Ex vivo analysis of cortical microarchitecture of the distal clavicle: implications for surgical management of fractures.

BACKGROUND: Cortical thickness and porosity are two main determinants of cortical bone strength. Thus, mapping variations in these parameters across the full width of the distal end of the clavicle may be helpful for better understanding the basis of distal clavicle fractures and for selecting optimal surgical treatment. METHODS: Distal ends of 11 clavicles (6 men, 5 women; age: 81.9 ± 15.1 years) were scanned by micro-computed tomography at 10-µm resolution. We first analyzed cortical thickness and porosity of each 500-µm-wide area across the superior surface of distal clavicle at the level of conoid tubercle in an antero-posterior direction. This level was chosen for detailed evaluation because previous studies have demonstrated its superior microarchitecture relative to the rest of the distal clavicle. Subsequently, we divided the full width of distal clavicle to three subregions (anterior, middle, and posterior) and analyzed cortical porosity, pore diameter, pore separation, and cortical thickness. RESULTS: We found the largest number of low-thickness and high-porosity areas in the anterior subregion. Cortical porosity, pore diameter, pore separation, and cortical thickness varied significantly among the three subregions (p < 0.001 p = 0.016, p = 0.001, p < 0.001, respectively). Cortex of the anterior subregion was more porous than that of the middle subregion (p < 0.001) and more porous and thinner than that of the posterior subregion (p < 0.001, p = 0.030, respectively). Interaction of site and sex revealed higher porosity of the anterior subregion in women (p < 0.001). The anterior subregion had larger pores than the middle subregion (p = 0.019), whereas the middle subregion had greater pore separation compared with the anterior (p = 0.002) and posterior subregions (p = 0.006). In general, compared with men, women had thinner (p < 0.001) and more porous cortex (p = 0.03) with larger cortical pores (p < 0.001). CONCLUSIONS: Due to high cortical porosity and low thickness, the anterior conoid subregion exhibits poor bone microarchitecture, particularly in women, which may be considered in clinical practice. LEVELS OF EVIDENCE: Level IV.

Search for new biologically active compounds: in vitro studies of antitumor and antimicrobial activity of dirhodium(II,II) paddlewheel complexes.

Four neutral Rh1-Rh4 complexes of the general formula [Rh2(CH3COO)4L2], where L is an N-alkylimidazole ligand, were synthesized and characterized using various spectroscopic techniques, and in the case of Rh4 the crystal structure was confirmed. Investigation of the interactions of these complexes with HSA by fluorescence spectroscopy revealed that the binding constants Kb are moderately strong (∼104 M-1), and site-marker competition experiments showed that the complexes bind to Heme site III (subdomain IB). Competitive binding studies for CT DNA using EB and HOE showed that the complexes bind to the minor groove, which was also confirmed by viscosity experiments. Molecular docking confirmed the experimental data for HSA and CT DNA. Antimicrobial tests showed that the Rh2-Rh4 complexes exerted a strong inhibitory effect on G+ bacteria B. cereus and G- bacteria V. parahaemolyticus as well as on the yeast C. tropicalis, which showed a higher sensitivity compared to fluconazole. The cytotoxic activity of Rh1-Rh4 complexes tested on three cancer cell lines (HeLa, HCT116 and MDA-MB-231) and on healthy MRC-5 cells showed that all investigated complexes elicited more efficient cytotoxicity on all tested tumor cells than on control cells. Investigation of the mechanism of action revealed that the Rh1-Rh4 complexes inhibit cell proliferation via different mechanisms of action, namely apoptosis (increase in expression of the pro-apoptotic Bax protein and caspase-3 protein in HeLa and HCT116 cells; changes in mitochondrial potential and mitochondrial damage; release of cytochrome c from the mitochondria; cell cycle arrest in G2/M phase in both HeLa and HCT116 cells together with a decrease in the expression of cyclin A and cyclin B) and autophagy (reduction in the expression of the protein p62 in HeLa and HCT116 cells).

The role of phenotypic plasticity and corticosterone in coping with pond drying conditions in yellow-bellied toad (Bombina variegata, Linnaeus 1758) tadpoles.

