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Chronic pain represents a significant unmet medical need, affecting one-fifth of the U.S. population. EC5026 is a small molecule inhibitor of the enzyme soluble epoxide hydrolase (sEH) which is being developed as a novel non-opioid, non-NSAID analgesic. EC5026 prolongs the action of epoxy fatty acids, endogenous analgesic lipid mediators that are rapidly metabolized by sEH. We evaluated the safety and pharmacokinetic profile of EC5026 in two phase I trials, a single-ascending dose (SAD) study and a fed-fasted study. The SAD study evaluated EC5026 doses ranging from 0.5 to 24 mg in healthy volunteers. EC5026 was well tolerated. No treatment-emergent adverse events were considered related to EC5026. No apparent treatment- or dose-related trends in laboratory results, vital signs, physical examinations, or electrocardiograms were observed. A linear, near-dose-proportional increase in exposure was observed with progressive doses in the SAD study; plasma exposure was below or near the lower limit of quantification after 0.5-2 mg doses. Mean half-lives ranged from 41.8 to 59.1 h. for doses of 8-24 mg, supporting a once-daily dosing regimen. In the fed-fasted study using 8 mg EC5026 tablets, higher peak concentrations (66%) and total exposures (53%) were observed under the fed condition. Plasma concentrations declined in a monoexponential manner with mean half-lives of 59.5 h. in the fed state and 66.9 h. in the fasted state. Future clinical trials using EC5026 for the treatment of pain are justified based on the favorable outcomes from both clinical trials along with preclinical evidence of analgesic activity.

Hydroxylamine nitrate (HAN) is a new type of high-energy oxidant used in controllable propulsion. The present study explores the reaction mechanism of HAN using density functional theory and constructs reaction mechanism diagrams to understand the mechanism of small-molecule gas formation during the thermal decomposition of HAN. Based on transition state theory, the half-life of each reaction is calculated under standard conditions and the kinetic parameters of each reaction are scanned across the temperature range 298.15–1200 K. The research revealed a vast half-life time scale forreaction of NO generation, NO 2 generation, NO+NO 2 generation, N 2 O generation and N 2 generation process, meaning thatthese reaction can not occur. The half-life of RDS5 is very short. However, the reaction is also limited by the concentration of the reactant HNO. However, increasing the temperature rapidly decreases the reaction half-life and the reaction can easily proceed. Taking 300 s as the easily reactive boundary point, the cut-off points of the rate-determining steps of Processes 1 (NO generation), 2 (NO 2 generation), 3 (NO + NO 2 generation), 4 and 5 (N 2 O generation) and 6 (N 2 generation) are 466 K, 468 K, 576 K, 587 K and 402 K, respectively. The calculated reaction mechanism revealed two isomeric transformations of HNO 2 and H 2 N 2 O 2 and three isomeric transformations of H 2 N 2 O. Both HNO 2 and H 2 N 2 O 2 are transformed via H-atom transfer, while H 2 N 2 O is transformed either by H-atom transfer or intramolecular rotation. As the formation reaction of NO 2 does not have the lowest free energy, the NO 2 product is easily converted to other products. When NO 2 coexists with NO, it is also easily converted to the stable products N 2 O, N 2 and NH 3. [ABSTRACT FROM AUTHOR]

This paper is devoted to the calculations of alpha decay half-lives of 1 8 6 − 2 1 8 Po (Z = 8 4) isotopes and actinide nuclei between the atomic numbers Z = 90 and Z = 102, using an energy-dependent potential model. The α -nucleus effective potential employed in this study includes a Coulomb potential, a centrifugal barrier potential, and a nuclear potential. The nuclear potential is chosen to be the energy-dependent singularized harmonic potential, typical of nuclear interactions. The role of the energy-dependent potential is very significant in this study. Indeed, when compared to the experimental data, our results are found to be improved when energy-dependent interaction potential is employed compared to when energy-independent interactions are used, especially for small values of the parameter γ as evidenced by the values of the root mean square deviation. [ABSTRACT FROM AUTHOR]

Summary: This study investigated the impact of heating temperatures (70 °C to 90 °C for up to 14 h) on lycopene and β‐carotene degradation kinetics in gac aril paste. Carotenoids extracted via low‐volume hexane‐assisted ultrasonic extraction revealed a first‐order degradation pattern with high correlation coefficients (0.96–0.97). Lycopene degraded faster at 90 °C (k = 12.05 × 102 h−1) than at 70 °C (5.41 × 102 h−1), while β‐carotene displayed relatively slower degradation. Corresponding half‐life (t1/2) values for lycopene ranged from 5.75 to 12.81 h, while β‐carotene displayed values of 9.48–20.2 h. β‐carotene showed superior thermal stability compared to lycopene across all temperatures. Additionally, colour changes indicated decreased brightness with increasing temperature. The DPPH radical scavenging activity and FRAP values decreased during gac fruit heating. Overall, this study emphasises temperature‐induced declines in lycopene and β‐carotene levels in gac aril paste and offers insights into their degradation kinetics under thermal processing. [ABSTRACT FROM AUTHOR]

In the tea-planting process, insecticides are commonly combined, potentially prolonging the pre-harvest interval and heightening the risk of dietary exposure. This study focused on three frequently used insecticides in tea cultivation: thiamethoxam, bifenthrin, and dinotefuran, aiming to investigate their dissipation behaviors and associated dietary risks upon individual and simultaneous application. The dissipation kinetics of thiamethoxam, bifenthrin, and dinotefuran were successfully characterized by first-order kinetics, yielding respective half-lives of 5.44, 9.81, and 10.16 days. Upon joint application, the dissipation half-lives of thiamethoxam and bifenthrin were notably prolonged compared with their individual applications, resulting in final concentrations after 28 days that were correspondingly elevated by 1.41 and 1.29 times. Assessment of the dietary intake risk revealed that the chronic and acute risk quotients associated with thiamethoxam and bifenthrin escalated by 1.44–1.59 times following their combined application. Although dietary risks associated with Tianmuhu white tea, as determined by the exposure assessment model, were deemed acceptable, the cumulative risks stemming from pesticide mixtures across various dietary sources warrant attention. Molecular docking analyses further unveiled that thiamethoxam and bifenthrin competitively bound to glutathione S-transferase (GST) at amino acid residues, notably at the 76th GLU and the 25th PHE, pivotal in the metabolism and absorption of exogenous substances. Moreover, the interactions between P-glycoprotein and pesticides during transport and absorption were likely to influence dissipation behaviors post-joint application. This research offers valuable insights and data support for optimizing joint pesticide application strategies and assessing risks associated with typical pesticides used in tea cultivation. [ABSTRACT FROM AUTHOR]

