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1.
Biomacromolecules ; 24(11): 4718-4730, 2023 11 13.
Artigo em Inglês | MEDLINE | ID: mdl-37651737

RESUMO

High-fidelity preclinical in vitro tissue models can reduce the failure rate of drugs entering clinical trials. Collagen and hyaluronic acid (HA) are major components of the extracellular matrix of many native tissues and affect therapeutic macromolecule diffusion and recovery through tissues. Although collagen and HA are commonly used in tissue engineering, the physical and mechanical properties of these materials are variable and depend highly on processing conditions. In this study, HA was chemically modified and crosslinked via hydrazone bonds to form interpenetrating networks of crosslinked HA (HAX) with collagen (Col). These networks enabled a wide range of mechanical properties, including stiffness and swellability, and microstructures, such as pore morphology and size, that can better recapitulate diverse tissues. We utilized these interpenetrating ColHAX hydrogels as in vitro tissue models to examine macromolecular transport and recovery for early-stage drug screening. Hydrogel formulations with varying collagen and HAX concentrations imparted different gel properties based on the ratio of collagen to HAX. These gels were stable and swelled up to 170% of their original mass, and the storage moduli of the ColHAX gels increased over an order of magnitude by increasing collagen and HA concentration. Interestingly, when HAX concentration was constant and collagen concentration increased, both the pore size and spatial colocalization of collagen and HA increased. HA in the system dominated the ζ-potentials of the gels. The hydrogel and macromolecule properties impacted the mass transport and recovery of lysozyme, ß-lactoglobulin, and bovine serum albumin (BSA) from the ColHAX gels─large molecules were largely impacted by mesh size, whereas small molecules were influenced primarily by electrostatic forces. Overall, the tunable properties demonstrated by the ColHAX hydrogels can be used to mimic different tissues for early-stage assays to understand drug transport and its relationship to matrix properties.


Assuntos
Colágeno , Ácido Hialurônico , Ácido Hialurônico/química , Colágeno/química , Matriz Extracelular/química , Engenharia Tecidual , Hidrogéis/química
2.
Anal Chem ; 94(48): 16538-16542, 2022 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-36413763

RESUMO

The increased use of deep eutectic solvents (DESs) in recent years has been significant and provides new approaches to sample collection and preparation. At the same time, the use of these new solvents to prepare samples can present challenges for subsequent analyses. Common analytical approaches, such as fluorescent labeling, may not be compatible with the solvents. In this work, we explore how effective three traditional fluorescent labels can be at derivatizing amino acids in the most common DESs, formed from choline chloride and ethylene glycol. We demonstrate that the unique solvent characteristics of the DESs still allow for two of the fluorophores, fluorescein isothiocyanate and 5-carboxyfluorescein succinimidyl ester, to effectively label amino acids. Initial optimizations of the reaction conditions demonstrate that we can effectively label both d- and l-amino acids, in solution with concentrations of amino acids down to 4 µM. Capillary electrophoretic separations following this preparation can detect as little as 50 nM. This is possible without removal of any DES from the sample matrix. These results represent the first complete fluorescent labeling reaction in a DES and subsequent capillary electrophoretic separation of the analytes.


Assuntos
Aminoácidos , Solventes Eutéticos Profundos , Solventes/química , Colina , Etilenoglicol/química
3.
Mater Sci Eng R Rep ; 1462021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34483486

RESUMO

Hydrogels have been widely investigated in biomedical fields due to their similar physical and biochemical properties to the extracellular matrix (ECM). Collagen and hyaluronic acid (HA) are the main components of the ECM in many tissues. As a result, hydrogels prepared from collagen and HA hold inherent advantages in mimicking the structure and function of the native ECM. Numerous studies have focused on the development of collagen and HA hydrogels and their biomedical applications. In this extensive review, we provide a summary and analysis of the sources, features, and modifications of collagen and HA. Specifically, we highlight the fabrication, properties, and potential biomedical applications as well as promising commercialization of hydrogels based on these two natural polymers.

5.
Chemistry ; 22(20): 6755-6758, 2016 05 10.
Artigo em Inglês | MEDLINE | ID: mdl-27059428

RESUMO

A series of one-pot, sequential protocols was developed for the synthesis of novel macrocycles bearing α,ß-unsaturated chemotypes. The method highlights a phosphate tether-mediated approach to establish asymmetry, and consecutive one-pot, sequential processes to access the macrocycles with minimal purification procedures. This library amenable strategy provided diverse macrocycles containing α,ß-unsaturated carbon-, sulfur-, or phosphorus-based warheads.


