Vernalization, the acceleration of flowering by a long period of cold temperature, ensures that many plants overwinter vegetatively and flower in spring. In Arabidopsis, allelic variation at the ...FRIGIDA (FRI) locus is a major determinant of natural variation in flowering time. Dominant alleles of FRI confer late flowering, which is reversed to earliness by vernalization. We cloned FRI and analyzed the molecular basis of the allelic variation. Most of the early-flowering ecotypes analyzed carry FRI alleles containing one of two different deletions that disrupt the open reading frame. Loss-of-function mutations at FRI have thus provided the basis for the evolution of many early-flowering ecotypes.
A study was made of the antioxidant content, activity and colour of two New Zealand commercially grown tomatoes (
Lycopersicon esculentum L
. var. Excell and Aranca) cultivated in glasshouses using ...hydroponic techniques. Excell tomatoes were harvested and stored individually while cultivar Aranca was harvested by cutting the vine, which consisted of a group of eight tomatoes. Both cultivars of tomatoes were stored in the dark for 4 days at 15°C to simulate normal pre-purchase storing conditions. The antioxidant content of the raw tomatoes after 4 days of storage were markedly different while the CIE LAB colour values of the cut inner surfaces of the two cultivars were similar. After 4 days storage, subsamples of each cultivar were either boiled, baked or fried, and analysis of CIE colour, ascorbic acid, total phenolics, lycopene and antioxidant activity (using the ABTS assay) was undertaken. Boiling and baking had a relatively small effect on the ascorbic, total phenolic, lycopene and antioxidant activity of the two cultivars while frying significantly reduced (
P<0.001) the ascorbic, total phenolic and lycopene contents of the two cultivars. Chromatic colour analysis showed that both cultivars became significantly (
P<0.001) darker and less red after cooking by all methods.
In a following experiment, the two cultivars of tomatoes were sliced and allowed to soak for 20
min in a mixture of olive oil and white vinegar, or olive oil and white vinegar separately. CIE colour of the two cultivars showed no change after processing but the treatments of oil and vinegar separately and together reduced (
P<0.05) the red component of the colour. Treating the two cultivars of tomatoes with the oil and vinegar mixture resulted in a significant reduction (
P<0.001) in the ascorbic acid, total phenolic and antioxidant activity of the tomatoes. Treatment with oil significantly reduced the amount of lycopene that could be extracted from the tomatoes while treatment with vinegar had no effect.
In an atom, the static external electric field generated by the tiny central nuclear charge is spherical, so the cloud of electrons that defines its chemical and mechanical properties is always ...spherical. With new accelerators being built around the world with the aim of producing beams of exotic isotopes, and ever more sensitive detectors for measuring electromagnetic radiation patterns, we can expect more of this 'isotope tailoring' followed by Coulomb excitation.
Centromere protein CENP-E is a dimeric kinesin (Kinesin-7 family) with critical roles in mitosis, including establishment of microtubule (MT)-chromosome linkage and movement of mono-oriented ...chromosomes on kinetochore microtubules for proper alignment at metaphase 1–9. We performed studies to test the hypothesis that CENP-E promotes MT elongation at the MT plus ends. A human CENP-E construct was engineered, expressed, and purified, and it yielded the CENP-E-6His dimeric motor protein. The results show that CENP-E promotes MT plus-end-directed MT gliding at 11 nm/s. The results from real-time microscopy assays indicate that 60.3% of polarity-marked MTs exhibited CENP-E-promoted MT plus-end elongation. The MT extension required ATP turnover, and MT plus-end elongation occurred at 1.48 μm/30 min. Immunolocalization studies revealed that 80.8% of plus-end-elongated MTs showed CENP-E at the MT plus end. The time dependence of CENP-E-promoted MT elongation in solution best fit a single exponential function (kobs = 5.1 s−1), which is indicative of a mechanism in which α,β-tubulin subunit addition is tightly coupled to ATP turnover. Based on these results, we propose that CENP-E, as part of its function in chromosome kinetochore-MT linkage, plays a direct role in MT elongation.
