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By using crawfish shells as the precursor and hydrothermal synthesis, Bovine serum albumin doped carbon dots (BSA@CDs) were prepared without excessive chemical reagents. The relationship between the fluorescence properties of different BSA@CDs and BSA amount was investigated by variouscharacterization techniques. When the amount of BSA added was 30 %, the prepared BSA@CDs' quantum yield (QY) reached 25.01 %, which was the highest. Inner Filter Effect (IFE) suggested that Cr (VI) can selectively quench the fluorescence of BSA@CDs. Cr (VI) can be reduced to Cr (III) by Hydroquinone (HQ), thus recovering the fluorescence. Accordingly, using BSA@CDs as a probe, a "turn-on" fluorescence sensor applied in HQ determination was constructed. The linear range was 10-200 µmol/L and limit of detection (LOD) was 0.18 µmol/L. Further, it has been employed to the determination of HQ in both crawfish tail meat and aquaculture water with good performance.

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The utilization of diagnostic-integrated molecules can enable targeted delivery and controlled release to significantly enhance therapeutic effectiveness and minimize toxic effects. Herein, we developed a novel class of glutathione (GSH)-activated bifunctional molecules that respond to elevated levels of GSH in tumor microenvironment. These bifunctional molecules retained the pharmacodynamic effects of parent molecules and mitigated cytotoxicity. Meanwhile, controlled release was monitored using fluorescent signals, enabling detection of drug distribution and accumulation in situ and in real time. Moreover, the correlation between GSH levels and fluorescence intensity offers the possibility of monitoring the effectiveness of responsive drugs. In conclusion, bifunctional molecules, as novel diagnostic-integrated molecules with both fluorescence imaging and therapeutic effects, exhibited potential applications in cancer therapy and imaging.

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This study introduces a novel fluorescence 'turn-on' chemosensor, FHDA, based on a trans-Ferulic acid Schiff-base derivative. FHDA stands out as a highly selective and sensitive tool for the fluorescent detection of Al3+ with the fluorescence 'turn-on' effect. FHDA exhibits a strong CHEF effect and ICT upon complexation with Al3+ in a 1:2 binding stoichiometry. The significant Stokes shift (Δλ = 108 nm, λex = 422 nm, λem = 530 nm), large binding constant (Ka = 4.2 × 104 M-1), ∼9.5-fold increase in the quantum yield (FHDA, Φ = 0.020; FHDA-Al3+ complex, Φ = 0.189), and a LOD of 134 nM, makes FHDA an excellent chemosensor for detecting Al3+ in solution; tests in live cells and environmental samples also showed excellent responses. FHDA offers substantial improvements over existing methods with its ease of use, limited expense, high specificity, and the ability to provide real-time, in-situ monitoring of Al3+ ions. The utility of FHDA is highlighted through applications in monitoring Al3+ ions in e.g. lung cancer cells (A549) and environmental water samples. We believe that applications of FHDA can potentially lead to a novel diagnostic and therapeutic strategy against diseases linked to aluminum dysregulation.

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The debate between the revisionist and received views of the relationship between Thomas Kuhn and logical empiricism has until now focused on the relationship between Kuhn and Rudolf Carnap. Here, I consider the relationship between Kuhn and two other members of the Vienna Circle's left-wing; Otto Neurath and Philipp Frank. It is argued that the attribution of the historical turn in philosophy of science to Kuhn obscures the historical awareness displayed in important works by members of the Vienna Circle, and thereby distorts its legacy. Both Frank and Neurath recognised the role for history in theorizing about science, and drawing upon these insights lead them to considerations of scientific theory-choice, rational disagreement, and the role of extra-scientific values in science, that anticipate those later made famous by Kuhn. It is also argued that the Left-Vienna Circle's programme for Unified Science, the replacement of traditional philosophy with a bipartite metatheory of science, provides a clearer and potentially more radical role for the history of science within the philosophy of science than Kuhn's. To reach this conclusion, it is demonstrated that some members of the Vienna Circle maintained a far less robust distinction between contexts of discovery and justification than has typically been attributed to them.

