RESUMO
Breast cancer is the most prevalent cancer in women worldwide. Its molecular subtypes are based on the presence/absence of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). MACL-1 and MGSO-3 are cell lines derived from primary tumor sites of patients diagnosed with luminal A subtype carcinoma (ER+/PR+/HER2-) and ductal carcinoma in situ (ER-/PR-/HER2+), respectively. However, these cell lines lost the expression of these markers over cell culturing, and both have triple-negative phenotypes (ER-/PR-/HER2-), which has the poorest prognosis. Here, we sought to study the proteome signature of MGSO-3 and MACL-1, comparing them with the epithelial cell line MCF-10A and the well-established metastatic-derived breast cancer cell line MDA-MB-231. Our results showed that proteins associated with the tricarboxylic acid cycle (TCA) and oxidative phosphorylation (OXPHOS) were upregulated in MGSO-3 and MACL-1 cells. These cell lines also showed upregulation of pro-apoptotic proteins when compared with MDA-MB-231. The molecular differences highlighted in this study may clarify the molecular basis behind cancer cells functioning and may reveal novel signatures across the breast cancer cell models.
Assuntos
Neoplasias da Mama , Carcinoma , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/metabolismo , Carcinoma/patologia , Linhagem Celular , Feminino , Humanos , Proteômica , Receptor ErbB-2/metabolismo , Receptores de Estrogênio/metabolismoRESUMO
In recent years, acid whey production has increased due to a growing demand for Greek yogurt and acid-coagulated cheeses. Acid whey is a dairy by-product for which the industry has long struggled to find a sustainable application. Bulk amounts of acid whey associated with the dairy industry have led to increasing research on ways to valorize it. Industry players are finding ways to use acid whey on-site with ultrafiltration techniques and biodigesters, to reduce transportation costs and provide energy for the facility. Academia has sought to further investigate practical uses and benefits of this by-product. Although modern research has shown many other possible applications for acid whey, no comprehensive review yet exists about its composition, utilization, and health benefits. In this review, the industrial trends, the applications and uses, and the potential health benefits associated with the consumption of acid whey are discussed. The proximal composition of acid whey is discussed in depth. In addition, the potential applications of acid whey, such as its use as a starting material in the production of fermented beverages, as growth medium for cultivation of lactic acid bacteria in replacement of commercial media, and as a substrate for the isolation of lactose and minerals, are reviewed. Finally, the potential health benefits of the major protein constituents of acid whey, bioactive phospholipids, and organic acids such as lactic acid are described. Acid whey has promising applications related to potential health benefits, ranging from antibacterial effects to cognitive development for babies to human gut health.
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Indústria de Laticínios/métodos , Promoção da Saúde , Soro do Leite/química , Animais , Queijo , Meios de Cultura/análise , Laticínios , Fermentação , Manipulação de Alimentos/métodos , Concentração de Íons de Hidrogênio , Ácido Láctico/análise , Lactobacillales/metabolismo , Lactose/análise , Proteínas do Soro do Leite/análise , IogurteRESUMO
OBJECTIVES: Allergic reactions are rare and poorly understood complications of neuromodulation device implantation. There are currently no guidelines for management of allergic reactions to these devices and their components. Here we review the published cases of allergic reactions to implanted neuromodulatory devices and leverage the experiences of other specialties that deal with similar complications to formulate recommendations for prevention and management. MATERIALS AND METHODS: A review and assessment of the literature. RESULTS: Allergic reactions to a number of implantable devices have been observed and published. In dentistry and orthopedics, metals such as nickel are the most frequent cause of allergic reactions. In interventional cardiology, where devices closely resemble neuromodulatory devices, titanium, silicone, and polyurethanes are the most common causes of allergic reactions. In neurosurgery, allergic reactions to implantable neuromodulatory devices are rare, and we summarize 13 cases published to date. Such allergic reactions generally present as local dermatitis, erythema, and pruritus, which can be difficult to distinguish from surgical site infection. In one published case, symptoms resolved with corticosteroid treatment, but all other cases required explantation. The successful reimplantation with a modified device was reported in some cases. CONCLUSIONS: Patients should be screened for a personal history of contact allergy before implantation procedures. A multidisciplinary approach to suspected cases of postoperative allergic reactions involving collaboration between neurosurgeons and other implanting physicians, dermatologists or allergists, and device manufacturers is recommended. In cases where an allergic reaction is suspected, an infectious etiology should be ruled out first. Clinical suspicion can then be supported with the use of patch testing, interpreted by an experienced dermatologist or allergist. If patch testing supports an allergic etiology, the implanting physician and the device manufacturer can work together to modify the device for safe reimplantation.
