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1.
J Microsc ; 277(3): 140-153, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31070241

RESUMO

A time-lapse study of thermomechanical fatigue damage has been undertaken using three-dimensional X-ray computer tomography. Morphologies were extracted from tomography data and integrated with data from microscopy modalities at different resolution levels. This enables contextualization of some of the fine-scale properties which underpin the large-scale damage observed via tomography. Lateral views of crack development are presented, which show networks analogous to mud-cracks. Crack fronts which develop in the most porous regions within the sintered attachment layer travel across the boundary into the copper substrate. The propagation characteristics of these cracks within the substrate are analysed. Evidence is provided of heterogeneous densification within the sintered joint under power cycling, and this is shown to play a major role in driving the initiation and propagation of the cracks. Examination of the texture (differing levels of X-ray absorption) of virtual cross-sectional images reveals the origins of the nonuniformity of densification. Finally, cracks within the sintered joint are shown to have a negligible impact on the conduction pathway of the joint due to their aspect ratio and orientation with respect to the assembly. LAY DESCRIPTION: This paper concerns the use of three-dimensional (3D) X-ray tomography, a nondestructive technique, to perform cradle-to-grave studies of sintered nanosilver die-attachments under operation. Sintered nanosilver die-attachments have been proposed as a more reliable and environmentally friendly alternative to solder alloy joints for emerging power electronics module designs. However, their degradation mechanisms are not as well understood. This same sample-study is about observing how the fine-scale structure of a sintered attachment evolves and degrades over time. Using 3D tomography affords otherwise infeasible perspectives, such as virtual cross-sections in the lateral plane of the attachment. These perspectives provide qualitative information which elucidates the degradation mechanisms. They demonstrate, for example, that the structure of the sintered attachment densifies under operation, and a consequence of this is the formation of shrinkage cracks in the most porous regions, much like mud-cracks. Other imaging techniques (metallographic etching and scanning electron microscopy) have been used in correlation with 3D renderings of these cracks to analyse their propagation and reveal their relationship both with the internal structure of the sintered attachment itself, and the structure of the substrate to which it is joined. It is shown that the cracks develop within the sintered attachment layer and eventually cross over into the substrate. A comparison of two sintered attachments with contrasting bulk porosities allows the effect of initial bond quality on crack development to be examined.

2.
Neuropathol Appl Neurobiol ; 45(4): 410-420, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-29770465

RESUMO

AIMS: To evaluate the relationship between expression of myxovirus-resistance protein A (MxA) protein on muscle biopsies by immunohistochemistry and disease activity in juvenile dermatomyositis (JDM) patients. Also, another aim was to investigate whether the expression of MxA is related with myositis-specific autoantibodies (MSA) status in JDM patients. METHODS: 103 patients (median aged 6.3, interquartile range 0.5-15.9) enrolled in the Juvenile Dermatomyositis Cohort and Biomarker Study (JDCBS). Muscle biopsies were stained with MxA and scored. Clinical data at initial presentation were collected and autoantibodies were analysed. Multiple linear regression analysis was performed to estimate the association between MxA expression on muscle fibres and muscle disease activity, and MSA status. RESULTS: Expression of MxA protein on JDM samples was identified in 61.2%. There was a significant association between MxA scores and Childhood Myositis Assessment Scale (CMAS) (P = 0.002), and Manual Muscle Testing of Eight Muscles (MMT8) (P = 0.026). CMAS and MMT8 scores were significantly lower in the group of patients with strong MxA expression. MxA scores differed according to MSA subgroups (P = 0.002). Patients with positive nuclear matrix protein 2 autoantibodies had strong MxA expression, whereas anti-melanoma differentiation-associated gene 5 positive patients had no or weak MxA expression. CONCLUSIONS: This study reveals the significant association between level of MxA expression on muscle fibres and clinical measures of muscular disease activity in JDM patients and MSA status. This confirms type I interferonopathies in muscle fibres of JDM patients which could help with improving treatment outcome in JDM patients and underscoring the distinct pathophysiological pathways in different MSA status.


Assuntos
Dermatomiosite/metabolismo , Doenças Musculares/imunologia , Miosite/metabolismo , Proteínas de Resistência a Myxovirus/metabolismo , Adolescente , Autoanticorpos/metabolismo , Biomarcadores/análise , Criança , Pré-Escolar , Estudos de Coortes , Dermatomiosite/imunologia , Feminino , Humanos , Lactente , Masculino , Miosite/imunologia , Proteínas de Resistência a Myxovirus/imunologia
3.
Appl Environ Microbiol ; 80(16): 5001-11, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24907337

