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Ytterbium ferrites are being used in many promising applications, such as visible-light photocatalysis, solar cells, magnetooptic devices, electro-magnetic equipment, etc., due to their fantastic ferroelectric and ferromagnetic properties. However, despite their good magnetic and radiopaque features, the use of ytterbium ferrites as multiplatform contrast agents in magnetic resonance imaging (MRI) and X-ray computed tomography (CT) is still under-developed. This is mainly due to difficulties in obtaining stable and biocompatible aqueous colloidal dispersions of ytterbium ferrite nanoparticles. In order to overcome this limitation, this work explores an eco-friendly method to directly synthesize such dispersions by liquid-assisted pulsed laser ablation of ytterbium ferrite massive targets. First, orthorhombic bulk YbFeO3 targets were obtained by a reaction-sintering method. Then, colloidal dispersions of nanoparticles were produced directly in both distilled water and ethanol by irradiating the bulk YbFeO3 targets with high-power infrared nanosecond lasers pulses. A battery of techniques has been used to characterize the as synthesized YbFeO3 targets and colloidal dispersions of YbFe nanoparticles to determine their composition, structure, magnetic properties, X-ray attenuation potentials, and colloidal properties. Moreover, the biocompatibility of the systems was also analysed by MTT cell viability assay. Results indicated that the use of distilled water as ablation medium yields colloidal dispersions consisted mainly of paramagnetic ytterbium ferrite nanoparticles. Contrarily, the use of ethanol as solvent leads to colloidal dispersions of polycrystalline nanoparticles with both ferromagnetic and paramagnetic behaviour, due to the coexistence, in each nanoparticle, of ytterbium ferrite, ytterbium oxide, and iron oxide crystalline phases. Both colloidal dispersions exhibit also high biocompatibility and suitable X-ray attenuation properties. Moreover, they show bio-safe hydrodynamic sizes (lower than 200 nm) with acceptable overall hydrodynamic polydispersity index values (under 0.4), being stable in water for several weeks. These results pave the way for the future evaluation of Yb–Fe based nanoparticles as multiplatform contrast agents in multimodal MRI and CT imaging., This research has been funded by the Spanish Ministry of Economy and Competitiveness (MINECO) and FEDERthrough the research project MAT2015-67354R, and a MSCA-IF postdoctoral fellowship from the Marie Curie action 2014 [grant 656908-NIMBLIS] of the H2020 program of the Executive Agency for Research Manages of EU Commission. Additionally, this research has been funded throught a research project (research project ULST-NANO) of the Proyectos Integradores MdM-IMEYMAT, call 2020, as well as the researcg project PID2020-118329RB-I00 funded by Spanish MCIN/AEI/10.13039/501100011033 action. Moreover, this research was also co-financed through two regional research projects funded by the Junta de Andalucía, specifically, the research project PECART-0096-2020 (Consejería Salud y Familias. JA Spain) and the research project P20_01293 (Consejería Economía, Conocimiento, Empresas y Universidad. JA Spain). In addition, Dr. Monserrat Llaguno-Munive is grateful for the funding received through the post-doctoral grant of the Consejo Nacional de Ciencia y Tecnología from Mexico (CONACYT, no 619639). In addition, we acknowledge the received technical assistance from the SC-ICYT of the University of Cádiz and the Servicio General de Apoyo a la Investigación-SAI, Universidad de Zaragoza. We also acknowledge to the Networking Research Centre on Bioengineering, Biomaterials, and Nanomedicine (CIBERBBN) (which is financed by the Instituto de Salud Carlos III (ISCIII) with assistance from the European Regional Development Fund (ERDF)) and the ICTS “NANBIOSIS”, specifically the FVPR/U20 (http://www.nanbiosis.es/portfolio/u20-in-vivo-experimentalplatform/) for providing access to the micro-CT. Finally, we want to thank the company LASING S.L. its technical support in the development of the NANO-GLAS laser system where the LA-PLA experiments were carried out.

