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1.
Chemosphere ; 303(Pt 2): 134993, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35598782

RESUMEN

Aqueous contaminants such as pharmaceuticals, dyes, personal care products, etc., are the common water contaminants that show adverse health effects. Photocatalysis is one of the well-known techniques to treat these water contaminants. Currently, most inorganic photocatalysts show a poor balance between adsorption and photocatalytic activity. In addition, heavy metal pollution and low biosafety are significant concerns in photocatalysis. Thus, environmentally friendly photocatalysts are required to avoid the secondary pollution caused by some inorganic semiconductor-photocatalysts. Organic polymer-based photocatalysts are low-cost, stable, non-toxic, and can utilize visible and NIR light for photocatalysis. In this review, we have discussed polypyrrole as a photocatalyst. Polypyrrole is a conducting organic polymer photocatalyst that is highly stable with high charge mobility and strong binding sites for photocatalytic reactions. Besides these advantages, polypyrrole has limitations, such as high charge recombination due to a small bandgap and poor dispersity. So we have explored the modifications to polypyrrole photocatalysts, such as doping and heterojunctions. Further, we have explained the applications of polypyrrole in photocatalysis as an adsorbent, sensitizer, degradation of pollutants, and energy production. Finally, the future aspects of polypyrrole photocatalysis are also explored to improve the path of future research.


Asunto(s)
Polímeros , Pirroles , Catálisis , Agua
2.
Artículo en Inglés | MEDLINE | ID: mdl-35457636

RESUMEN

Maternity protection is a normative fundamental human right that enables women to combine their productive and reproductive roles, including breastfeeding. The aim of this study is to examine the uptake of Vietnam's maternity protection policy in terms of entitlements and awareness, perceptions, and gaps in implementation through the lens of formally employed women. In this mixed methods study, we interviewed 494 formally employed female workers, among whom 107 were pregnant and 387 were mothers of infants and conducted in-depth interviews with a subset of these women (n = 39). Of the 494 women interviewed, 268 (54.3%) were working in blue-collar jobs and more than 90% were contributing to the public social insurance fund. Among the 387 mothers on paid maternity leave, 51 (13.2%) did not receive cash entitlements during their leave. Among the 182 mothers with infants aged 6-11 months, 30 (16.5%) returned to work before accruing 180 days of maternity leave. Of 121 women who had returned to work, 26 (21.5%) did not receive a one-hour paid break every day to express breastmilk, relax, or breastfeed, and 46 (38.0%) worked the same or more hours per day than before maternity leave. Although most women perceived maternity leave as beneficial for the child's health (92.5%), mother's health (91.5%), family (86.2%), and society (90.7%), fewer women perceived it as beneficial for their income (59.5%), career (46.4%), and employers (30.4%). Not all formally employed women were aware of their maternity protection rights: women were more likely to mention the six-month paid maternity leave (78.7%) and one-hour nursing break (62.3%) than the other nine entitlements (2.0-35.0%). In-depth interviews with pregnant women and mothers of infants supported findings from the quantitative survey. In conclusion, although Vietnam's maternity protection policy helps protect the rights of women and children, our study identified implementation gaps that limit its effectiveness. To ensure that all women and their families can fully benefit from maternity protection, there is a need to increase awareness of the full set of maternity entitlements, strengthen enforcement of existing policies, and expand entitlements to the informal sector.


Asunto(s)
Lactancia Materna , Madres , Niño , Empleo , Femenino , Humanos , Lactante , Sector Informal , Masculino , Embarazo , Vietnam
3.
Chemosphere ; 297: 134074, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35219712

RESUMEN

In the present work, a three-dimensional electrode reactor (3Der) using pallet activated carbon (PAC), as particle electrodes, was investigated to degrade non-biodegradable organic pollutants in pharmaceutical wastewater and steel industry wastewater. The effect of operating parameters, such as pH, electrode distance, O2 flow rate, and current density was investigated. The TOC removal efficiency in 3Der was achieved at the highest mineralization yield of 94.1% after 180 min electrolysis, which was 10-19% higher than the two-dimensional electrode reactor (2Der). The higher performance of the 3Der can be attributed to the indirect and direct oxidation mechanisms. The impact of supporting electrolytes was decreased in order as chloride > nitrate > sulfate. The morphology of sludge and the presence of Fe(OH)3 after Fenton-oxidation were investigated. 3Der system improved biodegradability of pharmaceutical wastewater after electro-Fenton treatment at a PW/SIW ratio of 3:1 (BOD5/COD = 0.6). Hence, the mechanism of 3Der/PAC, as particle electrodes was also proposed. 3Der with PAC particle electrodes using steel industry wastewater as a catalyst is an exciting technique for remediation of organic contaminated pharmaceutical wastewater.


