Red Form Research Articles

Modulation of green to red photoconversion of GFP during fluorescent microscopy by carbon source and oxygen availability.

Studies have reported on the ability of green fluorescent proteins to photoconvert into a red fluorescent form under various conditions, such as the presence of oxidants, hypoxia, as well as under benign conditions using irradiation with a 405 nm laser. Here, we show that in Saccharomyces cerevisiae yeast green fluorescent protein (GFP) (S65T) fused to different cellular proteins can easily photoconvert into a red form when cells are grown in media with nonfermentable carbon sources. This photoconversion occurs during standard microscopy between glass slide and coverslip but is completely prevented by imaging on pads of solid medium or in a large volume of medium on an inverted microscope. The observed effect was due to rapid hypoxia of cells with respiratory metabolism in standard conditions for upright microscopy. Photoconversion could be prevented by antioxidant treatment, suggesting that it proceeds via the effects of reactive oxidative species emerging in response to oxygen deficiency. Our results show the need for caution during upright microscopy imaging in conditions where there is active respiration and demonstrate simple approaches to prevent unwanted GFP photoconversion. They also provide easy means of performing photoconversion experiments on existing GFP-bearing cell lines, at least in the case of yeast.

Patient Initiated Side Effects Reporting System in their Vernacular Language: A Cross Sectional Analysis of Feasibility and Acceptability in Our Scenario

<p style="text-align: justify;"><strong>Objectives:</strong> To determine the acceptability of Patient initiated side effects reporting system in their vernacular language. <strong>Methods:</strong> All the hundred participants were using simple non-probability sampling; participants were given red form and blue form 9 Central drug standard and control suspected adverse effects form for physicians and patients respectively), placed them comfortably and asked to compare and their answers were coded when there were ready to convey the answers. All the questions were closed ended with prior testing with Cronbach alpha value of 0.75 to find suitability, both sheets were give and time allotted for them to get acquaintainted and then questions were put forth. <strong>Results: </strong>Nearly 65 subjects were aware that even minor side effects should be reported in blue form by themselves while 35% percentage was not familiar with it. Another significant finding about 66% people was not familiar with their names or identity such as revealing it in the form. <strong>Conclusion:</strong> Having adverse drug reaction from and measures taken were laudable; however, suitability and feasibility in our Indian scenario should be improved. Population education may play a key role and appointing social pharmacist is also a key option to alleviate ground level disconnects. <p style="text-align: justify;"><strong>Key words: </strong>Patient initiated adverse drug reporting, Consumer side effects reporting from, Blue and red from.

Population genetic structure underlying the geographic variation in beetle structural colour with multiple transition zones.

We studied the population genetic structure underlying the geographic variation in the structural colour of the geotrupid dung beetle, Phelotrupes auratus, which exhibits metallic body colours of different reflectance wavelengths perceived as red, green and indigo. These forms occur parapatrically in an area of Japan. The colour variation was not related to variation in climatic factors. Using single nucleotide polymorphisms (SNPs) from restriction-site-associated DNA sequences, we discriminated five groups of populations (west/red, south/green, south/indigo, south/red and east/red) by a combination of genetic clusters (west, south and east) and three colour forms. There were three transition zones for the colour forms: two between the red and green forms were hybrid zones with steep genetic clines, which implies the existence of barriers to gene flow between regions with different colours. The remaining transition zone between the green and indigo forms lacked genetic differentiation, despite the evident colour changes, which implies regionally specific selection on the different colours. In a genomewide association study, we identified four SNPs that were associated with the red/green or indigo colour and were not linked with one another, which implies that the coloration was controlled by multiple loci, each affecting the expression of a different colour range. These loci may have controlled the transitions between different combinations of colours. Our study demonstrates that geographic colour variation within a species can be maintained by nonuniform interactions among barriers to gene flow, locally specific selection on different colours, and the effects of different colour loci.

