RESUMEN
Aside from abasic sites and ribonucleotides, the DNA adduct N7-methyl deoxyguanosine (N7 -CH3 dG) is one of the most abundant lesions in mammalian DNA. Because N7 -CH3 dG is unstable, leading to deglycosylation and ring-opening, its miscoding potential is not well-understood. Here, we employed a 2'-fluoro isostere approach to synthesize an oligonucleotide containing an analog of this lesion (N7 -CH3 2'-F dG) and examined its miscoding potential with four Y-family translesion synthesis DNA polymerases (pols): human pol (hpol) η, hpol κ, and hpol ι and Dpo4 from the archaeal thermophile Sulfolobus solfataricus We found that hpol η and Dpo4 can bypass the N7 -CH3 2'-F dG adduct, albeit with some stalling, but hpol κ is strongly blocked at this lesion site, whereas hpol ι showed no distinction with the lesion and the control templates. hpol η yielded the highest level of misincorporation opposite the adduct by inserting dATP or dTTP. Moreover, hpol η did not extend well past an N7-CH3 2'-F dG:dT mispair. MS-based sequence analysis confirmed that hpol η catalyzes mainly error-free incorporation of dC, with misincorporation of dA and dG in 5-10% of products. We conclude that N7-CH3 2'-F dG and, by inference, N7-CH3 dG have miscoding and mutagenic potential. The level of misincorporation arising from this abundant adduct can be considered as potentially mutagenic as a highly miscoding but rare lesion.
Asunto(s)
Aductos de ADN/metabolismo , Replicación del ADN , ADN Polimerasa Dirigida por ADN/metabolismo , Desoxiguanosina/análogos & derivados , Daño del ADN , Reparación del ADN , ADN Polimerasa Dirigida por ADN/genética , Desoxiguanosina/metabolismo , Humanos , Estructura MolecularRESUMEN
KatG is a bifunctional, heme-dependent enzyme in the front-line defense of numerous bacterial and fungal pathogens against H2O2-induced oxidative damage from host immune responses. Contrary to the expectation that catalase and peroxidase activities should be mutually antagonistic, peroxidatic electron donors (PxEDs) enhance KatG catalase activity. Here, we establish the mechanism of synergistic cooperation between these activities. We show that at low pH values KatG can fully convert H2O2 to O2 and H2O only if a PxED is present in the reaction mixture. Stopped-flow spectroscopy results indicated rapid initial rates of H2O2 disproportionation slowing concomitantly with the accumulation of ferryl-like heme states. These states very slowly returned to resting (i.e. ferric) enzyme, indicating that they represented catalase-inactive intermediates. We also show that an active-site tryptophan, Trp-321, participates in off-pathway electron transfer. A W321F variant in which the proximal tryptophan was replaced with a non-oxidizable phenylalanine exhibited higher catalase activity and less accumulation of off-pathway heme intermediates. Finally, rapid freeze-quench EPR experiments indicated that both WT and W321F KatG produce the same methionine-tyrosine-tryptophan (MYW) cofactor radical intermediate at the earliest reaction time points and that Trp-321 is the preferred site of off-catalase protein oxidation in the native enzyme. Of note, PxEDs did not affect the formation of the MYW cofactor radical but could reduce non-productive protein-based radical species that accumulate during reaction with H2O2 Our results suggest that catalase-inactive intermediates accumulate because of off-mechanism oxidation, primarily of Trp-321, and PxEDs stimulate KatG catalase activity by preventing the accumulation of inactive intermediates.