An analytic solution is presented for the simultaneous substrate elimination problem that combines Michaelis-Menten (MM) consumption with an irreversible homo-dimerization process. The implicit solution involves logarithm and inverse tangent functions and perfectly agrees with the numerical solution of the differential equation. A solution is also presented for the generalized dynamical problem that simultaneously combines MM kinetics with first and second-order processes. The exact expressions for the half-life and the area under the curve are also presented for these problems. [ABSTRACT FROM AUTHOR]

Hydroxylamine nitrate (HAN) is a new type of high-energy oxidant used in controllable propulsion. The present study explores the reaction mechanism of HAN using density functional theory and constructs reaction mechanism diagrams to understand the mechanism of small-molecule gas formation during the thermal decomposition of HAN. Based on transition state theory, the half-life of each reaction is calculated under standard conditions and the kinetic parameters of each reaction are scanned across the temperature range 298.15–1200 K. The research revealed a vast half-life time scale forreaction of NO generation, NO2 generation, NO+NO2 generation, N2O generation and N2 generation process, meaning thatthese reaction can not occur. The half-life of RDS5 is very short. However, the reaction is also limited by the concentration of the reactant HNO. However, increasing the temperature rapidly decreases the reaction half-life and the reaction can easily proceed. Taking 300 s as the easily reactive boundary point, the cut-off points of the rate-determining steps of Processes 1 (NO generation), 2 (NO2 generation), 3 (NO + NO2 generation), 4 and 5 (N2O generation) and 6 (N2 generation) are 466 K, 468 K, 576 K, 587 K and 402 K, respectively. The calculated reaction mechanism revealed two isomeric transformations of HNO2 and H2N2O2 and three isomeric transformations of H2N2O. Both HNO2 and H2N2O2 are transformed via H-atom transfer, while H2N2O is transformed either by H-atom transfer or intramolecular rotation. As the formation reaction of NO2 does not have the lowest free energy, the NO2 product is easily converted to other products. When NO2 coexists with NO, it is also easily converted to the stable products N2O, N2 and NH3.

Decreased appetite is a common clinical problem in captive rhesus macaques (Macaca mulatta). Mirtazapine, a tetracyclic antidepressant originally developed for humans, has shown promise as a safe and effective promoter of weight gain and appetite in several veterinary species including rhesus and cynomolgus macaques. Although mirtazapine is available as oral formulations, transdermal delivery in macaques with reduced appetite would allow quick, painless, topical application. Here we describe the pharmacokinetics of a single application of a widely available veterinary transdermal mirtazapine formulation in 6 rhesus macaques. A dose of 0.5 mg/kg of transdermal mirtazapine ointment that has proven to be effective in rhesus was applied to the caudal pinnae of 3 female and 3 male young adult macaques. Serum was collected at 0, 0.5, 1, 3, 6, 8, 12, 24, 36, 48, and 72 h after administration. Our data indicate transdermal mirtazapine is absorbed at a lower level in rhesus as compared with published values in domestic cats (rhesus peak serum concentration: 1.2 ± 0.3 ng/mL), while drug half-life is longer than that reported in cats (rhesus: 33 ± 7 h). Mirtazapine reaches peak plasma concentrations in rhesus at 16 ± 10 h after administration; our model indicates that up to 5 d of serial dosing may be necessary to reach steady state. Our preliminary data also suggest that sex differences may contribute to efficacy and/or indicate sex-based differences, as male macaques reached Tmax more quickly than females (19 ± 2 h in females and 8 ± 3 h in males) and showed higher variation in half-life (33 ± 4 h in females and 34 ± 11 h in males). While previous work indicates clinical efficacy of the 0.5-mg/kg dosage in macaques, further investigation is warranted to determine if rhesus may benefit from higher recommended doses than companion animal species.

To examine the effect of organic amendments on the dissipation and nitrification inhibition activity of 2-amino 4-chloro 6-methyl pyrimidine (AM), a laboratory study was conducted on a sandy loam soil (Typic Hapludoll) receiving no organic amendments (Control) or application of farmyard manure (FYM) or pine needles biochar (PNB) @ 5 t ha−1. Dissipation of AM was initially faster during the first 7 d, and then it slowed down. Dissipation kinetics of AM conformed to the first-order model (R2 values = 0.973 to 0.981, significant at p ≤.01). The conventionally estimated time for the dissipation of half the amount of initially detected AM (half-life) was 15.9, 23.6, and 12.1 d for the control, FYM, and PNB treatments, respectively. All treatments receiving AM had numerically lower NO3−–N concentrations compared to no AM but significantly lower NO3−–N concentrations were recorded under, especially the PNB + AM treatment at 15, 30, 45, 60 and 90 d. The higher effectiveness of AM under PNB could be attributed to the sorption of AM in the micro-pores of PNB with slower release. Percent distribution of sorbed AM onto FYM and PNB revealed that desorption of sorbed AM from PNB was comparatively very limited. FTIR spectra revealed that both FYM and PNB strongly retained AM. Application of AM @1 mg kg−1 soil (equivalent to 2.24 kg ha−1) seems to effectively retard the nitrification of urea in soil amended with PNB and help reduce the leaching losses of NO3−–N in agriculture. [ABSTRACT FROM AUTHOR]

Abstract Background Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines are pivotal in combating coronavirus disease 2019 (COVID-19); however, the declining antibody titers postvaccination pose challenges for sustained protection and herd immunity. Although gut microbiome is reported to affect the early antibody response after vaccination, its impact on the longevity of vaccine-induced antibodies remains unexplored. Methods A prospective cohort study was conducted involving 44 healthy adults who received two doses of either the BNT162b2 or ChAdOx1 vaccine, followed by a BNT162b2 booster at six months. The gut microbiome was serially analyzed using 16S rRNA and shotgun sequencing, while humoral immune response was assessed using a SARS-CoV-2 spike protein immunoassay. Results Faecalibacterium prausnitzii was associated with robust and persistent antibody responses post-BNT162b2 vaccination. In comparison, Escherichia coli was associated with a slower antibody decay following ChAdOx1 vaccination. The booster immune response was correlated with metabolic pathways involving cellular functions and aromatic amino acid synthesis. Conclusions The findings of this study underscored the potential interaction between the gut microbiome and the longevity/boosting effect of antibodies following vaccination against SARS-CoV-2. The identification of specific microbial associations suggests the prospect of microbiome-based strategies for enhancing vaccine efficacy.

The Groundwater Ubiquity Score (GUS) is widely used to indicate the relative leachability of pesticides based on the soil half-life and the adsorption partition coefficient. In this manuscript, we derive mathematically the Theoretical Groundwater Ubiquity Score (TGUS) that, based on considerations of the preferential movement of pesticides to groundwater and a first-order pesticide degradation model, leads to a similar function as the GUS model. In the preferential flow model, movement to groundwater is fast, and the adsorption partition coefficient is thus not used for calculating the travel time to the groundwater (as it is in the advective-dispersive equation) but rather only determines the distribution of the pesticide between the water and soil phases. Both the GUS and TGUS models well predict the groundwater contamination of the originally studied pesticides for rainfall event(s) that caused pesticide leaching from 30 days after application. The theoretically derived Groundwater Ubiquity Score (TGUS) shows, in accordance with experimental evidence, that for leaching events shortly after spraying, the mass lost to (and resulting concentration in) groundwater is inversely related to the adsorption partition coefficient and not necessarily to the GUS index.