IDENTIFICATION PLASTIC BOTTLE BASED ON LIGHTING SYSTEM

The challenge in waste sorting systems such as plastic bottles is how to identify and classify the types of it. This work aims to build a system of identification and classification of sorting plastic bottles by type. This system uses the Backpropagation method by utilizing a webcam to get colors in the form of Red, Green, and Blue (RGB) color spaces. As light control, three types of lights are used, blue, red, and yellow. The three types of plastics that will be identified are PET, HDPE, and PP. The conclusion from this work is that the percentage of success is 18%, 28%, and 42 for yellow, blue, and red lighting, respectively.

Using spent tea residue to elimination of chromium in aqueous liquid

In this work, spent red and black tea form leaves were assessed for their ability to doff expel chromium ion from aqueous solutions at various values of PH. Classification adsorption experiments indebted at choice temperature and showed cruise the adsorption of Cr ion decrease more the increase of temperature. In successive lyrics, the adsorption fight is exothermic and the enthalpy is fatal, at long last the unconventional ways and the entropy are positive. The matter copied at equal stat was compared with Langmuir, Freundlich models, and the inference efficiencies for Cr ion higher than 99% and can easily be achieved. It was hinge saunter the brilliant hatch is reform than hyacinthine cause in adsorption of chromium ion.

Environmentally Sensitive Color‐Shifting Fluorophores for Bioimaging

AbstractWe introduce color‐shifting fluorophores that reversibly switch between a green and red fluorescent form through intramolecular spirocyclization. The equilibrium of the spirocyclization is environmentally sensitive and can be directly measured by determining the ratio of red to green fluorescence, thereby enabling the generation of ratiometric fluorescent probes and biosensors. Specifically, we developed a ratiometric biosensor for imaging calcium ions (Ca2+) in living cells, ratiometric probes for different proteins, and a bioassay for the quantification of nicotinamide adenine dinucleotide phosphate.

Environmentally Sensitive Color-Shifting Fluorophores for Bioimaging.

We introduce color‐shifting fluorophores that reversibly switch between a green and red fluorescent form through intramolecular spirocyclization. The equilibrium of the spirocyclization is environmentally sensitive and can be directly measured by determining the ratio of red to green fluorescence, thereby enabling the generation of ratiometric fluorescent probes and biosensors. Specifically, we developed a ratiometric biosensor for imaging calcium ions (Ca2+) in living cells, ratiometric probes for different proteins, and a bioassay for the quantification of nicotinamide adenine dinucleotide phosphate.

On the spectral properties and excitation dynamics of long-wavelength chlorophylls in higher-plant photosystem I

In higher-plant Photosystem I (PSI), the majority of “red” chlorophylls (absorbing at longer wavelengths than the reaction centre P700) are located in the peripheral antenna, but contradicting reports are given about red forms in the core complex. Here we attempt to clarify the spectroscopic characteristics and quantify the red forms in the PSI core complex, which have profound implication on understanding the energy transfer and charge separation dynamics. To this end we compare the steady-state absorption and fluorescence spectra and picosecond time-resolved fluorescence kinetics of isolated PSI core complex and PSI–LHCI supercomplex from Pisum sativum recorded at 77 K. Gaussian decomposition of the absorption spectra revealed a broad band at 705 nm in the core complex with an oscillator strength of three chlorophylls. Additional absorption at 703 nm and 711 nm in PSI–LHCI indicated up to five red chlorophylls in the peripheral antenna. Analysis of fluorescence emission spectra resolved states emitting at 705, 715 and 722 nm in the core and additional states around 705–710 nm and 733 nm in PSI–LHCI. The red states compete with P700 in trapping excitations in the bulk antenna, which occurs on a timescale of ~20 ps. The three red forms in the core have distinct decay kinetics, probably in part determined by the rate of quenching by the oxidized P700. These results affirm that the red chlorophylls in the core complex must not be neglected when interpreting kinetic experimental results of PSI.

