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1.
Pain Pract ; 2024 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-38624264

RESUMEN

This manuscript is designed to complement the previously published primer on salary structures for new pain physicians. The previous manuscript "Employment Contract Financial Models for the Pain Physician: A Primer" had a goal of increasing understanding of financial models by pain fellows when preparing for contract negotiations. This manuscript illustrates the many equally important considerations of "non-monetary" values that are a significant part of contract negotiation outside of salary. It contributes to the overall education for trainees and pain physicians on benefits and job responsibilities.

2.
Dev Biol ; 471: 76-88, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33326797

RESUMEN

Our lineage tracing studies using multiple Cre mouse lines showed a concurrent labeling of abundant taste bud cells and the underlying connective tissue with a neural crest (NC) origin, warranting a further examination on the issue of whether there is an NC derivation of taste bud cells. In this study, we mapped NC cell lineages in three different models, Sox10-iCreERT2/tdT mouse, GFP+ neural fold transplantation to GFP- chickens, and Sox10-Cre/GFP-RFP zebrafish model. We found that in mice, Sox10-iCreERT2 specifically labels NC cell lineages with a single dose of tamoxifen at E7.5 and that the labeled cells were widely distributed in the connective tissue of the tongue. No labeled cells were found in taste buds or the surrounding epithelium in the postnatal mice. In the GFP+/GFP- chicken chimera model, GFP+ cells migrated extensively to the cranial region of chicken embryos ipsilateral to the surgery side but were absent in taste buds in the base of oral cavity and palate. In zebrafish, Sox10-Cre/GFP-RFP faithfully labeled known NC-derived tissues but did not label taste buds in lower jaw or the barbel. Our data, together with previous findings in axolotl, indicate that taste buds are not derived from NC cells in rodents, birds, amphibians or teleost fish.


Asunto(s)
Linaje de la Célula , Cresta Neural/embriología , Papilas Gustativas/embriología , Animales , Embrión de Pollo , Pollos , Ratones , Ratones Transgénicos , Cresta Neural/citología , Papilas Gustativas/citología , Pez Cebra
3.
Biotechnol Bioeng ; 114(3): 632-644, 2017 03.
Artículo en Inglés | MEDLINE | ID: mdl-27666939

RESUMEN

During antibody dependent cell cytotoxicity (ADCC) the target cells are killed by monocytes and natural killer cells. ADCC is enhanced when the antibody heavy chain's core N-linked glycan lacks the fucose molecule(s). Several strategies have been utilized to generate fully afucosylated antibodies. A commonly used and efficient approach has been knocking out the FUT8 gene of the Chinese hamster ovary (CHO) host cells, which results in expression of antibody molecules with fully afucosylated glycans. However, a major drawback of the FUT8-KO host is the requirement for undertaking two separate cell line development (CLD) efforts in order to obtain both primarily fucosylated and fully afucosylated antibody species for comparative studies in vitro and in vivo. Even more challenging is obtaining primarily fucosylated and FUT8-KO clones with similar enough product quality attributes to ensure that any observed ADCC advantage(s) can be strictly attributed to afucosylation. Here, we report generation and use of a FX knockout (FXKO) CHO host cell line that is capable of expressing antibody molecules with either primarily fucosylated or fully afucosylated glycan profiles with otherwise similar product quality attributes, depending on addition of fucose to the cell culture media. Hence, the FXKO host not only obviates the requirement for undertaking two separate CLD efforts, but it also averts the need for screening many colonies to identify clones with comparable product qualities. Finally, FXKO clones can express antibodies with the desired ratio of primarily fucosylated to afucosylated glycans when fucose is titrated into the production media, to allow achieving intended levels of FcγRIII-binding and ADCC for an antibody. Biotechnol. Bioeng. 2017;114: 632-644. © 2016 Wiley Periodicals, Inc.


