Innovations and developments in single cell protein: Bibliometric review and patents analysis

BackgroundGlobal demand for food products derived from alternative proteins and produced through sustainable technological routes is increasing. Evaluation of research progress, main trends and developments in the field are valuable to identify evolutionary nuances.MethodsIn this study, a bibliometric analysis and search of patents on alternative proteins from fermentation processes was carried out using the Web of Science and Derwent World Patents Index™ databases, using the keywords and Boolean operators “fermentation” AND “single cell protein” OR “single-cell protein.” The dataset was processed and graphics generated using the bibliometric software VOSviewer and OriginPro 8.1.ResultsThe analysis performed recovered a total of 360 articles, of which 271 were research articles, 49 literature review articles and 40 publications distributed in different categories, such as reprint, proceedings paper, meeting abstract among others. In addition, 397 patents related to the field were identified, with China being the country with the largest number of publications and patents deposits. While this topic is largely interdisciplinary, the majority of work is in the area of Biotechnology Applied Microbiology, which boasts the largest number of publications. The area with the most patent filings is the food sector, with particular emphasis on the fields of biochemistry, beverages, microbiology, enzymology and genetic engineering. Among these patents, 110 are active, with industries or companies being the largest depositors. Keyword analysis revealed that the area of study involving single cell protein has included investigation into types of microorganisms, fermentation, and substrates (showing a strong trend in the use of agro-industrial by-products) as well as optimization of production processes.ConclusionThis bibliometric analysis provided important information, challenges, and trends on this relevant subject

Xylopine Induces Oxidative Stress and Causes G 2

Xylopine is an aporphine alkaloid that has cytotoxic activity to cancer cells. In this study, the underlying mechanism of xylopine cytotoxicity was assessed in human colon carcinoma HCT116 cells. Xylopine displayed potent cytotoxicity in different cancer cell lines in monolayer cultures and in a 3D model of cancer multicellular spheroids formed from HCT116 cells. Typical morphology of apoptosis, cell cycle arrest in the G2/M phase, increased internucleosomal DNA fragmentation, loss of the mitochondrial transmembrane potential, and increased phosphatidylserine externalization and caspase-3 activation were observed in xylopine-treated HCT116 cells. Moreover, pretreatment with a caspase-3 inhibitor (Z-DEVD-FMK), but not with a p53 inhibitor (cyclic pifithrin-α), reduced xylopine-induced apoptosis, indicating induction of caspase-mediated apoptosis by the p53-independent pathway. Treatment with xylopine also caused an increase in the production of reactive oxygen/nitrogen species (ROS/RNS), including hydrogen peroxide and nitric oxide, but not superoxide anion, and reduced glutathione levels were decreased in xylopine-treated HCT116 cells. Application of the antioxidant N-acetylcysteine reduced the ROS levels and xylopine-induced apoptosis, indicating activation of ROS-mediated apoptosis pathway. In conclusion, xylopine has potent cytotoxicity to different cancer cell lines and is able to induce oxidative stress and G2/M phase arrest, triggering caspase-mediated apoptosis by the p53-independent pathway in HCT116 cells

A ruthenium-based 5-fluorouracil complex with enhanced cytotoxicity and apoptosis induction action in HCT116 cells.

Combination of multifunctionalities into one compound is a rational strategy in medicinal chemical design, and have often been used with metallodrug-based compounds. In the present study, we synthesized a novel ruthenium-based 5-fluorouracil complex [Ru(5-FU)(PPh3)2(bipy)]PF6 (PPh3?=?triphenylphosphine; and bipy?=?2,2?-bipyridine) with enhanced cytotoxicity in different cancer cells, and assessed its apoptosis induction action in human colon carcinoma HCT116 cells. The complex was characterized by infrared, cyclic voltammetry, molar conductance measurements, elemental analysis, NMR experiments and X-ray crystallographic analysis. In both 2D and 3D cell culture models, the complex presented cytotoxicity to cancer cells more potent than 5-FU. A typical morphology of apoptotic cell death, increased internucleosomal DNA fragmentation, without cell membrane permeability, loss of the mitochondrial transmembrane potential, increased phosphatidylserine externalization and caspase-3 activation were observed in complex-treated HCT116 cells. Moreover, the pre-treatment with Z-DEVD-FMK, a caspase-3 inhibitor, reduced the apoptosis induced by the complex, indicating cell death by apoptosis through caspase-dependent and mitochondrial intrinsic pathways. The complex failed to induce reactive oxygen species production and DNA intercalation. In conclusion, the novel complex displays enhanced cytotoxicity to different cancer cells, and is able to induce caspase-mediated apoptosis in HCT116 cells

