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METHODS: Retrospective cohort study with review of medical records of women assisted between 2015 and 2020. The variables were socio-demographic and SV characteristics, gestational age, reactions towards pregnancy and outcome. We compared outcome groups using the chi-square test, Fisher's exact test and the Kruskal-Wallis test. The significance level was 5%. RESULTS: We evaluated the medical records of 235 women, of which 153(65%) had undergone to abortion; 17(7.2%) had a spontaneous abortion; 19(8%) remained pregnant; 25(10.6%) had an abortion denied; and 21(8.9%) had been lost to follow-up. Out of the total number of women, 44(18.7%) were adolescents, 152(65.2%) were white and 201(88.5%) had an education ≥9 years. Women who remained pregnant had a known aggressor, disclosed the pregnancy (p < 0.001) and were more ambivalent (p < 0.001) than the other groups. Gestational age was higher in the denied abortion group than in the performed abortion group (p < 0.001). CONCLUSION: Feelings related to decision-making about abortion affected all groups, with differences. It is important to give women space to be heard, so they can make their own decisions.

Abortion care is possible in places with restrictive laws; however, women with more vulnerable characteristics did not seek the service. Legal restrictions interfere with women's decision-making about abortion and can promote inequality in gaining access to health services.

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The COVID-19 pandemic has resulted in millions of deaths globally, and while several diagnostic systems were proposed, real-time reverse transcription polymerase chain reaction (RT-PCR) remains the gold standard. However, diagnostic reagents, including enzymes used in RT-PCR, are subject to centralized production models and intellectual property restrictions, which present a challenge for less developed countries. With the aim of generating a standardized One-Step open RT-qPCR protocol to detect SARS-CoV-2 RNA in clinical samples, we purified and tested recombinant enzymes and a non-proprietary buffer. The protocol utilized M-MLV RT and Taq DNA pol enzymes to perform a Taqman probe-based assay. Synthetic RNA samples were used to validate the One-Step RT-qPCR components, demonstrating sensitivity comparable to a commercial kit routinely employed in clinical settings for patient diagnosis. Further evaluation on 40 clinical samples (20 positive and 20 negative) confirmed its comparable diagnostic accuracy. This study represents a proof of concept for an open approach to developing diagnostic kits for viral infections and diseases, which could provide a cost-effective and accessible solution for less developed countries.

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OBJECTIVE: To assess menstrual hygiene management (MHM) and period poverty in a group of low- and medium-income menstruating people. METHODS: A cross-sectional study was conducted at the University of Campinas, Brazil, from January to June 2022 with 535 participants aged 18-49. For data collection, we used the abbreviated version of the WHO Quality-of-Life questionnaire (WHOQOL-BREF) and a pre-tested questionnaire with items regarding MHM and period poverty. RESULTS: We found an inverse relationship between the risk of no access to supplies to perform menstrual hygiene, which increased by 2.5% and each point less in the environmental domain of quality of life (QoL), and this risk increased by up to 3.1 times among participants who reported difficulty making ends meet. The risk of not having adequate conditions to perform menstrual hygiene at home increased by up to 2.6% for every one-point drop in the environmental domain of QoL. The risk of not having adequate conditions to perform menstrual hygiene outside the home increased by up to 1.3% for less points in the psychological domain of QoL, and 44.4% of those who indicated inadequate conditions for menstrual hygiene reported two or more childbirths. CONCLUSIONS: Menstruating people who have an increased risk of lacking menstrual supplies and have an increased risk of inadequate conditions for MHM at home and outside the home, as well as those who have two or more deliveries and those having difficulties making ends meet, scored low on the QoL, especially in the environmental and psychological dimensions.

