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1.
Aquaculture ; 581: 740417, 2024 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-39175872

RESUMO

The lumpfish, Cyclopterus lumpus, holds significant promise as a candidate for large-scale aquaculture production, particularly in its role as a cleaner fish used to manage sea lice infestations in Atlantic salmon Salmo salar farming. Melanomacrophage centers (MMCs) represent polymorphic structures present in the hemolymphopoietic organs of various vertebrates, serving as a widely applicable histological indicator of the fish immune and health status. This study aims to investigate the histochemical characteristics of MMCs within lumpfish livers and to compare MMC density between hatchery-produced (farmed) and wild individuals. Liver samples were collected from 34 lumpfish and subjected to a range of staining techniques, including haematoxylin-eosin, Azan-Mallory's trichrome, Masson-Fontana, Perls-Van Geison, Mallory's hemofuscin, immunohistochemical detection of cytochrome P450 monooxygenase 1 A (CYP1A), and the terminal deoxynucleotidyl transferase-mediated d'UTP nick-end labelling (TUNEL) method. Hepatocytes from hatchery-produced males exhibited notably high lipid content. Additionally, cells showing positive staining with Masson-Fontana, likely associated with the monocyte/macrophage lineage, were identified. Furthermore, small MMCs containing melanin, lipofuscin-ceroids, and ferric ions were detected. While the density of single monocytes/macrophages was markedly higher in hatchery-produced males, no significant discrepancies in MMCs density were observed between wild and hatchery-produced fish, or between males and females of the same origin. The study also revealed the presence of necrotic foci, characterized by hypertrophic hepatocytes positive for both TUNEL and CYP1A staining. These hypertrophic hepatocytes displayed large lipid droplets and pycnotic nuclei, with hatchery-produced males showing a higher numerical density of such foci. In contrast to findings in other fish species, the study found that MMCs did not appear to serve as reliable markers of health status in lumpfish. This conclusion was reached as MMCs density did not exhibit a correlation with necrotic foci or hepatocyte lipid content.

2.
J Virol ; 95(3)2021 01 13.
Artigo em Inglês | MEDLINE | ID: mdl-33148797

RESUMO

Given the projected increase in multidrug-resistant HIV-1, there is an urgent need for development of antiretrovirals that act on virus life cycle stages not targeted by drugs currently in use. Host-targeting compounds are of particular interest because they can offer a high barrier to resistance. Here, we report identification of two related small molecules that inhibit HIV-1 late events, a part of the HIV-1 life cycle for which potent and specific inhibitors are lacking. This chemotype was discovered using cell-free protein synthesis and assembly systems that recapitulate intracellular host-catalyzed viral capsid assembly pathways. These compounds inhibit replication of HIV-1 in human T cell lines and peripheral blood mononuclear cells, and are effective against a primary isolate. They reduce virus production, likely by inhibiting a posttranslational step in HIV-1 Gag assembly. Notably, the compound colocalizes with HIV-1 Gag in situ; however, unexpectedly, selection experiments failed to identify compound-specific resistance mutations in gag or pol, even though known resistance mutations developed upon parallel nelfinavir selection. Thus, we hypothesized that instead of binding to Gag directly, these compounds localize to assembly intermediates, the intracellular multiprotein complexes containing Gag and host factors that form during immature HIV-1 capsid assembly. Indeed, imaging of infected cells shows compound colocalized with two host enzymes found in assembly intermediates, ABCE1 and DDX6, but not two host proteins found in other complexes. While the exact target and mechanism of action of this chemotype remain to be determined, our findings suggest that these compounds represent first-in-class, host-targeting inhibitors of intracellular events in HIV-1 assembly.IMPORTANCE The success of antiretroviral treatment for HIV-1 is at risk of being undermined by the growing problem of drug resistance. Thus, there is a need to identify antiretrovirals that act on viral life cycle stages not targeted by drugs in use, such as the events of HIV-1 Gag assembly. To address this gap, we developed a compound screen that recapitulates the intracellular events of HIV-1 assembly, including virus-host interactions that promote assembly. This effort led to the identification of a new chemotype that inhibits HIV-1 replication at nanomolar concentrations, likely by acting on assembly. This compound colocalized with Gag and two host enzymes that facilitate capsid assembly. However, resistance selection did not result in compound-specific mutations in gag, suggesting that the chemotype does not directly target Gag. We hypothesize that this chemotype represents a first-in-class inhibitor of virus production that acts by targeting a virus-host complex important for HIV-1 Gag assembly.


