Early Marker Research Articles

Early markers of induced pluripotent stem cell hematopoietic development

The generation of immune cell populations from induced pluripotent stem cells (iPSCs) is a valuable model to study mechanisms that control hematopoietic development; it also is a promising approach to develop immunotherapeutic strategies to treat various diseases, including hereditary, oncological and infectious ones. To date, it has been demonstrated that iPSCs can differentiate into different immune cells, including macrophages, neutrophils, natural killer cells and T cells. However, the protocols suggested so far are experimental, and they require optimization, standardization and scaling. Solution to these tasks requires methods allowing to predict the efficacy of ongoing differentiation at early differentiation stages. Here, we evaluated whether iPSC hematopoietic/myeloid differentiation can be monitored by means of flow cytometry. Human iPSCs were differentiated into hematopoietic/myeloid cells using two protocols previously suggested for the generation of macrophages from iPSCs. The protocols differed by methods used to induce early and late stages of cell differentiation. At early differentiation stages, the protocols differed by approaches used to induce the generation of mesoderm and hemogenic endothelium, i.e., 2D differentiation in the presence of exogenous factors known to promote mesoderm and hemogenic endothelium development (“factor-dependent” protocol) or 3D differentiation in the absence of exogenous cytokines and growth factors (“spontaneous” protocol). At late differentiation stages, the protocols differed by factors added to the cultures to promote hematopoietic/myeloid specification (i.e., SCF, FGF2, IL-6, IL-3 and M-CSF or only IL-3 and M-CSF). At different stages of differentiation, the expressions of antigens known to be expressed by mesoderm, hemogenic endothelium and hematopoietic cells (i.e., CD309, CD34, CD31, CD43 and CD45) were evaluated. At early differentiation stages, the main phenotypic changes observed in cell cultures were an upregulation of the expression of CD309 (a receptor for vascular endothelial growth factor), the appearance of the expression of sialophorin CD43 and the expression of CD34 antigen. In cells cultured in 2D factor-dependent conditions, these changes appeared earlier and were more pronounced as compared with cells cultured in 3D “spontaneous” conditions. The results suggest that CD309 and/or CD43 are valuable markers for an early prediction of the effectiveness of iPSC differentiation into hematopoietic/ myeloid progeny.

Enteric Nervous System and Its Relationship with Neurological Diseases.

The enteric nervous system (ENS) is a fundamental component of the gastrointestinal system, composed of a vast network of neurons and glial cells. It operates autonomously but is interconnected with the central nervous system (CNS) through the vagus nerve. This communication, known as the gut-brain axis, influences the bidirectional communication between the brain and the gut. Background/Objectives: This study aimed to review neurological pathologies related to the ENS. Methods: To this end, a comprehensive literature search was conducted in the "PubMed" database. Articles available in "free format" were selected, applying the filters "Humans" and limiting the search to publications from the last ten years. Results: The ENS has been linked to various neurological diseases, from autism spectrum disorder to Parkinson's disease including neurological infection with the varicella zoster virus (VZV), even sharing pathologies with the CNS. This finding suggests that the ENS could serve as an early diagnostic marker or therapeutic target for neurological diseases. Gastrointestinal symptoms often precede CNS symptoms, and the ENS's accessibility aids in diagnosis and treatment. Parkinson's patients may show intestinal lesions up to twenty years before CNS symptoms, underscoring the potential for early diagnosis. However, challenges include developing standardized diagnostic protocols and the uneven distribution of dopaminergic neurons in the ENS. Continued research is needed to explore the ENS's potential in improving disease prognosis. Conclusions: The ENS is a promising area for early diagnosis and therapeutic development. Nevertheless, it is essential to continue research in this area, especially to gain a deeper understanding of its organization, function, and regenerative capacity.

Serum exosomal miR-200c is a potential diagnostic biomarker for breast cancer

Background Breast cancer (BC) is one of the most common malignancies in women. Exosomes are widely found in body fluids and carry microRNAs (miRNAs) that reflect the biological properties of the parental cells. Our study aimed to investigate the differential expression of miR-200c in BC serum exosomes and its diagnostic value. Methodology miRNA profiles in culture supernatant exosomes of normal mammary epithelial cells MCF-10A and BC cells (MCF-7, MDA-MB-231, MCF-7 Taxol) were examined by miRNA deep sequencing to screen for significantly differentially expressed miRNAs; Transmission electron microscopy (TEM), Nanoparticle tracking analysis (NTA), and Western blot were used to identify exosomes; qPCR was used to detect the expression level of miR-200c in cellular exosomes and serum exosomes; The efficacy of individual and combined tests of each indicator to diagnose BC was evaluated using receiver operating characteristic (ROC) curves. Results We identified typical exosome features by TEM, NTA and Western blot, indicating successful exosome extraction. Then our miRNA sequencing results and qRT-PCR experiments showed that miR-200c was significantly down-regulated in BC cell exosomes. In addition, we divided the clinical serum samples into two cohorts according to region, and in independent cohort I, the serum exosomal miR-200c levels of BC patients were significantly lower than those of healthy controls. In cohort II, serum exosomal miR-200c expression was significantly lower in the BC group than in the control and benign breast disease (BBD) groups, whereas miR-200c expression in the BBD group was not statistically different from that in the control group. ROC analyses in both independent cohorts confirmed that serum exosomal miR-200c could differentiate between patients with and without BC disease and could be used as an early diagnostic marker for BC disease. Conclusion Serum exosome miR-200c can be used as a potential biomarker for the diagnosis of BC, and combined with conventional serum diagnostic markers AFP, CA125 and CA153 can help to improve diagnostic efficiency.