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Introduction: Athletes' capability to perform activities with body rotation could be weakened by fatigue accumulation. Making pivot turning in unanticipated scenarios after fatigue may greatly challenge athletes' ability to adapt rational motion strategies, elevating the risk of anterior cruciate ligament (ACL) injury. This study aimed to investigate the effects of fatigue and anticipation on biomechanical risk factors of ACL injury during 180° pivot turns in female soccer players. Methods: Twenty-one female soccer players were selected as participants. The participants performed anticipated turning maneuver before the fatigue intervention. The participants sprinted along the runway, decelerated and planted their foot on the force plate, and then executed a 180° pivot turn. For unanticipated tests, the pivot turn was mixed with side/cross-cuts, which were indicated to the participant using a custom-designed light system. The tests were repeated by the participant after receiving a fatigue intervention. Lower-limb joint angles and moments were characterized. Peak ground reaction forces (GRFs) and GRF loading rates were determined. Two-way repeated measures analysis of variance was applied to examine the effects of fatigue and anticipation on the variables of interest. Results: Compared to the anticipated conditions, the approach speed was significantly lower in the unanticipated tests (P < 0.0001). Lower-limb kinematics showed varied angular patterns across conditions: greater hip joint variations in flexion, abduction, and internal rotation during unanticipated turns; consistent knee joint flexion and ankle plantarflexion with dorsiflexion observed mid-turn. Significant interactions (P = 0.023 to P = 0.035) between fatigue and anticipation influenced hip joint angles. Anticipation effects were notable at initial contact and peak ground reaction force, increasing hip, knee, and ankle joint angles (P < 0.0001 to P = 0.012). Participants showed consistent ground reaction force (GRF) patterns during pivot turns across fatigue and anticipation conditions, with the first peak occurring approximately 10% into the turn period. Significant interaction effects (P = 0.016) between fatigue and anticipation were observed for knee flex/extension moments at the first peak vertical GRF. Anticipation significantly increased first peak vertical (P < 0.0001), anteroposterior (P < 0.0001), and mediolateral (P < 0.0001) GRFs. Fatigue increased first peak vertical (P = 0.022), anteroposterior (P = 0.018), and mediolateral (P = 0.019) GRFs. Post-fatigue, participants exhibited reduced first peak GRFs and loading rates compared to pre-fatigue conditions, with higher rates observed in unanticipated turns (vertical GRF: P = 0.030; anteroposterior GRF: P < 0.0001). Conclusion: Female soccer players' lower-limb Biomechanical characterization could be greatly affected by the change of anticipatory scenarios. With the associated increase of GRF, the risk of their ACL injury might be elevated. Fatigue affected female soccer players' abilities on movement performances, but the interaction of these two factors could potentially weaken their knee's functions during pivot turns. Cognitive training on unanticipated tasks may be important for rehabilitation training after ACL injury.

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The Escherichia coli cAMP receptor protein (CRP) relies on the F-helix, the recognition helix of the helix-turn-helix motif, for DNA binding. The importance of the CRP F-helix in DNA binding is well-established, yet there is little information on the roles of its non-base-contacting residues. Here, we show that a CRP F-helix position occupied by a non-base-contacting residue Val183 bears an unexpected importance in DNA binding. Codon randomization and successive in vivo screening selected six amino acids (alanine, cysteine, glycine, serine, threonine, and valine) at CRP position 183 to be compatible with DNA binding. These amino acids are quite different in their amino acid properties (polar, non-polar, hydrophobicity), but one commonality is that they are all relatively small. Larger amino acid substitutions such as histidine, methionine, and tyrosine were made site-directedly and showed to have no detectable DNA binding, further supporting the requirement of small amino acids at CRP position 183. Bioinformatics analysis revealed that small amino acids (92.15% valine and 7.75% alanine) exclusively occupy the position analogous to CRP Val183 in 1,007 core CRP homologs, consistent with our mutant data. However, in extended CRP homologs comprising 3700 proteins, larger amino acids could also occupy the position analogous to CRP Val183 albeit with low occurrence. Another bioinformatics analysis suggested that large amino acids could be tolerated by compensatory small-sized amino acids at their neighboring positions. A full understanding of the unexpected requirement of small amino acids at CRP position 183 for DNA binding entails the verification of the hypothesized compensatory change(s) in CRP.

