RESUMO
Extracellular matrix (ECM) is playing a critical role which is component of mammalian tissue that provide structural support to cells. In addition, ECM act as a local depot for growth factors that control cell phenotype and differentiation. In this regard, scaffold that mimicking the ECM structure is important to growth or wound healing process. Gelatin is natural polymer and derived from collagen which is a major component of ECM. Using gelatin as an ECM mimicking structure has advantage of providing three-dimensional growth or supporting to regulate the cell behavior, proliferation, migration, cell survival, and differentiation. In this study, we developed enzyme-mediated crosslinking gelatin-based hydrogels with robust mechanical property to mimicking ECM and effectively attach to the surrounding tissue with high adhesive property. The effect of different concentration of graphene oxide (GO) on the physico-chemical properties of gelatin hydrogels were investigated, particularly tissue adhesion strength. In vitro proteolytic degradation behavior and human dermal fibroblast proliferation study confirmed the hydrogels were biodegradable and promote cell proliferation. Overall, we suggest that GO incorporated gelatin hydrogels with additional interfacial interactions, showing a promising potential as an injectable tissue adhesive.
Assuntos
Gelatina/farmacologia , Grafite/farmacologia , Hidrogéis/farmacologia , Regeneração/fisiologia , Adesivos Teciduais/farmacologia , Animais , Humanos , Espectroscopia Fotoeletrônica , Proteólise , Regeneração/efeitos dos fármacos , Sus scrofa , Resistência à TraçãoRESUMO
Engineered polymeric hydrogels have been extensively utilized in tissue engineering and regenerative medicine because of their biocompatibility, tunable properties, and structural similarity in their native extracellular microenvironment. The native extracellular matrix (ECM) has been implicated as a crucial factor in the regulation of cellular behaviors and their fate. The emerging trend in the design of hydrogels involves the development of advanced materials to precisely recapitulate the native ECM or to stimulate the surrounding tissues via physical, chemical, or biological stimuli. The ECM presents various parameters such as ECM components, soluble factors, cell-to-cell and cell-to-matrix interactions, physical forces, and physicochemical environments. Among these environmental factors, oxygen is considered as an essential signaling molecule. In particular, abnormal oxygen tension such as a lack of oxygen (defined as hypoxia) and an excess supply of oxygen (defined as hyperoxia) plays a pivotal role during early vascular development, tissue regeneration and repair, and tumor progression and metastasis. In this chapter, we discuss how engineered polymeric hydrogels serve as either an artificial extracellular microenvironment to create engineered tissues or as an acellular matrix to stimulate the native tissues for a wide range of biomedical applications including tissue engineering and regenerative medicine, wound healing, and engineered disease models. Specifically, we focus on emerging technologies to create advanced polymeric hydrogel materials that accurately mimic or stimulate the native ECM.
Assuntos
Hidrogéis , Oxigênio , Medicina Regenerativa , Engenharia Tecidual , Matriz Extracelular/química , Hidrogéis/química , Oxigênio/metabolismo , Medicina Regenerativa/métodos , Engenharia Tecidual/métodosRESUMO
Hypoxia is a critical factor in the progression and metastasis of many cancers, including soft tissue sarcomas. Frequently, oxygen (O2) gradients develop in tumors as they grow beyond their vascular supply, leading to heterogeneous areas of O2 depletion. Here, we report the impact of hypoxic O2 gradients on sarcoma cell invasion and migration. O2 gradient measurements showed that large sarcoma mouse tumors (>300 mm(3)) contain a severely hypoxic core [≤0.1% partial pressure of O2 (pO2)] whereas smaller tumors possessed hypoxic gradients throughout the tumor mass (0.1-6% pO2). To analyze tumor invasion, we used O2-controllable hydrogels to recreate the physiopathological O2 levels in vitro. Small tumor grafts encapsulated in the hydrogels revealed increased invasion that was both faster and extended over a longer distance in the hypoxic hydrogels compared with nonhypoxic hydrogels. To model the effect of the O2 gradient accurately, we examined individual sarcoma cells embedded in the O2-controllable hydrogel. We observed that hypoxic gradients guide sarcoma cell motility and matrix remodeling through hypoxia-inducible factor-1α (HIF-1α) activation. We further found that in the hypoxic gradient, individual cells migrate more quickly, across longer distances, and in the direction of increasing O2 tension. Treatment with minoxidil, an inhibitor of hypoxia-induced sarcoma metastasis, abrogated cell migration and matrix remodeling in the hypoxic gradient. Overall, we show that O2 acts as a 3D physicotactic agent during sarcoma tumor invasion and propose the O2-controllable hydrogels as a predictive system to study early stages of the metastatic process and therapeutic targets.