Lung-Kidney Axis in Cystic Fibrosis: Early Urinary Markers of Kidney Injury Correlate with Neutrophil Activation and Worse Lung Function.

Adult people with cystic fibrosis (PwCF) have a higher risk of end-stage kidney disease than the general population. The nature and mechanism of kidney disease in CF are unknown. This study quantifies urinary kidney injury markers and examines the hypothesis that neutrophil activation and lung infection are associated with early kidney injury in CF. Urinary total protein, albumin, and markers of kidney injury and neutrophil activation, normalized to creatinine, as well as urinary immune cells, were quantified in CF (n = 48) and healthy (n = 33) cohorts. Infection burden and chronicity were defined by sputum culture and serum titers of anti-bacterial antibodies. PwCF had increased urinary protein levels, consisting of low-molecular-weight tubular injury markers, independent of glomerular filtration rate (eGFR). This finding suggests subclinical renal injury processes. Urinary analysis of the CF cohort identified different associations of urinary injury markers with aminoglycoside exposure, lung function, and neutrophil activation. High urinary KIM-1 levels and increased prevalence of neutrophils among urine immune cells correlated with decreased lung function in PwCF. The relationship between tubular injury and decreased lung function was most prominent in patients harboring chronic Pseudomonas aeruginosa infection. Increased urinary tubular injury markers in PwCF suggest early subclinical renal injury not readily detected by eGFR. The strong association of high urinary KIM-1 and neutrophils with diminished lung function and high Pseudomonas aeruginosa burden suggests that pulmonary disease may contribute to renal injury in CF.

Impaired Balance Predicts Cardiovascular Disease in 70-Year-Old Individuals-An Observational Study From the Healthy Aging Initiative.

Limited research has explored balance problems as a prospective risk factor for cardiovascular disease (CVD). This study aimed to characterize the association between balance measures and the risk of incident CVD in a population of 70-year-olds. From 2012 to 2022 a cohort of 4927 older individuals who were CVD free underwent balance assessments using a balance board. Measurements included lateral and anterior-posterior sway, along with a safety limit of stability in a subcohort (N=2782). Time to first hospitalization for CVD, encompassing stroke, myocardial infarction, or angina pectoris was the primary outcome. Multivariable regression models assessed associations between balance parameters and CVD risk. Over a mean follow-up of 4.9 years, 320 individuals were hospitalized for CVD. In a balance test with eyes open, increased lateral sway at baseline was associated with a higher risk of CVD (hazard ratio [HR], 1.014 [95% CI, 1.004-1.025], P=0.005, per mm increased sway), after adjustment for traditional risk factors for CVD. Similarly, individuals with CVD during follow-up exhibited higher lateral sway with eyes closed at baseline (HR, 1.015 [95% CI, 1.005-1.025], P=0.002, per mm increased sway), after multivariable adjustment. The 4 strongest independent predictors of CVD included lateral sway and were associated with a population attributable fraction of 61% (95% CI, 54-68). In community-dwelling 70-year-olds, impaired lateral balance was an independent predictor of later CVD, after adjustment for traditional risk factors. This may suggest that position balance could be used as an early risk marker for underlying atherosclerotic disease.

Screening miRNAs to Hinder the Tumorigenesis of Renal Clear Cell Carcinoma Associated with KDR Expression.

