Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports

In the present research effort, production of derivatives of cardosin A (a plant protease) encompassing full stabilization of its dimeric structure has been achieved, via covalent, multi-subunit immobilization onto highly activated agarose-glutaraldehyde supports. Boiling such enzyme derivatives in the presence of sodium dodecyl sulfate and β-mercaptoethanol did not lead to leaching of enzyme, thus providing evidence for the effectiveness of the attachment procedure. Furthermore, the cardosin A derivatives prepared under optimal conditions presented ca. half the specific activity of the enzyme in soluble form, and were successfully employed at laboratory-scale trials to perform (selective) hydrolysis of α-lactalbumin (α-La), one of the major proteins in bovine whey. Hydrolysates of α-La were assayed for by the OPA method, as well as by FPLC, SDS–PAGE and HPLC. Thermal inactivation of the immobilized cardosin A was also assessed at 40, 50 and 55 °C; at these temperatures, no thermal denaturation took place during incubation for 48 h. The highest degree of hydrolysis was attained by 5 h reaction, at 55 °C and pH 5.2. SDS–PAGE of α-La hydrolysates displayed bands corresponding to low molecular weight peptides. Our results suggest that cardosin A in immobilized form is a good candidate to bring about proteolysis in the dairy industry, namely in whey processing

Avaliação da Cinética de Degradação Térmica de Membranas de Acetato de Celulose Híbridas usando Métodos Isoconversionais

Cellulose acetate membranes are widely used in industry, emphasizing water purification processes, such as desalination. With some limiting mechanical properties, the synthesis of hybrid membranes appears as an alternative for developing high-performance materials. For its application, knowledge of thermal stability is crucial. In this work, the thermal degradation kinetics of AC-SiO2-(CH2)3NH2 hybrid cellulose acetate membranes are evaluated from thermogravimetric analysis, at three heating rates, 5, 10, and 20°C/min. The isoconversional methods proposed by Kissinger, Flynn-Wall-Ozawa, and Friedman were used for the present study of degradation kinetics. It was observed that insertion of silicon to polymeric structure promoted thermal stability to the membrane, presenting higher activation energy than pure cellulose acetate membrane, increasing from 240.28 to 1039.01 KJ/mol, using the method of Friedman. In contrast, the increase in nitrogen concentration decreases its thermal stability compared to the cellulose acetate membrane with incorporated silicon, reducing the activation energy from 1039.01 to 250.50 KJ/mol. However, it is more stable than the pure cellulose acetate membrane. The evaluation carried out in this study explained the influence of the minimum variation in the chemical composition against the thermal stability of hybrid membranes, being a factor of great importance for its application.As membranas de acetato de celulose são amplamente utilizadas na indústria, nos processos de purificação de água, principalmente na dessalinização. Com algumas propriedades limitantes, a síntese de membranas híbridas aparece como uma alternativa para o desenvolvimento de materiais de alto desempenho. Para sua aplicação, o conhecimento da estabilidade térmica é crucial. Neste trabalho, a cinética de degradação térmica de membranas híbridas de acetato de celulose AC-SiO2-(CH2)3NH2 é avaliada a partir de análise termogravimétrica, em três taxas de aquecimento, 5; 10 e 20 °C/min. Os métodos isoconversionais propostos por Kissinger, Flynn-Wall-Ozawa e Friedman foram usados para o presente estudo da cinética de degradação. Observou-se que a inserção do silício na estrutura polimérica promoveu estabilidade térmica à membrana, apresentando energia de ativação superior à membrana de acetato de celulose puro, passando de 240,28 para 1039,01 kJ/mol, pelo método de Friedman. Em contraste, o aumento na concentração de nitrogênio diminui sua estabilidade térmica em relação à membrana de acetato de celulose com silício incorporado, reduzindo a energia de ativação de 1039,01 para 250,50 kJ/mol. No entanto, é mais estável do que a membrana de acetato de celulose pura. A avaliação realizada neste estudo explicou a influência da variação mínima na composição química contra a estabilidade térmica de membranas híbridas, sendo um fator de grande importância para sua aplicação

Extraction, purification and characterization of galactomannans from non-traditional sources

