ABSTRACT
Hyperpolarized 13C-labeled fumarate probes tissue necrosis via the production of 13C-malate. Despite its promises in detecting tumor necrosis and kidney injuries, its clinical translation has been limited, primarily due to the low solubility in conventional glassing solvents. In this study, we introduce a new formulation of fumarate for dissolution dynamic nuclear polarization (DNP) by using meglumine as a counterion, a nonmetabolizable derivative of sorbitol. We have found that meglumine fumarate vitrifies by itself with enhanced water solubility (4.8 M), which is expected to overcome the solubility-restricted maximum concentration of hyperpolarized fumarate after dissolution. The achievable liquid-state polarization level of meglumine-fumarate is more than doubled (29.4 ± 1.3%) as compared to conventional dimethyl sulfoxide (DMSO)-mixed fumarate (13.5 ± 2.4%). In vivo comparison of DMSO- and meglumine-prepared 50-mM hyperpolarized [1,4-13C2]fumarate shows that the signal sensitivity in rat kidneys increases by 10-fold. As a result, [1,4-13C2]aspartate and [13C]bicarbonate in addition to [1,4-13C2]malate can be detected in healthy rat kidneys in vivo using hyperpolarized meglumine [1,4-13C2]fumarate. In particular, the appearance of [13C]bicarbonate indicates that hyperpolarized meglumine [1,4-13C2]fumarate can be used to investigate phosphoenolpyruvate carboxykinase, a key regulatory enzyme in gluconeogenesis.
Subject(s)
Carbon Isotopes , Fumarates , Kidney , Solubility , Animals , Fumarates/chemistry , Fumarates/metabolism , Rats , Kidney/metabolism , Carbon Isotopes/chemistry , Gluconeogenesis , Male , Rats, Sprague-DawleyABSTRACT
The feasibility of hyperpolarized [2-13C, 3-2H3]pyruvate for probing gluconeogenesis in vivo was investigated in this study. Whereas hyperpolarized [1-13C]pyruvate has clear access to metabolic pathways that convert pyruvate to lactate, alanine, and bicarbonate, its utility for assessing pyruvate carboxylation and gluconeogenesis has been limited by technical challenges, including spectral overlap and an obscure enzymatic step that decarboxylates the labeled carbon. To achieve unambiguous detection of gluconeogenic products, the carbonyl carbon in pyruvate was labeled with 13C. To prolong the T1 relaxation time, [2-13C, 3-2H3]pyruvate was synthesized and dissolved with D2O after dynamic nuclear polarization. The T1 of [2-13C, 3-2H3]pyruvate in D2O could be improved by 76.9% (79.6 s at 1 T and 74.5 s at 3 T) as compared to [2-13C]pyruvate in water. Hyperpolarized [2-13C, 3-2H3]pyruvate with D2O dissolution was applied to rat livers in vivo under normal feeding and fasting conditions. A gluconeogenic product, [2-13C]phosphoenolpyruvate, was observed at 149.9 ppm from fasted rats only, highlighting the utility of [2-13C, 3-2H3]pyruvate in detecting key gluconeogenic enzyme activities such as pyruvate carboxylase and phosphoenolpyruvate carboxykinase in vivo.
Subject(s)
Gluconeogenesis , Liver , Pyruvic Acid , Animals , Liver/metabolism , Liver/chemistry , Pyruvic Acid/metabolism , Pyruvic Acid/chemistry , Rats , Male , Rats, Sprague-Dawley , Carbon Isotopes/chemistryABSTRACT
The ERM protein family, which consists of three closely related proteins in vertebrates, ezrin, radixin, and moesin (ERM), is an ancient and important group of cytoplasmic actin-binding and organizing proteins. With their FERM domain, ERMs bind various transmembrane proteins and anchor them to the actin cortex through their C-terminal F-actin binding domain, thus they are major regulators of actin dynamics in the cell. ERMs participate in many fundamental cellular processes, such as phagocytosis, microvilli formation, T-cell activation and tumor metastasis. We have previously shown that, besides its cytoplasmic activities, the single ERM protein of Drosophila melanogaster, moesin, is also present in the cell nucleus, where it participates in gene expression and mRNA export. Here we study the mechanism by which moesin enters the nucleus. We show that the nuclear import of moesin is an NLS-mediated, active process. The nuclear localization sequence of the moesin protein is an evolutionarily highly conserved, conventional bipartite motif located on the surface of the FERM domain. Our experiments also reveal that the nuclear import of moesin does not require PIP2 binding or protein activation, and occurs in monomeric form. We propose, that the balance between the phosphorylated and non-phosphorylated protein pools determines the degree of nuclear import of moesin.