Asunto(s)
Anticuerpos/química , Fucosa/metabolismo , Cetona Oxidorreductasas/genética , Ingeniería de Proteínas/métodos , Proteínas Recombinantes/química , Animales , Anticuerpos/genética , Anticuerpos/metabolismo , Células CHO , Sistemas CRISPR-Cas , Cricetinae , Cricetulus , Fucosa/química , Edición Génica , Técnicas de Inactivación de Genes , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
4.
PLoS One ; 19(1): e0296806, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38236914

RESUMEN

Traditional gender and social norms reinforce asymmetrical power relations, increase the risk of experiencing gender-based violence and mediate poor engagement with sexual and reproductive health services. This study explored gender norms and expectations amongst cisgender adolescents in rural KwaZulu-Natal, South Africa. A purposive sample of 29 adolescents aged 16-19 years old were enrolled as part of a longitudinal qualitative study. The current analysis reports on the first round of in-depth interviews, which focused on the role of men and women in their community. A theoretically informed thematic analysis identified three broad themes: 1) Adolescent interpretation and understanding of gender identity, 2) Gendered essentialism and Gender roles (two sub-themes: Young men: Power through providing, and Young women: The domestication process which highlighted that gender roles were defined by being the provider for men, and the successful fulfilment of traditional domestic behaviours amongst women), 3) Gender and fertility highlighted how participants highly valued fertility as affirming of manhood/womanhood. These norms reinforce gender roles that maintain asymmetrical power relations, carrying them over into adulthood. The subtle social pressure to prove fertility could have unintended consequences for driving teenage pregnancy. Structural, gender-based interventions emphasising positive gender-role development in early childhood are needed.


Asunto(s)
Rol de Género , Embarazo en Adolescencia , Preescolar , Embarazo , Humanos , Masculino , Femenino , Adolescente , Adulto Joven , Adulto , Identidad de Género , Sudáfrica , Conducta Sexual
5.
Front Physiol ; 15: 1381548, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38812882

RESUMEN

Background: Coccidiosis outbreaks in susceptible laying hens can significantly decrease egg production and cause substantial economic loss to the egg industry. The supplementation of poultry diets with chemotherapeutic agents is limited due to antimicrobial resistance and residue in poultry meat or processed products. Therefore, alternative strategies to control coccidiosis are needed, and Artemisia annua (AA) might have the potential to be a phytogenic feed additive, an alternative to anticoccidial agents. This study aimed to investigate the effect of the dietary supplementation of powdered AA on the performance and gut health of laying hens infected with coccidiosis by Eimeria spp. Methods: A total of 225 Hy-Line W-36 laying hens at 23 weeks of age were allocated into 5 treatment groups: 1) control (NC), 2) pair-fed (PF) control, 3) challenged control (CC), 4) CC with dietary inclusion of 0.5% AA (0.5AA), and v) CC with dietary inclusion of 1% AA (1AA). The hens in the CC, 0.5AA, and 1AA groups were orally inoculated with sporulated oocysts of Eimeria maxima (12,500), Eimeria tenella (12,500), and Eimeria acervulina (62,500) at week 25. The PF hens received the same amount of feed consumed by the CC hens from 0-14 days post-inoculation (dpi) of Eimeria spp. The performance of the laying hens, including body weight (BW), hen-day egg production (HDEP), feed intake (FI), and feed conversion ratio (FCR), was measured weekly. Additionally, markers of intestinal health, including gut permeability, lesion score, intestinal morphometry, and immune responses, were evaluated at 6, 14, and 21 dpi. Results: At 6 and 14 dpi, laying hens challenged with Eimeria spp. had a lower BW than PF and NC hens (p < 0.0001). Supplementation of 1% AA improved the HDEP by 8.1% compared to CC hens; however, it was still 15.4% lower than that of PF hens (p < 0.0001). The inclusion of 1% AA did not have any beneficial effect on FI; however, the FCR was improved by 0.61 (2.46) than that of CC hens (3.07; p < 0.0001). The inclusion of 1% AA reduced the severity of the intestinal lesions and increased the recovery of intestinal villi (p < 0.05). Additionally, gut permeability was significantly different between the challenged and non-challenged hens; however, among the challenged hens, the inclusion of AA reduced the gut permeability by 29% compared to CC hens (p < 0.0001). Furthermore, the inclusion of 0.5% AA reduced the inflammatory responses in the infected hens. Conclusion: Dietary inclusion of AA partially restored the performance and gut health of the laying hens and modulated their inflammatory immune response following Eimeria infection; however, further studies are needed to better understand the mode of action and effective dosages to improve the gut health without negative impacts on the performance.