Comparative anti-inflammatory evaluation of five plants of Solanaceae members

Inflammation is processed by which the body responds to the external stimulus that causes any harm or damage to the human body. Inflammation is good with some respect and if it grows and builds up in the body will lead to many other complications and manifests itself in the form of pain, redness, swelling and spike in surrounding tissue temperature. Solanaceae members are known to have rich chemical constituents and a variety of them being alkaloids. Whatever may be the chemical constituent, most of the Solanaceae members have been already investigated and proven for their anti-inflammatory activity in many available methods. In research, all five species of Solanum had been compared for their anti-inflammatory activity in two models to determine which has a better activity in comparison to the other. Overall it can be said that Solanum nigrum showed a better activity compared to other extracts and the activity was comparably significant and similar to that of the standard drugs dexamethasone and indomethacin too.

Push–Pull (Iso)quinoline Chromophores: Synthesis, Photophysical Properties, and Use for White‐Light Emission

The photophysical properties of a series of conjugated push-pull (iso)quinolines were studied. The compounds were synthesized by well-established and straightforward methodologies. The materials exhibited not only emission solvatochromism in a variety of nonpolar solvents, but also tunable halochromism. Some of the compounds remained moderately luminescent after protonation and had a red emissive form, which was used to obtain white-light emission, both in solution and in thin films, by controlled protonation of the initially blue-green-emitting materials. This methodology has potential applications in the fabrication of white organic light-emitting diodes with two forms of a single emitter in equilibrium.

Freeze-dried celery as an indirect source of nitrate in cold-smoked sausages: Effect on safety and color formation

The study investigated the possibility of changing synthetic nitrate and nitrite to nitrate from freeze-dried celery in cold-smoked sausages in order to improve their quality and safety. Cold-smoked sausages containing 2.58% of freeze-dried celery (corresponding to 150 mg/kg of nitrate) or 150 mg/kg nitrite or 150 mg/kg nitrate were manufactured with S. xylosus or with S. xylosus and P. pentosaceus mixture. Such characteristics as physicochemical properties, microbiological quality, and the relative changes of myoglobin forms were evaluated during the fermentation and ripening processes. Synthetic nitrite or nitrate replacement with nitrate from freeze-dried celery in cold-smoked sausages influenced the lower pH values (5.04–5.10). The residual nitrate content in sausage depended on the type of nitrate and starter cultures used in the sausages production. The total quantity of red myoglobin forms (oxymyoglobin and deoxymyoglobin) was very similar in all cold-smoked sausages; however, the lower redness values (8.50–9.32) were observed in sausages with freeze-dried celery.The results of the presented study indicate that freeze-dried celery could be used instead of added nitrite while maintaining the same quality and safety of cold-smoked sausages. However, it is essential to select the starter cultures properly in order to ensure controlled fermentation.

Working the red carpet: a framework for analysing celebrities’ red carpet labour

ABSTRACT This article critically assesses celebrities’ labour on the red carpet, with particular attention to the ways in which they engage with a variety of social and political issues. Through a discursive analysis of three 2019 red carpet events, we explore how the norms and values of the red carpet demand different forms of aesthetic and emotional labour from celebrities. We specifically theorise the concept red carpet labour as the multiple intersecting forms of work that celebrities negotiate on the red carpet, which create and constrain the possibilities for them to intervene in broader cultural conversations during their red carpet appearances. These intersecting forms of labour are laid out in a three-part conceptual framework that argues for scholarly attention to how celebrities (1) participate in the construction of red carpets as pseudoevents, (2) work to create a self-brand on the red carpet, and (3) cultivate relationships with the fashion world.