Amphibian larvae inhabiting temporary ponds often exhibit the capacity to accelerate development and undergo metamorphosis in challenging conditions like desiccation. However, not all species exhibit this ability, the yellow-bellied toad (Bombina variegata) is one such example. The underlying mechanisms behind the inability to accelerate development under desiccation remain largely unexplored. The hypothalamic-pituitary-interrenal (HPI) axis and corticosterone (CORT), which act synergistically with thyroid hormone, are thought to facilitate metamorphosis in response to desiccation stress. In this study, we aimed to investigate whether modification in the HPI axis, particularly CORT levels, contributes to the absence of adaptive plasticity in B. variegata under desiccation stress. The study design included four treatments: high water level, high water level with exogenous CORT, low water level, and low water level with metyrapone (a CORT synthesis inhibitor). The main objective was to evaluate the effects of these treatments on whole-body corticosterone levels, life history, morphological traits, and oxidative stress parameters during the prometamorphic and metamorphic climax developmental stages. While low water level had no effect on total corticosterone levels, larval period, body condition index, and metamorphic body shape, it negatively affected metamorph size, mass, and growth rate. Our findings suggest that constant exposure to desiccation stress over generations may have led to modifications in the HPI axis activity in B. variegata, resulting in adaptation to changes in water level, evident through the absence of stress response. Consequently, CORT may not be a relevant stress indicator in desiccation conditions for this species.

Potential application of green extracts rich in phenolics for innovative functional foods: natural deep eutectic solvents as media for isolation of biocompounds from berries.

The health-promoting effects of berries have attracted attention due to the possible application of their extracts as functional ingredients in food products. Natural deep eutectic solvents (NADESs) are a new generation of environmentally friendly solvents for the extraction of natural products, and they are green alternatives to organic solvents, and they can improve the solubility, stability, and bioavailability of isolated biocompounds. In this study, an efficient eco-friendly method was used for the extraction of phenolic compounds from different berries: chokeberries, blueberries, and black goji berries with a range of eutectic solvents consisting of hydrogen bond acceptors (HBAs) such as choline chloride, L-proline, L-glycine, and L-lysine and hydrogen bond donors (HBDs) such as malic, citric, tartaric, lactic and succinic acids, glucose and glycerol. The obtained results indicated the ability of NADESs towards selective extraction of phenolics; the eutectic system choline chloride : malic acid showed selective extraction of anthocyanins, while choline chloride : glycerol and choline chloride : urea showed selectivity towards flavonoids and phenolic acids. The methodology for screening of the NADES extraction performance, which included chromatographic profiling via high-performance thin layer chromatography combined with chemometrics and spectrophotometric essays, allowed effective assessment of optimal eutectic solvents for isolation of different groups of phenolics. Great antioxidant and antimicrobial activities of extracts, along with the green nature of eutectic solvents, enable NADES berry extracts to be used as "green-labelled" functional foods or ingredients.

Applying machine learning in the investigation of the link between the high-velocity streams of charged solar particles and precipitation-induced floods.

This study explores a possible link between solar activity and floods caused by precipitation. For this purpose, discrete blocks of data for 89 separate flood events in Europe in the period 2009-2018 were used. Solar activity parameters with a time lag of 0-11 days were used as input data of the model, while precipitation data in the 12 days preceding the flood were used as output data. The level of randomness of the input and output time series was determined by correlation analysis, while the potential causal relationship was established by applying machine learning classification predictive modeling. A total of 25 distinct machine-learning algorithms and four model ensembles were applied. It was shown that in 81% of cases, the designed model could explain the occurrence or absence of precipitation-induced floods 9 days in advance. Differential proton flux in the 0.068-0.115 MeV and integral proton flux > 2.5 MeV were found to be the most important factors for forecasting precipitation-induced floods. The study confirmed that machine learning is a valuable technique for establishing nonlinear relationships between solar activity parameters and the onset of floods induced by precipitation.

Does the anesthetic tricaine methanesulfonate (MS-222) distort oxidative status parameters in tadpoles?

