Nonspecific Effects Research Articles

Modern Pain Science and Alexander Technique: How Might Alexander Technique Reduce Pain?

This article brings together research from the fields of pain science and Alexander Technique (AT) to investigate the mechanisms by which AT helps reduce pain. AT is a cognitive embodiment practice and a method for intentionally altering habitual postural behavior. Studies show that AT helps with various kinds of pain, although the mechanisms of pain reduction are currently not well understood. Advances in pain science may give insight into how this occurs. Modern interventions with efficacy for improving pain and function are consistent with active approaches within kinesiology. They also share similarities with AT and may have common mechanisms such as learning, mind–body engagement, normalization of sensorimotor function, improvement of psychological factors, and self-efficacy, as well as nonspecific treatment effects. AT likely has additional unique mechanisms, including normalization of muscle tone, neuronal excitability, and tissue loading, as well as alterations to body schema, attention redirection, and reduction in overall reactivity.

Most Carpal Tunnel Releases Address Moderate or Severe Median Neuropathy.

The role of surgery in normal or very mild median neuropathy can be questioned given that surgery in the absence of pathophysiology may offer only nonspecific effects that can be achieved without surgery, which raises ethical concerns. It's also important to avoid misdiagnosis: given that mild median neuropathy is prevalent and generally well-accommodated, notable symptoms from mild median neuropathy can signal unhelpful thoughts and feelings of distress that are a more pressing health priority. We identified 38 studies that categorized electrodiagnostic (EDX) measured median neuropathy severity among people being evaluated for carpal tunnel syndrome. We converted the different EDX grades used into one general grading classification. The Kruskal-Wallis test was used to compare the ratio of moderate/severe to mild/normal EDX findings between tests ordered by a surgeon or a nonoperative clinician. The median (interquartile range) ratio of moderate/severe to mild/normal EDX findings of median neuropathy at the carpal tunnel was 1.2 (0.91 to 1.8) among tests ordered by nonoperative clinicians and 3.3 (1.5 to 5.3) among tests ordered by surgeons (P = .0023). Only 4.1% of the patients who had EDX testing ordered by a surgeon had no measurable neuropathy. There were zero patients with normal EDX findings in 10 of the 17 (59%) studies in surgeon practices and in 10 of the 21 (48%) in the practices of nonoperative clinicians. The observation that in case series documenting EDX severities of median neuropathy, surgeons are mostly treating and operating on moderate to severe pathophysiology, emphasizes that while mild median neuropathy is highly prevalent it is uncommonly considered for surgery. Level II, Prognostic study.

Randomized Controlled Trials Studying Nonoperative Treatments of Osteoarthritis Often Use Misleading and Uninformative Control Groups: A Systematic Review.

Because there are no known treatments that alter the natural course of the pathophysiology of osteoarthritis, nonoperative treatment needs to be compared with known effective treatments that seek to mitigate symptoms or with similarly invasive inert (placebo) treatments to determine effectiveness. Comparing a treatment to an uninformative control group may inappropriately legitimize and support the use of potentially ineffective treatments. We therefore investigated the prevalence of inappropriate control groups in musculoskeletal research and asked whether these are associated with reporting a positive treatment effect. We systematically reviewed randomized trials of nonoperative treatments of osteoarthritis and asked: (1) What proportion of randomized trials use uninformative control groups (defined as a treatment less invasive than the tested treatment, or a treatment that might possibly not outperform placebo but is not acknowledged as such)? (2) Is the use of uninformative control groups independently associated with reporting a positive treatment effect (defined as p < 0.05 in favor of the intervention, or as making a recommendation favoring the intervention over the control treatment)? In a systematic review following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, we searched PubMed, Cochrane, and Embase up to September 2023 for randomized controlled trials published between 2020 to 2022 that compared one or more nonoperative treatments for the symptoms of osteoarthritis. We excluded studies that contained a surgical treatment group. We identified 103 trials that met eligibility criteria, with a total of 15,491 patients. The risk of bias was high in 60% (n = 62) of trials using the Cochrane Risk of Bias Tool, version 2. Although the high risk of bias in the included studies is concerning, it does not invalidate our design; instead, it highlights that some studies may use flawed methods to recommend treatments with unproven effectiveness beyond nonspecific effects because the kinds of bias observed would tend to increase the apparent benefit of the treatment(s) being evaluated. We used logistic regression to test the association of uninformative control groups with a positive treatment effect, accounting for potential confounders such as conflict of interest and study bias using the Cochrane Risk of Bias score. The use of uninformative control groups (treatments less invasive than the tested treatment, or treatments that might not outperform placebo but are not acknowledged as such) was found in 46% (47 of 103) of included studies. After accounting for potential confounding, there was no association between reporting positive treatment effects and the use of an uninformative control group. Studies with a low risk of bias had a lower likelihood of reporting a positive treatment effect (OR 0.2 [95% confidence interval 0.05 to 0.9]; p = 0.04, model pseudo R2 = 0.21). The finding that recent studies that mimic high-level evidence often use uninformative control groups that do not adequately account for nonspecific effects (perceived treatment benefits unrelated to a treatment's direct physiological effects) points to a high risk of legitimizing ineffective treatments. This raises the ethical imperative for patients, clinicians, journal peer reviewers, and journal editors to hold researchers to the standard of an adequate, informative control group. Awareness and risk of bias checklists might help patients and clinicians forgo new treatments based on seemingly high-level evidence that may carry only iatrogenic, financial, and psychological harm (false hope, in particular). Level I, therapeutic study.