Asunto(s)
Aguas Residuales , Contaminantes Químicos del Agua , Carbón Orgánico , Electrodos , Peróxido de Hidrógeno , Oxidación-Reducción , Preparaciones Farmacéuticas , Acero , Contaminantes Químicos del Agua/análisis
4.
Environ Res ; 209: 112814, 2022 06.
Artículo en Inglés | MEDLINE | ID: mdl-35090874

RESUMEN

The prevalence of global health implications from the COVID-19 pandemic necessitates the innovation and large-scale application of disinfection technologies for contaminated surfaces, air, and wastewater as the significant transmission media of disease. To date, primarily recommended disinfection practices are energy exhausting, chemical driven, and cause severe impact on the environment. The research on advanced oxidation processes has been recognized as promising strategies for disinfection purposes. In particular, semiconductor-based photocatalysis is an effective renewable solar-driven technology that relies on the reactive oxidative species, mainly hydroxyl (•OH) and superoxide (•O2-) radicals, for rupturing the capsid shell of the virus and loss of pathogenicity. However, the limited understanding of critical aspects such as viral photo-inactivation mechanism, rapid virus mutagenicity, and virus viability for a prolonged time restricts the large-scale application of photocatalytic disinfection technology. In this work, fundamentals of photocatalysis disinfection phenomena are addressed with a reviewed remark on the reported literature of semiconductor photocatalysts efficacies against SARS-CoV-2. Furthermore, to validate the photocatalysis process on an industrial scale, we provide updated data on available commercial modalities for an effective virus photo-inactivation process. An elaborative discussion on the long-term challenges and sustainable solutions is suggested to fill in the existing knowledge gaps. We anticipate this review will ignite interest among researchers to pave the way to the photocatalysis process for disinfecting virus-contaminated environments and surfaces for current and future pandemics.


Asunto(s)
COVID-19 , Desinfección , COVID-19/prevención & control , Humanos , Pandemias/prevención & control , SARS-CoV-2 , Aguas Residuales
5.
RSC Adv ; 10(67): 40663-40672, 2020 Nov 09.
Artículo en Inglés | MEDLINE | ID: mdl-35519212

RESUMEN

Understanding the effect of Al doping on CO adsorption at ZnO(101̄0) is crucial for designing a high-performance CO gas sensor. In this work, we investigated the adsorption properties of CO on pristine and Al-doped ZnO(101̄0) by performing DFT+U calculations. It is found that the doping of Al on ZnO(101̄0) induces the semiconductor-to-metal transition and thus enhances the conductance of the substrate. Compared to the pristine ZnO(101̄0), the adsorption energy of CO on the Al-doped surfaces is significantly enhanced since Al doping has the effect of strengthening the adsorption bond. The bonding analysis reveals that CO adsorbs on pristine ZnO(101̄0) via the sole σ-dative donation between the CO HOMO 5σ and the empty states of the Zn cation while π-back donation from filled states of Zn or Al cations to the CO 2π* LUMO is facilitated on the Al-doped surfaces. The π-back donation also results in the red-shift of the CO stretching frequency on the Al-doped surfaces, contrasting to the blue-shift on the pristine surface. The simulated results demonstrate that the doping of Al to a three-fold coordinated site on ZnO(101̄0) is highly beneficial for boosting the performance of the CO gas sensor. Our theoretical investigation provides fundamental insights into the effect of Al doping on the sensing mechanism for CO at the ZnO(101̄0) surface.

