Conditioning Protocol Research Articles

Evaluation of Subclinical Cancer Therapy-Related Cardiac Dysfunction in Patients Undergoing Hematopoietic Stem Cell Transplantation: An Echocardiography Study

Background: Hematopoietic stem cell transplantation (HSCT) is a potentially curative procedure that is used in various hematological malignancies. However, among an increasing number of HSCT, the amount of cancer therapy-related cardiac dysfunction (CTRCD) is increasing as well. This study aimed to determine the prevalence of subclinical CTRCD in HSCT patients 12 months after HSCT and to assess the impact of clinical factors on the development of CTRCD. Material and Methods: We included 55 patients who underwent autologous or allogeneic HSCT. The patients were assessed using an echocardiography method before and 12 months after a HSCT procedure. Results: Our study revealed that during the 12-month follow-up period, asymptomatic CTRCD was observed in 15 patients (27.3%), 6 experienced moderate CTRCD, and 9 experienced mild CTRCD. Patients with previous use of anthracyclines tended to have CTRCD more often: nine patients (60%) in the CTRCD group and nine patients (22.5%) in non-CTRCD group. Patients who received the BEAM regimen for conditioning also experienced CTRCD more often: five patients (33.3%) in CTRCD group vs. two patients (5%) in the non-CTRCD group. Conclusions: Our study showed that asymptomatic CTRCD was found in 27.3% of the patients 12 months after HSCT. The BEAM chemotherapy conditioning protocol following prior anthracycline use were identified as factors contributing to the development of CTRCD.

A Conditional Privacy-Preserving Mutual Authentication Protocol With Fine-Grained Forward and Backward Security in IoV

A Conditional Privacy-Preserving Mutual Authentication Protocol With Fine-Grained Forward and Backward Security in IoV

Clinical and Electrocardiographic Predictors of Cardiac Resynchronization Therapy Response That Correlate with the 6 min Walking Test.

Background: Cardiac resynchronization therapy is an essential treatment for heart failure patients. Candidates typically have cardiomyopathy accompanied by delayed electrical activation in the left ventricular lateral wall, causing uncoordinated contractions and worsening heart failure. Heart failure severity can be assessed with functional tests: the cardiopulmonary test, which is a maximal exercise test, remains the gold standard, but the 6 min walk test has emerged as an easier, faster, and more comfortable alternative to be used by clinicians to adjust treatment protocols for cardiovascular and pulmonary conditions. Methods: This is a prospective observational study that included 69 patients from a single healthcare facility, and the purpose was to determine if the 6 min walk test results could be associated with changes in various electrocardiographic, clinical, functional, and demographic parameters. All the parameters and the 6 min walk distance were recorded at four key time moments: before the procedure and after 6, 9, and 12 months. The electrocardiographic parameters were obtained from the patients' electrocardiograms recorded in the four key moments and included variables such as QRS area, duration, percentage of biventricular pacing, and many others, while the functional variables included the monitored intraprocedural systolic blood pressure and the end-systolic left ventricular volume. We also aimed to check if clinical conditions such as diabetes and chronic kidney disease and demographic variables such as age or sex have any impact. Results and Conclusions: All this research was performed in order to identify which parameters hold a predictive value and can serve as future criteria for better patient selection and for defining a proper resynchronization outcome. The study shows that parameters such as diabetes and QRS duration have an impact over the 6 min walk distance. Also, newer variables such as the QRS area and the R/S ratio may represent a direction worth studying in order to predict the outcomes of cardiac resynchronization therapy.

Exploring the Impact of In Situ-Formed Solid-Electrolyte Interphase on the Cycling Performance of Aluminum Metal Anodes.

Unwanted processes in metal anode batteries, e.g., non-uniform metal electrodeposition, electrolyte decomposition, and/or short-circuiting, are not fully captured by the electrolyte bulk solvation structure but rather defined by the electrode-electrolyte interface and its changes induced by cycling conditions. Specifically, for aluminum-ion batteries (AIBs), the role of the solid-electrolyte interphase (SEI) on the Al0 electrodeposition mechanism and associated changes during resting or cycling remain unclear. Here, we investigated the current-dependent changes at the electrified aluminum anode/ionic liquid electrolyte interface to reveal the conditions of the SEI formation leading to irreversible cycling in the AIBs. We identified that the mechanism of anode failure depends on the nature of the counter electrode, where the areal capacity and cycling current for Al0 electrodeposition dictates the number of successful cycles. Notwithstanding the differences behind unstable aluminum anode cycling in symmetrical cells and AIBs, the uniform removal of electrochemically inactive SEI components, e.g., oxide-rich or solvent-derived organic-rich interphases, leads to more efficient cycling behavior. These understandings raise the importance of using specific conditioning protocols for efficient cycling of the aluminum anode in conjugation with different cathode materials.