Avaliação da atividade imunomoduladora de extratos de peles e glândulas parotóides de anuros do semi-árido brasileiro

The skin of amphibians has been characterized by the presence of several chemical products with diverse biological activities. These compounds participate as defense mechanisms against pathogens and harmful substances to which amphibians are exposed in their natural habitats. This work aimed to investigate the pharmacological activity of skin and gland extracts prepared from species of anures native or endemic from Brazilian semi-arid region with emphasis on immunomodulatory activity. Aqueous extracts were prepared by homogenization of skins of seven anure species (Hypsiboas crepitans, Hyspsiboas albopunctatus, Bokermannohyla oxente, Leptodactylus ocellatus, Chaunus rubescens, Ceratophrys joazeirensis e Chaunus jimi) and of glands of two species (Chaunus rubescens and Chaunus jimi). To evaluate the inhibition of nitric oxide (NO) production by the extracts, we used J774 cells stimulated with IFN-g and LPS and measured the nitrite concentration using the Griess method 24 hour later. For lymphoproliferation assays, BALB/c mice spleen cells were stimulated in vitro with concanavalin A in the presence or absence of extracts being the proliferative response determined by 3H-thymidine uptake quantification. Of the nine extracts analyzed, five had inhibitory activity superior to 50% in lymphoproliferation assay (skin extract of C. rubescens, B. oxente, Chaunus. jimi, L. ocellatus and gland extract of C. rubescens) and only one extract inhibited NO production (Chaunus rubescens). The results demonstrate that anure species from the Brazilian semi-arid are a potential source of molecules with immunomodulatory activity. Studies aiming to isolate and characterize the active molecule in the skin of C. rubescens are being carried out.A pele dos anfíbios possui uma variedade de produtos químicos com diversas atividades biológicas. Esses compostos participam dos mecanismos de defesa contra patógenos e substâncias estranhas aos quais os anfíbios são expostos em seu habitat. Esse trabalho teve o objetivo de investigar a atividade farmacológica de extratos de peles e glândulas de anuros do semi-árido brasileiro visando à identificação de substâncias com atividade imunomoduladora. Extratos aquosos foram preparados a partir de homogeneização das peles de sete espécies de anuros Hypsiboas crepitans, Hypsiboas albopunctatus, Bokermannohyla oxente, Leptodactylus ocellatus, Chaunus rubescens, Ceratophrys joazeirensis e Chaunus jimi) e das glândulas de duas espécies de anuros (Chaunus rubescens e Chaunus jimi). Para avaliar a inibição da produção de óxido nítrico (NO) pelos extratos, foram utilizadas células da linhagem J774 estimuladas com IFN-g e LPS, medindo-se a concentração de nitrito após 24 horas pelo método de Griess. Nos ensaios de linfoproliferação, esplenócitos de camundongos BALB/c foram estimulados com concanavalina A na presença ou ausência dos extratos, determinando-se a proliferação por meio da avaliação da incorporação de 3H-timidina. Dos nove extratos analisados, cinco apresentaram atividade inibitória superior a 50% no ensaio de linfoproliferação (extrato de pele de C. rubescens, B. oxente, C. jimi, L. ocellatus e de glândula de C. rubescens) e apenas um extrato inibiu a produção de NO (C. rubescens). Os resultados demonstram que os anuros do semi-árido são fontes potenciais de moléculas imunossupressoras. Outros estudos estão sendo realizados com o objetivo de isolar e caracterizar as moléculas ativas na pele de C. rubescens

Anticonvulsant activity of bone marrow cells in electroconvulsive seizures in mice