Abstract Exposure of firefighting instructors to polycyclic aromatic hydrocarbons (PAHs) such as naphthalene is unavoidable during live fire training. The study aimed to investigate naphthalene uptake by measuring the urinary excretion of the naphthalene metabolite 1,2-dihydroxynaphthalene (DHN), to describe the DHN elimination kinetics and to evaluate the results by comparison to further biomarkers of PAH exposure. N = 6 male non-smoking firefighting instructors completed five training sessions each in a residential fire simulation unit under respiratory protection. All participants provided two urine samples before and another seven samples within an 18-h-interval after each session. DHN was detected by gas chromatography/tandem mass spectrometry (GC–MS/MS) in all samples (n = 237) with median concentrations ranging from 3.3 µg/g crea. (range 0.9–10.2) before exposure to 134.2 µg/g crea. (43.4–380.4) post exposure. Maximum elimination found 3.3 h (median) after onset of exposure decreased with a mean half-life of 6.6 h to 27.1 µg/g crea. (15.7–139.5) 18 h after training. DHN sensitively indicated a presumed dermal naphthalene intake during training, showing similar elimination kinetics like other naphthalene metabolites. Internal exposure of the participants transiently exceeded exposures determined for non-smokers in the general population, but was lower than at other workplaces with PAH exposure. Despite limited uptake, accumulation is possible with daily exposure.

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines are pivotal in combating coronavirus disease 2019 (COVID-19); however, the declining antibody titers postvaccination pose challenges for sustained protection and herd immunity. Although gut microbiome is reported to affect the early antibody response after vaccination, its impact on the longevity of vaccine-induced antibodies remains unexplored. Methods: A prospective cohort study was conducted involving 44 healthy adults who received two doses of either the BNT162b2 or ChAdOx1 vaccine, followed by a BNT162b2 booster at six months. The gut microbiome was serially analyzed using 16S rRNA and shotgun sequencing, while humoral immune response was assessed using a SARS-CoV-2 spike protein immunoassay. Results: Faecalibacterium prausnitzii was associated with robust and persistent antibody responses post-BNT162b2 vaccination. In comparison, Escherichia coli was associated with a slower antibody decay following ChAdOx1 vaccination. The booster immune response was correlated with metabolic pathways involving cellular functions and aromatic amino acid synthesis. Conclusions: The findings of this study underscored the potential interaction between the gut microbiome and the longevity/boosting effect of antibodies following vaccination against SARS-CoV-2. The identification of specific microbial associations suggests the prospect of microbiome-based strategies for enhancing vaccine efficacy. [ABSTRACT FROM AUTHOR]

2,4-Dichlorophenoxyacetic acid (2,4-D) is an herbicide widely used in crops against broadleaf weeds. However, 2,4-D residues are considered an environmental pollutant in bodies of water. Phytoremediation with Plectranthus neochilus is a substantial strategy to remove 2,4-D from the aquatic environment. The objective of this study was to verify the efficiency of the association of the photostimulus by Light Emitting Diodes (LED) with P. neochilus to improve phytoremediation of 2,4-D in water. Phytoremediation was evaluated with the following samples: natural light, white LED, blue LED, and red LED, with and without the plant as controls. The data corresponding to the validation of the method were in accordance with the required parameters: R2: 0.9926; RSD: 1.74%; LOD: 0.075 mg.L−1; LOQ: 0.227 mg.L−1 and recovery by SPE was 76.57%. The efficiency of the association of LED with P. neochilus in the 28 days was: ambient light + plant (47.0%); white light + plant (37.10%); blue light + plant (26.80%); red light + plant (3.32%). This study demonstrated, for the first time, the efficiency of using LEDs light in association with P. neochilus for the phytoremediation of 2,4-D in water. NOVELTY STATEMENTS: Phytoremediation of organic compounds in water is a time-consuming process and generally unfavorable to the plant. This study demonstrated that the photostimulation with blue and red LED lights can accelerate the phytoremediation of the herbicide by P. neochilus, decreasing the t1/2 of 2,4-D in water by 2 and 5 times, respectively. We equate the time of this process to physical-chemical degradation methods, but without the use of reagents, creating a green strategy to accelerate the decontamination of water resources contaminated with pesticides. [ABSTRACT FROM AUTHOR]

Anthocyanins (ANCs) are water-soluble pigments that are useful as nutraceuticals due to their health benefits. This study was performed to evaluate the storage stability of purified and crude red grape ANCs in Raha Sweet (RS) during storage and to evaluate its sensory properties. ANCs were extracted from red grape pomace and purified with a macroporous resin. RS was prepared and colored with a synthetic food dye, Carmoisine (control), and ANCs (crude and purified). Pigments were extracted from RS weekly for a period of seven weeks and the absorbance was read spectrophotometrically. RS colored with ANCs was evaluated for its color and other sensory properties against another RS colored with the control. Results showed that the degradation of ANCs in RS followed the first-order reaction model, unlike the control, which showed no degradation during storage. The half-life of crude ANCs was three times higher than that of the purified ones, and RS colored with ANCs received a significantly (p < 0.05) lower score for color than that of RS colored with the control. ANCs could provide the food industry with a natural alternative to synthetic dyes to color foods with high sugar content that are stored for a short period of time. [ABSTRACT FROM AUTHOR]

An improved semi-empirical formula for cluster radioactivity half-lives is proposed by incorporating the influences of reduced mass, the blocking effect of unpaired nucleons and mass asymmetry on the basis of the original Tavares formula [Eur. Phys. J. A 49 (2013) 6]. Using this improved semi-empirical formula, the calculated results of the cluster radioactivity half-lives for the daughter nuclei around 2 0 8 Pb or its neighbors can reproduce the experimental data well with a corresponding root-mean-square deviation σ = 0.570, which proves the feasibility of this formula to calculate cluster radioactivity half-lives in trans-lead nuclei. The accuracy of the improved semi-empirical formula is improved by approximately 37% compared to its predecessor whose the root-mean-square deviation is 0.902. Meanwhile, the influence of deformation effect for cluster radioactivity is also briefly discussed. In addition, we extend this improved semi-empirical formula to predict the half-lives of 51 possible cluster radioactive candidates whose cluster radioactivity are energetically allowed or observed but not yet quantified in NUBASE2020. For comparison, some empirical and/or semi-empirical formulae are also used. [ABSTRACT FROM AUTHOR]