Assuntos
Compostos Macrocíclicos/síntese química , Lactamas Macrocíclicas/síntese química , Naftalenossulfonatos/síntese química , Estereoisomerismo
6.
J Clin Microbiol ; 52(3): 781-9, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24353002

RESUMO

Molecular diagnostic methods based on the detection of mutations conferring drug resistance are promising technologies for rapidly detecting multidrug-/extensively drug-resistant tuberculosis (M/XDR TB), but large studies of mutations as markers of resistance are rare. The Global Consortium for Drug-Resistant TB Diagnostics analyzed 417 Mycobacterium tuberculosis isolates from multinational sites with a high prevalence of drug resistance to determine the sensitivities and specificities of mutations associated with M/XDR TB to inform the development of rapid diagnostic methods. We collected M/XDR TB isolates from regions of high TB burden in India, Moldova, the Philippines, and South Africa. The isolates underwent standardized phenotypic drug susceptibility testing (DST) to isoniazid (INH), rifampin (RIF), moxifloxacin (MOX), ofloxacin (OFX), amikacin (AMK), kanamycin (KAN), and capreomycin (CAP) using MGIT 960 and WHO-recommended critical concentrations. Eight genes (katG, inhA, rpoB, gyrA, gyrB, rrs, eis, and tlyA) were sequenced using Sanger sequencing. Three hundred seventy isolates were INHr, 356 were RIFr, 292 were MOXr/OFXr, 230 were AMKr, 219 were CAPr, and 286 were KANr. Four single nucleotide polymorphisms (SNPs) in katG/inhA had a combined sensitivity of 96% and specificities of 97 to 100% for the detection of INHr. Eleven SNPs in rpoB had a combined sensitivity of 98% for RIFr. Eight SNPs in gyrA codons 88 to 94 had sensitivities of 90% for MOXr/OFXr. The rrs 1401/1484 SNPs had 89 to 90% sensitivity for detecting AMKr/CAPr but 71% sensitivity for KANr. Adding eis promoter SNPs increased the sensitivity to 93% for detecting AMKr and to 91% for detecting KANr. Approximately 30 SNPs in six genes predicted clinically relevant XDR-TB phenotypes with 90 to 98% sensitivity and almost 100% specificity.


Assuntos
Antituberculosos/farmacologia , Farmacorresistência Bacteriana Múltipla , Tuberculose Extensivamente Resistente a Medicamentos/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Mycobacterium tuberculosis/genética , Mutação Puntual , Antituberculosos/uso terapêutico , Proteínas de Bactérias/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Tuberculose Extensivamente Resistente a Medicamentos/microbiologia , Genótipo , Humanos , Índia , Testes de Sensibilidade Microbiana/métodos , Moldávia , Mycobacterium tuberculosis/isolamento & purificação , Fenótipo , Filipinas , Polimorfismo de Nucleotídeo Único , Sensibilidade e Especificidade , Análise de Sequência de DNA , África do Sul
7.
bioRxiv ; 2023 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-37873247

RESUMO

Sea squirts' or tunicates' bodies are composed of cellulose nanofibers and gallol- functionalized proteins. These sea creatures are known to heal their injuries under seawater by forming crosslinks between gallols and functional groups from other proteins in their bodies. Inspired by their wound healing mechanism, herein, we have developed a tissue sealant using zein (a plant-based protein) and tannic acid (gallol-containing polyphenol). Except for fibrin- based sealants, most commercial surgical adhesives, and sealants available today are derived from petroleum products that compromise their biodegradability. They often have complicated and multi-step synthesis processes that ultimately affect their affordability. To overcome this challenge, we ensured that these sea squirt-inspired tissue sealants are bio-based, easily synthesized, and low-cost. The sealants were studied on their own and with a food-grade enzyme transglutaminase. The adhesion performances of the sealants were found to be higher than physiological pressures in seven out of nine different tissue substrates studied here. Their performance was also better than or on par with the FDA-approved fibrin sealant Tisseel. Ex vivo models demonstrate instant sealing of leaking wounds in less than a minute. The sealants were not only cytocompatible but also showed complete wound healing on par with sutures and Tisseel when applied in vivo on skin incisions in rats. Overall, these sea squirt-inspired bio-based sealants show great potential to replace currently available wound closure methods.

