92 research outputs found
Preparation of Large Monodisperse Vesicles
Preparation of monodisperse vesicles is important both for research purposes and for practical applications. While the extrusion of vesicles through small pores (∼100 nm in diameter) results in relatively uniform populations of vesicles, extrusion to larger sizes results in very heterogeneous populations of vesicles. Here we report a simple method for preparing large monodisperse multilamellar vesicles through a combination of extrusion and large-pore dialysis. For example, extrusion of polydisperse vesicles through 5-µm-diameter pores eliminates vesicles larger than 5 µm in diameter. Dialysis of extruded vesicles against 3-µm-pore-size polycarbonate membranes eliminates vesicles smaller than 3 µm in diameter, leaving behind a population of monodisperse vesicles with a mean diameter of ∼4 µm. The simplicity of this method makes it an effective tool for laboratory vesicle preparation with potential applications in preparing large monodisperse liposomes for drug delivery
A Triple Protostar System Formed via Fragmentation of a Gravitationally Unstable Disk
Binary and multiple star systems are a frequent outcome of the star formation
process, and as a result, almost half of all sun-like stars have at least one
companion star. Theoretical studies indicate that there are two main pathways
that can operate concurrently to form binary/multiple star systems: large scale
fragmentation of turbulent gas cores and filaments or smaller scale
fragmentation of a massive protostellar disk due to gravitational instability.
Observational evidence for turbulent fragmentation on scales of 1000~AU has
recently emerged. Previous evidence for disk fragmentation was limited to
inferences based on the separations of more-evolved pre-main sequence and
protostellar multiple systems. The triple protostar system L1448 IRS3B is an
ideal candidate to search for evidence of disk fragmentation. L1448 IRS3B is in
an early phase of the star formation process, likely less than 150,000 years in
age, and all protostars in the system are separated by 200~AU. Here we
report observations of dust and molecular gas emission that reveal a disk with
spiral structure surrounding the three protostars. Two protostars near the
center of the disk are separated by 61 AU, and a tertiary protostar is
coincident with a spiral arm in the outer disk at a 183 AU separation. The
inferred mass of the central pair of protostellar objects is 1 M,
while the disk surrounding the three protostars has a total mass of 0.30
M_{\sun}. The tertiary protostar itself has a minimum mass of 0.085
M. We demonstrate that the disk around L1448 IRS3B appears susceptible
to disk fragmentation at radii between 150~AU and 320~AU, overlapping with the
location of the tertiary protostar. This is consistent with models for a
protostellar disk that has recently undergone gravitational instability,
spawning one or two companion stars.Comment: Published in Nature on Oct. 27th. 24 pages, 8 figure
Farmers’ Perception of Viral Diseases and Their Management in Pepper (Capsicum spp.) Production in Benin
Pepper (Capsicum spp.) is an important solanaceous cash crop in Benin; however, productivity is limited due to several key constraints, especially diseases caused by viruses. We sought to understand farmers’ perceptions of viral diseases, management strategies deployed, and to identify the virus population affecting pepper production in Benin. To assess farmers’ perceptions and management of viral diseases, a survey was carried out in four agroecological zones of Benin. A total of 144 pepper farmers were interviewed using the snowball method. A total of 52 pepper leaf samples with virus-like symptoms were collected and diagnosed by reverse-transcriptase polymerase chain reaction (RT-PCR) or PCR. Pepper production systems varied across agroecological zones (P ≤ 0.001) with a predominance of farms practicing monoculture (82%). The majority of farmers (89%) indicated that pests and diseases were the main constraints to increased production. Cucumber mosaic virus (92% of the total samples), Pepper vein yellow virus (52%), and Pepper veinal mottle virus (50%) were the major viruses detected in pepper fields in Benin. There were both single (29%) and mixed (71%) infections of the viruses, suggesting that mixed infections are common for pepper in Benin, confounding efforts to reduce virus infections. Nearly 100% of the farmers surveyed were not aware of these viral diseases. They also could not directly relate symptoms of virus infection to the presence of aphids, whiteflies, or thrips. Farmers relied primarily on synthetic insecticides (93%) to control virus vectors. Interestingly, some farmers applied commercial (12%) and homemade (17%) biopesticides, with neem-based preparations being the most widely used. A total of 15% of farmers used companion cropping with maize, mint or basil and 43% of farmers used crop rotation as a cultural management practice to control viral disease and vector pressure in pepper fields. The implications of this work include the importance of training farmers and extension agents on diagnosis of viruses and their vectors causing viral diseases. This study provides baseline information for the development of host-resistant cultivars and deployment of integrated pest management strategies for pepper in Benin to reduce farmer losses