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Proteínas Bacterianas/metabolismo , Catalasa/metabolismo , Modelos Moleculares , Peroxidasa/metabolismo , Algoritmos , Sustitución de Aminoácidos , Proteínas Bacterianas/agonistas , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Benzotiazoles/farmacología , Biocatálisis/efectos de los fármacos , Catalasa/química , Catalasa/genética , Dominio Catalítico , Espectroscopía de Resonancia por Spin del Electrón , Transporte de Electrón/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Depuradores de Radicales Libres/farmacología , Peróxido de Hidrógeno/metabolismo , Concentración de Iones de Hidrógeno , Mutagénesis Sitio-Dirigida , Mutación , Oxidación-Reducción , Peroxidasa/química , Peroxidasa/genética , Conformación Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Ácidos Sulfónicos/farmacología , Triptófano/químicaRESUMEN
Catalase-peroxidases (KatGs), the only catalase-active members of their superfamily, all possess a 35-residue interhelical loop called large loop 2 (LL2). It is essential for catalase activity, but little is known about its contribution to KatG function. LL2 shows weak sequence conservation; however, its length is nearly identical across KatGs, and its apex invariably makes contact with the KatG-unique C-terminal domain. We used site-directed and deletion mutagenesis to interrogate the role of LL2 and its interaction with the C-terminal domain in KatG structure and catalysis. Single and double substitutions of the LL2 apex had little impact on the active site heme [by magnetic circular dichroism or electron paramagnetic resonance (EPR)] and activity (catalase or peroxidase). Conversely, deletion of a single amino acid from the LL2 apex reduced catalase activity by 80%. Deletion of two or more apex amino acids or all of LL2 diminished catalase activity by 300-fold. Peroxide-dependent but not electron donor-dependent kcat/KM values for deletion variant peroxidase activity were reduced 20-200-fold, and kon for cyanide binding diminished by 3 orders of magnitude. EPR spectra for deletion variants were all consistent with an increase in the level of pentacoordinate high-spin heme at the expense of hexacoordinate high-spin states. Together, these data suggest a shift in the distribution of active site waters, altering the reactivity of the ferric state, toward, among other things, compound I formation. These results identify the importance of LL2 length conservation for maintaining an intersubunit interaction that is essential for an active site water distribution that facilitates KatG catalytic activity.
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Proteínas Arqueales/química , Proteínas Bacterianas/química , Hemo/química , Peroxidasas/química , Pliegue de Proteína , Sustitución de Aminoácidos , Proteínas Arqueales/genética , Proteínas Bacterianas/genética , Dominio Catalítico , Dicroismo Circular , Espectroscopía de Resonancia por Spin del Electrón , Hemo/genética , Hemo/metabolismo , Mutagénesis Sitio-Dirigida , Mutación Missense , Peroxidasas/genética , Estructura Secundaria de ProteínaRESUMEN
Deinococcus spp are among the most radiation-resistant micro-organisms that have been discovered. They show remarkable resistance to a range of damage caused by ionizing radiation, desiccation, UV radiation and oxidizing agents. Traditionally, Escherichia coli and Saccharomyces cerevisiae have been the two platforms of choice for engineering micro-organisms for biotechnological applications, because they are well understood and easy to work with. However, in recent years, researchers have begun using Deinococcus spp in biotechnologies and bioremediation due to their specific ability to grow and express novel engineered functions. More recently, the sequencing of several Deinococcus spp and comparative genomic analysis have provided new insight into the potential of this genus. Features such as the accumulation of genes encoding cell cleaning systems that eliminate organic and inorganic cell toxic components are widespread among Deinococcus spp. Other features such as the ability to degrade and metabolize sugars and polymeric sugars make Deinococcus spp. an attractive alternative for use in industrial biotechnology.
Asunto(s)
Deinococcus/genética , Microbiología Industrial , Biopelículas , Biotecnología , Pared Celular/química , Deinococcus/citología , Deinococcus/fisiología , Microbiología Industrial/instrumentación , Microbiología Industrial/métodos , Estrés OxidativoRESUMEN
Catalase-peroxidase (KatG) is found in eubacteria, archaea, and lower eukaryotae. The enzyme from Mycobacterium tuberculosis has received the greatest attention because of its role in activation of the antitubercular pro-drug isoniazid, and the high frequency with which drug resistance stems from mutations to the katG gene. Generally, the catalase activity of KatGs is striking. It rivals that of typical catalases, enzymes with which KatGs share no structural similarity. Instead, catalatic turnover is accomplished with an active site that bears a strong resemblance to a typical peroxidase (e.g., cytochrome c peroxidase). Yet, KatG is the only member of its superfamily with such capability. It does so using two mutually dependent cofactors: a heme and an entirely unique Met-Tyr-Trp (MYW) covalent adduct. Heme is required to generate the MYW cofactor. The MYW cofactor allows KatG to leverage heme intermediates toward a unique mechanism for H2O2 oxidation. This review evaluates the range of intermediates identified and their connection to the diverse catalytic processes KatG facilitates, including mechanisms of isoniazid activation.