Assuntos
Oxigênio/metabolismo , Sarcoma/patologia , Animais , Hipóxia Celular , Movimento Celular , Hidrogéis , Camundongos , Minoxidil/farmacologia , Invasividade NeoplásicaRESUMO
Recent research has demonstrated that tumor microenvironments play pivotal roles in tumor development and metastasis through various physical, chemical, and biological factors, including extracellular matrix (ECM) composition, matrix remodeling, oxygen tension, pH, cytokines, and matrix stiffness. An emerging trend in cancer research involves the creation of engineered three-dimensional tumor models using bioinspired hydrogels that accurately recapitulate the native tumor microenvironment. With recent advances in materials engineering, many researchers are developing engineered tumor models, which are promising platforms for the study of cancer biology and for screening of therapeutic agents for better clinical outcomes. In this review, we discuss the development and use of polymeric hydrogel materials to engineer native tumor ECMs for cancer research, focusing on emerging technologies in cancer engineering that aim to accelerate clinical outcomes.
Assuntos
Materiais Biomiméticos/química , Matriz Extracelular/química , Hidrogéis/química , Neoplasias/química , Engenharia Tecidual/métodos , Microambiente Tumoral , Animais , HumanosRESUMO
Cathepsin L of cancer cells has been shown to play an important role in degradation of extracellular matrix for metastasis. In order to reduce cell invasion, cathepsin L propeptide-like proteins which are classified as the I29 family in the MEROPS peptidase database were characterized from Calotropis procera R. Br., rich in cysteine protease. Of 19 candidates, the cloned and expressed recombinant SnuCalCp03-propeptide (rSnuCalCp03-propeptide) showed a low nanomolar Ki value of 2.3 ± 0.2 nM against cathepsin L. A significant inhibition of tumor cell invasion was observed with H1975, HT29, MDA-BM-231, PANC1, and PC3 with a 76, 67, 67, 63, and 79% reduction, respectively, in invasion observed in the presence of 400 nM of the rSnuCalCp03-propeptide. In addition, thermal and pH study showed rSnuCalCp03-propeptide consisting of secondary structures was stable at a broad range of temperatures (30-70 °C) and pH (2-10, except for 5 which is close to the isoelectric point of 5.2).
Assuntos
Calotropis/química , Catepsina L/metabolismo , Clonagem Molecular , Precursores Enzimáticos/metabolismo , Catepsina L/química , Catepsina L/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Precursores Enzimáticos/química , Precursores Enzimáticos/genética , Humanos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Relação Estrutura-AtividadeRESUMO
Genetic aberrations responsible for soft-tissue sarcoma formation in adults are largely unknown, with targeted therapies sorely needed for this complex and heterogeneous family of diseases. Here we report that that the Hippo pathway is deregulated in many soft-tissue sarcomas, resulting in elevated expression of the effector molecule Yes-Associated Protein (YAP). Based on data gathered from human sarcoma patients, a novel autochthonous mouse model, and mechanistic analyses, we determined that YAP-dependent expression of the transcription factor forkhead box M1 (FOXM1) is necessary for cell proliferation/tumorigenesis in a subset of soft-tissue sarcomas. Notably, FOXM1 directly interacts with the YAP transcriptional complex via TEAD1, resulting in coregulation of numerous critical pro-proliferation targets that enhance sarcoma progression. Finally, pharmacologic inhibition of FOXM1 decreases tumor size in vivo, making FOXM1 an attractive therapeutic target for the treatment of some sarcoma subtypes.
