Antimalarials and macrolides: a review of off-label pharmacotherapies during the first wave of the SARS-CoV-2 pandemic

We critically analyzed clinical trials performed with chloroquine (CQ) and hydroxychloroquine (HCQ) with or without macrolides during the first wave of COVID-19 and discussed the design and limitations of peer-reviewed studies from January to July 2020. Seventeen studies were eligible for the discussion. CQ and HCQ did not demonstrate clinical advantages that justified their inclusion in therapeutic regimens of free prescription for treatment or prophylactic purposes, as suggested by health authorities, including in Brazil, during the first wave. Around August 2020, robust data had already indicated that pharmacological effects of CQ, HCQ and macrolides as anti-SARS-CoV-2 molecules were limited to in vitro conditions and largely based on retrospective trials with low quality and weak internal validity, which made evidence superficial for decision-making. Up to that point, most randomized and nonrandomized clinical trials did not reveal beneficial effects of CQ or HCQ with or without macrolides to reduce lethality, rate of intubation, days of hospitalization, respiratory support/mechanical ventilation requirements, duration, type and number of symptoms, and death and were unsuccessful in increasing virus elimination and/or days alive in hospitalized or ambulatory patients with COVID-19. In addition, many studies have demonstrated that side effects are more common in CQ-or HCQ-treated patients

Efeito antimetastático e mecanismos de ação de proteases do látex de Vasconcellea cundinamarcensis (Carica candamarcensis)

Exportado OPUSMade available in DSpace on 2019-08-12T20:56:56Z (GMT). No. of bitstreams: 1 disserta__o_dalton___11032011.pdf: 358316 bytes, checksum: 1fedb416fc6abf49e6829e25de0bd524 (MD5) Previous issue date: 11A fração proteolítica P1G10 do látex de Vasconcellea undinamarcensis apresenta atividade antitumoral e/ou antimetastática em modelos murinos de melanoma e carcinoma de Ehrlich. O objetivo desse trabalho foi avaliar as sub-frações de P1G10, CMS-1 e CMS-2, quanto ao efeito antimetastático em carcinoma de cólon (CT26.WT) e melanoma murino (B16-F10) e possíveis mecanismos de ação. No carcinoma de cólon, 1 ou 5 mg/kg de P1G10 e 5 mg/kg de CMS-2 apresentaram efeito antimetastático e, sobre o melanoma, esse efeito foi observado com 5 mg/kg de CMS-2 e também com 5 mg/kg de P1G10, conforme trabalhos anteriores do grupo de pesquisa. P1G10 demonstrou citotoxicidade contra células normais (CHO e BHK-21) e tumorais (CT26.WT e B16-F10), com IC-50 iguais ou menores que 21 g/mL, enquanto que CMS-2 apresentou esse efeito seletivamente sobre células tumorais (IC-50 menor que 11 g/mL). P1G10 ou CMS-2 100 g/mL promoveu um aumento do conteúdo sub-diplóide das células expostas em diferentes períodos analisados. Ainda, P1G10 exerce esse efeito por uma ação dependente da sua atividade proteolítica e por uma via caspase-dependente. O tratamento com 100 g/mL de P1G10, 2h, promoveu uma diminuição na adesão celular das células tumorais aos componentes da matriz extracelular e ainda, na capacidade de invasão dessas células. Em todo o estudo, CMS-1 não demonstrou qualquer efeito significativo. Assim, as proteases contidas em CMS-2 são as responsáveis pela ação antimetastática de P1G10 e contribui para esse efeito a citotoxicidade, por apoptose e dependente de suas atividades proteolíticas e, ainda, a redução da adesão e invasão celular.The proteolytic fraction P1G10 from Vasconcellea cundinamarcensis latex shows antitumor and/or antimetastatic activity in murine melanoma and Ehrlich carcinoma. The aim of this study was to evaluate the P1G10 sub-fractions, CMS-1 and CMS-2, regarding the antimetastatic effect in murine colon carcinoma (CT26.WT) and melanoma (B16-F10) and its possible mechanisms of action. In colon carcinoma, 1 or 5 mg/kg of P1G10 and 5 mg/kg of CMS-2 showed antimetastatic effect. In melanoma, this effect was observed with 5 mg/kg of CMS-2 and with 5 mg/kg of P1G10, as previous studies by the research group. P1G10 showed cytotoxicity against normal (CHO and BHK-21) and tumor (B16-F10 and CT26.WT) cells, with IC-50 equal or less than 21 g/mL, while CMS-2 had this effect selectively on tumor cells (IC-50 less than 11 g/mL). CMS-2 or P1G10 100 g/mL lead to an increase in the sub-diploid DNA content in cells exposed to different periods. Still, P1G10 exerts this effect by an action dependent on its proteolytic activity and a caspase-dependent pathway. Treatment with P1G10 100 g/mL, 2h, promoted a decrease in tumor cells adhesion to extracellular matrix components and to the invasiveness of these cells. Throughout the study, CMS-1 showed no significant effect. Thus, the proteases contained in CMS-2 are responsible for the antimetastatic action of P1G10 and contributes to this effect the cytotoxicity, through apoptosis and dependent on their proteolytic activities, and also the reduction of cellular adhesion and invasio