ABSTRACT
Single-stranded DNA binding proteins (SSBs) are ubiquitous across all domains of life and play essential roles via stabilizing and protecting single-stranded (ss) DNA as well as organizing multiprotein complexes during DNA replication, recombination, and repair. Two mammalian SSB paralogs (hSSB1 and hSSB2 in humans) were recently identified and shown to be involved in various genome maintenance processes. Following our recent discovery of the liquid-liquid phase separation (LLPS) propensity of Escherichia coli (Ec) SSB, here we show that hSSB2 also forms LLPS condensates under physiologically relevant ionic conditions. Similar to that seen for EcSSB, we demonstrate the essential contribution of hSSB2's C-terminal intrinsically disordered region (IDR) to condensate formation, and the selective enrichment of various genome metabolic proteins in hSSB2 condensates. However, in contrast to EcSSB-driven LLPS that is inhibited by ssDNA binding, hSSB2 phase separation requires single-stranded nucleic acid binding, and is especially facilitated by ssDNA. Our results reveal an evolutionarily conserved role for SSB-mediated LLPS in the spatiotemporal organization of genome maintenance complexes. At the same time, differential LLPS features of EcSSB and hSSB2 point to functional adaptations to prokaryotic versus eukaryotic genome metabolic contexts.
Subject(s)
DNA , Phase Separation , Animals , Humans , DNA-Binding Proteins/chemistry , DNA Repair , DNA Replication , DNA, Single-Stranded/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Mammals/geneticsABSTRACT
Superoxide, an anionic dioxygen molecule, plays a crucial role in redox regulation within the body but is implicated in various pathological conditions when produced excessively. Efforts to develop superoxide detection strategies have led to the exploration of organic-based contrast agents for magnetic resonance imaging (MRI). This study compares the effectiveness of two such agents, nTMV-TEMPO and kTMV-TEMPO, for detecting superoxide in a mouse liver model with lipopolysaccharide (LPS)-induced inflammation. The study demonstrates that kTMV-TEMPO, with a strategically positioned lysine residue for TEMPO attachment, outperforms nTMV-TEMPO as an MRI contrast agent. The enhanced sensitivity of kTMV-TEMPO is attributed to its more exposed TEMPO attachment site, facilitating stronger interactions with water protons and superoxide radicals. EPR kinetics experiments confirm kTMV-TEMPO's faster oxidation and reduction rates, making it a promising sensor for superoxide in inflamed liver tissue. In vivo experiments using healthy and LPS-induced inflamed mice reveal that reduced kTMV-TEMPO remains MRI-inactive in healthy mice but becomes MRI-active in inflamed livers. The contrast enhancement in inflamed livers is substantial, validating the potential of kTMV-TEMPO for detecting superoxide in vivo. This research underscores the importance of optimizing contrast agents for in vivo imaging applications. The enhanced sensitivity and biocompatibility of kTMV-TEMPO make it a promising candidate for further studies in the realm of medical imaging, particularly in the context of monitoring oxidative stress-related diseases.
Subject(s)
Superoxides , Tobacco Mosaic Virus , Mice , Animals , Contrast Media/chemistry , Lipopolysaccharides , Magnetic Resonance Imaging/methods , LiverABSTRACT
Protein adulteration is a common fraud in the food industry due to the high price of protein sources and their limited availability. Total nitrogen determination is the standard analytical technique for quality control, which is incapable of distinguishing between protein nitrogen and nitrogen from non-protein sources. Three benchtops and one handheld near-infrared spectrometer (NIRS) with different signal processing techniques (grating, Fourier transform, and MEM-micro-electro-mechanical system) were compared with detect adulteration in protein powders at low concentration levels. Whey, beef, and pea protein powders were mixed with a different combination and concentration of high nitrogen content compounds-namely melamine, urea, taurine, and glycine-resulting in a total of 819 samples. NIRS, combined with chemometric tools and various spectral preprocessing techniques, was used to predict adulterant concentrations, while the limit of detection (LOD) and limit of quantification (LOQ) were also assessed to further evaluate instrument performance. Out of all devices and measurement methods compared, the most accurate predictive models were built based on the dataset acquired with a grating benchtop spectrophotometer, reaching R2P values of 0.96 and proximating the 0.1% LOD for melamine and urea. Results imply the possibility of using NIRS combined with chemometrics as a generalized quality control tool for protein powders.