RESUMO

Thermoanaerobacterium saccharolyticum, a Gram-positive thermophilic anaerobic bacterium, grows robustly on insoluble hemicellulose, which requires a specialized suite of secreted and transmembrane proteins. We report here the characterization of proteins secreted by this organism. Cultures were grown on hemicellulose, glucose, xylose, starch, and xylan in pH-controlled bioreactors, and samples were analyzed via spotted microarrays and liquid chromatography-mass spectrometry. Key hydrolases and transporters employed by T. saccharolyticum for growth on hemicellulose were, for the most part, hitherto uncharacterized and existed in two clusters (Tsac_1445 through Tsac_1464 for xylan/xylose and Tsac_1344 through Tsac_1349 for starch). A phosphotransferase system subunit, Tsac_0032, also appeared to be exclusive to growth on glucose. Previously identified hydrolases that showed strong conditional expression changes included XynA (Tsac_1459), XynC (Tsac_0897), and a pullulanase, Apu (Tsac_1342). An omnipresent transcript and protein making up a large percentage of the overall secretome, Tsac_0361, was tentatively identified as the primary S-layer component in T. saccharolyticum, and deletion of the Tsac_0361 gene resulted in gross morphological changes to the cells. The view of hemicellulose degradation revealed here will be enabling for metabolic engineering efforts in biofuel-producing organisms that degrade cellulose well but lack the ability to catabolize C5 sugars.


Assuntos
Proteínas de Bactérias/metabolismo , Hidrolases/metabolismo , Polissacarídeos/metabolismo , Thermoanaerobacterium/enzimologia , Proteínas de Bactérias/genética , Biodegradação Ambiental , Hidrolases/genética , Transporte Proteico , Thermoanaerobacterium/genética , Thermoanaerobacterium/metabolismo
5.
J Environ Radioact ; 228: 106513, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33360017

RESUMO

While radioisotopes of noble gases are known to be indicators of underground nuclear explosions (UNE), McIntyre et al. (2017) was the first to report the presence of 39Ar in shallow soil gas in association with a decades old UNE. While this finding hinted at the potential application of 39Ar to be used as an indicator of a UNE, doing so would also require an understanding of the natural concentrations of 39Ar present in soil gas. Without knowing the expected range and variability of naturally occurring concentrations of 39Ar, it is difficult to determine what measured concentrations would be indicative of an elevated concentration. This paper presents results from 16 soil gas samples and three atmospheric air samples collected from various locations across the western United States. Shallow soil gas samples were collected into self-contained underwater breathing apparatus (SCUBA) tanks using a custom-built soil gas sampling system and then processed and analyzed for 39Ar. The measured concentrations of 39Ar varied from atmospheric air concentrations to about 3.5 times atmospheric air concentrations (58 mBq/m3). The results presented here represent the first measurements of natural background 39Ar concentrations in shallow soil gas. This data will be necessary if 39Ar is to be used as an indicator of UNE.


Assuntos
Argônio , Radiação de Fundo , Monitoramento de Radiação , Radioisótopos , Poluentes Radioativos do Solo/análise , Argônio/análise , Radioisótopos/análise , Solo
6.
Science ; 153(3739): 1006-7, 1966 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-4958354

RESUMO

Blood forms of human vivax malaria infected splenectomized night monkeys (Aotus trivirgatus). Anopheles albimanus mosquitoes transmitted the in fection from a monkey to two human volunteers; parasites and symptoms ap peared 11 days later. Blood forms of vivax malaria from each of the two humans infected other night monkeys.


Assuntos
Azatioprina , Reservatórios de Doenças , Haplorrinos , Plasmodium vivax , Animais , Anopheles , Humanos , Esplenectomia
7.
Science ; 285(5435): 1889-92, 1999 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-10489362

RESUMO

The (56)Fe/(54)Fe of Fe-bearing phases precipitated in sedimentary environments varies by 2.5 per mil (delta(56)Fe values of +0.9 to -1. 6 per mil). In contrast, the (56)Fe/(54)Fe of Fe-bearing phases in igneous rocks from Earth and the moon does not vary measurably (delta(56)Fe = 0.0 +/- 0.3 per mil). Experiments with dissimilatory Fe-reducing bacteria of the genus Shewanella algae grown on a ferrihydrite substrate indicate that the delta(56)Fe of ferrous Fe in solution is isotopically lighter than the ferrihydrite substrate by 1.3 per mil. Therefore, the range in delta(56)Fe values of sedimentary rocks may reflect biogenic fractionation, and the isotopic composition of Fe may be used to trace the distribution of microorganisms in modern and ancient Earth.