Assuntos
Antirretrovirais/farmacologia , Infecções por HIV/tratamento farmacológico , HIV-1/efeitos dos fármacos , Leucócitos Mononucleares/efeitos dos fármacos , Bibliotecas de Moléculas Pequenas/farmacologia , Montagem de Vírus/efeitos dos fármacos , Transportadores de Cassetes de Ligação de ATP/metabolismo , RNA Helicases DEAD-box/metabolismo , Infecções por HIV/patologia , Infecções por HIV/virologia , Humanos , Leucócitos Mononucleares/virologia , Proteínas Proto-Oncogênicas/metabolismo , Produtos do Gene gag do Vírus da Imunodeficiência Humana/metabolismo
3.
J Assist Reprod Genet ; 38(10): 2663-2670, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34535847

RESUMO

PURPOSE: A deep learning artificial intelligence (AI) algorithm has been demonstrated to outperform embryologists in identifying euploid embryos destined to implant with an accuracy of 75.3% (1). Our aim was to evaluate the performance of highly trained embryologists in selecting top quality day 5 euploid blastocysts with and without the aid of a deep learning algorithm. MATERIALS AND METHODS: A non-overlapping series of 200 sets of day 5 euploid embryo images with known implantation outcomes was distributed to 17 highly trained embryologists. One embryo in each set was known to have implanted and one failed implantation. They were asked to select which embryo to transfer from each set. The same 200 sets of embryos, with indication of which embryo in each set had been identified by the algorithm as more likely to implant was then distributed. Chi-squared, t-test, and receiver operating curves were performed to compare the embryologist performeance with and without AI. RESULTS: Fourteen embryologists completed both assessments. Embryologists provided with AI results selected successfully implanted embryos in 73.6% of cases compared to 65.5% for those selected using visual assessments alone (p < 0.001). All embryologists improved in their ability to select embryos with the aid of the AI algorithm with a mean percent improvement of 11.1% (range 1.4% to 15.5%). There were no differences in degree of improvement by embryologist level of experience (junior, intermediate, senior). CONCLUSIONS: The incorporation of an AI framework for blastocyst selection enhanced the performance of trained embryologists in identifying PGT-A euploid embryos destined to implant.


Assuntos
Algoritmos , Inteligência Artificial , Blastocisto/classificação , Blastocisto/citologia , Técnicas de Apoio para a Decisão , Implantação do Embrião , Fertilização in vitro/métodos , Adulto , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Gravidez , Taxa de Gravidez , Estudos Retrospectivos
4.
Prehosp Emerg Care ; 24(6): 822-830, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31800335

RESUMO

Background: Dialysis patients are frequently transported to the emergency department (ED) by Emergency Medical Services (EMS) due to acute and severe illness. However, little is known about predictors of first and recurrent transport to the ED (EMS-ED), based on characteristics at the time of dialysis initiation.Methods: We analyzed a cohort of adult (≥18 years) patients affiliated with a large quaternary care center who initiated chronic dialysis from 2009 to 2013 (last follow-up: 2015). Data on patient characteristics at the time of dialysis initiation were linked to regional EMS data. Candidate predictors of first and recurrent EMS-ED transport included comorbid conditions, dialysis characteristics and frailty severity (using the first version of the clinical frailty scale score; CFS). Time to first EMS-ED was analyzed using a multivariable sub-hazards regression model accounting for competing events (transplantation or death). Time to recurrent EMS-ED was analyzed using the Anderson-Gill counting approach, accounting for competing risks.Results: A total of 455 patients were included in the study, 243 (53%) had 1+ EMS-ED events, 90 (20%) never required an EMS-ED at last follow-up, and 69 (15%) and 53 (12%) experienced transplant or death as their first event, respectively. The mean age of the cohort was 62 ± 15 years, 89% were Caucasian, and 35% were female sex. Patients were highly comorbid and 97/381 (25.5%) with available data on frailty severity had a CFS score of ≥5, inclusive of CFS scores ranging from mildly to severely frail. After adjustment, a CFS score of ≥5 (relative to 1-2) was associated with a > 2-fold increase in the risk of first EMS-ED (subdistribution relative hazard; SHR 2.28, 95% confidence interval; CI 1.30-3.98). A history of peripheral vascular disease (SHR 1.43, 95% CI 1.00-2.03) and rheumatologic disease (SHR 1.84, 95% CI 1.00-3.38) was also associated with first EMS-ED. Frailty severity was the only factor associated with recurrent EMS-ED.Conclusion: Patients are at a high risk of EMS-ED after dialysis initiation. Frailty severity (at the time of dialysis initiation) is a strong predictor of first and recurrent EMS-ED and this may be important to guide informed decision making and resource planning for dialysis patients who require EMS.