Subject(s)
Nitrogen , Spectroscopy, Near-Infrared , Animals , Cattle , Spectroscopy, Near-Infrared/methods , Powders , Whey , Urea , Food Contamination/analysisABSTRACT
Background and Objective: Primary dysmenorrhea (PD) is one of the most common clinical disorders in women of reproductive age. Our aim was to examine whether a twice-weekly thirty-minute Aviva exercise intervention could result in improvements in pain level and body awareness in patients with PD. Materials and Methods: In our prospective observational trial, the observation period included two consecutive menstrual cycles and the period of the next menstrual bleeding. The first menstrual bleeding period was the first measurement time (T1), the second was the second measurement time (T2), and the third was the third measurement time (T3) in a total of 78 volunteers. The primary endpoint was the change in the level of menstrual pain according to the Numeric Rating Scale (NRS) questionnaire between the intervention group (IG) and the control group (CG) at T1, T2, and T3. In this study, the secondary outcomes were the differences between the IG and CG regarding the different subscales of the Hungarian version of the Body Awareness Questionnaire (BAQ-H) at T1, T2, and T3; the Borg scale results of the IG; and adherence to the intervention. Statistical tests such as independent-sample t-tests, chi-square tests, Pearson's linear correlation coefficient, and repeated-measure ANCOVA were used for the analyses. Results: In total, 78 volunteers were enrolled: 40 persons in the IG and 38 in the CG. There was a significant change in the level of menstruation pain according to the NRS questionnaire between the IG and CG (p < 0.001). There was no significant difference between the IG and CG regarding the different subscales of the BAQ-H. Only in the case of the "Note responses or changes in body process" subscale of the BAQ-H was there a trend-like effect from the Aviva exercises (p = 0.086). Conclusions: The Aviva exercise could contribute to pain relief from PD. Regarding body awareness, no significant difference was found between the two groups. Due to the short detection period and prospective observational design, our results are preliminary and need to be confirmed in larger clinical trials.
Subject(s)
Dysmenorrhea , Exercise Therapy , Female , Humans , Dysmenorrhea/therapy , Exercise/physiology , Exercise Therapy/methods , Menstruation , Pain Management/methods , Prospective StudiesABSTRACT
The FERM domain is a conserved and widespread protein module that appeared in the common ancestor of amoebae, fungi, and animals, and is therefore now found in a wide variety of species. The primary function of the FERM domain is localizing to the plasma membrane through binding lipids and proteins of the membrane; thus, for a long time, FERM domain-containing proteins (FDCPs) were considered exclusively cytoskeletal. Although their role in the cytoplasm has been extensively studied, the recent discovery of the presence and importance of cytoskeletal proteins in the nucleus suggests that FDCPs might also play an important role in nuclear function. In this review, we collected data on their nuclear localization, transport, and possible functions, which are still scattered throughout the literature, with special regard to the role of the FERM domain in these processes. With this, we would like to draw attention to the exciting, new dimension of the role of FDCPs, their nuclear activity, which could be an interesting novel direction for future research.
Subject(s)
Cytoskeletal Proteins , FERM Domains , Animals , Protein Structure, Tertiary , Cytoskeletal Proteins/genetics , Cytoskeletal Proteins/metabolism , Cell Membrane/metabolism , Cell Nucleus/metabolismABSTRACT
The effect of Haematococcus pluvialis (HP) (0.25â¼1.25 %) as a colorant during high moisture extrusion (50 %) on the texture and microstructural properties of soy protein-based high moisture meat analogs (HMMA) was evaluated. Furthermore, the stability of HP-induced meat like color of the HMMA as a function of light exposure, freeze/thawing, frozen storage and cooking temperature and duration was investigated. The addition of HP reduced the elasticity of HMMA but enhanced its hardness, chewiness, and resilience. HP addition at low levels promoted the flexible and disordered regions within the protein secondary structure while excessive HP addition was unfavorable for protein cross-linking. The optimal degree of texturization was achieved with 0.75 % HP. Sensory evaluations revealed that HMMA with 1 %HP had a color similar to fresh beef sirloin, while HMMA with 0.25 % HP had a color closer to fresh pork loin. Light exposure induced the greatest color loss of the meat analogs compared with the cooking and frozen storage. The a* value of HMMA containing 1.25 % HP decreased by 30 % during the 14 days of light exposure. Frozen storage at darkness efficiently preserved the meat-like color of the extrudates. Overall, HP was found as promising colorant for HMMA production but the storage condition of the extrudates should be carefully optimized.