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AIMS: To evaluate the antifungal and antibiofilm activity of gallic acid derivatives TPP+-C10 and TPP+-C12 and their effects on mitochondrial function on two Candida albicans reference strains (ATCC 90029 and ATCC 10231). METHODS AND RESULTS: First, we determined minimal inhibitory concentration (MIC) using a microdilution assay. Both compounds exerted antifungal effects, and their MICs ranged from 3.9 to 13 µM, with no statistically significant differences between them (P > 0.05, t-test). These concentrations served as references for following assays. Subsequently, we measured oxygen consumption with a Clark electrode. Our observations revealed that both drugs inhibited oxygen consumption in both strains with TPP+-C12 exerting a more pronounced inhibitory effect. We then employed flow cytometry with TMRE as a probe to assess mitochondrial membrane potential. For each strain assayed, the compounds induced a decay in transmembrane potential by 75%-90% compared to the control condition (P < 0.05, ANOVA). Then, we measured ATP levels using a commercial kit. TPP+-C12 showed a 50% decrease of ATP content (P < 0.05 ANOVA), while TPP+-C10 exhibited a less pronounced effect. Finally, we assessed the antibiofilm effect using the MTT reduction assay. Both compounds were effective, but TPP+-C12 displayed a greater potency, requiring a lower concentration to inhibit 50% of biofilms viability (P < 0.05, t-test). CONCLUSIONS: Derivatives of gallic acid linked to a TPP+ group exert antifungal and antibiofilm activity through impairment of mitochondrial function in C. albicans.

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Acute leukemias (AL) are aggressive neoplasms with high mortality rates. Metabolomics and oxidative status have emerged as important tools to identify new biomarkers with clinical utility. To identify the metabolic differences between healthy individuals (HI) and patients with AL, a multiplatform untargeted metabolomic and lipidomic approach was conducted using liquid and gas chromatography coupled with quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS or GC-QTOF-MS). Additionally, the total antioxidant capacity (TAC) was measured. A total of 20 peripheral blood plasma samples were obtained from patients with AL and 18 samples from HI. Our analysis revealed 135 differentially altered metabolites in the patients belonging to 12 chemical classes; likewise, the metabolic pathways of glycerolipids and sphingolipids were the most affected in the patients. A decrease in the TAC of the patients with respect to the HI was evident. This study conducted with a cohort of Colombian patients is consistent with observations from other research studies that suggest dysregulation of lipid compounds. Furthermore, metabolic differences between patients and HI appear to be independent of lifestyle, race, or geographic location, providing valuable information for future advancements in understanding the disease and developing more global therapies.

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BACKGROUND: Lung cancer is the leading cause of cancer death worldwide. It has been reported that genetic and epigenetic factors play a crucial role in the onset and evolution of lung cancer. Previous reports have shown that essential transcription factors in embryonic development contribute to this pathology. Runt-related transcription factor (RUNX) proteins belong to a family of master regulators of embryonic developmental programs. Specifically, RUNX2 is the master transcription factor (TF) of osteoblastic differentiation, and it can be involved in pathological conditions such as prostate, thyroid, and lung cancer by regulating apoptosis and mesenchymal-epithelial transition processes. In this paper, we identified TALAM1 (Metastasis Associated Lung Adenocarcinoma Transcript 1) as a genetic target of the RUNX2 TF in lung cancer and then performed functional validation of the main findings. METHODS: We performed ChIP-seq analysis of tumor samples from a patient diagnosed with lung adenocarcinoma to evaluate the target genes of the RUNX2 TF. In addition, we performed shRNA-mediated knockdown of RUNX2 in this lung adenocarcinoma cell line to confirm the regulatory role of RUNX2 in TALAM1 expression. RESULTS: We observed RUNX2 overexpression in cell lines and primary cultured lung cancer cells. Interestingly, we found that lncRNA TALAM1 was a target of RUNX2 and that RUNX2 exerted a negative regulatory effect on TALAM1 transcription.

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The dysbiosis of the oral microbiome and vascular translocation of the periodontopathic microorganism to peripheral blood can cause local and systemic extra-oral inflammation. Microorganisms associated with the subgingival biofilm are readily translocated to the peripheral circulation, generating bacteremia and endotoxemia, increasing the inflammation in the vascular endothelium and resulting in endothelial dysfunction. This review aimed to demonstrate how the dysbiosis of the oral microbiome and the translocation of oral pathogen-induced inflammation to peripheral blood may be linked to cardiovascular diseases (CVDs). The dysbiosis of the oral microbiome can regulate blood pressure and activate endothelial dysfunction. Similarly, the passage of periodontal microorganisms into the peripheral circulation and their virulence factors have been associated with a vascular compartment with a great capacity to activate endothelial cells, monocytes, macrophages, and plaquettes and increase interleukin and chemokine secretion, as well as oxidative stress. This inflammatory process is related to atherosclerosis, hypertension, thrombosis, and stroke. Therefore, oral diseases could be involved in CVDs via inflammation. The preclinic and clinical evidence suggests that periodontal disease increases the proinflammatory markers associated with endothelial dysfunction. Likewise, the evidence from clinical studies of periodontal treatment in the long term evidenced the reduction of these markers and improved overall health in patients with CVDs.