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Dermatite Alérgica de Contato , Remoção de Dispositivo , Eritema , Humanos , Testes do Emplastro , Próteses e ImplantesRESUMO
Lactic acid bacteria are the predominant group within meat products, whose metabolites such as bacteriocins and peptidoglycan hydrolases inhibit pathogenic or spoilage bacteria. Fermented meat products, as a salami, is a good source to analyze the viable microbiota, due to these products present a low risk to consumer health. The aim of this work was to identify the lactic acid bacteria with broad antibacterial activity present in salami, purify the protein responsible for this activity, achieve antagonistic spectrum and perform the biochemical characterization. Five strains from salami were selected, isolated and identified by 16S rRNA gene sequencing. The antimicrobial activity was evaluated by antagonism assay and zymography, using spoilage microorganisms commonly found in meat products. The strain that showed a broad antibacterial activity was Latilactobacillus sakei and the antibacterial activity was given by a protein with 75-kDa of molecular mass, identified by LC/MALDI-TOF/TOF. The sequence analysis showed 67% of identity with a N-acetylmuramoyl-L-alanine amidase protein with five non-identical LysM domains. The purified protein showed an optimal pH of 8.0 and heat resistance at 80 °C for 10 min. L. sakei strain displayed antibacterial activity against Gram-negative and Gram-positive spoilage microorganisms. The results of this study provide the information to use Latilactobacillus sakei as a starter culture which will provide the necessary metabolites to combat undesirable microorganisms. Additionally, the conditions and properties for the best application and use of the antibacterial protein produced by this strain. This protein may have a potential use in the food industry as a new antibacterial agent.
Assuntos
Antibacterianos/biossíntese , Antibacterianos/farmacologia , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Produtos da Carne/microbiologia , N-Acetil-Muramil-L-Alanina Amidase/biossíntese , Bactérias/efeitos dos fármacos , Bacteriocinas/farmacologia , Fermentação , Alimentos Fermentados/microbiologia , Microbiologia de Alimentos , Lactobacillus/genética , Testes de Sensibilidade Microbiana , Peso Molecular , N-Acetil-Muramil-L-Alanina Amidase/genética , N-Acetil-Muramil-L-Alanina Amidase/isolamento & purificação , RNA Ribossômico 16SRESUMO
The gene encoding N-acetylmuramoyl-L-alanine amidase in Latilactobacillus sakei isolated from a fermented meat product was cloned in two forms: its complete sequence (AmiC) and a truncated sequence without one of its anchoring LysM domains (AmiLysM4). The objective of this work was to evaluate the effect of LysM domain deletion on antibacterial activity as well the biochemical characterization of each recombinant protein. AmiC and AmiLysM4 were expressed in Escherichia coli BL21. Using a zymography method, two bands with lytic activity were observed, which were confirmed by LC-MS/MS analysis, with molecular masses of 71 kDa (AmiC) and 66 kDa (AmiLysM4). The recombinant proteins were active against Listeria innocua and Staphylococcus aureus strains. The inhibitory spectrum of AmiLysM4 was broader than AmiC as it showed inhibition of Leuconostoc mesenteroides and Weissella viridescens, both microorganisms associated with food decomposition. Optimal temperature and pH values were determined for both proteins using L-alanine-p-nitroanilide hydrochloride as a substrate for N-acetylmuramoyl-L-alanine amidase activity. Both proteins showed similar maximum activity values for pH (8) and temperature (50 °C). Furthermore, structural predictions did not show differences for the catalytic region, but differences were found for the region called 2dom-AmiLysM4, which includes 4 of the 5 LysM domains. Therefore, modification of the LysM domain offers new tools for the development of novel food biopreservatives.