Resumen Introduccion Un estilo de vida poco activo y una dieta inadecuada podrian influir en el desarrollo de futuras enfermedades cardiometabolicas. El objetivo principal fue determinar la asociacion entre capacidad aerobica y factores de riesgo cardiometabolicos y si dicha posible asociacion es independiente de la dieta mediterranea (DM). A su vez, se plantea como objetivo secundario estudiar el efecto combinado de la capacidad aerobica y la adherencia a la DM sobre el indice de riesgo cardiometabolico global (IRCM) en adultos. Metodos Se evaluo a 79 adultos (38% mujeres) entre 18 y 40 anos en Cadiz. Se midieron indicadores de adiposidad, presion arterial, trigliceridos, glucosa y perfil inflamatorio (interleucina-6 y factor de necrosis tumoral) y se desarrollo un IRCM. Se midio la capacidad aerobica mediante el consumo maximo de oxigeno a traves de una prueba de esfuerzo incremental en cicloergometro. La adherencia a la DM se evaluo mediante cuestionario. Se utilizo un modelo de regresion lineal para estudiar la asociacion entre capacidad aerobica y factores de riesgo cardiometabolicos con diferentes modelos de ajuste. Se analizo el efecto combinado de la capacidad aerobica y la adherencia a la DM sobre el IRCM mediante el test de ANOVA, con nivel de significacion de p Resultados Se observo asociacion negativa entre la capacidad aerobica y los factores de riesgo cardiometabolicos (todas p ≤ 0,05) en el modelo sin ajustar. La presion arterial y los trigliceridos perdieron la asociacion tras ajustar el modelo por sexo, edad y adherencia a la DM. Los participantes con alta capacidad aerobica y alta adherencia a la DM mostraron un menor IRCM (−1,083 ± 2,325 vs. 2,802 ± 1,759). Conclusiones La capacidad aerobica se asocia de forma inversa con factores de riesgo cardiometabolicos relacionados con la adiposidad, independientemente de la adherencia a la DM. Una alta adherencia a la DM podria influir en la modulacion de la presion arterial. Una alta capacidad aerobica podria reducir las consecuencias adversas de una baja adherencia a la DM.

Background In this review, we were interested to identify the wide universe of enzymes associated with epigenetic modifications, whose gene expression is regulated by miRNAs with a high relative abundance in Crohn's disease (CD) affected tissues, with the aim to determine their impact in the pathogenesis and evolution of the disease. Methods We used HMDD and Bibliometrix R-package in order to identify the miRNAs overexpressed in CD. The identified enzymes associated with epigenetic mechanisms and post-translational modifications, regulated by miRNAs upregulated in CD, were analyzed using String v11 database. Results We found 190 miRNAs with great abundance in patients with CD, of which 26 miRNAs regulate the gene expression of enzymes known to catalyze epigenetic modifications involved in essentials pathophysiological processes, such as chromatin architecture reorganization, immune response regulation including CD4+ T cells polarization, integrity of gut mucosa, gut microbiota composition and tumorigenesis. Conclusion The integrated analysis of miRNAs with a high relative abundance in patients with CD showed a combined and superimposed gene expression regulation of enzymes associated with relevant epigenetic mechanisms and that could explain, in part, the pathogenesis of CD.

An optimal design of nanoparticles suitable for biomedical applications requires proper functionalization, a key step in the synthesis of such nanoparticles, not only for subsequent crosslinking to biological targets and to avoid cytotoxicity, but also to endow these materials with colloidal stability. In this sense, a reliable characterization of the effectiveness of the functionalization process would, therefore, be crucial for subsequent bioconjugations. In this work, we have analyzed glutathione as a means to functionalize four of the most widely used nanoparticles in biomedicine, one of which is a hybrid gold-magnetic-iron-oxide nanoparticle synthetized by a simple and novel method that we propose in this article. We have analyzed the colloidal characteristics that the glutathione capping provides to the different nanoparticles and, using information on the Z-potential, we have deduced the chemical group used by glutathione to link to the nanoparticle core. We have used electron microscopy for further structural and chemical characterization of the nanoparticles. Finally, we have evaluated nanoparticle cytotoxicity, studying cell viability after incubation with different concentrations of nanoparticles, showing their suitability for biomedical applications.