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Several hydrolases have been described to degrade polyethylene terephthalate (PET) at moderate temperatures ranging from 25°C to 40°C. These mesophilic PET hydrolases (PETases) are less efficient in degrading this plastic polymer than their thermophilic homologs and have, therefore, been the subject of many protein engineering campaigns. However, enhancing their enzymatic activity through rational design or directed evolution poses a formidable challenge due to the need for exploring a large number of mutations. Additionally, evaluating the improvements in both activity and stability requires screening numerous variants, either individually or using high-throughput screening methods. Here, we utilize instead the design of chimeras as a protein engineering strategy to increase the activity and stability of Mors1, an Antarctic PETase active at 25°C. First, we obtained the crystal structure of Mors1 at 1.6 Å resolution, which we used as a scaffold for structure- and sequence-based chimeric design. Then, we designed a Mors1 chimera via loop exchange of a highly divergent active site loop from the thermophilic leaf-branch compost cutinase (LCC) into the equivalent region in Mors1. After restitution of an active site disulfide bond into this chimera, the enzyme exhibited a shift in optimal temperature for activity to 45°C and an increase in fivefold in PET hydrolysis when compared with wild-type Mors1 at 25°C. Our results serve as a proof of concept of the utility of chimeric design to further improve the activity and stability of PETases active at moderate temperatures.

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We used a mixed design study to analyze the inequalities and inequities in Maternal Mortality (MM) for Chocó (Colombia) between 2010-2018. The quantitative component consisted of an analytical ecological design, where proportions, ratios, measures of central tendency and rates ratios, rate difference, Gini and concentration indices were calculated to measure inequalities. The qualitative component had a phenomenological and interpretive approach. One hundred thirty-one women died in Choco between 2010-2018. The Maternal Mortality Ratio was 224/100.000 live births. The Gini coefficient was 0.35, indicating inequality in the distribution of the number of MM with respect to live births. The health service offers have been concentrated in the private sector in urban areas (77%). The exercise of midwifery has played an important role in maternal and perinatal care processes, especially in territories where the State has been absent. Nevertheless, it occurs in complex circumstances such as the armed conflict, lack of transportation routes, and income deficits, affecting the timelines and care quality for these vulnerable groups. MM in Chocó has been a consequence of deficiencies in the health system and weaknesses in its infrastructure (absence of a high level of maternal-perinatal care). This is in addition to the territory's geographical characteristics, which increase vulnerability and health risks for women and their newborns. In Colombia, as well as in other countries, many maternal and newborn deaths are preventable because their causes are due to social injustices.

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The increase in life expectancy has led to a higher incidence of osteoporosis, characterized by an imbalance in bone remodeling. Several drugs are used for its treatment, but most promote undesirable side effects. The present investigation evaluated the effects of two low concentrations of grape seed extract (GSE) rich in proanthocyanidins on MC3T3-E1 osteoblastic cells. The cells were cultured in an osteogenic medium and divided into control (C), 0.1 µg/mL GSE (GSE0.1), and 1.0 µg/mL GSE (GSE1.0) groups to evaluate cell morphology, adhesion, and proliferation, in situ alkaline phosphatase (ALP) detection, mineralization and immunolocalization of osteopontin (OPN). The data obtained were analyzed by statistical tests for a significance of 5%. Cell morphology was maintained with both GSE concentrations, whereas cell adhesion significantly increased within three days in all groups. Cell proliferation increased significantly at seven days of culture, followed by a significant decrease in all experimental periods, with no statistical difference among them. In situ detection of ALP and mineralization increased with time, but within each period, no statistical differences among groups were observed. The expression of osteopontin was distributed regularly with more intensity after 24 hours in the GSE0.1 group. After three days, OPN expression was more intense in the control group, followed by GSE0.1 and GSE1.0 groups. Data obtained suggest that low concentrations of GSE do not affect the morphology and may stimulate the functional activity of osteoblastic cells.