8.
J Affect Disord ; 331: 442-451, 2023 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-36963515

RESUMO

BACKGROUND: Caring Contacts can effectively reduce suicide ideation, attempts, and death. In published clinical trials, Caring Contacts were sent by someone who knew the recipient. At scale, Caring Contacts programs rarely introduce the recipient and sender. It is not known whether receiving Caring Contacts from someone unknown is as effective as messages from someone the recipient has met. METHODS: Pragmatic randomized controlled trial comparing Caring Contacts with (CC+) versus without an introductory phone call (CC). Recruitment occurred January-July 2021, with outcomes assessed at 6 months. Participants were primary care patients or healthcare providers/staff reporting adverse mental health outcomes on a qualifying survey. Participants were sent 11 standardized caring text messages over 6 months; when participants replied, they received personalized unscripted responses. CC+ calls were semi-structured. The primary outcome was loneliness (NIH Toolkit). RESULTS: Participants included 331 patients (mean [SD] age: 45.5 [16.4], 78.9 % female) and 335 healthcare providers/staff (mean [SD] age: 40.9 [11.8], 86.6 % female). There were no significant differences in loneliness at 6 months by treatment arm in either stratum. In patients, mean (SD) loneliness was 61.9 (10.7) in CC, and 60.8 (10.3) in CC+, adjusted mean difference of -1.0 (95 % CI: -3.0, 1.0); p-value = 0.31. In providers/staff, mean (SD) loneliness was 61.2 (11) in CC, and 61.3 (11.1) in CC+, adjusted mean difference of 0.2 (95 % CI: -1.8, 2.2); p-value = 0.83. LIMITATIONS: Study population was 93 % white which may limit generalizability. CONCLUSIONS: Including an initial phone call added operational complexity without significantly improving the effectiveness of a Caring Contacts program.


Assuntos
Transtornos Mentais , Envio de Mensagens de Texto , Humanos , Feminino , Pessoa de Meia-Idade , Adulto , Masculino , Solidão , Ideação Suicida , Pessoal de Saúde
9.
ACS Appl Bio Mater ; 5(2): 630-641, 2022 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-35080852

RESUMO

Surgical adhesives can be useful in wound closure because they reduce the risk of infection and pain associated with sutures and staples. However, there are no commercially available surgical adhesives for soft tissue wound closure. To be effective, soft tissue adhesives must be soft and flexible, strongly cohesive and adhesive, biocompatible, and effective in a moist environment. To address these criteria, we draw inspiration from the elasticity and resilience of elastin proteins and the adhesive of marine mussels. We used an elastin-like polypeptide (ELP) for the backbone of our adhesive material due to its elasticity and biocompatibility. A mussel-inspired adhesive molecule, l-3,4-dihydroxyphenylalanine (DOPA), was incorporated into the adhesive to confer wet-setting adhesion. In this study, an ELP named YKV was designed to include tyrosine residues and lysine residues, which contain amine groups. A modified version of YKV, named mYKV, was created through enzymatic conversion of tyrosine residues into DOPA. The ELPs were combined with iron(III) nitrate, sodium periodate, and/or tris(hydroxymethyl)phosphine (THP) cross-linkers to investigate the effect of DOPA- and amine-based cross-linking on adhesion strength and cure time on porcine skin in a warm, humid environment. Incorporation of DOPA into the ELP increased adhesive strength by 2.5 times and reduced failure rates. Iron cross-linkers improved adhesion in the presence of DOPA. THP increased adhesion for all proteins tested even in the absence of DOPA. Using multiple cross-linkers in a single formulation did not significantly improve adhesion. The adhesives with the highest performance (iron nitrate mixed with mYKV and THP mixed with YKV or mYKV) on porcine skin had 10-18 times higher adhesion than a commercial sealant and reached appreciable adhesive strength within 10 min.