Assuntos
Serviços Médicos de Emergência , Diálise Renal , Adulto , Idoso , Estudos de Coortes , Serviço Hospitalar de Emergência , Feminino , Fragilidade , Humanos , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais
5.
Ergonomics ; 63(10): 1329-1335, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32588761

RESUMO

This study examined movement economy under load with 1000 g minimalist (MIN) vs. 1600 g traditional (TRD) style boots. Fourteen trained, male participants completed a VO2peak test (46.6 ± 7.3 ml/kg/min) while wearing a 16 kg external load. Treadmill speeds for the running economy (RE) trials were determined by the slowest pace in which participants completed a full stage with a running gait pattern during the VO2peak test. Walking economy (WE) pace was 1.6 km/h slower than RE pace. During the second session, participants completed 5-min exercise bouts at WE and RE pace under load wearing MIN and TRD. There were no differences for any measured variables during WE trials. In contrast, RE (MIN = 2.95 ± 0.28 vs. TRD = 3.04 ± 0.30 L/min; p = .003: Cohen's d = 0.32), respiratory exchange ratio (p < .001), and perceptual measures (p < .05) were all improved while wearing MIN. Practitioner summary: In trained men, 1000 g/pair minimalist style boots (MIN) resulted in improvements of approximately 3% and 5% for running economy and respiratory exchange ratio versus 1600 g/pair traditional boots while wearing a 16 kg kit. Perceptual responses, including comfort, also favoured MIN. These effects were not found at walking pace. Abbreviations: MIN: minimalist style boots; TRD: traditional style boots; RE: running economy; WE: walking economy; ES: effect size; RER: respiratory exchange ratio; HR: heart rate.


Assuntos
Metabolismo Energético/fisiologia , Desenho de Equipamento , Consumo de Oxigênio/fisiologia , Corrida/fisiologia , Sapatos , Caminhada/fisiologia , Adulto , Teste de Esforço , Humanos , Masculino , Militares , Adulto Jovem
6.
Reprod Biomed Online ; 36(6): 658-674, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29656830

RESUMO

This proceedings report presents the outcomes from an international Expert Meeting to establish a consensus on the recommended technical and operational requirements for air quality within modern assisted reproduction technology (ART) laboratories. Topics considered included design and construction of the facility, as well as its heating, ventilation and air conditioning system; control of particulates, micro-organisms (bacteria, fungi and viruses) and volatile organic compounds (VOCs) within critical areas; safe cleaning practices; operational practices to optimize air quality while minimizing physicochemical risks to gametes and embryos (temperature control versus air flow); and appropriate infection-control practices that minimize exposure to VOC. More than 50 consensus points were established under the general headings of assessing site suitability, basic design criteria for new construction, and laboratory commissioning and ongoing VOC management. These consensus points should be considered as aspirational benchmarks for existing ART laboratories, and as guidelines for the construction of new ART laboratories.


Assuntos
Poluição do Ar , Laboratórios/normas , Técnicas de Reprodução Assistida/normas , Poluição do Ar em Ambientes Fechados , Consenso , Monitoramento Ambiental , Humanos
7.
Int J Obes (Lond) ; 40(2): 266-74, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26374448