Identification of transformation products from fluorinated lithium-ion battery additives TPFPB and TPFPP: forever chemicals of tomorrow?

Fluorinated organic compounds (FOCs) represent a class of synthetic chemicals distinguished by their resilient carbon-fluorine bonds, which demonstrate an ability to withstand environmental degradation over an extended period. The integration of FOCs into cutting-edge applications, including lithium-ion batteries (LiBs), presents considerable potential for environmental harm that has not yet been sufficiently addressed. This study focuses on the environmental fate of two fluorinated aromatics, tris(pentafluorophenyl)borane (TPFPB) and tris(pentafluorophenyl)phosphine (TPFPP), given their important role in improving the performance of LiBs. To achieve this, laboratory simulation methods including total oxidizable precursor assay, electrochemistry (EC), Fenton reaction, UV-C irradiation, and hydrolysis were employed. Liquid chromatography and gas chromatography coupled with high-resolution mass spectrometry were used for identification of transformation products (TPs) and prediction of their molecular formulae. Despite the structural similarity between TPFPB and TPFPP, distinct differences in electrochemical behavior and degradation pathways were observed. TPFPB readily underwent hydroxylation and hydrolysis, resulting in a wide range of 49 TPs. A total of 28 TPs were newly identified, including oligomers and highly toxic dioxins. In contrast, TPFPP degraded exclusively under harsh conditions, requiring the development of innovative conditioning protocols for EC. In total, the simulation experiments yielded nine structurally different compounds, including seven previously undescribed, partially defluorinated TPs. This study highlights the potential risks associated with the use of FOCs in LiBs and provides insight into the complex environmental behavior of FOCs.

Multimodal assessment of the role of intolerance of uncertainty in fear acquisition and extinction

Personality traits linked to internalizing disorders influence the way we develop fears, but also how we regain a sense of safety. In the present study, we investigated the effect of intolerance of uncertainty (IU) on defensive responses using a differential fear conditioning protocol with an extinction phase. The conditioned stimulus was associated with an aversive sound (90 dB) in 75 % of the presentations during acquisition. A final sample of 176 participants completed the experiment. We measured self-reports of associative (expectancy of the unconditioned stimulus in acquisition) and evaluative learning (arousal and valence), and both physiological (skin conductance response) and electrocortical responses (steady-state visually evoked potentials, ssVEPs; late positive potentials, LPP) to the conditioned stimuli. Our results show that IU's impact is limited, with no effect in both acquisition and extinction. These findings emphasize the necessity of large samples in research on inter-individual differences and contribute to our understanding of how IU may or may not be involved in fear and safety learning processes considering multiple aspects of fear responding.

Analysis of fast calcium dynamics of honey bee olfactory coding

Odour processing exhibits multiple parallels between vertebrate and invertebrate olfactory systems. Insects, in particular, have emerged as relevant models for olfactory studies because of the tractability of their olfactory circuits. Here, we used fast calcium imaging to track the activity of projection neurons in the honey bee antennal lobe (AL) during olfactory stimulation at high temporal resolution. We observed a heterogeneity of response profiles and an abundance of inhibitory activities, resulting in various response latencies and stimulus-specific post-odour neural signatures. Recorded calcium signals were fed to a mushroom body (MB) model constructed implementing the fundamental features of connectivity between olfactory projection neurons, Kenyon cells (KC), and MB output neurons (MBON). The model accounts for the increase of odorant discrimination in the MB compared to the AL and reveals the recruitment of two distinct KC populations that represent odorants and their aftersmell as two separate but temporally coherent neural objects. Finally, we showed that the learning-induced modulation of KC-to-MBON synapses can explain both the variations in associative learning scores across different conditioning protocols used in bees and the bees' response latency. Thus, it provides a simple explanation of how the time contingency between the stimulus and the reward can be encoded without the need for time tracking. This study broadens our understanding of olfactory coding and learning in honey bees. It demonstrates that a model based on simple MB connectivity rules and fed with real physiological data can explain fundamental aspects of odour processing and associative learning.