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► CENP-E promotes MT plus-end elongation ► CENP-E localizes to the plus end of elongated MTs ► CENP-E-promoted MT elongation is tightly coupled to ATP turnover ► Model: CENP-E steps to the MT plus end, stabilizing the MT straight-end conformation
The fluorescent non-canonical amino acid tagging (FUNCAT) technique has been used to visualize newly synthesized proteins in cell lines and tissues. Here, we present a protocol for measuring protein ...synthesis in specific cell types in the mouse brain using in vivo FUNCAT. We describe steps for metabolically labeling newly synthesized proteins with azidohomoalanine, which introduces an azide group into the polypeptide. We then detail procedures for binding a fluorophore-conjugated alkyne to the azide group to allow its visualization.
For complete details on the use and execution of this protocol, please refer to tom Dieck et al. (2012)1 and Hooshmandi et al. (2023).2
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•Steps for L-azidohomoalanine (AHA) incorporation into the nascent proteins in vivo•Cell-type-specific visualization of de novo protein synthesis in the brain•AHA signal intensity quantification and analysis in distinct neuronal cell types
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
The fluorescent non-canonical amino acid tagging (FUNCAT) technique has been used to visualize newly synthesized proteins in cell lines and tissues. Here, we present a protocol for measuring protein synthesis in specific cell types in the mouse brain using in vivo FUNCAT. We describe steps for metabolically labeling newly synthesized proteins with azidohomoalanine, which introduces an azide group into the polypeptide. We then detail procedures for binding a fluorophore-conjugated alkyne to the azide group to allow its visualization.
Background:
There are approximately 2 million people living with the loss of a major limb in America. It is estimated that 95% of these will have some form of pain associated with their amputation. ...Phantom limb pain, related to symptomatic neuromas, contributes to amputation morbidity and can be difficult to treat. Studies have shown that targeted muscle reinnervation (TMR), by giving symptomatic neuromas “somewhere to go and something to do,” can be an effective therapy. However, a large proportion of surgeons still treat symptomatic neuromas by burying them in nearby tissue.
Methods:
We treated a patient with previous above-the-knee amputation, complicated by a symptomatic neuroma, with TMR. We identified and described nine steps to the procedure. Our description is accompanied by illustrative, intraoperative photographs and technical pearls.
Results:
This article provides a description of TMR technique involving a neuroma of the sciatic nerve and its branches, to treat an above-the-knee amputation, with the aim of making this approach more accessible. At 9-month follow-up, the patient had active firing of the recipient muscles with donor nerve stimulation indicating successful reinnervation. The patient continued to report stump pain, but with intermittent pain-free days.
Conclusions:
TMR has proven potential as a therapy for amputation-related, neuropathic pain. With this technical guide to TMR, surgeons should feel more comfortable adding this technique to their armamentarium, to be utilized either at the time of amputation or as a secondary measure.
Abstract
Abstract
How can people achieve successful communication when using novel signs? Previous studies show that iconic signs (i.e. signs that directly resemble their referent) enhance ...communication success. In this paper, we test if enculturated signs (i.e. signs informed by interlocutors’ shared culture) also enhance communication success. Children, who have spent less time in their linguistic community, have less cultural knowledge to inform their sign innovation. A natural prediction is that younger children's signs will be less enculturated, more diverse and less successful compared with older children and adults. We examined sign innovation in children aged between 6 and 12 years (
N
= 54) and adults (
N
= 18). Sign enculturation, diversity and iconicity were rated. As predicted, younger children innovated less enculturated and more diverse signs, and communicated less successfully than older children and adults. Sign enculturation and iconicity uniquely contributed to communication success. This is the first study to demonstrate that enculturated signs enhance communication.
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