6.
Gastroenterology ; 143(2): 469-80, 2012 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-22510200

RESUMEN

BACKGROUND & AIMS: Early embryogenesis involves cell fate decisions that define the body axes and establish pools of progenitor cells. Development does not stop once lineages are specified; cells continue to undergo specific maturation events, and changes in gene expression patterns lead to their unique physiological functions. Secretory pancreatic acinar cells mature postnatally to synthesize large amounts of protein, polarize, and communicate with other cells. The transcription factor MIST1 is expressed by only secretory cells and regulates maturation events. MIST1-deficient acinar cells in mice do not establish apical-basal polarity, properly position zymogen granules, or communicate with adjacent cells, disrupting pancreatic function. We investigated whether MIST1 directly induces and maintains the mature phenotype of acinar cells. METHODS: We analyzed the effects of Cre-mediated expression of Mist1 in adult Mist1-deficient (Mist1(KO)) mice. Pancreatic tissues were collected and analyzed by light and electron microscopy, immunohistochemistry, real-time polymerase chain reaction analysis, and chromatin immunoprecipitation. Primary acini were isolated from mice and analyzed in amylase secretion assays. RESULTS: Induced expression of Mist1 in adult Mist1(KO) mice restored wild-type gene expression patterns in acinar cells. The acinar cells changed phenotypes, establishing apical-basal polarity, increasing the size of zymogen granules, reorganizing the cytoskeletal network, communicating intercellularly (by synthesizing gap junctions), and undergoing exocytosis. CONCLUSIONS: The exocrine pancreas of adult mice can be remodeled by re-expression of the transcription factor MIST1. MIST1 regulates acinar cell maturation and might be used to repair damaged pancreata in patients with pancreatic disorders.


Asunto(s)
Células Acinares/fisiología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Páncreas Exocrino/citología , Células Acinares/metabolismo , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/deficiencia , Biomarcadores/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microscopía Electrónica , Páncreas Exocrino/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Transducción de Señal
7.
J Virol ; 86(7): 3795-808, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22258251

RESUMEN

Although highly active antiretroviral therapy (HAART) has converted HIV into a chronic disease, a reservoir of HIV latently infected resting T cells prevents the eradication of the virus from patients. To achieve eradication, HAART must be combined with drugs that reactivate the dormant viruses. We examined this problem in an established model of HIV postintegration latency by screening a library of small molecules. Initially, we identified eight molecules that reactivated latent HIV. Using them as templates, additional hits were identified by means of similarity-based virtual screening. One of those hits, 8-methoxy-6-methylquinolin-4-ol (MMQO), proved to be useful to reactivate HIV-1 in different cellular models, especially in combination with other known reactivating agents, without causing T-cell activation and with lower toxicity than that of the initial hits. Interestingly, we have established that MMQO produces Jun N-terminal protein kinase (JNK) activation and enhances the T-cell receptor (TCR)/CD3 stimulation of HIV-1 reactivation from latency but inhibits CD3-induced interleukin-2 (IL-2) and tumor necrosis factor alpha (TNF-α) gene transcription. Moreover, MMQO prevents TCR-induced cell cycle progression and proliferation in primary T cells. The present study documents that the combination of biological screening in a cellular model of viral latency with virtual screening is useful for the identification of novel agents able to reactivate HIV-1. Moreover, we set the bases for a hypothetical therapy to reactivate latent HIV by combining MMQO with physiological or pharmacological TCR/CD3 stimulation.


Asunto(s)
Evaluación Preclínica de Medicamentos , Infecciones por VIH/virología , VIH-1/fisiología , Bibliotecas de Moléculas Pequeñas/farmacología , Activación Viral/efectos de los fármacos , Latencia del Virus/efectos de los fármacos , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/virología , Proliferación Celular/efectos de los fármacos , Infecciones por VIH/inmunología , Infecciones por VIH/fisiopatología , VIH-1/efectos de los fármacos , Humanos
8.
Biomolecules ; 13(1)2023 01 12.
Artículo en Inglés | MEDLINE | ID: mdl-36671539