Background: Bone marrow is an accessible source of progenitor cells, which have been investigated as treatment for neurological diseases in a number of clinical trials. Here we evaluated the potential benefit of bone marrow cells in protecting against convulsive seizures induced by maximum electroconvulsive shock (MES), a widely used model for screening of anti-epileptic drugs. Behavioral and inflammatory responses were measured after MES induction in order to verify the effects promoted by transplantation of bone marrow cells. To assess the anticonvulsant effects of bone marrow cell transplantation, we measured the frequency and duration of tonic seizure, the mortality rate, the microglial expression and the blood levels of cytokine IL-1, IL-6, IL-10 and TNF-alpha after MES induction. We hypothesized that these behavioral and inflammatory responses to a strong stimulus such as a convulsive seizure could be modified by the transplantation of bone marrow cells.Results: Bone marrow transplanted cells altered the convulsive threshold and showed anticonvulsant effect by protecting from tonic seizures. Bone marrow cells modified the microglial expression in the analyzed brain areas, increased the IL-10 and attenuate IL-6 levels.Conclusions: Bone marrow cells exert protective effects by blocking the course of electroconvulsive seizures. Additionally, electroconvulsive seizures induced acute inflammatory responses by altering the pattern of microglia expression, as well as in IL-6 and IL-10 levels. Our findings also indicated that the anticonvulsant effects of these cells can be tested with the MES model following the same paradigm used for drug testing in pharmacological screening. Studies on the inflammatory reaction in response to acute seizures in the presence of transplanted bone marrow cells might open a wide range of discussions on the mechanisms relevant to the pathophysiology of epilepsies.Associacao Beneficente de Coleta de Sangue da Fundacao de Apoio a Pesquisa- UNIFESP (FAP-Colsan)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, UNIFESP, Dept Fisiol, Lab Neurofisiol, BR-04023066 São Paulo, BrazilUniversidade Federal de São Paulo, Disciplina Imunol, Dept Microimunoparasitol, São Paulo, BrazilFundacao Oswaldo Cruz, Ctr Pesquisas Goncalo Moniz, Salvador, BA, BrazilHosp Sao Rafael, Ctr Biotecnol & Terapia Celular, Salvador, BA, BrazilUniversidade Federal de São Paulo, UNIFESP, Dept Fisiol, Lab Neurofisiol, BR-04023066 São Paulo, BrazilUniversidade Federal de São Paulo, Disciplina Imunol, Dept Microimunoparasitol, São Paulo, BrazilWeb of Scienc

A randomized clinical trial to evaluate the efficacy of L-carnitine L-tartrate to modulate the effects of SARS-CoV-2 infection

IntroductionL-carnitine (LC) has been associated with inflammatory mediator reduction and with downregulating the angiotensin-converting enzyme-2 (ACE2) receptor, which is the target of SARS-CoV-2 attachment.MethodsThis pilot phase 2 randomized, double-blind placebo-controlled trial contained two cohorts. Cohort 1 comprised 101 individuals with negative RT-PCR SARS-CoV-2 test results who cohabitated with an individual diagnosed with SARS-CoV-2 infection. Cohort 2 comprised 122 individuals with positive SARS-CoV-2 RT-PCR test results who were asymptomatic or had mild COVID-19 pneumonia symptoms. Participants in each cohort were randomized 1:1 to receive either 2 g elemental oral LC supplementation or placebo daily for 21 days. Primary endpoints included adverse events, SARS-CoV-2 infection incidence in Cohort 1, and disease progressions in Cohort 2. Secondary endpoints included between-group laboratory profile comparisons and Cohort 2 ACE1/ACE2 plasma levels. Disease progression was compared between the Cohort 2 groups using chest computed tomography.ResultsIn Cohort 1, two SARS-CoV-2 infections occurred in each group. The common adverse events included headache, dyspnea, and tiredness. In Cohort 2, platelet counts were elevated, and fibrinogen levels reduced in the LC group compared with those of the placebo group.ConclusionOur study showed that LC was well-tolerated and suggests it modulates coagulation pathways. Furthermore, chest computed tomography images of the Cohort 2 LC group showed significant lung lesion improvement, suggesting that LC may slow COVID-19 progression

Nucleobase derivatives as building blocks to borm Ru(II)-based complexes with high cytotoxicity.