Correlation Of BCG Vaccination And COVID-19 Disease Through Measurement Of Immunological Markers IFN-Γ And Anti-BCG Igg In Different Age Groups Running Title: Correlation Between BCG Vaccination And COVID-19 Disease

Introduction Coronavirus disease (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in China in December 2019 and then became a pandemic. BCG vaccination has conferred non-specific (targeted-off) protective effects against unrelated pathogens including respiratory viral infections that have been hypothesized to it could reduce the severity of COVID-19 disease. Objective: Assess whether BCG vaccination has a role in the prevention of COVID-19. Methods: A cross-sectional study including 111 healthy people was carried out from October 2021 to December 2022. Specific anti-BGC-IgG and IFN-γ levels were estimated in blood samples after stimulation with BCG vaccine for 2 days. All participants were examined for BCG scar status and COVID-19 confirmation. Results: There was no significant difference in anti-BCG IgG levels or IFN-γ levels (p&gt;0.05) in relation to COVID-19 infection. There was no statistically significant association between BCG scar diameter and COVID-19 infection (p&gt;0.05). Furthermore, there was no statistically significant association between the BCG scar diameter and COVID-19 severity (p&gt;0.05). There was no significant difference in anti-BCG IgG levels (p&gt;0.05) in relation to COVID-19 severity. Likewise, there was no significant difference in between IFN γ levels and COVID-19 severity (p&gt;0.05). Conclusion: There was no correlation between BCG vaccination and infection or severity of COVID-19.

Sodium Ion-Induced Structural Transition on the Surface of a DNA-Interacting Protein.

Protein surfaces have pivotal roles in interactions between proteins and other biological molecules. However, the structural dynamics of protein surfaces have rarely been explored and are poorly understood. Here, the surface of a single-stranded DNA (ssDNA) binding protein (SSB) with four DNA binding domains that bind ssDNA in binding site sizes of 35, 56, and 65 nucleotides per tetramer is investigated. Using oligonucleotides as probes to sense the charged surface, NaCl induces a two-state structural transition on the SSB surface even at moderate concentrations. Chelation of sodium ions with charged amino acids alters the network of hydrogen bonds and/or salt bridges on the surface. Such changes are associated with changes in the electrostatic potential landscape and interaction mode. These findings advance the understanding of the molecular mechanism underlying the enigmatic salt-induced transitions between different DNA binding site sizes of SSBs. This work demonstrates that monovalent salt is a key regulator of biomolecular interactions that not only play roles in non-specific electrostatic screening effects as usually assumed but also may configure the surface of proteins to contribute to the effective regulation of biomolecular recognition and other downstream events.