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Human NAD(P)H-quinone oxidoreductase1 (HNQO1) is a two-electron reductase antioxidant enzyme whose expression is driven by the NRF2 transcription factor highly active in the prooxidant milieu found in human malignancies. The resulting abundance of NQO1 expression (up to 200-fold) in cancers and a barely detectable expression in body tissues makes it a selective marker of neoplasms. NQO1 can catalyze the repeated futile redox cycling of certain natural and synthetic quinones to their hydroxyquinones, consuming NADPH and generating rapid bursts of cytotoxic reactive oxygen species (ROS) and H2O2. A greater level of this quinone bioactivation due to elevated NQO1 content has been recognized as a tumor-specific therapeutic strategy, which, however, has not been clinically exploited. We review here the natural and new quinones activated by NQO1, the catalytic inhibitors, and the ensuing cell death mechanisms. Further, the cancer-selective expression of NQO1 has opened excellent opportunities for distinguishing cancer cells/tissues from their normal counterparts. Given this diagnostic, prognostic, and therapeutic importance, we and others have engineered a large number of specific NQO1 turn-on small molecule probes that remain latent but release intense fluorescence groups at near-infrared and other wavelengths, following enzymatic cleavage in cancer cells and tumor masses. This sensitive visualization/quantitation and powerful imaging technology based on NQO1 expression offers promise for guided cancer surgery, and the reagents suggest a theranostic potential for NQO1-targeted chemotherapy.

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Background: The Timed 360° turn test (T-360° TT) was developed to assess balance and turning ability. Although validity and reliability have been performed in different diseases, validity and reliability have not been performed in individuals with ankle sprain (AS). Purpose: The purpose of this study was to investigate the validity and reliability of the T-360° TT in individuals with AS. Methods: The study included 54 individuals with AS. Participants were initially evaluated with T-360° TT, Timed Up and Go (TUG) test and Biodex Balance System (BBS). To assess test-retest reliability, the T-360° TT was performed again 5 days after the first measurement by the same assessor. Results: At the end of the study, strong positive correlations were found between T-360° TT with TUG test and BBS (p < 0.05). In addition, T-360° TT had excellent test-retest reliability (Intraclass correlation coefficient = 0.87). Conclusion: The T-360° TT is a valid and reliable tool for the evaluation of balance and turning ability in individuals with AS. We also think that it can be used practically in clinical settings because it is a test that can be easily and quickly performed.

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In addition to the conventional chemotherapeutic drugs, potent inhibitors of key enzymes that are differentially overexpressed in cancer cells and associated with its progression are often considered as the drugs of choice for treating cancer. Aldose reductase (AR), which is primarily associated with complications of diabetes, is known to be closely related to the development of cancer and drug resistance. Epalrestat (EPA), an FDA-approved drug, is a potent inhibitor of AR and exhibits anticancer activity. However, its poor pharmacokinetic properties limit its bioavailability and therapeutic benefits. We report herein the first examples of esterase-responsive turn-on fluorogenic prodrugs for the sustained release of EPA to cancer cells with a turn-on fluorescence readout. Carboxylesterases are known to be overexpressed in several organ-specific cancer cells and help in selective uncaging of drug from the prodrugs. The prodrugs were synthesized using a multistep organic synthesis and successfully characterized. Absorption and emission spectroscopic studies indicated successful activation of the prodrugs in the presence of porcine liver esterase (PLE) under physiological condition. HPLC studies revealed a simultaneous release of both the drug and the fluorophore from the prodrugs over time with mechanistic insights. While the inhibitory potential of EPA released from the prodrugs toward the enzyme AR was validated in the aqueous medium, the anticancer activity of the prodrugs was studied in a representative cervical cancer cell line. Interestingly, our results revealed that the development of the prodrugs can significantly enhance the anticancer potential of EPA. Finally, the drug uncaging process from the prodrugs by the intracellular esterases was studied in the cellular medium by measuring the turn-on fluorescence using fluorescence microscopy. Therefore, the present study highlights the rational development of the fluorogenic prodrugs of EPA, which will help enhance its anticancer potential with better therapeutic potential.