Colorectal cancer (CRC) is one of the main causes of cancer death in the world. Posttranslational modifications (PTMs) have been extensively studied in malignancies due to its relevance in tumor pathogenesis and therapy. This review is focused on the dysregulation of glycosyltransferase expression in CRC and its impact in cell function and in several biological pathways associated with CRC pathogenesis, prognosis and therapeutic approaches. Glycan structures act as interface molecules between cells and their environment and in several cases facilitate molecule function. CRC tissue shows alterations in glycan structures decorating molecules, such as annexin-1, mucins, heat shock protein 90 (Hsp90), 1 integrin, carcinoembryonic antigen (CEA), epidermal growth factor receptor (EGFR), insulin-like growth factor-binding protein 3 (IGFBP3), transforming growth factor beta (TGF- ) receptors, Fas (CD95), PD-L1, decorin, sorbin and SH3 domain-containing protein 1 (SORBS1), CD147 and glycosphingolipids. All of these are described as key molecules in oncogenesis and metastasis. Therefore, glycosylation in CRC can affect cell migration, cell–cell adhesion, actin polymerization, mitosis, cell membrane repair, apoptosis, cell differentiation, stemness regulation, intestinal mucosal barrier integrity, immune system regulation, T cell polarization and gut microbiota composition; all such functions are associated with the prognosis and evolution of the disease. According to these findings, multiple strategies have been evaluated to alter oligosaccharide processing and to modify glycoconjugate structures in order to control CRC progression and prevent metastasis. Additionally, immunotherapy approaches have contemplated the use of neo-antigens, generated by altered glycosylation, as targets for tumor-specific T cells or engineered CAR (Chimeric antigen receptors) T cells.

Crohn's disease (CD) results from an aberrant immune response against the commensal microbiota in genetically susceptible hosts. However, the nature of the immune defects, the microflora involved and the genetic susceptibility remain incompletely defined and controversial. Extraintestinal manifestations occur in up to 25-35% of patients and generally precede the onset of gastrointestinal symptoms, which are often of a colonic nature and are influenced by disease activity. Renal manifestations can be considered dependent on the same immune mechanism that determines inflammatory bowel disease in CD. This review seeks to describe the current state of association between CD and kidney disease.

Background: Diabetes is a risk factor for developing Alzheimer’s disease (AD); however, the mechanism by which diabetes can promote AD pathology remains unknown. Diabetes results in diverse molecular changes in the brain, including dysregulation of glucose metabolism and loss of cerebrovascular homeostasis. Although these changes have been associated with increased Aβ pathology and increased expression of glial activation markers in APPswe/ PS1dE9 (APP/PS1) mice, there has been limited characterization, to date, of the neuroinflammatory changes associated with diabetic conditions. Results: Our analysis revealed that pathology associated with either db/db, HFD, or STZ models yielded upregulation of a broad profile of cytokines, including chemokines (e.g., MIP-1α, MIP-1β, and MCP-1) and proinflammatory cytokines, including IL-1α, IFN-γ, and IL-3. Moreover, multivariate partial least squares regression analysis showed that combined diabetic-APP/PS1 models yielded cooperatively enhanced expression of the cytokine profile associated with each diabetic model alone. Finally, in APP/PS1xdb/db mice, we found that circulating levels of Aβ1-40, Aβ1-42, glucose, and insulin all correlated with cytokine expression in the brain, suggesting a strong relationship between peripheral changes and brain pathology. Conclusions: Altogether, our multiplexed analysis of cytokines shows that Alzheimer’s and diabetic pathologies cooperate to enhance profiles of cytokines reported to be involved in both diseases. Moreover, since many of the identified cytokines promote neuronal injury, Aβ and tau pathology, and breakdown of the blood-brain barrier, our data suggest that neuroinflammation may mediate the effects of diabetes on AD pathogenesis. Therefore, strategies targeting neuroinflammatory signaling, as well as metabolic control, may provide a promising strategy for intervening in the development of diabetes-associated AD., This work was funded in part by Programa Estatal de I+D+I orientada a los Retos de la Sociedad (BFU 2016-75038-R), financed by the Agencia Estatal de Investigación (AEI) and the Fondo Europeo de Desarrollo Regional (FEDER), Ministerio de Ciencia, Innovación y Universidades, Explora Ciencia, Ministerio de Ciencia, Innovación y Universidades (BFU2017-91910-EXP), Subvención para la financiación de la investigación y la innovación biomédica y en ciencias de la salud en el marco de la Iniciativa Territorial Integrada 2014–2020 para la provincia de Cádiz, Consejeria de Salud, Junta de Andalucia, Union Europea, financed by the Fondo de Desarrollo Regional (FEDER) (PI-0008- 2017) (M.G.A.). This work was also supported by startup funds from the George W. Woodruff School of Mechanical Engineering at Georgia Tech (L.B.W.) and by the National Institutes of Health under grant number R33 ES025661 04S1. LDW was supported in part by the National Institutes of Health Cell and Tissue Engineering Biotechnology Training Grant (T32- GM008433).