RESUMO

BACKGROUND/OBJECTIVE: Adiponectin exerts beneficial effects by reducing inflammation and improving lipid metabolism and insulin sensitivity. Although the adiponectin level is lower in obese individuals, whether weight gain reduces adiponectin expression in humans is controversial. We sought to investigate the role of weight gain, and consequent changes in leptin, on altering adiponectin expression in humans. METHODS/RESULTS: Forty-four normal-weight healthy subjects were recruited (mean age 29 years; 14 women) and randomized to either gain 5% of body weight by 8 weeks of overfeeding (n=34) or maintain weight (n=10). Modest weight gain of 3.8±1.2 kg resulted in increased adiponectin level (P=0.03), whereas weight maintenance resulted in no changes in adiponectin. Further, changes in adiponectin correlated positively with changes in leptin (P=0.0085). In-vitro experiments using differentiated human white preadipocytes showed that leptin increased adiponectin mRNA and protein expression, whereas a leptin antagonist had opposite effects. To understand the role of leptin in established obesity, we compared adipose tissue samples obtained from normal-weight versus obese subjects. We noted, first, that leptin activated cellular signaling pathways and increased adiponectin mRNA in the adipose tissue from normal-weight participants, but did not do so in the adipose tissue from obese participants. Second, we noted that obese subjects had increased caveolin-1 expression, which attenuates leptin-dependent increases in adiponectin. CONCLUSIONS: Modest weight gain in healthy individuals is associated with increases in adiponectin levels, which correlate positively with changes in leptin. In vitro, leptin induces adiponectin expression, which is attenuated by increased caveolin-1 expression. In addition, the adipose tissue from obese subjects shows increased caveolin-1 expression and impaired leptin signaling. This leptin signal impairment may prevent concordant increases in adiponectin levels in obese subjects despite their high levels of leptin. Therefore, impaired leptin signaling may contribute to low adiponectin expression in obesity and may provide a target for increasing adiponectin expression, hence improving insulin sensitivity and cardio-metabolic profile in obesity.


Assuntos
Adiponectina/metabolismo , Tecido Adiposo/metabolismo , Envelhecimento/metabolismo , Leptina/metabolismo , Obesidade , Aumento de Peso , Índice de Massa Corporal , Caveolina 1 , Feminino , Humanos , Metabolismo dos Lipídeos , Estudos Longitudinais , Masculino , Obesidade/metabolismo , Obesidade/fisiopatologia , Obesidade/prevenção & controle , Prevalência , Transdução de Sinais , Estados Unidos/epidemiologia , Regulação para Cima
8.
Int J Obes (Lond) ; 39(5): 874-6, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25319743

RESUMO

Subcutaneous adipose tissue can be obtained for research during an elective, clinically indicated operation by standard surgical excision approaches and by needle aspiration in pure research settings. Whether measurements of inflammatory markers and cells from tissues collected in these two different ways are comparable is debatable. We sought to determine whether these two techniques yield systematically different results for measurements of inflammation, cellular senescence and adipose tissue composition. Twelve subjects undergoing surgery participated. At the time of surgery abdominal subcutaneous adipose tissue from adjacent sites was removed by excision and needle aspiration. Stromovascular cell composition (flow cytometry), the number of senescent cells (senescence-associated-ß-galactosidase staining) and interleukin (IL)-6, IL-1, TNF-α and MCP1 mRNA (reverse transcription-PCR) were measured in each sample. We found no statistically significant differences between the two sample-collection approaches for any of the parameters measured. We conclude that these two methods of obtaining adipose tissue do not systematically differ in the results of cytokine mRNA content, cellular senescence or stromovascular cell composition.


Assuntos
Tecido Adiposo/química , Tecido Adiposo/cirurgia , Biópsia por Agulha Fina , Mediadores da Inflamação/análise , Inflamação/metabolismo , Tecido Adiposo/patologia , Biomarcadores/metabolismo , Senescência Celular , Quimiocina CCL2/análise , Feminino , Citometria de Fluxo , Regulação da Expressão Gênica , Humanos , Inflamação/patologia , Interleucina-1/análise , Interleucina-6/análise , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/análise , Fator de Necrose Tumoral alfa/análise
9.
J Virol ; 88(10): 5718-41, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24623418