Assuntos
Adesivos , Elastina , Adesivos/farmacologia , Animais , Di-Hidroxifenilalanina/química , Compostos Férricos , Ferro/química , Nitratos , Peptídeos/farmacologia , Suínos
10.
J Allied Health ; 51(4): 281-284, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36473219

RESUMO

The skill to answer items successfully in a standardized exam requires a student to master the language in which the exam is written. This implies total mastering of the English language for students with English as a Second Language (ESL). This commentary shares the experience of the Doctor of Physical Therapy (DPT) Program at the Medical Science Campus at the University of Puerto Rico in identifying strategies used by the program to ensure students pass the National Physical Therapy Examination (NPTE). The DPT Program implemented several strategies to improve students' language proficiency to pass the NPTE. Some of the implemented strategies were providing tests in English, developing assignments to encourage students to read text¬books in English, increasing test questions complexity, increasing exposure to computer-based tests, and recommending students to complete NPTE review courses. These strategies seem to improve NPTE passing rates in students with ESL to the level required by the Commission on Accreditation in Physical Therapy Education (CAPTE). Strategies identified in the literature to help ESL students pass national licensure examinations were: increasing mastery of dialect (English proficiency), enhancing vocabulary development, raising comprehension of questions, and supplementing preparatory activities for at-risk students. Therefore, PT programs with non-native English speakers should identify the factors that affect the passing rate at the NPTE and analyze the information to develop specific strategies for these students.


Assuntos
Idioma , Modalidades de Fisioterapia , Humanos
11.
J Am Coll Cardiol ; 80(6): 613-626, 2022 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-35926935

RESUMO

BACKGROUND: Valvular heart disease is an important contributor to cardiovascular morbidity and mortality and remains underdiagnosed. Deep learning analysis of electrocardiography (ECG) may be useful in detecting aortic stenosis (AS), aortic regurgitation (AR), and mitral regurgitation (MR). OBJECTIVES: This study aimed to develop ECG deep learning algorithms to identify moderate or severe AS, AR, and MR alone and in combination. METHODS: A total of 77,163 patients undergoing ECG within 1 year before echocardiography from 2005-2021 were identified and split into train (n = 43,165), validation (n = 12,950), and test sets (n = 21,048; 7.8% with any of AS, AR, or MR). Model performance was assessed using area under the receiver-operating characteristic (AU-ROC) and precision-recall curves. Outside validation was conducted on an independent data set. Test accuracy was modeled using different disease prevalence levels to simulate screening efficacy using the deep learning model. RESULTS: The deep learning algorithm model accuracy was as follows: AS (AU-ROC: 0.88), AR (AU-ROC: 0.77), MR (AU-ROC: 0.83), and any of AS, AR, or MR (AU-ROC: 0.84; sensitivity 78%, specificity 73%) with similar accuracy in external validation. In screening program modeling, test characteristics were dependent on underlying prevalence and selected sensitivity levels. At a prevalence of 7.8%, the positive and negative predictive values were 20% and 97.6%, respectively. CONCLUSIONS: Deep learning analysis of the ECG can accurately detect AS, AR, and MR in this multicenter cohort and may serve as the basis for the development of a valvular heart disease screening program.


Assuntos
Insuficiência da Valva Aórtica , Estenose da Valva Aórtica , Aprendizado Profundo , Doenças das Valvas Cardíacas , Insuficiência da Valva Mitral , Insuficiência da Valva Aórtica/diagnóstico , Estenose da Valva Aórtica/diagnóstico , Eletrocardiografia , Doenças das Valvas Cardíacas/diagnóstico , Doenças das Valvas Cardíacas/epidemiologia , Humanos , Insuficiência da Valva Mitral/diagnóstico , Insuficiência da Valva Mitral/epidemiologia
12.
P R Health Sci J ; 41(2): 74-81, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35704525

RESUMO

OBJECTIVE: To examine the levels of burnout and self-perceived stress in workers at the Puerto Rico Electric Power Authority, who experienced major disasters: Hurricanes Irma and Maria. METHODS: A quantitative descriptive cross-sectional study with a non-probabilistic convenience sample was conducted in workers at the Puerto Rico Electric Power Authority. A structured questionnaire was administered to a sample of 163 eligible participants, aged 21 years and older, who participated voluntarily. Using employees of the electric company, the study examined the relationships between burnout and several characteristics (years of employment, existing health conditions, and coping strategies) both before and after Hurricanes Irma and Maria. Burnout was assessed with Gil-Monte's Spanish Burnout Inventory, and self-perceived stress was assessed with the 14-item Perceived Stress Scale. RESULTS: Before the hurricanes, 16.6% of the workers reported high levels of burnout syndrome, while, after the hurricanes, the proportion increased to one-fifth (20.9%). Prior to the 2 hurricanes, more than one-fourth (23.4%) of the sample reported being extremely stressed; after the hurricanes, that proportion increased to 55%. Factors such as years of employment, counseling, and self-perceived stress showed significant statistical associations (P < .05) with burnout. CONCLUSION: From a public health standpoint, priority should be given to this population, thereby preventing burnout and any other negative effects of the aftermath (i.e., the lengthy response, recovery, and reconstruction) of these kinds of major disaster.