Assuntos
Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Lactobacillaceae/enzimologia , N-Acetil-Muramil-L-Alanina Amidase/química , N-Acetil-Muramil-L-Alanina Amidase/farmacologia , Antibacterianos/química , Domínio Catalítico , Clonagem Molecular , Concentração de Íons de Hidrogênio , Lactobacillaceae/genética , Testes de Sensibilidade Microbiana , Modelos Moleculares , N-Acetil-Muramil-L-Alanina Amidase/genética , N-Acetil-Muramil-L-Alanina Amidase/metabolismo , Domínios Proteicos , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , TemperaturaRESUMO
Cryptic peptides (cryptides) are biologically active peptides formed after proteolysis of native precursors present in animal venoms, for example. Proteolysis is an overlooked post-translational modification that increases venom complexity. The tripeptide KPP (Lys-Pro-Pro) is a peptide encrypted in the C-terminus of Ts14-a 25-mer peptide from the venom of the Tityus serrulatus scorpion that has a positive impact on the cardiovascular system, inducing vasodilation and reducing arterial blood pressure of hypertensive rats among other beneficial effects. A previous study reported that KPP and its native peptide Ts14 act via activation of the bradykinin receptor B2 (B2R). However, the cellular events underlying the activation of B2R by KPP are unknown. To study the cell signaling triggered by the Ts14 cryptide KPP, we incubated cardiac myocytes isolated from C57BL/6 mice with KPP (10-7 mol·L-1) for 0, 5, or 30 min and explored the proteome and phosphoproteome. Our results showed that KPP regulated cardiomyocyte proteins associated with, but not limited to, apoptosis, muscle contraction, protein turnover, and the respiratory chain. We also reported that KPP led to AKT phosphorylation, activating AKT and its downstream target nitric oxide synthase. We also observed that KPP led to dephosphorylation of phospholamban (PLN) at its activation sites (S16 and T17), leading to reduced contractility of treated cardiomyocytes. Some cellular targets reported here for KPP (e.g., AKT, PLN, and ERK) have already been reported to protect the cardiac tissue from hypoxia-induced injury. Hence, this study suggests potential beneficial effects of this scorpion cryptide that needs to be further investigated, for example, as a drug lead for cardiac infarction.
Assuntos
Venenos de Escorpião , Animais , Proteínas de Ligação ao Cálcio , Camundongos , Camundongos Endogâmicos C57BL , Miócitos Cardíacos , Peptídeos/farmacologia , Proteínas Proto-Oncogênicas c-akt , Ratos , Venenos de Escorpião/farmacologia , Transdução de SinaisRESUMO
Lactic acid bacteria (LAB) are a unique subset of microorganisms that have co-evolved with humans since the beginning of agricultural practices and animal domestication and throughout our never-ending quest for food preservation, digestibility, and flavor enhancement. LAB have historically played a preponderant role in our foods. In this review, we focus on the enzymatic activities and current or potential applications of LAB in our lives. A description of each of the enzymatic systems in LAB is included. Glycosidases, which hydrolyze the most abundant food molecules and as sources of carbon, sustain the lives of organisms on Earth as well as ensure microbial innocuity by the production of lactic acid from the uniquely mammalian carbohydrate, lactose. Lipases and proteases or proteinases are of fundamental importance in food fermentations and in dairy foods for flavor development. Bacteriocins and peptidoglycan hydrolases are part of the enzymatic system of LAB that has evolved to make these bacteria fierce competitors in various microbiomes, which are highly important for the human gut. In this review, we also present an explanation on how the versatility of the genetics of LAB can adapt to the matrix where they are placed with the advantage of not having any toxicity to humans. The systematic study of LAB enzymes has allowed for some unique applications in foods and biopharmaceutical industries. Here, we summarize how different enzyme systems in LAB are classified, and thus, facilitate much-needed further studies to understand the fundamentals and translate them into applications to improve our lives.
