99mTc(I)-Labeled His-Tagged Proteins: Impact in the Development of Novel Imaging Probes and in Drug Discovery.

Technetium-99m (99mTc) remains the cornerstone of nuclear medicine for single photon emission computed tomography (SPECT) due to its widespread availability and chemical and physical features. Its multiple oxidation states allow for the design and production of radiopharmaceuticals with versatile properties, namely in terms of pharmacokinetic profile. 99mTc(V) is the most common oxidation state, but 99mTc(I) gained traction after the pioneering work of Alberto and colleagues, which resulted in the introduction of the organometallic core fac-[99mTc(CO)3(H2O)3]+. This core is readily available from [99mTcO4]- and displays three labile water molecules that can be easily swapped for ligands with different denticity and/or donor atoms in aqueous environment. This makes it possible to radiolabel small molecules as well as high molecular weight molecules, such as antibodies or other proteins, while assuring biological activity. Direct radiolabelling of those proteins with fac-[99mTc(CO)3]+ under mild conditions is accomplished through incorporation of a polyhistidine tag (His-tag), a commonly used tag for purification of recombinant proteins. This review aims to address the direct radiolabelling of His-tagged macromolecules with fac-[99mTc(CO)3]+ for development of molecular imaging agents and the impact of this technology in the discovery and development of imaging and/or therapeutic agents towards clinical application.

Pharmacovigilance teaching and learning: a mixed cross-sectional analysis of the Portuguese public higher education system.

Pharmacovigilance stands out for its importance in obtaining existing knowledge about medicine and patient safety and should be recognized as a continuous line of study. It constitutes a highly relevant component in the activities of health professionals, with spontaneous notification of suspected adverse drug reactions being its main emphasis. The underreporting that persists can be overcome through continuous professional development programs, reinforcing theoretical and practical knowledge in the curricular plans of health courses. As a result, more educated professionals will also allow citizens to recognize the importance of pharmacovigilance. The main objective of this study was to describe and characterize the teaching-learning process of pharmacovigilance in Portugal, analyzing the knowledge, perceptions and attitudes of students and health professionals. In total, ninety-three curricular unit forms of the seventeen healthcare courses included were analyzed, among which only three referred to pharmacovigilance as mandatory and thirty-nine did not address any keywords. The questionnaire applied was answered by 650 participants, both students (62%) and professionals (38%). Approximately 84.4% of the students and 54.7% of the professionals affirmed that they had never spontaneously reported an adverse drug reaction. Only 24.6% of the students and 17.8% of professionals referred to the existence of specific course content dedicated to pharmacovigilance in their coursework. In view of these results, it is evident that there is a need for a wider reflection regarding the further training and constant update of practicing professionals as well as in diverse health institutions, investing in the creation of an academic curriculum that integrates pharmacovigilance in healthcare courses.

Tailored Viral-like Particles as Drivers of Medical Breakthroughs.

Despite the recognized potential of nanoparticles, only a few formulations have progressed to clinical trials, and an even smaller number have been approved by the regulatory authorities and marketed. Virus-like particles (VLPs) have emerged as promising alternatives to conventional nanoparticles due to their safety, biocompatibility, immunogenicity, structural stability, scalability, and versatility. Furthermore, VLPs can be surface-functionalized with small molecules to improve circulation half-life and target specificity. Through the functionalization and coating of VLPs, it is possible to optimize the response properties to a given stimulus, such as heat, pH, an alternating magnetic field, or even enzymes. Surface functionalization can also modulate other properties, such as biocompatibility, stability, and specificity, deeming VLPs as potential vaccine candidates or delivery systems. This review aims to address the different types of surface functionalization of VLPs, highlighting the more recent cutting-edge technologies that have been explored for the design of tailored VLPs, their importance, and their consequent applicability in the medical field.

Mobile apps for quick adverse drug reaction report: A scoping review.

