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1.
Cancer Sci ; 110(2): 629-638, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30457689

RESUMO

Nicotinamide adenine dinucleotide (NAD) exists in an oxidized form (NAD+ ) and a reduced form (NADH). NAD+ plays crucial roles in cancer metabolism, including in cellular signaling, energy production and redox regulation. However, it remains unclear whether NAD(H) pool size (NAD+ and NADH) could be used as biomarker for colon cancer progression. Here, we showed that the NAD(H) pool size and NAD+ /NADH ratio both increased during colorectal cancer (CRC) progression due to activation of the NAD+ salvage pathway mediated by nicotinamide phosphoribosyltransferase (NAMPT). The NAMPT expression was upregulated in adenoma and adenocarcinoma tissues from CRC patients. The NADH fluorescence intensity measured by two-photon excitation fluorescence (TPEF) microscopy was consistently increased in CRC cell lines, azoxymethane/dextran sodium sulfate (AOM/DSS)-induced CRC tissues and tumor tissues from CRC patients. The increases in the NAD(H) pool inhibited the accumulation of excessive reactive oxygen species (ROS) levels and FK866, a specific inhibitor of NAMPT, treatment decreased the CRC nodule size by increasing ROS levels in AOM/DSS mice. Collectively, our results suggest that NAMPT-mediated upregulation of the NAD(H) pool protects cancer cells against detrimental oxidative stress and that detecting NADH fluorescence by TPEF microscopy could be a potential method for monitoring CRC progression.


Assuntos
Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , NAD/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Adenoma/metabolismo , Adenoma/patologia , Animais , Linhagem Celular Tumoral , Colo/metabolismo , Colo/patologia , Progressão da Doença , Células HCT116 , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/fisiologia , Regulação para Cima/fisiologia
2.
J Clin Microbiol ; 57(6)2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30944190

RESUMO

Colonization of the gastrointestinal and genitourinary tracts of pregnant women with group B Streptococcus (GBS) can result in vertical transmission to neonates during labor/delivery. GBS infections in neonates can cause severe complications, such as sepsis, meningitis, and pneumonia. Accurate detection is critical because administration of intrapartum antibiotics can significantly reduce transmission. We compared the clinical sensitivities of three nucleic acid amplification tests (NAATs), the Hologic Panther Fusion GBS, Luminex Aries GBS, and Cepheid Xpert GBS LB assays, to that of the standard of care culture method recommended for GBS screening using 500 vaginal-rectal swab specimens after 18 to 24 h of broth enrichment. We identified 108 positive specimens (21.6%) by culture, while at least 1 of the 3 NAATs was positive for GBS in 155 specimens (31.0%). All 108 specimens positive by culture were also detected by the Panther Fusion assay, while 107/108 (99.1%) were detected by the Cepheid Xpert and Luminex Aries assays. Of the 61 specimens positive by at least 1 NAAT but negative by culture, 24 (39.3%) were positive by all 3 NAATs, suggesting that they represent true positives (TPs). NAATs offer less hands-on time, greater throughput, faster time to result, and potentially greater sensitivity than culture methods, and they should be considered the new gold standard for intrapartum GBS screening.


Assuntos
Técnicas de Tipagem Bacteriana , Técnicas de Amplificação de Ácido Nucleico , Infecções Estreptocócicas/diagnóstico , Infecções Estreptocócicas/microbiologia , Streptococcus agalactiae/classificação , Streptococcus agalactiae/genética , Adulto , Automação Laboratorial , Técnicas de Tipagem Bacteriana/métodos , Técnicas de Tipagem Bacteriana/normas , Feminino , Humanos , Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas de Amplificação de Ácido Nucleico/normas , Gravidez , Diagnóstico Pré-Natal , Reto/microbiologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Vagina/microbiologia , Fluxo de Trabalho
3.
Mayo Clin Proc Innov Qual Outcomes ; 4(1): 21-30, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32055768

RESUMO

OBJECTIVE: To determine whether a single-use stethoscope diaphragm barrier surface remains aseptic when placed on pathogen-contaminated stethoscopes. METHODS: From May 31 to August 5, 2019, we tested 2 separate barriers using 3 different strains of 7 human pathogens, including extended-spectrum ß-lactamase-producing Escherichia coli, methicillin-resistant Staphylococcus aureus, and vancomycin resistant Enterococcus faecium. RESULTS: For all diaphragms with either of the 2 barriers tested, no growth was recorded for any of the pathogens. Stethoscopes with aseptic barriers remained sterile for up to 24 hours. These single-use barriers also provided aseptic surfaces when stethoscope diaphragms were inoculated with human specimens, including saliva, stool, urine, and sputum. CONCLUSION: Disposable aseptic diaphragm barriers may provide robust and efficient solutions to reduce transmission of pathogens via stethoscopes.

4.
Microbiome ; 8(1): 86, 2020 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-32513256

RESUMO

BACKGROUND: Inanimate surfaces within a hospital serve as a reservoir of microbial life that may colonize patients and ultimately result in healthcare associated infections (HAIs). Critically ill patients in intensive care units (ICUs) are particularly vulnerable to HAIs. Little is known about how the microbiome of the ICU is established or what factors influence its evolution over time. A unique opportunity to bridge the knowledge gap into how the ICU microbiome evolves emerged in our health system, where we were able to characterize microbial communities in an established hospital ICU prior to closing for renovations, during renovations, and then after re-opening. RESULTS: We collected swab specimens from ICU bedrails, computer keyboards, and sinks longitudinally at each renovation stage, and analyzed the bacterial compositions on these surfaces by 16S rRNA gene sequencing. Specimens collected before ICU closure had the greatest alpha diversity, while specimens collected after the ICU had been closed for over 300 days had the least. We sampled the ICU during the 45 days after re-opening; however, within that time frame, the alpha diversity never reached pre-closure levels. There were clear and significant differences in microbiota compositions at each renovation stage, which was driven by environmental bacteria after closure and human-associated bacteria after re-opening and before closure. CONCLUSIONS: Overall, we identified significant differences in microbiota diversity and community composition at each renovation stage. These data help to decipher the evolution of the microbiome in the most critical part of the hospital and demonstrate the significant impacts that microbiota from patients and staff have on the evolution of ICU surfaces. Video Abstract.


Assuntos
Biodiversidade , Microbiologia Ambiental , Arquitetura Hospitalar , Unidades de Terapia Intensiva , Microbiota , Bactérias/genética , Arquitetura Hospitalar/estatística & dados numéricos , Microbiota/genética , RNA Ribossômico 16S/genética , Fatores de Tempo
5.
Fertil Steril ; 82(4): 963-5, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15482782

RESUMO

Irradiation of in-vitro-matured bovine oocytes with x-rays of different durations was performed to develop an alternative to conventional mechanical enucleation methods in somatic cell nuclear transfer. No significant difference in embryo development to the blastocyst stage was detected between nonmechanical and mechanical methods, and cytologic analyses of karyotype and microtubule formation showed the potential availability of x-ray irradiation.


Assuntos
Bovinos/embriologia , Fusão Celular/veterinária , Técnicas de Transferência Nuclear , Oócitos/citologia , Animais , Desenvolvimento Embrionário/fisiologia , Feminino , Raios X
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