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1.
Foods ; 13(5)2024 Feb 29.
Artículo en Inglés | MEDLINE | ID: mdl-38472873

RESUMEN

Maintaining maize quality while drying during a rainy season is a major challenge for smallholder farmers in developing countries. We conducted a study to evaluate the impact of temporarily storing wet maize of 18, 21, and 24% moisture content (m.c.) in hermetic Purdue Improved Crop Storage (PICS) and polypropylene (PP) woven (control) bags for 21 days. Oxygen and carbon dioxide concentrations were monitored, and m.c., germination, and visual mold were assessed. In PICS bags, oxygen dropped below 1% within 7, 11.5, and 21 days for maize at 24, 21, and 18% m.c., respectively. After 21 days, the m.c. of maize stored in PICS bags remained constant, but decreased in PP bags. Germination of maize in PICS bags decreased by 0.5, 6.2, and 95.5 percentage points for 18, 21, and 24% m.c., respectively. In PP bags, germination decreased by 17.5, 15.2, and 39.5 percentage points for the respective moisture levels. After 21 days of storage, visible mold was present on maize stored in PP bags at both 21 and 24% m.c. No mold was observed on maize stored in PICS bags, but a fermentation smell was released from maize at 21 and 24% m.c. The results indicate that maize can be effectively stored in PICS bags at 21% m.c. or below for 21 days with minimal germination loss or mold growth. These findings highlight the potential of using hermetic bags for short-term grain quality preservation just before and during drying. This new utility adds to the current use of hermetic bags for protection against pests during long-term storage. Hermetic bags' dual functionality could significantly improve postharvest management on smallholder farms, thereby enhancing food and nutritional security and safety. Field testing is required in order to integrate this approach under smallholder farmers' conditions (e.g., temperature, m.c., drying practices, etc.).

2.
Foods ; 12(4)2023 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-36832966

RESUMEN

Maize is a major crop grown in many regions of the world for human consumption, starch production, and animal feed. After harvest, maize is dried to avoid spoilage caused by fungal growth. However, in the humid tropics, drying maize harvested during the rainy season poses challenges. In such instances, temporary storing maize under hermetic conditions may preserve grain quality while waiting for conditions suitable for drying. Wet maize at the moisture contents (m.c.) of 18, 21, and 24% was stored for up to 21 days in both hermetic and non-hermetic jars. The stored maize was assessed, every 7 days, for germination and related parameters, presence of visible mold, and pH. After 21 days of storage at 18, 21, and 24% m.c., maize germination decreased by 28.5, 25.2, and 95.5 percentage points, respectively, in hermetic jars; and by 28.5, 25.2, and 94.5 percentage points in non-hermetic jars (control). There was visible mold on maize stored in non-hermetic jars after 21 days regardless of m.c. Maize at 21 and 24% m.c. stored in hermetic conditions underwent lactic acid fermentation that reduced the pH. The findings suggest that maize at 18 and 21% m.c. can be stored for 14 and 7 days, respectively, under hermetic conditions without significant loss of quality. Further research is needed to thoroughly assess the application of these findings for temporarily storing and subsequently drying maize on farms and along the grain value chain.