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Abstract The increase in life expectancy has led to a higher incidence of osteoporosis, characterized by an imbalance in bone remodeling. Several drugs are used for its treatment, but most promote undesirable side effects. The present investigation evaluated the effects of two low concentrations of grape seed extract (GSE) rich in proanthocyanidins on MC3T3-E1 osteoblastic cells. The cells were cultured in an osteogenic medium and divided into control (C), 0.1 µg/mL GSE (GSE0.1), and 1.0 µg/mL GSE (GSE1.0) groups to evaluate cell morphology, adhesion, and proliferation, in situ alkaline phosphatase (ALP) detection, mineralization and immunolocalization of osteopontin (OPN). The data obtained were analyzed by statistical tests for a significance of 5%. Cell morphology was maintained with both GSE concentrations, whereas cell adhesion significantly increased within three days in all groups. Cell proliferation increased significantly at seven days of culture, followed by a significant decrease in all experimental periods, with no statistical difference among them. In situ detection of ALP and mineralization increased with time, but within each period, no statistical differences among groups were observed. The expression of osteopontin was distributed regularly with more intensity after 24 hours in the GSE0.1 group. After three days, OPN expression was more intense in the control group, followed by GSE0.1 and GSE1.0 groups. Data obtained suggest that low concentrations of GSE do not affect the morphology and may stimulate the functional activity of osteoblastic cells.

Resumo O aumento da expectativa de vida tem levado a uma maior incidência de osteoporose, caracterizada por um desequilíbrio na remodelação óssea. Vários medicamentos são utilizados para o seu tratamento, contudo, a maioria promove efeitos colaterais indesejáveis. A presente investigação avaliou os efeitos de duas baixas concentrações de extrato de semente de uva (GSE) rico em proantocianidinas em células osteoblásticas MC3T3-E1. As células foram cultivadas em meio osteogênico e divididas em grupos controle (C), 0,1 µg/mL de GSE (GSE0.1) e 1,0 µg/mL de GSE (GSE1.0) para avaliar morfologia, adesão e proliferação celular, detecção in situ de fosfatase alcalina (ALP), mineralização e imunolocalização da proteína osteopontina (OPN). Os dados obtidos foram analisados por testes estatísticos para um nível de significância de 5%. A proliferação celular aumentou significativamente aos sete dias de cultura, seguido de uma diminuição significativa em todos os períodos experimentais, sem diferença estatística entre eles. A detecção in situ de ALP e mineralização aumentou com o tempo, mas dentro de cada período não foram observadas diferenças estatísticas entre os grupos. A morfologia celular foi mantida com ambas as concentrações de GSE, enquanto a adesão celular aumentou significativamente aos três dias em todos os grupos. A expressão de osteopontina distribuiu-se regularmente com maior intensidade após 24 horas no grupo GSE0.1. Após três dias, a expressão de OPN foi mais intensa no grupo controle, seguida pelos grupos GSE0.1 e GSE1.0. Os dados obtidos sugerem que baixas concentrações de GSE não afetam a morfologia e podem estimular a atividade funcional das células osteoblásticas.

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This article discusses the violence perpetrated by clients against social workers who provided services to COVID-19 patients and their families in Chilean hospitals during the initial peak of the disease in the country. In this way, this becomes the first study to analyze client violence toward social workers in the health crisis generated by the pandemic. The results indicate that the presence of aggressions from clients is high and social workers' coping strategies are passive. Finally, the urgent call for prevention and the need for research are presented.

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Hedgehog proteins (Hhs) secretion from apical and/or basolateral domains occurs in different epithelial cells impacting development and tissue homeostasis. Palmitoylation and cholesteroylation attach Hhs to membranes, and Dispatched-1 (Disp-1) promotes their release. How these lipidated proteins are handled by the complex secretory and endocytic pathways of polarized epithelial cells remains unknown. We show that polarized Madin-Darby canine kidney cells address newly synthesized sonic hedgehog (Shh) from the TGN to the basolateral cell surface and then to the apical domain through a transcytosis pathway that includes Rab11-apical recycling endosomes (Rab11-ARE). Both palmitoylation and cholesteroylation contribute to this sorting behavior, otherwise Shh lacking these lipid modifications is secreted unpolarized. Disp-1 mediates first basolateral secretion from the TGN and then transcytosis from Rab11-ARE. At the steady state, Shh predominates apically and can be basolaterally transcytosed. This Shh trafficking provides several steps for regulation and variation in different epithelia, subordinating the apical to the basolateral secretion.