Antiangiogenesis, Loss of Cell Adhesion and Apoptosis Are Involved in the Antitumoral Activity of Proteases from V. cundinamarcensis (C. candamarcensis) in Murine Melanoma B16F1

The proteolytic enzymes from V. cundinamarcensis latex, (P1G10), display healing activity in animal models following various types of lesions. P1G10 or the purified isoforms act as mitogens on fibroblast and epithelial cells by stimulating angiogenesis and wound healing in gastric and cutaneous ulcers models. Based on evidence that plant proteinases act as antitumorals, we verified this effect on a murine melanoma model. The antitumoral effect analyzed mice survival and tumor development after subcutaneous administration of P1G10 into C57BL/6J mice bearing B16F1 low metastatic melanoma. Possible factors involved in the antitumoral action were assessed, i.e., cytotoxicity, cell adhesion and apoptosis in vitro, haemoglobin (Hb), vascular endothelial growth factor (VEGF), tumor growth factor-β (TGF-β), tumor necrosis factor-α (TNF-α) content and N-acetyl-glucosaminidase (NAG) activity. We observed that P1G10 inhibited angiogenesis measured by the decline of Hb and VEGF within the tumor, and TGF-β displayed a non-significant increase and TNF-α showed a minor non-significant reduction. On the other hand, there was an increase in NAG activity. In treated B16F1 cells, apoptosis was induced along with decreased cell binding to extracellular matrix components (ECM) and anchorage, without impairing viability

Participação da proteinase CMS2MS3 na atividade antimetastática de fração do látex de Vasconcellea cundinamarcensis: efeitos sobre a adesão e morte celular