Assuntos
Tempestades Ciclônicas , Desastres , Esgotamento Psicológico , Estudos Transversais , Humanos , Porto Rico/epidemiologia , Estresse Psicológico/epidemiologia
13.
Org Lett ; 18(3): 516-9, 2016 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-26760683

RESUMO

A pot-economical total synthesis of antifungal Sch-725674, 1, is reported. The approach takes advantage of a number of one-pot, sequential transformations, including a phosphate tether-mediated one-pot, sequential RCM/CM/chemoselective hydrogenation protocol, a one-pot tosylation/acrylation sequence, and a one-pot, sequential Finkelstein reaction/Boord olefination/acetonide deprotection procedure to streamline the synthesis route by reducing isolation and purification procedures, thus saving time. Overall, an asymmetric route has been developed that is efficiently accomplished in seven pots from phosphate (S,S)-triene and with minimal purification.


Assuntos
Antifúngicos/síntese química , Macrolídeos/síntese química , Antifúngicos/química , Antifúngicos/farmacologia , Catálise , Hidrogenação , Macrolídeos/química , Macrolídeos/farmacologia , Estrutura Molecular , Organofosfatos , Estereoisomerismo
14.
J Phys Chem B ; 109(43): 20379-86, 2005 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-16853637

RESUMO

The surface reactions of atomic and molecular oxygen with carbon phosphide films have been studied using X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). Carbon phosphide films were produced by ion implantation of trimethylphosphine into polyethylene. Atmospheric oxidation of carbon phosphide films was dominated by phosphorus oxidation and generated a carbon-containing phosphate surface film. This oxidized surface layer acted as an effective diffusion barrier, limiting the depth of phosphorus oxidation within the carbon phosphide film to < 3 nm. The effect of atomic oxygen (AO) exposure on this oxidized carbon phosphide layer was subsequently probed in situ using XPS. Initially AO exposure resulted in a loss of carbon atoms from the surface, but increased the surface concentration of phosphorus atoms as well as the degree of phosphorus oxidation. For more prolonged AO exposures, a highly oxidized phosphate surface layer formed that appeared to be inert toward further AO-mediated erosion. By utilizing phosphorus-containing hydrocarbon thin films, the phosphorus oxides produced during exposure to AO were found to desorb at temperatures >500 K under vacuum conditions. Results from this study suggest that carbon phosphide films can be used as AO-resistant surface coatings on polymers.

15.
J Phys Chem B ; 119(35): 11928-34, 2015 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-26230183

RESUMO

Ultrastable glasses are generated by vapor deposition on substrates heated near the glass transition temperature (Tg), but it is unclear if the remarkable properties of such glasses are present in ultrathin (<100 nm) films. Here, we demonstrate that the moduli of 50 nm thick N,N'-di(1-naphthyl)-N,N'-diphenyl-(1,1'-biphenyl)-4,4'-diamine (NPD) film can be increased from 1.5 to 2.5 GPa by simply increasing the temperature of the substrate during deposition with a maximum in modulus found at T/Tg = 0.94. This maximum in modulus is the same modulus obtained for very thin (<15 nm) NPD films deposited at 295 K (T/Tg = 0.80). However, the modulus of films deposited at this lower temperature abruptly decreases to approximately 1.5 GPa for thicker films; the modulus from deposition at T/Tg = 0.94 is thickness independent. In addition to the thin film modulus, the substrate temperature significantly impacts the water uptake in NPD films. From QCM, the volume fraction of water at equilibrium with nearly saturated water vapor decreases from nearly 4% to less than 1% as the substrate temperature increases from T/Tg = 0.82 to T/Tg = 0.93. The substrate temperature provides a simple route to control mechanical properties and water uptake into vapor-deposited NPD, and these concepts are likely extendable to other organic electronic materials; the increased moduli and decreased water uptake could enable improved performance and lifetime of small molecule glasses for a variety of organic electronic applications.