Assuntos
Isótopos de Ferro/análise , Ferro/análise , Planeta Terra , Bacilos Gram-Negativos Anaeróbios Facultativos/metabolismo , Espectrometria de Massas/métodos , Lua
8.
Oncogene ; 26(11): 1661-72, 2007 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-17001320

RESUMO

von Hippel-Lindau (VHL) disease is a dominantly inherited family cancer syndrome characterized by the development of retinal and central nervous system haemangioblastomas, renal cell carcinoma (RCC) and phaeochromocytoma. Specific germline VHL mutations may predispose to haemangioblastomas, RCC and phaeochromocytoma to a varying extent. Although dysregulation of the hypoxia-inducible transcription factor-2 and JunB have been linked to the development of RCC and phaeochromocytoma, respectively, the precise basis for genotype-phenotype correlations in VHL disease have not been defined. To gain insights into the pathogenesis of RCC in VHL disease we compared gene expression microarray profiles in a RCC cell line expressing a Type 1 or Type 2B mutant pVHL (RCC-associated) to those of a Type 2A or 2C mutant (not associated with RCC). We identified 19 differentially expressed novel VHL target genes linked to RCC development. Eight targets were studied in detail by quantitative real-time polymerase chain reaction (three downregulated and five upregulated by wild-type VHL) and for six genes the effect of VHL inactivation was mimicked by hypoxia (but hypoxic-induction of smooth muscle alpha-actin 2 was specific for a RCC cell line). The potential role of four RCC-associated VHL target genes was assessed in vitro. NB thymosin beta (TMSNB) and proteinase-activated receptor 2 (PAR2) (both downregulated by wt pVHL) increased cell growth and motility in a RCC cell line, but aldehyde dehydrogenase (ALDH)1 and ALDH7 had no effect. These findings implicate TMSNB and PAR2 candidate oncogenes in the pathogenesis of VHL-associated RCC.


Assuntos
Carcinoma de Células Renais/genética , Neoplasias Renais/genética , Proteína Supressora de Tumor Von Hippel-Lindau/genética , Sequência de Bases , Western Blotting , Linhagem Celular , Primers do DNA , Predisposição Genética para Doença , Mutação em Linhagem Germinativa , Humanos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
9.
Plant Biol (Stuttg) ; 20 Suppl 1: 224-230, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28960844

RESUMO

Interactions among plant species via pollinators vary from competitive to mutualistic and can influence the probability of stable coexistence of plant species. We aimed to determine the nature of the interaction via flower visitors between Leucospermum conocarpodendron and Mimetes fimbriifolius, two shrubs in the Proteaceae that share many ecological traits and coexist on the Cape Peninsula, South Africa. To assess the extent of pollinator sharing we analysed nectar properties and recorded the pollinator fauna, their behaviour and contribution to seed set. To test for competition via interspecific pollen transfer, we recorded the movement patterns of pollinators and quantified pollen loads. To determine the effect of co-flowering on visitation rates we recorded visits in stands that varied in the density of the two species. We found that the species produce similar rewards and share pollinating Cape Sugarbirds (Promerops cafer). Interspecific pollen transfer is avoided by placing pollen on different parts of the bird. Both species are visited by nectar-thieving Orange-breasted Sunbirds (Anthobaphes violacea). Insects and autonomous self-pollination contributed little to seed set. Pollinator visits increased with conspecific density in both species, and the slope of the increase was steepest in the presence of high densities of the co-occurring plant species. Nectar thief visits also increased with conspecific density in both species, but the slope declined with increasing density of the co-occurring species. Co-occurrence enhanced pollinator visits and alleviated nectar robbing in both plant species, consistent with mutualisms. Mutualism within a trophic level is unusual, but may help to explain the stable coexistence of ecologically similar species.


Assuntos
Aves/fisiologia , Proteaceae/fisiologia , Simbiose , Animais , Clima , Ecossistema , Insetos/fisiologia , Néctar de Plantas/fisiologia , Polinização/fisiologia , África do Sul
10.
Pediatr Obes ; 13(4): 222-231, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-28296242

RESUMO

BACKGROUND: Restrictive feeding is associated with child overweight; however, the majority of studies used parent-report questionnaires. OBJECTIVES: The relationship between child adiposity measures and directly observed parent and child behaviours were tested using a novel behavioural coding system (BCS). METHODS: Data from 109 children, participants in a twin study and their mothers, were analyzed. Parent-child dyads were video-recorded twice in the laboratory, while children ate ad libitum from a buffet lunch. Mother and child behaviours were assessed using the BCS. Height, body weight and body fat were directly measured for each child. Associations between child adiposity measures and average BCS behaviour (i.e. pooled across visits) were tested using partial correlations adjusting for child age. RESULTS: Regarding discouragement prompts, child body mass index (BMI) z-score was significantly associated with a greater rate of total discouragements (per minute, min-1 ), nonverbal discouragements (min-1 ) and temporary (delay) discouragements (min-1 ) (p < 0.05). Child percent body fat was associated with greater nonverbal discouragements (min-1 ). Regarding encouragement prompts, child BMI z-score was significantly associated with a greater rate of total encouragements (min-1 ), nonverbal encouragements (min-1 ) and reward encouragements (min-1 ). Child BMI z-score and percent body fat were both positively associated with greater maternal health encouragements (min-1 ). Associations with encouragement to eat prompts were no longer significant when accounting for the dependence among twins (being part of the same family). CONCLUSIONS: Heavier children received greater maternal discouragements to eat and, with qualifications, encouragements to eat. The role of nonverbal parenting cues warrants further research regarding child eating regulation and obesity.