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For over 40 years, the human consumption of dog meat has been a controversial issue in South Korea. While some defend it as cultural tradition, others criticize the act as unethical treatment of dogs. This study examined the public perceptions and opinions driving the legislative efforts to ban dog meat through an online survey of 1000 participants assessing their experiences with dog meat, awareness of related issues, and support for institutional bans. The results revealed a widespread negative view of the dog meat industry, largely due to concerns over animal cruelty, and a negligible demand for consuming dog meat, which points to the expected demise of the industry. However, there was notable resistance to the ban from the view that it restricts personal freedom, with anthropocentric attitudes hindering progress in animal welfare. Regression analysis identified gender, political affiliations, animal experiences, and perceptions of dog meat as key predictors of support for the ban. As beloved pets, dogs influence human ethics, and the growing awareness of animal ethics has led to a 'political turn' enabling policymakers to enact bans. This study highlights how animal welfare benefits from political actions backed by societal consensus.

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BACKGROUND: Generation and regulation (control) of linear and angular momentum is a challenge during turning while walking which may be exacerbated by age-related changes. In healthy older adults, little is known about how momentum is controlled during turns, especially within each phase of gait. Each phase of gait affords unique mechanical contexts to control momenta and regulate balance. In healthy young adults, we found that the transverse-plane linear and angular momenta generation strategies observed within specific phases of gait during straight-line gait were also used during turns. Therefore, in this study, we investigated whether healthy older adults shared similar momentum control strategies specific to each gait phase during straight-line gait and turns. METHODS: Nine healthy older adults completed straight-line gait and 90° leftward walking turns. We compared the change in transverse-plane whole-body linear and angular momentum across gait phases (left and right single and double support). We also compared the average leftward force and transverse-plane moment across gait phases. RESULTS: We found that leftward linear momentum was generated most during right single support in straight-line gait and leftward turns. However, in contrast to straight-line gait, during leftward turns, average leftward force was applied across gait phases, with left single support generating significantly less leftward average force than other gait phases. Leftward angular momentum generation and average moment were greatest during left double support in both tasks. We observed some within-participant results that diverged from the group statistical findings, illustrating that although they are common, these momenta control strategies are not necessary. CONCLUSIONS: Older adults generated transverse-plane linear and angular momentum during consistent phases of gait during straight-line gait and 90° turns, potentially indicating a shared control strategy. Understanding momentum control within each phase of gait can help design more specific targets in gait and balance training interventions.

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Beta turns, in which the protein backbone abruptly changes direction over four amino acid residues, are the most common type of protein secondary structure after alpha helices and beta sheets and play key structural and functional roles. Previous work has produced classification systems for turn geometry at multiple levels of precision, but these operate in backbone dihedral-angle (Ramachandran) space, and the absence of a local Euclidean-space coordinate system and structural alignment for turns, or of any systematic Euclidean-space characterization of turn backbone shape, presents challenges for the visualization, comparison and analysis of the wide range of turn conformations and the design of turns and the structures that incorporate them. This work derives a turn-local coordinate system that implicitly aligns turns, together with a set of geometric descriptors that characterize the bulk BB shapes of turns and describe modes of structural variation not explicitly captured by existing systems. These modes are shown to be meaningful by the demonstration of clear relationships between descriptor values and the electrostatic energy of the beta-turn H-bond, the overrepresentations of key side-chain motifs, and the structural contexts of turns. Geometric turn descriptors complement Ramachandran-space classifications, and they can be used to select turn structures for compatibility with particular side-chain interactions or contexts. Potential applications include protein design and other tasks in which an enhanced Euclidean-space characterization of turns may improve understanding or performance. The web-based tools ExploreTurns, MapTurns, and ProfileTurn, available at www.betaturn.com, incorporate turn-local coordinates and turn descriptors and demonstrate their utility.

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A Pro-Gly-typed cyclopeptide, cyclosenegalin A, was firstly isolated from Annona senegalensis and Annona scleroderma. In this study, we reported synthesis the cycloheptapeptide with a combination of solid- and solution-phase synthetic methods. This study also compared the effectiveness of ß-turn inducer type I and II in cyclosenegalin A to facilitate the cyclization process. The synthesis of cyclosenegalin A were prepared using two different sequences of linear peptides for cyclization. First sequence employed ß-turn type I inducer (Ser-Ala-Val-Thr) as turning point and second sequence employed ß-turn type II inducer (Thr-Pro-Gly-Leu). The successful cyclization was obtained using the linear sequence of NH2-Ala-Val-Thr-Pro-Gly-Leu-Ser-OH with ß-turn type II. The final product was obtained in 8.2 % yield with PyBOP/DIEA act as coupling agent. The synthetic cyclosenegalin A were characterized with HR-ToFMS, 1H NMR, 13C NMR, HSQC, HMBC, TOCSY, and ROESY. The synthetic product was also evaluated for its antimicrobial activity.