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Resumen Antecedentes: la pandemia y el confinamiento por COVID-19 modificaron las condiciones de vida de los escolares y sus familias. Objetivo: describir la ejecución de una intervención educativa alimentaria en escolares de la Institución Educativa Monseñor Gerardo Valencia Cano, Medellín, Colombia, antes y durante el confinamiento por COVID-19. Materiales y métodos: investigación cualitativa con apoyo de variables cuantitativas. La recolección y análisis de datos se hicieron con base en la teoría fundamentada. La población estuvo conformada por escolares y padres de familia de la Institución Educativa Monseñor Gerardo Valencia Cano. Antes de la COVID-19 se realizaron cuatro grupos focales y una encuesta semiestructurada a 416 estudiantes y sus padres de familia, y con 42 docentes se realizaron cuatro grupos focales más; estos se realizaron hasta alcanzar la saturación. Durante el confinamiento se aplicaron 237 encuestas a familias (escolares y padres). Se evaluó el estado nutricional. Resultados: el 37 % de los escolares entre tercero y séptimo grado tenía exceso de peso. Se logró identificar de manera participativa los factores de riesgo y protectores frente al exceso de peso, para definir una intervención educativa alimentaria desde y con los sujetos. Los nodos conceptuales de la intervención fueron alimentación saludable, métodos de cocción, preparación de alimentos saludables y alimentación para la prevención del exceso de peso y cáncer de colon. Conclusiones: el reconocimiento de las condiciones alimentarias antes y después del confinamiento de los escolares y sus familias, o del cambio de estas a lo largo de la intervención educativa, contribuyó a lograr coherencia entre las necesidades, objetivos educativos y metodologías implementadas. El hecho de tener reunidos en un mismo espacio (hogar) a los escolares y sus padres durante el confinamiento propició un microentorno favorecedor de conductas que motivaron una mejora de los hábitos alimentarios familiares.

Abstract: Background: The pandemic and COVID-19 confinement changed the living conditions of schoolchildren and their families. Objective: To describe the implementation of an educational food intervention in schoolchildren at the Monseñor Gerardo Valencia Cano Educational Institution, Medellín, Colombia, before and during the COVID-19confinement. Materials and Methods: Qualitative research supported by quantitative variables. Data collection and analysis were based on grounded theory. The population consisted of schoolchildren and parents of the Monseñor Gerardo Valencia Cano Educational Institution. Before COVID-19, four focus groups and a semi-structured survey were conducted with 416 students and their parents, and four more focus groups were conducted with 42 teachers; these were carried out until saturation was reached. During the confinement, 237 surveys were administered to families (students and parents). Nutritional status was evaluated. Results: 37% of the students between third and seventh grade were overweight. It was possible to identify in a participatory way the risk and protective factors against excess weight, in order to define an educational food intervention from and with the subjects. The conceptual nodes of the intervention were healthy eating, cooking methods, preparation of healthy food, and food for the prevention of excess weight and colon cancer. Conclusions: The recognition of the nutritional conditions before and after the confinement of the schoolchildren and their families, or the change of these throughout the educational intervention, helped to achieve coherence between the needs, educational objectives, and methodologies implemented. The fact of having schoolchildren and their parents gathered in the same space (home) during confinement generated a microenvironment that favored behaviors that triggered an improvement in family eating habits.

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Objetivo: Presentar los resultados de una revisión sobre la investigación y metodología de Sistematización de Experiencias y su pertinencia en la salud pública. Métodos: Se realizó una revisión de literatura en dos etapas usando los siguientes operadores booleanos: "sistematización de experiencias" AND "metodología" OR "investigación" OR "salud" OR "educación popular" OR "trabajo social" OR "Latinoamérica" OR "Investigación Acción Participativa", en las bases de datos MedLine y Scielo, complementada con la búsqueda en el catálogo general de las bibliotecas de la Pontificia Universidad Javeriana y la Universidad Nacional de Colombia. Se dejó fecha e idioma abierto. Se empleó N-Vivo versión 12 para el procesamiento de la información. Resultados: Se obtuvieron 183 referencias, entre artículos, libros, literatura gris, tesis (pregrado, maestría o doctorado) y material audiovisual. De estos, se seleccionaron 88 para revisión de texto completo. Se generaron seis categorías relacionadas con la definición, metodología, instrumentos, objetivos, limites-tensiones y ventajas de la Sistematización de Experiencias. Conclusiones: La Sistematización de Experiencias es una metodología cualitativa surgida en el contexto latinoamericano que permite la reconstrucción de la historia y el conocimiento de las prácticas para fortalecer y transformar la realidad local. En el campo de la salud pública, la SE permite reinterpretar las nociones y problemáticas en salud de las personas y colectividades; comprender sus experiencias, necesidades y expectativas en salud; e identificar la agencia de las personas para mejorar e incidir en el bienestar y la calidad de vida de sus comunidades.