Exportado OPUSMade available in DSpace on 2019-08-13T18:49:01Z (GMT). No. of bitstreams: 1 tese___dalton_dittz.pdf: 1627783 bytes, checksum: 391892b14ee57305a76508886d1f69f6 (MD5) Previous issue date: 7Nosso grupo de pesquisa demonstrou que frações proteolíticas de látex Vasconcellea cundinamarcensis (P1G10 e CMS2) têm atividade antitumoral e antimetastática em melanoma murino com redução da adesão e migração celular e indução de apoptose. Quando CMS2 é submetida à cromatografia em resina Mono- S Sepharose (FPLC), cinco proteases (CMS2MS1-5) são isoladas. Nosso objetivo foi identificar a(s) protease(s) presentes nas frações menos purificadas, e avaliar aatividade na adesão e morte celular que contribuem para os efeitos previamente descritos na linhagem celular de melanoma murino metastático (B16F10). Dentre as cinco proteases analisadas, CMS2MS3 mostrou a menor CC-50 (7,81 g/mL) em comparação com as demais (34,5 - 303,5 g/mL). Todas as proteases, especialmente CMS2MS3 (10 - 50 g/mL), reduziram a adesão de B16F10 à placas de poliestireno. CMS2MS3 também promoveu uma redução na adesão de célulasB16F10 aos componentes da matriz extracelular, especialmente vitronectina e laminina, após o tratamento com 10 g/mL por 2, 4, 8 ou 24 h. O nível das subunidades 4, 5 e 1 de integrina foi reduzido por CMS2MS3 10 g/mL em todos os tempos analisados, mostrando que este efeito contribui, pelo menos parcialmente, para a perda da adesão. A quantidade de vinculina/célula reduziu em 65-85% a partir de 2h de exposição à CMS2MS3, mostrando que a adesão focal pode estar ligada às integrinas composta pelas subunidades 4, 5 ou 1. Após 2 -24h de exposição à CMS2MS3, a quantidade de DNA sub-diplóide aumentou apenas para a concentração de 50 g/mL, embora, na concentração de 10 g/mL, já se observa uma redução nos níveis intracelulares de pró-caspases 3 e 9, a partir de 2h de exposição, mostrando que a morte celular ocorre por apoptose e após a perdada adesão. Observou-se, também, um aumento rápido do cálcio nuclear e mitocondrial após a exposição de B16F10 à CMS2MS3 10 g/mL, o que pode estar relacionado à liberação de citocromo c e ativação da apoptose. Assim, dentre as cinco proteases de CMS2, CMS2MS3 apresentou o melhor efeito citotóxico e capacidade de reduzir a adesão, mediada pelas subunidades 4, 5 e 1 de integrinas, nas células B16F10. A morte celular, por apoptose, ocorre após a perda de adesão e, associado a este último efeito, contribui para reduzir a capacidade decolonização pulmonar.Our research group has demonstrated that proteolytic fractions P1G10 and CMS2 (derived from P1G10) of Vasconcellea cundinamarcensis latex display antitumor and antimetastatic activity on murine melanoma as demonstrated by reduction in cell adhesion, migration, and induction of apoptosis. The CMS2 was further resolved into five proteases (CMS2MS1-5) when chromatographed on Mono-S Sepharose resin(FPLC). These five proteases were analyzed for their ability to promote cell adhesion changes and death effects seen earlier in less purified fractions, by using a murine metastatic melanoma cell line (B16F10). Among the five analyzed proteases, CMS2MS3 showed the lowest CC-50 (7.81 g/mL) compared to others (34.5 to 303.5 g/mL). Each protease reduced B16F10 adhesion on polystyrene plates, particularly CMS2MS3 at 10 - 50 g/mL. This protease also promoted a reduction in the B16F10 cell adhesion to extracellular matrix components, especially towardsvitronectin and laminin, after treatment with 10 g/mL for 2, 4, 8 or 24 h. The level of 4, 5 and 1 subunits of integrin was reduced when cells were exposed to 10 g/mL CMS2MS3 at each interval analyzed demonstrating that this effect contributes, at least partially, with the loss of adhesion. The number of vinculin/cell reduced 65 85% after 2 h exposure to CMS2MS3, showing that focal adhesion may be linked tothat integrin composed of subunits 4, 5 and 1. After 2 24 h exposure to 50 g/mL CMS2MS3, sub-diploid DNA was increased. However, in lower concentration (10 ug/mL) CMS2MS3 induced a reduction in intracellular levels of pro-caspases 3 and 9 from 2h of exposure of cells, suggesting that cell death ensues apoptosis and loss of cell adhesion. A rapid increase in nuclear and mitochondrial calcium was observedafter B16F10 exposure to CMS2MS3 10 g/mL and this may be related to the release of cytochrome c and activation of apoptosis. Thus, CMS2MS3 showed the strongest cytotoxic effect and the ability to reduce cell adhesion, mediated by 4, 5 and 1 subunits of integrin in B16F10 cells. The cell death observed is probably by apoptosis, after loss of adhesion and might contributes to impair the ability of tumorlung colonization

Cysteine Proteases from V. cundinamarcensis (C. candamarcensis) Inhibit Melanoma Metastasis and Modulate Expression of Proteins Related to Proliferation, Migration and Differentiation

Previous studies showed that P1G10, a proteolytic fraction from Vasconcellea cundinamarcensis latex, reduced the tumor mass in animals bearing melanoma, increased in vitro DNA fragmentation and decreased cell adhesion. Here, we present some molecular and cellular events related to the antimetastatic effect induced by the CMS-2 fraction derived from P1G10 in metastatic melanoma B16-F10 and melanocyte Melan-a. Using difference gel electrophoresis and mass spectrometry, we identified four proteins overexpressed in tumor cells, all of them related to proliferation, survival, migration and cell invasion, that had their expression normalized upon treatment with CMS-2: nucleophosmin 1, heat shock protein 65, calcyclin binding protein and eukaryotic translation initiation factor 4H. In addition, some antioxidant and glycolytic enzymes show increased expression after exposure to CMS-2, along with an induction of melanogenesis (differentiation marker). The down regulation of cofilin 1, a protein involved in cell motility, may explain the inhibition of cell migration and dendritic-like outgrowth in B16-F10 and Melan-a, observed after CMS-2 treatment. Taken together, it is argued that CMS-2 modulates the expression of proteins related to metastatic development, driving the cell to a more differentiated-like state. These effects support the CMS-2 antimetastatic activity and place this fraction in the category of anticancer agent