This study delved into the role of Kinase Insert Domain Receptor (KDR) and its associated miRNAs in renal cell carcinoma through an extensive computational analysis. The potential of our findings to guide future research in this area is significant. Our methods, which included the use of UALCAN and GEPIA2 databases, as well as miRDB, MirDIP, miRNet v2.0, miRTargetLink, MiEAA v2.1, TarBase v8.0, INTERNET, and miRTarBass, were instrumental in identifying the regulation of miRNA associated with KDR expression. The predicted miRNA was validated with the TCGA-KIRC patients' samples by implementing CancerMIRNome. The TargetScanHuman v8.0 was implemented to identify the associations between human miRNAs and KDR. A Patch Dock server analyzed the interactions between hsa-miR-200b-3p-KDR and hsa-miR-200b-3p with KDR. The KDR expression rate was investigated in the Kidney Renal Cell Carcinoma (KIRC) samples, and adjacent normal tissues revealed that the expression rate was significantly higher than the normal samples, which was evident from the strong statistical significance (P = 1.63e-12). Likely, the KDR ex-pression rate was estimated as high at tumor grade 1 and gradually decreased till the metastasis grade, reducing the survival rate of the KIRC patients. To identify these signals early, we predicted a miRNA that could trigger the expression of KDR. Furthermore, we uncovered the potential associations between miR-200c-3p expressions by regulating KDR towards the progression of KIRC. Upon examining the outcome, it became evident that miR-200c-3p was significantly down-regulated in KIRC compared to the normal samples. Moreover, the negative correlation was obtained for hsa-miR-200c-3p (R = - 0.276) along with the KDR expression describing that the increased rate of hsa-miR-200c-3p might reduce the KDR expression rate, which may suppress the KIRC initiation or progres-sion. The in-silico analysis indicated that the significant increase in KDR expression during the initiation of KIRC could serve as an early diagnostic marker. Moreover, KDR could be utilized to identify advancements in KIRC stages. Additionally, hsa-miR-200c-3p was identified as a potential regulator capable of downregulating and upregulating KDR expression among the 24 miRNAs screened. This find-ing holds promise for future research endeavors. Concurrent administration of the FDA-approved 5-fluor-ouracil with KIRC drugs, such as sorafenib, zidovudine, and everolimus, may have the potential to en-hance the therapeutic efficacy in downregulating hsa-miR-200c-3p. However, further in vitro studies are imperative to validate these findings and gain a comprehensive understanding of the intricate regulatory interplay involving hsa-miR-200c-3p, KDR, 5-fluorouracil, and other FDA-approved drugs for the treat-ment of KIRC. This will facilitate the identification of KIRC stage progression and its underlying pre-ventative mechanisms.

Cisplatin-Induced Metabolic Responses Measured with Raman Spectroscopy in Cancer Cells, Spheroids, and Canine-Derived Organoids.

Ex vivo assessment of drug response with conventional cell viability assays remains the standard practice for guiding initial therapeutic choices. However, such ensemble approaches fail to capture heterogeneities in treatment response and cannot identify early markers of response. Here, we leverage Raman spectroscopy (RS) as an accurate, low-cost, extraction-free, and label-free approach to track metabolic changes in cancer cells, spheroids, and organoids in response to cisplatin treatment. We identified 12 statistically significant metabolites in cells and 19 metabolites in spheroids and organoids as a function of depth. We show that the cisplatin treatment of 4T1 cells and spheroids results in a shift in metabolite levels; metabolites including nucleic acids such as DNA, 783 cm-1 with p = 0.00021 for cells; p = 0.02173 for spheroids, major amino acids such as threonine, 1338 cm-1 with p = 0.00045 for cells; p = 0.01022 for spheroids, proteins such as amide III, 1248 cm-1 with p = 0.00606 for cells; p = 0.00511 for spheroids serve as early predictors of response. Our RS findings were also applicable to canine-derived organoids, showing spatial variations in metabolic changes as a function of organoid depth in response to cisplatin. Further, the metabolic pathways such as tricarboxylic acid (TCA)/citric acid cycle and glyoxylate and dicarboxylate metabolism that drive drug response showed significant differences based on organoid depth, replicating the heterogeneous treatment response seen in solid tumors where there is a difference from the periphery to the tumor core. Our study showcases the versatility of RS as a predictive tool for treatment response applicable from cells to organotypic cultures, that has the potential to decrease animal burden and readout time for preclinical drug efficacy.

Analysis of Intact In Vivo Peroxidized Phospholipids from Bovine Retina via LC-MS/MS and GC-MS/MS Using Autoxidized Retina Reference Standards.

Highly unsaturated fatty acids (HUFAs) are vulnerable to oxygen attack, thus making HUFA-rich, high metabolic rate/reactive oxygen species (ROS)-generating neurological tissue particularly susceptible to increased oxidative stress. Lipid oxidation is a putative early stage marker of neurodegenerative diseases, suggesting that reliable monitoring of oxidized neural lipids in vivo reveals early pathogenesis. Here, we present a novel methodology to detect and quantify intactin vivo ROS-driven peroxidized phospholipids (LPOx-PLs) in bovine retina extract. A protocol for preparing autoxidized pure phospholipids (PLs) and complex retinal extracts served as reference standards and was adapted to enable analytical parameter development. Fatty acid profiles of bovine retinas were first established with routine gas chromatography (GC) methods and used to customize mass spectrometry scanning for major HUFA-carrying PLs in the retinal extract. Targeted multiple reaction monitoring (MRM) scanning via triple quadrupole tandem mass spectrometry detected native (unoxidized) and oxidation-damaged PL regardless of the position of the O or O2 addition along the acyl chains and enabled quantification of relative signals from intact native and oxidized PL (5%-10% CV). MRM-triggered information-dependent acquisition (IDA) spectra confirmed the structure of peroxidized PLs, revealing that peroxidized species (+O-OH) dominated over single O-added species in vitro and in vivo. Positive identification and relative quantification are reported for 12 selected in vivo native and peroxidized phosphatidylcholines and phosphatidylethanolamines. These results enable future studies of the initial peroxidation due to toxins, genetics, or other initiating events influencing in vivo oxidation levels and potentially the effectiveness of strategies to mitigate this mechanism of action.