Redundant Mechanisms Prevent Mitotic Entry Following Replication Arrest in the Absence of Cdc25 Hyper-Phosphorylation in Fission Yeast
Following replication arrest the Cdc25 phosphatase is phosphorylated and inhibited by Cds1. It has previously been reported that expressing Cdc25 where 9 putative amino-terminal Cds1 phosphorylation sites have been substituted to alanine results in bypass of the DNA replication checkpoint. However, these results were acquired by expression of the phosphorylation mutant using a multicopy expression vector in a genetic background where the DNA replication checkpoint is intact. In order to clarify these results we constructed a Cdc25(9A)-GFP native promoter integrant and examined its effect on the replication checkpoint at endogenous expression levels. In this strain the replication checkpoint operates normally, conditional on the presence of the Mik1 kinase. In response to replication arrest the Cdc25(9A)-GFP protein is degraded, suggesting the presence of a backup mechanism to eliminate the phosphatase when it cannot be inhibited through phosphorylation
Biochemical Comparison of Anopheles gambiae and Human NADPH P450 Reductases Reveals Different 2′-5′-ADP and FMN Binding Traits
NADPH-cytochrome P450 oxidoreductase (CPR) plays a central role in chemical
detoxification and insecticide resistance in Anopheles gambiae,
the major vector for malaria. Anopheles gambiae CPR (AgCPR) was
initially expressed in Eschericia coli but failed to bind
2′, 5′-ADP Sepharose. To investigate this unusual trait, we
expressed and purified a truncated histidine-tagged version for side-by-side
comparisons with human CPR. Close functional similarities were found with
respect to the steady state kinetics of cytochrome c reduction,
with rates (kcat) of 105
s−1 and 88 s−1, respectively, for mosquito
and human CPR. However, the inhibitory effects of 2′,5′-ADP on
activity were different; the IC50 value of AgCPR for 2′,
5′ –ADP was significantly higher (6–10 fold) than human CPR
(hCPR) in both phosphate and phosphate-free buffer, indicative of a decrease in
affinity for 2′, 5′- ADP. This was confirmed by isothermal titration
calorimetry where binding of 2′,5′-ADP to AgCPR
(Kd = 410±18 nM) was
∼10 fold weaker than human CPR
(Kd = 38 nM). Characterisation
of the individual AgFMN binding domain revealed much weaker binding of FMN
(Kd = 83±2.0 nM) than the equivalent
human domain (Kd = 23±0.9 nM).
Furthermore, AgCPR was an order of magnitude more sensitive than hCPR to the
reductase inhibitor diphenyliodonium chloride
(IC50 = 28 µM±2 and 361±31
µM respectively). Taken together, these results reveal unusual biochemical
differences between mosquito CPR and the human form in the binding of small
molecules that may aid the development of ‘smart’ insecticides and
synergists that selectively target mosquito CPR
Inhibition of StearoylCoA Desaturase-1 Inactivates Acetyl-CoA Carboxylase and Impairs Proliferation in Cancer Cells: Role of AMPK
Cancer cells activate the biosynthesis of saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) in order to sustain an increasing demand for phospholipids with appropriate acyl composition during cell replication. We have previously shown that a stable knockdown of stearoyl-CoA desaturase 1 (SCD1), the main Δ9-desaturase that converts SFA into MUFA, in cancer cells decreases the rate of lipogenesis, reduces proliferation and in vitro invasiveness, and dramatically impairs tumor formation and growth. Here we report that pharmacological inhibition of SCD1 with a novel small molecule in cancer cells promoted the activation of AMP-activated kinase (AMPK) and the subsequent reduction of acetylCoA carboxylase activity, with a concomitant inhibition of glucose-mediated lipogenesis. The pharmacological inhibition of AMPK further decreased proliferation of SCD1-depleted cells, whereas AMPK activation restored proliferation to control levels. Addition of supraphysiological concentrations of glucose or pyruvate, the end product of glycolysis, did not reverse the low proliferation rate of SCD1-ablated cancer cells. Our data suggest that cancer cells require active SCD1 to control the rate of glucose-mediated lipogenesis, and that when SCD1 activity is impaired cells downregulate SFA synthesis via AMPK-mediated inactivation of acetyl-CoA carboxylase, thus preventing the harmful effects of SFA accumulation
Perceived social support disparities among children affected by HIV/AIDS in Ghana: a cross-sectional survey
Variability of Brown Dwarfs
Brown dwarfs constitute a missing link between low-mass stars and giant
planets. Their atmospheres display chemical species typical of planets, and one
could wonder whether they also have weather-like patterns. While brown dwarf
surface features cannot be directly resolved, the photometric and spectroscopic
modulations induced by these features, as they rotate in and out of view,
provide a wealth of information on the evolution of their atmosphere. A review
of brown dwarfs variability through the L, T and Y spectral types sequence is
presented, as well as the constraints that they set on the nature of
weather-like patterns on their surface.Comment: Accepted chapter in the "Handbook of Exoplanets"; Springe
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Full field electroretinogram in autism spectrum disorder
Purpose
To explore early findings that individuals with autism spectrum disorder (ASD) have reduced scotopic ERG b-wave amplitudes.