RESUMEN

The current study aimed to monitor the impact of H2O2-induced oxidative stress on avian bone formation during the early stage of embryonic development. Fertilized Cobb broiler eggs were divided into five treatment groups and micro-injected with varying concentrations of H2O2, i.e., control (PBS; 0 nM), 10 nM, 30 nM, 100 nM, and 300 nM, on embryonic day 3, with continued incubation thereafter. The treatment concentrations were selected based on the level of lipid peroxidation and the survival rate of embryo. Embryos were collected at 6 h, 24 h, 48 h, and 72 h post-injection. The mRNA expression levels of apoptotic markers, antioxidant enzymes, and early bone formation gene markers were measured. The results showed that the microinjection of H2O2 altered the expression pattern of antioxidant enzymes' mRNA during early embryogenesis and decreased the expression of COL1A2 and COL2A1 at 6 h and 24 h post-injection. Decreased expression of BMP, BGLAP, and RUNX2 was observed 48 h post-injection. Additionally, a shorter embryo length was observed in the 100 nM and 300 nM H2O2 treatment groups 72 h post-injection. In conclusion, H2O2-induced oxidative stress suppressed the expression of bone formation gene markers, with chronic effects on avian embryonic development.


Asunto(s)
Antioxidantes , Pollos , Animales , Pollos/genética , Antioxidantes/farmacología , Peróxido de Hidrógeno/farmacología , Osteogénesis , Estrés Oxidativo , Desarrollo Embrionario , ARN Mensajero/genética , ARN Mensajero/metabolismo
9.
Poult Sci ; 101(11): 102111, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36081234

RESUMEN

Infection by Salmonella Typhimurium, a food-borne pathogen, can reduce the poultry production efficiency. The objective of this study was to investigate the effects of tannic acid (TA) supplementation on growth performance, Salmonella colonization, gut barrier integrity, serum endotoxin levels, antioxidant capacity, gut health, and immune function in broilers infected with the Salmonella enterica serovar Typhimurium nalidixic acid resistant strain (STNR). A total of 546 one-day-old broilers were arbitrarily allocated into 6 treatments including 1) Sham-challenged control (SCC; birds fed a basal diet and administrated peptone water); 2) Challenged control (CC; birds fed a basal diet and inoculated with 108 STNR); 3) Tannic acid 0.25 (TA0.25; CC + 0.25 g/kg TA); 4) TA0.5 (CC + 0.5 g/kg TA); 5) TA1 (CC + 1 g/kg TA); and 6) TA2 (CC + 2 g/kg TA). On D 7, supplemental TA linearly reduced STNR colonization in the ceca (P < 0.01), and TA1 and TA2 group had significantly lower reduced STNR colonization in the ceca (P < 0.01). On D 7 to 21, average daily gain tended to be linearly increased by supplemental TA (P = 0.097). The serum endotoxin levels were quadratically decreased by supplemental TA on D 21 (P < 0.05). Supplemental TA quadratically increased ileal villus height (VH; P < 0.05), and the TA0.25 group had higher ileal VH compared to the CC group (P < 0.05). Supplemental TA linearly increased percentage of peripheral blood CD8+ T cells on D 18 (P < 0.01). The TA0.5 group had significantly lower lymphocyte numbers compared to the CC groups (P < 0.05). The abundance of monocytes linearly increased with TA supplementation (P < 0.01). Therefore, broilers fed TA had reduced STNR colonization, increased growth performance, decreased serum endotoxin levels, enhanced gut health in the broilers, and stimulated the immune system in broilers infected with STNR. Supplementation of TA (1-2 g/kg) enhanced growth performance and gut health via antimicrobial and immunostimulatory effects in broilers infected with STNR.


Asunto(s)
Enfermedades de las Aves de Corral , Salmonelosis Animal , Animales , Salmonella typhimurium , Pollos , Salmonelosis Animal/tratamiento farmacológico , Salmonelosis Animal/prevención & control , Enfermedades de las Aves de Corral/tratamiento farmacológico , Alimentación Animal/análisis , Taninos/farmacología , Linfocitos T CD8-positivos , Dieta/veterinaria , Antibacterianos/farmacología , Suplementos Dietéticos , Inmunidad , Endotoxinas
10.
Front Physiol ; 12: 637629, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33597896