6.
Med Image Anal ; 48: 58-74, 2018 08.
Artículo en Inglés | MEDLINE | ID: mdl-29852311

RESUMEN

X-ray image segmentation is an important and crucial step for three-dimensional (3D) bone reconstruction whose final goal remains to increase effectiveness of computer-aided diagnosis, surgery and treatment plannings. However, this segmentation task is rather challenging, particularly when dealing with complicated human structures in the lower limb such as the patella, talus and pelvis. In this work, we present a multi-atlas fusion framework for the automatic segmentation of these complex bone regions from a single X-ray view. The first originality of the proposed approach lies in the use of a (training) dataset of co-registered/pre-segmented X-ray images of these aforementioned bone regions (or multi-atlas) to estimate a collection of superpixels allowing us to take into account all the nonlinear and local variability of bone regions existing in the training dataset and also to simplify the superpixel map pruning process related to our strategy. The second originality is to introduce a novel label propagation step based on the entropy concept for refining the resulting segmentation map into the most likely internal regions to the final consensus segmentation. In this framework, a leave-one-out cross-validation process was performed on 31 manually segmented radiographic image dataset for each bone structure in order to rigorously evaluate the efficiency of the proposed method. The proposed method resulted in more accurate segmentations compared to the probabilistic patch-based label fusion model (PB) and the classical patch-based majority voting fusion scheme (MV) using different registration strategies. Comparison with manual (gold standard) segmentations revealed that the good classification accuracy of our unsupervised segmentation scheme is, respectively, 93.79% for the patella, 88.3% for the talus and 85.02% for the pelvis; a score that falls within the range of accuracy levels of manual segmentations (due to the intra inter/observer variability).


Asunto(s)
Imagenología Tridimensional/métodos , Interpretación de Imagen Radiográfica Asistida por Computador/métodos , Algoritmos , Atlas como Asunto , Entropía , Humanos , Rótula/diagnóstico por imagen , Huesos Pélvicos/diagnóstico por imagen , Astrágalo/diagnóstico por imagen , Rayos X
7.
J Chem Phys ; 141(1): 014708, 2014 Jul 07.
Artículo en Inglés | MEDLINE | ID: mdl-25005304

RESUMEN

The paper presents the results of ab initio study of the opportunities for tuning the band structure, magnetic and transport properties of zigzag graphene nanoribbon (8-ZGNR) on hexagonal boron nitride (h-BN(0001)) semiconductor heterostructure by transverse electric field (E(ext)). This study was performed within the framework of the density functional theory (DFT) using Grimme's (DFT-D2) scheme. We established the critical values of E(ext) for the 8-ZGNR/h-BN(0001) heterostructure, thereby providing for semiconductor-halfmetal transition in one of electron spin configurations. This study also showed that the degeneration in energy of the localized edge states is removed when E(ext) is applied. In ZGNR/h-BN (0001) heterostructure, value of the splitting energy was higher than one in ZGNRs without substrate. We determined the effect of low E(ext) applied to the 8-ZGNR/h-BN (0001) semiconductor heterostructure on the preserved local magnetic moment (LMM) (0.3µ(B)) of edge carbon atoms. The transport properties of the 8-ZGNR/h-BN(0001) semiconductor heterostructure can be controlled using E(ext). In particular, at a critical value of the positive potential, the electron mobility can increase to 7× 10(5) cm(2)/V s or remain at zero in the spin-up and spin-down electron subsystems, respectively. We established that magnetic moments (MMs), band gaps, and carrier mobility can be altered using E(ext). These abilities enable the use of 8-ZGNR/h-BN(0001) semiconductor heterostructure in spintronics.