Assuntos
Microbiologia Industrial/tendências , Lactobacillales/enzimologia , Bacteriocinas/metabolismo , Microbiologia de Alimentos , Lactobacillales/genéticaRESUMO
Lipolysis occurs during ripening of dairy products as a result of esterase or lipase activity. Lactic acid bacteria (LAB) are considered to be weakly lipolytic bacteria compared with other species. In cheeses with extended ripening periods, lipolytic LAB may have several advantages. Pediococcus acidilactici is a LAB frequently found in fermented dairy products, but no previous reports exist on their production of esterases or lipases. Our interest in the relationship of LAB and enzymatic characterization is due to the multiple reports of the benefits of LAB in the gut microbiome, particularly at the intestinal membrane. Pediococci have been characterized as probiotic and especially active in membrane interactions. The aim of this project was to purify, characterize, and identify the phosphoesterase produced by P. acidilactici originally isolated from Gouda cheese and determine its phospholipid (PL) hydrolysis profile, with a focus on increased absorption of these compounds in the human gut. Native zymograms were performed to identify a protein with lipolytic activity in the intracellular fraction of P. acidilactici. The enzyme was purified via size-exclusion HPLC, concentrated via ultrafiltration, and identified using sequence analysis in liquid chromatography (LC)-MS/MS. The purified fraction was subjected to biochemical characterization as a function of pH, temperature, ion concentration, hydrolysis of different substrates, and PL. A single protein with a molecular weight of 86 kDa and esterase activity was detected by zymography. Analysis of the LC-MS/MS results identified a putative metallophosphoesterase with a calculated molecular weight of 45.5 kDa, suggesting that this protein is active as a homodimer. The pure protein showed an optimal activity between pH 8.0 to 9.0. The optimal temperature for activity was 37°C, and the enzyme lost 15% of activity after incubation at 90°C for 1 h. This enzyme showed activity on short-chain fatty acids and exhibited high hydrolysis of phosphatidylinositol. It also hydrolyzed phosphatidylserine, phosphatidylcholine, and sphingomyelin. Phosphatidylethanolamine was hydrolyzed but with less efficiency. The characteristics and lipolytic actions exerted by this protein obtained from LAB hold promise for a potential strain of esterase or lipase that may exert human health benefits through increased digestibility and absorption of nutrients found in dairy products.
Assuntos
Queijo/microbiologia , Pediococcus acidilactici/enzimologia , Fosfoproteínas Fosfatases/isolamento & purificação , Animais , Cromatografia Líquida , Humanos , Hidrólise , Lipólise , Peso Molecular , Pediococcus acidilactici/isolamento & purificação , Fosfoproteínas Fosfatases/metabolismo , Espectrometria de Massas em TandemRESUMO
The mechanisms of bacterial adhesion to human cells involve several complex reactions and activation of genes and proteins. It has been reported that the food components in dairy matrices, such as sugar or salt, can decrease bacterial adhesion to Caco-2 cells. However, it has not been evaluated whether the bacteria grown in media supplemented with milk phospholipids (MPL) can increase or decrease the adhesion of these cells. The objective of this work was to evaluate the effects of MPL on the kinetic growth of lactic acid bacteria (LAB) and their functional characteristics as probiotics, expression of surface protein genes, and adherence to Caco-2 cells. Seven LAB strains isolated from various dairy products were characterized. Five of the tested LAB strains were able to grow in a chemically defined medium supplemented with MPL. Lactobacillus reuteri OSU-PECh-48 showed the highest growth rate and the greatest optical density. All of the strains tested showed tolerance to acidic conditions at pH 3.0 and to bile salts at 0.5 and 1% concentrations. Auto-aggregation and cell surface hydrophobicity ability were evaluated, with nonsignificant differences between the strains grown in MPL and without MPL. Gene expression of 6 surface proteins was evaluated in the presence or absence of MPL. Pediococcus acidilactici OSU-PECh-L and OSU-PECh-48 were the strains with highest relative expression of 5 of the 6 genes evaluated. Lactobacillus paracasei OSU-PECh-BA was the strain with the lowest level of expression of surface protein genes. Most of the bacteria tested had increased adhesion to Caco-2 cells after growth in MPL. The bacteria with the highest degrees of adhesion observed were Lactobacillus paracasei OSU-PECh-3B, Pediococcus acidilactici OSU-PECh-L, and Lactobacillus reuteri OSU-PECh-48. The genes Cnb and EF-Tu increased in expression in the presence of MPL in most of the LAB tested. The results obtained in this work demonstrate the high potential of these LAB strains for use as starters or beneficial cultures in fermentation of not only dairy products but also other food fermentation processes, with promising ability to increase residence time in the gut, modify the microbiome, and improve human health.