The effect of anesthesia/euthanasia with ethyl 3-aminobenzoate methanesulfonate (MS-222) on the oxidative status of Hyla arborea tadpoles was examined to determine whether the use of the anesthetic can confound the experimental results of the oxidative stress-based investigation. The experiment was conducted on two groups of tadpoles reared at different temperatures to produce differences in antioxidant capacity between the groups. After development at different temperatures (20 °C and 25 °C), the animals were exposed to different concentrations of MS-222 (0, 0.1, 1, and 5 g/L) for 15 min. The higher temperature decreased catalase activity, glutathione and protein carbonyl levels and increased glutathione reductase activity. The glutathione level and glutathione/thiol-related parameters were significantly changed after MS-222 exposure. However, individuals from the different temperature groups responded differently to the tested anesthetic, pointing to the possible influence of the initial levels of antioxidant capacity. The analysis of the interaction between the factors (temperature and MS-222) confirmed that the anesthetic can confound the results regarding the effects of temperature on the oxidative status parameters. The concentration of 0.1 g/L MS-222 had the lowest influence on the alterations in oxidative status and the results of the effect of temperature. A brief review of the current literature on the use of MS-222 in tadpoles made clear the absence of precise information on anesthetic concentration and exposure time. Similar studies should be repeated and extended to other amphibian species and other factors of interest to provide better guidance on tadpole anesthesia/euthanasia for future experiments that consider oxidative status parameters.

Ligand coupling mechanism of the human serotonin transporter differentiates substrates from inhibitors.

The presynaptic serotonin transporter (SERT) clears extracellular serotonin following vesicular release to ensure temporal and spatial regulation of serotonergic signalling and neurotransmitter homeostasis. Prescription drugs used to treat neurobehavioral disorders, including depression, anxiety, and obsessive-compulsive disorder, trap SERT by blocking the transport cycle. In contrast, illicit drugs of abuse like amphetamines reverse SERT directionality, causing serotonin efflux. Both processes result in increased extracellular serotonin levels. By combining molecular dynamics simulations with biochemical experiments and using a homologous series of serotonin analogues, we uncovered the coupling mechanism between the substrate and the transporter, which triggers the uptake of serotonin. Free energy analysis showed that only scaffold-bound substrates could initiate SERT occlusion through attractive long-range electrostatic interactions acting on the bundle domain. The associated spatial requirements define substrate and inhibitor properties, enabling additional possibilities for rational drug design approaches.

Bioassay-Guided Assessment of Antioxidative, Anti-Inflammatory and Antimicrobial Activities of Extracts from Medicinal Plants via High-Performance Thin-Layer Chromatography.

Natural products and their analogues have contributed significantly to treatment options, especially for anti-inflammatory and infectious diseases. Thus, the primary objective of this work was to compare the bioactivity profiles of selected medicinal plants that are historically used in folk medicine to treat inflammation and infections in the body. Chemical HPTLC fingerprinting was used to assess antioxidant, phenolic and flavonoid content, while bioassay-guided HPTLC was used to detect compounds with the highest antibacterial and anti-inflammatory activities. The results of this study showed that green tea leaf, walnut leaf, St. John's wort herb, wild thyme herb, European goldenrod herb, chamomile flower, and immortelle flower extracts were strong radical scavengers. Green tea and nettle extracts were the most active extracts against E. coli, while calendula flower extract showed significant potency against S. aureus. Furthermore, green tea, greater celandine, and fumitory extracts exhibited pronounced potential in suppressing COX-1 activity. The bioactive compounds from the green tea extract, as the most bioactive, were isolated by preparative thin-layer chromatography and characterized with their FTIR spectra. Although earlier studies have related green tea's anti-inflammatory properties to the presence of catechins, particularly epigallocatechin-3-gallate, the FTIR spectrum of the compound from the most intense bioactive zone showed the strongest anti-inflammatory activity can be attributed to amino acids and heterocyclic compounds. As expected, antibacterial activity in extracts was related to fatty acids and monoglycerides.

Novel synthetic procedures for C2 substituted imidazoquinolines as ligands for the α/ß-interface of the GABAA-receptor.

A series of substituted imidazoquinolines, a structurally related chemotype to pyrazoloquinolinones, a well-known class of GABAA ligands, was prepared via two synthetic procedures and the efficiency of these procedures were compared. One method relies on classical heterocyclic synthesis, the other one aims at late-stage decoration of a truncated scaffold via direct C-H functionalization. A pharmacological evaluation disclosed that one of the synthesized derivatives showed interesting activity on a α1ß3 containing receptor subtype. Supplementary Information: The online version contains supplementary material available at 10.1007/s00706-022-02988-8.