Comparison of in vitro screening methods for evaluating the effects of pharmaceutical excipients on membrane permeability

The effects of pharmaceutical excipients on intestinal drug absorption have been highlighted and careful excipient selection is required to develop biologically equivalent formulations. This study aimed to evaluate the effects of excipients on drug permeability and compare the characteristics of in vitro screening methods. Three in vitro models, the commercial precoated parallel artificial membrane permeability assay (PAMPA), PermeaPadTM, and Caco-2 monolayer, were used to evaluate the effects of 14 excipients on the permeability of several drugs with different biopharmaceutical classification system classes. Concentration-dependent effects were analyzed to distinguish non-specific effects. The permeability of low-permeability drugs was increased by excipients such as hydroxypropyl cellulose and povidone K30 in the precoated PAMPA model, whereas PermeaPadTM maintained membrane integrity at higher concentrations. Conversely, croscarmellose sodium and sodium lauryl sulfate (SLS) decreased the permeability of highly permeable drugs in both precoated PAMPA and PermeaPadTM assays in a concentration-dependent manner. In Caco-2 monolayer assays, most excipients showed minimal effects on drug permeability. However, SLS significantly reduces the permeability of highly permeable drugs at concentrations above the critical micelle concentration, thereby compromising the integrity of the cell monolayer. Our results suggested that most of excipients, except SLS, did not affect the membrane permeation of drugs at clinically used concentrations. The pre-coated PAMPA model demonstrated high sensitivity to excipient effects, making it suitable for conservative evaluation. The PermeaPadTM and Caco-2 models allowed assessment at higher excipient concentrations, with PermeaPadTM being particularly useful for excipients that cause toxicity in Caco-2 cells.

Peptide-Amphiphilic Nanoassemblies as a Responsive Senolytic Navigator for Targeted Removal of Senescent Cardiomyocytes to Ameliorate Heart Failure.

Heart failure (HF) represents the terminal stage of numerous cardiovascular disorders and lacks effective therapeutic strategies. The accumulation of senescent cardiomyocytes is a cardinal characteristic of HF, contributing to myocardial dysfunction and deteriorating the myocardial microenvironment through the development of senescence-associated secretory phenotypes (SASPs), ultimately culminating in pathological remodeling. Senolytics, a promising therapeutic strategy that selectively induces apoptosis in senescent cells, faces challenges due to nonspecific effects, raising concerns for clinical implementation. In this study, we developed peptide-amphiphilic nanoassemblies as responsive drug navigators for targeted delivery. The modular nanoassemblies comprise a hydrophilic domain containing a CD9-binding peptide, a hydrophobic domain incorporating a reactive oxygen species (ROS)-responsive motif, and an alkyl tail for encapsulation of the senolytic ABT263. The CD9-targeted and ROS-responsive nanoassemblies (AP@ABT263) specifically recognized senescent cardiomyocytes and modulated the release of ABT263 in the presence of elevated intracellular ROS levels. AP@ABT263 treatment significantly enhanced the targeted delivery of ABT263 to senescent cells in both in vitro and in vivo while showing minimal toxicity to normal cardiomyocytes and other tissues. Our findings provide compelling evidence that AP@ABT263 efficiently eradicated senescent cardiomyocytes, enhanced cardiac function, and attenuated the deleterious effects of SASP, thereby preventing adverse cardiac remodeling. In summary, AP@ABT263 represents a highly promising approach for responsive and controlled drug release in senescent cardiomyocytes, providing valuable insights into the development of intelligent pharmaceutical interventions for the management of HF.

Pyrolytic conversion of glucose into hydroxymethylfurfural and furfural: Benchmark quantum-chemical calculations.

Quantum chemical methods have been intensively applied to study the pyrolytic conversion of glucose into hydroxymethylfurfural (HMF) and furfural (FF). Herein, we collect the most relevant mechanistic proposals from the recent literature and organize them into a single reaction network. All the transition structures (TSs) and intermediates are characterized using highly accurate ab initio methods and the possible reaction pathways are assessed in terms of the Gibbs energies of the TSs and intermediates with respect to β-glucopyranose, selecting a 2D ideal-gas standard state at 773 K to represent the pyrolysis conditions. Several pathways can lead to the formation of both HMF and FF passing through rate-determining TSs that have ΔG‡ values of ~49-50 kcal/mol. Both water-assisted mechanisms and nonspecific environmental effects have a minor impact on the Gibbs energy profiles. We find that the HMF → FF + CH2O fragmentation has a small ΔrxnG value and an accessible ΔG‡ barrier. Our computational results, which are in consonance with the kinetic parameters derived from lumped models, the results of isotopic labeling experiments and the reported HMF/FF molecular ratios, could be useful for modeling studies including on nonequilibrium kinetic effects that may render more information about product yields and the relevance of the various pathways.