Hepatitis C virus (HCV) infection is currently the most important cause of chronic viral hepatitis in the world and one of the most frequent indications for liver transplantation. HCV uses different strategies to evade the innate and adaptive immune response, and this evasion plays a key role in determining viral persistence. Several HCV viral proteins have been described as immune modulators. In this review, we will focus on the effect of HCV nucleocapsid core protein in the function of immune cells and its correlation with the findings observed in HCV chronically infected patients. Effects on immune cell function related to both extracellular and intracellular HCV core localization will be considered. This review provides an updated perspective on the mechanisms involved in HCV evasion related to one single HCV protein, which could become a key tool in the development of new antiviral strategies able to control and/or eradicate HCV infection.

Introduction Unhealthy lifestyle and inadequate diet could influence the development of future cardiometabolic disease. The main aim of this study was to determine the association between aerobic fitness and cardiometabolic risk factors in adults, whether this relation is depends of adherence to Mediterranean diet (MD). A secondary aim was to study the combined effect of aerobic capacity and adherence to MD on global cardiometabolic risk score (CMRS). Method A total of 79 adults (38% women) enrolled between 18–40 year from Cadiz. We measured adiposity indicators, blood pressure, triglycerides, glucose and inflammatory profile (interleukin-6 and tumor necrosis factor) and was computed (CMRS). Aerobic fitness was measured by maximal oxygen comsuption through an incremental stress test by cycleergometer. The MD patterns was measured using the questionnaire of adherence to MD. The association between aerobic fitness and cardiometabolic risk factors was examined using a lineal regression and it was adjusted for different confounders. CMRS on the lifestyle was analyzed using the ANOVA test, with statistical significance level of p Results Linear regression showed inverse association between aerobic fitness and cardiometabolic risk factors (all p ≤ 0.05) in the model without adjustment. Blood pressure and triglycerides lost the association after adjust model for sex, age, and adherence to MD. Participants with high aerobic fitness and high adherence to MD show a lowest CMRS (−1.083 ± 2.325 vs. 2.802 ± 1.759). Conclusions Aerobic fitness was inversely associated with fatness risk factors, that relationship is independent to adherence to MD. A high adherence to MD could modulate blood pressure. A combination of high aerobic capacity and high adherence to MD could reduce the adverse consecuence of a low adherencie to MD.

The aim of this study was to identity in silico the relationships among microRNAs (miRNAs) and genes encoding transcription factors, ubiquitylation, DNA methylation, and histone modifications in systemic lupus erythematosus (SLE). To identify miRNA dysregulation in SLE, we used miR2Disease and PhenomiR for information about miRNAs exhibiting differential regulation in disease and other biological processes, and HMDD for information about experimentally supported human miRNA–disease association data from genetics, epigenetics, circulating miRNAs, and miRNA–target interactions. This information was incorporated into the miRNA analysis. High-throughput sequencing revealed circulating miRNAs associated with kidney damage in patients with SLE. As the main finding of our in silico analysis of miRNAs differentially expressed in SLE and their interactions with disease-susceptibility genes, post-translational modifications, and transcription factors; we highlight 226 miRNAs associated with genes and processes. Moreover, we highlight that alterations of miRNAs such as hsa-miR-30a-5p, hsa-miR-16-5p, hsa-miR-142-5p, and hsa-miR-324-3p are most commonly associated with post-translational modifications. In addition, altered miRNAs that are most frequently associated with susceptibility-related genes are hsa-miR-16-5p, hsamiR- 374a-5p, hsa-miR-34a-5p, hsa-miR-31-5p, and hsa-miR-1-3p.