Prognostic Significance of Lactate Clearance in Cardiogenic Pulmonary Edema in the Emergency Department.

Background and Objectives: Acute cardiorespiratory failure disrupts the delicate balance of energy supply, demand, and consumption, with elevated lactate levels and decreased blood pH serving as crucial indicators. Acute cardiogenic pulmonary edema (ACPO), a common cause of acute respiratory failure, poses a substantial mortality risk. Lactate, a byproduct of pyruvate reduction, is a pertinent marker in perfusion assessment. Lactate clearance (LC) has proven prognostic efficacy in various conditions but lacks consensus on its predictive power in acute cardiogenic pulmonary edema. Materials and Methods: This prospective observational study, conducted in a metropolitan area's third-level emergency department, involved patients with cardiogenic pulmonary edema from May 2021 to August 2023. The inclusion criteria specified acute cardiogenic pulmonary edema, excluding patients with incomplete data or other respiratory conditions. Lactate clearance, calculated at presentation and after 6 h, served as the primary outcome predictor. Our data analysis employed logistic regression, the ROC curve, and statistical tests. Results: The cohort of 106 patients revealed that a lactate clearance below 14.29% was significantly associated with mortality. While 51.6% of survivors were discharged, LC's predictive success for discharge was inconclusive. Logistic regression underscored the significance of lactate clearance, with a one-unit increase yielding a 5.55-fold probability of survival. The AUC for LC was 0.759. Conclusions: This study pioneers the exploration of lactate clearance in patients with acute cardiogenic pulmonary edema. LC below 14.29% signifies a poor prognosis, emphasizing its potential as an early treatment initiation marker. While acknowledging this study's limitations, we advocate for further multicenter research to refine the understanding of lactate clearance in this context.

Diabetic Kidney Disease: Contribution of Phenyl Sulfate Derived from Dietary Tyrosine upon Gut Microbiota Catabolism.

Deranged gut microbiota can release increased levels of uremic toxins leading to exacerbated kidney injury. In diabetic kidney disease (DKD), phenyl sulfate (PS) derived from tyrosine catabolism by gut microbiota has been demonstrated to be both an early diagnostic marker and a therapeutic target. In this perspective article, we summarize PS generation pathways and recent findings on PS and kidney injury in DKD. Increasing evidence has shown that the underlying mechanisms of PS-induced kidney injury mainly involve oxidative stress, redox imbalance, and mitochondrial dysfunction, which all may be targeted to attenuate PS-induced kidney injury. For future research directions, we think that a deeper understanding of the pathogenic role of PS in kidney injury using a variety of diabetic animal models should be investigated. Moreover, we also suggest beneficial approaches that could be used to mitigate the deleterious effect of PS on the kidney. These approaches include caloric restriction, tyrosine restriction, and administration of ketogenic drugs, ketogenic diets or natural products; all of which should be conducted under obese and diabetic conditions.

Analysis of laboratory and instrumental data in young patients with COVID-19 and cardiovascular disorders