3.
J Pharm Sci ; 112(4): 1052-1063, 2023 04.
Artículo en Inglés | MEDLINE | ID: mdl-36481417

RESUMEN

Most food, pharmaceutical, and chemical industries rely heavily on the supply of free-flowing powders that finds their application in raw materials, additives, and manufactured products. Improper storage conditions combined with environmental factors affect the free-flowing ability of powders. An undesirable transformation of these free-flowing powders into a coherent mass that resists flow is called caking. An important metric that can be used to measure the caking propensity of a material is the tensile strength, which is essentially the resistant stress needed to separate two layers of materials using an isostatic tensile strain. Even though several models have quantified the propensity of caking, the complex nature of interactions between the powder and its micro-environment makes the prediction of caking a challenging task. In the present work, the onset of tensile strength in isomalt with changes in temperature, relative humidity, and consolidation pressures using a shear cell was modeled using a finite element approach. The study found that at a consolidation pressure of 3 kPa and relative humidity of 85±0.1%, an increase in temperature by 5˚C increased the tensile strength of isomalt by a factor of 2. On the other hand, at a constant temperature of 25˚C, an increase in relative humidity from 85±0.1% to 86±0.1% registered an increase in tensile strength by 42.7%. This study also found that an increase in consolidation pressure from 3 kPa to 6 and 9 kPa increased the tensile strength by a factor of 1.79 and 2.54, respectively. The model had good agreement with the measurements and had an overall MAPE of 12.13%. This model can be applied to study the influence of extrinsic parameters on the propensity of caking during storage of bulk solids.


Asunto(s)
Disacáridos , Resistencia a la Tracción , Polvos , Análisis de Elementos Finitos
4.
Phys Rev Lett ; 126(8): 082301, 2021 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-33709760

RESUMEN

Quasielastic ^{12}C(e,e^{'}p) scattering was measured at spacelike 4-momentum transfer squared Q^{2}=8, 9.4, 11.4, and 14.2 (GeV/c)^{2}, the highest ever achieved to date. Nuclear transparency for this reaction was extracted by comparing the measured yield to that expected from a plane-wave impulse approximation calculation without any final state interactions. The measured transparency was consistent with no Q^{2} dependence, up to proton momenta of 8.5 GeV/c, ruling out the quantum chromodynamics effect of color transparency at the measured Q^{2} scales in exclusive (e,e^{'}p) reactions. These results impose strict constraints on models of color transparency for protons.

5.
Phys Rev Lett ; 125(26): 262501, 2020 Dec 31.
Artículo en Inglés | MEDLINE | ID: mdl-33449750

RESUMEN

We measure ^{2}H(e,e^{'}p)n cross sections at 4-momentum transfers of Q^{2}=4.5±0.5 (GeV/c)^{2} over a range of neutron recoil momenta p_{r}, reaching up to ∼1.0 GeV/c. We obtain data at fixed neutron recoil angles θ_{nq}=35°, 45°, and 75° with respect to the 3-momentum transfer q[over →]. The new data agree well with previous data, which reached p_{r}∼500 MeV/c. At θ_{nq}=35° and 45°, final state interactions, meson exchange currents, and isobar currents are suppressed and the plane wave impulse approximation provides the dominant cross section contribution. We compare the new data to recent theoretical calculations, where we observe a significant discrepancy for recoil momenta p_{r}>700 MeV/c.

6.
Am J Trop Med Hyg ; 100(2): 351-356, 2019 02.
Artículo en Inglés | MEDLINE | ID: mdl-30734697

RESUMEN

Hookworm infection causes anemia, malnutrition, and growth delay, especially in children living in sub-Saharan Africa. The World Health Organization recommends periodic mass drug administration (MDA) of anthelminthics to school-age children (SAC) as a means of reducing morbidity. Recently, questions have been raised about the effectiveness of MDA as a global control strategy for hookworms and other soil-transmitted helminths (STHs). Genomic DNA was extracted from Necator americanus hookworm eggs isolated from SAC enrolled in a cross-sectional study of STH epidemiology and deworming response in Kintampo North Municipality, Ghana. A polymerase chain reaction (PCR) assay was then used to identify single-nucleotide polymorphisms (SNPs) associated with benzimidazole resistance within the N. americanus ß-tubulin gene. Both F167Y and F200Y resistance-associated SNPs were detected in hookworm samples from infected study subjects. Furthermore, the ratios of resistant to wild-type SNP at these two loci were increased in posttreatment samples from subjects who were not cured by albendazole, suggesting that deworming drug exposure may enrich resistance-associated mutations. A previously unreported association between F200Y and a third resistance-associated SNP, E198A, was identified by sequencing of F200Y amplicons. These data confirm that markers of benzimidazole resistance are circulating among hookworms in central Ghana, with unknown potential to impact the effectiveness and sustainability of chemotherapeutic approaches to disease transmission and control.