Platelets as Regenerative Cells, Hundreds of Opportunities in Sight

<strong>Platelets were the last to be discovered of</strong> the three formed blood elements. Several scientists have been interested in them, recognizing them as cells other than leukocytes and erythrocytes. Its job in hemostasis and thrombosis is currently known. Platelets contain growth factors and secrete cytokines, which have roles in inflammation and tissue regeneration processes. This is why platelet concentrates have been widely used in regenerative medicine, an emerging field that involves biomolecules, cells, and scaffolds. Due to their chemotactic, angiogenic, immunomodulatory, and cell differentiation-generating properties, platelets have been used to promote the regeneration of tissues such as bones, tendons, ligaments, and wounds. The efficacy and safety of platelet-rich plasma (PRP) have been demonstrated in various medical scenarios. However, it is essential to implement standardized protocols for each population's conditions and conduct clinical studies based on the available evidence. It must be considered that, as a biological product, the number of platelets used, the number of growth factors, and the way of application will lead to results that will depend on each patient.

Understanding the Impact of Conditioning Protocols for Water Electrolyzers on Durability

The global push towards climate neutrality has intensified research efforts on green hydrogen production technologies. Among them, proton exchange water electrolysis (PEMWE) stands out as a highly efficient technology for producing green hydrogen using renewable energies. Nevertheless, a significant reduction in the cost of PEMWE while maintaining its durability over its operational periods is imperative. The DOE’s Hydrogen Earthshot target aims to achieve $1 per 1kg of H2 production within the decade. Reducing the amount of expensive iridium catalysts also reduces the durability of PEMWE. Hence, understanding the degradation mechanisms of low Ir loading PEMWE holds utmost importance. One key step in the evaluation of PEMWE is the break-in or conditioning step. Ensuring optimal conditioning is crucial to guarantee the peak performance of membrane electrode assembly (MEA) before assessing the durability of the catalyst thereby ensuring representative and reproducible results.This research aims to comprehensively evaluate various conditioning protocols for PEMWE and their impact on durability. Utilizing a combination of electrochemical, electron, and X-ray characterization techniques. Accelerated stress tests (AST) will be conducted after each conditioning protocol to evaluate the degradation rate of the catalyst. Ex-situ characterization of the MEA post-conditioning as well as post-AST offers a comprehensive understanding of how the catalyst’s initial state, such as the iridium oxidation state, following the conditioning step directly influences the durability. The goal is to develop an optimal conditioning protocol for PEMWE, that considers the performance and durability challenges associated with low catalyst loading. Acknowledgment This research is supported by the U.S. Department of Energy (DOE) Hydrogen and Fuel Cell Technologies Office through the Hydrogen from Next-generation Electrolyzers of Water (H2NEW) consortium with support from Dave Peterson and McKenzie Hubert.

Fear generalization modulated by shock intensity and protein synthesis inhibitor.

Maladaptive fear responses, including sensitized threat reactions and overgeneralization, contribute to anxiety disorders such as generalized anxiety disorder and post-traumatic stress disorder. Although stress intensity influences the generation and extent of these maladaptive fears, the underlying mechanisms remain unclear. The present study examined whether varying footshock stress intensity and inhibition of protein synthesis have differential effect on fear sensitization and generalization in mice. Mice were subjected to a classic fear conditioning protocol involving five different levels of footshock intensities. Prior to fear acquisition, the protein synthesis inhibitor cycloheximide (CHX) was administered intraperitoneally. Fear sensitization to white noise and fear generalization to tones with frequencies differing from the conditioned tone were assessed at either 2 or 4 days after fear acquisition. The results showed that, although varying shock intensities (except the lowest) led to a similar pattern of increased freezing during auditory cues in fear acquisition, the extent of both fear sensitization and generalization increased with the intensity of the footshock in the following days. As shock intensities increased, there was a proportional rise in sensitized fear to white noise and generalized freezing to tones with frequencies progressively closer to the conditioned stimulus. Mildest shocks did not induce discriminative conditioned fear memory, whereas the most intense shocks led to pronounced fear generalization. Administration of CHX before fear acquisition did not affect sensitized fear but reduced generalization of freezing to tones dissimilar from the conditioned stimulus in the group exposed to the most intense shock. Our results suggest that maladaptive fear responses elicited by varying stress intensities exhibit distinct characteristics. The effect of CHX to prevent overgeneralization without affecting discriminative fear memory points to potential therapeutic approaches for fear-related disorders, suggesting the possibility of mitigating overgeneralization while preserving necessary fear discrimination.