Introduction. Despite significant information on cardiovascular damage in older patients with COVID- 19, there is no clear understanding of the variability of cardiac diseases manifestations in young patients. Additionally, summarized data characterizing the severity of myocardial damage in these patients is lacking.Aim. To compare laboratory and functional study data and identify predictors of cardiovascular damage in young patients with COVID-19.Materials and methods. An analysis of 4,453 medical records from 2020 to 2022 was conducted. The focus was on patient age (18- 44 years), primary diagnosis, severity of COVID-19, comorbid conditions, timing of cardiovascular symptom onset, and laboratory and functional study data characterizing cardiovascular system damage. According to the inclusion and exclusion criteria, 132 medical records of patients aged 18 to 44 years with moderate COVID-19 were selected, including 49 with cardiovascular issues and 83 without cardiovascular pathology.Results. Comparative analysis showed that cardiovascular system damage in young patients with moderate COVID-19, compared to those without cardiovascular pathology, was associated with elevated levels of C-reactive protein (CRP) (p=0.001), brain natriuretic peptide (NTproBNP) (p<0.0001), D-dimer (p<0.0001), and troponin I (p<0.0001). CRP levels in patients with pericarditis and acute myocardial infarction (AMI) were higher than in those with acute cerebrovascular accident (ACVA) (p=0.001). No differences in D-dimer levels were found between subgroups (p>0.05). Troponin I and NTproBNP levels in the AMI subgroup were higher than in the pericarditis (p<0.0001 and p=0.047, respectively) and ACVA (p=0.001 and p=0.043, respectively) subgroups. Patients with cardiovascular disorders had a reduced left ventricular ejection fraction (LVEF) (p=0.005) without significant changes in left ventricular end-systolic (p=0.200) and end-diastolic volumes (p=0.119). The lowest LVEF was found in the AMI and ACVA subgroups (p=0.001). Correlation-regression analysis revealed associations between cardiovascular complications and NTproBNP (r=0.673, p<0.001), troponin I (r=0.543, p<0.001), and D-dimer (r=0.363, p<0.001), as well as LVEF (r=-0.341, p<0.001). Significant stochastic relationships were also found between D-dimer and troponin I (r=0.532, p<0.001), NTproBNP (r=0.545, p<0.001); and between LVEF and troponin I (r=-0.420, p<0.001), NTproBNP (r=-0.314, p<0.001). Multivariate regression analysis showed that NTproBNP, with a sensitivity of 86.5% and specificity of 81.9%, is an independent predictor of cardiovascular complications in young patients with moderate COVID-19 (OR=1.175, p=0.001).Conclusion. The results indicate that the severity of COVID-19, regardless of patient age and comorbidities, can be a factor in the development of cardiovascular complications. NTproBNP levels in the blood of COVID-19 patients can be an early marker of cardiovascular risk. However, further research is needed to confirm the pathogenetic role of cardiotropic protein in the development of pericarditis, AMI, and ACVA in these patients.

A - 21 The Association between Tau Pathology and Cognitive Impairment in Alzheimer’s Disease in the Democratic Republic of Congo

Abstract Objective Previous studies in Western countries have shown that tau accumulation is more closely connected with cognitive decline in Alzheimer’s disease (ad). This association has not yet been investigated in the Sub-Saharan African countries/Democratic Republic of Congo (DRC). We investigated the association between phosphorylated tau (p-Tau) level and cognitive performance of older adults from the DRC. Method Blood and neuropsychological tests were collected on 80 Congolese older adults with suspected ad. Participants were classified according to the biomarkers of their ATN profiles with positive “A” values based on β-amyloid <0.061 and “T” values based on p-Tau 181 < 4.50, as well as APOEε4 using Simoa assays. An analysis of covariance examined the differences in memory performance for each pathological group, controlling for age, education, gender, and APOEε4 status. Results A-T- (n = 36) consisted of age (M = 71.3, SD = 7.44), gender (48.6% female), education level (M = 9.80, SD = 5.44), and APOEε4 (38.2%). A + T- (n = 29) consisted of age (M = 73.1, SD = 7.75), gender (62.1% female), education level (M = 7.21, SD = 5.92), and APOEε4 (41.4%). A + T+ (n = 15) consisted of age (M = 78.1, SD = 4.62), gender (60.0% female), education level (M = 6.60, SD = 3.46), and APOEε4 (93.3%). Overall, there were significant effects of memory performances on A-T- participants: verbal memory immediate (p = 0.0089), delayed (p = 0.00096), and recognition (p = 0.0104); and visual memory delayed (p = 0.00058) and recognition (p = 0.00039). Conclusion Reduced memory performance and p-Tau levels were associated in Congolese adults with and without ad. This study suggests abnormal tau accumulation underlies cognitive deficits in ad and could be used as an early marker of ad pathology for various populations in Sub-Saharan African countries.

REM sleep without atonia and neurocognitive function in isolated REM sleep behaviour disorder: Cross-sectional and longitudinal study.

This study investigated the relationship between rapid eye movement sleep without atonia and cognitive profiles in individuals diagnosed with isolated rapid eye movement sleep behaviour disorder, assesssing both cross-sectional associations and their link to phenoconversion in a longitudinal follow-up. Participants underwent video-polysomnography, neurological examination, neuropsychological tests and structured interviews to confirm isolated rapid eye movement sleep behaviour disorder. Rapid eye movement sleep without atonia was manually scored using the Montreal method, and participants were categorized into either high or low electromyography activity groups, based on their tonic and phasic electromyography activities. The cross-sectional study included 250 patients with isolated rapid eye movement sleep behaviour disorder, revealing that those with high tonic electromyography activity exhibited significantly lower scores in the constructional praxis recall than those with low tonic electromyography activity (p = 0.002). In the longitudinal study, 79 participants (63 isolated rapid eye movement sleep behaviour disorder and 16 phenoconversion), tracked for at least 5 years, demonstrated that high tonic electromyography activity (odds ratio: 6.14; 95% confidence interval: 1.23-30.60; p = 0.027) and lower performance on the Trail Making Test A (odds ratio: 0.23; 95% confidence interval: 0.11-0.70; p = 0.007) were associated with future phenoconversion. These results confirm the link between tonic electromyography activity and neurodegeneration in isolated rapid eye movement sleep behaviour disorder. Combining rapid eye movement sleep without atonia assessment with cognitive evaluation could serve as an early predictive marker for phenoconversion in clinical settings.