PURPOSE: Spontaneous notification systems are essential in a post-marketing safety context. However, using this method, only about 6% of all adverse drug reactions are notified. To overcome this sub-notification problem, new methods need to be developed to improve and facilitate reporting. In this sense, the use of digital media, mainly medical mobile apps, has been presented as a powerful tool, including in pharmacovigilance. We performed a scope review to identify the available apps used to report adverse drug reactions around the world to eventually identify which of them best fits the Portuguese pharmacovigilance system. METHODS: The Joanna Briggs Institute guidelines were considered, and the framework proposed by Arksey and O'Malley was followed. All the articles that met the inclusion criteria were examined for this review. When the studies lacked in information about the app, Google was used to enhance the search for further information. RESULTS: A final number of five articles were included, revealing seven implemented mobile apps for adverse drug reaction report (Medwatcher, VigiBIP, Yellow Card, Bijwerking, Halmed, Med Safety, and ADR PvPi). These apps are implemented in the United States, France, United Kingdom, The Netherlands, Croatia, and India. Med Safety was originally designed for multi-region use and is implemented in 12 low and middle-income countries. CONCLUSIONS: Apps are easier and faster ways of reporting. The integration of such a tool in an individual care plan would allow to maintain a complete electronic health record at both individual and global level and could be eventually seen as an added value by both health professionals and patients. A country specific version of the WEB-RADR could be a solution for Portugal, in order to introduce an app to notify ADRs at the national level, due previous successful experiences in European countries.

A CASE OF KRETSCHMER’S SENSITIVE DELUSION RELATED TO COVID-19 PANDEMIC.

The COVID-19 pandemic has already infected more than 182 million people and killed more than 4 million all over the globe. In addition to its direct health effects, lockdowns and other draconian public health measures, along with an expected economic crisis of unprecedent magnitude, unpre- dictable social effects are being generated.

Consumption of antidiabetic medicines in Portugal: results of a temporal data analysis of a thirteen-year study (2005-2017).

BACKGROUND: Studies of drug utilization in patients with diabetes, a chronic disease that can be treated with a wide range of available medicines, have attracted substantial social and clinical interest. OBJECTIVE: To characterize antidiabetic medicine consumption between 2005 and 2017, to evaluate the trends of these medicines in mainland Portugal, and to compare district consumption. An additional objective was to perform a statistical analysis on drug consumption in different regions of Portugal. METHODS: A descriptive, longitudinal observational study; the setting was mainland Portugal ( excluding Azores and Madeira). Each medicine has a respective defined daily dose (DDD). The sum of the DDD, provides the annual consumption in terms of the DDD for each district each year. When calculating the annual average for the resident district population and the number of days in a year, the denominator is expressed as 1000 inhabitants per day (TID). MAIN OUTCOME MEASURE: The DDD/TID for mainland Portugal (for all districts) between 2005 and 2017 for antidiabetic medicines. Information was obtained from the official database of prescription medicine invoices with reimbursement in mainland Portugal. RESULTS: In mainland Portugal, the antidiabetic medicine consumption was 49.3 DDD/TID in 2005 and 88.2 DDD/TID in 2017. The consumption of insulins and their analogs increased from 10.8% to 17.4% compared to the total consumption of antidiabetic medicines. In 2017, the level of biguanide consumption was 23.1 DDD/TID, that of sulphonylurea consumption was 15.8 DDD/TID, that of DPP-4 inhibitor consumption was 6.8 DDD/TID, and that of SGLT2 inhibitor consumption was 3.0 DDD/TID. The oral consumption of fixed-dose combinations reached 21.4 DDD/TID. After employing a geographical division between north and south and between coastal and inland regions, the consumption of several different drugs showed statistically significant differences. CONCLUSIONS: When comparing 2017 with 2005, the panorama was quite different, with higher levels of consumption of antidiabetic medicines, insulins and their analogs, noninsulin medicines, long-acting and fast-acting insulins and their analogs, metformin, DPP-4 inhibitors and, mainly, metformin combined with a DPP-4 inhibitor. The SGLT2 inhibitors achieved a representative consumption. Different consumption patterns may be related to sociodemographic factors or to clinical practices.

Multiple Bacteria Identification in the Point-of-Care: an Old Method Serving a New Approach.