Harvesting far-red light: Functional integration of chlorophyll f into Photosystem I complexes of Synechococcus sp. PCC 7002

The heterologous expression of the far-red absorbing chlorophyll (Chl) f in organisms that do not synthesize this pigment has been suggested as a viable solution to expand the solar spectrum that drives oxygenic photosynthesis. In this study, we investigate the functional binding of Chl f to the Photosystem I (PSI) of the cyanobacterium Synechococcus 7002, which has been engineered to express the Chl f synthase gene. By optimizing growth light conditions, one-to-four Chl f pigments were found in the complexes. By using a range of spectroscopic techniques, isolated PSI trimeric complexes were investigated to determine how the insertion of Chl f affects excitation energy transfer and trapping efficiency. The results show that the Chls f are functionally connected to the reaction center of the PSI complex and their presence does not change the overall pigment organization of the complex. Chl f substitutes Chl a (but not the Chl a red forms) while maintaining efficient energy transfer within the PSI complex. At the same time, the introduction of Chl f extends the photosynthetically active radiation of the new hybrid PSI complexes up to 750 nm, which is advantageous in far-red light enriched environments. These conclusions provide insights to engineer the photosynthetic machinery of crops to include Chl f and therefore increase the light-harvesting capability of photosynthesis.

Effect of circularly polarized light on germination, hypocotyl elongation and biomass production of arabidopsis and lettuce: Involvement of phytochrome B.

Circular dichroism (CD), defined as the differential absorption of left- and right-handed circularly polarized light (CPL), is a useful spectroscopic technique for structural studies of biological systems composed of chiral molecules. The present study evaluated the effects of CPL on germination, hypocotyl elongation and biomass production of Arabidopsis and lettuce. Higher germination rates were observed when Arabidopsis and lettuce seedlings were irradiated with red right-handed CPL (R-CPL) than with red left-handed CPL (L-CPL). Hypocotyl elongation was effectively inhibited when Arabidopsis and lettuce seedlings were irradiated with red R-CPL than with red L-CPL. This difference was not observed when a phytochrome B (phyB) deficient mutant of Arabidopsis was irradiated, suggesting that inhibition of elongation by red R-CPL was mediated by phyB. White R-CPL induced greater biomass production by adult Arabidopsis plants, as determined by their fresh shoot weight, than white L-CPL. To determine the molecular basis of these CPL effects, CD spectra and the effect of CPL on the photoreaction of a sensory module of Arabidopsis phyB were measured. The red light-absorbing form of phyB showed a negative CD in the red light-absorbing region, consistent with the results of germination, inhibition of hypocotyl elongation and biomass production. L-CPL and R-CPL, however, did not differ in their ability to induce the interconversion of the red light-absorbing and far-red light-absorbing forms of phyB. These findings suggest that these CPL effects involve phyB, along with other photoreceptors and the photosynthetic process.

Inter- and intraspecific variation of spider mite susceptibility to fungal infections: Implications for the long-term success of biological control.

Spider mites are severe pests of several annual and perennial crops worldwide, often causing important economic damages. As rapid evolution of pesticide resistance in this group hampers the efficiency of chemical control, alternative control strategies, such as the use of entomopathogenic fungi, are being developed. However, while several studies have focused on the evaluation of the control potential of different fungal species and/or isolates as well as their compatibility with other control methods (e.g., predators or chemical pesticides), knowledge on the extent of inter‐ and intraspecific variation in spider mite susceptibility to fungal infection is as yet incipient. Here, we measured the mortality induced by two generalist fungi, Beauveria bassiana and Metarhizium brunneum, in 12 spider mite populations belonging to different Tetranychus species: T. evansi, T. ludeni, and T. urticae (green and red form), within a full factorial experiment. We found that spider mite species differed in their susceptibility to infection by both fungal species. Moreover, we also found important intraspecific variation for this trait. These results draw caution on the development of single strains as biocontrol agents. Indeed, the high level of intraspecific variation suggests that (a) the one‐size‐fits‐all strategy may fail to control spider mite populations and (b) hosts resistance to infection may evolve at a rapid pace. Finally, we propose future directions to better understand this system and improve the long‐term success of spider mite control strategies based on entomopathogenic fungi.