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Archaea/enzimología , Bacterias/enzimología , Catalasa/metabolismo , Coenzimas/metabolismo , Peroxidasa/metabolismo , Antituberculosos/metabolismo , Archaea/química , Bacterias/química , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Catalasa/química , Coenzimas/química , Hemo/química , Hemo/metabolismo , Isoniazida/metabolismo , Metionina/química , Metionina/metabolismo , Modelos Moleculares , Mycobacterium tuberculosis/química , Mycobacterium tuberculosis/enzimología , Peroxidasa/química , Profármacos/metabolismo , Triptófano/química , Triptófano/metabolismo , Tirosina/química , Tirosina/metabolismoRESUMEN
During spring 2012, potted impatiens (Impatiens walleriana Hook.f.) plants with symptoms of a foliar disease were found in several commercial greenhouses in Mobile County, Alabama. Symptomatic leaves were chlorotic with no distinct lesions, and quickly wilted and abscised from erect green stems. In summer 2012 and 2013, numerous landscape impatiens plants with similar symptoms were observed in a large area from Mobile County north to Lee County, Alabama. A downy mildew was observed on the lower surfaces of symptomatic and abscised leaves from all locations. It consisted of hyaline, monopodial sporangiophores and ovoid, hyaline sporangia. Sporangiophores, which emerged from stomata, consisted of apical branches arranged at right angles to the supporting branches; they measured 69 to 90 µm long with individual branches measuring 7 to 14 µm long. Sporangia were borne on the tips of sporangiophore branches and measured 10 to 16.5 × 17 to 22.5 µm. No oospores were observed. In 2013, symptomatic plants were obtained from two separate locations in Alabama (Mobile and Tallapoosa counties). Total genomic DNA was extracted directly from symptomatic plant tissue and the large ribosomal subunit DNA was amplified by PCR using primers NL-1 and NL-4 (1). From both isolates, amplicons of 600 and 775 bp were obtained. DNA from each amplicon of both isolates was purified, sequenced, and the sequences were deposited in GenBank (Accession Nos. KF956518 to 21). The sequences of the 600-bp amplicons were 99% similar to that of I. walleriana (JX142135); the sequences of the 775-bp amplicons were 99% similar to Plasmopara obducens isolates from Florida (JX217746), Ohio (JX142134), Serbia (HQ246451), and the United Kingdom (AY587558). In pathogenicity tests, 10 potted impatiens plants, I. walleriana'Super Elfin,' were inoculated with a sporangial suspension (1 × 105 sporangia/ml washed from infected leaves) from the Mobile County isolate, by spraying until runoff. Controls were inoculated with sterile water. Plants were incubated in a moist chamber at 21°C for 48 h and then maintained in a greenhouse at 22 to 25°C until symptom development. All inoculated plants developed symptoms of downy mildew within 10 days. Microscopic examination of the symptomatic tissue revealed sporangiophores and sporangia similar to those observed in naturally infected plants. Control plants showed no symptoms. To our knowledge, this is the first report of downy mildew caused by P. obducens on impatiens in Alabama. This disease has been reported to have a significant economic impact for growers throughout the United States (2,3). Impatiens downy mildew is likely to be a recurring problem in Alabama. References: (1) K. O'Donnell. Curr. Genet. 22:213, 1992. (2) A. Palmateer et al. Plant Dis. 97:687, 2013. (3) S. Wegulo et al. Plant Dis. 88:909, 2004.
RESUMEN
Bacterial gall symptoms were observed on Loropetalum chinense (R. Br.) Oliv. in two separate commercial nurseries in South Alabama during the spring of 2012. Limb dieback and plant death was first reported by the growers. Plants with dieback symptoms had galling and irregular dark callus formation on the lower stem and lower branches. Galls were small, 0.2 to 1 cm, inconspicuous, and in some cases girdled the stem causing breakage of the main stem. In both locations, 30 to 40% of the crop was affected. Similar symptoms have been observed on L. chinense in nursery and landscape plantings in central Alabama, North Carolina, and Georgia in previous years. Bacterial colonies were isolated from four plants representing two different locations. Isolates were recovered from surface sterilized symptomatic tissue on nutrient agar and King's medium B (KMB). All isolates were gram-negative and fluoresced blue-green under UV light after 48 h of growth at 28°C on KMB. One representative isolate from each site was identified as Pseudomonas savastanoi based on their fatty acid profiles (similarity index of 0.776; MIS-TSBA, version 4.0, MIDI Inc., Newark, DE) and LOPAT tests (2). The identity was confirmed by sequencing a 900-bp portion of the 16S rDNA gene, which revealed 98% similarity to the P. savastanoi type strain in NCBI (Accession No. AB021402). In greenhouse pathogenicity tests, eight Loropetalum liners were inoculated with a bacterial suspension (107 CFU/ml) of each of the two isolates. Plants were inoculated by injecting the suspension into the lower stem after wounding by puncturing with needles or slicing sections of the bark. Controls were inoculated with water. All plants inoculated with the bacteria developed gall symptoms in 8 weeks under 90% relative humidity at 30°C. The bacteria were reisolated from five inoculated plants. DNA was extracted from each isolate, amplified using primer pair 27F/1492R targeting the 16S rDNA gene (1), and sequenced. Sequences (900 bp) from all isolates shared 98 to 99% similarity to P. savastanoi type strain in GenBank (Accession No. AB021402). Nucleotide sequence data reported are available in GenBank under accessions JX915832 to 37. To our knowledge, this is the first report of bacterial gall of L. chinense caused by P. savastanoi in the United States. Given the increasing prevalence of this disease in South Alabama, its confirmation is a significant step toward management recommendations for growers. References: (1) D. J. Lane. 16S/23S rRNA sequencing. Page 115-175 in: Nucleic Acid Techniques in Bacterial Systematics. E. Stackebrandt and M. Goodfellow, eds. John Wiley and Sons, New York, 1991. (2) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2001.