Asunto(s)
Resistencia a Medicamentos/genética , Proteínas del Helminto/genética , Necator americanus/genética , Polimorfismo de Nucleótido Simple , Tubulina (Proteína)/genética , Animales , Antihelmínticos/farmacología , Bencimidazoles/farmacología , Niño , Estudios Transversales , Femenino , Expresión Génica , Marcadores Genéticos , Ghana/epidemiología , Infecciones por Uncinaria/tratamiento farmacológico , Infecciones por Uncinaria/epidemiología , Infecciones por Uncinaria/parasitología , Humanos , Masculino , Administración Masiva de Medicamentos/métodos , Necator americanus/efectos de los fármacos , Necator americanus/crecimiento & desarrollo , Cigoto/química , Cigoto/metabolismo
7.
Gene Expr Patterns ; 31: 1-6, 2019 01.
Artículo en Inglés | MEDLINE | ID: mdl-30468770

RESUMEN

Protein kinase A (PKA), also known as cAMP dependent protein kinase, is an essential component of many signaling pathways, many of which regulate key developmental processes. Inactive PKA is a tetrameric holoenzyme, comprised of two catalytic (PRKAC), and two regulatory subunits. Upon cAMP binding, the catalytic subunits are released and thereby activated. There are multiple isoforms of PKA catalytic subunits, but their individual roles are not well understood. In order to begin studying their roles in zebrafish development, it is first necessary to identify the spatial and temporal expression profiles for each prkac subunit. Here we evaluate the expression profiles for the four zebrafish prkacs: prkacαa, αb, ßa, and ßb, at key developmental time points: 24, 48 and 72 h post fertilization. We show that zebrafish prkacs are expressed throughout the developing nervous system, each showing unique expression patterns. This body of work will inform future functional studies into the roles of PKA during development.


Asunto(s)
Proteínas Quinasas Dependientes de AMP Cíclico/genética , Regulación del Desarrollo de la Expresión Génica , Proteínas de Pez Cebra/genética , Animales , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Sistema Nervioso/embriología , Sistema Nervioso/metabolismo , Subunidades de Proteína/genética , Subunidades de Proteína/metabolismo , Pez Cebra , Proteínas de Pez Cebra/metabolismo
8.
J Mol Biol ; 430(18 Pt B): 3297-3310, 2018 09 14.
Artículo en Inglés | MEDLINE | ID: mdl-29969581

RESUMEN

Heterogeneity in small heat shock proteins (sHsps) spans multiple spatiotemporal regimes-from fast fluctuations of part of the protein, to conformational variability of tertiary structure, plasticity of the interfaces, and polydispersity of the inter-converting, and co-assembling oligomers. This heterogeneity and dynamic nature of sHsps has significantly hindered their structural characterization. Atomic coordinates are particularly lacking for vertebrate sHsps, where most available structures are of extensively truncated homomers. sHsps play important roles in maintaining protein levels in the cell and therefore in organismal health and disease. HspB2 and HspB3 are vertebrate sHsps that are found co-assembled in neuromuscular cells, and variants thereof are associated with disease. Here, we present the structure of human HspB2/B3, which crystallized as a hetero-tetramer in a 3:1 ratio. In the HspB2/B3 tetramer, the four α-crystallin domains (ACDs) assemble into a flattened tetrahedron which is pierced by two non-intersecting approximate dyads. Assembly is mediated by flexible "nuts and bolts" involving IXI/V motifs from terminal regions filling ACD pockets. Parts of the N-terminal region bind in an unfolded conformation into the anti-parallel shared ACD dimer grooves. Tracts of the terminal regions are not resolved, most likely due to their disorder in the crystal lattice. This first structure of a full-length human sHsp heteromer reveals the heterogeneous interactions of the terminal regions and suggests a plasticity that is important for the cytoprotective functions of sHsps.