Methods
Dark adapted (DA) ERGs were acquired to a range of flash strengths, (-4.0 to 2.3 log phot cd.s.m-2), including and extending the ISCEV standard, from two subject groups: (ASD) N=11 and (Control) N=15 for DA and N=14 for light adapted (LA) ERGs who were matched for mean age and range. Naka-Rushton curves were fitted to DA b-wave amplitude growth over the first limb (-4.0 to -1.0 log phot cd.s.m-2). The derived parameters (Vmax, Km and n) were compared between groups. Scotopic 15 Hz flicker ERGs (14.93Hz) were recorded to 10 flash strengths presented in ascending order from -3.0 to 0.5 log Td.s to assess the slow and fast rod pathways respectively. LA ERGs were acquired to a range of flash strengths, (-0.5 to 1.0 log phot cd.s.m-2). Photopic 30 Hz, flicker ERGs, oscillatory potentials (OPs) and the responses to prolonged 120 ms ON- OFF stimuli were also recorded.
Results
For some individuals the DA b-wave amplitudes fell below the control 5th centile of the controls with up to four ASD participants (36%) at the 1.5 log phot cd.s.m-2 flash strength and two (18%) ASD participants at the lower -2 log phot cd.s.m-2 flash strength. However, across the thirteen flash strengths there were no significant group differences for b-wave amplitude’s growth (repeated measures ANOVA p=0.83). Nor were there any significant differences between the groups for the Naka-Rushton parameters (p>0.09). No group differences were observed in the 15Hz scotopic flicker phase or amplitude (p>0.1), DA ERG a- wave amplitude or time to peak (p>26). The DA b-wave time to peak at 0.5 log phot cd.s.m-2 were longer in the ASD group (corrected p=0.04). The single ISCEV LA 0.5 log phot cd.s.m-2 (p0.08) to the single flash stimuli although there was a significant interaction between group and flash strength for the b-wave amplitude (corrected p=0.006). The prolonged 120 ms ON-responses were smaller in the ASD group (corrected p=0.003), but the OFF response amplitude (p>0.6) and ON and OFF times to peaks (p>0.4) were similar between groups. The LA OPs showed an earlier bifurcation of OP2 in the younger ASD participants, however no other differences were apparent in the OPs or 30Hz flicker waveforms.
Conclusion
Some ASD individuals show subnormal DA ERG b-wave amplitudes. Under LA conditions the b-wave is reduced across the ASD group along with the ON response of the ERG. These exploratory findings, suggest there is altered cone-ON bipolar signalling in ASD
The LeucoPatch® system in the management of hard-to-heal diabetic foot ulcers: study protocol for a randomised controlled trial
Background: Diabetic foot ulcers are a common and severe complication of diabetes mellitus. Standard treatment includes debridement, offloading, management of infection and revascularisation where appropriate, although healing times may be long. The LeucoPatch® device is used to generate an autologous platelet-rich fibrin and leucocyte wound dressing produced from the patient's own venous blood by centrifugation, but without the addition of any reagents. The final product comprises a thin, circular patch composed predominantly of fibrin together with living platelets and leucocytes. Promising results have been obtained in non-controlled studies this system, but this now needs to be tested in a randomised controlled trial (RCT). If confirmed, the LeucoPatch® may become an important new tool in the armamentarium in the management of diabetic foot ulcers which are hard-to-heal. Methods: People with diabetes and hard-to-heal ulcers of the foot will receive either pre-specified good standard care or good standard care supplemented by the application of the LeucoPatch® device. The primary outcome will be the percentage of ulcers healed within 20 weeks. Healing will be defined as complete epithelialisation without discharge that is maintained for 4 weeks and is confirmed by an observer blind to randomisation group. Discussion: Ulcers of the foot are a major source of morbidity to patients with diabetes and costs to health care economies. The study population is designed to be as inclusive as possible with the aim of maximising the external validity of any findings. The primary outcome measure is healing within 20 weeks of randomisation and the trial also includes a number of secondary outcome measures. Among these are rate of change in ulcer area as a predictor of the likelihood of eventual healing, minor and major amputation of the target limb, the incidence of infection and quality of life. Trial registration: International Standard Randomised Controlled Trial, ISRCTN27665670. Registered on 5 July 2013
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