Evolving cartilage strain with pain progression and gait: a longitudinal study post-ACL reconstruction at six and twelve months.

Anterior cruciate ligament (ACL) injuries are prevalent musculoskeletal conditions often resulting in long-term degenerative outcomes such as osteoarthritis (OA). Despite surgical advances in ACL reconstruction, a significant number of patients develop OA within ten years post-surgery, providing a patient population that may present early markers of cartilage degeneration detectable using noninvasive imaging. This study aims to investigate the temporal evolution of cartilage strain and relaxometry post-ACL reconstruction using displacement under applied loading MRI and quantitative MRI. Specifically, we examined the correlations between MRI metrics and pain, as well as knee loading patterns during gait, to identify early candidate markers of cartilage degeneration. Twenty-five participants (female/male = 15/10; average age = 25.6 yrs) undergoing ACL reconstruction were enrolled in a prospective longitudinal cohort study between 2022 and 2023. MRI scans were conducted at 6- and 12-months post-surgery, assessing T2, T2*, and T1ρ relaxometry values, and intratissue cartilage strain. Changes in pain were evaluated using standard outcome scores, and gait analysis assessed the knee adduction moment (KAM). Regressions were performed to evaluate relationships between MRI metrics in cartilage contact regions, patient-reported pain, and knee loading metrics. Increases in axial and transverse strains in the tibial cartilage were significantly correlated with increased pain, while decreases in shear strain were associated with increased pain. Changes in strain metrics were also significantly related to KAM at12 months. Changes in cartilage strain and relaxometry are related to heightened pain and altered knee loading patterns, indicating potential early markers of osteoarthritis progression. These findings underscore the importance of using advanced MRI for early monitoring in ACL-reconstructed patients to optimize treatment outcomes, while also highlighting KAM as a modifiable intervention through gait retraining that may positively impact the evolution of cartilage health and patient pain. Increased axial and transverse strains in the tibial cartilage from 6 to 12 months post-ACL reconstruction were significantly correlated with increased pain, suggesting evolving changes in cartilage biomechanical properties over time.Decreases in shear strain in inner femoral and central tibial cartilage regions were linked to increased pain, indicating alterations in joint loading patterns.Decreases in shear strain in the inner femoral cartilage were significantly associated with decreased 12-month knee adduction moment (KAM), a surrogate for medial cartilage knee loading during walking.

The Effect of Upright Posture on Left Atrial Strain in Competitive Athletes.

Left atrial strain (LAS) assessment by speckle tracking echocardiography (STE) has been shown to be a remarkable means of quantifying LA function as an early marker of LV pathology. As exercise testing is also performed on a treadmill, the aim of this study was to investigate the effect of upright posture on LAS in healthy athletes. Fifty male athletes (mean age 25.7 ± 7.3 years) underwent transthoracic echocardiography (TTE) in the upright and left lateral positions. In addition to the conventional echocardiographic parameters, in all athletes, LA conduction strain (LAScd), contraction strain (LASct), reservoir strain (LASr), and maximum LA volume (LAVmax) were assessed by STE in both positions. Comparing upright posture and the left lateral position, LAScd (-14.0 ± 5.9% vs. -27.4 ± 7.1%; p < 0.001), LASct (-4.6 ± 3.5% vs. -11.3 ± 4.1%; p < 0.001), LASr (18.7 ± 7.6% vs. 38.7 ± 8.0%; p < 0.001), and LAVmax (24.4 ± 8.8% vs. 50.0 ± 14.2%) differed significantly. Upright posture has a significant effect on LA deformation, with decreased LAScd, LASct, and LASr. The results of this study contribute to the understanding of athletes' hearts and must be considered when performing echocardiography in healthy athletes on a treadmill.

Correlation of anogenital distance from childhood to age 9 years-a prospective population-based birth cohort-the Odense Child Cohort.