Assuntos
Aderência Bacteriana , Meios de Cultura/metabolismo , Lactobacillales/fisiologia , Leite/microbiologia , Fosfolipídeos/metabolismo , Probióticos/metabolismo , Animais , Células CACO-2 , Fermentação , Humanos , Lactobacillales/crescimento & desenvolvimento , Lacticaseibacillus paracasei/crescimento & desenvolvimento , Lacticaseibacillus paracasei/fisiologia , MicrobiotaRESUMO
Hydroxycinnamic acid (HCA) decarboxylation by lactic acid bacteria (LAB) results in the production of 4-vinylplenols with great impact on the sensorial characteristics of foods. The determination of LAB decarboxylating capabilities is key for optimal strain selection for food production. The activity of LAB strains from the Ohio State University-Parker Endowed Chair (OSU-PECh) collection potentially capable of synthesizing phenolic acid decarboxylase was evaluated after incubation with HCAs for 36 h at 32 °C. A high-throughput method for monitoring HCAs decarboxylation was developed based on hypsochromic shifts at pH 1.0. Out of 22 strains evaluated, only Enterococcus mundtii, Lactobacillus plantarum and Pediococcus pentosaceus were capable of decarboxylating all p-coumaric, caffeic and ferulic acids. Other strains only decarboxylated p-coumaric and caffeic acid (6), only p-coumaric acid (2) or only caffeic acid (1), while 10 strains did not decarboxylate any HCA. p-Coumaric acid had the highest conversion efficiency, followed by caffeic acid and lastly ferulic acid. Results were confirmed by HPLC-DAD-ESI-MS analyses, showing the conversion of HCAs into their 4-vinylphenol derivatives. This work can help improve the sensory characteristics of HCA-rich foods where fermentation with LAB was used during processing.
Assuntos
Ácidos Cumáricos/metabolismo , Microbiologia de Alimentos , Lactobacillales/metabolismo , Descarboxilação , Espectrofotometria UltravioletaRESUMO
Ischemic heart disease is the leading cause of deaths worldwide. Thus, understanding the molecular mechanisms underlying disease progression is needed. Due to heart importance and lack of studies evaluating different sample preparation methods for heart proteomics, we compared three well-established protocols in shotgun proteomics using dimethyl label quantitation to allow relative quantitation. The tested methods for the analysis of left ventricle (LV) tissue were: i) in-solution digestion (ISD); ii) on-pellet digestion (OPD); and iii) on-filter digestion (OFD). Protein extraction was done using SDS-containing buffer for OPD and OFD while this step was under urea-containing buffer for ISD. We used an optimized one-step reaction for reduction of disulfide bonds and alkylation of thiol groups in ISD and OPD. Using the same amount of tissue, we observed that OFD and ISD extracted significantly higher amount of protein than OPD. ISD outperformed OFD and OPD in the number of proteins identified. We did not observe significant bias related to physicochemical features of the identified proteins when comparing the three protocols. ISD was more efficient to identify low abundant proteins and yielded more proteins per protocol duration. Thus, we concluded that the optimized ISD suited better for heart proteomics than OFD and OPD.