Asunto(s)
Proteínas de Choque Térmico HSP27/química , Proteínas de Choque Térmico/química , Modelos Moleculares , Conformación Proteica , Multimerización de Proteína , Secuencia de Aminoácidos , Proteínas de Choque Térmico HSP27/genética , Proteínas de Choque Térmico HSP27/metabolismo , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Humanos , Espectroscopía de Resonancia Magnética , Unión Proteica , Dominios y Motivos de Interacción de Proteínas
9.
J Dairy Sci ; 101(8): 7013-7026, 2018 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-29778471

RESUMEN

Milk protein concentrate (MPC) powders are widely used as ingredients for food product formulations due to their nutritional profile and sensory attributes. Processing parameters, storage conditions, and composition influences the flow properties of MPC powders. This study investigated the bulk and shear flow properties of 70.3, 81.5, and 88.1% (wt/wt, protein content) MPC after storage for 12 wk at 25 and 40°C. Additionally, the morphological and functional changes of the MPC powders were investigated and correlated with flowability. After 12 wk of storage at 25°C, the basic flow energy values significantly increased from 510 to 930 mJ as the protein content increased from 70 to 90% (wt/wt). Flow rate index was significantly higher for samples with high protein content. Dynamic flow tests indicated that MPC powders with high protein content displayed higher permeability. Shear tests confirmed that the samples stored at 25°C were more flowable than samples stored at 40°C. Likewise, the higher-protein content samples showed poor shear flow behavior. The results indicated that MPC powders stored at 25°C had less cohesiveness and better flow characteristics than MPC powders stored at 40°C. Overall, the MPC powders had markedly different flow properties due to their difference in composition and morphology. This study delivers insights on the particle morphology and flow behavior of MPC powders.


Asunto(s)
Manipulación de Alimentos/métodos , Conservación de Alimentos/métodos , Proteínas de la Leche/análisis , Animales , Tecnología de Alimentos/métodos , Polvos , Temperatura
10.
Food Sci Nutr ; 5(3): 617-624, 2017 05.
Artículo en Inglés | MEDLINE | ID: mdl-28572949

RESUMEN

Postharvest losses (PHL) are incurred in the tomato value chain in Ghana and solar drying of tomato is a promising technology for reducing the loss. However, there are concerns on the usage, functionality and sensory appeal of the dried products to consumers, compounded with the lack of information and research on dried tomato processing in Ghana. A survey was carried out by administering semistructured questionnaires to 395 randomly selected and willing respondents in the Accra Metropolis. Information was obtained on the socioeconomic profile, consumption pattern, knowledge, and acceptance of tomato processing technologies and assessment of quality attributes important to consumers. Most consumers (74%) preferred tomato powder that is conveniently packaged to retain the characteristic intense taste and the flavor using Friedman's rank mean procedure. The study indicated that consumers were more concerned about good manufacturing practices during the production of solar-dried tomato (48.8%) rather than the quality attributes (8.6%). These findings indicate the need for safe solar drying procedures in order to increase consumer acceptability of solar-dried tomato products in Ghana.

11.
Neuron ; 91(5): 1124-1136, 2016 Sep 07.
Artículo en Inglés | MEDLINE | ID: mdl-27568518

RESUMEN

Hippocampal replays are episodes of sequential place cell activity during sharp-wave ripple oscillations (SWRs). Conflicting hypotheses implicate awake replay in learning from reward and in memory retrieval for decision making. Further, awake replays can be forward, in the same order as experienced, or reverse, in the opposite order. However, while the presence or absence of reward has been reported to modulate SWR rate, the effect of reward changes on replay, and on replay direction in particular, has not been examined. Here we report divergence in the response of forward and reverse replays to changing reward. While both classes of replays were observed at reward locations, only reverse replays increased their rate at increased reward or decreased their rate at decreased reward, while forward replays were unchanged. These data demonstrate a unique relationship between reverse replay and reward processing and point to a functional distinction between different directions of replay. VIDEO ABSTRACT.