Identification of Major Interfering Substances for Heavy Metal Determination in Northern Sea Areas of Sri Lanka

This study investigated the physicochemical characteristics of seawater from five locations along the northern Sri Lankan coast, encompassing areas with varying degrees of anthropogenic activity. The analysis revealed significant variations in several parameters, potentially influenced by human influences. The results reveal significant variations in several parameters. For instance, turbidity levels varied from 1.88 NTU in Thalaiyadi to 13.3 NTU in Pannai, with Pannai, Myliddy, and Kakkaithivu exceeding the recommended limit of 5 NTU. Total suspended solids (TSS) ranged from 3 mg/L in Thalaiyadi to 83 mg/L in Pannai, surpassing the recommended limit of 30 mg/L. Electrical conductivity (EC) ranged from 45,700 µS/cm in Nainathivu to 49,500 µS/cm in Pannai, exceeding the typical seawater range. Nitrate levels ranged from 10.4 mg/L in Thalaiyadi to 19.1 mg/L in Myliddy. Major cations such as calcium (361 mg/L to 417 mg/L), magnesium (1,222 mg/L to 1,327 mg/L), and sodium (10,140 mg/L to 10,530 mg/L) also showed significant differences across locations. These findings highlight the need for continuous monitoring and effective management strategies to mitigate the negative impacts of anthropogenic activities on these coastal ecosystems. Furthermore, the complex seawater matrix presents challenges for heavy metal determination using Graphite Furnace Atomic Absorption Spectroscopy (GFAAS) due to spectral interferences from major cations, non-specific matrix effects, and interferences from organic matter, turbidity, and suspended solids. Careful consideration of these factors through background correction techniques, matrix modifiers, sample pre-treatment, and optimization of analysis parameters is crucial for accurate heavy metal determination in these environments. This study contributes to a better understanding of the environmental conditions and emphasizes the need for further research on heavy metal contamination and the development of robust analytical methods tailored to address the challenges posed by the complex seawater matrix in northern Sri Lanka.

Longitudinal development of resting-state functional connectivity in adolescents with and without internalizing disorders

Longitudinal studies using resting-state functional magnetic resonance imaging (rs-fMRI) focused on adolescent internalizing psychopathology are scarce and have mostly investigated standardized treatment effects on functional connectivity (FC) of the full amygdala. The role of amygdala subregions and large resting-state networks had yet to be elucidated, and treatment is in practice often personalized. Here, longitudinal FC development of amygdala subregions and whole-brain networks are investigated in a clinically representative sample.Treatment-naïve adolescents with clinical depression and comorbid anxiety who started care-as-usual (n = 23; INT) and healthy controls (n = 24; HC) participated in rs-fMRI scans and questionnaires at baseline (before treatment) and after three months. Changes between and within groups over time in FC of the laterobasal amygdala (LBA), centromedial amygdala (CMA) and whole-brain networks derived from independent component analysis (ICA) were investigated.Groups differed significantly in FC development of the right LBA to the postcentral gyrus and the left LBA to the frontal pole. Within INT, FC to the frontal pole and postcentral gyrus changed over time while changes in FC of the right LBA were also linked to symptom change. No significant interactions were observed when considering FC from CMA bilateral seeds or within ICA-derived networks.Results in this cohort suggest divergent longitudinal development of FC from bilateral LBA subregions in adolescents with internalizing disorders compared to healthy peers, possibly reflecting nonspecific treatment effects. Moreover, associations were found with symptom change. These results highlight the importance of differentiation of amygdala subregions in neuroimaging research in adolescents.