This work presents a methodology for the extraction of galactomannans from seeds of four different species of Leguminosae (Adenanthera pavonina, Caesalpinia pulcherrima, Gleditsia triacanthos and Sophora japonica) to be used e.g. in the food and biomedical industries. The galactomannans were obtained by aqueous extraction followed by a precipitation with ethanol. This methodology is simpler and easier to perform than other existing extraction and purification methodologies, and because it avoids the use of organic solvents (other than ethanol), it is able to generate food grade substances and is environmentally friendlier. The yield of extraction in different stages of the process, monosaccharide composition, as well as physical and chemical parameters of the isolated galactomannans were determined and compared with previously published results. The mannose/galactose ratio of the extracted galactomannans ranged from 1.35 (A. pavonina) to 5.75 (S. japonica). The intrinsic viscosity ranged from 11.34 dL/g (C. pulcherrima) to 8.74 dL/g (S. japonica), while the viscosity average molecular mass ranged between 1.81 * 10 6Da and 1.17 * 10 6Da (A. pavonina > C. pulcherrima > G. triacanthos > S. japonica). The results confirm the suitability of the extraction and purification procedure to obtain galactomannans from non-traditional sources.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brazil)Fundação para a Ciência e Tecnologia (FCT)ALFA VALNATURA Project of Europe Aid Cooperation Office (EU

Atividades terapêuticas em contexto de saúde mental e psiquiatria: Uma reflexão sobre a ação

No âmbito do Estágio V - Enfermagem de Saúde Mental e Psiquiatria, no processo de cuidados à pessoa com doença mental, em fase aguda, em contexto de internamento, foi possível experienciar o desenvolvimento de Atividades terapêuticas. Segundo Zimerman e Osório (1997, p.35), as atividades terapêuticas tem como objetivo “proporcionar ao indivíduo o máximo desempenho e autonomia nas suas funções pessoais, sociais e profissionais”. Após a realização da ação, a “reflexão sobre ela só tem sentido para compreender, aprender e integrar o que acontece” (Perrenoud, 2011, p.31). O processo reflexivo facilita a compreensão da relação existente, entre os conceitos teóricos e situações clínicas experienciadas (Potter e Perry, 2003). Objetivo Partilhar a reflexão sobre o processo de planeamento, implementação e avaliação de atividades terapêuticas em contexto de saúde mental e psiquiatria. Materiais e métodos Relativamente à metodologia optou-se pela reflexão sobre a ação (Santos & Fernandes, 2004). Nessa perspetiva, as etapas orientadoras para uma reflexão estruturada são: Descrição da ação tal e qual é percebida; Reflexão; Síntese/Avaliação da reflexão utilizada (Santos & Fernandes, 2004, p.61). Resultados A apresentação dos resultados segue as etapas previstas na metodologia. As atividades terapêuticas desenvolvidas foram: “Expressão pessoal” e “Recreação/Lazer” (Melo-Dias, 2014). No planeamento adotaram-se critérios específicos de inclusão e exclusão bem como a intenção de livre participação. A implementação das atividades respeitou a Norma nº011/2020 (DGS, 2020). No términus das atividades foram auscultadas as opiniões de cada participante sobre os contributos das mesmas. Conclusões As atividades terapêuticas desempenham um forte contributo para a reinserção social da pessoa com doença mental. A implementação destas atividades integradas na prestação de cuidados global à pessoa com doença mental traduziu-se na integração de aprendizagens necessárias ao desenvolvimento pessoal e profissional. Referências Bibliográficas Direção Geral da Saúde. (2020). Covid-19: FASE DE MITIGAÇÃO Saúde Mental. Norma nº 011/2020 de 18/04/2020; Lisboa:Direção Geral da Saúde. Madeira, A. C. S. C. M. (2014). O estudante de enfermagem no processo de cuidados em contexto de ensino clínico (Doctoral dissertation, Tese de Doutoramento, Universidade do Porto, Porto). Disponível em: https://repositorioaberto. up. pt/bitstream/10216/78973/2/35058. pdf). Melo-Dias, C., Rosa, A., & Pinto, A. (2014). ATIVIDADES DE OCUPAÇÃO TERAPÊUTICA--INTERVENÇÕES DE ENFERMAGEM ESTRUTURADAS EM REABILITAÇÃO PSICOSSOCIAL. Portuguese Journal of Mental Health Nursing/Revista Portuguesa de Enfermagem de Saude Mental, (11). Perrenoud, P. (2011). A Prática Reflexiva do Professor: Profissionalização e Razão Pedagógica. São Paulo: Artmed Editores, SA. Potter e Perry (2003). Fundamentos de Enfermagem: Conceitos e Procedimentos. Santos,E.;Fernandes,A.(2004)–Prática Reflexiva: guia para a reflexão estruturada. In: Revista de Enfermagem Referência Coimbra – ISSN0874-0283-Nº11(Mar.2004), p.59-62. Disponível em: https://rr.esenfc.pt/rr/index.php?module=rr&target=publicationDetails&pesquisa=&id_artigo=2064&id_revista=5&id_edicao= 10 Zimerman, D. O., & Osorio, E. (1997). LC Como trabalhamos com grupos. Porto Alegre.N/