Assuntos
Ventrículos do Coração/metabolismo , Proteínas/análise , Proteômica/métodos , Animais , Masculino , Ratos Wistar , Manejo de EspécimesRESUMO
Regular consumption of fermented dairy products helps maintain a healthy microbiota and prevent gut dysbiosis-linked diseases. The lactic acid bacteria (LAB) present in food enhance the digestibility of proteins, moderate the release of fatty acids, and support human health through inhabiting the gastrointestinal tract. These desirable properties of LAB are attributed, in part, to their metabolic processes involving enzymes such as lipases, proteases, and antibacterial proteins. The LAB strains presenting higher enzymatic activities may offer improved functionality for applications in foods. The first aim of this work was to isolate and identify LAB from diverse dairy products and select those with enhanced enzymatic activities. Secondly, this work aimed to investigate the subcellular organization and identity of these enzymes after semi-purification. Out of the total 137 LAB strains isolated and screened, 50.3% and 61.3% of the strains exhibited lipolytic and proteolytic activities, respectively. Seven strains displaying high enzymatic activities were selected and further characterized for the cellular organization of their lipases, proteases, and antibacterial proteins. The lipolytic and proteolytic activities were exhibited predominantly in the extracellular fraction; whereas, the antibacterial activities were found in various cellular fractions and were capable of inhibiting common undesirable microorganisms in foods. In total, two lipases, seven proteases, and three antibacterial proteins were identified by LC-MS/MS. Characterization of LAB strains with high enzymatic activity has potential biotechnological significance in fermentative processes and in human health as they may improve the physicochemical characteristics of foods and displace strains with weaker enzymatic activities in the human gut microbiota.
Assuntos
Antibacterianos/farmacologia , Laticínios/microbiologia , Lactobacillales/enzimologia , Lactobacillales/isolamento & purificação , Lipólise , Proteólise , Antibacterianos/isolamento & purificação , Produtos Fermentados do Leite/microbiologia , Escherichia coli/efeitos dos fármacos , Fermentação , Microbiologia de Alimentos , Lipase/isolamento & purificação , Lipase/metabolismo , Peptídeo Hidrolases/isolamento & purificação , Peptídeo Hidrolases/metabolismo , Staphylococcus aureus/efeitos dos fármacosRESUMO
While the cooking-culture relationship is a biocultural phenomenon that gives meaning to gastronomic practices through signs and symbols shared within a community, in the 20th century, its transmission has been influenced by important processes of change, such as urban expansion and environmental pollution, which put the continuity of immaterial cultural patrimony at risk. These changes have altered the supply, conservation, and preparation of food and utensils, which are elements that demarcate the society-nature relationship and gender roles, among other things. The objective of this article is to demonstrate the role of the cultural transmission of food habits in identity formation and social cohesion, based upon an ethnographic case study in the Ancestral Community La Playa Renaciente, in Cali (Colombia). This study presents the cultural transmission of food habits, culinary practices, and their relationship with the inhabited environment. Likewise, this article presents a temporal-spatial contextualization of the locality and some oral testimonies that reveal how dietary practices constitute an important element in socialization, social cohesion, and the transmission of knowledge from generation to generation.
Assuntos
Evolução Cultural , Comportamento Alimentar/etnologia , Indígenas Sul-Americanos/psicologia , Ajustamento Social , Identificação Social , Colômbia/etnologia , Culinária , Humanos , Indígenas Sul-Americanos/etnologia , População RuralRESUMO
OBJECTIVE: To provide a brief overview of the clinical presentation, common offending agents, management, prognosis, and mortality of 6 selected high-risk drug rashes, namely, Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome, multiple drug hypersensitivity (MDH) syndrome, acute generalized exanthematous pustulosis (AGEP), and drug-induced bullous pemphigoid (DIBP). DATA SOURCES: A review of the published literature was performed with PubMed and supplemented with our clinical experience. STUDY SELECTIONS: The most recent clinically relevant studies and older seminal works were selected. RESULTS: Most of the published data on these uncommon rashes were based on small observational series or case reports. SJS and TEN have specific genotypes association with certain drugs, have high morbidity and mortality, and require aggressive management by a team of multiple specialists. DRESS syndrome is a severe, prolonged multiorgan reaction, yet it has a better prognosis than TEN. MDH is a syndrome of repeated reactions to unrelated drugs that often imposes diagnostic and management difficulties. AGEP consists of generalized sterile small pustules, usually mistaken for infection with subsequent inappropriate treatment. Bullous pemphigoid presents with tense pruritic bullae and characteristic linear basement membrane deposition of IgG and C3. DIBP has much better prognosis than the autoimmune variety. CONCLUSION: In such high-risk drug rashes, early recognition, immediate withdrawal of the suspected drug(s), prompt individualized management, and monitoring of vital organs function are mandatory for reducing morbidity and mortality. The lack of reliable tests for identification of the causative agent imposes difficulty, particularly in patients receiving multiple medications.