Assuntos
Carcinogênese , Fatores de Transcrição Forkhead/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Sarcoma/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/fisiologia , Proteína Forkhead Box M1 , Fatores de Transcrição Forkhead/fisiologia , Via de Sinalização Hippo , Humanos , Fosfoproteínas/metabolismo , Sarcoma/patologia , Fatores de Transcrição , Proteínas de Sinalização YAPRESUMO
The formation of vasculature is essential for tissue maintenance and regeneration. During development, the vasculature forms via the dual processes of vasculogenesis and angiogenesis, and is regulated at multiple levels: from transcriptional hierarchies and protein interactions to inputs from the extracellular environment. Understanding how vascular formation is coordinated in vivo can offer valuable insights into engineering approaches for therapeutic vascularization and angiogenesis, whether by creating new vasculature in vitro or by stimulating neovascularization in vivo. In this Review, we will discuss how the process of vascular development can be used to guide approaches to engineering vasculature. Specifically, we will focus on some of the recently reported approaches to stimulate therapeutic angiogenesis by recreating the embryonic vascular microenvironment using biomaterials for vascular engineering and regeneration.
Assuntos
Vasos Sanguíneos/crescimento & desenvolvimento , Vasos Sanguíneos/fisiologia , Neovascularização Fisiológica , Regeneração , Animais , Materiais Biocompatíveis/farmacologia , Vasos Sanguíneos/efeitos dos fármacos , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Neovascularização Fisiológica/efeitos dos fármacos , Regeneração/efeitos dos fármacos , Engenharia TecidualRESUMO
A new soil ciliate, Pseudonotohymena antarctica n. g., n. sp., from King George Island, Antarctica, is described based on live observation, protargol impregnation, and its 18S rRNA gene. The new genus Pseudonotohymena is morphologically similar to the genus Notohymena Blatterer and Foissner in the following characteristics: 18 fronto-ventral-transverse cirri, a flexible body, undulating membranes, dorsomarginal kineties, and the number of cirri in the marginal rows. However, Pseudonotohymena differs from Notohymena particularly in the dorsal ciliature, that is, in possessing a nonfragmented dorsal kinety (vs. fragmented). In addition, the molecular phylogenetic relationship of the new species differs from that of Notohymena species. On the basis of the morphological features, the genetic data, and morphogenesis, we establish P. antarctica n. g., n. sp. In addition, the cyst morphology of this species is described.
Assuntos
Cilióforos/classificação , RNA Ribossômico 18S/genética , Análise de Sequência de DNA/métodos , Solo/parasitologia , Regiões Antárticas , Cilióforos/genética , Cilióforos/ultraestrutura , DNA de Protozoário/genética , DNA Ribossômico/genética , Filogenia , Especificidade da EspécieRESUMO
This study aimed to develop and evaluate the effects of a sexual abuse prevention mobile application, SAP_MobAPP, for primary school children. Forty-five subjects were trained for 40 minutes once a week. The experimental group received education that utilized the SAP_MobAPP. Control group A received Web based sexual abuse prevention education, while control group B received textbook based sexual abuse prevention education. Effectiveness was verified through a survey on child sexual abuse recognition and avoidance skills administered before and after training. The SAP_MobAPP program improved recognition (awareness) and the child's skills to avoid child sexual abuse situations, and the effects were long-lasting. However, differences between groups were not statistically significant. This study developed a sexual abuse prevention application and verified its effectiveness. Awareness and skills to avoid child sexual abuse after app education increased immediately after training and four weeks later. The SAP_MobAPP could be used for sexual abuse prevention education in schools.