The accurate diagnosis of bacterial infections is of critical importance for effective treatment decisions. Due to the multietiologic nature of most infectious diseases, multiplex assays are essential for diagnostics. However, multiplexability in nucleic acid amplification-based methods commonly resorts to multiple primers and/or multiple reaction chambers, which increases analysis cost and complexity. Herein, a polymerase chain reaction (PCR) offer method based on a universal pair of primers and an array of specific oligonucleotide probes was developed through the analysis of the bacterial 16S ribosomal RNA gene. The detection system consisted of DNA hybridization over an array of magnetoresistive sensors in a microfabricated biochip coupled to an electronic reader. Immobilized probes interrogated single-stranded biotinylated amplicons and were obtained using asymmetric PCR. Moreover, they were magnetically labelled with streptavidin-coated superparamagnetic nanoparticles. The benchmarking of the system was demonstrated to detect five major bovine mastitis-causing pathogens: Escherichia coli, Klebsiella sp., Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae. All selected probes proved to specifically detect their respective amplicon without significant cross reactivity. A calibration curve was performed for S. agalactiae, which demonstrates demonstrating a limit of detection below 30 fg/µL. Thus, a sensitive and specific multiplex detection assay was established, demonstrating its potential as a bioanalytical device for point-of-care applications.

Go with the flow: advances and trends in magnetic flow cytometry.

The growing need for biological information at the single cell level has driven the development of improved cytometry technologies. Flow cytometry is a particularly powerful method that has evolved over the past few decades. Flow cytometers have become essential instruments in biomedical research and routine clinical tests for disease diagnosis, prognosis, and treatment monitoring. However, the increasing number of cellular parameters unveiled by genomic, proteomic, and metabolomic data platforms demands an augmented multiplexability. Also, the need for identification and quantification of relevant biomarkers at low levels requires outstanding analytical sensitivity and reliability. In addition, growing awareness of the advantages associated with miniaturization of analytical devices is pushing forward the progress in integrated and compact, microfluidic-based devices at the point-of-care. In this context, novel types of flow cytometers are emerging during the search to tackle these challenges. Notwithstanding the relevance of other promising alternatives to standard optical flow cytometry (e.g., mass cytometry, various optical and electrical microcytometers), this report focuses on a recent microcytometric technology based on magnetic sensors and magnetic particles integrated into microfluidic structures for dynamic bioanalysis of fluid samples-magnetic flow cytometry. Its concept, main developments, targeted applications, as well as the challenges and trends behind this technology are presented and discussed. Graphical abstract ᅟ "Kindly advise whether there is online abstract figure for this paper. If so, kindly resupply.The graphical abstract is correctly supplied.

Assessment of nanofiltration and reverse osmosis potentialities to recover metals, sulfuric acid, and recycled water from acid gold mining effluent.

This work assessed the potential of nanofiltration (NF) and reverse osmosis (RO) to treat acid streams contaminated with metals, such as effluent from the pressure oxidation process (POX) used in refractory gold ore processing. NF and RO were evaluated in terms of rejections of sulfuric acid and metals. Regarding NF, high sulfuric acid permeation (∼100%), was observed, while metals were retained with high efficiencies (∼90%), whereas RO led to high acid rejections (<88%) when conducted in pH values higher than 1. Thus, sequential use of NF and RO was proved to be a promising treatment for sulfuric acid solutions contaminated by metals, such as POX effluent. In this context, a purified acid stream could be recovered in NF permeate, which could be further concentrated in RO. Recovered acid stream could be reused in the gold ore processing or commercialized. A metal-enriched stream could be also recovered in NF retentate and transferred to a subsequent metal recovery stage. In addition, considering the high acid rejection obtained through the proposed system, RO permeate could be used as recycling water.

Capture and detection of DNA hybrids on paper via the anchoring of antibodies with fusions of carbohydrate binding modules and ZZ-domains.