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Toxidermias/etiologia , Toxidermias/mortalidade , Toxidermias/diagnóstico , Toxidermias/terapia , Humanos , Fatores de RiscoRESUMO
Glucokinases (Glks) are enzymes widely distributed in all three domains of life. They are located at the beginning of the glycolytic pathway and are responsible for the glucose phosphorylation from various phosphate group donors such as ATP, ADP and polyphosphate. So far, there are eight crystallized Glks, and at least one belongs to each of the three reported Glk families. Structural studies have elucidated the mechanism for Glk action and multimerization. Cloning, overexpression and biochemical characterization have demonstrated the wide diversity of these enzymes. As reported for various microorganisms, in addition to their catalytic activity, some Glks, possessing ROK (Repressor Orf Kinases) motifs, also display a regulatory role. This function has been associated to the mechanisms of carbon catabolite regulation, morphological differentiation and antibiotic production. The present review covers the classification, detailed tertiary structure, mechanism of action, biochemical characterization and some regulatory aspects of bacterial Glks.
Assuntos
Bactérias/enzimologia , Glucoquinase/metabolismo , Bactérias/química , Bactérias/genética , Bactérias/metabolismo , Clonagem Molecular/métodos , Glucoquinase/química , Glucoquinase/genética , Conformação ProteicaRESUMO
The present work aims at characterizing T. harzianum secretome when the fungus is grown in synthetic medium supplemented with one of the four substrates: glucose, cellulose, xylan, and sugarcane bagasse (SB). The characterization was done by enzymatic assays and proteomic analysis using 2-DE/MALDI-TOF and gel-free shotgun LC-MS/MS. The results showed that SB induced the highest cellulolytic and xylanolytic activities when compared with the other substrates, while remarkable differences in terms of number and distribution of protein spots in 2-DE gels were also observed among the samples. Additionally, treatment of the secretomes with PNGase F revealed that most spot trails in 2-DE gels corresponded to N-glycosylated proteoforms. The LC-MS/MS analysis of the samples identified 626 different protein groups, including carbohydrate-active enzymes and accessory, noncatalytic, and cell-wall-associated proteins. Although the SB-induced secretome displayed the highest cellulolytic and xylanolytic activities, it did not correspond to a higher proteome complexity because CM-cellulose-induced secretome was significantly more diverse. Among the identified proteins, 73% were exclusive to one condition, while only 5% were present in all samples. Therefore, this study disclosed the variation of T. harzianum secretome in response to different substrates and revealed the diversity of the fungus enzymatic toolbox.
Assuntos
Biomassa , Proteínas Fúngicas/análise , Proteoma/análise , Trichoderma/enzimologia , Trichoderma/metabolismo , Celulase , Celulose , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Mapeamento de Peptídeos , Proteoma/química , Proteoma/metabolismo , Proteômica/métodos , Trichoderma/química , Trichoderma/fisiologiaRESUMO
Tranexamic acid (TXA), a fibrinolytic agent, effectively inhibits plasminogen activation, thereby reducing fibrinolysis and hemorrhage. This study focused on its application in trauma patients undergoing emergency surgery, a critical area due to trauma's significant role in mortality. Our investigation involved a meticulous screening of randomized controlled trials from databases including Scopus, PubMed, Web of Science, and Cochrane. The findings indicate that TXA intervention is promising in enhancing outcomes for trauma patients. However, the drug's effectiveness may vary based on the specific nature of the medical condition. In summary, robust evidence suggests that TXA can diminish blood loss, lower transfusion rates, reduce complications, and improve hemoglobin and hematocrit levels in surgical patients. Consequently, TXA should be considered a crucial medication, readily available to mitigate morbidity and mortality in surgical settings. Future research should explore factors influencing TXA's effectiveness in traumatic brain injury cases and across a broad spectrum of surgical scenarios in diverse patient populations. This would further guide clinicians in refining and optimizing the use of TXA.