RESUMEN

A study was conducted to understand the effects of 25-hydroxyvitamin D3 (25OHD) and 1,25-dihydroxyvitamin D3 (1,25OHD) administration on the expression of key genes related to osteogenesis, adipogenesis, myogenesis, and vitamin D3 metabolism in the chicken embryo. A total of 120 fertilized Cobb 500 eggs were used in the current study and were reared under standard incubation conditions. On embryonic day 3 (ED 3), PBS (C), PBS with 40ng 1,25OHD (1,25D-L), 200ng 1,25OHD (1,25D-H), 40ng 25OHD (25D-L), or 200ng 25OHD (25D-H) were injected into the dorsal vein of developing embryos. Whole embryos were harvested at 1, 3, and 6h post-injection for gene expression analyses (n=8). Gene expression for key osteogenesis markers (RUNX2: runt-related transcription factor 2; BMP2: bone morphogenetic protein 2; COL1A2: collagen type I alpha 2 chain; BGLAP: bone gamma-carboxyglutamate protein; SPP1: secreted phosphoprotein 1; and ALP: alkaline phosphatese), adipogenesis markers (PPAR-γ: peroxisome proliferator-activated receptor gamma; FASN: fatty acid synthase; and FABP4: fatty acid binding protein 4), myogenesis markers (MYOG: myogenin; MYOD1: myogenic differentiation 1; and MYF5: myogenic factor 5), and the enzyme responsible for vitamin D3 inactivation (CYP24A1: cytochrome P450 family 24 subfamily A member 1) were measured using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Data were normalized by the ΔΔCT method and analyzed using a one-way ANOVA. Results indicated that at 1h post-injection, no differences were found among treatments. At 3h, the early osteogenesis differentiation marker, ALP, was increased by 1,25D-H and 25D-H, and 25D-H also stimulated the expression of adipogenesis markers (FAPB4 and FASN). In contrast, the expression of myogenesis markers (MYOD1 and MYF5) was suppressed by 25OHD or 1,25OHD treatments, respectively. At 6h, a late osteogenic differentiation marker, SPP1, was increased by 25D-H. MYOD1 and MYF5 were continuously suppressed by 25OHD treatments or 1,25D-H. The evidence of vitamin D3 metabolite retention was assessed by measuring CYP24A1 expression. At 1h, there were no differences in CYP24A1 expression. At 3h, all treatments upregulated CYP24A1 expression relative to control (PBS) embryos. However, at 6h, only the 25D-H group retained higher CYP24A1 expression compared to the other treatments. In conclusion, the results suggested both 1,25OHD and 25OHD induced chicken embryo osteogenesis and adipogenesis, but inhibited myogenesis during early chicken embryo development. The higher dosage of 25OHD showed a possibility of a longer retention time in the embryos.

11.
ACS Pharmacol Transl Sci ; 3(4): 749-758, 2020 Aug 14.
Artículo en Inglés | MEDLINE | ID: mdl-32821883

RESUMEN

As a result of the COVID-19 pandemic, evidence revealed that SARS-CoV-2 infection caused taste loss at a rate higher than that of influenza. ACE2, the entry receptor of SARS-CoV-2, has been identified in the oral epithelium; however, it is unclear at what developmental stage ACE2 expression emerges and whether ACE2 is expressed in taste buds. To identify the specific developmental stage, we analyzed RNA-Seq data from embryonic and newborn mouse oral tissue. We found that robust ACE2 expression was observed in the newborn oral epithelium. In contrast, only extremely low levels, if any, of ACE2 transcripts in the embryonic stage oral tissue were found (E12.5 and E14.5). Analyses of three public scRNA-seq data sets of adult mouse tongue epithelial cells showed that receptors for various viruses were enriched in distinct clusters of tongue epithelial cells. ACE2 was enriched in a subpopulation of epithelial cells in the basal region of nongustatory filiform papillae but not in the taste papillae or taste buds. Expression of ACE2 was detected in a small proportion of type III taste cells. Our results indicate that when applied across species, nongustatory papilla epithelial cells are the prime targets for SARS-CoV-2 infection in the tongue; thus, taste loss in COVID-19 patients is likely not caused by a direct infection of SARS-CoV-2 to taste bud cells. Additionally, fetuses at different stages of development may have distinct susceptibility to SARS-CoV-2 infection.

12.
PLoS Biol ; 3(2): e41, 2005 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-15719057

RESUMEN

The human immunodeficiency virus (HIV) Tat protein is acetylated by the transcriptional coactivator p300, a necessary step in Tat-mediated transactivation. We report here that Tat is deacetylated by human sirtuin 1 (SIRT1), a nicotinamide adenine dinucleotide-dependent class III protein deacetylase in vitro and in vivo. Tat and SIRT1 coimmunoprecipitate and synergistically activate the HIV promoter. Conversely, knockdown of SIRT1 via small interfering RNAs or treatment with a novel small molecule inhibitor of the SIRT1 deacetylase activity inhibit Tat-mediated transactivation of the HIV long terminal repeat. Tat transactivation is defective in SIRT1-null mouse embryonic fibroblasts and can be rescued by expression of SIRT1. These results support a model in which cycles of Tat acetylation and deacetylation regulate HIV transcription. SIRT1 recycles Tat to its unacetylated form and acts as a transcriptional coactivator during Tat transactivation.


