Better evaluation of the contribution of the main diseases, injuries, and risk factors for mortality and life expectancy is crucial for more efficient policy making at the national and subnational levels in Iran. The aim of this study is to assess the effect of emerging causes of mortality on health, specifically COVID-19, which can help policy makers implement preventive measures in similar situations. In this systematic analysis of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2023, we present estimates of cause-specific mortality at the national and subnational levels in Iran from 1990 to 2023. New to this iteration of GBD, we present a decomposition analysis of the contribution of specific causes of death to net gain or loss in life expectancy across 31 provinces of Iran. We used an array of data sources including censuses, vital registration, and surveys for national and subnational estimates. The two leading causes of death in Iran were ischaemic heart disease and stroke in both 1990 and 2019. However, in 2020 and 2021, the COVID-19 pandemic displaced the leading causes of death, ranking first with age-standardised mortality rates of 286·2 deaths (95% uncertainty interval 267·9-310·5) per 100 000 in 2020 and 250·0 deaths (233·2-272·5) per 100 000 in 2021. COVID-19 ranked second and tenth in 2022 and 2023, respectively. Life expectancy at birth for both sexes combined declined from 78·0 years (77·7-78·1) in 2019 to 74·3 years (74·0-74·4) in 2020. It steadily recovered to 78·8 years (78·5-79·2) in 2023. COVID-19 was the main cause of loss in life expectancy, by 4·19 years, between 2019 and 2020. There was a net gain of 12·4 years in life expectancy in Iran from 1990 to 2023. The net gain at the national level can be mostly attributed to reduced mortality from ischaemic heart disease (2·61 years), stroke (1·63 years), neonatal disorders (1·26 years), transport injuries (0·88 years), and neoplasms (0·64 years). The decline in mortality rates of major causes continued to 2023 despite the pandemic. An exception was Alzheimer's disease, which showed a 4·0% increase in rate between 2019 and 2023 and led to a net loss of 0·04 years in life expectancy since 1990. Diabetes led to a net loss of 0·09 years since 1990. There were variations between provinces in terms of age-standardised rates and the net change in life expectancy before and after the COVID-19 pandemic. The COVID-19 pandemic disrupted the rising trend of life expectancy in Iran, varying across provinces. Findings show that the health-care infrastructure and policies in Iran were not efficient in controlling the pandemic in 2020 and 2021, mainly due to inadequate vaccination coverage and timeliness, specifically for vulnerable subgroups. Sanctions may have aggravated the effect of COVID-19 on loss in life expectancy of Iranians. Despite the pandemic, the declining trend in age-standardised rates for top causes of mortality has continued to 2023, leading to a full recovery of life expectancy and underscoring the ultimate resilience of Iran's health system. Gates Foundation.
Sickle cell disease (SCD) is a severe, inherited hemoglobin disorder characterized by chronic hemolysis, vaso-occlusive crises (VOCs), and systemic inflammation. Hydroxyurea is the standard conventional pharmacotherapy for SCD, but it has certain limitations, necessitating the need to explore other safe and effective treatment options for SCD. Ayurveda interventions offer a potential therapeutic approach complementary to conventional medicine for SCD management, with anti-inflammatory, immunomodulatory, and hematopoietic properties. This randomized controlled trial will evaluate the efficacy and safety of an Ayurvedic therapeutic regimen as an adjunct to hydroxyurea in SCD management, assessing its impact on hematological parameters, inflammatory biomarkers, VOC frequency, and overall quality of life. A PROBE (Prospective, Randomized, Open-Label, Blinded End Point) study will be conducted on individuals of any gender aged 18 years or older and diagnosed with SCD (with hemoglobin S levels more than 60% and a history of at least 1 VOC per year over the past 3 y). Individuals with acute VOC or any severe infection requiring hospitalization, a history of significant comorbidities, or hematopoietic stem cell transplantation will not be considered. The study will be conducted at the All India Institute of Medical Sciences, Bhopal, India. A total of 100 participants will undergo random assignment in a 1:1 ratio to receive either an Ayurveda regimen (Dadimadi Ghrita, Punarnavadi Mandura, and Vasaguduchyadi