Neuroprotective and hepatoprotective effect of whole red rice forms against oxidative stress in streptozotocin induced diabetic rats

Whole red rice ( Oryza sativa L.), consumed as staple food in many countries worldwide, is rich in phytochemicals, mainly antioxidants having health potency. The present study evaluated the neuro- and hepatoprotective efficacy of raw and parboiled whole red rice against streptozotocin (STZ) induced oxidative stress, and antioxidant metabolism both in diabetic and normal non-diabetic rats. Wistar rats were rendered diabetic by a single intraperitoneal ( i.p.) injection of streptozotocin (45 mg/kg body wt.), supplemented with raw and parboiled whole red rice for six weeks . Results revealed a drastic increase in oxidative stress markers, such as lipid peroxidation, nitric oxide (NO) level, hydroperoxide (HP) level; antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), thioredoxin reductases (TR), glutathione S-transferase ( GST), acetylcholinesterase (AChE) activity, and decrease in endogenous antioxidant, glutathione (GSH) in diabetic rat. On the other hand, addition of raw and parboiled whole red rice in the diet counteracted the STZ induced oxidative stress in diabetic groups and improved the neuro and hepatic antioxidant system. Both, parboiled and raw red rice forms, exercised similar impacts in diabetic as well as in normal rats suggesting improved antioxidant defence mechanism. Results have shown that the whole red rice possess antidiabetic potential with the antioxidant improving ability, and could be utilized as dietary supplements in diabetes management.

Engineering the photoactive orange carotenoid protein with redox-controllable structural dynamics and photoprotective function

Photosynthesis requires various photoprotective mechanisms for survival of organisms in high light. In cyanobacteria exposed to high light, the Orange Carotenoid Protein (OCP) is reversibly photoswitched from the orange (OCPO) to the red (OCPR) form, the latter binds to the antenna (phycobilisomes, PBs) and quenches its overexcitation. OCPR accumulation implicates restructuring of a compact dark-adapted OCPO state including detachment of the N-terminal extension (NTE) and separation of protein domains, which is reversed by interaction with the Fluorescence Recovery Protein (FRP). OCP phototransformation supposedly occurs via an intermediate characterized by an OCPR-like absorption spectrum and an OCPO-like protein structure, but the hierarchy of steps remains debatable. Here, we devise and analyze an OCP variant with the NTE trapped on the C-terminal domain (CTD) via an engineered disulfide bridge (OCPCC). NTE trapping preserves OCP photocycling within the compact protein structure but precludes functional interaction with PBs and especially FRP, which is completely restored upon reduction of the disulfide bridge. Non-interacting with the dark-adapted oxidized OCPCC, FRP binds reduced OCPCC nearly as efficiently as OCPO devoid of the NTE, suggesting that the low-affinity FRP binding to OCPO is realized via NTE displacement. The low efficiency of excitation energy transfer in complexes between PBs and oxidized OCPCC indicates that OCPCC binds to PBs in an orientation suboptimal for quenching PBs fluorescence. Our approach supports the presence of the OCPR-like intermediate in the OCP photocycle and shows effective uncoupling of spectral changes from functional OCP photoactivation, enabling redox control of its structural dynamics and function.

Binding of red form of Orange Carotenoid Protein (OCP) to phycobilisome is not sufficient for quenching

The Orange Carotenoid Protein (OCP) is responsible for photoprotection in many cyanobacteria. Absorption of blue light drives the conversion of the orange, inactive form (OCPO) to the red, active form (OCPR). Concomitantly, the N–terminal domain (NTD) and the C–terminal domain (CTD) of OCP separate, which ultimately leads to the formation of a quenched OCPR–PBS complex. The details of the photoactivation of OCP have been intensely researched. Binding site(s) of OCPR on the PBS core have also been proposed. However, the post–binding events of the OCPR–PBS complex remain unclear. Here, we demonstrate that PBS–bound OCPR is not sufficient as a PBS excitation energy quencher. Using site–directed mutagenesis, we generated a suite of single point mutations at OCP Leucine 51 (L51) of Synechocystis 6803. Steady–state and time–resolved fluorescence analyses demonstrated that all mutant proteins are unable to quench the PBS fluorescence, owing to either failed OCP binding to PBS, or, if bound, an OCP–PBS quenching state failed to form. The SDS–PAGE and Western blot analysis support that the L51A (Alanine) mutant binds to the PBS and therefore belongs to the second category. We hypothesize that upon binding to PBS, OCPR likely reorganizes and adopts a new conformational state (OCP3rd) different than either OCPO or OCPR to allow energy quenching, depending on the cross–talk between OCPR and its PBS core–binding counterpart.