RESUMO
Hypertrophic cardiomyopathy (HCM) is a hereditary cardiac condition characterized by unexplained left ventricular hypertrophy without a hemodynamic cause. This condition is prevalent in the United States, resulting in various clinical manifestations, including diastolic dysfunction, left ventricular outflow obstruction, cardiac ischemia, and atrial fibrillation. HCM is associated with several genetic mutations, with sarcomeric mutations being the most common and contributing to a more complex disease course. Early diagnosis of HCM is essential for effective management, as late diagnosis often requires invasive treatments and creates a substantial financial burden. Disparities in HCM diagnosis and treatment exist between high-income and low-income countries. High-income countries have more resources to investigate and implement advanced diagnostic and treatment modalities. In contrast, low-income countries face challenges in accessing diagnostic equipment, trained personnel, and affordable medications, leading to a lower quality of life and life expectancy for affected individuals. Diagnostic tools for HCM include imaging studies such as 2D echocardiography, cardiovascular magnetic resonance (CMR), and electrocardiograms (ECGs). CMR is considered the gold standard but remains inaccessible to a significant portion of the world's population, especially in low-income countries. Genetics plays a crucial role in HCM, with numerous mutations identified in various genes. Genetic counseling is essential but often limited in low-income countries due to resource constraints. Disparities in healthcare access and adherence to treatment recommendations exist between high-income and low-income countries, leading to differences in patient outcomes. Addressing these disparities is essential to improve the overall management of HCM on a global scale. In conclusion, this review highlights the complex nature of HCM, emphasizing the importance of early diagnosis, genetic counseling, and access to appropriate diagnostic and therapeutic interventions. Addressing healthcare disparities is crucial to ensure that all individuals with HCM receive timely and effective care, regardless of their geographic location or socioeconomic status.
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Intrapericardial teratoma is a rare tumor composed of tissue from the three germ cell layers with a rapid growth that may cause severe hemodynamic complications due to compressive effects. We present two clinical cases: the first case had severe fetal heart failure with fatal outcome, and the second underwent surgical treatment during the immediate postnatal period with a favorable evolution. Although teratomas are histologically benign tumors, rapid growth can cause serious hemodynamic complications. The importance of prenatal diagnosis is to allow appropriate monitoring of tumor growth and establish a prompt therapeutic plan. Opportune surgical treatment can prevent death and improve the prognosis of these patients.
Assuntos
Neoplasias Cardíacas , Teratoma , Gravidez , Feminino , Humanos , Pericárdio/cirurgia , Pericárdio/diagnóstico por imagem , Diagnóstico Pré-Natal , Teratoma/diagnóstico por imagem , Teratoma/cirurgia , Prognóstico , Neoplasias Cardíacas/diagnóstico por imagem , Neoplasias Cardíacas/cirurgia , Ultrassonografia Pré-NatalRESUMO
Aim: To develop, characterize and evaluate an oil/water nanoemulsion with squalene (CTVad1) to be approved as an adjuvant for the SpiN COVID-19 vaccine clinical trials. Materials & methods: Critical process parameters (CPPs) of CTVad1 were standardized to meet the critical quality attributes (CQAs) of an adjuvant for human use. CTVad1 and the SpiN-CTVad1 vaccine were submitted to physicochemical, stability, in vitro and in vivo studies. Results & conclusion: All CQAs were met in the CTVad1 production process. SpiN- CTVad1 met CQAs and induced high levels of antibodies and specific cellular responses in in vivo studies. These results represented a critical step in the process developed to meet regulatory requirements for the SpiN COVID-19 vaccine clinical trial.