Assuntos
Adiposidade , Índice de Massa Corporal , Comportamento Alimentar/psicologia , Relações Pais-Filho , Peso Corporal , Criança , Comportamento Infantil/psicologia , Pré-Escolar , Feminino , Humanos , Masculino , Mães , Obesidade , Sobrepeso , Poder Familiar/psicologia , Obesidade Infantil/psicologia , Inquéritos e Questionários
11.
Diabetes ; 55(3): 667-74, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16505229

RESUMO

The splanchnic bed produces cortisol at rates approximating extraadrenal tissues by converting cortisone to cortisol via the 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 1 pathway. It is not known whether splanchnic cortisol production is regulated by nutrient ingestion and/or by the accompanying changes in hormone secretion. To address this question, 18 healthy humans were randomized to ingest either a mixed meal or to receive an intravenous saline infusion while total-body, splanchnic, and D3 cortisol production (an index of 11beta-HSD type 1 activity) were measured using the combined hepatic catheterization and D4 cortisol infusion methods. Fasting glucose and insulin concentrations did not differ on the meal and saline study days. Glucose and insulin concentrations increased after meal ingestion, peaking at 11.0 +/- 1.0 mmol/l and 451 +/- 64 pmol/l, respectively, at 45 min, then fell to baseline thereafter. In contrast, glucose and insulin concentrations slowly fell to 5.1 +/- 0.1 mmol/l and 27 +/- 6 pmol/l during the 6 h of observation on the saline study day. Fasting cortisol concentration did not differ on the meal and saline study days. Cortisol increased (P < 0.05) to a peak of 353 +/- 55 nmol/l after meal ingestion but did not change after saline infusion. The increase in cortisol after meal ingestion was associated with an increase in both total body cortisol (from 748 +/- 63 to 1,620 +/- 235 nmol/min; P < 0.01) and total body D3 cortisol (from 99 +/- 11 to 143 +/- 11 nmol/min; P < 0.01) production, whereas there was no change in either on the saline study day. The increase in total-body cortisol and D3 cortisol production after meal ingestion originated in extrasplanchnic tissues since splanchnic cortisol production (mean 0-360 min: 254 +/- 83 vs. 262 +/- 36 nmol/min) and splanchnic D3 cortisol production (mean 0-360 min: 72 +/- 22 vs. 77 +/- 14 nmol/min) did not differ on the meal and saline study days. We conclude that ingestion of a mixed meal does not alter either splanchnic cortisol production or the conversion of D4 cortisol to D3 cortisol or, therefore by implication, flux via the splanchnic 11beta-HSD type 1 pathway.


Assuntos
Hidrocortisona/biossíntese , Circulação Esplâncnica , 11-beta-Hidroxiesteroide Desidrogenases/fisiologia , Adulto , Glicemia/análise , Cortisona/biossíntese , Feminino , Artéria Femoral/metabolismo , Alimentos , Veias Hepáticas/metabolismo , Humanos , Insulina/sangue , Masculino , Pessoa de Meia-Idade
12.
J Clin Invest ; 99(8): 1991-8, 1997 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-9109444

RESUMO

To characterize the role of the gap junction protein connexin43 (Cx43) in ventricular conduction, we studied hearts of mice with targeted deletion of the Cx43 gene. Mice homozygous for the Cx43 null mutation (Cx43 -/-) die shortly after birth. Attempts to record electrical activity in neonatal Cx43 -/- hearts (n = 5) were unsuccessful. Ventricular epicardial conduction of paced beats, however, was 30% slower in heterozygous (Cx43 -/+) neonatal hearts (0.14+/-0.04 m/s, n = 27) than in wild-type (Cx43 +/+) hearts (0.20+/-0.07 m/s, n = 32; P < 0.001). This phenotype was even more severe in adult mice; ventricular epicardial conduction was 44% slower in 6-9 mo-old Cx43 -/+ hearts (0.18+/-0.03 m/s, n = 5) than in wild-type hearts (0.32+/-0.07 m/s, n = 7, P < 0.001). Electrocardiograms revealed significant prolongation of the QRS complex in adult Cx43 -/+ mice (13.4+/-1.8 ms, n = 13) compared with Cx43 +/+ mice (11.5+/-1.4 ms, n = 12, P < 0.01). Whole-cell recordings of action potential parameters in cultured disaggregated neonatal ventricular myocytes from Cx43 -/+ and +/+ hearts showed no differences. Thus, reduction in the abundance of a major cardiac gap junction protein through targeted deletion of a Cx43 allele directly leads to slowed ventricular conduction.