Objective: To show the results of a literature review about the investigation and methodology of Systematization of Experiences and its relevance in public health. Method: A literature review was carried out in two stages. The following Boolean operators were used: "systematization of experiences" AND "methodology" OR "research" OR "health" OR "popular education" OR "social work" OR "Latin America" OR "Participatory Action Research", in the MedLine and Scielo databases, was complemented by a search in the general catalog of the libraries of the Pontificia Universidad Javeriana and the Universidad Nacional de Colombia. Date and language were left open. N-Vivo version 12 was used for the processing and description of the information. Results: A total of 183 references were obtained from articles, books, gray literature, thesis (undergraduate, master's or doctorate) and audiovisual material. Of these, 88 were selected for full text review. Six categories were created related to the definition, methodology, instruments, objectives, limits-tensions and advantages of the Systematization of experiences. Conclusions: The Systematization of Experiences is a methodological strategy that contributes to qualitative research created within the context of Latin America, which allows us to recover the history and knowledge of practices while also strengthening and transforming the local reality. In the field of public health, it allows us to reinterpreting the notions and problems of individuals and communities; understanding their experiences, needs and expectations in health; and identify people's agency to improve and positively influence the well-being and quality of life of the community.

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Polyethylene terephthalate (PET) is one of the most widely used synthetic plastics in the packaging industry, and consequently has become one of the main components of plastic waste found in the environment. However, several microorganisms have been described to encode enzymes that catalyze the depolymerization of PET. While most known PET hydrolases are thermophilic and require reaction temperatures between 60°C and 70°C for an efficient hydrolysis of PET, a partial hydrolysis of amorphous PET at lower temperatures by the polyester hydrolase IsPETase from the mesophilic bacterium Ideonella sakaiensis has also been reported. We show that polyester hydrolases from the Antarctic bacteria Moraxella sp. strain TA144 (Mors1) and Oleispira antarctica RB-8 (OaCut) were able to hydrolyze the aliphatic polyester polycaprolactone as well as the aromatic polyester PET at a reaction temperature of 25°C. Mors1 caused a weight loss of amorphous PET films and thus constitutes a PET-degrading psychrophilic enzyme. Comparative modeling of Mors1 showed that the amino acid composition of its active site resembled both thermophilic and mesophilic PET hydrolases. Lastly, bioinformatic analysis of Antarctic metagenomic samples demonstrated that members of the Moraxellaceae family carry candidate genes coding for further potential psychrophilic PET hydrolases. IMPORTANCE A myriad of consumer products contains polyethylene terephthalate (PET), a plastic that has accumulated as waste in the environment due to its long-term stability and poor waste management. One promising solution is the enzymatic biodegradation of PET, with most known enzymes only catalyzing this process at high temperatures. Here, we bioinformatically identified and biochemically characterized an enzyme from an Antarctic organism that degrades PET at 25°C with similar efficiency to the few PET-degrading enzymes active at moderate temperatures. Reasoning that Antarctica harbors other PET-degrading enzymes, we analyzed available data from Antarctic metagenomic samples and successfully identified other potential enzymes. Our findings contribute to increasing the repertoire of known PET-degrading enzymes that are currently being considered as biocatalysts for the biological recycling of plastic waste.

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RT-LAMP (reverse transcription - Loop-mediated isothermal amplification) has gained popularity for the detection of SARS-CoV-2. The high specificity, sensitivity, simple protocols and potential to deliver results without the use of expensive equipment has made it an attractive alternative to RT-PCR. However, the high cost per reaction, the centralized manufacturing of required reagents and their distribution under cold chain shipping limits RT-LAMP's applicability in low-income settings. The preparation of assays using homebrew enzymes and buffers has emerged worldwide as a response to these limitations and potential shortages. Here, we describe the production of Moloney murine leukemia virus (M-MLV) Reverse Transcriptase and BstLF DNA polymerase for the local implementation of RT-LAMP reactions at low cost. These reagents compared favorably to commercial kits and optimum concentrations were defined in order to reduce time to threshold, increase ON/OFF range and minimize enzyme quantities per reaction. As a validation, we tested the performance of these reagents in the detection of SARS-CoV-2 from RNA extracted from clinical nasopharyngeal samples, obtaining high agreement between RT-LAMP and RT-PCR clinical results. The in-house preparation of these reactions results in an order of magnitude reduction in costs, and thus we provide protocols and DNA to enable the replication of these tests at other locations. These results contribute to the global effort of developing open and low cost diagnostics that enable technological autonomy and distributed capacities in viral surveillance.