RESUMO

UNLABELLED: During HIV-1 assembly, Gag polypeptides target to the plasma membrane, where they multimerize to form immature capsids that undergo budding and maturation. Previous mutational analyses identified residues within the Gag matrix (MA) and capsid (CA) domains that are required for immature capsid assembly, and structural studies showed that these residues are clustered on four exposed surfaces in Gag. Exactly when and where the three critical surfaces in CA function during assembly are not known. Here, we analyzed how mutations in these four critical surfaces affect the formation and stability of assembly intermediates in cells expressing the HIV-1 provirus. The resulting temporospatial map reveals that critical MA residues act during membrane targeting, residues in the C-terminal CA subdomain (CA-CTD) dimer interface are needed for the stability of the first membrane-bound assembly intermediate, CA-CTD base residues are necessary for progression past the first membrane-bound intermediate, and residues in the N-terminal CA subdomain (CA-NTD) stabilize the last membrane-bound intermediate. Importantly, we found that all four critical surfaces act while Gag is associated with the cellular facilitators of assembly ABCE1 and DDX6. When correlated with existing structural data, our findings suggest the following model: Gag dimerizes via the CA-CTD dimer interface just before or during membrane targeting, individual CA-CTD hexamers form soon after membrane targeting, and the CA-NTD hexameric lattice forms just prior to capsid release. This model adds an important new dimension to current structural models by proposing the potential order in which key contacts within the immature capsid lattice are made during assembly in cells. IMPORTANCE: While much is known about the structure of the completed HIV-1 immature capsid and domains of its component Gag proteins, less is known about the sequence of events leading to formation of the HIV-1 immature capsid. Here we used biochemical and ultrastructural analyses to generate a temporospatial map showing the precise order in which four critical surfaces in Gag act during immature capsid formation in provirus-expressing cells. Because three of these surfaces make important contacts in the hexameric lattices that are found in the completed immature capsid, these data allow us to propose a model for the sequence of events leading to formation of the hexameric lattices. By providing a dynamic view of when and where critical Gag-Gag contacts form during the assembly process and how those contacts function in the nascent capsid, our study provides novel insights into how an immature capsid is built in infected cells.


Assuntos
Antígenos HIV/metabolismo , Proteína do Núcleo p24 do HIV/metabolismo , HIV-1/fisiologia , Montagem de Vírus , Produtos do Gene gag do Vírus da Imunodeficiência Humana/metabolismo , Transportadores de Cassetes de Ligação de ATP/metabolismo , Capsídeo/metabolismo , RNA Helicases DEAD-box/metabolismo , Análise Mutacional de DNA , Antígenos HIV/genética , Proteína do Núcleo p24 do HIV/genética , HIV-1/genética , Modelos Biológicos , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Ligação Proteica , Proteínas Proto-Oncogênicas/metabolismo , Fatores de Tempo , Produtos do Gene gag do Vírus da Imunodeficiência Humana/genética
10.
Eur Biophys J ; 44(3): 165-70, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25737231

RESUMO

Biophoton intensities depend upon the squared modulus of the electric field. Hence, we first make some general estimates about the inherent electric fields within various biosystems. Generally, these intensities do not follow a simple exponential decay law. After a brief discussion on the inapplicability of a linear rate equation that leads to strict exponential decay, we study other, nonlinear rate equations that have been successfully used for biosystems along with their physical origins when available.


Assuntos
Algoritmos , Eletricidade , Medições Luminescentes/métodos , Fótons , Animais , Humanos
11.
J Virol ; 87(10): 5461-7, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23468488

RESUMO

To overcome the problem of HIV-1 variability, candidate vaccine antigens have been designed to be composed of conserved elements of the HIV-1 proteome. Such candidate vaccines could be improved with a better understanding of both HIV-1 evolutionary constraints and the fitness cost of specific mutations. We evaluated the in vitro fitness cost of 23 mutations engineered in the HIV-1 subtype B Gag-p24 Center-of-Tree (COT) protein through fitness competition assays. While some mutations at conserved sites exacted a high fitness cost, as expected under the assumption that the most conserved residue confers the highest fitness, there was no overall strong relationship between sequence conservation and replicative capacity. By comparing sites that have evolved since the beginning of the epidemic to those that have remain unchanged, we found that sites that have evolved over time were more likely to correspond to HLA-associated sites and that their mutation had limited fitness costs. Our data showed no transcendent link between high conservation and high fitness cost, indicating that merely focusing on conserved segments of HIV-1 would not be sufficient for a successful vaccine strategy. Nonetheless, a subset of sites exacted a high fitness cost upon mutation--these sites have been under selective pressure to change since the beginning of the epidemic but have proved virtually nonmutable and could constitute preferred targets for vaccine design.


