, an orally bioavailable, potent inhibitor of class III-V receptor tyrosine kinases, in colon cancer models. Experimental Design: The pharmacologic activity of CHIR-258 was characterized by monitoring target modulation as well as by evaluating the antitumor and antiangiogenic effects in human colon xenograft models. Results: CHIR-258 inhibits vascular endothelial growth factor receptor 1/2, fibroblast growth factor receptor 1/3, and platelet-derived growth factor receptor β (PDGFRβ) and shows both antitumor and antiangiogenic activities in vivo. Treatment of KM12L4a human colon cancer cells with CHIR-258 resulted in a dose-dependent inhibition of vascular endothelial growth factor receptor 1 and PDGFRβ phosphorylation and reduction of phosphorylated extracellular signal-regulated kinase (ERK) levels, indicating modulation of target receptors and downstream signaling. In vivo administration of CHIR-258 resulted in significant tumor growth inhibition and tumor regressions, including large, established tumors (500-1,000 mm3). Immunohistochemical analysis showed a reduction of phosphorylated PDGFRβ and phosphorylated ERK in tumor cells after oral dosing with CHIR-258 compared with control tumors. These changes were accompanied by decreased tumor cell proliferation rate and reduced intratumoral microvessel density. CHIR-258 inhibited the phosphorylation of PDGFRβ and ERK phosphorylation in tumors within 2 hours following dosing and the inhibitory activity was sustained for >24 hours. Significant antitumor activity was observed with intermittent dosing schedules, indicating a sustained biological activity. Conclusion: These studies provide evidence that biological activity of CHIR-258 in tumors correlates with efficacy and aids in the identification of potential biomarkers of this multitargeted receptor tyrosine kinase inhibitor. CHIR-258 exhibits properties that make it a promising candidate for clinical development in a variety of solid and hematologic malignancies.

To evaluate the pharmacokinetic (PK) properties of Bcl-2 antisense oligodeoxynucleotide G3139 when combined with the anthracycline anticancer drug doxorubicin (DOX) in a model of MDA435/LCC6 human breast cancer in severely compromised immunodeficient (SCID) mice.An orthotopic model of MDA435/LCC6 solid breast tumors was developed by bilateral implantation of passaged cells in female SCID-RAG2 mice. The G3139 plasma profile was compared for two common routes of administration (i.v. or i.p.) in single and multiple dose treatment regimens of 5 mg/kg G3139 alone or with simultaneous DOX (5 mg/kg) administration. At selected times, plasma and major organs were assayed for [3H]G3139 using scintillation counting and DOX determined using HPLC. The molecular integrity of G3139 was analyzed using SDS-PAGE. The PKs of G3139 and DOX were evaluated using a two-compartment model.G3139 administered i.v. at 5 mg/kg revealed a biexponential plasma concentration-time curve with a Cmax of 99.9 microg/ml and elimination half-lives of 0.03 h and 9.8 h, respectively, which resulted in an area under the concentration-time curve (AUC) of 15.9 microg x h/ml. G3139 administered i.p. showed a plasma absorption, distribution and elimination profile typical of this route of administration, characterized by half-lives of 0.03 h, 0.2 h and 8.9 h, respectively and a Cmax of 8.6 microg/ml. Based on AUC comparisons, the bioavailability of G3139 injected i.p. was 84% compared to i.v. administration. Subtle changes were observed in G3139 PKs after three prior i.p. doses of G3139. Specifically, a six-fold slower absorption rate, lower Cmax (6.9 microg/ml), increased Tmax (0.2 h), and an AUC of 17.4 microg x h/ml were observed, consistent with concentrations approaching saturation levels in tissue sites to which G3139 distributes. Coadministration of DOX had significant effects on the PK properties of G3139, manifested by an increased Cmax (11.2 microg/ml), higher AUC (19.7 microg x h/ml), and ninefold lower plasma clearance for single-dose G3139 administration. G3139 in plasma remained largely intact (17% degraded in plasma over 4 h), and increased plasma protein association occurred as a function of time. G3139 was detected in both healthy and tumor tissue after i.v. and i.p. administration. The highest tissue levels of G3139 were observed in the kidneys (40 microg/g), and low levels (2 microg/g) were detected in lung, heart and muscle. The rate of accumulation of G3139 in organs was dependent upon G3139 levels in plasma and the presence of coadministered DOX. Significant accumulation of G3139 was observed in solid tumors, with peak levels of approximately 5 microg G3139/g tumor, and approximately a two-to threefold tumor/muscle AUC ratio. The kinetics of G3139 accumulation in tumor tissue increased with increasing circulating G3139 concentration. The tissue distribution properties of DOX were also altered in the presence of coadministered G3139: in the presence of G3139, tumor exposure to DOX increased two-to threefold without alteration in plasma DOX PKs.These findings indicate that drug-drug interactions between G3139 and DOX are modest and favorable in that elevated tumor DOX levels are achieved without compromising G3139 tumor uptake or significantly altering plasma drug concentrations.