Novel pyrazolothienopyridinones as potential GABAA receptor modulators.

The synthesis of novel pyrazolothienopyridinone derivatives as potential GABAA receptor modulators was performed and is herein described. A crucial step of the synthesis involving handling unstable aminothiophenes was managed via two different synthetic strategies delivering a set of 8 target compounds. Supplementary Information: The online version contains supplementary material available at 10.1007/s00706-023-03063-6.

Singlet and Triplet Pathways Determine the Thermal Z/E Isomerization of an Arylazopyrazole-Based Photoswitch.

Understanding the thermal isomerization mechanism of azobenzene derivatives is essential to designing photoswitches with tunable half-lives. Herein, we employ quantum chemical calculations, nonadiabatic transition state theory, and photosensitized experiments to unravel the thermal Z/E isomerization of a heteroaromatic azoswitch, the phenylazo-1,3,5-trimethylpyrazole. In contrast to the parent azobenzene, we predict two pathways to be operative at room temperature. One is a conventional ground-state reaction occurring via inversion of the aryl group, and the other is a nonadiabatic process involving intersystem crossing to the lowest-lying triplet state and back to the ground state, accompanied by a torsional motion around the azo bond. Our results illustrate that the fastest reaction rate is not controlled by the mechanism involving the lowest activation energy, but the size of the spin-orbit couplings at the crossing between the singlet and the triplet potential energy surfaces is also determinant. It is therefore mandatory to consider all of the multiple reaction pathways in azoswitches in order to predict experimental half-lives.

Effectiveness of Eccentric Overload Training in Basketball Players: A Systematic Review.

Several studies have confirmed the efficacy of flywheel eccentric overload training in order to improve or increase muscle volume and strength, as well as several performance-related fitness attributes, but to date, there are no studies that have reviewed the effects of these training methods in basketball. Therefore, the present systematic review aimed to collect the updated information about the influence of flywheel training on performance of basketball players. A search in four electronic databases (PubMed, SCOPUS, Web of Science, and Google Scholar) was conducted up to November 20, 2022. Articles were selected as valid for review if: (i) they were an experimental trial published in English; (ii) participants were basketball players without illnesses or injuries, and (iii) a rotational inertial overload method was used as a performance-dependent variable. A total of 93 articles were found. After filtering procedures, only seven studies were considered in this systematic review. In the selected studies, 203 participants were included, 58 females and 145 males. Focusing on basketball related performance variables, all the studies included the countermovement jump (CMJ), while five included sprinting and a change of direction test (COD). The results showed an improvement in performance-related variables associated with basketball (sprint, CMJ, and COD) through the use of inertial methods. Improvements were reported in professional, semi-professional and amateur players as well as both male and female players. However, given the low number of studies, additional investigations on this topic are warranted before a "clear picture" can be drawn concerning the effects of flywheel eccentric overload training in basketball players.

Effects of Eccentric-Oriented Strength Training on Return to Sport Criteria in Late-Stage Anterior Cruciate Ligament (ACL)-Reconstructed Professional Team Sport Players.