Asunto(s)
Productos del Gen tat/metabolismo , VIH/genética , Histona Desacetilasas/genética , Sirtuinas/genética , Transcripción Genética , Acetilación , Secuencia de Bases , Cartilla de ADN , Regulación Viral de la Expresión Génica , Humanos , Datos de Secuencia Molecular , Sirtuina 1 , Productos del Gen tat del Virus de la Inmunodeficiencia Humana
13.
PLoS Genet ; 1(6): e77, 2005 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-16362078

RESUMEN

Sirtuins are a family of phylogenetically conserved nicotinamide adenine dinucleotide-dependent deacetylases that have a firmly established role in aging. Using a simple Saccharomyces cerevisiae yeast heterochromatic derepression assay, we tested a number of environmental chemicals to address the possibility that humans are exposed to sirtuin inhibitors. Here we show that dihydrocoumarin (DHC), a compound found in Melilotus officinalis (sweet clover) that is commonly added to food and cosmetics, disrupted heterochromatic silencing and inhibited yeast Sir2p as well as human SIRT1 deacetylase activity. DHC exposure in the human TK6 lymphoblastoid cell line also caused concentration-dependent increases in p53 acetylation and cytotoxicity. Flow cytometric analysis to detect annexin V binding to phosphatidylserine demonstrated that DHC increased apoptosis more than 3-fold over controls. Thus, DHC inhibits both yeast Sir2p and human SIRT1 deacetylases and increases p53 acetylation and apoptosis, a phenotype associated with senescence and aging. These findings demonstrate that humans are potentially exposed to epigenetic toxicants that inhibit sirtuin deacetylases.


Asunto(s)
Cumarinas/farmacología , Epigénesis Genética , Aromatizantes/farmacología , Silenciador del Gen , Sirtuinas/antagonistas & inhibidores , Envejecimiento , Apoptosis , Línea Celular Tumoral , Senescencia Celular , Proteínas Fúngicas/química , Inhibidores de Histona Desacetilasas , Histona Desacetilasas/genética , Humanos , Fenotipo , Proteínas Reguladoras de Información Silente de Saccharomyces cerevisiae/antagonistas & inhibidores , Proteínas Reguladoras de Información Silente de Saccharomyces cerevisiae/genética , Sirtuina 1 , Sirtuina 2 , Sirtuinas/genética , Proteína p53 Supresora de Tumor/metabolismo
14.
J Nutr Food Sci ; 8(2)2018.
Artículo en Inglés | MEDLINE | ID: mdl-29770259

RESUMEN

Taste is important in guiding nutritive choices and motivating food intake. The sensory organs for taste are the taste buds, that transduce gustatory stimuli into neural signals. It has been reported that chickens have a low taste bud number and thus low taste acuity. However, more recent studies indicate that chickens have a well-developed taste system and the reported number and distribution of taste buds may have been significantly underestimated. Chickens, as a well-established animal model for research, are also the major species of animals in the poultry industry. Thus, a clear understanding of taste organ formation and the effects of taste sensation on nutrition and feeding practices is important for improving livestock production strategies. In this review, we provide an update on recent findings in chicken taste buds and taste sensation indicating that the chicken taste organ is better developed than previously thought and can serve as an ideal system for multidisciplinary studies including organogenesis, regenerative medicine, feeding and nutritional choices.