Sex-dependent effects of amyloidosis on functional network hub topology is associated with downregulated neuronal gene signatures in the APPswe/PSEN1dE9 double transgenic mouse.

Extracellular beta-amyloid (Aβ) is thought to cause impairments in brain-wide functional connectivity, although mechanisms linking Aβ to broader functional network processing remain elusive. In the present study, we evaluated the effects of Aβ on fear memory and functional connectome measures in male and female mice. Middle-aged (9-11mo) double transgenic APP-PS1 mice and age and sex-matched controls were tested on a fear conditioning protocol and then imaged at 11.1 Tesla. Brains were harvested and processed for analysis of Aβ plaques and Iba1 immunolabeling in neocortical areas, hippocampus, and basolateral amygdala. Additional RNA sequencing data from separate age, strain, and sex-matched mice were analyzed for differentially expressed genes (DEGs) and weighted gene co-expression networks. In both male and female mice, we observed increased functional connectivity in a dorsal striatal/amygdala network as a result of Aβ. Increased functional connectivity within this network was matched by increases in APP gene expression, Aβ and Iba1 immunolabeling, and an upregulated cluster of DEGs involved in the immune response. Conversely, the network measure representing node hubness, eigenvector centrality, was increased in prefrontal cortical brain regions, but only in female APP-PS1 mice. This female-specific effect of amyloid was associated with down-regulation of a cluster of DEGs involved in cortical and striatal GABA transmission, anxiogenic responses, and motor activity, in female APP-PS1 mice, but not males. Our results contribute to a growing literature linking between Aβ, immune activation, and functional network connectivity. Furthermore, our results reveal outcomes of Aβ on gene expression patterns in female mice that may contribute to amyloidosis-induced dysregulation of non-cognitive circuitry.

ELEVATING ATHLETIC PERFORMANCE: THE BODY TECHNIQUE PROTOCOL'S EFFECT ON VITALITY AND RESPIRATORY WELLNESS

Engaging in regular exercise and sports can inadvertently lead to habitual movement patterns, restricting specific body parts' range of motion and diminishing joint elasticity. This decline in muscle flexibility impacts energy levels during physical activities, thus affecting overall vitality. Subjective Vitality State (SVS) captures the essence of feeling alive, reflecting an individual's energy reserves. The study involved 36 healthy young athletes aged 22.48±3.54 years, categorized into control and treatment groups. The treatment group followed a novel Body Technique exercise protocol (BT), while the control group adhered to a familiar conditioning protocol. Both protocols included five exercises targeting various muscle groups. The study focused on subjective vitality, perceived intensity load, and chest girth differences during breathing. Analysis of subjective vitality change within groups showed no significant difference for the control group before and after treatment (p=0.44), while the treatment group exhibited significantly higher subjective vitality after BT exercises (p=0.00). Significant differences were found in the "exhale-inhale" variable for the treatment group before and after treatment (p=0.00), but not for the control group (p=0.15). BT protocol demonstrated superior effects on SVS and chest girth, highlighting the significance of innovative, unconventional exercise in sports. By addressing concerns such as fatigue and injury, BT protocol may offer the prospect of better results in forthcoming training sessions. The study encourages a paradigm shift in exercise approaches, emphasizing conscious intervention to address novel movement patterns and enhance well-being in athletes.

Contextual fear conditioning in zebrafish: Influence of different shock frequencies, context, and pharmacological modulation on behavior

Contextual fear conditioning is a protocol used to assess associative learning across species, including fish. Here, our goal was to expand the analysis of behavioral parameters that may reflect aversive behaviors in a contextual fear conditioning protocol using adult zebrafish (Danio rerio) and to verify how such parameters can be modulated. First, we analyzed the influence of an aversive stimulus (3 mild electric shocks for 5 s each at frequencies of 10, 100 or 1000 Hz) on fish behavior, and their ability to elicit fear responses in the absence of shock during a test session. To confirm whether the aversive responses are context-dependent, behaviors were also measured in a different experimental environment in a test session. Furthermore, we investigated the effects of dizocilpine (MK-801, 2 mg/kg, i.p.) on fear-related responses. Zebrafish showed significant changes in baseline activity immediately after shock exposure in the training session, in which 100 Hz induced robust contextual fear responses during the test session. Importantly, when introduced to a different environment, animals exposed to the aversive stimulus did not show any differences in locomotion and immobility-related parameters. MK-801 administered after the training session reduced fear responses during the test, indicating that glutamate NMDA-receptors play a key role in the consolidation of contextual fear-related memory in zebrafish. In conclusion, by further exploring fear-related behaviors in a contextual fear conditioning task, we show the effects of different shock frequencies and confirm the importance of context on aversive responses for associative learning in zebrafish. Additionally, our data support the use of zebrafish in contextual fear conditioning tasks, as well as for advancing pharmacological studies related to associative learning in translational neurobehavioral research.