Background and Objectives: An effective post-injury training program is essential to regain performance and fulfill criteria for return to sport for team sport athletes following anterior cruciate ligament (ACL) reconstruction. The aim of this study was to compare the effects of 6 weeks of eccentric-oriented strength training vs. traditional strength training during the late-stage ACL-rehab phase on leg strength and vertical and horizontal jumping performance in professional team sport athletes. Materials and Methods: Twenty-two subjects (14 males, 8 females, age 19.9 ± 4.4 years, mass 77.4 ± 15.6 kg, height 182.4 ± 11.7 cm) (mean ± SD) with a unilateral reconstructed ACL (BTB graft) were included in the study. All participants enrolled in the same rehabilitation protocol prior to the training study. Players were randomly assigned to an experimental (ECC: n = 11, age 21.8 ± 4.6 years, mass 82.7 ± 16.6 kg, height 185.4 ± 12.2 cm), and a control group (CON: n = 11, age 19.1 ± 2.1 years, mass 76.6 ± 16.5 kg, height 182.5 ± 10.2 cm). Both groups underwent an equivolumed rehabilitation program, with the only difference being in strength training, which consisted of flywheel training vs. traditional strength training for the experimental and control groups, respectively. Testing was organized before and after the 6-week training programs and included isometric semi-squat tests (ISOSI-injured and ISOSU-uninjured legs), vertical jump tests (CMJ), single-leg vertical jump tests (SLJI-injured and SLJU-uninjured legs), single-leg hop tests (SLHI-injured and SLHU-uninjured legs), and triple hop tests (TLHI-injured and TLHU-uninjured legs). In addition, limb symmetry indexes were calculated for the isometric semi-squat (ISOSLSI) test, the single-leg vertical jump (SLJLSI), and the hop (SLHLSI) tests, as well as the triple-leg hop (THLLSI) test. Results: Main effects of time across training were observed for all dependent variables (posttest > pretest, p < 0.05). Significant group-by-time interactions were found for ISOSU (p < 0.05, ES = 2.51, very large), ISOSI (p < 0.05, ES = 1.78, large), CMJ (p < 0.05, ES = 2.23, very large), SLJI (p < 0.05, ES = 1.48, large), SLHI (p < 0.05, ES = 1.83, large), and TLHI (p < 0.05, ES = 1.83, large). Conclusions: This study suggests that eccentric-oriented strength training in late-stage ACL recovery, undertaken twice or three times weekly for 6 weeks, results in better outcomes than traditional strength training in leg strength, vertical jump ability, and single and triple hop tests with injured legs in professional team sport athletes. It seems that flywheel strength training can be recommended in late-stage ACL recovery for professional team sport athletes in order to regain recommended performance outcome levels faster.

Changes of oxidative status in yellowfin seabream larvae (Acanthopagrus latus) during development.

Dynamic changes of tissues, organs and growth that occur in fish larvae during the transition to the juvenile stage are accompanied by differences in metabolic, locomotor and feeding activities that can reflect on fish's oxidative status. In this study, we examine how body growth, antioxidant system (superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and total antioxidant capacity (TAC) and oxidative damage (malondialdehyde-MDA) parameters change in larvae of yellowfin seabream larvae (Acanthopagrus latus) during early development (0, 7, 15, 22 and 30 day after hatching-DAH). Body growth (length and weight) starts to intensify from 15 DAH. We observed general increase in the antioxidant system (AOS) with the age of larvae from newly hatched and 7 DAH up to 15 and 22 DAH individuals. 15 and 22 DAH larvae had the greatest levels of TAC, SOD and GR activity, while 30 DAH larvae had higher CAT activity from 0, 7 and 15 DAH and MDA concentration in comparison to 15 DAH individuals. Several developmental events can be linked with observed results: lower AOS in 0 and 7 DAH individuals with low locomotor activity, growth, endo-exogenous feeding phase and cell differentiation; 15 and 22 DAH larvae are under pressure of fast growth, enhanced swimming and foraging capacity; while higher MDA production in 30 DAH larvae can be a result of shifts in muscle metabolism, changes in both quality and quantity of food and a significant increase in weight. The present study provides insight into the changes in redox status during the ontogeny of A. latus, fish species about which physiology is still little known but with a potential for use in marine culture. Ability to lower oxidative stress during critical developmental periods can enhance that potential.

Effects of cadmium chloride and biofertilizer (Bacilar) on biochemical parameters of freshwater fish, Alburnus mossulensis.