Assuntos
Conexina 43/genética , Conexina 43/fisiologia , Sistema de Condução Cardíaco/fisiopatologia , Potenciais de Ação , Animais , Animais Recém-Nascidos , Arritmias Cardíacas/genética , Arritmias Cardíacas/patologia , Arritmias Cardíacas/fisiopatologia , Eletrocardiografia , Feminino , Ventrículos do Coração/patologia , Ventrículos do Coração/fisiopatologia , Heterozigoto , Técnicas In Vitro , Masculino , Camundongos , Camundongos Knockout , Fenótipo
13.
J Clin Invest ; 99(5): 996-1009, 1997 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-9062358

RESUMO

Dystrophic mineralization remains the leading cause of stenotic or regurgitant failure in native human and porcine bioprosthetic heart valves. We hypothesized that cellular expression of noncollagenous matrix proteins (osteopontin, osteocalcin, and osteonectin) that regulate skeletal mineralization may orchestrate valvular calcification. Porcine bioprosthetic heart valves and native human heart valves obtained during replacement surgery were analyzed for cells, matrix proteins that regulate mineralization, and vessels. Cell accumulation and calcification were correlated for both valve types (rho = 0.75, P = 0.01, native; rho = 0.42, P = 0.08, bioprosthetic). Osteopontin expression correlated with cell accumulation (rho = 0.58, P = 0.04) and calcification (rho = 0.52, P = 0.06) for bioprosthetic valves. Osteocalcin expression correlated with calcification (rho = 0.77, P = 0.04) and cell accumulation (rho = 0.69, P = 0.07) in native valves. Comparisons of calcified versus noncalcified native and bioprosthetic valves for averaged total matrix protein mRNA signal score revealed increased noncollagenous proteins mRNA levels in calcified valves (P = 0.07, group I vs. group II; P = 0.02, group III vs. group IV). When stratified according to positive versus negative mRNA signal status, both calcified bioprosthetic valves (P = 0.03) and calcified native valves (P = 0.01) were significantly more positive for noncollagenous proteins mRNA than their noncalcified counterparts. Local cell-associated expression of proteins regulating mineralization suggests a highly coordinated mechanism of bioprosthetic and native valve calcification analogous to physiologic bone mineralization. Modulation of cellular infiltration or cellular expression of matrix proteins that regulate mineralization, may offer an effective therapeutic approach to the prevention of valve failure secondary to calcification.


Assuntos
Bioprótese/efeitos adversos , Calcinose/metabolismo , Cardiomiopatias/metabolismo , Próteses Valvulares Cardíacas/efeitos adversos , Osteocalcina/metabolismo , Osteonectina/metabolismo , Sialoglicoproteínas/metabolismo , Idoso , Animais , Antígenos CD/imunologia , Antígenos de Diferenciação Mielomonocítica/imunologia , Calcificação Fisiológica , Movimento Celular , Sondas de DNA/genética , Biblioteca Gênica , Valvas Cardíacas/química , Valvas Cardíacas/citologia , Valvas Cardíacas/patologia , Humanos , Imuno-Histoquímica , Hibridização In Situ , Macrófagos/imunologia , Osteopontina , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Transdução de Sinais
14.
J Clin Invest ; 76(3): 1229-36, 1985 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-4044833