RESUMEN
Bovine liver supplements are sought after by consumers due to their nutrient-dense profile and high protein content. However, there is a lack of information regarding bovine liver supplement labeling practices. The objective of this study was to assess labeling practices and compliance with U.S. regulatory standards among commercially sold bovine liver supplements. The product labels for 49 bovine liver supplements were examined for required information, including a statement of identity; net quantity of contents; "Supplement Facts" label; ingredient statement; and name and place of business of manufacturer, packer, or distributor with domestic address or telephone number. Any claims made on the supplement labels were also reviewed for compliance. Overall, 59% of the products had at least one instance of noncompliance with U.S. labeling regulations. The main categories of noncompliance were missing a domestic mailing address or phone number (39% of products), noncompliant nutrient content claim (31% of products), and missing/noncompliant disclaimer for a nutritional support statement (6% of products). The lack of a mailing address or phone number is problematic because it prevents consumers from being able to report serious adverse events to the manufacturer. The majority (85%) of the nutrient content observed on product labels was found to be noncompliant. Noncompliant nutrient content claims are a major concern for consumers who rely on these claims to assess the nutritional benefits of a product. Overall, the results of this study revealed a lack of labeling compliance in bovine liver supplements, indicating a need for increased awareness and monitoring.
Asunto(s)
Suplementos Dietéticos , Etiquetado de Productos , Animales , Bovinos , HígadoRESUMEN
Dietary supplements containing bovine (subfamily Bovinae) liver are susceptible to fraud due to their high value and the lack of modern detection methods available for processed animal tissues. The objective of this research was to use molecular methods to authenticate dietary supplements claiming to contain bovine liver or beef liver through the verification of animal species and tissue type. A total of 53 bovine/beef liver dietary supplements were purchased from online sources. The presence of liver was verified with reverse transcription and real-time PCR testing for microRNA-122 (miR-122), which is highly expressed in liver tissue. Multiplex real-time PCR targeting domestic cattle (Bos taurus), horse (Equus caballus), sheep (Ovis aries), and pork (Sus scrofa) was used to verify species. Samples that failed species identification with multiplex real-time PCR underwent DNA mini-barcoding. Overall, bovine species were detected in 48/53 liver supplements: 35 samples were confirmed as domestic cattle with multiplex real-time PCR and an additional 13 samples were confirmed as domestic cattle or Bos spp. with DNA mini-barcoding. One of these samples was also positive for sheep/lamb, which was declared on the label. One product contained undeclared pork in addition to beef. MiR-122 was detected in 51 out of 53 supplements, suggesting the presence of liver. While this study demonstrates the potential use of tissue-specific microRNAs in verifying tissues in dietary supplements, more research is needed to evaluate the specificity of these markers.
Asunto(s)
ADN , MicroARNs , Animales , Bovinos , Suplementos Dietéticos , Caballos , Hígado , MicroARNs/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Ovinos , Especificidad de la EspecieRESUMEN
Gamma-ray bursts (GRBs) fall into two classes: short-hard and long-soft bursts. The latter are now known to have X-ray and optical afterglows, to occur at cosmological distances in star-forming galaxies, and to be associated with the explosion of massive stars. In contrast, the distance scale, the energy scale and the progenitors of the short bursts have remained a mystery. Here we report the discovery of a short-hard burst whose accurate localization has led to follow-up observations that have identified the X-ray afterglow and (for the first time) the optical afterglow of a short-hard burst; this in turn led to the identification of the host galaxy of the burst as a late-type galaxy at z = 0.16 (ref. 10). These results show that at least some short-hard bursts occur at cosmological distances in the outskirts of galaxies, and are likely to be caused by the merging of compact binaries.