Exposure of firefighting instructors to polycyclic aromatic hydrocarbons (PAHs) such as naphthalene is unavoidable during live fire training. The study aimed to investigate naphthalene uptake by measuring the urinary excretion of the naphthalene metabolite 1,2-dihydroxynaphthalene (DHN), to describe the DHN elimination kinetics and to evaluate the results by comparison to further biomarkers of PAH exposure. N = 6 male non-smoking firefighting instructors completed five training sessions each in a residential fire simulation unit under respiratory protection. All participants provided two urine samples before and another seven samples within an 18-h-interval after each session. DHN was detected by gas chromatography/tandem mass spectrometry (GC–MS/MS) in all samples (n = 237) with median concentrations ranging from 3.3 µg/g crea. (range 0.9–10.2) before exposure to 134.2 µg/g crea. (43.4–380.4) post exposure. Maximum elimination found 3.3 h (median) after onset of exposure decreased with a mean half-life of 6.6 h to 27.1 µg/g crea. (15.7–139.5) 18 h after training. DHN sensitively indicated a presumed dermal naphthalene intake during training, showing similar elimination kinetics like other naphthalene metabolites. Internal exposure of the participants transiently exceeded exposures determined for non-smokers in the general population, but was lower than at other workplaces with PAH exposure. Despite limited uptake, accumulation is possible with daily exposure. [ABSTRACT FROM AUTHOR]

Peptide drugs are becoming star drug agents with high efficiency and selectivity which open up new therapeutic avenues for various diseases. However, the sensitivity to hydrolase and the relatively short half-life have severely hindered their development. In this study, a new generation artificial intelligence-based system for accurate prediction of peptide half-life was proposed, which realized the half-life prediction of both natural and modified peptides and successfully bridged the evaluation possibility between two important species (human, mouse) and two organs (blood, intestine). To achieve this, enzymatic cleavage descriptors were integrated with traditional peptide descriptors to construct a better representation. Then, robust models with accurate performance were established by comparing traditional machine learning and transfer learning, systematically. Results indicated that enzymatic cleavage features could certainly enhance model performance. The deep learning model integrating transfer learning significantly improved predictive accuracy, achieving remarkable R 2 values: 0.84 for natural peptides and 0.90 for modified peptides in human blood, 0.984 for natural peptides and 0.93 for modified peptides in mouse blood, and 0.94 for modified peptides in mouse intestine on the test set, respectively. These models not only successfully composed the above-mentioned system but also improved by approximately 15% in terms of correlation compared to related works. This study is expected to provide powerful solutions for peptide half-life evaluation and boost peptide drug development. [ABSTRACT FROM AUTHOR]

Cumulative evidence from several pre-clinical studies suggests that restoration of plasma DNase activity in a thrombo-inflammatory state may improve clinical outcomes. Following injury, hyperactivated immune cells release large amounts of granular proteins together with DNA, which often accumulate in the surrounding environment in so-called neutrophil extracellular traps (NETs). Degradation of excess NETs by systemic DNase administration offers a promising therapeutic approach to ameliorate inflammation and dissolve intravascular clots. In order to expand the therapeutic utility of human DNase I, a variant of the enzyme was developed that has both a prolonged systemic half-life and a higher catalytic activity compared to Dornase alfa (Pulmozyme®), the recombinant form of DNase I approved for inhaled therapy of cystic fibrosis. The hyperactive enzyme was "PASylated" by genetic fusion with a strongly hydrophilic and biodegradable PAS-polypeptide to increase its hydrodynamic volume and retard kidney filtration. A stable TurboCell™ CHO-K1-based cell line was generated which is suitable for the future production of PASylated DNase I according to good manufacturing practice (GMP). Furthermore, a robust bioprocess strategy was devised and an effective downstream process was developed. The final protein product is characterized by excellent purity, favorable physicochemical properties, a 14-fold higher DNA-degrading activity than Dornase alfa and a sustained pharmacokinetic profile, with a 22-fold slower clearance in rats. [ABSTRACT FROM AUTHOR]

The management of hemophilia A has undergone a remarkable revolution, in line with technological advancement. In the recent past, the primary concern associated with Factor VIII (FVIII) concentrates was the risk of infections, which is now almost resolved by advanced blood screening and viral inactivation methods. Improving patients' compliance with prophylaxis has become a key focus, as it can lead to improved health outcomes and reduced health care costs in the long term. Recent bioengineering research is directed toward prolonging the recombinant FVIII (rFVIII) coagulant activity and synthesising higher FVIII yields. As an outcome, B-domain deleted, polyethylene glycolated, single-chain, Fc-fused rFVIII, and rFVIIIFc-von Willebrand Factor-XTEN are available for patients. Moreover, emicizumab, a bispecific antibody, is commercially available, whereas fitusiran and tissue factor pathway inhibitor are in clinical trial stages as alternative strategies for patients with inhibitors. With these advancements, noninfectious complications, such as inhibitor development, allergic reactions, and thrombosis, are emerging concerns requiring careful management. In addition, the recent approval of gene therapy is a major milestone toward a permanent cure for hemophilia A. The vast array of treatment options at our disposal today empowers patients and providers alike, to tailor therapeutic regimens to the unique needs of each individual. Despite significant progress in modern treatment options, these highly effective therapies are markedly more expensive than conventional replacement therapy, limiting their access for patients in developing countries. [ABSTRACT FROM AUTHOR]

Phase-shift contrast agents consist of a liquid perfluorocarbon core that can be vaporized by ultrasound to generate echogenic contrast with excellent spatiotemporal control. The purpose of the present work was to evaluate the in vitro and in vivo behavior of condensed lipid-shelled nanodroplets (NDs) using different analytical procedures. Perfluorobutane NDs were prepared by condensation of precursor fluorescently labeled lipid-shelled microbubbles (MBs) and were characterized in terms of size distribution, gas core content and in vitro stability in blood, as well as for their acoustic vaporization behavior using a custom-made setup. In particular, the in vivo behavior of the NDs was thoroughly investigated after intravenous bolus injection in rats. To this end, we report, for the first time, the efficient use of three complementary detection procedures to assess the in vivo persistence of NDs: (i) ultrasound contrast imaging of vaporized NDs, (ii) gas chromatography–mass spectrometry to determine the perfluorobutane core content and (iii) fluorescence intensity measurement in the collected blood samples. The Coulter Counter Multisizer results confirmed the size distribution shift post-condensation. Furthermore, similar PFB concentrations from MB and ND suspensions were obtained, indicating an exceptionally low rate of MB breakage and spontaneous nanodroplet vaporization. As expected, these nanoscale droplets have longer circulation times compared with clinically approved MBs, and only slight variations in half-life were observed between the three monitoring procedures. Finally, echogenic signal observed in focal areas of the liver and spleen after vaporization was confirmed by accumulation of fluorescent nanodroplets in these organs. These results further contribute to our understanding of both the in vitro and in vivo behavior of sono-responsive nanodroplets, which is key to enabling efficient clinical translation. [ABSTRACT FROM AUTHOR]