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Natural substances with antioxidant effects may benefit prevention and treatment of people with or prone to bone diseases after menopause, such as osteoporosis. This study aimed to evaluate the in vitro effect of preadministration of yerba mate extract (YM) in the metabolism of MC3T3-E1 osteoblasts exposed to hydrogen peroxide (H2O2). The cells (MC3T3-E1) were cultured in 24-well plates with the concentration of 1 µg/mL yerba mate extract dissolved in culture medium throughout the culture period. Four hours before each experiment, 400 µmol/L H2O2 was added per well to simulate oxidative stress. There were evaluated cell adhesion and proliferation, in situ detection of alkaline phosphatase (ALP), mineralized nodules, and immunolocalization of osteocalcin (OCN), bone sialoprotein (BSP) and alkaline phosphatase (ALP) proteins. The results showed that YM preadministration to H2O2 exposition significatively increased cell adhesion after 3 days as well as proliferation and in situ ALP detection after 10 and 7 days respectively, when compared to H2O2 group. Besides, staining of OCN and BSP proteins was less intense and scattered in poor spread cells with cytoskeletal changes in H2O2 group when compared to control and YM H2O2 group. ALP staining was restrained to intracellular regions and similar in all experimental groups. Our results suggest that preadministration of yerba mate extract may prevent deleterious effects in the morphology and functional activity of osteoblasts exposed to H2O2, which could enable the maintenance of extracellular matrix in the presence of oxidative stress.

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BACKGROUND: Several drugs are capable of promoting changes in bone metabolism. The aim of this study was to evaluate the effect of long-term low-dose aspirin (LDA) therapy on implant osseointegration. METHODS: Male Wistar rats were divided into 4 groups (n = 8/group) according to oral gavage solution received prior (42 days) to the implant surgery on the tibia. The control group was treated with saline solution for 7 (CG-7) and 28 (CG-28) days. The use of low-dose aspirin was performed in AG groups (6.75 mg/kg of aspirin) for 7 (AG-7) and 28 (AG-28) days. After experimental periods, histomorphometric evaluation of bone-to-implant contact (BIC) and the bone area between threads (BABT) was performed. RESULTS: Reduced BIC values were detected in AG-7 (62.8% ± 17.1) group compared to AG-28 (91.9% ± 5.4), CG-7 (82.7% ± 15.2), and CG-28 (89.9% ± 9.7). BABT evaluation revealed lower values in AG-7 (70.9% ± 15.2) compared to AG-28 (95.4% ± 3.7) and CG-28 (87.1% ± 10.2) groups. CONCLUSIONS: The treatment with low doses of aspirin promoted a discrete inhibitory effect in the early stages (7 days) of repair after implant placement, specifically in the bone deposition. However, these effects were not detected in the late stages (28 days), considering BIC and BABT parameters.

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Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) has gained popularity for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The high specificity, sensitivity, simple protocols, and potential to deliver results without the use of expensive equipment has made it an attractive alternative to RT-PCR. However, the high cost per reaction, the centralized manufacturing of required reagents, and their distribution under cold chain shipping limit RT-LAMP's applicability in low-income settings. The preparation of assays using homebrew enzymes and buffers has emerged worldwide as a response to these limitations and potential shortages. Here, we describe the production of Moloney murine leukemia virus reverse transcriptase and BstLF DNA polymerase for the local implementation of RT-LAMP reactions at low cost. These reagents compared favorably to commercial kits, and optimum concentrations were defined in order to reduce time to threshold, increase ON/OFF range, and minimize enzyme quantities per reaction. As a validation, we tested the performance of these reagents in the detection of SARS-CoV-2 from RNA extracted from clinical nasopharyngeal samples, obtaining high agreement between RT-LAMP and RT-PCR clinical results. The in-house preparation of these reactions results in an order of magnitude reduction in costs; thus, we provide protocols and DNA to enable the replication of these tests at other locations. These results contribute to the global effort of developing open and low-cost diagnostics that enable technological autonomy and distributed capacities in viral surveillance.

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