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This study compared laboratory risk and turn-around time (TAT) between sequence-based typing (SBT) and next-generation sequencing (NGS) for human leukocyte antigen (HLA) typing. For risk assessment, we utilized the risk priority number (RPN) score based on failure mode and effect analysis (FMEA) and a risk acceptability matrix (RAM) according to the Clinical Laboratory Standards Institute (CLSI) guidelines (EP23-A). Total TAT was documented for the analytical phase, and hands-on time was defined as manual processes conducted by medical technicians. NGS showed a significantly higher total RPN score than SBT (1169 vs. 465). NGS indicated a higher mean RPN score, indicating elevated severity and detectability scores in comparison to SBT (RPN 23 vs. 12, p = 0.001; severity 5 vs. 3, p = 0.005; detectability 5 vs. 4, p < 0.001, respectively). NGS required a greater number of steps than SBT (44 vs. 25 steps), all of which were acceptable for the RAM. NGS showed a longer total TAT, total hands-on time, and hands-on time per step than SBT (26:47:20 vs. 12:32:06, 03:59:35 vs. 00:47:39, 00:05:13 vs. 00:01:54 hh:mm:ss, respectively). Transitioning from SBT to NGS for HLA typing involves increased risk and an extended TAT. This study underscored the importance of evaluating these factors to optimize laboratory efficiency in HLA typing.

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RT-PCR is the gold standard for diagnosis of COVID-19. All RT-PCR kits are based on RNA extraction from the clinical sample. There was a sudden increase in demand of these kits, both RNA extraction and COVID-19 RT-PCR kits during the pandemic. This sudden spurt in global demand created a situation of shortage of consumables, especially the RNA extraction kits. Hence, this study was carried out to evaluate and compare COVID-19 RT-PCR without RNA extraction step using buffer R3. Sensitivity, specificity and accuracy of RT-PCR kit without RNA extraction were 89.16 %, 100% and 89.6% respectively. This approach saved more than 50 % time compared to the RT-PCR kit with RNA extraction approach allowing enhanced daily sample processing capability. RT-PCR kit without RNA extraction help in managing a greater number of samples, reduces cost and turnaround time.

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Head injury is a potentially lethal and frequently occurring condition in the emergency department (ED). Reliable and fast diagnosis is important both for patients and flow in the ED. Circulating S100B is used to rule out the need for head computer tomography in low-risk patients with mild head injury. The flow of these patients through the ED would benefit from shorter turn-around time. Standard serum clotting tubes require 30-60 min clotting time, followed by an analysis time of 45 min. Here, we evaluated the performance of two alternative blood collection tubes; a rapid serum tube (RST) with a recommend clotting time of 5 min and a hirudin tube (HIR) for instant anticoagulation. S100B measurement was performed on paired blood samples from 221 subjects using a Roche Cobas 602 analyser. The performances of the alternative tubes were evaluated by method comparison to the standard serum clotting tube, repeatability and agreement of results obtained from alternative tubes compared with the standard clotting tube. Both alternative tubes had a minor positive bias (RST = 0.011 µg/L, HIR = 0.008 µg/L). The repeatability was 2% for RST and 10% for HIR, while being 4% for the standard clotting tube. In the agreement analysis, the positive and negative predictive values for RST were 62% and 100% while being 73% and 99% for HIR respectively. Our study suggests that RST is a feasible alternative to reduce laboratory turn-around time in S100b analysis.

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A number of quinoidal molecules with symmetric end-capping groups, particularly dicyanomethylene units, have been synthesized for organic optoelectronic materials. In comparison, dissymmetric quinoidal molecules, characterized by end-capping with different groups, are less explored. In this paper, we present the unexpected formation of new formal quinoidal molecules, which are end-capped with both dicyanomethylene and triphenylphosphonium moieties. The structures of these dissymmetric quinoidal molecules were firmly verified by single crystal structural analyses. On the basis of the control experiments and DFT calculations, we proposed the reaction mechanism for the formation of these dissymmetric quinoidal molecules. The respective zwitterionic forms should make contributions to the ground state structures of these quinoidal molecules based on the analysis of their bond lengths and aromaticity and Mayer Bond Orbital (MBO) calculation. This agrees well with the fact that negative solvatochromism was observed for these quinoidal molecules. Although these new quinoidal molecules are non-emissive both in solutions and crystalline states, they become emissive with quantum yields up to 51.4% after elevating the solvent viscosity or dispersing them in a PMMA matrix. Interestingly, their emissions can also be switched on upon binding with certain proteins, in particular with human serum albumin.