We report on a new measurement of the half-life of 42Ar by following its decay over a time period of more than 13 years using γ-ray spectrometry. The obtained value of (39.5 ± 3.6) y confirms the only other, previously measured value of (33 ± 2) y. However, since partial outgassing of the accumulated Ar from the sample cannot be excluded, this value should be understood as a lower limit. The sample has now been stored in a gas-tight quartz ampoule to enable further measurements excluding the outgas effect in order to confirm our finding. [ABSTRACT FROM AUTHOR]

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This study aimed to investigate the dissipation pattern, risk assessment, and waiting period of myclobutanil on apple fruit (Malus domestica Borkh.) under temperate conditions in Kashmir, India. The study involved the application of myclobutanil 10 WP at a single recommended dosage (125 g a.i. ha−1) and double dosage (250 g a.i. ha−1) on Red Velox apple trees, 2 months before harvest. GC equipped with an electron capture detector was used to analyze myclobutanil residues in fruit samples. The study revealed that myclobutanil, at both recommended and double recommended doses, dissipated rapidly and became nondetectable after 55 and 60 days, respectively. The waiting period for myclobutanil application was determined to be 12.41 days for the single dose and 25.58 days for the double dose, respectively. These waiting periods were based on the maximum residue limit of 0.6 ppm as prescribed by the Codex Alimentarius Commission, Food Safety and Standards Authority of India, and European Commission. The study concludes that myclobutanil 10 WP is safe for consumers at both recommended and double recommended doses when applied 2 months before harvest. Risk assessment, considering the average daily apple consumption in India and theoretical maximum residue contributions (TMRCs), indicates negligible health hazards even at double the recommended dosage. The calculated TMRC values at Day 0 were significantly below the maximum permissible intake. For average and maximum myclobutanil residues at single and double doses, the TMRC values were found to be 0.0069 and 0.0070 mg day−1 person−1 and 0.0105 and 0.0106 mg day−1 person−1, respectively. These results indicate that myclobutanil, when used according to recommended dosages and waiting periods, poses minimal health risks to consumers. The study emphasizes the importance of prudent fungicide use to minimize fungicide residues on fruits, thereby ensuring their safety for consumption. [ABSTRACT FROM AUTHOR]

Understanding variant-specific differences in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral kinetics may explain differences in transmission efficiency and provide insights on pathogenesis and prevention. We evaluated SARS-CoV-2 kinetics from nasal swabs across multiple variants (Alpha, Delta, Epsilon, Gamma) in placebo recipients of the ACTIV-2/A5401 trial. Delta variant infection led to the highest maximum viral load and shortest time from symptom onset to viral load peak. There were no significant differences in time to viral clearance across the variants. Viral decline was biphasic with first- and second-phase decays having half-lives of 11 hours and 2.5 days, respectively, with differences among variants, especially in the second phase. These results suggest that while variant-specific differences in viral kinetics exist, post-peak viral load all variants appeared to be efficiently cleared by the host. Clinical Trials Registration. NCT04518410.

ABSTRACT Malaria rapid diagnostic tests (RDTs), which detect Plasmodium falciparum (Pf)-specific histidine-rich protein-2 (HRP2), have increasing importance for the diagnosis and control of malaria, especially also in regions where routine diagnosis by microscopy is not available. HRP2-based RDTs have a similar sensitivity to expert microscopy, but their reported low specificity can lead to high false positivity rates, particularly in high-endemic areas. Despite the widespread use of RDTs, models investigating the dynamics of HRP2 clearance following Pf treatment focus rather on short-term clearance of the protein. The goal of this observational cohort study was to determine the long-term kinetic of HRP2-levels in peripheral blood after treatment of uncomplicated malaria cases with Pf mono-infection using a 3-day course of artesunate/amodiaquine. HRP2 levels were quantified at enrollment and on days 1, 2, 3, 5, 7, 12, 17, 22, and 28 post-treatment initiation. The findings reveal an unexpectedly prolonged clearance of HRP2 after parasite clearance from capillary blood. Terminal HRP2 half-life was estimated to be 9 days after parasite clearance using a pharmacokinetic two-compartmental elimination model. These results provide evidence that HRP2 clearance has generally been underestimated, as the antigen remains detectable in capillary blood for up to 28 days following successful treatment, influencing RDT-based assessment following a malaria treatment for weeks. A better understanding of the HRP2 clearance dynamics is critical for guiding the diagnosis of malaria when relying on RDTs.IMPORTANCEDetecting Plasmodium falciparum, the parasite responsible for the severest form of malaria, typically involves microscopy, polymerase chain reaction (PCR), or rapid diagnostic tests (RDTs) targeting the histidine-rich protein 2 or 3 (HRP2/3). While microscopy and PCR quickly turn negative after the infection is cleared, HRP2 remains detectable for a prolonged period. The exact duration of HRP2 persistence had not been well defined. Our study in Gabon tracked HRP2 levels over 4 weeks, resulting in a new model for antigen clearance. We discovered that a two-compartment model accurately predicts HRP2 levels, revealing an initial rapid reduction followed by a much slower elimination phase that can take several weeks. These findings are crucial for interpreting RDT results, as lingering HRP2 can lead to false positives, impacting malaria diagnosis and treatment decisions.

BackgroundAutosomal dominant mutations in α-synuclein, TDP-43 and tau are thought to predispose to neurodegeneration by enhancing protein aggregation. While a subset of α-synuclein, TDP-43 and tau mutations has been shown to increase the structural propensity of these proteins toward self-association, rates of aggregation are also highly dependent on protein steady state concentrations, which are in large part regulated by their rates of lysosomal degradation. Previous studies have shown that lysosomal proteases operate precisely and not indiscriminately, cleaving their substrates at very specific linear amino acid sequences. With this knowledge, we hypothesized that certain coding mutations in α-synuclein, TDP-43 and tau may lead to increased protein steady state concentrations and eventual aggregation by an alternative mechanism, that is, through disrupting lysosomal protease cleavage recognition motifs and subsequently conferring protease resistance to these proteins.ResultsTo test this possibility, we first generated comprehensive proteolysis maps containing all of the potential lysosomal protease cleavage sites for α-synuclein, TDP-43 and tau. In silico analyses of these maps indicated that certain mutations would diminish cathepsin cleavage, a prediction we confirmed utilizing in vitro protease assays. We then validated these findings in cell models and induced neurons, demonstrating that mutant forms of α-synuclein, TDP-43 and tau are degraded less efficiently than wild type despite being imported into lysosomes at similar rates.ConclusionsTogether, this study provides evidence that pathogenic mutations in the N-terminal domain of α-synuclein (G51D, A53T), low complexity domain of TDP-43 (A315T, Q331K, M337V) and R1 and R2 domains of tau (K257T, N279K, S305N) directly impair their own lysosomal degradation, altering protein homeostasis and increasing cellular protein concentrations by extending the degradation half-lives of these proteins. These results also point to novel, shared, alternative mechanism by which different forms of neurodegeneration, including synucleinopathies, TDP-43 proteinopathies and tauopathies, may arise. Importantly, they also provide a roadmap for how the upregulation of particular lysosomal proteases could be targeted as potential therapeutics for human neurodegenerative disease.