Assuntos
Vacinas contra a AIDS/genética , Vacinas contra a AIDS/imunologia , Sequência Conservada , HIV-1/genética , HIV-1/fisiologia , Mutação , Replicação Viral , Evolução Molecular , HIV-1/imunologia , Humanos , Evasão da Resposta Imune , Seleção Genética
12.
Infant Ment Health J ; 35(5): 394-408, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25798491

RESUMO

With a secure foundation in basic research across mammalian species in which fathers participate in the raising of young, novel brain-imaging approaches are outlining a set of consistent brain circuits that regulate paternal thoughts and behaviors in humans. The newest experimental paradigms include increasingly realistic baby-stimuli to provoke paternal cognitions and behaviors with coordinated hormone measures to outline brain networks that regulate motivation, reflexive caring, emotion regulation, and social brain networks with differences and similarities to those found in mothers. In this article, on the father brain, we review all brain-imaging studies on PubMed to date on the human father brain and introduce the topic with a selection of theoretical models and foundational neurohormonal research on animal models in support of the human work. We discuss potentially translatable models for the identification and treatment of paternal mood and father-child relational problems, which could improve infant mental health and developmental trajectories with potentially broad public health importance.


Assuntos
Encéfalo/fisiologia , Pai/psicologia , Saúde Mental , Modelos Animais , Animais , Emoções , Relações Pai-Filho , Feminino , Humanos , Masculino , Neuroimagem , Apoio Social
13.
Am J Transplant ; 13(2): 363-8, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23137119

RESUMO

Obesity is increasingly common before and after liver transplantation (LT), yet optimal management remains unclear. Our aim was to analyze the effectiveness of a multidisciplinary protocol for obese patients requiring LT, including a noninvasive pretransplant weight loss program, and a combined LT plus sleeve gastrectomy (SG) for obese patients who failed to lose weight prior to LT. Since 2006, all patients referred LT with a BMI > 35 were enrolled. There were 37 patients who achieved weight loss and underwent LT alone, and 7 who underwent LT combined with SG. In those who received LT alone, weight gain to BMI > 35 was seen in 21/34, post-LT diabetes (DM) in 12/34, steatosis in 7/34, with 3 deaths plus 3 grafts losses. In patients undergoing the combined procedure, there were no deaths or graft losses. One patient developed a leak from the gastric staple line, and one had excess weight loss. No patients developed post-LT DM or steatosis, and all had substantial weight loss (mean BMI = 29). Noninvasive pretransplant weight loss was achieved by a majority, though weight gain post-LT was common. Combined LT plus SG resulted in effective weight loss and was associated with fewer post-LT metabolic complications. Long-term follow-up is needed.


Assuntos
Derivação Gástrica/métodos , Falência Hepática/terapia , Transplante de Fígado/métodos , Obesidade/cirurgia , Adulto , Idoso , Índice de Massa Corporal , Endoscopia/métodos , Feminino , Gastrectomia/métodos , Humanos , Falência Hepática/complicações , Masculino , Pessoa de Meia-Idade , Obesidade/complicações , Fatores de Risco , Resultado do Tratamento , Redução de Peso
14.
Reprod Biomed Online ; 24(2): 142-7, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22197605

RESUMO

Because media osmolality can impact embryo development, the effect of conditions during microdrop preparation on osmolality was examined. Various sizes of microdrops were prepared under different laboratory conditions. Drops were pipetted directly onto a dish and covered by oil (standard method) or pipetted on the dish, overlaid with oil before removing the underlying media and replaced with fresh media (wash-drop method). Drops were made at 23°C or on a heated stage (37°C) and with or without airflow. Osmolality was assessed at 5 min and 24h. The biological impact of osmolality change was demonstrated by culturing 1-cell mouse embryos in media with varying osmolality. Reduced drop volume, increased temperature and standard method were associated with a significant increase in osmolality at both 5 min and 24h (P-values <0.001, <0.0001 and <0.0001, respectively). There was a significant interaction between airflow, decreased volume, increased temperature and standard method that caused a significant increase in osmolality (40mOsm/kg) compared with controls (P<0.04). There was no significant change in osmolality over time. Mouse embryo development was significantly reduced in media with elevated osmolality (>310mOsm/kg; P<0.05). Procedures in the IVF laboratory can alter osmolality and impact embryo development.