RESUMEN
STUDY DESIGN: Cross-sectional comparison, control group. OBJECTIVES: To investigate the relationship between carotid arterial stiffness and circulating markers for cardiovascular disease (CVD) in spinal cord-injured (SCI) subjects compared with able-bodied (AB) individuals. SETTING: University Research Laboratory, University of Louisville. METHODS: SCI (n=14) and AB (n=13) subjects between 20-52 years of age were recruited to participate in the study. B-mode Doppler ultrasound was used to obtain carotid artery diameter measurements. Arterial stiffness was assessed via the stiffness index and distensibility coefficient. Markers of CVD risk were obtained by fasting blood draw. RESULTS: Carotid arterial stiffness index (P=0.061) and distensibility coefficient (P=0.370) were not different between the SCI and AB groups. The SCI group had higher high-sensitivity C-reactive protein (hsCRP) (P=0.046), triglycerides (P=0.017), leptin (P=0.040) and visfatin (P<0.001) compared with the control group. Visfatin (r=0.559, P=0.047), hsCRP (r=0.633, P=0.037), insulin (r=0.637, P=0.019) and HOMA (r=0.614, P=0.026) significantly correlated with carotid arterial stiffness index in the SCI group. CONCLUSION: This study demonstrated that SCI subjects are at a high cardiovascular risk as indicated by elevated hsCRP levels. Elevations in hsCRP and visfatin may contribute to accelerated atherogenic processes in the SCI population.
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Proteína C-Reactiva/metabolismo , Estenosis Carotídea/sangre , Estenosis Carotídea/fisiopatología , Citocinas/sangre , Nicotinamida Fosforribosiltransferasa/sangre , Traumatismos de la Médula Espinal/sangre , Traumatismos de la Médula Espinal/fisiopatología , Regulación hacia Arriba/fisiología , Rigidez Vascular/fisiología , Adulto , Aterosclerosis/sangre , Aterosclerosis/epidemiología , Aterosclerosis/fisiopatología , Biomarcadores/sangre , Proteína C-Reactiva/fisiología , Estenosis Carotídea/epidemiología , Comorbilidad , Estudios Transversales , Citocinas/fisiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Nicotinamida Fosforribosiltransferasa/fisiología , Traumatismos de la Médula Espinal/epidemiología , Adulto JovenRESUMEN
To understand whether fusions of thylakoid membranes from the parental chloroplasts occurred during zygote formation in Chlamydomonas reinhardtii, we performed an ultrastructural analysis of the zygotes produced by crossing mutants lacking photosystem I or II protein complexes, in the absence of de novo chloroplast protein synthesis. Thylakoid membranes from each parent could be distinguished on thin sections due to their organization in "supergrana" in mutants lacking photosystem I centers, by freeze-fracturing due to the absence of most of the exoplasmic-face (EF) particles in mutants lacking photosystem II centers, by immunocytochemistry using antibodies directed against photosystem II subunits. We demonstrate that a fusion of the thylakoid membranes occurred during zygote formation approximately 15 h after mating. These fusions allowed a lateral redistribution of the thylakoid membrane proteins. These observations provide the structural basis for the restoration of photosynthetic electron flow in the mature zygote that we observed in fluorescence induction experiments.
Asunto(s)
Chlamydomonas/ultraestructura , Cloroplastos/ultraestructura , Técnica de Fractura por Congelación , Prueba de Complementación Genética , Membranas Intracelulares/química , Membranas Intracelulares/ultraestructura , Fusión de Membrana , Proteínas de la Membrana/análisis , Microscopía Electrónica , Fotosíntesis , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Complejo de Proteína del Fotosistema I , Complejo de Proteína del Fotosistema II , Cigoto/ultraestructuraRESUMEN
We investigated the ultrastructure of thylakoid membranes that lacked either some or all of their Photosystem II centers in the F34SU3 and F34 mutants of Chlamydomonas reinhardtii. We obtained the following results: (a) There are no particles of the 160-A size class on the EF faces of the thylakoids in the absence of Photosystem II centers (as in F34); the F34SU3 contains 50% of the wild-type number of PSII centers and EF particles. (b) The density of the particles on the PF faces of the thylakoids is higher in the mutants than in the wild type. (c) The fluorescence analysis shows that the organization of the pigments is the same regardless of whether 50% of the PSII centers are temporarily inactivated (by preilluminating the wild type) or are actually missing from the thylakoid membrane (F34SU3). Our results, therefore, support a model in which: (a) each 160-A EF particle has only one PSII center surrounded by light-harvesting complexes and (b) part of the PSH antenna is associated with 80-A PF particles in both of the mutants and the wild type.