16.
Emerg Microbes Infect ; 4(7): e42, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26251830

RESUMO

We report the discovery and confirmation of 23 novel mutations with previously undocumented role in isoniazid (INH) drug resistance, in catalase-peroxidase (katG) gene of Mycobacterium tuberculosis (Mtb) isolates. With these mutations, a synonymous mutation in fabG1 (g609a), and two canonical mutations, we were able to explain 98% of the phenotypic resistance observed in 366 clinical Mtb isolates collected from four high tuberculosis (TB)-burden countries: India, Moldova, Philippines, and South Africa. We conducted overlapping targeted and whole-genome sequencing for variant discovery in all clinical isolates with a variety of INH-resistant phenotypes. Our analysis showed that just two canonical mutations (katG 315AGC-ACC and inhA promoter-15C-T) identified 89.5% of resistance phenotypes in our collection. Inclusion of the 23 novel mutations reported here, and the previously documented point mutation in fabG1, increased the sensitivity of these mutations as markers of INH resistance to 98%. Only six (2%) of the 332 resistant isolates in our collection did not harbor one or more of these mutations. The third most prevalent substitution, at inhA promoter position -8, present in 39 resistant isolates, was of no diagnostic significance since it always co-occurred with katG 315. 79% of our isolates harboring novel mutations belong to genetic group 1 indicating a higher tendency for this group to go down an uncommon evolutionary path and evade molecular diagnostics. The results of this study contribute to our understanding of the mechanisms of INH resistance in Mtb isolates that lack the canonical mutations and could improve the sensitivity of next generation molecular diagnostics.


Assuntos
Antituberculosos/farmacologia , Proteínas de Bactérias/genética , Catalase/genética , Farmacorresistência Bacteriana/genética , Isoniazida/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/genética , Humanos , Testes de Sensibilidade Microbiana , Mutação , Mycobacterium tuberculosis/isolamento & purificação , Oxirredutases/genética , Regiões Promotoras Genéticas/genética , Tuberculose/microbiologia
17.
Epigenetics ; 8(1): 79-91, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23235145

RESUMO

Chromatin regulation is a fundamental mechanism underlying stem cell pluripotency, differentiation, and the establishment of cell type-specific gene expression profiles. To examine the role of chromatin regulation in stem cells in vivo, we study regeneration in the freshwater planarian Schmidtea mediterranea. These animals possess a high concentration of pluripotent stem cells, which are capable of restoring any damaged or lost tissues after injury or amputation. Here, we identify the S. mediterranea homologs of the SET1/MLL family of histone methyltransferases and COMPASS and COMPASS-like complex proteins and investigate their role in stem cell function during regeneration. We identified six S. mediterranea homologs of the SET1/MLL family (set1, mll1/2, trr-1, trr-2, mll5-1 and mll5-2), characterized their patterns of expression in the animal, and examined their function by RNAi. All members of this family are expressed in the stem cell population and differentiated tissues. We show that set1, mll1/2, trr-1, and mll5-2 are required for regeneration and that set1, trr-1 and mll5-2 play roles in the regulation of mitosis. Most notably, knockdown of the planarian set1 homolog leads to stem cell depletion. A subset of planarian homologs of COMPASS and COMPASS-like complex proteins are also expressed in stem cells and implicated in regeneration, but the knockdown phenotypes suggest that some complex members also function in other aspects of planarian biology. This work characterizes the function of the SET1/MLL family in the context of planarian regeneration and provides insight into the role of these enzymes in adult stem cell regulation in vivo.


Assuntos
Epigênese Genética , Histona-Lisina N-Metiltransferase/metabolismo , Família Multigênica , Planárias/citologia , Planárias/genética , Células-Tronco/enzimologia , Animais , Diferenciação Celular/genética , Proliferação de Células , Regulação Enzimológica da Expressão Gênica , Técnicas de Silenciamento de Genes , Histona Metiltransferases , Homeostase/genética , Mitose/genética , Fenótipo , Filogenia , Planárias/enzimologia , Interferência de RNA , Regeneração/genética , Células-Tronco/citologia
18.
Rev Iberoam Micol ; 30(4): 267-70, 2013.
Artigo em Espanhol | MEDLINE | ID: mdl-23402832