Subject(s)
Meat Substitutes , Meat , Animals , Cattle , Meat/analysis , Cooking , FreezingABSTRACT
BACKGROUND: Primary dysmenorrhea (PD) is one of the most common diseases in women of reproductive age. Our aim was to examine whether a twice-weekly thirty-minute exercise intervention could result in a difference in the pulsatility index (PI) of the uterine arteries (UAs) and level of menstrual pain in patients with PD. METHODS: In our prospective observational trial, the observation period included one spontaneous menstrual cycle and the consecutive time of the next menstruation of all participants, aged 18-44, with no extensive sports experience. In total, 73 volunteers were enrolled: 38 persons in the intervention group (IG) and 35 in the control group (CG). The intervention program was accompanied by music, performed in groups under the supervision of a qualified instructor in Hungary. The primary outcome was the difference between the IG and CG regarding the PI values of UAs at the 1st and the 2nd ultrasound (US) Doppler flowmetry. The secondary outcome was the difference between the IG and CG regarding the PI of UAs and menstrual pain measured by using the Numeric Rating Scale and adherence to the intervention. Statistical tests such as an independent-samples t-test, chi-square test, Mann-Whitney test and analysis of covariance (ANCOVA) were used during the analyses. RESULTS: Examining the mean of the PI of UAs in the IG and the CG at the 1st and the 2nd US measurement, a significant difference was found in the change in the measured value (Z = -2.545; p = 0.011). The IG showed a significantly higher increase in the mean of the PI of UAs (Median = 0.825) than the CG (Median = 0.130). The difference in the PI of the UAs of the IG and the CG is not related to the level of pain in any group (p = 0.336) and not related to the whole sample (p = 0.354); furthermore, the level of pain did not significantly differ between the two groups. CONCLUSIONS: Our study is the first to document the significant effects of mild-to-moderate exercise training on the change in the PI of the UAs in individuals with PD. The IG had a reduced blood flow due to circulatory redistribution after exercise. The level of menstrual pain of primary dysmenorrhea patients is independent of the level of blood circulation regarding the PI of the UAs. Randomized controlled studies with more participants and a longer research period are needed to confirm our findings regarding the association between regular exercise and the PI of UAs. The study was registered at clinicaltrials.gov: NCT04618172.
ABSTRACT
Parahydrogen-induced polarization (PHIP) followed by polarization transfer to 13 C is a rapidly developing technique for the generation of 13 C-hyperpolarized substrates. Chirality plays an essential role in living systems and differential metabolism of enantiomeric pairs of metabolic substrates is well documented. Inspired by asymmetric hydrogenation, here we report stereoPHIP, which involves the addition of parahydrogen to a prochiral substrate with a chiral catalyst followed by polarization transfer to 13 C spins. We demonstrate that parahydrogen could be rapidly added to the prochiral precursor to both enantiomers of lactic acid (D and L), with both the (R,R) and (S,S) enantiomers of a chiral rhodium(I) catalyst to afford highly 13 C-hyperpolarized (over 20 %) L- and D-lactate ester derivatives, respectively, with excellent stereoselectivity. We also show that the hyperpolarized 1 H signal decays obtained with the (R,R) and (S,S) catalysts were markedly different. StereoPHIP expands the scope of conventional PHIP to the production of 13 C hyperpolarized chiral substrates with high stereoselectivity.
ABSTRACT
Orthorhombic molybdenum trioxide (α-MoO3) is well known as a photocatalyst, adsorbent, and inhibitor during methyl orange photocatalytic degradation via TiO2. Therefore, besides the latter, other active photocatalysts, such as AgBr, ZnO, BiOI, and Cu2O, were assessed via the degradation of methyl orange and phenol in the presence of α-MoO3 using UV-A- and visible-light irradiation. Even though α-MoO3 could be used as a visible-light-driven photocatalyst, our results demonstrated that its presence in the reaction medium strongly inhibits the photocatalytic activity of TiO2, BiOI, Cu2O, and ZnO, while only the activity AgBr is not affected. Therefore, α-MoO3 might be an effective and stable inhibitor for photocatalytic processes to evaluate the newly explored photocatalysts. Quenching the photocatalytic reactions can offer information about the reaction mechanism. Moreover, the absence of photocatalytic inhibition suggests that besides photocatalytic processes, parallel reactions take place.