Abstract Background Calcitonin gene-related peptide (CGRP) is the most promising candidate to become the first migraine biomarker. However, literature shows clashing results and suggests a methodological source for such discrepancies. We aimed to investigate some of these methodological factors to evaluate the actual role of CGRP as biomarker. Methods Previous to the experimental part, we performed a literature review of articles measuring CGRP in migraine patients. Using our 399 bio-bank sera samples, we performed a series of experiments to test the validity of different ELISA kits employed, time of sample processing, long-term storage, sampling in rest or after moderate exercise. Analysis of in-house data was performed to analyse average levels of the peptide and the effect of sex and age. Results Literature review shows the high variability in terms of study design, determination methods, results and conclusions obtained by studies including CGRP determinations in migraine patients. CGRP measurements depends on the method and specific kit employed, also on the isoform detected, showing completely different ranges of concentrations. Alpha-CGRP and beta-CGRP had median with IQR levels of 37.5 (28.2–54.4) and 4.6 (2.4–6.4)pg/mL, respectively. CGRP content is preserved in serum within the 24 first hours when samples are stored at 4°C after clotting and immediate centrifugation. Storages at -80°C of more than 6 months result in a decrease in CGRP levels. Moderate exercise prior to blood extraction does not modulate the concentration of the peptide. Age positively correlates with beta-CGRP content and men have higher alpha-CGRP levels than women. Conclusions We present valuable information for CGRP measurements in serum. ELISA kit suitability should be tested prior to the experiments. Alpha and beta-CGRP levels should be analysed separately as they can show different behaviours even within the same condition. Samples can be processed in a 24-h window if they have been kept in 4°C and should not be stored for more than 6 months at -80°C before assayed. Patients do not need to rest before the blood extraction unless they have performed a high-endurance exercise. For comparative studies, sex and age should be accounted for as these parameters can impact CGRP concentrations. Graphical Abstract

Introduction: Fungal diseases are frequently associated with elevated mortality rates in elasmobranchs. Currently, there is a notable absence of scientifically validated therapeutic medications that can ensure both effectiveness and safety when administered to this group of animals. The empirical prescription of azole antifungal agents, particularly voriconazole, has been posited as a potentially efficacious treatment approach for addressing most common mycoses in sharks and rays. However, there are still no published pharmacokinetic studies supporting its use in elasmobranchs and there is a lack of scientific base for its utilization in elasmobranchs. Methods: For this study, voriconazole was administered intravenously (IV) and intramuscularly (IM), at a single dose of 4 mg/kg to six adult undulate skates (Raja undulata). A washout period of 8 weeks was left between each route of administration. Blood samples were collected both before and at ten predetermined intervals after each dosing (0.25, 0.5, 1, 1.5, 2, 4, 8, 12, 24, and 36 h after drug administration). Plasma concentrations were quantified using a validated high-performance liquid chromatography method, and pharmacokinetic (PK) data was analyzed through non-compartmental methods. Results: The mean extrapolated concentration at 0 h (C0) after IV administration was 27.19 ± 7.15 µg/mL and the mean peak plasma concentrations (Cmax) ± SEM after IM administration resulted 2.98 ± 0.28 µg/mL at a mean time to maximum concentration (T max) of 1.33 ± 0.17 h. Terminal half-lives were calculated and resulted 11.18 ± 1.32 h for IV injections and 9.59 ± 1.38 h for IM injections. The area under the curve extrapolated to infinity was determined as 58.14 ± 2.79 h·µg/ml following IV injections and 37.60 ± 6.67 h·µg/ml following IM injections. The IM-administered voriconazole exhibited a mean absolute bioavailability of 64.67 ± 11.47%. Discussion: These discoveries provide backing for the possible application of voriconazole through the intramuscular route in undulate skates and support using lower dosage regimens compared to those required for oral administration, emphasizing the importance of conducting further pharmacokinetic studies with antifungals in elasmobranchs. [ABSTRACT FROM AUTHOR]

Using the properties of the Lambert function we review the analytical solutions of the Michaelis–Menten (MM) kinetics and other related models. We derive several quantities of interest such as the half-life and the area under the curve (AUC). The effect of varying the parameters in the Beal–Schnell–Mendoza solution and its asymptotic time behavior were analyzed. The Maclaurin expansion of the time evolution of substrate concentration up to sixth order is presented. These expressions were tested on the well-known problem of ethanol elimination from the human body and excellent agreement was found. In addition, a closed-form solution for the derived problem that combines simultaneously MM and zeroth-order kinetics is derived. This problem was solved by a suitable transformation of variables that casts the original differential equation into a functionally equivalent MM problem with termination time. To finish, analytical solutions for the MM process in parallel with zeroth- and first-order kinetics are presented here as well. We checked all equations against the numerically exact solution of the corresponding differential equation and perfect agreement was found in all cases. [ABSTRACT FROM AUTHOR]

Thiamethoxam, a systemic neonicotinoid insecticide, is widely accepted for use against various chewing and sucking insect pests. In this particular study, the authors examined the dissipation kinetics and metabolism of thiamethoxam in wheat using quick, easy, cheap, effective, rugged and safe (QuEChERS) methodology combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) over two years during the same cropping season. Residues of thiamethoxam and its metabolites in wheat plants were investigated following foliar spray of thiamethoxam at recommended dose (12.5 g a.i. ha−1) and four times the recommended dose (50.0 g a.i. ha−1) at earhead formation stage. Total residues of thiamethoxam and its metabolites in leaves at 0 day (2 h after spray) ranged from 3.69 to 14.36 mg kg−1 for the two doses over the two years. Clothianidin was the only metabolite detected in leaves at 0 day (2 h after spray); however at 7 days after spray nitroguanidine, N-desmethyl-thiamethoxam and 1-methyl-3-nitroguanidine were also detected. Total residues of thiamethoxam and its metabolites in soil at 0 day (2 h after spray) ranged from 0.23 to 0.84 mg kg−1. Thiamethoxam residue in leaves and soil followed first order kinetics with half-life ranging from 9.04 to 15.50 days for the two doses over the two study years. Residues of thiamethoxam and its metabolites in leaves, straw and grain at harvest time for both the doses were below their limit of quantification (0.01 mg kg−1). This information could be helpful for evaluating both regulatory and food safety decisions regarding use of neonicotinoids. [ABSTRACT FROM AUTHOR]