Assuntos
Abuso Sexual na Infância/prevenção & controle , Conhecimentos, Atitudes e Prática em Saúde , Aplicativos Móveis , Serviços de Saúde Escolar , Criança , Feminino , Humanos , Masculino , República da Coreia , Instituições AcadêmicasRESUMO
We discovered a new brackish water oxytrichid Pseudocyrtohymena koreana n. g., n. sp. in South Korea and investigated the new species on the basis of morphology, ontogenesis, and 18S rRNA gene sequences. The new genus has the 18 frontal-ventral-transverse cirri of typical oxytrichids with flexible body, cortical granules, Cyrtohymena undulating membranes (UM), and one left and one right marginal cirral row. Ontogenesis of the new species indicated that dorsal kinety anlage 3 stretches within the parental row without any fragmentations (Urosomoida pattern) and exclusively forms all caudal cirri. The new genus is morphologically similar to Cyrtohymena Foissner, 1989, but has the following distinctive features: (i) caudal cirri absent in dorsal kineties 1 and 2 (vs. present in Cyrtohymena); and (ii) dorsal kinety 3 nonfragmented (vs. fragmented in Cyrtohymena). Further, we collected an additional species Neokeronopsis asiatica Foissner et al. 2010, from King George Island, Antarctica, and the species shares the morphology of UM with Cyrtohymena. Herein, we describe the previously unidentified characteristics of N. asiatica (i.e., cortical granules, body flexibility, contractile vacuole, and 18S rRNA gene sequence). In addition, we obtained two 18S rRNA gene sequences from Cyrtohymena muscorum and Parasterkiella thompsoni to expand samples for phylogenetic analysis. Our 18S rRNA gene tree supports the hypothesis that the Cyrtohymena UM pattern might have evolved several times in hypotrichs (e.g., Neokeronopsidae, Oxytrichinae, and Stylonychinae).
Assuntos
Cilióforos/classificação , Cilióforos/citologia , Organelas/ultraestrutura , Filogenia , Cilióforos/genética , Cilióforos/isolamento & purificação , Análise por Conglomerados , DNA de Protozoário/química , DNA de Protozoário/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Coreia (Geográfico) , Microscopia , Dados de Sequência Molecular , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Água/parasitologiaRESUMO
Many women around the world are suffering from urinary incontinence, defined as the unintentional leakage of urine by external abnormal pressure. Although various kinds of materials have been utilized to treat this disease, therapies that are more effective are still needed for the treatment of urinary incontinence. Here, we present a macro/nanogel composed of in situ forming gelatin-based macrogels and self-assembled heparin-based nanogels, which can serve as an injectable and bioactive bulking material for the treatment of urinary incontinence. The hybrid hydrogels were prepared via enzymatic reaction in the presence of horseradish peroxidase and hydrogen peroxide. Incorporating a growth factor (GF)-loaded heparin nanogel into a gelatin gel matrix enabled the hybrid gel matrix to release GF continuously up to 28 days. Moreover, we demonstrated that the hydrogel composites stimulated the regeneration of the urethral muscle tissue surrounding the urethral wall and promoted the recovery of their biological function when injected in vivo. Thus, the macro/nanohydrogels may provide an advanced therapeutic technique for the treatment of urinary incontinence as well as an application for regenerative medicine.