ABSTRACT
PURPOSE: The need to detect and quantify brain lactate accurately by MRS has stimulated the development of editing sequences based on J coupling effects. In J-difference editing of lactate, threonine can be co-edited and it contaminates lactate estimates due to the spectral proximity of the coupling partners of their methyl protons. We therefore implemented narrow-band editing 180° pulses (E180) in MEGA-PRESS acquisitions to resolve separately the 1.3-ppm resonances of lactate and threonine. METHODS: Two 45.3-ms rectangular E180 pulses, which had negligible effects 0.15-ppm away from the carrier frequency, were implemented in a MEGA-PRESS sequence with TE 139 ms. Three acquisitions were designed to selectively edit lactate and threonine, in which the E180 pulses were tuned to 4.1 ppm, 4.25 ppm, and a frequency far off resonance. Editing performance was validated with numerical analyses and acquisitions from phantoms. The narrow-band E180 MEGA and another MEGA-PRESS sequence with broad-band E180 pulses were evaluated in six healthy subjects. RESULTS: The 45.3-ms E180 MEGA offered a difference-edited lactate signal with lower intensity and reduced contamination from threonine compared to the broad-band E180 MEGA. The 45.3 ms E180 pulse had MEGA editing effects over a frequency range larger than seen in the singlet-resonance inversion profile. Lactate and threonine in healthy brain were both estimated to be 0.4 ± 0.1 mM, with reference to N-acetylaspartate at 12 mM. CONCLUSION: Narrow-band E180 MEGA editing minimizes threonine contamination of lactate spectra and may improve the ability to detect modest changes in lactate levels.
Subject(s)
Brain , Lactic Acid , Humans , Lactic Acid/analysis , Magnetic Resonance Spectroscopy , Brain/diagnostic imaging , Phantoms, Imaging , ThreonineABSTRACT
Hyperpolarized (HP) NMR can improve the sensitivity of conventional NMR experiments by several orders of magnitude, thereby making it feasible to detect the signal of low sensitivity nuclei such as 13C and 15N nuclei in vivo. Hyperpolarized substrates are usually administered by direct injection into the bloodstream, and interaction with serum albumin can cause rapid decay of the hyperpolarized signal due to the shortening of the spin-lattice (T1) relaxation time. Here we report that the 15N T1 of 15N labeled, partially deuterated tris(2-pyridylmethyl)amine decreases dramatically upon binding to albumin to such an extent that no HP-15 signal could be detected. We also demonstrate that the signal could be restored using a competitive displacer, iophenoxic acid, which binds stronger to albumin than tris(2-pyridylmethyl)amine. The methodology presented here eliminates the undesirable effect of albumin binding and should widen the range of hyperpolarized probes for in vivo studies.
ABSTRACT
It is necessary to develop and deploy novel protein production to allow the establishment of a sustainable supply for both humans and animals, given the ongoing expansion of protein demand to meet the future needs of the increased world population and high living standards. In addition to plant seeds, green biomass from dedicated crops or green agricultural waste is also available as an alternative source to fulfill the protein and nutrient needs of humans and animals. The development of extraction and precipitation methods (such as microwave coagulation) for chloroplast and cytoplasmic proteins, which constitute the bulk of leaf protein, will allow the production of leaf protein concentrates (LPC) and protein isolates (LPI). Obtained LPC serves as a sustainable alternative source of animal-based protein besides being an important source of many vital phytochemicals, including vitamins and substances with nutritional and pharmacological effects. Along with it, the production of LPC, directly or indirectly, supports sustainability and circular economy concepts. However, the quantity and quality of LPC largely depend on several factors, including plant species, extraction and precipitation techniques, harvest time, and growing season. This paper provides an overview of the history of green biomass-derived protein from the early green fodder mill concept by Károly Ereky to the state-of-art of green-based protein utilization. It highlights potential approaches for enhancing LPC production, including dedicated plant species, associated extraction methods, selection of optimal technologies, and best combination approaches for improving leaf protein isolation.