Hepatitis C virus core protein is a highly basic viral protein that multimerizes with itself to form the viral capsid. When expressed in CD4(+) T lymphocytes, it can induce modifications in several essential cellular and biological networks. To shed light on the mechanisms underlying the alterations caused by the viral protein, we have analyzed HCV-core subcellular localization and its associations with host proteins in Jurkat T cells. In order to investigate the intracellular localization of Hepatitis C virus core protein, we have used a lentiviral system to transduce Jurkat T cells and subsequently localize the protein using immunoelectron microscopy techniques. We found that in Jurkat T cells, Hepatitis C virus core protein mostly localizes in the nucleus and specifically in the nucleolus. In addition, we performed pull-down assays combined with Mass Spectrometry Analysis, to identify proteins that associate with Hepatitis C virus core in Jurkat T cells. We found proteins such as NOLC1, PP1 gamma, ILF3, and C1QBP implicated in localization and/or traffic to the nucleolus. HCV-core associated proteins are implicated in RNA processing and RNA virus infection as well as in functions previously shown to be altered in Hepatitis C virus core expressing CD4(+) T cells, such as cell cycle delay, decreased proliferation, and induction of a regulatory phenotype. Thus, in the current work, we show the ultrastructural localization of Hepatitis C virus core and the first profile of HCV core associated proteins in T cells, and we discuss the functions and interconnections of these proteins in molecular networks where relevant biological modifications have been described upon the expression of Hepatitis C virus core protein. Thereby, the current work constitutes a necessary step toward understanding the mechanisms underlying HCV core mediated alterations that had been described in relevant biological processes in CD4(+) T cells.

Non-T cell activation linker is an adaptor protein that is tyrosine phosphorylated upon cross-linking of immune receptors expressed on B lymphocytes, NK cells, macrophages, basophils, or mast cells, allowing the recruitment of cytosolic mediators for downstream signaling pathways. Fas receptor acts mainly as a death receptor, and when cross-linked with Fas ligand, many proteins are proteolytically cleaved, including several signaling molecules in T and B cells. Fas receptor triggering also interferes with TCR intracellular signals, probably by means of proteolytic cleavage of several adaptor proteins. We have previously found that the adaptor linker for activation of T cells, evolutionarily related to non-T cell activation linker, is cleaved upon proapoptotic stimuli in T lymphocytes and thymocytes, in a tyrosine phosphorylation-dependent fashion. Here, we describe non-T cell activation linker proteolytic cleavage triggered in human B cells and monocytes by Fas cross-linking and staurosporine treatment. Non-T cell activation linker is cleaved, producing an N-terminal fragment of ∼22 kDa, and such cleavage is abrogated in the presence of caspase 8/granzyme B and caspase 3 inhibitors. Moreover, we have identified an aspartic acid residue at which non-T cell activation linker is cleaved, which similar to linker for activation of T cells, this aspartic acid residue is located close to tyrosine and serine residues, suggesting an interdependence of phosphorylation and proteolytic cleavage. Consistently, induction of non-T cell activation linker phosphorylation by pervanadate inhibits its cleavage. Interestingly, the truncated isoform of non-T cell activation linker, generated after cleavage, has a decreased signaling ability when compared with the full-length molecule. Altogether, our results suggest that cleavage of transmembrane adaptors constitutes a general mechanism for signal termination of immune receptors.

Adaptive T cell responses are critical for controlling HCV infection. While there is clinical evidence of a relevant role for regulatory T cells in chronic HCV-infected patients, based on their increased number and function; mechanisms underlying such a phenomena are still poorly understood. Accumulating evidence suggests that proteins from Hepatitis C virus can suppress host immune responses. We and others have shown that HCV is present in CD4+ lymphocytes from chronically infected patients and that HCV-core protein induces a state of unresponsiveness in the CD4+ tumor cell line Jurkat. Here we show that CD4+ primary T cells lentivirally transduced with HCV-core, not only acquire an anergic phenotype but also inhibit IL-2 production and proliferation of bystander CD4+ or CD8+ T cells in response to anti-CD3 plus anti-CD28 stimulation. Core-transduced CD4+ T cells show a phenotype characterized by an increased basal secretion of the regulatory cytokine IL-10, a decreased IFN-γ production upon stimulation, as well as expression of regulatory T cell markers, CTLA-4, and Foxp3. A significant induction of CD4+CD25+CD127(low)PD-1(high)TIM-3(high) regulatory T cells with an exhausted phenotype was also observed. Moreover, CCR7 expression decreased in HCV-core expressing CD4+ T cells explaining their sequestration in inflamed tissues such as the infected liver. This work provides a new perspective on de novo generation of regulatory CD4+ T cells in the periphery, induced by the expression of a single viral protein.