8.
Nucleic Acids Res ; 29(13): 2789-94, 2001 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-11433024

RESUMEN

The ends of human chromosomes (telomeres) lose up to 200 bp of DNA per cell division. Chromosomal shortening ultimately leads to senescence and death in normal cells. Many human carcinoma lines are immortal in vitro, suggesting that these cells have a mechanism for maintaining the ends of their chromosomes. Telomerase is a ribonucleoprotein complex that synthesizes telomeric DNA onto chromosomes using its RNA component as template. Telomerase activity is found in most tumor cells, but is absent from normal cells. Little is known about how normal human cells repress telomerase (hTERT) gene expression. Mice carrying an E2F-1 null mutation develop a variety of malignant tumors, suggesting that this transcription factor has a tumor suppressor function. To determine mechanisms by which E2F-1 suppresses tumor formation, we examined the role of this transcription factor in regulation of the hTERT promoter in human cells. We identified two putative E2F-1-binding sites proximal to the transcriptional start site of the hTERT promoter. Mutation of these sites produced dramatic increases in promoter activity. Overexpression of E2F-1 but not a mutant E2F-1 repressed hTERT promoter activity in reporter gene assays. This repression was abolished by mutation of the E2F-1-binding sites in the hTERT promoter. Human cancer cell lines stably overexpressing E2F-1 exhibited decreased hTERT mRNA expression and telomerase activity. We conclude that E2F-1 has an atypical function as a transcriptional repressor of the hTERT gene in human cells.


Asunto(s)
Proteínas Portadoras , Proteínas de Ciclo Celular , Proteínas de Unión al ADN/metabolismo , Regulación Enzimológica de la Expresión Génica , Proteínas Represoras/metabolismo , Elementos de Respuesta/genética , Telomerasa/genética , Factores de Transcripción/metabolismo , Secuencia de Bases , Sitios de Unión , Secuencia de Consenso/genética , ADN/genética , ADN/metabolismo , Proteínas de Unión al ADN/genética , Factores de Transcripción E2F , Factor de Transcripción E2F1 , Regulación Neoplásica de la Expresión Génica , Genes Reporteros/genética , Genes Supresores de Tumor/genética , Humanos , Datos de Secuencia Molecular , Mutación/genética , Regiones Promotoras Genéticas/genética , Unión Proteica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Represoras/genética , Proteína 1 de Unión a Retinoblastoma , Telomerasa/metabolismo , Factores de Transcripción/genética , Células Tumorales Cultivadas
9.
Biochim Biophys Acta ; 1518(1-2): 1-6, 2001 Mar 19.
Artículo en Inglés | MEDLINE | ID: mdl-11267653

RESUMEN

The ends of human chromosomes (telomeres) lose up to 200 bp of DNA per cell division. Chromosomal shortening ultimately leads to senescence and death in normal cells. Many human carcinoma lines are immortal in vitro, suggesting that these cells have a mechanism for maintaining the ends of their chromosomes. Telomerase is a ribonucleoprotein complex that synthesizes telomeric DNA onto chromosomes using its RNA component as a template. Recent studies have shown that inactivation of the retinoblastoma gene product pRb and the cyclin dependent kinase inhibitor p16(INK4A) is required for telomerase activity in epithelial cells. We have demonstrated previously that restoration of functional retinoblastoma (Rb) expression is sufficient to downregulate telomerase activity in carcinoma cells. To determine mechanisms by which Rb regulates telomerase expression, we examined the effects of cyclin dependent kinase (cdk) mediated Rb inactivation and the release of E2F-1 on telomerase activity in human carcinoma cells. Overexpression of cdk2 and cdk4 but not a dominant negative cdk2 rescued Rb mediated downregulation of telomerase activity. Overexpression of the cdk regulatory subunit cyclin D1 also rescued telomerase downregulation and p16 expression alone was sufficient to ablate activity. E2F-1 overexpression was sufficient to rescue Rb mediated reduction of telomerase activity, but an E2F-1 mutant defective in DNA and Rb binding activities failed to produce this effect. Tumor tissue from E2F-1 -/- mice was negative for telomerase activity, indicating a key regulatory role for this transcription factor.


Asunto(s)
Quinasas CDC2-CDC28 , Proteínas Portadoras , Proteínas de Ciclo Celular , Proteínas de Unión al ADN , Proteínas Proto-Oncogénicas , Proteína de Retinoblastoma/metabolismo , Telomerasa/metabolismo , Factores de Transcripción/metabolismo , Animales , Ciclina D1/biosíntesis , Ciclina D1/genética , Quinasa 2 Dependiente de la Ciclina , Quinasa 4 Dependiente de la Ciclina , Inhibidor p16 de la Quinasa Dependiente de Ciclina/biosíntesis , Inhibidor p16 de la Quinasa Dependiente de Ciclina/genética , Quinasas Ciclina-Dependientes/antagonistas & inhibidores , Quinasas Ciclina-Dependientes/biosíntesis , Quinasas Ciclina-Dependientes/genética , Factores de Transcripción E2F , Factor de Transcripción E2F1 , Humanos , Ratones , Ratones Noqueados , Proteínas Serina-Treonina Quinasas/biosíntesis , Proteínas Serina-Treonina Quinasas/genética , Proteína de Retinoblastoma/biosíntesis , Proteína de Retinoblastoma/genética , Proteína 1 de Unión a Retinoblastoma , Factor de Transcripción DP1 , Factores de Transcripción/biosíntesis , Factores de Transcripción/genética , Células Tumorales Cultivadas
10.
Br J Oral Maxillofac Surg ; 37(2): 144-6, 1999 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-10371324