Fish in wild are often faced with various types of xenobiotics, that may display synergistic or antagonistic effects. In this study, we aim to examine how exposure to agrochemical compound (Bacilar) and cadmium (CdCl2) alone and in combination affect biochemical parameters (lactate dehydrogenase, aspartate aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, alanine aminotransferase; creatine phosphokinase (CKP), cholinesterase) and oxidative stress (total antioxidant capacity, catalase, malondialdehyde and protein carbonyl concentrations) of freshwater fish Alburnus mossulensis. Fish were exposed to two concentrations of Bacilar (0.3, and 0.6 mL L-1) and to 1 mg L-1 cadmium chloride alone and in combination for 21 days. Results showed that fish accumulate Cd in their body, with the highest rate in individuals exposed to Cd in combination with Bacilar. Both xenobiotics in fish liver induced the activation of liver enzymes suggesting hepatotoxic effects, with the greatest impact in co-exposed groups. A significant decrease in the hepatocyte's total antioxidant capacity indicates the collapse of the antioxidant defense in fish exposed to Cd and Bacilar. A decrease in the antioxidant biomarkers was followed by increased oxidative damage of lipids and proteins. We also reported altered function in the muscle of individuals exposed to Bacilar and Cd seen as decreased activities in CKP and butyrylcholinesterase. Overall, our results point to the toxicity of both Bacilar and Cd on fish but also to their synergistic effects on Cd bioaccumulation, oxidative stress, and liver and muscle damage. This study highlights the need for evaluating the use of agrochemicals and their possible additive effects on non-target organisms.

Evaluation of outdoor thermal comfort conditions: evidence from the Serbian major ski resort over the last 30 years.

The study aims to present reliable information about thermal conditions and their impacts on visitors to ski travel destinations. Mountain tourism areas are specific since high altitudes affect the ambient weather conditions which can affect different types of human activities. In this paper, the thermal comfort and its changes in Kopaonik Mountain, the most popular ski resort in Serbia over the last 30 years, have been evaluated. Information about thermal comfort is presented by using the Universal Thermal Climate Index (UTCI), physiologically equivalent temperature (PET), and modified physiologically equivalent temperature (mPET) in 3-h resolution for the period 1991-2020. The results indicate prevailing cold stress all year round. Days with moderate, strong, and very strong heat stress were not recorded. Strong and extreme cold stress prevailed during winter, while slight and moderate cold stress prevailed during summer. Transitional seasons were very cold, but autumn was more comfortable than spring. The occurrence of days with neutral and slightly warm/cool conditions is concentrated in the summer months. However, summer is not used enough for tourism because the choice of tourists to stay at Kopaonik is not primarily based on favorable bioclimatic conditions, but on resources for winter tourism. With global warming, the annual number of thermally favorable days has been increasing, while the number of days with extreme and strong cold stress is decreasing. Continuing this trend can significantly influence tourism in the future, and therefore, new strategies in ski resorts will be required to adapt to the changing climate.

Enlightening the Path to Protein Engineering: Chemoselective Turn-On Probes for High-Throughput Screening of Enzymatic Activity.

Many successful stories in enzyme engineering are based on the creation of randomized diversity in large mutant libraries, containing millions to billions of enzyme variants. Methods that enabled their evaluation with high throughput are dominated by spectroscopic techniques due to their high speed and sensitivity. A large proportion of studies relies on fluorogenic substrates that mimic the chemical properties of the target or coupled enzymatic assays with an optical read-out that assesses the desired catalytic efficiency indirectly. The most reliable hits, however, are achieved by screening for conversions of the starting material to the desired product. For this purpose, functional group assays offer a general approach to achieve a fast, optical read-out. They use the chemoselectivity, differences in electronic and steric properties of various functional groups, to reduce the number of false-positive results and the analytical noise stemming from enzymatic background activities. This review summarizes the developments and use of functional group probes for chemoselective derivatizations, with a clear focus on screening for enzymatic activity in protein engineering.

Higher temperature induces oxidative stress in hybrids but not in parental species: A case study of crested newts.

Ectotherms are particularly sensitive to global warming due to their limited capacity to thermoregulate, which can impact their performance and fitness. From a physiological standpoint, higher temperatures often enhance biological processes that can induce the production of reactive oxygen species and result in a state of cellular oxidative stress. Temperature alters interspecific interactions, including species hybridization. Hybridization under different thermal conditions could amplify parental (genetic) incompatibilities, thus affecting a hybrid's development and distribution. Understanding the impact of global warming on the physiology of hybrids and particularly their oxidative status could help in predicting future scenarios in ecosystems and in hybrids. In the present study, we investigated the effect of water temperature on the development, growth and oxidative stress of two crested newt species and their reciprocal hybrids. Larvae of Triturus macedonicus and T. ivanbureschi, and their T. macedonicus-mothered and T. ivanbureschi-mothered hybrids were exposed for 30 days to temperatures of 19°C and 24°C. Under the higher temperature, the hybrids experienced increases in both growth and developmental rates, while parental species exhibited accelerated growth (T. macedonicus) or development (T. ivanbureschi). Warm conditions also had different effects on the oxidative status of hybrid and parental species. Parental species had enhanced antioxidant responses (catalase, glutathione peroxidase, glutathione S-transferase and SH groups), which allowed them to alleviate temperature-induced stress (revealed by the absence of oxidative damage). However, warming induced an antioxidant response in the hybrids, including oxidative damage in the form of lipid peroxidation. These findings point to a greater disruption of redox regulation and metabolic machinery in hybrid newts, which can be interpreted as the cost of hybridization that is likely linked to parental incompatibilities expressed under a higher temperature. Our study aims to improve mechanistic understanding of the resilience and distribution of hybrid species that cope with climate-driven changes.