Assuntos
Desenvolvimento Embrionário/fisiologia , Animais , Meios de Cultura , Desenvolvimento Embrionário/efeitos dos fármacos , Camundongos , Concentração Osmolar , Temperatura
15.
Gene Ther ; 18(6): 546-52, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21228882

RESUMO

We use a novel technique that allows for closed recirculation of vector genomes in the cardiac circulation using cardiopulmonary bypass, referred to here as molecular cardiac surgery with recirculating delivery (MCARD). We demonstrate that this platform technology is highly efficient in isolating the heart from the systemic circulation in vivo. Using MCARD, we compare the relative efficacy of single-stranded (ss) adeno-associated virus (AAV)6, ssAAV9 and self-complimentary (sc)AAV6-encoding enhanced green fluorescent protein, driven by the constitutive cytomegalovirus promoter to transduce the ovine myocardium in situ. MCARD allows for the unprecedented delivery of up to 48 green fluorescent protein genome copies per cell globally in the sheep left ventricular (LV) myocardium. We demonstrate that scAAV6-mediated MCARD delivery results in global, cardiac-specific LV gene expression in the ovine heart and provides for considerably more robust and cardiac-specific gene delivery than other available delivery techniques such as intramuscular injection or intracoronary injection; thus, representing a potential, clinically translatable platform for heart failure gene therapy.


Assuntos
Procedimentos Cirúrgicos Cardíacos/métodos , Dependovirus/genética , Técnicas de Transferência de Genes , Terapia Genética/métodos , Vetores Genéticos , Miocárdio , Animais , Ponte Cardiopulmonar , Citomegalovirus , Proteínas de Fluorescência Verde/genética , Miocárdio/metabolismo , Ovinos
16.
Obes Surg ; 30(5): 1952-1960, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32133590

RESUMO

BACKGROUND AND AIMS: To assess safety of the Exilis™ gastric electrical stimulation (GES) system and to investigate whether the settings can be adjusted for comfortable chronic use in subjects with morbid obesity. Gastric emptying and motility and meal intake were evaluated. METHOD: In a multicenter, phase 1, open prospective cohort study, 20 morbidly obese subjects (17 female, mean BMI of 40.8 ± 0.7 kg/m2) were implanted with the Exilis™ system. Amplitude of the Exilis™ system was individually set during titration visits. Subjects underwent two blinded baseline test days (GES ON vs. OFF), after which long-term, monthly follow-up continued for up to 52 weeks. RESULTS: The procedure was safe, and electrical stimulation was well tolerated and comfortable in all subjects. No significant differences in gastric emptying halftime (203 ± 16 vs. 212 ± 14 min, p > 0.05), food intake (713 ± 68 vs. 799 ± 69 kcal, p > 0.05), insulin AUC (2448 ± 347 vs. 2186 ± 204, p > 0.05), and glucose AUC (41 ± 2 vs.41 ± 2, p > 0.05) were found between GES ON and OFF. At week 4, 13, and 26, a significant (p < 0.01) reduction in weight loss was observed but not at week 52. At this time point, the mean excess weight loss (EWL) was 14.2 ± 4.5%. CONCLUSION: Gastric electrical stimulation with the Exilis™ system can be considered as safe. No significant effect on food intake, gastric emptying, or gastric motility was observed. The reduction in weight loss with Exilis™ GES was significant but short lasting. Further electrophysiological research is needed to gain more insight in optimal stimulation parameters and lead localization.


Assuntos
Terapia por Estimulação Elétrica , Obesidade Mórbida , Estimulação Elétrica , Eletrodos Implantados , Feminino , Esvaziamento Gástrico , Humanos , Masculino , Obesidade Mórbida/cirurgia , Estudos Prospectivos
17.
Science ; 237(4812): 280-2, 1987 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-2440105

RESUMO

New blood vessel growth occurs during normal fetal development and in diseases such as cancer and diabetes. The polypeptide angiogenin induces new blood vessel growth in two biological assays and may play a role in the vascular development of the fetus and in the neovascularization that accompanies diseases and wound healing. A complementary DNA probe for human angiogenin was used to examine the tissue distribution of angiogenin messenger RNA (mRNA) in the developing rat and in selected transformed cell lines. Angiogenin mRNA was detected predominantly in adult liver but was also detectable at low levels in other tissues. The expression of the angiogenin gene in rat liver was found to be developmentally regulated; mRNA levels were low in the developing fetus, increased in the neonate, and maximal in the adult. The amount of angiogenin mRNA in human HT-29 colon carcinoma and SK-HEP hepatoma cells was not greater than that in normal rat liver. These results demonstrate that angiogenin is predominantly expressed in adult liver, that the pattern of angiogenin gene expression is not temporally related to vascular development in the rat, and that the transformed cells studied do not contain more angiogenin mRNA than does normal liver. If angiogenin activity is controlled at the transcriptional level, the results of this study suggest that the primary function of angiogenin in vivo may be in processes other than the regulation of vascular growth.