RESUMO

In order to quantitate the pathways by which liver glycogen is repleted, we administered [1-13C]glucose by gavage into awake 24-h fasted rats and examined the labeling pattern of 13C in hepatic glycogen. Two doses of [1-13C]glucose, 1 and 6 mg/g body wt, were given to examine whether differences in the plasma glucose concentration altered the metabolic pathways via which liver glycogen was replenished. After 1 and 3 h (high-dose group) and after 1 and 2 h (low-dose group), the animals were anesthetized and the liver was quickly freeze-clamped. Liver glycogen was extracted and the purified glycogen hydrolyzed to glucose with amyloglucosidase. The distribution of the 13C-label was subsequently determined by 13C-nuclear magnetic resonance spectroscopy. The percent 13C enrichment of the glucosyl units in glycogen was: 15.1 +/- 0.8%(C-1), 1.5 +/- 0.1%(C-2), 1.2 +/- 0.1%(C-3), 1.1 +/- 0.1%(C-4), 1.6 +/- 0.1%(C-5), and 2.2 +/- 0.1%(C-6) for the high-dose study (n = 4, at 3 h); 16.5 +/- 0.5%(C-1), 2.0 +/- 0.1%(C-2), 1.3 +/- 0.1%(C-3), 1.1 +/- 0.1%(C-4), 2.2 +/- 0.1%(C-5), and 2.4 +/- 0.1%(C-6) in the low-dose study (n = 4, at 2 h). The average 13C-enrichment of C-1 glucose in the portal vein was found to be 43 +/- 1 and 40 +/- 2% in the high- and low-dose groups, respectively. Therefore, the amount of glycogen that was synthesized from the direct pathway (i.e., glucose----glucose-6-phosphate----glucose-1-phosphate----UDP-glucose---- glycogen) was calculated to be 31 and 36% in the high- and low-dose groups, respectively. The 13C-enrichments of portal vein lactate and alanine were 14 and 14%, respectively, in the high-dose group and 11 and 8%, respectively, in the low-dose group. From these enrichments, the minimum contribution of these gluconeogenic precursors to glycogen repletion can be calculated to be 7 and 20% in the high- and low-dose groups, respectively. The maximum contribution of glucose recycling at the triose isomerase step to glycogen synthesis (i.e., glucose----triose-phosphates----glycogen) was estimated to be 3 and 1% in the high- and low-dose groups, respectively. In conclusion, our results demonstrate that (a) only one-third of liver glycogen repletion occurs via the direct conversion of glucose to glycogen, and that (b) only a very small amount of glycogen synthesis can be accounted for by the conversion of glucose to triose phosphates and back to glycogen; this suggests that futile cycling between fructose-6-phosphate and fructose-1,6-diphosphate under these conditions is minimal. Our results also show that (c) alanine and lactate account for a minimum of between 7 and 20% of the glycogen synthesized, and that (d) the three pathways through which the labeled flux is measured account for a total of only 50% of the total glycogen synthesized. These results suggest that either there is a sizeable amount of glycogen synthesis via pathway(s) that were not examined in the present experiment or that there is a much greater dilution of labeled alanine/lactate in the oxaloacetate pool than previously appreciated, or some combination of these two explanations.


Assuntos
Glicogênio Hepático/metabolismo , Fígado/metabolismo , Alanina/sangue , Animais , Glicemia/metabolismo , Cinética , Lactatos/sangue , Fígado/irrigação sanguínea , Glicogênio Hepático/biossíntese , Espectroscopia de Ressonância Magnética , Masculino , Músculos/metabolismo , Veia Porta , Ratos , Ratos Endogâmicos
15.
J Clin Invest ; 78(1): 26-30, 1986 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-3088043

RESUMO

Bone marrow from a normal male pig was transplanted into a related female pig with severe homozygous von Willebrand's disease (vWd). After engraftment the circulating leukocytes were of the male karyotype, and the platelets were strongly positive for von Willebrand factor (vWF) by indirect immunofluorescence. The average level of vWF was 1.96 U/dl and of ristocetin cofactor was 2.8 U/dl. The ear immersion bleeding time before transplantation was consistently more than 15 min and afterwards varied between 5 min and more than 15 min. Transfused vWF corrected the bleeding time at a level of 10 U/dl, which is lower than that required for a von Willebrand pig. We concluded that: the plasmatic compartment is only minimally replenished by the vWF from platelets and megakaryocytes; and the platelet vWF alone only partially corrects the abnormal tests of the hemostatic mechanism in severe vWd.


Assuntos
Transplante de Medula Óssea , Doenças de von Willebrand/terapia , Animais , Antígenos/análise , Tempo de Sangramento , Plaquetas/análise , Transfusão de Sangue , Desamino Arginina Vasopressina/farmacologia , Eletroforese em Gel de Poliacrilamida , Fator VIII/análise , Feminino , Imunofluorescência , Hemostasia , Homozigoto , Cariotipagem , Leucócitos/análise , Masculino , Suínos , Transplante Homólogo , Doenças de von Willebrand/genética , Fator de von Willebrand/análise
16.
J Vet Intern Med ; 31(3): 730-733, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28407319