15.
Sci Rep ; 7(1): 9131, 2017 08 22.
Artículo en Inglés | MEDLINE | ID: mdl-28831098

RESUMEN

RNA-Seq is a powerful tool in transcriptomic profiling of cells and tissues. We recently identified many more taste buds than previously appreciated in chickens using molecular markers to stain oral epithelial sheets of the palate, base of oral cavity, and posterior tongue. In this study, RNA-Seq was performed to understand the transcriptomic architecture of chicken gustatory tissues. Interestingly, taste sensation related genes and many more differentially expressed genes (DEGs) were found between the epithelium and mesenchyme in the base of oral cavity as compared to the palate and posterior tongue. Further RNA-Seq using specifically defined tissues of the base of oral cavity demonstrated that DEGs between gustatory (GE) and non-gustatory epithelium (NGE), and between GE and the underlying mesenchyme (GM) were enriched in multiple GO terms and KEGG pathways, including many biological processes. Well-known genes for taste sensation were highly expressed in the GE. Moreover, genes of signaling components important in organogenesis (Wnt, TGFß/ BMP, FGF, Notch, SHH, Erbb) were differentially expressed between GE and GM. Combined with other features of chicken taste buds, e.g., uniquely patterned array and short turnover cycle, our data suggest that chicken gustatory tissue provides an ideal system for multidisciplinary studies, including organogenesis and regenerative medicine.


Asunto(s)
Pollos/genética , Organogénesis , Análisis de Secuencia de ARN/métodos , Papilas Gustativas/citología , Animales , Embrión de Pollo , Perfilación de la Expresión Génica/métodos , Mesodermo/química , Mesodermo/citología , Especificidad de Órganos , Hueso Paladar/química , Hueso Paladar/citología , Transducción de Señal , Papilas Gustativas/química , Papilas Gustativas/embriología , Lengua/química , Lengua/citología
16.
J Patient Exp ; 4(1): 10-16, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-28725855

RESUMEN

BACKGROUND: Patients who had received surgical services at Bellin Hospital reported anxiety with the surgical flow. This study tested the hypothesis that the introduction of a surgical navigator, someone who guided the patient and their accompanying others throughout the surgical process, would improve patient satisfaction. METHODS: Ambulatory surgical patients were randomized to control and study groups. The study group patients were assigned a surgical navigator. Prior to discharge from the hospital, patients were asked to complete a patient satisfaction survey. RESULTS: The study group had significantly higher mean scores (P value ≤ 0.026), top box scores (P value ≤ 0.021), and positive comments. CONCLUSION: The addition of a surgical navigator to the perioperative process significantly enhanced patient satisfaction in ambulatory surgical patients.

17.
J Med Chem ; 49(25): 7307-16, 2006 Dec 14.
Artículo en Inglés | MEDLINE | ID: mdl-17149860

RESUMEN

NAD+-dependent histone deacetylases, sirtuins, cleave acetyl groups from lysines of histones and other proteins to regulate their activity. Identification of potent selective inhibitors would help to elucidate sirtuin biology and could lead to useful therapeutic agents. NAD+ has an adenosine moiety that is also present in the kinase cofactor ATP. Kinase inhibitors based upon adenosine mimesis may thus also target NAD+-dependent enzymes. We present a systematic approach using adenosine mimics from one cofactor class (kinase inhibitors) as a viable method to generate new lead structures in another cofactor class (sirtuin inhibitors). Our findings have broad implications for medicinal chemistry and specifically for sirtuin inhibitor design. Our results also raise a question as to whether selectivity profiling for kinase inhibitors should be limited to ATP-dependent targets.


Asunto(s)
Adenosina/química , Inhibidores de Histona Desacetilasas , Inhibidores de Proteínas Quinasas/síntesis química , Sirtuinas/antagonistas & inhibidores , Acetilación , Sitios de Unión , Línea Celular Tumoral , Histona Desacetilasas/química , Humanos , Modelos Moleculares , Imitación Molecular , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/farmacología , Sirtuinas/química , Relación Estructura-Actividad , Tubulina (Proteína)/metabolismo
18.
MAbs ; 8(6): 1098-106, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27216702