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Mercury (Hg), a notorious heavy metal with detrimental impacts on human health and the environment, necessitates the development of precise measurement methods. This study introduces an expeditious and straightforward photochemical approach to synthesize thioglycolic acid (TGA)-stabilized CdTe/CdS/ZnS core/multi-shell quantum dots (QDs). The synthesized CdTe/CdS/ZnS QDs were comprehensively characterized using fluorescence spectroscopy, transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDS), Field Emission Scanning Electron Microscopy (FESEM), and X-Ray diffraction (XRD). XRD and EDS results confirmed the successful formation of CdTe/CdS/ZnS structure. Also, FESEM and TEM results showed that CdTe/CdS/ZnS QDs were spherical. Results showed that synthesized Exhibiting vibrant green fluorescence and notable quenching in the presence of Hg2+ ions, these QDs emerge as promising candidates for fabricating a fluorescent sensor. The proposed sensor demonstrates notable sensitivity to Hg2+, featuring a detection limit of 16.32 nM and a linear range from 20 nM to 70 nM. The sensor's selectivity was confirmed by analyzing various anions and cations. Moreover, when tested with tap water, river water, and agricultural samples, the sensor exhibited reliable performance, validated by Inductively Coupled Plasma (ICP) analysis. Additionally, CdTe/CdS/ZnS QDs immobilized on micro pads proved effective for on-site water sample analysis, presenting a versatile solution for environmental monitoring.

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Individuals with schizophrenia generally show difficulties in interpersonal communication. Linguistic analyses shed new light on speech atypicalities in schizophrenia. However, very little is known about conversational interaction management by these individuals. Moreover, the relationship between linguistic features, psychopathology, and patients' subjectivity has received limited attention to date. We used a novel methodology to explore dyadic conversations involving 58 participants (29 individuals with schizophrenia and 29 control persons) and medical doctors. High-quality stereo recordings were obtained and used to quantify turn-taking patterns. We investigated psychopathological dimensions and subjective experiences using the Positive and Negative Syndrome Scale for Schizophrenia (PANSS), the Examination of Anomalous Self Experience scale (EASE), the Autism Rating Scale (ARS) and the Abnormal Bodily Phenomena questionnaire (ABPq). Different turn-taking patterns of both patients and interviewers characterised conversations involving individuals with schizophrenia. We observed higher levels of overlap and mutual silence in dialogues with the patients compared to dialogues with control persons. Mutual silence was associated with negative symptom severity; no dialogical feature was correlated with anomalous subjective experiences. Our findings suggest that individuals with schizophrenia display peculiar turn-taking behaviour, thereby enhancing our understanding of interactional coordination in schizophrenia.

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The equilibrium state of hydrogen sulfide (H2S), a gaseous signaling molecule produced by lysosomal metabolites, in vivo is crucial for cellular function. Abnormal fluctuations in H2S concentration can interfere with the normal function of lysosomes, which has been closely linked to the pathogenesis of a variety of diseases. In view of this, a novel fluorescent probe Lyso-DPP based on 1,3,5-triarylpyrazolines was developed for the precise detection of H2S in lysosomes by using the hydrophilic morpholine moiety as a lysosomal targeting unit, and 2,4-dinitroanisole as a fluorescence-quenching and H2S-responsive unit. The probe cleverly combines the advantages of simple synthesis, sensitive blue fluorescence turn-on with a limit of detection, LOD, of 97.3 nM, good stability, and fast response time (10 min), which makes Lyso-DPP successful in portable monitoring of meat freshness in the form of test strips. Moreover, the excellent biocompatibility and precise targeting capability of the probe Lyso-DPP make it perform well in the monitoring of H2S in lysosomes, living cells, and zebrafish. This work not only provides new technical tools for food quality control but also paves up new ideas for early diagnosis and treatment of H2S-related diseases.

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