Assuntos
Materiais Biocompatíveis/administração & dosagem , Polietilenoglicóis/administração & dosagem , Polietilenoimina/administração & dosagem , Uretra/efeitos dos fármacos , Incontinência Urinária/tratamento farmacológico , Animais , Materiais Biocompatíveis/química , Géis/administração & dosagem , Géis/química , Injeções , Nanogéis , Técnicas de Cultura de Órgãos , Polietilenoglicóis/química , Polietilenoimina/química , Ratos , Ratos Sprague-Dawley , Suínos , Resultado do Tratamento , Uretra/patologia , Incontinência Urinária/patologiaRESUMO
Hypoxia plays a critical role in the development and wound healing process, as well as a number of pathological conditions. Here, dextran-based hypoxia-inducible (Dex-HI) hydrogels formed with in situ oxygen consumption via a laccase-medicated reaction are reported. Oxygen levels and gradients were accurately predicted by mathematical simulation. It is demonstrated that Dex-HI hydrogels provide prolonged hypoxic conditions up to 12 h. The Dex-HI hydrogel offers an innovative approach to delineate not only the mechanism by which hypoxia regulates cellular responses, but may facilitate the discovery of new pathways involved in the generation of hypoxic and oxygen gradient environments.
Assuntos
Dextranos/metabolismo , Hidrogéis/metabolismo , Hypoxis/metabolismo , Oxigênio/metabolismo , Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Dextranos/química , Hidrogéis/química , Hypoxis/química , Lacase/química , Lacase/metabolismo , Estrutura Molecular , Oxigênio/química , Consumo de OxigênioRESUMO
Cathepsin L (CTSL), a cysteine cathepsin protease of the papain superfamily, plays a crucial role in cancer progression and metastasis. Dysregulation of CTSL is frequently observed in tumor malignancies, leading to the degradation of extracellular matrix and facilitating epithelial-mesenchymal transition (EMT), a key process in malignant cancer metastasis. This review mainly provides a comprehensive information about recent findings on natural inhibitors targeting CTSL and their anticancer effects, which have emerged as potent anticancer therapeutic agents or metastasis-suppressive adjuvants. Specifically, inhibitors are categorized into small-molecule and macromolecule inhibitors, with a particular emphasis on cathepsin propeptide-type macromolecules. Additionally, the article explores the molecular mechanisms of CTSL involvement in cancer metastasis, highlighting its regulation at transcriptional, translational, post-translational, and epigenetic levels. This work underscores the importance of understanding natural CTSL inhibitors and provides researchers with practical insights to advance the relevant fields and discover novel CTSL-targeting inhibitors from natural sources.
RESUMO
Based on the behavioral theory of the firm, we research how performance feedback influences the formation of CEOs' passion. We articulate that previous performance directly increases CEOs' passion. Specifically, positive affective outcomes (performance above aspiration-level) advance CEOs' positive feelings and motivation toward the domains of success (obsessive passion). For instance, when a firm accomplishes performance objectives, such as reaching sales goals, CEOs' positive feelings toward developing current products would be boosted. Moreover, negative affective outcomes (performance below aspiration-level) would also positively impact CEOs' passion since the CEOs would endeavor to reduce the difference between performance and their aspiration-level. As such, performance feedback is a critical antecedent of CEOs' obsessive passion. In addition, we apply human capital as a moderator between performance feedback and CEOs' obsessive passion. Results based on multiphase survey data from 189 CEOs of Korean small- and medium-sized enterprises indicate that both positive and negative performance feedback positively increase CEOs' obsessive passion. Moreover, human capital negatively moderates the relationship between positive performance feedback and CEOs' obsessive passion and positively moderates the relationship between negative performance feedback and CEOs' obsessive passion.