HCV (hepatitis C virus) infection is a serious health care problem that affects more than 170 million people worldwide. Viral clearance depends on the development of a successful cellular immune response against the virus. Interestingly, such a response is altered in chronically infected patients, leading to chronic hepatitis that can result in liver fibrosis, cirrhosis and hepatocellular carcinoma. Among the mechanisms that have been described as being responsible for the immune suppression caused by the virus, Treg-cells (regulatory T-cells) are emerging as an essential component. In the present work we aim to study the effect of HCV-core protein in the development of T-cells with regulatory-like function. Using a third-generation lentiviral system to express HCV-core in CD4+ Jurkat T-cells, we describe that HCV-core-expressing Jurkat cells show an up-regulation of FOXP3 (forkhead box P3) and CTLA-4 (cytotoxic T-lymphocyte antigen-4). Moreover, we show that HCV-core-transduced Jurkat cells are able to suppress CD4+ and CD8+ T-cell responses to anti-CD3 plus anti-CD28 stimulation.

El síndrome de Nager (OMIM 154400) es una enfermedad genética que incluye principalmente hipoplasia de los radios y signos faciales como hipoplasia malar, fisuras palpebrales antimongoloides, micrognatia y defectos en pabellones auriculares. En este trabajo se reporta una paciente que consulta al servicio de dismorfología por presentar falta de desarrollo de dedos pulgares y disminución de la audición. Durante la evaluación genética se encuentra una paciente femenina de catorce años de edad, hija de padres sanos con antecedentes de consanguinidad y endogamia. Embarazo de curso normal sin exposición a teratógenos y parto normal. El examen clínico mostró facies atípica por presencia de fisuras palpebrales antimongoloides, puente nasal alto, orejas pequeñas de implantación baja y micrognatia. En miembros superiores presentó aplasia de pulgares y una marcada limitación en los movimientos de la articulación del codo. Durante el análisis familiar se encuentra que su hermana presenta signos faciales similares a la paciente y una hipoplasia moderada de los dedos pulgares en los miembros superiores. Estudios radiológicos mostraron la presencia de radio y sinostosis radiocubital proximal. Además, el estudio citogenético reportó cariotipo normal y el análisis de audiometría reporta una disminución leve de la agudeza auditiva. Con los hallazgos clínicos y radiológicos, la paciente y su hermana se clasifican como un síndrome de Nager o disostosis acrofacial. Esta patología cursa con heterogeneidad genética por su presentación esporádica en un alto porcentaje de pacientes. En algunas familias se asocia a una proteína (ZNF-37) en el locus 9q32 del cromosoma 9 y se hereda de forma autosómica dominante con penetrancia variable y marcada expresividad; además, se han descrito casos de hermanos afectados con padres normales, lo que se asocia a una herencia autosómica recesiva. También se ha asociado con una deleción parcial en el brazo largo del cromosoma 1 y con una brecha de cromatina en la banda 3p14. En esta familia, por el antecedente de consanguinidad y padres sin signos que demuestren alguna expresión de la enfermedad, se clasifica como una herencia autosómica recesiva y se hace énfasis en la importancia del diagnóstico diferencial con el síndrome de Treacher-Collins para efectos del manejo y consejería genética.

Advances in the life sciences are now closely linked to the availability of new experimental tools that allow for the manipulation of biomolecules and the study of biological processes at the molecular level. In this context, we have optimized a synthesis process to obtain glutathione-capped fluorescent CdS nanoparticles to specifically detect Glutathione S-Transferase (GST) -tagged proteins. Using our method, based on five different heating steps, brightly fluorescent and biocompatible CdS quantum dots of different sizes can be obtained. QD optical behaviour has been evaluated studying both absorbance and fluorescence. For all the samples, the excitonic absorption onset clearly shows a blue shift at 512nm in comparison with that of bulk CdS, due to the quantum confinement effect. At increased average sizes of the nanocrystal, the emission fluorescent band shows a red shift, from 440nm to 540nm. Among different QD solutions, we demonstrate an expansion of the emission range up to ~100 nm, thus improving their features as biomarkers. Moreover we show that optimized glutathione-capped quantum dots can directly bind GST blotted onto polyvinylidene difluoride (PVDF) membranes, and thus are suitable for the direct detection of GST fusion proteins.