Antiangiogenesis, Loss of Cell Adhesion and Apoptosis Are Involved in the Antitumoral Activity of Proteases from V. cundinamarcensis (C. candamarcensis) in Murine Melanoma B16F1

The proteolytic enzymes from V. cundinamarcensis latex, (P1G10), display healing activity in animal models following various types of lesions. P1G10 or the purified isoforms act as mitogens on fibroblast and epithelial cells by stimulating angiogenesis and wound healing in gastric and cutaneous ulcers models. Based on evidence that plant proteinases act as antitumorals, we verified this effect on a murine melanoma model. The antitumoral effect analyzed mice survival and tumor development after subcutaneous administration of P1G10 into C57BL/6J mice bearing B16F1 low metastatic melanoma. Possible factors involved in the antitumoral action were assessed, i.e., cytotoxicity, cell adhesion and apoptosis in vitro, haemoglobin (Hb), vascular endothelial growth factor (VEGF), tumor growth factor-β (TGF-β), tumor necrosis factor-α (TNF-α) content and N-acetyl-glucosaminidase (NAG) activity. We observed that P1G10 inhibited angiogenesis measured by the decline of Hb and VEGF within the tumor, and TGF-β displayed a non-significant increase and TNF-α showed a minor non-significant reduction. On the other hand, there was an increase in NAG activity. In treated B16F1 cells, apoptosis was induced along with decreased cell binding to extracellular matrix components (ECM) and anchorage, without impairing viability

The Pollutant Organotins Leads to Respiratory Disease by Inflammation: A Mini-Review

Organotins (OTs) are organometallic pollutants. The OTs are organometallic pollutants that are used in many industrial, agricultural, and domestic products, and it works as powerful biocidal compound against large types of microorganisms such as fungi and bacteria. In addition, OTs are well known to be endocrine-disrupting chemicals, leading abnormalities an ?imposex? phenomenon in the female mollusks. There are some studies showing that OTs? exposure is responsible for neural, endocrine, and reproductive dysfunctions in vitro and in vivo models. However, OTs? effects over the mammalian immune system are poorly understood, particularly in respiratory diseases. The immune system, as well as their cellular components, performs a pivotal role in the control of the several physiologic functions, and in the maintenance and recovery of homeostasis. Thus, it is becoming important to better understand the association between environmental contaminants, as OTs, and the physiological function of immune system. There are no many scientific works studying the relationship between OTs and respiratory disease, especially about immune system activation. Herein, we reported studies in animal, humans, and in vitro models. We searched studies in PUBMED, LILACS, and Scielo platforms. Studies have reported that OTs exposure was able to suppress T helper 1 (Th1) and exacerbate T helper 2 (Th2) response in the immune system. In addition, OTs? contact could elevate in the airway inflammatory response, throughout a mechanism associated with the apoptosis of T-regulatory cells and increased oxidative stress response. In addition, OTs induce macrophage recruitment to the tissue, leading to the increased necrosis, which stimulates an inflammatory cytokines secretion exacerbating the local inflammation and tissue function loss. Thus, the main intention of this mini-review is to up to date the main findings involving the inflammatory profile (especially Th1 and Th2 response) in the respiratory tract as a result of OTs? exposure

Nanoparticles Obtained from Zein for Encapsulation of Mesalazine

We encapsulated MSZ in zein nanoparticles (NP-ZN) using a desolvation method followed by drying in a mini spray dryer. These nanoparticles exhibited a size of 266.6 ± 52 nm, IPD of 0.14 ± 1.1 and zeta potential of −36.4 ± 1.5 mV, suggesting colloidal stability. Quantification using HPLC showed a drug-loaded of 43.8 µg/mg. SEM demonstrated a spherical morphology with a size variation from 220 to 400 nm. A FTIR analysis did not show drug spectra in the NPs in relation to the physical mixture, which suggests drug encapsulation without changing its chemical structure. A TGA analysis showed thermal stability up to 300 °C. In vitro release studies demonstrated gastroresistance and a sustained drug release at pH 7.4 (97.67 ± 0.32%) in 120 h. The kinetic model used for the release of MSZ from the NP-ZN in a pH 1.2 medium was the Fickian diffusion, in a pH 6.8 medium it was the Peppas–Sahlin model with the polymeric relaxation mechanism and in a pH 7.4 medium it was the Korsmeyer–Peppas model with the Fickian release mechanism, or “Case I”. An in vitro cytotoxicity study in the CT26.WT cell line showed no basal cytotoxicity up to 500 μg/mL. The NP-ZN showed to be a promising vector for the sustained release of MSZ in the colon by oral route