Influence of ageing and moisture conditions on the ravelling resistance of the micro-surfacing mix

ABSTRACT In this study, the durability of the micro-surfacing mix was assessed by investigating the synergistic effect of ageing and moisture conditioning on ravelling. A total of 30 combinations of conditioning protocols were considered. Ageing time and temperature were varied between 85–105°C and 24–120 h, whereas moisture conditioning time and temperature were varied between 25–60°C and 24–192 h. The results showed that after the mix was fully cured, increased stiffness led to a substantial decrease in ravelling. Synergistic assessment of ageing and moisture showed that ageing temperature had the highest, whereas moisture conditioning time had the least influence on ravelling. It was noticed that once the specimens were fully cured and subjected to further ageing, the effect of moisture conditioning temperature on ravelling became minimal. Overall, laboratory investigations pointed out that micro-surfacing should be applied during favourable environmental conditions, facilitating faster moisture loss and strength gain to improve durability.

Non-Hebbian plasticity transforms transient experiences into lasting memories.

The dominant models of learning and memory, such as Hebbian plasticity, propose that experiences are transformed into memories through input-specific synaptic plasticity at the time of learning. However, synaptic plasticity is neither strictly input-specific nor restricted to the time of its induction. The impact of such forms of non-Hebbian plasticity on memory has been difficult to test, and hence poorly understood. Here, we demonstrate that synaptic manipulations can deviate from the Hebbian model of learning, yet produce a lasting memory. First, we established a weak associative conditioning protocol in mice, where optogenetic stimulation of sensory thalamic input to the amygdala was paired with a footshock, but no detectable memory was formed. However, when the same input was potentiated minutes before or after, or even 24 hr later, the associative experience was converted into a lasting memory. Importantly, potentiating an independent input to the amygdala minutes but not 24 hr after the pairing produced a lasting memory. Thus, our findings suggest that the process of transformation of a transient experience into a memory is neither restricted to the time of the experience nor to the synapses triggered by it; instead, it can be influenced by past and future events.

Contextual pain conditioning protocol design in Humans – a combination of behavioral and physiological measurements using immersive Virtual Reality

Contextual pain conditioning protocol design in Humans – a combination of behavioral and physiological measurements using immersive Virtual Reality

Neuropsychological and Anatomical-Functional Effects of Transcranial Magnetic Stimulation in Post-Stroke Patients with Cognitive Impairment and Aphasia: A Systematic Review.

Transcranial magnetic stimulation (TMS) has been found to be promising in the neurorehabilitation of post-stroke patients. Aphasia and cognitive impairment (CI) are prevalent post-stroke; however, there is still a lack of consensus about the characteristics of interventions based on TMS and its neuropsychological and anatomical-functional benefits. Therefore, studies that contribute to creating TMS protocols for these neurological conditions are necessary.To analyze the evidence of the neuropsychological and anatomical-functional TMS effects in post-stroke patients with CI and aphasia and determine the characteristics of the most used TMS in research practice.The present study followed the PRISMA guidelines and included articles from PubMed, Scopus, Web of Science, ScienceDirect, and EMBASE databases, published between January 2010 and March 2023.In the 15 articles reviewed, it was found that attention, memory, executive function, language comprehension, naming, and verbal fluency (semantic and phonological) are the neuropsychological domains that improved post-TMS. Moreover, TMS in aphasia and post-stroke CI contribute to greater frontal activation (in the inferior frontal gyrus, pars triangularis, and opercularis). Temporoparietal effects were also found. The observed effects occur when TMS is implemented in repetitive modality, at a frequency of 1Hz, in sessions of 30min, and that last more than 2 weeks in duration.The use of TMS contributes to the neurorehabilitation process in post-stroke patients with CI and aphasia. However, it is still necessary to standardize future intervention protocols based on accurate TMS characteristics.