RESUMO

BACKGROUND: Standard protocols for adrenocorticotropic hormone (ACTH) stimulation testing (ACTHst) often involve intravenous (IV) injection of corticotropin. ACTH might be unintentionally injected into the perivascular (PV) space. OBJECTIVE: To compare stimulation test results after IV and PV injections of ACTH. ANIMALS: Twenty privately owned dogs were studied: 10 healthy and 10 with trilostane-treated naturally occurring hyperadrenocorticism (HAC). METHODS: Prospective study. Each of 20 dogs underwent 2 ACTHst not <4 nor more than 14 days apart. Five healthy and 5 HAC dogs had an IV ACTHst first and PV second; 5 healthy and 5 HAC dogs had a PV ACTHst first and IV second. Blood samples for measurement of serum cortisol concentration were collected before and 1 hour after ACTH administration. RESULTS: No significant difference in results was demonstrated when comparing serum cortisol concentrations after IV and PV ACTH administration in all 20 dogs (median µg/dL; interval µg/dL: 8.2; 1.4-17.4 versus 7.8; 0.9-16.9; P = .23). No significant difference in results was demonstrated when comparing serum cortisol concentrations after IV and PV ACTH administration in the 10 healthy dogs (median µg/dL; interval µg/dL: 10.9; 7.3-17.4 versus 10.6; 7.1-16.9; P = .54) or in the 10 HAC dogs (median µg/dL; interval µg/dL: 6.3; 1.4-8.6 versus 5.2; 0.9-8.7; P = .061). CONCLUSIONS AND CLINICAL IMPORTANCE: Perivascular administration of ACTH does not significantly alter stimulation test results in healthy dogs or in dogs with HAC undergoing therapy with trilostane.


Assuntos
Hiperfunção Adrenocortical/veterinária , Hormônio Adrenocorticotrópico/administração & dosagem , Doenças do Cão/diagnóstico , Hidrocortisona/sangue , Hiperfunção Adrenocortical/diagnóstico , Animais , Estudos de Casos e Controles , Doenças do Cão/sangue , Cães , Feminino , Injeções Intravenosas/veterinária , Injeções Subcutâneas/veterinária , Masculino
17.
Geobiology ; 15(6): 731-749, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28846192

RESUMO

We document the discovery of the first granular iron formation (GIF) of Archaean age and present textural and geochemical results that suggest these formed through microbial iron oxidation. The GIF occurs in the Nconga Formation of the ca. 3.0-2.8 Ga Pongola Supergroup in South Africa and Swaziland. It is interbedded with oxide and silicate facies micritic iron formation (MIF). There is a strong textural control on iron mineralization in the GIF not observed in the associated MIF. The GIF is marked by oncoids with chert cores surrounded by magnetite and calcite rims. These rims show laminated domal textures, similar in appearance to microstromatolites. The GIF is enriched in silica and depleted in Fe relative to the interbedded MIF. Very low Al and trace element contents in the GIF indicate that chemically precipitated chert was reworked above wave base into granules in an environment devoid of siliciclastic input. Microbially mediated iron precipitation resulted in the formation of irregular, domal rims around the chert granules. During storm surges, oncoids were transported and deposited in deeper water environments. Textural features, along with positive δ56 Fe values in magnetite, suggest that iron precipitation occurred through incomplete oxidation of hydrothermal Fe2+ by iron-oxidizing bacteria. The initial Fe3+ -oxyhydroxide precipitates were then post-depositionally transformed to magnetite. Comparison of the Fe isotope compositions of the oncoidal GIF with those reported for the interbedded deeper water iron formation (IF) illustrates that the Fe2+ pathways and sources for these units were distinct. It is suggested that the deeper water IF was deposited from the evolved margin of a buoyant Fe2+aq -rich hydrothermal plume distal to its source. In contrast, oncolitic magnetite rims of chert granules were sourced from ambient Fe2+aq -depleted shallow ocean water beyond the plume.


Assuntos
Sedimentos Geológicos/química , Sedimentos Geológicos/microbiologia , Ferro/química , Essuatíni , Oxirredução , Paleontologia , África do Sul
18.
Clin Obes ; 7(3): 166-175, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28340516

RESUMO

To assess the association of markers for dietary protein intake, measures of dietary adherence and demographic variables with weight loss in the POUNDS Lost study over the first 6 months and again between 6 and 24 months using data from those who completed each period. This is a secondary analysis of pooled data on completers assigned to one of four diets: 65%C/15%P/20%F (AP/LF), 55%C/25%P/20%F (HP/LF), 45%C/15%P/40%F (AP/HF) or 35%C/25%P40%F (HP/HF) in the POUNDS Lost study. Urinary nitrogen excretion, dietary adherence measured by 24-h recall and attendance at sessions, age (above and below 50 years), gender, race/ethnicity and activity by pedometry were analysed. Increased spread between protein intake at baseline and protein at 6 or 24 months, assessed by urinary nitrogen excretion, was associated with greater weight loss from baseline to 2 years. At 6 and 24 months, older age, male gender, body mass index > 30 kg m-2 and adherence to the fat and protein diets were associated with more weight loss. None of these variables was associated with a regain from 6 to 24 months. Weight regain for women in the highest carbohydrate (65%) group was significantly greater (-4.4 kg [95% CI: -5.9, -3.0]) than for women in the lowest carbohydrate group (-1.8 kg [95% CI: -3.2, -0.4 kg]) (P for interaction = 0.012). An increased spread in the difference between baseline and follow-up protein intake was associated with greater weight loss, consistent with the 'protein spread theory'. Women eating the highest carbohydrate diet regained more weight from 6 to 24 months.