Microfluidic paper-based analytical devices (µPADs) fabricated by wax-printing are suitable platforms for the development of simple and affordable molecular diagnostic assays for infectious diseases, especially in resource-limited settings. Paper devices can be modified for biological assays by adding appropriate reagents to the test areas. For this purpose, the use of affinity immobilization strategies can be a good solution for bioactive paper fabrication. This paper describes a methodology to capture labeled-DNA strands and hybrids on paper via the anchoring of antibodies with a fusion protein that combines a family 3 carbohydrate binding module (CBM) from Clostridium thermocellum, with high affinity to cellulose, and the ZZ fragment of the staphyloccocal protein A, which recognizes IgG antibodies via their Fc portion. Antibodies immobilized via CBM-ZZ were able to capture appropriately labeled (biotin, fluorescein) DNA strands and DNA hybrids. The ability of an antibody specific to biotin to discriminate complementary from noncomplementary, biotin-labeled targets was demonstrated in both spot and microchannel assays. Hybridization was detected by fluorescence emission of the fluorescein-labeled DNA probe. The efficiency of the capture of labeled-DNA by antibodies immobilized on paper via the CBM-ZZ construct was significantly higher when compared with a physical adsorption method where antibodies were simply spotted on paper without the intermediation of other molecules. The experimental proof of concept of wax-printed µPADs functionalized with CBM-ZZ for DNA detection at room temperature presented in this study constitutes an important step toward the development of easy to use and affordable molecular diagnostic tests.

Portuguese Global Medicines Access Index 2021: An Indicator to Measure Access to Hospital Medicines.

OBJECTIVES: Access to innovative and effective medication is a citizen's right. The main objectives of this study were to build an indicator to measure access to medicines within hospitals, the Global Medicines Access Index, and to identify the main existing barriers. METHODS: Cross-sectional study carried out in Portuguese National Health Service hospitals. A consensus methodology (expert panel of 7 members) was used to define which dimensions should be included in the index and the weighting that each should take. The panel identified 6 dimensions: access to innovative medicines, proximity distribution, shortages, access to medicines before financing decision, value-based healthcare, and access to medication depending on cost/funding. Data were collected through an electronic questionnaire (September 2021). RESULTS: The response rate was 61.2%. Most hospitals used medicines with and without marketing authorization before the funding decision. Monitoring and generating evidence of new therapies results is still insufficient. The identified barriers were the administrative burden as the major barrier in purchasing medicines, with a relevant impact on shortages of medicines. Most respondents (87%) had a proximity distribution program, mainly implemented in the pandemic context, and the price/funding model was only identified by 10% as a barrier to access. The 2021 Global Medicines Access Index was 66%. Shortages and value-based use of medicines were the dimensions that had more influence in lowering the index value. CONCLUSIONS: The new formula used to obtain a unique and multidimensional index for access to hospital medicines seems to be more sensitive and objective and will be used to monitor access.

Artificial intelligence integration in the drug lifecycle and in regulatory science: policy implications, challenges and opportunities.

Artificial intelligence tools promise transformative impacts in drug development. Regulatory agencies face challenges in integrating AI while ensuring reliability and safety in clinical trial approvals, drug marketing authorizations, and post-market surveillance. Incorporating these technologies into the existing regulatory framework and agency practices poses notable challenges, particularly in evaluating the data and models employed for these purposes. Rapid adaptation of regulations and internal processes is essential for agencies to keep pace with innovation, though achieving this requires collective stakeholder collaboration. This article thus delves into the need for adaptations of regulations throughout the drug development lifecycle, as well as the utilization of AI within internal processes of medicine agencies.

Decreased expression of S100A8/A9 in V30M related ATTRv amyloidosis.

INTRODUCTION: Hereditary Transthyretin Amyloidosis is a rare, progressive and life-threatening systemic disease with predominant peripheral and autonomic nervous system involvement caused by mutation of the transthyretin protein. The most common TTR mutation regarding to ATTRv is a substitution of a Methionine for a Valine at position 30 that predisposes TTR to form aggregates and fibrils. METHODS: S100A8 protein levels were measured in plasma samples from ATTRV30M patients and healthy donors. Additionally, S100A8/9 levels were measured in Schwann cells after incubation with human WT or V30M TTR. Moreover, bone marrow derived macrophages of either genetic background were generated and the expression of S100A8/9 was measured in response to toll like receptors agonists. RESULTS: S100A8/A9 mRNA levels are decreased in HSF V30M mice as compared with the WT. Moreover, S100A8 protein levels were found downregulated in plasma samples from ATTRV30M patients. Furthermore, we provide evidence for a dysregulated S100 expression by Schwann cells in response to TTRV30M and by mutated macrophages in response to toll like receptors agonists. CONCLUSION: The presence of TTRV30M impacts S100 expression, possibly contributing to the impaired immune activation of Schwann cells in nerves from ATTRV30M patients. This may be linked to the diminished immune cellular infiltration in these nerves, contributing in this way for the neuronal dysfunction present in the disease.