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Chlamydomonas/ultraestructura , Cloroplastos/ultraestructura , Fotosíntesis , Chlamydomonas/genética , Clorofila/análisis , Cloroplastos/análisis , Modelos Biológicos , Mutación , Péptidos/análisisRESUMEN
We studied the assembly of photosystem II (PSII) in several mutants from Chlamydomonas reinhardtii which were unable to synthesize either one PSII core subunit (P6 [43 kD], D1, or D2) or one oxygen-evolving enhancer (OEE1 or OEE2) subunit. Synthesis of the PSII subunits was analyzed on electrophoretograms of cells pulse labeled with [14C]acetate. Their accumulation in thylakoid membranes was studied on immunoblots, their chlorophyll-binding ability on nondenaturating gels, their assembly by detergent fractionation, their stability by pulse-chase experiments and determination of in vitro protease sensitivity, and their localization by immunocytochemistry. In Chlamydomonas, the PSII core subunits P5 (47 kD), D1, and D2 are synthesized in a concerted manner while P6 synthesis is independent. P5 and P6 accumulate independently of each other in the stacked membranes. They bind chlorophyll soon after, or concomitantly with, their synthesis and independently of the presence of the other PSII subunits. Resistance to degradation increases step by step: beginning with assembly of P5, D1, and D2, then with binding of P6, and, finally, with binding of the OEE subunits on two independent high affinity sites (one for OEE1 and another for OEE2 to which OEE3 binds). In the absence of PSII cores, the OEE subunits accumulate independently in the thylakoid lumen and bind loosely to the membranes; OEE1 was found on stacked membranes, but OEE2 was found on either stacked or unstacked membranes depending on whether or not P6 was synthesized.
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Chlamydomonas/genética , Clorofila/genética , Mutación , Fotosíntesis , Proteínas de Plantas/genética , Procesamiento Proteico-Postraduccional , Chlamydomonas/metabolismo , Chlamydomonas/ultraestructura , Clorofila/biosíntesis , Clorofila/aislamiento & purificación , Clorofila/metabolismo , Membranas Intracelulares/metabolismo , Membranas Intracelulares/ultraestructura , Cinética , Complejos de Proteína Captadores de Luz , Sustancias Macromoleculares , Péptido Hidrolasas/metabolismo , Proteínas del Complejo del Centro de Reacción Fotosintética , Complejo de Proteína del Fotosistema II , Proteínas de Plantas/biosíntesis , Proteínas de Plantas/aislamiento & purificaciónRESUMEN
REASONS FOR PERFORMING STUDY: Marginal osteophytes represent a well known component of osteoarthritis in man and animals. Conversely, central subchondral osteophytes (COs), which are commonly present in human knees with osteoarthritis, have not been reported in horses. OBJECTIVES: To describe and compare computed radiography (CR), single-slice computed tomography (CT), 1.5 Tesla magnetic resonance imaging (MRI), and histological features of COs in equine metacarpophalangeal joints with macroscopic evidence of naturally-occurring osteoarthritis. METHODS: MRI sequences (sagittal spoiled gradient recalled echo [SPGR] with fat saturation, sagittal T2-weighted fast spin echo with fat saturation [T2-FS], dorsal and transverse T1-weighted gradient-recalled echo [GRE], and sagittal T2*-weighted gradient echo with fast imaging employing steady state acquisition [FIESTA]), as well as transverse and reformatted sagittal CTI and 4 computed radiographic (CR) views of 20 paired metacarpophalangeal joints were acquired ex vivo. Following macroscopic evaluation, samples were harvested in predetermined sites of the metacarpal condyle for subsequent histology. The prevalence and detection level of COs was determined for each imaging modality. RESULTS: Abnormalities consistent with COs were clearly depicted on MRI, using the SPGR sequence, in 7/20 (35%) joints. They were identified as a focal hypointense protuberance from the subchondral plate into the cartilage, at the palmarodistal aspect (n=7) and/or at the very dorsal aspect (n=2) of the metacarpal condyle. COs were visible but less obvious in 5 of the 7 joints using FIESTA and reformatted sagittal CT, and were not identifiable on T2-FS, T1-GRE or CR. Microscopically, they consisted of dense bone protruding into the calcified cartilage and disrupting the tidemarks, and they were consistently associated with overlying cartilage defects. CONCLUSIONS: Subchondral osteophytes are a feature of osteoarthritis of equine metacarpophalangeal joints and they may be diagnosed using 1.5 Tesla MRI and CT. POTENTIAL RELEVANCE: Central subchondral osteophytes on MRI represent indirect evidence of cartilage damage in horses.