RESUMEN

Postanginal septicaemia is a syndrome of anaerobic septicaemia, septic thrombophlebitis of the internal jugular vein, and metastatic infections, that follows a localized infection in the area drained by the large cervical veins. The syndrome was well-known and often fatal in the preantibiotic era. It is now rather rare, presumably as a result of the almost routine use of prophylactic antibiotics. The symptoms are classic, and it should be suspected in any case where septicaemia and metastatic lesions are preceded by a head and neck infection. We report a case that is typical, except that branches of the external jugular vein were thrombosed. To our knowledge this has not been reported previously.


Asunto(s)
Infección Focal Dental , Venas Yugulares , Absceso Periodontal/complicaciones , Infecciones del Sistema Respiratorio/etiología , Sepsis/etiología , Tromboflebitis/etiología , Antibacterianos/uso terapéutico , Dolor en el Pecho/etiología , Infección Focal Dental/microbiología , Humanos , Infecciones por Klebsiella/tratamiento farmacológico , Klebsiella pneumoniae/aislamiento & purificación , Masculino , Persona de Mediana Edad , Cuello/microbiología , Músculos del Cuello/microbiología , Absceso Periodontal/microbiología , Infecciones del Sistema Respiratorio/microbiología , Sepsis/microbiología , Síndrome , Tromboflebitis/microbiología
11.
Biochim Biophys Acta ; 1445(2): 207-15, 1999 May 14.
Artículo en Inglés | MEDLINE | ID: mdl-10320773

RESUMEN

The ends of human chromosomes (telomeres) consist of tandem repeats of the sequence TTAGGG. Telomeres lose up to 200 base pairs of DNA per cell division due to the inability of DNA polymerase to completely replicate the chromosomal ends. Chromosomal shortening ultimately leads to senescence and cell death in normal cells. However, some immortal cells do not lose telomeric sequence during DNA replication. Many human carcinoma lines are immortal in vitro, suggesting that these cells have a mechanism for maintaining the ends of their chromosomes. Telomerase is a ribonucleoprotein complex that synthesizes telomeric DNA onto chromosomes using its RNA component as a template. To elucidate potential mechanisms for telomerase regulation, we tested human squamous cell carcinoma lines (SCCs) for telomerase activity. All SCC lines expressed high levels of telomerase activity. Synchronization in specific cell cycle phases caused marked reduction in telomerase activity in G0 and S, but not in G1 or M. Reduction in telomerase activity correlated with induction of Rb protein in these phases. Overexpression of full length Rb resulted in significant downregulation of telomerase activity. However, expression of an Rb N-terminal oligomerization domain deletion construct, a C-terminal DNA binding domain deletion construct, or a pocket domain mutant failed to downregulate telomerase activity. We concluded that functionally intact Rb was required for cell cycle-dependent downregulation of telomerase activity in SCC lines.