A critical parameter during the development of protein therapeutics is to endow them with suitable pharmacokinetic and pharmacodynamic properties. Small protein drugs are quickly eliminated by kidney filtration, and in vivo half-life extension is therefore often desired. Here, different half-life extension technologies were studied where PAS polypeptides (PAS300, PAS600), XTEN polypeptides (XTEN288, XTEN576), and an albumin binding domain (ABD) were compared for half-life extension of an anti-human epidermal growth factor receptor 2 (HER2) affibody-drug conjugate. The results showed that extension with the PAS or XTEN polypeptides or the addition of the ABD lowered the affinity for HER2 to some extent but did not negatively affect the cytotoxic potential. The half-lives in mice ranged from 7.3 h for the construct including PAS300 to 11.6 h for the construct including PAS600. The highest absolute tumor uptake was found for the construct including the ABD, which was 60 to 160% higher than the PASylated or XTENylated constructs, even though it did not have the longest half-life (9.0 h). A comparison of the tumor-to-normal-organ ratios showed the best overall performance of the ABD-fused construct. In conclusion, PASylation, XTENylation, and the addition of an ABD are viable strategies for half-life extension of affibody-drug conjugates, with the best performance observed for the construct including the ABD. [Display omitted] [ABSTRACT FROM AUTHOR]

Although the discovery of insulin 100 years ago revolutionized the treatment of diabetes, its therapeutic potential is compromised by its short half-life and narrow therapeutic index. Current long-acting insulin analogs, such as insulin-polymer conjugates, are mainly used to improve pharmacokinetics by reducing renal clearance. However, these conjugates are synthesized without sacrificing the bioactivity of insulin, thus retaining the narrow therapeutic index of native insulin, and exceeding the efficacious dose still leads to hypoglycemia. Here, we report a kind of di-PEGylated insulin that can simultaneously reduce renal clearance and receptor-mediated clearance. By impairing the binding affinity to the receptor and the activation of the receptor, di-PEGylated insulin not only further prolongs the half-life of insulin compared to classical mono-PEGylated insulin but most importantly, increases its maximum tolerated dose 10-fold. The target of long-term glycemic management in vivo has been achieved through improved pharmacokinetics and a high dose. This work represents an essential step towards long-acting insulin medication with superior safety in reducing hypoglycemic events. Distinct from conventional mono-PEGylated insulin, this work developed a kind of di-PEGylated insulin that could simultaneously reduce renal clearance and receptor-mediated clearance. [Display omitted] [ABSTRACT FROM AUTHOR]

Accelerating safety assessments for novel agrochemicals is imperative, advocating for in vitro setups to present pesticide biodegradation by soil microbiota before field studies. This approach enables metabolic profile generation in a controlled laboratory environment eliminating extrinsic factors. In the current study, ten different soil samples were utilized to check their capability to degrade Ametoctradin by their microbiota. Furthermore, five different fungal strains (Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, Lasiodiplodia theobromae, and Penicillium chrysogenum) were utilized to degrade Ametoctradin in aqueous media. A degradation pathway was established using the metabolic patterns created during the biodegradation of Ametoctradin. In contrast to 47% degradation (T1/2 of 34 days) when Ametoctradin was left in the soil samples, the fungal strain Aspergillus fumigatus demonstrated 71% degradation of parent Ametoctradin with a half-life (T1/2) of 16 days. In conclusion, soil rich in microorganisms effectively cleans Ametoctradin-contaminated areas while Fungi have also been shown to be an effective, affordable, and promising way to remove Ametoctradin from the environment. [ABSTRACT FROM AUTHOR]

Introduction: Consensus over the definition of recombinant factor VIII (rFVIII) product classification in haemophilia A is lacking. rFVIII products are often classified as standard half‐life (SHL) or extended half‐life (EHL); despite this, no universally accepted definition currently exists. One proposed definition includes half‐life, area under the curve, and technology designed to extend half‐life; however, the International Society on Thrombosis and Haemostasis defines activity over time as the most intuitive information for building treatment regimens and the World Federation of Hemophilia describes rFVIII product classification in terms of infusion frequency. Aim: To summarise published data on the clinical and pharmacokinetic criteria used to define rFVIII product classification. Methods: PubMed and EMBASE database searches of English‐language articles (2002–2022) were conducted using search strings to identify the relevant population, intervention, and outcomes (e.g., clinical and pharmacokinetic parameters). Articles then underwent title/abstract and full‐text screens. Results: Among 1147 identified articles, 62 were included. Half‐life was the most widely reported outcome with no clear trends or product groupings observed. No clear groupings emerged among other outcomes, including infusion frequency, consumption, and efficacy. As activity over time was reported in few articles, further investigation of its relevance to rFVIII product classification is warranted. Conclusion: The findings of this systematic literature review suggest that parameters other than half‐life might be important for the development of a comprehensive and clinically relevant rFVIII product classification definition. There seems to be an opportunity to consider parameters that are clinically meaningful and useful for shared decision‐making in haemophilia A treatment. [ABSTRACT FROM AUTHOR]

Chicken single‐chain fragment variable (IgY‐scFv) is a functional fragment and an emerging development in genetically engineered antibodies with a wide range of biomedical applications. However, scFvs have considerably shorter serum half‐life due to the absence of antibody Fc region compared with the full‐length antibody, and usually requires continuous intravenous administration for efficacy. A promising approach to overcome this limitation is to fuse scFv with immunoglobulin G (IgG) Fc region, for better recognition and mediation by the neonatal Fc receptor (FcRn) in the host. In this study, engineered mammalian ΔFc domains (CH2, CH3, and intact Fc region) were fused with anti‐canine parvovirus‐like particles avian IgY‐scFv to produce chimeric antibodies and expressed in the HEK293 cell expression system. The obtained scFv‐CH2, scFv‐CH3, and scFv‐Fc can bind with antigen specifically and dose‐dependently. Surface plasmon resonance investigation confirmed that scFv‐CH2, scFv‐CH3, and scFv‐Fc had different degrees of binding to FcRn, with scFv‐Fc showing the highest affinity. scFv‐Fc had a significantly longer half‐life in mice compared with the unfused scFv. The identified ΔFcs are promising for the development of engineered Fc‐based therapeutic antibodies and proteins with longer half‐lives. The avian IgY‐scFv‐mammalian IgG Fc region opens up new avenues for antibody engineering, and it is a novel strategy to enhance the rapid development and screening of functional antibodies in veterinary and human medicine. [ABSTRACT FROM AUTHOR]