Asunto(s)
Hipocampo/citología , Hipocampo/fisiología , Células de Lugar/fisiología , Recompensa , Animales , Masculino , Modelos Neurológicos , Ratas
12.
Prev Vet Med ; 127: 121-33, 2016 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-26972273

RESUMEN

Bovine respiratory disease (BRD) is the most important cause of clinical disease and death in feedlot cattle. Respiratory viral infections are key components in predisposing cattle to the development of this disease. To quantify the contribution of four viruses commonly associated with BRD, a case-control study was conducted nested within the National Bovine Respiratory Disease Initiative project population in Australian feedlot cattle. Effects of exposure to Bovine viral diarrhoea virus 1 (BVDV-1), Bovine herpesvirus 1 (BoHV-1), Bovine respiratory syncytial virus (BRSV) and Bovine parainfluenza virus 3 (BPIV-3), and to combinations of these viruses, were investigated. Based on weighted seroprevalences at induction (when animals were enrolled and initial samples collected), the percentages of the project population estimated to be seropositive were 24% for BoHV-1, 69% for BVDV-1, 89% for BRSV and 91% for BPIV-3. For each of the four viruses, seropositivity at induction was associated with reduced risk of BRD (OR: 0.6-0.9), and seroincrease from induction to second blood sampling (35-60 days after induction) was associated with increased risk of BRD (OR: 1.3-1.5). Compared to animals that were seropositive for all four viruses at induction, animals were at progressively increased risk with increasing number of viruses for which they were seronegative; those seronegative for all four viruses were at greatest risk (OR: 2.4). Animals that seroincreased for one or more viruses from induction to second blood sampling were at increased risk (OR: 1.4-2.1) of BRD compared to animals that did not seroincrease for any viruses. Collectively these results confirm that prior exposure to these viruses is protective while exposure at or after feedlot entry increases the risk of development of BRD in feedlots. However, the modest increases in risk associated with seroincrease for each virus separately, and the progressive increases in risk with multiple viral exposures highlights the importance of concurrent infections in the aetiology of the BRD complex. These findings indicate that, while efficacious vaccines could aid in the control of BRD, vaccination against one of these viruses would not have large effects on population BRD incidence but vaccination against multiple viruses would be expected to result in greater reductions in incidence. The findings also confirm the multifactorial nature of BRD development, and indicate that multifaceted approaches in addition to efficacious vaccines against viruses will be required for substantial reductions in BRD incidence.


Asunto(s)
Complejo Respiratorio Bovino/epidemiología , Virus/aislamiento & purificación , Animales , Australia/epidemiología , Complejo Respiratorio Bovino/virología , Estudios de Casos y Controles , Bovinos , Femenino , Masculino , Prevalencia , Estudios Seroepidemiológicos
13.
Prev Vet Med ; 126: 159-69, 2016 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-26907209

RESUMEN

Viruses play a key role in the complex aetiology of bovine respiratory disease (BRD). Bovine viral diarrhoea virus 1 (BVDV-1) is widespread in Australia and has been shown to contribute to BRD occurrence. As part of a prospective longitudinal study on BRD, effects of exposure to BVDV-1 on risk of BRD in Australian feedlot cattle were investigated. A total of 35,160 animals were enrolled at induction (when animals were identified and characteristics recorded), held in feedlot pens with other cattle (cohorts) and monitored for occurrence of BRD over the first 50days following induction. Biological samples collected from all animals were tested to determine which animals were persistently infected (PI) with BVDV-1. Data obtained from the Australian National Livestock Identification System database were used to determine which groups of animals that were together at the farm of origin and at 28days prior to induction (and were enrolled in the study) contained a PI animal and hence to identify animals that had probably been exposed to a PI animal prior to induction. Multi-level Bayesian logistic regression models were fitted to estimate the effects of exposure to BVDV-1 on the risk of occurrence of BRD. Although only a total of 85 study animals (0.24%) were identified as being PI with BVDV-1, BVDV-1 was detected on quantitative polymerase chain reaction in 59% of cohorts. The PI animals were at moderately increased risk of BRD (OR 1.9; 95% credible interval 1.0-3.2). Exposure to BVDV-1 in the cohort was also associated with a moderately increased risk of BRD (OR 1.7; 95% credible interval 1.1-2.5) regardless of whether or not a PI animal was identified within the cohort. Additional analyses indicated that a single quantitative real-time PCR test is useful for distinguishing PI animals from transiently infected animals. The results of the study suggest that removal of PI animals and/or vaccination, both before feedlot entry, would reduce the impact of BVDV-1 on BRD risk in cattle in Australian feedlots. Economic assessment of these strategies under Australian conditions is required.