Does anogenital distance (AGD) - distance from the anus to the genitals - correlate from infancy (3 months) to the age of 9 years in boys and girls? In boys, AGD correlated from infancy to 9 years of age, whereas in girls, correlations were weaker, especially between infancy and later childhood. AGD is considered a marker for prenatal androgen action. In males, reduced AGD is associated with testicular cancer, infertility, and lower sperm count. In females, AGD is associated with endometriosis and polycystic ovary syndrome. In the Odense Child Cohort, a prospective population-based birth cohort, pregnant women were enrolled in early pregnancy. AGD and BMI were measured repeatedly in children at ages 3 and 18 months, as well as at 3, 5, 7, and 9 years. AGD was measured from the anus to the scrotum (AGDas) and to the penis (AGDap) in 1022 boys, and to the posterior fourchette and the clitoris in 887 girls repeatedly between the age of 3 months to 9 years. In total, 7706 assessments were made. AGD was adjusted for body weight, and SD scores (the difference between individual AGD and the mean of AGD in the population divided by SD of AGD) were calculated for each child. Pearson correlation coefficient (r) of each measurement was performed to investigate whether individual AGD was stable during childhood. Short predictive values at 3 months (20th percentile) to 9 years were investigated using the AUC produced by the receiver operating characteristic curve. In boys, AGD/body size-index SD score correlated significantly between infancy and 9 years, strongest for AGDas (r = 0.540 P > 0.001). In girls, weaker significant correlation coefficients were found between AGD at infancy and 9 years; higher correlation coefficients were found between AGD from 3 to 9 years (P > 0.001). Short AGDas in infancy predicted short AGDas in boys aged 9 years (AUC: 0.767, sensitivity 0.71, specificity 0.71). The predictive values of short infant AGDap, penile width (in boys), and AGD (in girls) concerning short outcomes at 9 years were low. The AGD measurements are less precisely measurable in girls compared to boys, especially in infancy, resulting in less reproducible measurements. Additionally, because AGD is shorter in girls, the same absolute measurement error is relatively more significant, potentially contributing to greater variability and lower reproducibility in girls. This may contribute to the weaker correlations in girls compared to boys. In boys, AGDas, relative to body size, correlated from infancy to 9 years, suggesting that AGD in infancy can be considered a non-invasive marker of later reproductive health. Further follow-up studies are needed to evaluate long-term individual tracking of AGD as well as assessment of childhood AGD as early marker of adult reproductive health. This study was supported by Odense University Hospital, Denmark, the Region of Southern Denmark, the Municipality of Odense, Denmark, the University of Southern Denmark, Odense Patient data Exploratory Network (OPEN), Denmark, the Danish Research Council (4004-00352B_FSS), Novo Nordisk Foundation, Denmark (grant no. NNF19OC0058266 and NNF17OC0029404), Sygeforsikring Danmark (journalnr. 2021-0173), the Collaborative Foundation between Odense University Hospital and Rigshospitalet, and Helsefonden. There is no conflict of interest of any author that could be perceived as prejudicing the impartiality of the research reported. N/A.

SIRT1 Regulates Mitochondrial Damage in N2a Cells Treated with the Prion Protein Fragment 106-126 via PGC-1α-TFAM-Mediated Mitochondrial Biogenesis.

Mitochondrial damage is an early and key marker of neuronal damage in prion diseases. As a process involved in mitochondrial quality control, mitochondrial biogenesis regulates mitochondrial homeostasis in neurons and promotes neuron health by increasing the number of effective mitochondria in the cytoplasm. Sirtuin 1 (SIRT1) is a NAD+-dependent deacetylase that regulates neuronal mitochondrial biogenesis and quality control in neurodegenerative diseases via deacetylation of a variety of substrates. In a cellular model of prion diseases, we found that both SIRT1 protein levels and deacetylase activity decreased, and SIRT1 overexpression and activation significantly ameliorated mitochondrial morphological damage and dysfunction caused by the neurotoxic peptide PrP106-126. Moreover, we found that mitochondrial biogenesis was impaired, and SIRT1 overexpression and activation alleviated PrP106-126-induced impairment of mitochondrial biogenesis in N2a cells. Further studies in PrP106-126-treated N2a cells revealed that SIRT1 regulates mitochondrial biogenesis through the PGC-1α-TFAM pathway. Finally, we showed that resveratrol resolved PrP106-126-induced mitochondrial dysfunction and cell apoptosis by promoting mitochondrial biogenesis through activation of the SIRT1-dependent PGC-1α/TFAM signaling pathway in N2a cells. Taken together, our findings further describe SIRT1 regulation of mitochondrial biogenesis and improve our understanding of mitochondria-related pathogenesis in prion diseases. Our findings support further investigation of SIRT1 as a potential target for therapeutic intervention of prion diseases.

Cardiac Autonomic Neuropathy Is Not Associated with Apolipoprotein E Gene Isoforms in the Kazakh Population: A Case-Control Study.

The absence of an early diagnosis of cardiac autonomic neuropathy might increase the risk of the disease, progressing to an irreversible stage. Therefore, this study aims to investigate the APOE gene isoforms in patients with cardiac autonomic neuropathy to identify early markers for predicting this disease in the Kazakh population. A total of 147 patients with cardiac neuropathy and 153 controls were examined in this case-control study. Patients were genotyped for two polymorphisms of the APOE gene using real-time PCR. Statistical calculations were performed using binary logistic regression. As a result of our study, we found that there was no statistically significant difference in the frequency of any APOE gene isoforms (APOE (ε2/ε2), APOE (ε2/ε3), APOE (ε2/ε4), APOE (ε3/ε3), or APOE (ε4/ε4)) between the patient group and the control group (p = 0.69, p = 0.64, p = 0.19, p = 0.22, p = 0.97, respectively). Thus, cardiac autonomic neuropathy is not associated with APOE gene isoforms in the Kazakh population.