ABSTRACT
Fresh-cut leafy vegetables are one of the most perishable products because they readily deteriorate in quality even during cold storage and have a relatively short shelf life. Since these products are in high demand, methods for rigorous quality control and estimation of freshness that are rapid and non-destructive would be highly desirable. The objective of the present research was to develop a rapid, non-destructive near-infrared spectroscopy (NIRS)-based method for the evaluation of changes during cold storage of lettuce using an aquaphotomics approach to monitor the water molecular structure in lettuce leaves. The reference measurements showed that after 6 days of dark, cold storage, the weight and water activity of lettuce leaves decreased and ß-carotene decreased, while chlorophylls slightly increased. Aquaphotomics characterization showed large differences in the lettuce leaves' spectra depending on their growth zone. Difference spectra, principal component analysis (PCA) and linear discriminant analysis (LDA) confirmed the differences in the inner and outer leaves and revealed that spectra change as a function of storage time. Partial least squares regression (PLSR) allowed the prediction of the time spent in storage with a coefficient of determination of R2 = 0.80 and standard error of RMSE = 0.77 days for inner, and R2 = 0.86 and RMSE = 0.66 days for outer leaves, respectively. The following water absorbance bands were found to provide the most information in the spectra: 1348, 1360, 1373, 1385, 1391, 1410, 1416, 1422, 1441, 1447, 1453, 1466, 1472, 1490, 1503, 1515, 1521, 1534 and 1571 nm. They were further used as water matrix coordinates (WAMACs) to define the water spectral patterns (WASPs) of lettuce leaves. The WASPs of leaves served to succinctly describe the state of lettuces during storage. The changes in WASPs during storage reveled moisture loss, damage to cell walls and expulsion of intracellular water, as well as loss of free and weakly hydrogen-bonded water, all leading to a loss of juiciness. The WASPs also showed that damage stimulated the defense mechanisms and production of vitamin C. The leaves at the end of the storage period were characterized by water strongly bound to collapsed structural elements of leaf tissues, mainly cellulose, leading to a loss of firmness that was more pronounced in the outer leaves. All of this information was reflected in the changes of absorbance in the identified WAMACs, showing that the water molecular structure of lettuce leaves accurately reflects the state of the lettuce during storage and that WASPs can be used as a multidimensional biomarker to monitor changes during storage.
ABSTRACT
Determining and applying 'good' postharvest and quality control practices for otherwise highly sensitive fruits, such as sour cherry, is critical, as they serve as excellent media for a wide variety of microbial contaminants. The objective of this research was to report two series of experiments on the modified atmosphere storage (MAP) of sour cherries (Prunus cerasus L. var. Kántorjánosi, Újfehértói fürtös). Firstly, the significant effect of different washing pre-treatments on various quality indices was examined (i.e., headspace gas composition, weight loss, decay rate, color, firmness, soluble solid content, total plate count) in MAP-packed fruits. Subsequently, the applicability of near infrared (NIR) spectroscopy combined with chemometrics was investigated to detect the effect of various storage conditions (packed as control or MAP, stored at 3 or 5 °C) on sour cherries of different perceived ripeness. Significant differences were found for oxygen concentration when two perforations were applied on the packages of 'Kántorjánosi' (p < 0.01); weight loss when 'Kánorjánosi' (p < 0.001) and 'Újfehértói fürtös' (p < 0.01) were packed in MAP; SSC when 'Újfehértói fürtös' samples were ozone-treated (p < 0.05); and total plate count when 'Kántorjánosi' samples were ozone-treated (p < 0.01). The difference spectra reflected the high variability in the samples, and the detectable effects of different packaging. Based on the investigations with the soft independent modelling of class analogies (SIMCA), different packaging and storage resulted in significant differences in most of the cases even on the first storage day, which in many cases increased by the end of storage. The soft independent modelling of class analogies proved to be suitable for classification with apparent error rates between 0 and 0.5 during prediction regardless of ripeness. The research findings suggest the further correlation of NIR spectroscopic and reference parameters to support postharvest handling and fast quality control.
Subject(s)
Ozone , Prunus avium , Prunus avium/chemistry , Spectroscopy, Near-Infrared , Fruit/chemistry , Ozone/analysis , AtmosphereABSTRACT
Transposons are genetic elements that use various mechanisms of transposition to move around the genome, thus posing a risk to genomic integrity. Repression of transposable elements (TEs) involves the complex PIWI pathway and several proteins associated with heterochromatinization. All players of TE repression are indispensable for proper reproductive fitness, as loss-of-function mutations in these genes result primarily in sterility and impaired reproductive development. When investigating the function of novel genes with similar phenotypes, elevated transposon expression in reproductive tissues can be a marker for involvement in the aforementioned processes. Here, we present a protocol for investigating TE levels in adult Drosophila ovaries, from dissection to data analysis.