Disclosing transcriptomics network-based signatures of glioma heterogeneity using sparse methods.

Gliomas are primary malignant brain tumors with poor survival and high resistance to available treatments. Improving the molecular understanding of glioma and disclosing novel biomarkers of tumor development and progression could help to find novel targeted therapies for this type of cancer. Public databases such as The Cancer Genome Atlas (TCGA) provide an invaluable source of molecular information on cancer tissues. Machine learning tools show promise in dealing with the high dimension of omics data and extracting relevant information from it. In this work, network inference and clustering methods, namely Joint Graphical lasso and Robust Sparse K-means Clustering, were applied to RNA-sequencing data from TCGA glioma patients to identify shared and distinct gene networks among different types of glioma (glioblastoma, astrocytoma, and oligodendroglioma) and disclose new patient groups and the relevant genes behind groups' separation. The results obtained suggest that astrocytoma and oligodendroglioma have more similarities compared with glioblastoma, highlighting the molecular differences between glioblastoma and the others glioma subtypes. After a comprehensive literature search on the relevant genes pointed our from our analysis, we identified potential candidates for biomarkers of glioma. Further molecular validation of these genes is encouraged to understand their potential role in diagnosis and in the design of novel therapies.

Experience and Learning from the COVID-19 Pandemic in Portugal: Perceptions of Community Pharmacy Professionals.

Background: During the COVID-19 pandemic, community pharmacy (CP) professionals were among those who experienced the greatest risk of contracting SARS-CoV-2, which forced major adaptations. Objectives: The objectives of the study were to describe the changes implemented in CP professionals during the pandemic, understand the perception of professionals about their experience, and explore changes to remain. Methods: An observational and cross-sectional study was conducted via an online questionnaire (June-September 2020). The target population was CP professionals working in Portugal for >2 years and serving the public during the pandemic. Results: Of a total of 353 participants, 84% were female (mean age of 37.6 years), and 81% were pharmacists (mean professional experience of 12.9 years). In the management and organizational dimensions, the most mentioned changes were adaptation to legislative changes (90%), fluctuations in the treasury (82%), and reduction of working hours (46%). Only 2% resorted to simplified layoff. In the back office, there was a need to adapt stock management (93%) and purchase personal protective equipment (99%). In the front office, there was a change in service policies - wicket or conditional opening (92%), routes of the arrival of user requests (91%), and home delivery (82%). Physical changes occurred in 100% of pharmacies. The most frequently implemented procedures were the use of protection systems and PPE, articulation with hospital pharmacies for dispensing in proximity (75%), and training in this area (55%). Regarding interpersonal climate, improvements in the connection between team members are evident: increase in mutual help (57%), solidarity (54%), and group cohesion (50%); in the relationship with clients, the majority indicated the replacement of the usual user by third parties (71%), and changes in communication channels (increase in use of technological means 68%). Conclusions: Results illustrate the profound impact of the pandemic on CP professionals, both professionally and personally. It also highlights the importance of their roles in proximity and community support.