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Miembro Anterior/patología , Enfermedades de los Caballos/patología , Imagen por Resonancia Magnética/veterinaria , Osteoartritis/veterinaria , Osteofito/veterinaria , Animales , Cadáver , Caballos , Osteoartritis/patología , Osteofito/patologíaRESUMEN
In a grounded theory study examining the process of precepting an unsafe student, it was found that preceptors assigned passing grades to students who in fact should not have passed. Although preceptors perceived their role as gatekeepers for the profession, by not assigning failing grades to students who should not have passed a course, essentially they were abdicating their responsibility. Indeed, the simple act of assuming responsibility for precepting a student implies professional as well as pedagogical accountability.
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Educación en Enfermería/normas , Escolaridad , Preceptoría/normasRESUMEN
BACKGROUND: Extensive osteochondritis dissecans (OCD) lesions of the lateral ridge of the trochlea of the femur (LRTF), the most common OCD-affected site in the stifle, have a poor outcome with surgical debridement and can be career ending. The early detection of osteochondrosis lesions and their conservative management holds the promise to enhance outcome. We hypothesise that ultrasonography is a valuable field screening tool to detect and monitor early subclinical LRTF osteochondrosis. OBJECTIVES: The goals were to 1) describe the normal ultrasonographic features of the LRTF in foals of different ages and 2) screen a foal cohort at the farm for early subclinical osteochondrosis lesions. STUDY DESIGN: Prospective cohort study. METHODS: The LRTF of both hindlimbs of Thoroughbred foals (n = 46, 27-166 days old) were imaged once with ultrasonography and radiography (lateromedial and caudolateral-craniomedial oblique views). Cartilage thickness, ossification front indentation of the chondro-osseous junction and epiphyseal vascularisation were assessed on ultrasonography. Follow-up radiographs were taken as yearlings. RESULTS: The cartilage thickness, ossification front indentation and epiphyseal vascularisation significantly decreased with advancing maturity. Subclinical osteochondrosis lesions, characterised by semicircular indentations in the ossification front (indirect evidence of focal failure of ossification and retained cartilage) were detected in six foals (28-145 days old), both with radiography and ultrasonography. Ultrasonography provided a better overall subjective assessment of the osteochondrosis lesion topography (length, depth and the width) compared with radiography. MAIN LIMITATIONS: Post-mortem validation of lesions was not possible. CONCLUSION: Ultrasonography of the LRTF is a practical, inexpensive and reliable technique to discriminate physiological from pathological events at the LRTF in young foals. It revealed the complex topography of the chondro-osseous junction permitting a rapid, comprehensive assessment of the subclinical osteochondrosis lesions in very young foals.
Asunto(s)
Miembro Posterior/diagnóstico por imagen , Enfermedades de los Caballos/diagnóstico por imagen , Osteocondrosis/veterinaria , Envejecimiento , Animales , Cartílago Articular/irrigación sanguínea , Cartílago Articular/patología , Epífisis/irrigación sanguínea , Epífisis/diagnóstico por imagen , Femenino , Caballos , Masculino , Osteocondrosis/diagnóstico por imagen , Estudios ProspectivosRESUMEN
KEY HIGHLIGHTS: 1. Measles eradication is the ultimate goal but it is premature to set a date for its accomplishment. Existing regional elimination goals should be vigorously pursued to enable setting a global target by 2020. 2. The basic strategic approaches articulated in the Global Measles and Rubella Strategic Plan 2012-2020 are valid to achieve the goals but have not been fully implemented (or not appropriately adapted to local situations). 3. The report recommends a shift from primary reliance on supplementary immunization activities (SIAs) to assure two doses of measles-containing vaccine (MCV) are delivered to the target population to primary reliance on ongoing services to assure administration of two doses of MCV. Regular high quality SIAs will still be necessary while ongoing services are being strengthened. 4. The report recommends a shift from primary reliance on coverage to measure progress to incorporating disease incidence as a major indicator. 5. The report recommends that the measles/rubella vaccination program be considered an indicator for the quality of the overall immunization program and that measles/rubella incidence and measles and rubella vaccination coverage be considered as primary indicators of immunization program performance. 6. Polio transition presents both risks and opportunities: risks should be minimized and opportunities maximized. 7. A school entry immunization check could contribute significantly to strengthening overall immunization services with assurance that recommended doses of measles and rubella vaccines as well as other vaccines have been delivered and providing those vaccines at that time if the child is un- or under-vaccinated. 8. Program decisions should increasingly be based on good quality data and appropriate analysis. 9. The incorporation of rubella vaccination into the immunization program needs to be accelerated - it should be accorded equivalent emphasis as measles. 10. Outbreak investigation and response are critical but the most important thing is to prevent outbreaks.