Objectives: The aim of the present study was to assess the effect of gabapentin on blood pressure (BP) in cats with and without chronic kidney disease (CKD). Methods: A randomized, blinded, placebo-controlled crossover study was performed. A total of 29 cats were included: 13 cats with stable CKD (IRIS stage 2–4) and 16 apparently healthy cats (serum creatinine <1.6 mg/dl and urine specific gravity >1.035). The cats were evaluated twice, approximately 1 week apart, and BP (Doppler sphygmomanometry) was obtained 3 h after cats received either a single dose of gabapentin 10mg/kg PO or placebo. For each cat, BP readings were obtained at each visit using the same Doppler and sphygmomanometer unit, and the same cat holder and Doppler operator, in the same location. Results: After administration of a single dose of gabapentin (10 mg/kg PO), BP was significantly lower (median 122 mmHg, range 82–170) than after administration of the placebo (median 150 mmHg, range 102–191; P = 0.001). In the CKD subgroup, BP was significantly lower after administration of gabapentin (median 129 mmHg, range 96–170) than after administration of the placebo (median 155 mmHg, range 102–191; P = 0.008). In the healthy cat subgroup, BP was significantly lower after administration of gabapentin (median 121 mmHg, range 82–139) than after administration of the placebo (median 137 mmHg, range 102–177; P = 0.002). The median change in BP was −12 mmHg (range −95 to 10) for healthy cats and −12 mmHg (range −43 to 21) for cats with CKD (no significant difference between subgroups). Conclusions and relevance: Gabapentin may decrease arterial BP in cats with and without CKD and these findings should be taken into account when gabapentin is administered to patients in which measurement of BP is needed. [ABSTRACT FROM AUTHOR]

Apple, a major fruit of temperate Himalayas, is sprayed with chemical pesticides around 12 times during the cropping season. Various systemic and contact fungicides are applied for the management of major diseases. In order to manage disease, flusilazole 40 EC is frequently used. However, excessive chemical application has been found to be detrimental for consumer safety. Keeping in view consumer safety, risk assessment, the half‐life and waiting period for flusilazole 40 EC were evaluated on the Red Velox variety of apple. The QuEChERS (quick, easy, cheap, effective, rugged and safe) method and high‐performance liquid chromatography were adapted for sample processing and analysis, respectively. The recovery percentages of flusilazole at three fortification levels (0.04, 0.09 and 0.50 mg kg−1) were 98.85, 99.83 and 98.98%, respectively. Flusilazole at the recommended dose (80 g a.i. ha−1) left an initial deposit of 0.733 mg kg−1, which dissipated by 93.45% in 60 days and was non‐detectable beyond this period. Meanwhile flusilazole at double the recommended dose (160 g a.i. ha−1) left an initial deposit of 0.913 mg kg−1, which dissipated by 93.43% in 70 days and was non‐detectable beyond this period. Based on the maximum residue limit of 0.3 mg kg−1 as prescribed by the Codex Alimentarius Commission, a waiting period of 28.74 and 46.03 days was recorded for single and double doses, respectively. Moreover, in order to assess the consumer risk, theoretical maximum residue contributions (TMRCs) were derived using flusilazole residues (average and maximum) recorded at various time intervals and compared with the maximum permissible intake, which was found to be 0.42 mg per person per day. Based on the average per capita daily consumption of 6.76 g apple in India, the TMRC values were computed. Although the values of TMRC decreased below maximum permissible intake at the first day after application, indicating minimal consumer health risks, fruits sprayed with a double dose of flusilazole carried the risk even up to the tenth day after flusilazole application. The results of the present study will be valuable for safe and timely use of flusilazole on apple. [ABSTRACT FROM AUTHOR]

Background: Calcitonin gene-related peptide (CGRP) is the most promising candidate to become the first migraine biomarker. However, literature shows clashing results and suggests a methodological source for such discrepancies. We aimed to investigate some of these methodological factors to evaluate the actual role of CGRP as biomarker. Methods: Previous to the experimental part, we performed a literature review of articles measuring CGRP in migraine patients. Using our 399 bio-bank sera samples, we performed a series of experiments to test the validity of different ELISA kits employed, time of sample processing, long-term storage, sampling in rest or after moderate exercise. Analysis of in-house data was performed to analyse average levels of the peptide and the effect of sex and age. Results: Literature review shows the high variability in terms of study design, determination methods, results and conclusions obtained by studies including CGRP determinations in migraine patients. CGRP measurements depends on the method and specific kit employed, also on the isoform detected, showing completely different ranges of concentrations. Alpha-CGRP and beta-CGRP had median with IQR levels of 37.5 (28.2–54.4) and 4.6 (2.4–6.4)pg/mL, respectively. CGRP content is preserved in serum within the 24 first hours when samples are stored at 4°C after clotting and immediate centrifugation. Storages at -80°C of more than 6 months result in a decrease in CGRP levels. Moderate exercise prior to blood extraction does not modulate the concentration of the peptide. Age positively correlates with beta-CGRP content and men have higher alpha-CGRP levels than women. Conclusions: We present valuable information for CGRP measurements in serum. ELISA kit suitability should be tested prior to the experiments. Alpha and beta-CGRP levels should be analysed separately as they can show different behaviours even within the same condition. Samples can be processed in a 24-h window if they have been kept in 4°C and should not be stored for more than 6 months at -80°C before assayed. Patients do not need to rest before the blood extraction unless they have performed a high-endurance exercise. For comparative studies, sex and age should be accounted for as these parameters can impact CGRP concentrations. [ABSTRACT FROM AUTHOR]

This pilot study investigated the bioavailability and remineralization kinetics of the sediment-derived dissolved organic carbon (DOC) from the Gdańsk Deep, a depositional area in the Baltic Sea. This was assessed in the long-lasting (126 d) incubation experiment, in which the mixture of DOC from sediment pore water and bottom water was exposed to oxic microbial respiration with incubation of bottom water as a control run. The obtained decay curves allowed us to distinguish three DOC fractions: labile (DOCL), semi-labile (DOCSL), and refractory (DOCR). In bottom water, the refractory fraction was predominant and amounted to almost 85% of total DOC, whereas about 15% of DOC was bioavailable: 6% labile and 9% semi-labile. In contrast, DOC from pore water was much more bioavailable DOC (~55% of total DOC) and contained 11% DOCL and 44% DOCSL. The remineralization rate constants recalculated to the in situ temperature of 6℃ for labile and semi-labile DOC in pore water were 0.025 d-1 and 0.002 d-1, respectively, whereas, in bottom water, 0.026 d-1 and 0.004 d-1. The half-life times for DOCL were comparable for both bottom water and pore water and amounted to 26.2 d and 27.6 d, respectively. For DOCSL, the halflife time was shorter for bottom water (165.5 d) than for pore water (322.9 d). [ABSTRACT FROM AUTHOR]