Risk factors of self-interruption of medications for mental disorders in pregnancy

The current study examined factors associated with the increased risk of self-interruption of medications for mental disorders in pregnancy. We reviewed the obstetric records of all singleton deliveries beyond 22 weeks of gestation at Japanese Red Cross Katsushika Maternity Hospital form January 2014 through July 2018. We examined the clinical and social characteristics and obstetric outcomes in women with self-interruption of medications for mental disorders in pregnancy in comparison with those in women who continued the medications throughout the pregnancy. There were 177 women who were diagnosed with mental disorders requiring medications before pregnancy by Japanese psychiatric specialists. Of these, 46 (26%) self-interruptedi their medications during the first trimester of their pregnancies. The rates of partners with mental disorders (odds ratio: 4.39, p = 0.01) and presence of social support (odds ratio: 2.50, p = 0.02) in women showing self-interruption of their medications were significantly higher than those in women continuing the medications. In the presence of both factors of a partner's mental disorders and social support, the odds ratio for self-interruption of medications increased to 15.9 (95% confidence interval 2.4-100, p &lt; 0.01). We believe that it may be the possible to identify women at high risk of self-interruption of medications during pregnancy. In women requiring perinatal mental health care, mental health support of their partners may also be needed.

310P - Laser and PDT for the oral leukoplakia

BackgroundLeukoplakia is a oral pathology characterized by an increased risk for malignant transformation. Histopathology of leukoplakia can disclose hyperkeratosis with dysplasia or carcinoma or hyperkeratosis or parakeratosis without dysplasia. Existence of the different treatment options not decrease the the rates of recurrence up to 35%. MethodsTotally 40 patients with OL (homogenous type - 35 cases; verrucous type - 4 case; erythroleukoplakia - one case) were recruited for the study. In a period September 2017 – may 2019 all the patients were screened, and only those patients with clinically, and histologically proven OL was included under the study. All the patients had the history of using tobacco in smoking or smokeless form (Nasvay). 1470nm diode laser ablation was done under 5- ALA induced spectroscopy. In case of detection a pathological fluorescence after the session treatment was accompanied with PDT treatment. 10% ALA emulsion (0.1g of 99.9% ALA powder in 1 ml liquid paraffin) was used freshly just before the start of the therapy to avoid deterioration of the emulsion on contact with light. Diode lamp (Vostok -1, Uzbekistan) with power - 5 W, wavelength -630 ± 5nm was used as the light source. Total dose delivered per session was 100 J/cm 2. The therapy was performed in a one session. All patients were recalled at 4 weeks after completion of the last session of modified therapy. Lesion response was characterized as follows: Complete response (CR), ; partial response (PR), reduction at least 20% in diameter; no response (NR). The patients were followed-up for a period of 1 year. ResultsOne month (4 weeks) after 1470nm diode laser ablation + ALA-PDT, the response was evaluated based on clinical examination. CR was obtained in 35 out of 40 patients; PR in 5 out of 40 patients; and NR no patient. During the follow-up, no changes in the clinical appearance of the treated lesions were recorded. There was no recurrence of the lesion noted in CR cases. During the incubation period of 3 h after ALA application, the patients did not have any adverse effect in the form of pain, redness or discomfort. Patients did not have any other side effects. ConclusionsOur observations suggest that topical ALA mediated PDT with laser ablation is a promising means of treating OL. Legal entity responsible for the studyTashkent medical academy. FundingHas not received any funding. DisclosureThe author has declared no conflicts of interest.