Asunto(s)
Proteína de Retinoblastoma/genética , Telomerasa/genética , Regulación hacia Abajo , Citometría de Flujo , Expresión Génica , Humanos , Mutación , Proteína de Retinoblastoma/biosíntesis , Células Tumorales Cultivadas
12.
J Hum Genet ; 44(1): 43-7, 1999.
Artículo en Inglés | MEDLINE | ID: mdl-9929977

RESUMEN

Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant disorder characterized by tumors of the parathyroid glands, the pancreatic islet cells, and the anterior pituitary. Germline mutations of the MEN1 gene in three independent Japanese cases with MEN1 were analyzed. Case 1 has revealed a 2-bp (TA) insertion at nucleotide position 341 (341insTA) in exon 2, which shifts the reading frame such that the mutant protein has a completely different amino acid sequence from codon 78 to the premature stop codon at 119. In case 2, a nucleotide substitution, i.e., TAG in place of TGG, which encodes tryptophan at codon 198 was identified (nonsense mutation). These mutations were heterozygously present and have not been reported previously. Case 3 showed no mutations in the protein-coding exons and exon-intron junctions of the MEN1 gene by single-strand conformation polymorphism or direct sequencing of the polymerase chain reaction (PCR) fragments. We confirmed the finding that patients with MEN1 carry heterozygous germline mutations in the MEN1 gene, which is compatible with the idea that the MEN1 gene is a tumor suppressor gene. The reason why mutations in the coding region of the MEN1 gene could not be detected by PCR-based analysis in some of the MEN1 patients, e.g. case 3, needs to be clarified further.


Asunto(s)
Mutación de Línea Germinal , Neoplasia Endocrina Múltiple Tipo 1/genética , Adulto , Anciano , Secuencia de Aminoácidos , Secuencia de Bases , Cartilla de ADN/genética , Femenino , Genes Supresores de Tumor , Humanos , Japón , Masculino , Persona de Mediana Edad , Polimorfismo Conformacional Retorcido-Simple
13.
Lasers Surg Med ; 22(1): 14-24, 1998.
Artículo en Inglés | MEDLINE | ID: mdl-9443145

RESUMEN

BACKGROUND AND OBJECTIVE: 5-aminolaevulinic acid (ALA) is a new, promising photosensitizer for PDT of cancer. Subcellular toxicity induced by ALA and light exposure in single cells was studied to elucidate the mechanism of cell damage. STUDY DESIGN/MATERIALS AND METHODS: CPAE, PTK2, and rat neonatal myocardial cells treated with ALA were examined for localization using fluorescence microscopy and for subcellular phototoxicity using 630 nm laser microbeam irradiation of specific subcellular regions. RESULTS: In CPAE and PTK2 cells, a large amount of fluorescence was detected in the peri-nuclear cytoplasm. In rat neonatal myocardial cells, the sensitizer selectively localized in the large mitochondria. In both cell types, there was little phototoxicity when the peripheral cytoplasmic region was exposed, as compared to considerable phototoxicity with exposure of either the perinuclear or nuclear regions. CONCLUSION: Both the CPAE and PTK2 cells demonstrated that the nucleus followed by the perinuclear cytoplasm are the most sensitive cell areas with no sensitivity in the peripheral cytoplasm.


Asunto(s)
Ácido Aminolevulínico/toxicidad , Endotelio Vascular/efectos de los fármacos , Corazón/efectos de los fármacos , Riñón/efectos de los fármacos , Fotoquimioterapia , Fármacos Fotosensibilizantes/toxicidad , Animales , Bovinos , Núcleo Celular/efectos de los fármacos , Células Cultivadas , Citoplasma/efectos de los fármacos , Dipodomys , Endotelio Vascular/citología , Femenino , Riñón/citología , Rayos Láser , Microscopía Fluorescente , Mitocondrias/efectos de los fármacos , Miocardio/citología , Ratas
14.
Lasers Surg Med ; 21(2): 159-65, 1997.
Artículo en Inglés | MEDLINE | ID: mdl-9261793

RESUMEN

BACKGROUND AND OBJECTIVE: Optical trapping is becoming a useful and widespread technique for the micromanipulation of cells and organelles. Giant cell formation following optical trapping was studied to detect the potential adverse effects. STUDY DESIGN/MATERIALS AND METHODS: The nuclei of preselected single CHO cells were exposed to 740 nm and 760 nm laser microbeam generated by a titanium-sapphire tunable laser at 88 and 176 mW and different time exposures. The irradiated single cells were recorded and observed morphologically following exposure. Giant cells were tabulated and photographed. RESULTS: The irradiated cells either failed to divide, or they underwent nuclear proliferation to form giant cells through endoreduplication. CONCLUSION: Giant cells were induced by both 740 nm and 760 nm. The frequency of giant cell formation was higher for the longer time exposures and at the higher power densities. The use of an optical etalon to remove intracavity mode beating and high peak powers of the titanium-sapphire laser caused a significant reduction in the formation of giant cells.