Assuntos
Proteínas Alimentares/metabolismo , Obesidade/dietoterapia , Adulto , Idoso , Biomarcadores/metabolismo , Índice de Massa Corporal , Gorduras na Dieta/análise , Gorduras na Dieta/metabolismo , Proteínas Alimentares/análise , Feminino , Humanos , Pessoa de Meia-Idade , Obesidade/metabolismo , Obesidade/fisiopatologia , Obesidade/psicologia , Cooperação do Paciente , Redução de Peso
19.
J Laryngol Otol ; 131(4): 350-356, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28124628

RESUMO

OBJECTIVES: To validate the ovine model of profound oropharyngeal dysphagia and compare swallowing outcomes of laryngotracheal separation with those of total laryngectomy. METHODS: Under real-time fluoroscopy, swallowing trials were conducted using the head and neck of two Dorper cross ewes and one human cadaver, secured in lateral fluoroscopic orientation. Barium trials were administered at baseline, pre- and post-laryngohyoid suspension, following laryngotracheal separation, and following laryngectomy in the ovine model. RESULTS: Mean pre-intervention Penetration Aspiration Scale and National Institutes of Health Swallow Safety Scale scores were 8 ± 0 and 6 ± 0 respectively in sheep and human cadavers, with 100 per cent intra- and inter-species reproducibility. These scores improved to 1 ± 0 and 2 ± 0 post-laryngohyoid suspension (p < 0.01). Aerodigestive tract residue was 18.6 ± 2.4 ml at baseline, 15.4 ± 3.8 ml after laryngotracheal separation and 3.0 ± 0.7 ml after total laryngectomy (p < 0.001). CONCLUSION: The ovine model displayed perfect intra- and inter- species reliability for the Penetration Aspiration Scale and Swallow Safety Scale. Less aerodigestive tract residue after narrow-field laryngectomy suggests that swallowing outcomes after total laryngectomy are superior to those after laryngotracheal separation.


Assuntos
Transtornos de Deglutição/cirurgia , Deglutição/fisiologia , Laringectomia/métodos , Laringoscopia/métodos , Animais , Cadáver , Transtornos de Deglutição/fisiopatologia , Modelos Animais de Doenças , Feminino , Humanos , Laringe/cirurgia , Período Pós-Operatório , Reprodutibilidade dos Testes , Índice de Gravidade de Doença , Ovinos , Traqueia/cirurgia , Resultado do Tratamento
20.
Geobiology ; 14(3): 255-75, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26750514

RESUMO

Chocolate Pots hot springs (CP) is a unique, circumneutral pH, iron-rich, geothermal feature in Yellowstone National Park. Prior research at CP has focused on photosynthetically driven Fe(II) oxidation as a model for mineralization of microbial mats and deposition of Archean banded iron formations. However, geochemical and stable Fe isotopic data have suggested that dissimilatory microbial iron reduction (DIR) may be active within CP deposits. In this study, the potential for microbial reduction of native CP Fe(III) oxides was investigated, using a combination of cultivation dependent and independent approaches, to assess the potential involvement of DIR in Fe redox cycling and associated stable Fe isotope fractionation in the CP hot springs. Endogenous microbial communities were able to reduce native CP Fe(III) oxides, as documented by most probable number enumerations and enrichment culture studies. Enrichment cultures demonstrated sustained DIR driven by oxidation of acetate, lactate, and H2 . Inhibitor studies and molecular analyses indicate that sulfate reduction did not contribute to observed rates of DIR in the enrichment cultures through abiotic reaction pathways. Enrichment cultures produced isotopically light Fe(II) during DIR relative to the bulk solid-phase Fe(III) oxides. Pyrosequencing of 16S rRNA genes from enrichment cultures showed dominant sequences closely affiliated with Geobacter metallireducens, a mesophilic Fe(III) oxide reducer. Shotgun metagenomic analysis of enrichment cultures confirmed the presence of a dominant G. metallireducens-like population and other less dominant populations from the phylum Ignavibacteriae, which appear to be capable of DIR. Gene (protein) searches revealed the presence of heat-shock proteins that may be involved in increased thermotolerance in the organisms present in the enrichments as well as porin-cytochrome complexes previously shown to be involved in extracellular electron transport. This analysis offers the first detailed insight into how DIR may impact the Fe geochemistry and isotope composition of a Fe-rich, circumneutral pH geothermal environment.


Assuntos
Bactérias/metabolismo , Compostos Férricos/metabolismo , Fontes Termais/microbiologia , Bactérias/classificação , Oxirredução , Parques Recreativos , RNA Bacteriano/análise , RNA Ribossômico 16S/análise , Wyoming
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