Abstract P212: Microinjection of the novel angiotensin, alamandine-(1-5) into insular cortex, increase blood pressure

It is well known that the renin-angiotensin system plays a critical role in cardiovascular control and hydro-electrolyte homeostasis. At least in some circumstances Alamandine (Ala) and Angiotensin-(1-7) can have selective actions such as reported in the insular córtex (IC). Hydrolysis of Angiotensin-(1-7) by angiotensin converting enzyme (ACE) can form the heptapeptide Angiotensin-(1-5) and recently our group demonstrated that hydrolysis of Ala by ACE can form Ala-(1-5). In this study we aimed to evaluate the cardiovascular effects produced by microinjection of Ala-(1-5) and Angiotensin-(1-5) into the rostral region of the posterior IC. Urethane (1.4g/kg) anesthetized rats were instrumented for measurement of mean arterial pressure (MAP), heart rate (HR) and renal sympathetic nerve activity (RSNA). Unilateral microinjection of vehicle (0,9% NaCl, 100nL, n=4), Ala (40pmol/100nl, n=7), Ala-(1-5) (400pmol/100nl, n=7) or Angiotensin-(1-5) (400pmol/100nl, n=5) were made into IC (+1.5mm rostral to the bregma). At the baseline, there was no difference between the groups (Vehicle: 85±2 mmHg and 388±5bpm, Ala: 83±9mmHg and 389±10bpm, Ala-(1–5): 91±6mmHg and 386±7bpm or Ang-(1-5): 96±9mmHg and 375±6bpm). Vehicle microinjection was performed as a control for nonspecific effects (ΔMAP:2±1mmHg, ΔHR:4±2bpm, ΔRSNA:3±1%). Ala microinjection into IC, confirmed our previous data showing a long-lasting (18±3min) increase in MAP (Δ=14±2mmHg), and this effect was associated with a significant increase in HR (Δ=32±4bpm) and in RSNA (Δ=36±6%). These responses were elicited immediately after Ala microinjection. When compared with vehicle and, very different from what was observed with Ala, microinjection of Ala-(1–5) evoked a significant increase in blood pressure without altering RSNA or HR (ΔMAP:22±2mmHg, ΔHR:-1±3bpm, ΔRSNA:1±3%). Differently from Ala evoked responses, the effects of Ala-(1–5) had a shorter duration (5.88±1.9min) without delay to beginning the effects. On other hand, microinjection of Angiotensin-(1-5) did not elicited autonomic and cardiovascular effects (ΔMAP:-1±3mmHg, ΔHR:-2±2bpm, ΔRSNA:2±3%). In conclusion, these data showed that the IC is a brain site that has unique effects of Ala (increase in MAP, HR and RSNA) and Ala-(1-5) (increase in MAP only) contrasting with the essentially absent effects of Angiotensin-(1-7) and Angiotensin-(1-5). These findings will contribute to extend our understanding of the brain renin-angiotensin system.

Reproducing the Solvatochromism of Merocyanines by PCM Calculations.

Polarizable continuum methods (PCM) have been widely employed for simulating solvent effects, in spite of the fact that they either ignore specific interactions in solution or only partially reproduce non-specific contributions. Examples of three solvatochromic dyes with a negative, a positive and a reverse behavior illustrate the achievements and shortcomings of PCM calculations and the causes for their variable success. Even when qualitatively mimicking non-specific solvent effects, departures of calculated values from experimental data may be significant (20-30%). In addition, they can utterly fail to reproduce an inverted behavior that is caused by significant specific contributions by the solvent. As shown through a theoretical model that rationalizes and predicts the solvatochromism of phenolate merocyanines based on DFT (Density Functional Theory) descriptors in the gas phase, PCM shortcomings are to be held responsible for its eventual failure to reproduce experimental data in solution.

Impact of Oil-in-Water Adjuvanted β-Glucan on Innate Immune Memory in Piglets.