Asunto(s)
Células Gigantes/fisiología , Células Gigantes/efectos de la radiación , Rayos Láser , Animales , Células CHO , División Celular/efectos de la radiación , Núcleo Celular/efectos de la radiación , Células Cultivadas , Cricetinae , Micromanipulación , Dosis de Radiación
15.
J Biol Chem ; 269(20): 14367-70, 1994 May 20.
Artículo en Inglés | MEDLINE | ID: mdl-8182040

RESUMEN

A replication error (RER+) phenotype, characterized by somatic instability in simple repeated sequences, is associated with several types of cancer. To determine if a defect in DNA replication fidelity or repair of replication errors might explain this instability, we compared both processes in cell-free extracts from RER+ endometrial and colorectal cancer cell lines to RER- cell lines. SV40 origin-dependent replication of a microsatellite sequence is highly accurate in cell extracts regardless of their RER phenotype. However, extracts from RER+ cell lines are defective in mismatch repair, while extracts of RER- cell lines are not. Lack of repair was observed when the signal (a nick) for strand-specific repair was either 3' or 5' to the mispair. One colorectal cancer cell line contained deletions in both alleles of the putative mismatch repair gene hMSH2, and one endometrial cancer cell line contained a 4-base pair duplication in one hMSH2 allele. No hMSH2 mutation was detected in the other allele or in the other five RER+ cell lines. Repair was readily detected when each of the defective extracts was mixed with a repair-proficient extract, demonstrating that no trans-acting inhibitor is present. Attempts to complement the repair deficiencies by mixing two different defective extracts identified three combinations that restored repair. The data suggest that: (i) defective repair is associated with colorectal and endometrial cancer and, by extrapolation, with other types of cancer; (ii) mutations in the hMSH2 gene, and possibly other genes, result in defective mismatch repair; (iii) the defect(s) in these lines likely involves pre-incision events or the excision step, but not the incision, polymerization, or ligation steps; and (iv) at least four functional complementation groups for mismatch repair may be involved in human cancer.


Asunto(s)
Neoplasias Colorrectales/genética , Reparación del ADN/genética , Replicación del ADN/genética , ADN Satélite/genética , Neoplasias Endometriales/genética , Línea Celular , Femenino , Prueba de Complementación Genética , Células HeLa , Humanos , Células Tumorales Cultivadas
16.
Biochemistry ; 32(43): 11476-82, 1993 Nov 02.
Artículo en Inglés | MEDLINE | ID: mdl-8218213

RESUMEN

We have previously demonstrated mutagenic bypass of pyrimidine dimers during SV40 origin-dependent replication of UV-irradiated DNA in human cell extracts [Thomas, D. C., & Kunkel, T. A. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 7744-7748]. Here we use two vectors having the origin of replication on opposite sides of a lacZ alpha reporter gene to examine the relative probability of mutagenic translesion synthesis on the leading and lagging strands. Although replication of both vectors is inhibited by UVB irradiation in a dose-dependent manner, the covalently closed DNA products of replication contain T4 endonuclease sensitive sites, indicating that bypass of cyclobutane pyrimidine dimers occurred. At fluences of 70 and 100 J/m2, the mutant frequencies obtained with both vectors are substantially higher than with control DNAs. Sequence analysis of mutants obtained with both vectors reveal three types of mutations at frequencies significantly above those obtained from replication of undamaged DNA. These are C-->T transitions, accounting for about two-thirds of the mutants, a small number of CC-->TT substitutions, and complex mutations. Comparing the distribution of C-->T substitutions in the two spectra permits an estimation of the probability of mutagenic translesion replication of the same sequence when replicated as the leading or lagging strand. The data suggest that the overall average UV-independent C-->T substitution probability per phenotypically detectable dipyrimidine site is the same during leading and lagging strand replication. However, statistically significant differences are observed when the distribution of C-->T substitutions is considered.(ABSTRACT TRUNCATED AT 250 WORDS)