Assuntos
Emoções , Motivação , Humanos , Retroalimentação , Logro , OcupaçõesRESUMO
Although wound healing is a normal physiological process in the human body, it is often impaired by bacterial infections, ischemia, hypoxia, and excess inflammation, which can lead to chronic and non-healing wounds. Recently, injectable hydrogels with controlled nitric oxide (NO) release behaviour have become potential wound healing therapeutic agents due to their excellent biochemical, mechanical, and biological properties. Here, we proposed novel multifunctional NO-releasing hydrogels that could regulate various wound healing processes, including hemostasis, inflammation, cell proliferation and angiogenesis. By incorporating the copper nanoparticles (NPs) in the network of dual enzymatically crosslinked gelatin hydrogels (GH/Cu), NO was in situ produced via the Cu-catalyzed decomposition of endogenous RSNOs available in the blood, thus resolving the intrinsic shortcomings of NO therapies, such as the short storage and release time, as well as the burst and uncontrollable release modes. We demonstrated that the NO-releasing gelatin hydrogels enhanced the proliferation and migration of endothelial cells, while promoting the M2 (anti-inflammatory) polarization of the macrophage. Furthermore, the effects of NO release on angiogenesis were evaluated using an in vitro tube formation assay and in ovo chicken chorioallantoic membrane (CAM) assay, which revealed that GH/Cu hydrogels could significantly facilitate neovascularization, consistent with the in vivo results. Therefore, we suggested that these hydrogel systems would significantly enhance the wound healing process through the synergistic effects of the hydrogels and NO, and hence could be used as advanced wound dressing materials.
Assuntos
Gelatina , Óxido Nítrico , Humanos , Óxido Nítrico/farmacologia , Gelatina/química , Células Endoteliais , Hidrogéis/química , Cobre/farmacologia , Cicatrização , Anti-Inflamatórios/farmacologia , Movimento Celular , InflamaçãoRESUMO
Myocardial infarction (MI) is treated with stem cell transplantation using various biomaterials and methods, such as stem cell/spheroid injections, cell sheets, and cardiac patches. However, current treatment methods have some limitations, including low stem cell engraftment and poor therapeutic effects. Furthermore, these methods cause secondary damage to heart due to injection and suturing to immobilize them in the heart, inducing side effects. In this study, we developed stem cell spheroid-laden 3-dimensional (3D) patches (S_3DP) with biosealant to treat MI. This 3D patch has dual modules, such as open pockets to directly deliver the spheroids with their paracrine effects and closed pockets to improve the engraft rate by protecting the spheroid from harsh microenvironments. The spheroids formed within S_3DP showed increased viability and expression of angiogenic factors compared to 2-dimensional cultured cells. We also fabricated gelatin-based tissue adhesive biosealants via a thiol-ene reaction and disulfide bond formation. This biosealant showed stronger tissue adhesiveness than commercial fibrin glue. Furthermore, we successfully applied S_3DP using a biosealant in a rat MI model without suturing in vivo, thereby improving cardiac function and reducing heart fibrosis. In summary, S_3DP and biosealant have excellent potential as advanced stem cell therapies with a sutureless approach to MI treatment.
RESUMO
A combination of hydrogels and stem cell spheroids has been used to engineer three-dimensional (3D) osteochondral tissue, but precise zonal control directing cell fate within the hydrogel remains a challenge. In this study, we developed a composite spheroid-laden bilayer hydrogel to imitate osteochondral tissue by spatially controlled differentiation of human adipose-derived stem cells. Meticulous optimization of the spheroid-size and mechanical strength of gelatin methacryloyl (GelMA) hydrogel enables the cells to homogeneously sprout within the hydrogel. Moreover, fibers immobilizing transforming growth factor beta-1 (TGF-ß1) or bone morphogenetic protein-2 (BMP-2) were incorporated within the spheroids, which induced chondrogenic or osteogenic differentiation of cells in general media, respectively. The spheroids-filled GelMA solution was crosslinked to create the bilayer hydrogel, which demonstrated a strong interfacial adhesion between the two layers. The cell sprouting enhanced the adhesion of each hydrogel, demonstrated by increase in tensile strength from 4.8 ± 0.4 to 6.9 ± 1.2 MPa after 14 days of culture. Importantly, the spatially confined delivery of BMP-2 within the spheroids increased mineral deposition and more than threefold enhanced osteogenic genes of cells in the bone layer while the cells induced by TGF-ß1 signals were apparently differentiated into chondrocytes within the cartilage layer. The results suggest that our composite spheroid-laden hydrogel could be used for the biofabrication of osteochondral tissue, which can be applied to engineer other complex tissues by delivery of appropriate biomolecules.