The non-specific protective effects offered by the concept of "innate immune memory" might represent a promising strategy to tackle early-life threatening infections. Here we tested the potential of an in vitro selected β-glucan in inducing trained immunity using an in vivo porcine model. We assessed the leukocyte transcriptome using blood transcriptomic module (BTM), proinflammatory cytokines, and clinical scoring after a first "training" and a second "stimulation" phase. The possible induction of innate immune memory was tested during a "stimulation" by an LPS-adjuvanted Mycoplasma hyopneumoniae vaccine (Hyogen®) one day after weaning. Following the "training", no major group differences were found, with the exception of a plasma TNF that was only induced by Adj and Adj_BG treatment. After vaccination, all groups developed similar antibody responses. A significant induction of plasma TNF and IL-1β was found in groups that received Adj and Adj_BG. However, following vaccination, the expected early innate BTMs were only induced by the PBS group. In conclusion, the adjuvant alone, adjuvant-formulated β-glucan, or orally applied β-glucan were unable to enhance innate immune reactivity but rather appeared to promote innate immune tolerance. Such an immune status could have both positive and negative implications during this phase of the piglet's life.

Nonclinical Pharmacokinetics Study of OLX702A-075-16, N-Acetylgalactosamine Conjugated Asymmetric Small Interfering RNA (GalNAc-asiRNA).

In this study, the nonclinical pharmacokinetics of OLX702A-075-16, an RNA interference therapeutic currently in development, were investigated. OLX702A-075-16 is a novel N-acetylgalactosamine conjugated asymmetric small-interfering RNA (GalNAc-asiRNA) used for the treatment of an undisclosed liver disease. Its unique 16/21-mer asymmetric structure reduces nonspecific off-target effects without compromising efficacy. We investigated the plasma concentration, tissue distribution, metabolism, and renal excretion of OLX702A-075-16 following a subcutaneous administration in mice and rats. For bioanalysis, high-performance liquid chromatography with fluorescence detection was used. The results showed rapid clearance from plasma (0.5 to 1.5 hours of half-life) and predominant distribution to the liver and/or kidney. Less than 1% of the liver concentration of OLX702A-075-16 was detected in the other tissues. Metabolite profiling using liquid chromatography coupled with high-resolution mass spectrometry revealed that the intact duplex OLX702A-075-16 was the major compound in plasma. The GalNAc moiety was predominantly metabolized from the sense strand in the liver, with the unconjugated sense strand of OLX702A-075-16 accounting for more than 95% of the total exposure in the rat liver. Meanwhile, the antisense strand was metabolized by the sequential loss of nucleotides from the 3'-terminus by exonuclease, with the rat liver samples yielding the most diverse truncated forms of metabolites. Urinary excretion over 96 hours was less than 1% of the administered dose in rats. High plasma protein binding of OLX702A-075-16 likely inhibited its clearance through renal filtration. SIGNIFICANCE STATEMENT: This study presents the first comprehensive characterization of the in vivo pharmacokinetics of GalNAc-asiRNA. The pharmacokinetic insights gained from this research will aid in understanding toxicology and efficacy, optimizing delivery platforms, and improving the predictive power of preclinical species data for human applications.

Hubungan Antara Stres Kerja dan Keluhan Nyeri Punggung Bawah Pekerja Las Industri Pembuatan Kapal

Low Back Pain (LBP) is a widespread health issue that profoundly impacts workers' productivity and quality of life. In addition to non-specific health effects, LBP can result in long-term disability and considerable economic losses. Globally, LBP affects 619 million individuals, with projections suggesting an increase to 843 million by 2050, particularly Low Middle Income Countries (LMICs) in Asia and Africa. The shipbuilding industry, known for its physically strenuous tasks and stringent deadlines, presents unique ergonomic challenges that can exacerbate job-related stress and precipitate the onset of LBP. This study examines Project X PT Y, a pivotal entity in Indonesia's shipbuilding sector. Utilizing a quantitative methodology and a cross-sectional research design, the study encompasses all 26 workers from the welding workshop at Project X PT Y. The analysis, conducted using the Spearman test, reveals a robust monotonic relationship between work-related stress and LBP complaints, indicating that the incidence of LBP increases concomitantly with elevated work stress.

Placebo-related improvement with methylphenidate treatment in children with ADHD.