Serum Interleukin-6 as an Early Indicator of Trauma Complications.

Trauma is a major global health issue, associated with high mortality and complications like inapparent hypoxia, fat embolism syndrome (FES), sepsis, and multiple organ dysfunction syndrome (MODS). Early identification of high-risk patients is crucial but challenging. Serum interleukin-6 (IL-6), a key inflammatory cytokine, has shown potential as a biomarker for predicting adverse outcomes in trauma. IL-6 levels typically increase rapidly following trauma, peaking within 12 to 24 hours. Despite its potential role, there is limited research on the effectiveness of IL-6 as an early marker for trauma-related complications. This study aims to assess whether monitoring serum IL-6 levels at specific intervals after trauma can aid in early risk assessment and predict the development of these complications. This prospective observational cohort study at North Eastern Indira Gandhi Regional Institute of Health and Medical Sciences (NEIGRIHMS) included 119 trauma patients aged 19-65 years, admitted within 12 hours of injury. Venous blood samples (5 mL each) were collected at 12 and 24 hours for IL-6 and C-reactive protein (CRP) analysis. Injury severity score (ISS) was assessed for all the patients upon arrival to the emergency department at NEIGRIHMS and was categorized as mild, moderate, severe, and very severe. Inapparent hypoxia, FES, sepsis, and MODS were assessed using pulse oximetry, Gurd's criteria, quick sequential organ failure assessment (qSOFA) score, and Marshall's multiple organ dysfunction score, respectively. Among the participants, 21.85% developed complications; primarily inapparent hypoxia. Serum IL-6 levels were significantly elevated in individuals with complications at both 12 hours (p < 0.001) and 24 hours (p < 0.001) post-trauma. At the 12-hour mark, serum IL-6 demonstrated a sensitivity of 92.3% and a specificity of 78.5%, with a cut-off value of 37.26 pg/mL. By 24 hours, the sensitivity increased to 96.2% and the specificity to 87.1%, with a cut-off value of 55.08 pg/mL. Patients with MODS had the highest IL-6 levels, with medians of 270.87 pg/mL at 12 hours and 826.10 pg/mL at 24 hours. A strong correlation was observed between serum IL-6 at 24 hours and the ISS(rs = 0.725, p < 0.001). At 12 hours, there was a moderate correlation between serum IL-6 and CRP (rs = 0.488, p < 0.001). By 24 hours, this correlation strengthened to a strong level (rs = 0.749, p < 0.001). The significant association of serum IL-6 levels with both ISSand CRP highlights its potential role in assessing trauma severity. The high sensitivity and specificity of IL-6 at the 24-hour make it a valuable biomarker for the early detection of trauma-related complications.

Cardiac Involvement in LAMA2-Related Muscular Dystrophy and SELENON-Related Congenital Myopathy: A Case Series.

LAMA2-related muscular dystrophy (LAMA2-MD) and SELENON-related myopathy (SELENON-RM) are two rare neuromuscular diseases characterized by proximal and axial muscle weakness, scoliosis, spinal rigidity, low bone quality and respiratory impairment. Cardiac involvement has previously been described in retrospective studies and case reports, but large case series and prospective studies in unselected cohorts are lacking. The objective of this study is to conduct prevalence estimations, perform cardiac phenotyping, and provide recommendations for clinical care. In this case series including two time points, we conducted comprehensive assessments with electrocardiography (ECG) and transthoracic echocardiography (TTE). ECGs were systematically assessed for a large subset of variables. TTE included left and right ventricular ejection fraction (LVEF/RVEF) and left ventricular global longitudinal strain (GLS), the latter being a more early and sensitive marker of left ventricular dysfunction. 21 LAMA2-MD (M = 5; 20±14 years) and 10 SELENON-RM patients (M = 7; 18±12 years) were included. In most patients, QRS fragmentation and Q waves, markers of heterogeneous ventricular activation, were present both at baseline and at follow-up. GLS was abnormal (age specific in children, > -18% in adults) in 33% of LAMA2-MD and 43% of SELENON-RM patients at baseline. Reduced LVEF (<52% in males, <54% in females and <55% in pediatric population) was observed in three LAMA2-MD patients at baseline and in none of the SELENON-RM patients. GLS and LVEF did not change between baseline and follow-up. RVEF was normal in all patients. ECG abnormalities and abnormal GLS are prevalent in LAMA2-MD and SELENON-RM, yet abnormal LVEF was only seen in LAMA2-MD patients. One LAMA2-MD patient had a clinically relevant deterioration in LVEF during 1.5-year follow-up. We advise routine screening of all patients with LAMA2-MD or SELENON-RM with ECG and echocardiography at diagnosis, minimally every two years from second decade of life and if new cardiac signs arise.