Asunto(s)
Diarrea Mucosa Bovina Viral/epidemiología , Virus de la Diarrea Viral Bovina Tipo 1 , Alimentación Animal/virología , Crianza de Animales Domésticos , Animales , Anticuerpos Antivirales/sangre , Diarrea Mucosa Bovina Viral/diagnóstico , Diarrea Mucosa Bovina Viral/prevención & control , Diarrea Mucosa Bovina Viral/transmisión , Bovinos , Estudios de Cohortes , Virus de la Diarrea Viral Bovina Tipo 1/genética , Virus de la Diarrea Viral Bovina Tipo 1/aislamiento & purificación , Prevalencia , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Factores de Riesgo , Vacunas Virales/administración & dosificación
14.
J Sci Food Agric ; 96(2): 359-64, 2016 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-26095631

RESUMEN

Flow difficulties during handling, storage, and processing are common in cereal grain-based powder industries. The many studies that focus on the flow properties of powders can be classified as flow indicators, shear properties, and dynamic flow properties. The non-uniformity of physical and chemical characteristics of the individual particles that make up the bulk solid of cereal grain-based powders adds complexity to the characterization of flow behavior. Even so, knowledge of flow behavior is critical to the design of productive and cost-effective equipment for handling and processing of these powders. Because many factors influence flow, a single property/index value may not satisfactorily quantify the flow or no-flow of powders. For powders of biological origin, chemical composition and environmental factors such as temperature and relative humidity complicate flow characterization. This review focuses on the specific flow characteristics that directly affect powder flow during handling, processing, and storage.


Asunto(s)
Grano Comestible/química , Manipulación de Alimentos , Polvos/química , Tamaño de la Partícula , Propiedades de Superficie
15.
J Agric Saf Health ; 22(3): 187-197, 2016 Jul 27.
Artículo en Inglés | MEDLINE | ID: mdl-29140630

RESUMEN

Grain dust explosions have been occurring in the U.S. for the past twenty years. In the past ten years, there have been an average of ten explosions a year, resulting in nine fatalities and 93 injuries. In more than half of these cases, the ignition source remains unidentified. The effect of ambient humidity on the likelihood of a dust explosion has been discussed for many years. However, no investigation into a possible link between the two has been carried out. In this study, we analyzed local weather data and grain dust explosions during the period 2006 to 2014 to measure potential relationships between the two events. The 84 analyzed explosions do not show any trend with regard to prevailing temperatures, or relative or absolute humidity. In addition, the ignition source could not be identified in 54 of the incidents. The majority of grain dust explosion incidents occurred at grain elevator facilities, where the dust generation potential was high compared with grain processing industries.


Asunto(s)
Accidentes de Trabajo/prevención & control , Traumatismos por Explosión/prevención & control , Polvo , Grano Comestible , Explosiones , Humanos , National Institute for Occupational Safety and Health, U.S. , Administración de la Seguridad , Estados Unidos , Tiempo (Meteorología)
16.
PLoS One ; 10(7): e0133226, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26208319