Non-specific effects of methylphenidate treatment, including expectancy and regression to the mean effects, contribute to the overall effect of methylphenidate on attention-deficit/hyperactivity disorder (ADHD) symptoms. Knowledge on the extent to which non-specific effects contribute to the overall effect and whether regression to the mean explains part of the non-specific effects, is currently lacking. A double-blind, randomized, placebo-controlled, cross-over trial was used to compare parent and teacher ratings of child ADHD symptoms at baseline and during treatment with placebo and 5, 10, 15 and 20mg of methylphenidate, twice daily. Participants were 5-13-year-old children with a DSM-5 diagnosis of ADHD (N = 45). The extent to which non-specific effects contributed to the effects of methylphenidate was determined by ADHD symptom reductions observed with placebo versus reductions observed with active doses of methylphenidate. The influence of regression to the mean was examined by estimating the contribution of baseline ADHD symptom severity to the effects observed with placebo treatment. Data were analyzed using multilevel analyses. We observed significant non-specific effects of methylphenidate for parent-rated ADHD symptoms, but not for teacher-rated symptoms. For parent reported hyperactive/impulsive symptoms, higher baseline symptoms predicted larger effects with placebo, indicating regression to the mean effects. For parent-reports, a significant part of the overall effect of methylphenidate treatment is explained by non-specific effects. Our findings stress the importance of taking non-specific effects into account when evaluating methylphenidate treatment, by including teacher-reports and using a double baseline assessment during titration. Comparing active medication with a placebo in the titration trial has the potential to identify non-specific effects.

The ‘ABC’ of split-nanoluciferase HIF heterodimerization bioassays: Applications, Benefits & Considerations

Hypoxia-inducible factors (HIF) are interesting targets for multiple therapeutic indications. While HIF activation is desired for the treatment of anemia-related and ischemic diseases, HIF inhibition is of tremendous interest to anti-cancer drug development. Different signaling events within the HIF pathway are being targeted by drug discovery programs, with a special interest in HIF-selective (possibly also HIF1/2 isoform-selective) compounds. In this study, we applied recently developed cell-based split-nanoluciferase HIF heterodimerization assays to study the effects of compounds, targeting HIF activity by various mechanisms of action. This study shows that the application of similar or diverse assay protocols allows to detect various influences on HIF heterodimerization as a key signaling event in the oxygen sensing pathway: increased HIF heterodimerization (roxadustat, MG-132), decreased HIF heterodimerization (PX-478, ibuprofen) and direct (HIF isoform-selective) heterodimerization inhibiting effects (PT-2385). Changes in treatment time and in the assay protocol allowed to assess direct and indirect effects on HIFα-HIFβ heterodimerization. In addition to the evaluation of applications of these new bioassays regarding pharmacological characterizations, benefits and considerations are discussed related to the use of cellular, luminescent-based bioassays. Briefly, benefits include the bidirectional nature of the biological readout, the upstream mechanism of detection, the differentiation between HIF1 and HIF2 effects and the simulation of various conditions. Specific and general considerations include cell-based, technical and disease/drug-related aspects (e.g., non-specific effects, color interference). In summary, the versatility of these bioassays offers benefits in widespread applications regarding drug discovery and pharmacological characterization of various therapeutics, applying either the same or optimized experimental protocols.

DNA methylation correlates of chronological age in diverse human tissue types

BackgroundWhile the association of chronological age with DNA methylation (DNAm) in whole blood has been extensively studied, the tissue-specificity of age-related DNAm changes remains an active area of research. Studies investigating the association of age with DNAm in tissues such as brain, skin, immune cells, fat, and liver have identified tissue-specific and non-specific effects, thus, motivating additional studies of diverse human tissue and cell types.ResultsHere, we performed an epigenome-wide association study, leveraging DNAm data (Illumina EPIC array) from 961 tissue samples representing 9 tissue types (breast, lung, colon, ovary, prostate, skeletal muscle, testis, whole blood, and kidney) from the Genotype-Tissue Expression (GTEx) project. We identified age-associated CpG sites (false discovery rate < 0.05) in 8 tissues (all except skeletal muscle, n = 47). This included 162,002 unique hypermethylated and 90,626 hypomethylated CpG sites across all tissue types, with 130,137 (80%) hypermethylated CpGs and 74,703 (82%) hypomethylated CpG sites observed in a single tissue type. While the majority of age-associated CpG sites appeared tissue-specific, the patterns of enrichment among genomic features, such as chromatin states and CpG islands, were similar across most tissues, suggesting common mechanisms underlying cellular aging. Consistent with previous findings, we observed that hypermethylated CpG sites are enriched in regions with repressed polycomb signatures and CpG islands, while hypomethylated CpG sites preferentially occurred in non-CpG islands and enhancers. To gain insights into the functional effects of age-related DNAm changes, we assessed the correlation between DNAm and local gene expression changes to identify age-related expression quantitative trait methylation (age-eQTMs). We identified several age-eQTMs present in multiple tissue-types, including in the CDKN2A, HENMT1, and VCWE regions.ConclusionOverall, our findings will aid future efforts to develop biomarkers of aging and understand mechanisms of aging in diverse human tissue types.