Asunto(s)
Daño del ADN/genética , Replicación del ADN/genética , Mutagénesis , Secuencia de Bases , ADN/efectos de la radiación , Células HeLa , Humanos , Datos de Secuencia Molecular , Probabilidad , Dímeros de Pirimidina , Virus 40 de los Simios , Rayos Ultravioleta
17.
Biochemistry ; 28(12): 5306-10, 1989 Jun 13.
Artículo en Inglés | MEDLINE | ID: mdl-2669973

RESUMEN

The complete amino acid sequence of the subunit of branched-chain amino acid aminotransferase (transaminase B, EC 2.6.1.42) of Salmonella typhimurium was determined. An Escherichia coli recombinant containing the ilvGEDAY gene cluster of Salmonella was used as the source of the hexameric enzyme. The peptide fragments used for sequencing were generated by treatment with trypsin, Staphylococcus aureus V8 protease, endoproteinase Lys-C, and cyanogen bromide. The enzyme subunit contains 308 residues and has a molecular weight of 33,920. To determine the coenzyme-binding site, the pyridoxal 5-phosphate containing enzyme was treated with tritiated sodium borohydride prior to trypsin digestion. Peptide map comparisons with an apoenzyme tryptic digest and monitoring radioactivity incorporation allowed identification of the pyridoxylated peptide, which was then isolated and sequenced. The coenzyme-binding site is the lysyl residue at position 159. The amino acid sequence of Salmonella transaminase B is 97.4% identical with that of Escherichia coli, differing in only eight amino acid positions. Sequence comparisons of transaminase B to other known aminotransferase sequences revealed limited sequence similarity (24-33%) when conserved amino acid substitutions are allowed and alignments were forced to occur on the coenzyme-binding site.


Asunto(s)
Escherichia coli/genética , Vectores Genéticos , Proteínas Recombinantes/biosíntesis , Salmonella typhimurium/enzimología , Transaminasas/biosíntesis , Secuencia de Aminoácidos , Aminoácidos de Cadena Ramificada/análisis , Secuencia de Bases , Sitios de Unión , Escherichia coli/metabolismo , Hidrólisis , Datos de Secuencia Molecular , Fragmentos de Péptidos/análisis , Plásmidos , Proteínas Recombinantes/genética , Salmonella typhimurium/genética , Transaminasas/genética
18.
J Bacteriol ; 171(2): 1196-8, 1989 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-2492508

RESUMEN

Induction of the adaptive response to alkylation damage results in the expression of four genes arranged in three transcriptional units: the ada-alkB operon and the alkA and aidB genes. Adaptive-response induction requires the ada gene product and occurs when cells are treated with methylating agents. In previous studies we noted that aidB, but not alkA or ada-alkB, was induced in the absence of alkylation damage as cells were grown to stationary phase. In this note we present evidence that aidB is induced by anaerobiosis. Thus, aidB is subject to dual regulation by ada-dependent alkylation induction and ada-independent anaerobic induction.


Asunto(s)
Alquilantes/farmacología , Escherichia coli/genética , Genes Bacterianos , Genes , Operón , Transcripción Genética , Alquilación , Anaerobiosis , Inducción Enzimática , Escherichia coli/efectos de los fármacos , Escherichia coli/crecimiento & desarrollo , Genes Reguladores , beta-Galactosidasa/biosíntesis , beta-Galactosidasa/genética
19.
Appl Opt ; 28(17): 3553-5, 1989 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-20555738

RESUMEN

We report an upconversion laser that is pumped by two colors-one red and one near infrared-and emits in the blue-green region.

20.
Anal Chem ; 59(17): 2158-61, 1987 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-19886672

RESUMEN

Single molecules of B-phycoerythrin were detected by laser-induced fluorescence in hydrodynamically focused flows as each molecule transited a focused laser beam. Phycoerythrin is a large protein molecule containing the equivalent of 25 rhodamine-6G chromophores. Single molecule detection is documented by the following: (1 )the number of counts per molecules is in agreement with the expected number, (2) the number of molecules per second is in agreement with the concentration and the flow rate, and (3) the time interval distribution between detected molecules is In agreement with the concentration and the flow rate. The molecular transit time through the 1.1-pL probe volume was 180 micros.

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