Behavioral and genetic correlates of heterogeneity in learning performance in individual honeybees, Apis mellifera.

Learning an olfactory discrimination task leads to heterogeneous results in honeybees with some bees performing very well and others at low rates. Here we investigated this behavioral heterogeneity and asked whether it was associated with particular gene expression patterns in the bee's brain. Bees were individually conditioned using a sequential conditioning protocol involving several phases of olfactory learning and retention tests. A cumulative score was used to differentiate the tested bees into high and low performers. The rate of CS+ odor learning was found to correlate most strongly with a cumulative performance score extracted from all learning and retention tests. Microarray analysis of gene expression in the mushroom body area of the brains of these bees identified a number of differentially expressed genes between high and low performers. These genes are associated with diverse biological functions, such as neurotransmission, memory formation, cargo trafficking and development.

Conditioning of Motor Evoked Responses After Anterior Cruciate Ligament Reconstruction: Effects of Stimulus Intensity.

Operant conditioning of motor evoked torque (MEPTORQUE) can directly target the corticospinal pathway in patients with anterior cruciate ligament (ACL) reconstruction. However, it remains unclear whether operant conditioning can elicit short-term improvements in corticospinal excitability and whether these improvements are influenced by stimulus intensity. Quadriceps MEPTORQUE responses can be upconditioned in a single session and will elicit short-term adaptations in corticospinal excitability, with higher stimulus intensities eliciting greater effects. Randomized controlled laboratory study. Level 2. Thirty-six participants were assessed during a single session of an operant conditioning protocol. Participants were randomized into 1 of 3 groups for stimulus intensity used during operant conditioning based on the participant's active motor threshold (AMT: 100%, 120%, and 140%). Quadriceps MEPTORQUE amplitude was evaluated during a block of control transcranial magnetic stimulation trials (CTRL) to establish baseline corticospinal excitability, and 3 blocks of conditioning trials (COND) during which participants trained to upcondition their MEPTORQUE. MEPTORQUE recruitment curves were collected to evaluate the effect of operant conditioning on acute corticospinal adaptations. Participants with ACL reconstruction could upcondition their MEPTORQUE in a single session (P < 0.01; CTRL, 17.27 ± 1.28; COND, 21.35 ± 1.28 [mean ± standard error [SE] in N·m]), but this ability was not influenced by the stimulus intensity used during training (P = 0.84). Furthermore, significant improvements in corticospinal excitability were observed (P = 0.05; PRE, 687.91 ± 50.15; POST, 761.08 ± 50.15 [mean ± SE in N·m %AMT]), but stimulus intensity did not influence corticospinal adaptations (P = 0.67). Operant conditioning can elicit short-term neural adaptations in ACL-reconstructed patients. Future operant conditioning paradigms may effectively use any of the 3 stimulus intensities studied herein. Operant conditioning may be a feasible approach to improve corticospinal excitability after ACL reconstruction.

Advancing Field Aging Simulation Methods for Cantabro Abrasion Loss Testing

Over the past several years, the asphalt industry has progressively increased efforts to evaluate mixtures after laboratory conditioning for the purpose of simulating field aging, and has also increased reliance on mixture testing. AASHTO T 401-22 describes Cantabro Mass Loss testing and has an appendix of field aging simulation protocols that can be used to estimate field aging by making use of compacted specimens. T 401-22 is one of the few methods to contain both a field aging simulation protocol and a mixture property index test. This paper uses roughly 1,000 tested specimens from forty-nine mixes (mostly dense-graded asphalt with minor amounts of stone-matrix asphalt) evaluated for up to four years of field aging in a hot and non-freeze climate alongside twenty-five different laboratory conditioning protocols. This paper’s objective is to provide information to improve T 401-22 by way of recommendations to update the appendix of conditioning protocols with more comprehensive assessments of their effectiveness in simulating field aging in a hot, non-freeze climate. The overarching recommendation of this work culminates with four protocols. Two of these protocols use only oven conditioning, whereas the remaining two protocols use a combination of stages exposing specimens to oxidation in ovens, moisture effects in hot water baths, and freeze–thaw cycling. The time required to administer the recommended protocols varies from approximately 6 to 28 days and they simulate between 1 and 5 years of field aging depending on the protocol selected.