One-pot intercalation strategy for the encapsulation of a CO-releasing organometallic molecule in a layered double hydroxide

The photoactivatable CO‐releasing molecule (photoCORM) [Mo(CO)3(CNCH2COOH)3] (ALF795) has been incorporated into a Zn,Al layered double hydroxide (LDH) host by a coprecipitation synthesis strategy. Powder X‐ray diffraction (PXRD) of the resultant material Zn,Al‐ALF795 showed that the ALF795 guest molecules assembled into a monolayer to give a basal spacing of 16.0 Å. FTIR and 13C{1H} CP MAS NMR spectroscopy confirmed that the molecular structure of the tricarbonyl complex was retained upon intercalation. Scanning electron microscopy (SEM), energy‐dispersive X‐ray spectroscopy (EDS) and elemental analyses confirmed the phase‐purity of Zn,Al‐ALF795. The myoglobin assay was used to demonstrate that intercalated ALF795 retains the photoactive behavior of the free CORM, with a substantial fraction (42 %) of the high CO payload (2.46 mmol g–1) being released after exposure to UV light for 3 h under simulated physiological conditions. In addition, gas chromatography was used to track sequential light‐ and H2O2‐triggered decarbonylation of free and intercalated ALF795. In biological buffer solution (HEPES), less than 2 % Mo leaching from Zn,Al‐ALF795 took place after 5 h, showing the strong capacity of the LDH host to retain the unaltered complex and decarbonylation fragments.publishe

Preclinical pharmacokinetics and biodistribution of anticancer dinuclear Palladium(II)-Spermine Complex (Pd2Spm) in mice

Palladium-based compounds are regarded as potential analogs to platinum anticancer drugs with improved properties. The present study assessed the pharmacokinetics and biodistribution of a dinuclear palladium(II)-spermine chelate (Pd2Spm), which has previously been shown to possess promising in vitro activity against several therapy-resistant cancers. Using inductively coupled plasma-mass spectrometry, the kinetic profiles of palladium/platinum in serum, serum ultrafiltrate and tissues (kidney, liver, brain, heart, lungs, ovaries, adipose tissue and mammary glands) were studied in healthy female Balb/c mice after a single intraperitoneal bolus injection of Pd2Spm (3 mg/kg bw) or cisplatin (3.5 mg/kg bw) between 0.5 and 48 h post-injection. Palladium in serum exhibited biphasic kinetics with a terminal half-life of 20.7 h, while the free palladium in serum ultrafiltrate showed a higher terminal half-life than platinum (35.5 versus 31.5 h). Palladium was distributed throughout most of the tissues except for the brain, with the highest values in the kidney, followed by the liver, lungs, ovaries, adipose tissue and mammary glands. The in vitro cellular accumulation was also evaluated in breast cancer cells, evidencing a passive diffusion as a mechanism of Pd2Spm’s cellular entry. This study reports, for the first time, the favorable pharmacokinetics and biodistribution of Pd2Spm, which may become a promising pharmacological agent for cancer treatmentinfo:eu-repo/semantics/publishedVersio