RESUMEN

Wnt signaling controls various aspects of developmental and cell biology, as well as contributing to certain cancers. Expression of the human Rho family small GTPase Wrch/RhoU is regulated by Wnt signaling, and Wrch and its paralog Chp/RhoV are both implicated in oncogenic transformation and regulation of cytoskeletal dynamics. We performed developmental genetic analysis of the single Caenorhabditis elegans ortholog of Chp and Wrch, CHW-1. Using a transgenic assay of the distal tip cell migration, we found that wild-type CHW-1 is likely to be partially constitutively active and that we can alter ectopic CHW-1-dependent migration phenotypes with mutations predicted to increase or decrease intrinsic GTP hydrolysis rate. The vulval P7.p polarity decision balances multiple antagonistic Wnt signals, and also uses different types of Wnt signaling. Previously described cooperative Wnt receptors LIN-17/Frizzled and LIN-18/Ryk orient P7.p posteriorly, with LIN-17/Fz contributing approximately two-thirds of polarizing activity. CHW-1 deletion appears to equalize the contributions of these two receptors. We hypothesize that CHW-1 increases LIN-17/Fz activity at the expense of LIN-18/Ryk, thus making the contribution of these signals unequal. For P7.p to polarize correctly and form a proper vulva, LIN-17/Fz and LIN-18/Ryk antagonize other Wnt transmembrane systems VANG-1/VanGogh and CAM-1/Ror. Our genetic data suggest that LIN-17/Fz represses both VANG-1/VanGogh and CAM-1/Ror, while LIN-18/Ryk represses only VANG-1. These data expand our knowledge of a sophisticated signaling network to control P7.p polarity, and suggests that CHW-1 can alter ligand gradients or receptor priorities in the system.


Asunto(s)
Proteínas de Caenorhabditis elegans/metabolismo , Polaridad Celular , Proteínas Tirosina Quinasas Receptoras/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Transducción de Señal , Secuencia de Aminoácidos , Animales , Caenorhabditis elegans , Proteínas de Caenorhabditis elegans/química , Proteínas de Caenorhabditis elegans/genética , Proteínas de Unión al Calcio/química , Proteínas de Unión al Calcio/metabolismo , Humanos , Modelos Biológicos , Datos de Secuencia Molecular , Mutación , Fenotipo , Proteínas Tirosina Quinasas Receptoras/genética , Receptores Acoplados a Proteínas G/genética , Alineación de Secuencia , Vía de Señalización Wnt
17.
BMC Struct Biol ; 15: 12, 2015 Jul 11.
Artículo en Inglés | MEDLINE | ID: mdl-26163297

RESUMEN

BACKGROUND: Stationary phase survival proteins (Sps) were found in Firmicutes as having analogous domain compositions, and in some cases genome context, as the resuscitation promoting factors of Actinobacteria, but with a different putative peptidoglycan cleaving domain. RESULTS: The first structure of a Firmicute Sps protein YuiC from B. subtilis, is found to be a stripped down version of the cell-wall peptidoglycan hydrolase MltA. The YuiC structures are of a domain swapped dimer, although some monomer is also found in solution. The protein crystallised in the presence of pentasaccharide shows a 1,6-anhydrodisaccharide sugar product, indicating that YuiC cleaves the sugar backbone to form an anhydro product at least on lengthy incubation during crystallisation. CONCLUSIONS: The structural simplification of MltA in Sps proteins is analogous to that of the resuscitation promoting factor domains of Actinobacteria, which are stripped down versions of lysozyme and soluble lytic transglycosylase proteins.


Asunto(s)
Fosfatasa Ácida/química , Bacillus subtilis/metabolismo , Proteínas Bacterianas/química , Bacillus subtilis/química , Dominio Catalítico , Cristalografía por Rayos X , Glicosiltransferasas/química , Modelos Moleculares , Oligosacáridos/metabolismo , Estructura Secundaria de Proteína
18.
Virology ; 481: 95-106, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-25771497

RESUMEN

The West Nile virus strain Kunjin virus (WNVKUN) NS4A protein is a multifunctional protein involved in many aspects of the virus life-cycle and is a major component of the WNVKUN replication complex (RC). Previously we identified a conserved region in the C-terminus of NS4A regulating proteolytic processing and RC assembly, and now investigate key conserved residues in the N-terminus of NS4A and their contribution to WNVKUN replication. Mutation of P13 completely ablated replication, whereas, mutation of P48 and D49, near the first transmembrane helix, and G66 within the helix, showed variable defects in replication, virion secretion and membrane proliferation. Intriguingly, the P48 and G66 NS4A mutants resulted in specific proteasome depletion of NS4A that could in part be rescued with a proteasome inhibitor. Our results suggest that the N-terminus of NS4A contributes to correct folding and stability, essential for facilitating the essential roles of NS4A during replication.