RESUMEN

For some antibodies intended for use as human therapeutics, reduced effector function is desired to avoid toxicities that might be associated with depletion of target cells. Since effector function(s), including antibody-dependent cell-mediated cytotoxicity (ADCC), require the Fc portion to be glycosylated, reduced ADCC activity antibodies can be obtained through aglycosylation of the human IgG1 isotype. An alternative is to switch to an IgG4 isotype in which the glycosylated antibody is known to have reduced effector function relative to glycosylated IgG1 antibody. ADCC activity of glycosylated IgG1 antibodies is sensitive to the fucosylation status of the Fc glycan, with both in vitro and in vivo ADCC activity increased upon fucose removal ("afucosylation"). The effect of afucosylation on activity of IgG4 antibodies is less well characterized, but it has been shown to increase the in vitro ADCC activity of an anti-CD20 antibody. Here, we show that both in vitro and in vivo activity of anti-CD20 IgG4 isotype antibodies is increased via afucosylation. Using blends of material made in Chinese hamster ovary (CHO) and Fut8KO-CHO cells, we show that ADCC activity of an IgG4 version of an anti-human CD20 antibody is directly proportional to the fucose content. In mice transgenic for human FcγRIIIa, afucosylation of an IgG4 anti-mouse CD20 antibody increases the B cell depletion activity to a level approaching that of the mIgG2a antibody.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Citotoxicidad Celular Dependiente de Anticuerpos , Antígenos CD20/inmunología , Fucosa/química , Inmunoglobulina G/inmunología , Animales , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/genética , Antígenos CD20/genética , Linfocitos B/inmunología , Sangre/inmunología , Células CHO , Cricetinae , Cricetulus , Femenino , Glicosilación , Humanos , Inmunoglobulina G/química , Inmunoglobulina G/genética , Ganglios Linfáticos/inmunología , Depleción Linfocítica , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Bazo/inmunología
19.
J Med Chem ; 47(21): 5235-43, 2004 Oct 07.
Artículo en Inglés | MEDLINE | ID: mdl-15456267

RESUMEN

To probe the steric requirements for deacylation, we synthesized lysine-derived small molecule substrates and examined structure-reactivity relationships with various histone deacetylases. Rat liver, human HeLa, and human recombinant class I and II histone deacetylases (HDACs) as well as human recombinant NAD(+)-dependent SIRT1 (class III enzyme) were used in these studies. A benzyloxycarbonyl substituent on the alpha-amino group yielded the highest conversion rates. Replacing the epsilon-acetyl group with larger lipophilic acyl substituents led to a pronounced decrease in conversion by class I and II enzymes; the class III enzyme displayed a greater tolerance. Incubations with recombinant FLAG-tagged human HDACs 1, 3, and 6 showed a distinct subtype selectivity among small molecule substrates. The subtype selectivity of HDAC inhibitors could be predicted with these substrates and an easily obtainable mixture of HDAC subtypes.


Asunto(s)
Histona Desacetilasas/metabolismo , Lisina/análogos & derivados , Lisina/síntesis química , Animales , Células HeLa , Humanos , Técnicas In Vitro , Hígado/efectos de los fármacos , Hígado/enzimología , Lisina/farmacocinética , Ratas , Proteínas Recombinantes/metabolismo , Sirtuina 1 , Sirtuinas/metabolismo , Estereoisomerismo , Relación Estructura-Actividad , Especificidad por Sustrato
20.
MAbs ; 5(6): 872-81, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23995614

RESUMEN

Knobs-into-holes is a well-validated heterodimerization technology for the third constant domain of an antibody. This technology has been used to produce a monovalent IgG for clinical development (onartuzumab) and multiple bispecific antibodies. The most advanced uses of this approach, however, have been limited to E. coli as an expression host to produce non-glycosylated antibodies. Here, we applied the technology to mammalian host expression systems to produce glycosylated, effector-function competent heterodimeric antibodies. In our mammalian host system, each arm is secreted as a heavy chain-light chain (H-L) fragment with either the knob or hole mutations to allow for preferential heterodimer formation in vitro with low levels of homodimer contaminants. Like full antibodies, the secreted H-L fragments undergo Fc glycosylation in the endoplasmic reticulum. Using a monospecific anti-CD20 antibody, we show that full antibody-dependent cell-mediated cytotoxicity (ADCC) activity can be retained in the context of a knobs-into-holes heterodimer. Because the knobs-into-holes mutations convert the Fc into an asymmetric heterodimer, this technology was further used to systematically explore asymmetric recognition of the Fc. Our results indicate that afucosylation of half the heterodimer is sufficient to produce ADCC-enhancement similar to that observed for a fully afucosylated antibody with wild-type Fc. However, the most dramatic effect on ADCC activity is observed when two carbohydrate chains are present rather than one, regardless of afucosylation state.


Asunto(s)
Formación de Anticuerpos/inmunología , Citotoxicidad Celular Dependiente de Anticuerpos/inmunología , Animales , Células CHO , Línea Celular , Supervivencia Celular/inmunología , Células Cultivadas , Cricetulus , Dimerización , Fucosa/metabolismo , Glicosilación , Humanos , Ingeniería de Proteínas
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