Introdução: Durante a pandemia de COVID-19, os profissionais de farmácia comunitária (FC) estiveram entre os que apresentaram maior risco de contrair SARS-CoV-2, o que obrigou a grandes adaptações. Objetivos: Descrever as alterações implementadas nas FC durante a pandemia, compreender a percepção dos profissionais sobre as suas vivências e explorar as mudanças a serem mantidas. Metodologia: estudo observacional e transversal (junho-setembro de 2020). A população alvo foram os profissionais de FC a trabalhar em Portugal há >2 anos e atender o público durante a pandemia. Resultados: 353 participantes, 84% do sexo feminino (idade média - 37,6 anos) e 81% eram farmacêuticos (média de experiência profissional de 12,9 anos). Nas dimensões "gestão e organização", as mudanças mais referidas foram a adaptação a alterações legislativas (90%), flutuações de tesouraria (82%) e redução do horário de trabalho (46%). Apenas 2% recorreram ao lay-off simplificado. No back office: necessidade de adequação do stock (93%) e aquisição de equipamentos de proteção individual (99%). No front office: alteração das políticas de atendimento ­ atendimento ao postigo ou abertura condicional (92%), vias de chegada dos pedidos dos utentes (91%) e entrega ao domicílio (82%). Alterações físicas ocorreram em 100% das farmácias. Os procedimentos implementados com maior frequência foram a utilização de sistemas de proteção e EPI, a articulação com farmácias hospitalares para dispensa de medicamentos de proximidade (75%) e formação nesta área (55%). Em relação ao clima interpessoal, foram evidentes as melhorias na ligação entre os membros da equipa: aumento da inter-ajuda (57%), solidariedade (54%) e coesão do grupo (50%); no relacionamento com os utentes, a maioria referiu a substituição do utente habitual por terceiros (71%) e alterações nos canais de comunicação (aumento da utilização de meios tecnológicos 68%). Conclusões: Os resultados ilustram o profundo impacto da pandemia nos profissionais de FC, tanto a nível profissional como pessoal. Também de destacar a importância do papel da FC como espaço de saúde de proximidade e apoio à comunidade.

Frequency-dependent stimulated and post-stimulated voltage control of magnetism in transition metal nitrides: towards brain-inspired magneto-ionics.

Magneto-ionics, which deals with the change of magnetic properties through voltage-driven ion migration, is expected to be one of the emerging technologies to develop energy-efficient spintronics. While a precise modulation of magnetism is achieved when voltage is applied, much more uncontrolled is the spontaneous evolution of magneto-ionic systems upon removing the electric stimuli (i.e., post-stimulated behavior). Here, we demonstrate a voltage-controllable N ion accumulation effect at the outer surface of CoN films adjacent to a liquid electrolyte, which allows for the control of magneto-ionic properties both during and after voltage pulse actuation (i.e., stimulated and post-stimulated behavior, respectively). This effect, which takes place when the CoN film thickness is below 50 nm and the voltage pulse frequency is at least 100 Hz, is based on the trade-off between generation (voltage ON) and partial depletion (voltage OFF) of ferromagnetism in CoN by magneto-ionics. This novel effect may open opportunities for new neuromorphic computing functions, such as post-stimulated neural learning under deep sleep.

Customizable Serious Speech Therapy Games with Dynamic Difficulty Adjustment for Children with Sigmatism.

Children with speech sound disorders should attend speech and language therapy and should practice the speech exercises regularly to surpass their speech difficulties. Since doing the speech exercises often may be tedious, there is the need to motivate children to practice them. During the COVID-19 pandemic, speech and language pathologists had the need to adapt their procedures to others with less physical contact. Here, we propose two serious games to motivate children with sigmatism on doing the speech exercises, which can be used at home and during face-to-face and online speech therapy sessions. The games use automatic speech recognition to classify speech productions. Visual and auditory feedback are used to help children understand their performance, and a hint system is used to help them perform the exercises correctly. A dynamic difficulty adjustment system is used to change the level of difficulty according to the child's speech performance in previous trials.

Construction of HER2-Specific HIV-1-Based VLPs.

Virus-like particles (VLPs) are nanoplatforms comprised of one or more viral proteins with the capacity to self-assemble without viral genetic material. VLPs arise as promising nanoparticles (NPs) that can be exploited as vaccines, as drug delivery vehicles or as carriers of imaging agents. Engineered antibody constructs, namely single-chain variable fragments (scFv), have been explored as relevant molecules to direct NPs to their target. A vector containing the scFv of an antibody, aimed at the human epidermal growth factor receptor 2 (HER2) and fused to the human immunodeficiency virus (HIV) protein gp41, was previously constructed. The work herein describes the early results concerning the production and the characterization of HIV-1-based VLPs expressing this protein, which could function as potential non-toxic tools for transporting drugs and/or imaging agents.