WNT6 is a novel oncogenic prognostic biomarker in human glioblastoma

Glioblastoma (GBM) is a universally fatal brain cancer, for which novel therapies targeting specific underlying oncogenic events are urgently needed. While the WNT pathway has been shown to be frequently activated in GBM, constituting a potential therapeutic target, the relevance of WNT6, an activator of this pathway, remains unknown. Methods: WNT6 protein and mRNA levels were evaluated in GBM. WNT6 levels were silenced or overexpressed in GBM cells to assess functional effects in vitro and in vivo. Phospho-kinase arrays and TCF/LEF reporter assays were used to identify WNT6-signaling pathways, and significant associations with stem cell features and cancer-related pathways were validated in patients. Survival analyses were performed with Cox regression and Log-rank tests. Meta-analyses were used to calculate the estimated pooled effect. Results: We show that WNT6 is significantly overexpressed in GBMs, as compared to lower-grade gliomas and normal brain, at mRNA and protein levels. Functionally, WNT6 increases typical oncogenic activities in GBM cells, including viability, proliferation, glioma stem cell capacity, invasion, migration, and resistance to temozolomide chemotherapy. Concordantly, in in vivo orthotopic GBM mice models, using both overexpressing and silencing models, WNT6 expression was associated with shorter overall survival, and increased features of tumor aggressiveness. Mechanistically, WNT6 contributes to activate typical oncogenic pathways, including Src and STAT, which intertwined with the WNT pathway may be critical effectors of WNT6-associated aggressiveness in GBM. Clinically, we establish WNT6 as an independent prognostic biomarker of shorter survival in GBM patients from several independent cohorts. Conclusion: Our findings establish WNT6 as a novel oncogene in GBM, opening opportunities to develop more rational therapies to treat this highly aggressive tumor.FCT - Foundation for Science and Technology (PTDC/SAU-GMG/113795/2009 and IF/00601/2012 to B.M.C.; SFRH/BD/92786/2013 to C.S.G.; SFRH/BD/88121/2012 to J.V.C.; SFRH/BD/81042/2011 to M.P.; SFRH/BD/93443/2013 to S.Q.) and Fundação Calouste Gulbenkian (B.M.C.), by FEDER funds through the Operational Programme Competitiveness Factors - COMPETE and National Funds through FCT under the project POCI-01-0145-FEDER-007038; by the project NORTE-01-0145-FEDER-000013 and NORTE-01-0246-FEDER-000012, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF); and by the project NORTE-01-0145-FEDER-000023, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER)info:eu-repo/semantics/publishedVersio

CD44v6 high membranous expression is a predictive marker of therapy response in gastric cancer patients

© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).In gastric cancer (GC), biomarkers that define prognosis and predict treatment response remain scarce. We hypothesized that the extent of CD44v6 membranous tumor expression could predict prognosis and therapy response in GC patients. Two GC surgical cohorts, from Portugal and South Korea (n = 964), were characterized for the extension of CD44v6 membranous immuno-expression, clinicopathological features, patient survival, and therapy response. The value of CD44v6 expression in predicting response to treatment and its impact on prognosis was determined. High CD44v6 expression was associated with invasive features (perineural invasion and depth of invasion) in both cohorts and with worse survival in the Portuguese GC cohort (HR 1.461; 95% confidence interval 1.002-2.131). Patients with high CD44v6 tumor expression benefited from conventional chemotherapy in addition to surgery (p < 0.05), particularly those with heterogeneous CD44v6-positive and -negative populations (CD44v6_3+) (p < 0.007 and p < 0.009). Our study is the first to identify CD44v6 high membranous expression as a potential predictive marker of response to conventional treatment, but it does not clarify CD44v6 prognostic value in GC. Importantly, our data support selection of GC patients with high CD44v6-expressing tumors for conventional chemotherapy in addition to surgery. These findings will allow better stratification of GC patients for treatment, potentially improving their overall survival.This work was funded by FEDER-Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020–Operacional Programme for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through FCT–Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Inovação in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274). This work was also financed by the projects NORTE-01-0145-FEDER-000003 and NORTE-01-0145-FEDER-000029, supported by the Norte Portugal Regional Programme (NORTE 2020) under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF); project POCI-01-0145-FEDER-016390 and SAICTPAC/0022/2015, funded by ERDF, POCI, and FCT; project PTDC/CTM-NAN/120958/2010, from FCT; and by project PTDC/BTM-TEC/30164/2017 funded by ERDF funds through the COMPETE 2020–POCI, Portugal 2020, and by FCT. Salary support to G.M.A. by PTDC/BTM-TEC/30164/2017 project; C.P. was supported by the grant SFRH/BD/113031/2015 from FCT.info:eu-repo/semantics/publishedVersio