What coloration brings: Implications of background adaptation to oxidative stress in anurans.

BACKGROUND: Urban development results in habitat destruction, affecting populations of amphibians, the most fragile group of vertebrates. With changes in the environment, these animals become more exposed to light and predators. To enhance their chances of survival, they display plasticity of body coloration. Aside from adaptive benefits, animals exhibiting background matching meet the energetic costs and restrictions of changing body tones. To study the physiological consequences of Hyla arborea tadpole adaptation to background color, we followed oxidative stress parameters after rearing larvae on a constant background (black/white) and after changing the background color. RESULTS: Larvae cultivated for 20 days on constant substrate color exhibited differences in body coloration but without differences in lipid peroxidation (LPO) concentration between dark and pale individuals, suggesting that coloration investment during this period did not induce higher oxidative damage in darker tadpoles. Prolonged exposure of larvae (37 days) to a dark habitat increased antioxidative system defense and LPO concentrations, compared to animals reared permanently in the white surroundings. The positive correlation of oxidative damage with color intensity of individuals points to the physiological consequences of higher investment in the number of pigment cells necessary for dark pigmentation. In individuals faced with non-matching background and change in body coloration, defense system declined and LPO occurred relative to individuals cultivated in white habitat. CONCLUSION: Here, we have pointed to consequences related to background matching and stress that amphibians experienced during chromatic adaptations. Background color change causes a complex physiological response affecting the antioxidative defense parameters. This investigation elucidates the accompanying cost of amphibians' adjustment to an altered environment.

Cerebellar α6GABAA Receptors as a Therapeutic Target for Essential Tremor: Proof-of-Concept Study with Ethanol and Pyrazoloquinolinones.

Ethanol has been shown to suppress essential tremor (ET) in patients at low-to-moderate doses, but its mechanism(s) of action remain unknown. One of the ET hypotheses attributes the ET tremorgenesis to the over-activated firing of inferior olivary neurons, causing synchronic rhythmic firings of cerebellar Purkinje cells. Purkinje cells, however, also receive excitatory inputs from granule cells where the α6 subunit-containing GABAA receptors (α6GABAARs) are abundantly expressed. Since ethanol is a positive allosteric modulator (PAM) of α6GABAARs, such action may mediate its anti-tremor effect. Employing the harmaline-induced ET model in male ICR mice, we evaluated the possible anti-tremor effects of ethanol and α6GABAAR-selective pyrazoloquinolinone PAMs. The burrowing activity, an indicator of well-being in rodents, was measured concurrently. Ethanol significantly and dose-dependently attenuated action tremor at non-sedative doses (0.4-2.4 g/kg, i.p.). Propranolol and α6GABAAR-selective pyrazoloquinolinones also significantly suppressed tremor activity. Neither ethanol nor propranolol, but only pyrazoloquinolinones, restored burrowing activity in harmaline-treated mice. Importantly, intra-cerebellar micro-injection of furosemide (an α6GABAAR antagonist) had a trend of blocking the effect of pyrazoloquinolinone Compound 6 or ethanol on harmaline-induced tremor. In addition, the anti-tremor effects of Compound 6 and ethanol were synergistic. These results suggest that low doses of ethanol and α6GABAAR-selective PAMs can attenuate action tremor, at least partially by modulating cerebellar α6GABAARs. Thus, α6GABAARs are potential therapeutic targets for ET, and α6GABAAR-selective PAMs may be a potential mono- or add-on therapy.