Assuntos
Proteínas de Neoplasias/genética , Ribonuclease Pancreático , Fatores Etários , Animais , Linhagem Celular , Regulação da Expressão Gênica , Humanos , Fígado/fisiologia , Neovascularização Patológica , RNA Mensageiro/genética , Ratos , Distribuição Tecidual
18.
Phys Rev E Stat Nonlin Soft Matter Phys ; 80(1 Pt 2): 016301, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19658800

RESUMO

A water bridge refers to an experimental "flexible cable" made up of pure de-ionized water, which can hang across two supports maintained with a sufficiently large voltage difference. The resulting electric fields within the de-ionized water flexible cable maintain a tension that sustains the water against the downward force of gravity. A detailed calculation of the water bridge tension will be provided in terms of the Maxwell pressure tensor in a dielectric fluid medium. General properties of the dielectric liquid pressure tensor are discussed along with unusual features of dielectric fluid Bernoulli flows in an electric field. The "frictionless" Bernoulli flow is closely analogous to that of a superfluid.

19.
Pharmacol Biochem Behav ; 89(1): 106-15, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18164752

RESUMO

The purpose of the study was to examine the usage of norepinephrine (NE) and dopamine (DA) in the brain when exercising while simultaneously undertaking cognitive tests. Plasma concentrations of the NE metabolite 3-methoxy 4-hydroxyphenylglycol (MHPG) and the DA metabolite homovanillic acid (HVA) showed a linear increase from rest to exercising at 40% and 80% maximum power output (W.max) while simultaneously undertaking cognitive tasks (random number generation (RNG) and response time). Delta plasma concentrations of MHPG and HVA at each exercise intensity while undertaking cognitive tasks and while exercising without cognitive tasks did not differ. Taking blood samples at 0, 1, 3, and 5 min following cessation of exercise did not affect results. Regression correlations showed that delta MHPG and HVA plasma concentrations at the 1 and 3 min sampling times were strong predictors of delta RNG, response time and movement time. Reaction time at 80% W.max significantly increased, while movement time at 80% W.max significantly decreased. It was concluded that these results provide no support for a direct effect of increased catecholamines concentrations on cognitive performance during exercise. The regression data suggest that there is some relationship between exercise, catecholamines concentrations and cognition.


Assuntos
Catecolaminas/fisiologia , Cognição/fisiologia , Exercício Físico/fisiologia , Exercício Físico/psicologia , Adulto , Teste de Esforço , Frequência Cardíaca/fisiologia , Ácido Homovanílico/sangue , Humanos , Masculino , Metoxi-Hidroxifenilglicol/sangue , Desempenho Psicomotor/fisiologia , Tempo de Reação/fisiologia , Análise de Regressão
20.
Indian J Lepr ; 80(2): 175-8, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-19425513

RESUMO

Leprosy is a chronic, infectious disease with varied clinical presentations. The type 2 lepra reaction / erythema nodosum leprosum (ENL) classically involves extremities and face. Besides to the classical lesions of type 2 lepra reaction sometimes rare findings of vesicular lesions with necroticans may be observed. We are reporting a case of lepromatous leprosy with bullous reaction and necroticans involving usual sites along with the unusual involvement of oral cavity, which on first look mimicked necrotizing vasculitis or other bullous disorders. This manifestation of type 2 lepra reaction in oral mucosa should be kept in mind by the physicians while dealing with a case of type 2 lepra reaction.


Assuntos
Eritema Nodoso/patologia , Hanseníase Virchowiana/patologia , Mucosa Bucal/patologia , Adulto , Anti-Inflamatórios/uso terapêutico , Clofazimina/uso terapêutico , Eritema Nodoso/complicações , Feminino , Humanos , Hansenostáticos/uso terapêutico , Hanseníase Virchowiana/complicações , Hanseníase Virchowiana/tratamento farmacológico , Necrose , Prednisolona/uso terapêutico
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