RESUMO

BACKGROUND: ß-(1,3)(1,6)-D-glucan is fungal cell wall component that has demonstrated immunomodulatory and anti-cancer effects. The (1,3)-ß-glucan synthase is one of the main enzymes involved in its biosynthesis. AIMS: To design primers to partially amplify and characterize the (1,3)-ß-glucan synthase gene and to determine them in Ganoderma lucidum (G. Lucidum) strain CP-132. METHODS: The primers were designed on the basis of homologous genes in other fungi. Then, using the PCR technique, primers were tested using DNA extracted from the G. lucidum strain CP-382. Amplified sequences were compared with those from the GenBank. RESULTS: Three primer pairs were designed; all of them produced amplicons of the expected size. The sequences obtained with primer pairs BGS2113UmF and BGS3097UmR, and BGS547UmF and BGS2113UmR matched with 2 sections of the (1,3)-ß-glucan synthase gene. The deduced amino acid sequences showed high similarity with homologous genes from other fungi, particularly with those of the Agaricomycetes class. CONCLUSIONS: The primer design to partially amplify the (1,3)-ß-glucan synthase gene of G. lucidum using sequences from homologous genes was successful. These primers will allow to characterize this important enzyme in a wide group of fungi.


Assuntos
DNA Fúngico/genética , Proteínas Fúngicas/genética , Glucosiltransferases/genética , Reação em Cadeia da Polimerase/métodos , Reishi/enzimologia , Agaricales/enzimologia , Agaricales/genética , Sequência de Aminoácidos , Sequência de Bases , Primers do DNA , DNA Intergênico/genética , Dados de Sequência Molecular , Peso Molecular , Filogenia , Reishi/genética , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico
19.
NOVA publ. cient ; 15(27): 46-65, ene.-jun. 2017. tab, graf
Artigo em Espanhol | LILACS, COLNAL | ID: biblio-895070

RESUMO

Resumen Objetivo. Se realizó una revisión sobre las características de las fitasas y nitrogenasas de Bacillus spp. y sus opciones de uso como alternativa biofertilizante. El género Bacillus es secretor de proteínas y metabolitos eficientes para el control de plagas y enfermedades, promueve el crecimiento vegetal a través de la solubilización de fósforo y la producción de reguladores de crecimiento como el ácido indol acético; así mismo participa en la fijación de nitrógeno cuando hace parte de consorcios microbianos. Como biofertilizante es una opción amigable para el suelo y el ambiente que da respuesta a la necesidad de implementar la agricultura sostenible.


Abstract Objective. We conducted a review of the characteristics of the phytase and nitrogenase of Bacillus sp. and their potential use as an alternative biological fertilizer. The genus Bacillus is an efficient secretor of proteins and metabolites; to control pests and diseases, promote plant growth through the solubilization of phosphorus and production of growth regulators as acetic indole. Likewise it is involved in nitrogen fixation when it is part of microbial consortia. As biological fertilizer is a friendly option for the soil and the environment that responds to the need to implement sustainable agriculture.


Assuntos
Humanos , Bacillus , 6-Fitase , Pragas da Agricultura , Fertilizantes
20.
J Colloid Interface Sci ; 354(2): 825-31, 2011 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-21145561

RESUMO

Wetting on a corrugated surface that is formed via wrinkling of a hard skin layer formed by UV oxidation (UVO) of a poly(dimethylsiloxane) (PDMS) slab is studied using advancing and receding water contact angle measurements. The amplitude of the wrinkled pattern can be tuned through the pre-strain of the PDMS prior to surface oxidation. These valleys and peaks in the surface topography lead to anisotropic wetting by water droplets. As the droplet advances, the fluid is free to move along the direction parallel to the wrinkles, but the droplet moving orthogonal to the wrinkles encounters energy barriers due to the topography and slip-stick behavior is observed. As the wrinkle amplitude increases, anisotropy in the sessile droplet increases between parallel and perpendicular directions. For the drops receding perpendicular to the wrinkles formed at high strains, the contact angle tends to decrease steadily towards zero as the drop volume decreases, which can result in apparent hysteresis in the contact angle of over 100°. The wrinkled surfaces can exhibit high sessile and advancing contact angles (>115°), but the receding angle in these cases is generally vanishing as the drop is removed. This effect results in micrometer sized drops remaining in the grooves for these highly wrinkled surfaces, while the flat analogous UVO-treated PDMS shows complete removal of all macroscopic water drops under similar conditions. These wetting characteristics should be considered if these wrinkled surfaces are to be utilized in or as microfluidic devices.

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