Assuntos
Osteogênese , Fator de Crescimento Transformador beta1 , Humanos , Fator de Crescimento Transformador beta1/farmacologia , Hidrogéis/farmacologia , Engenharia Tecidual/métodos , Diferenciação Celular , Alicerces TeciduaisRESUMO
This study explores the potential for optimizing a sustainable manufacturing process that maintains the essential characteristics of conventional liposomes using food-grade solvents and components. The focus was comparing the physicochemical, morphological, and interfacial properties of liposomes produced with these food-grade ingredients to those made by conventional methods. It was found that there was no significant difference in particle size (195.87 ± 1.40 nm) and ζ-potential (-45.13 ± 0.65 mV) between liposomes made from food-grade and conventional materials. The manufacturing process for liposomes, utilizing food-grade solvents and components, was optimized through the application of Plackett-Burman design and response surface methodology. This approach helped identify key parameters (soy lecithin, ß-sitosterol, W/O ratio) and their optimal values (3.17 g, 0.25 g, 1:2.59). These findings suggest that it is possible to enhance the use of liposomes as an effective and safe delivery system in the food industry, adhering to the strict guidelines set by regulatory agencies.
Assuntos
Lecitinas , Lipossomos , Tamanho da Partícula , Lipossomos/química , Lecitinas/química , Sitosteroides/química , Microfluídica/instrumentação , Glycine max/químicaRESUMO
The removal of Gal80 protein by gene disruption turned into efficient GAL promoter-driven heterologous gene expression under anaerobic alcoholic fermentation of Saccharomyces cerevisiae. Using lipase B from Candida antarctica as a reporter, the relative strength of GAL10 promoter (P(GAL10) ) in Δgal80 mutant that does not require galactose as an inducer was compared to those of ADH1, PDC1, and PGK promoters, which have been known to work well anaerobically in actively fermenting yeast cells under high glucose concentration. P(GAL10) in the Δgal80 mutant showed 0.8-fold (ADH1), fourfold (PDC1), and 50-fold (PGK) in promoter strength.
Assuntos
Proteínas Fúngicas/genética , Engenharia Genética/métodos , Lipase/genética , Regiões Promotoras Genéticas/genética , Proteínas Repressoras/genética , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , Transativadores/genética , Anaerobiose , Etanol/metabolismo , Fermentação , Proteínas Fúngicas/metabolismo , Galactose/metabolismo , Deleção de Genes , Regulação Fúngica da Expressão Gênica , Glucose/metabolismo , Lipase/metabolismo , Organismos Geneticamente Modificados , Saccharomyces cerevisiae/metabolismo , TransgenesRESUMO
Two marine urostylid ciliates, Anteholosticha multicirrata n. sp. and Anteholosticha pulchra (Kahl, 1932) Berger, 2003, were collected from South Korea. These species were identified based on morphology, morphogenesis, and SSU rRNA gene sequence comparison. Anteholosticha multicirrata n. sp. is characterized by the following features: body size 90-125 × 30-45 µm in vivo, shape slender to ellipsoidal in outline, with yellow-greenish cortical granules distributed along and between dorsal kineties and cirri; single contractile vacuole positioned on left at mid-body; three frontal, five to seven frontoterminal, one buccal, one to two pretransverse and four to six transverse cirri; three complete dorsal kineties; one left and one right marginal cirral row; about 117 macronuclear nodules; and three to four micronuclei observed during morphogenesis. In addition, based on the observations of morphogenesis, we found that A. pulchra has pretransverse cirri, which were not described in detail in previous studies. Nuclear small subunit ribosomal RNA (SSU rRNA) gene was used to analyse their phylogenetic relationship, and the gene tree supports that the genus Anteholosticha is a highly polyphyletic group.