Asunto(s)
Salud Global , Planificación en Salud , Programas de Inmunización , Sarampión/prevención & control , Rubéola (Sarampión Alemán)/prevención & control , Erradicación de la Enfermedad , Salud Global/historia , Planificación en Salud/historia , Planificación en Salud/métodos , Historia del Siglo XXI , Humanos , Programas de Inmunización/historia , Vacuna Antisarampión/administración & dosificación , Vacuna Antisarampión/inmunología , Prevalencia , Vacuna contra la Rubéola/administración & dosificación , Vacuna contra la Rubéola/inmunologíaRESUMEN
BACKGROUND: Noninvasive imaging tools are needed to screen foal femoropatellar joints to detect subclinical osteochondrosis lesions due to focal failure of endochondral ossification to enhance early management to optimise intrinsic healing events. Recently investigations employing 3T susceptibility-weighted magnetic resonance imaging (3T SWI MRI) and CT have demonstrated their capacity for early osteochondrosis diagnosis, but these technologies are not practical for field screening. We postulate that ultrasonography is a valuable field tool for the detection of subclinical osteochondrosis lesions. OBJECTIVES: The goals were to 1) describe the ultrasonographic features of the femoral trochlea of healthy and osteochondrosis-predisposed neonatal foals, 2) validate the capacity of ultrasound to assess cartilage canal vascular archictecture and the ossification front and 3) evaluate field feasibility in a pilot study. STUDY DESIGN: Experimental study. METHODS: Ultrasonographic evaluation of osteochondrosis predisposed (n = 10) and control (n = 6) femoral trochleas was performed ex vivo and compared with site-matched histological sections and 3T SWI MRI. The articular and epiphyseal cartilage thickness, ossification front indentation and cartilage canal vascular archictecture were assessed at each ROI. Femoral trochleae of foals (n = 3) aged ≈ 1, 3 and 6 months were also evaluated with ultrasonography in field. RESULTS: Ultrasonographic measurements strongly correlated with the histological measurements. There was no difference in the cartilage thickness or ossification front indentation between control and osteochondrosis-predisposed specimens. The cartilage canal vascular archictecture on ultrasonograms corresponded with the vessel pattern observed on site matched histology and 3T SWI MRI. MAIN LIMITATIONS: The number of specimens for study was limited and no early osteochondrosis lesions were present within the predilected group, but a field study is now underway. CONCLUSION: Ultrasonographic examination of the femoral trochlea permitted accurate evaluation of cartilage thickness, cartilage canal vascular archictecture and ossification front indentation in young foals and is a promising, practical tool for screening subclinical osteochondrosis and monitoring and managing lesions at important clinical sites.
Asunto(s)
Fémur/diagnóstico por imagen , Placa de Crecimiento/diagnóstico por imagen , Enfermedades de los Caballos/diagnóstico por imagen , Osteocondrosis/veterinaria , Animales , Animales Recién Nacidos , Femenino , Caballos , Imagen por Resonancia Magnética/veterinaria , Masculino , Osteocondrosis/diagnóstico por imagen , Ultrasonografía/normas , Ultrasonografía/veterinariaRESUMEN
Tolstoy reports the existence of a characteristic 100 thousand year (ky) period in the bathymetry of fast-spreading seafloor but does not argue that sea level change is a first-order control on seafloor morphology worldwide. Upon evaluating the overlap between tectonic and Milankovitch periodicities across spreading rates, we reemphasize that fast-spreading ridges are the best potential recorders of a sea level signature in seafloor bathymetry.