Two new Ru(II)-based complexes containing 2-thiouracil derivatives, known as 2-thiouracil (2TU) and 6-methyl-2-thiouracil (6m2TU), were synthesized using cis,trans-[RuCl2(PPh3)2(bipy)] as a precursor. The obtained compounds with a general formula trans-[Ru(2TU)(PPh3)2(bipy)]PF6 (1) and trans-[Ru(6m2TU)(PPh3)2(bipy)]PF6 (2) were characterized by analytical techniques such as NMR, UV?vis, and IR spectroscopies, elementary analysis, mass spectrometry, and single-crystal X-ray diffraction. Moreover, the investigation of the complexes?DNA interaction were carried out using spectrophotometric titrations and showed that the complexes present a weak interaction with this biomolecule. The compounds were evaluated against HL-60, K-562, HepG2, and B16-F10 cancer cells and against noncancer cells (PBMCs). The results of the biological assay revealed that complex 2 is more promising than complex 1. Finally, the present study suggests that complexes 1 and 2 causes cell death by apoptosis, significantly increasing the percentage of apoptotic HL-60 cells, in which the compounds altered the cell cycle, reducing the cells in G1/G0, G2/M, and S phases

Arming Mesenchymal Stromal/Stem Cells Against Cancer: Has the Time Come?

Since mesenchymal stromal/stem cells (MSCs) were discovered, researchers have been drawn to study their peculiar biological features, including their immune privileged status and their capacity to selectively migrate into inflammatory areas, including tumors. These properties make MSCs promising cellular vehicles for the delivery of therapeutic molecules in the clinical setting. In recent decades, the engineering of MSCs into biological vehicles carrying anticancer compounds has been achieved in different ways, including the loadingof MSCs with chemotherapeutics or drug functionalized nanoparticles (NPs), genetic modifications to force the production of anticancer proteins, and the use of oncolytic viruses. Recently, it has been demonstrated that wild-type and engineered MSCs can release extracellular vesicles (EVs) that contain therapeutic agents. Despite the enthusiasm for MSCs as cyto-pharmaceutical agents, many challenges, including controlling the fate of MSCs after administration, must still be considered. Preclinical results demonstrated that MSCs accumulate in lung, liver, and spleen, which could prevent their engraftment into tumor sites. For this reason, physical, physiological, and biological methods have been implemented to increase MSC concentration in the target tumors. Currently, there are more than 900 registered clinical trials using MSCs. Only a small fraction of these are investigating MSC-based therapies for cancer, but the number of these clinical trials is expected to increase as technology and our understanding of MSCs improve. This review will summarize MSC-based antitumor therapies to generate an increasing awareness of their potential and limits to accelerate their clinical translation

Chemical composition, larvicidal and cytotoxic activity of Annona salzmannii (Annonaceae) seed oil

The seed oil of Annona salzmannii A. DC. was analyzed by GC-MS and 1 H qNMR, revealing a mixture of unsaturated (80.5%) and saturated (18.7%) fatty acids. Linoleic (45.3%) and oleic (33.5%) acid were the major unsaturated fatty acids identified, while palmitic acid (14.3%) was the major saturated fatty acid. The larvicidal effects of A. salzmannii seed oil were evaluated against third-instar larvae of Aedes aegypti (Linn.). The oil exhibited moderate larvicidal activity, with aLC50 of 569.77 ppm (95% CI = 408.11 to 825.88 ppm). However, when the cytotoxic effects of the oil were evaluated, no expressive antiproliferative effects were observed in tumor cell lines B16-F10 (mouse melanoma), HepG2 (human hepatocellular carcinoma), K562 (human chronic myelocytic leukemia), HL-60 (human promyelocytic leukemia), and non-tumor cell line PBMC (peripheral blood mononuclear cells), with IC50 values > 50 μg·mL-1. This is the first study to evaluate the chemical composition, larvicidal and cytotoxic activity of A. salzmannii seed oil