Asunto(s)
Membrana Celular/virología , Proteínas no Estructurales Virales/química , Proteínas no Estructurales Virales/metabolismo , Replicación Viral , Fiebre del Nilo Occidental/virología , Virus del Nilo Occidental/metabolismo , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Línea Celular , Secuencia Conservada , Humanos , Datos de Secuencia Molecular , Pliegue de Proteína , Estructura Secundaria de Proteína , Proteínas no Estructurales Virales/genética , Virus del Nilo Occidental/química , Virus del Nilo Occidental/genética , Virus del Nilo Occidental/crecimiento & desarrollo
19.
Structure ; 22(12): 1831-1843, 2014 Dec 02.
Artículo en Inglés | MEDLINE | ID: mdl-25435325

RESUMEN

Protein AMPylation, the transfer of AMP from ATP to protein targets, has been recognized as a new mechanism of host-cell disruption by some bacterial effectors that typically contain a FIC-domain. Eukaryotic genomes also encode one FIC-domain protein,HYPE, which has remained poorly characterized.Here we describe the structure of human HYPE, solved by X-ray crystallography, representing the first structure of a eukaryotic FIC-domain protein. We demonstrate that HYPE forms stable dimers with structurally and functionally integrated FIC-domains and with TPR-motifs exposed for protein-protein interactions. As HYPE also uniquely possesses a transmembrane helix, dimerization is likely to affect its positioning and function in the membrane vicinity. The low rate of auto AMPylation of the wild-type HYPE could be due to autoinhibition, consistent with the mechanism proposed for a number of putative FIC AMPylators. Our findings also provide a basis to further consider possible alternative cofactors of HYPE and distinct modes of target-recognition.


Asunto(s)
Proteínas Portadoras/metabolismo , Proteínas de la Membrana/metabolismo , Modelos Moleculares , Cristalografía por Rayos X , Humanos , Nucleotidiltransferasas , Conformación Proteica , Estructura Terciaria de Proteína , Relación Estructura-Actividad
20.
Vaccine ; 32(23): 2682-7, 2014 May 13.
Artículo en Inglés | MEDLINE | ID: mdl-24709588

RESUMEN

Epsilon toxin (Etx) is a ß-pore-forming toxin produced by Clostridium perfringens toxinotypes B and D and plays a key role in the pathogenesis of enterotoxemia, a severe, often fatal disease of ruminants that causes significant economic losses to the farming industry worldwide. This study aimed to determine the potential of a site-directed mutant of Etx (Y30A-Y196A) to be exploited as a recombinant vaccine against enterotoxemia. Replacement of Y30 and Y196 with alanine generated a stable variant of Etx with significantly reduced cell binding and cytotoxic activities in MDCK.2 cells relative to wild type toxin (>430-fold increase in CT50) and Y30A-Y196A was inactive in mice after intraperitoneal administration of trypsin activated toxin at 1000× the expected LD50 dose of trypsin activated wild type toxin. Moreover, polyclonal antibody raised in rabbits against Y30A-Y196A provided protection against wild type toxin in an in vitro neutralisation assay. These data suggest that Y30A-Y196A mutant could form the basis of an improved recombinant vaccine against enterotoxemia.


Asunto(s)
Toxinas Bacterianas/inmunología , Vacunas Bacterianas/inmunología , Enterotoxemia/prevención & control , Animales , Perros , Femenino , Células de Riñón Canino Madin Darby , Ratones , Ratones Endogámicos BALB C , Mutagénesis Sitio-Dirigida , Pruebas de Neutralización , Estructura Terciaria de Proteína , Conejos , Proteínas Recombinantes/inmunología , Vacunas Sintéticas/inmunología
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