Exploring the relationship between gut microbiota and breast cancer risk in European and East Asian populations using Mendelian randomization

BackgroundSeveral studies have explored the potential link between gut microbiota and breast cancer; nevertheless, the causal relationship between gut microbiota and breast cancer remains unclear.MethodsWe utilized summary statistics from genome-wide association studies (GWAS) of the gut microbiome from the MiBioGen project with summary data from GWAS on breast cancer from the FinnGen consortium and the IEU database, with the IEU data sourced from the Biobank Japan. Preliminary statistical analyses were conducted using inverse variance weighting (IVW), supplemented by various sensitivity analysis methods, including MR-Egger regression, weighted median, weighted mode, simple median, and simple mode, to ensure the robustness of our findings. Heterogeneity and pleiotropy were assessed to avoid misleading conclusions caused by unconsidered confounders or non-specific effects of genetic variants, ensuring that the results reflect a genuine causal relationship.ResultsIn European populations, four types of gut microbiota were associated with breast cancer. The genus Erysipelatoclostridium was positively associated with the risk of breast cancer, with an odds ratio (OR) of 1.21 (95% confidence interval [CI] 1.083–1.358), false discovery rate (FDR) = 0.0039. The class Coriobacteriia, order Coriobacteriales, and family Coriobacteriaceae, which belong to the same phylogenetic system, showed a consistent inversely association with breast cancer risk, with an OR of 0.757 (95% CI 0.616–0.930), FDR = 0.0281. In East Asian populations, three types of gut microbiota were related to breast cancer. The Eubacterium ruminantium group was positively associated with breast cancer risk, with an OR of 1.259 (95% CI 1.056–1.499), FDR = 0.0497. The families Porphyromonadaceae and Ruminococcaceae were inversely associated with breast cancer risk, with ORs of 0.304 (95% CI 0.155–0.596), FDR = 0.0005, and 0.674 (95% CI 0.508–0.895), FDR = 0.03173, respectively. However, these two taxa had limited instrumental variables, restricting the statistical power and potentially affecting the interpretation of the results.ConclusionThis MR analysis demonstrated a probable causal link between specific gut microbiota and breast cancer. This study, through Mendelian randomization analysis comparing European and East Asian populations, reveals that gut microbiota may influence breast cancer risk differently across populations, providing potential directions for developing targeted prevention and treatment methods.

Known and unknown hymecromone. A review

Hymecromone (4-MU) is a recognized agent currently used in clinical practice. Since 1960, hymecromone has been used in many countries as a choleretic and cholespasmolytic, a drug approved for use in humans with biliary tract disorders. The review presents both traditional European and Russian studies of the selective antispasmodic and choleretic properties of hymecromone, due to which hymecromone is the drug of choice for the treatment of biliary tract diseases, as well as new fundamental and clinical studies of numerous pleiotropic effects of 4-MU associated with inhibition of hyaluronic acid and many other properties of this exciting molecule. These include antibacterial, antiviral, and nonspecific anti-inflammatory effects. Positive results have been demonstrated in carbohydrate and lipid metabolism disorders, autoimmune diseases, as well as liver, heart, and kidney diseases. Numerous in vitro and in vivo studies have been presented in pancreatic, prostate, skin, esophagus, breast, liver, ovary, bone cancers, metastatic lesions, leukemia, autoimmune and inflammatory diseases. Hymecromone is indicated not only as a choleretic and cholespasmolytic but also as a choleseptic in cholangitis and chronic cholecystitis, including opisthorchiasis, which does not disagree with its label. Odecromone® (hymecromone, tablets 200 mg) is available on the Russian market; it replaced the originator drug and is its fully equivalent generic.