Kwatha) as an add-on to hydroxyurea or hydroxyurea alone for 8 months. The primary outcome will be a change in hemoglobin electrophoresis parameters (hemoglobin S, fetal hemoglobin, and adult hemoglobin) and the frequency of VOC episodes over 8 months. The secondary outcome measures include changes in the levels of proinflammatory markers (interleukin-6, interleukin-8, C-reactive protein, and transforming growth factor-β) and lactate dehydrogenase, frequency of hospitalization for VOCs and blood transfusions, and health-related quality of life (Short Form-8 Health Survey questionnaire). Safety will be evaluated by recording the incidence of adverse events and changes in liver and kidney function tests from baseline. The recruitment of study participants was initiated on November 1, 2023. By the second week of February 2025, 83 participants had been enrolled in the study. The final study is expected to be complete by December 31, 2025. We will start the analysis of the study outcomes in February 2026, and the publication of the final results is expected by August 2026. This randomized controlled trial protocol outlines a rigorous study design aimed to explore the potential benefits of an integrated therapeutic regimen comprising Ayurveda interventions and standard conventional care in the long-term management of SCD through validated clinical and laboratory parameters. The outcomes of this study can address the needs and challenges associated with SCD management and inform future management protocols.
The widely used chemotherapeutic drug cisplatin (CDDP) is an integral part of the preoperative chemotherapy protocol for high-grade osteosarcoma (OS). However, despite an aggressive treatment regimen, drug refractoriness is a major hindrance to successful therapy. We previously identified key transcriptomic alterations that support the survival of OS cells following CDDP exposure. In the present study, our in vitro analyses further demonstrate that CDDP treatment promotes an adaptive, ROS-dependent enrichment of the repressive histone mark H3K27me3 at the upstream promoter regions of growth-associated genes such as CCNA2, as well as at the promoter of LATS1, a negative regulator of Yes-Associated Protein (YAP). This enrichment contributes to the transcriptional repression of these genes and is associated with growth arrest; notably, quenching of reactive oxygen species with N-acetyl cysteine (NAC) reversed this effect. Furthermore, reduced LATS1 expression was associated with increased nuclear localization of YAP. Importantly, pharmacological inhibition or genetic ablation of YAP attenuated the CDDP-induced accumulation of repressive marks. Mechanistically, YAP was found to colocalize and coimmunoprecipitate with EZH2, the catalytic member of the Polycomb Repressive Complex 2 (PRC2), suggesting a potential role for YAP in facilitating EZH2-mediated transcriptional repression. Consistent with these observations, inhibition of YAP or pharmacological reversal of the repressive chromatin state using a histone deacetylase (HDAC) inhibitor enhanced the sensitivity of the OS cells to low-dose CDDP treatment. Overall, the present study demonstrates an interplay among oxidative stress, epigenetics, and YAP in modulating OS cell fate post-CDDP exposure.
The root of the tree of life remains a controversial issue in the study of the origins and evolution of early life on Earth, and so is the origin and evolution of early biochemistry. Here, we describe the methodological procedures of reconstructing global phylogenies of early life based on RNA secondary structure, with the rooting of phylogenies developed and improved to ultimately gain insights into the deep evolution of ancient molecules. As an example, we explore the evolution of the transfer RNA (tRNA) molecules using statistical characters describing their structure. An evolutionary model of molecular accretion of tRNA modeled at the atomic level illustrates the experimental exercise.
Routine inpatient biochemical testing is often repeated without clinical indication. This quality improvement project aimed to reduce low-value biochemical testing in a Danish cardiology ward, targeting a 5% reduction within 6 months and sustainment for 12 months, without adverse changes in length of stay, discharges after noon, or 30-day readmissions. In a quality improvement project using the Model for Improvement and conducted through Plan-Do-Study-Act cycles, a multidisciplinary team implemented the following interventions: (i)targeted education for physicians and nurses e.g. clarifying of ordering responsibility; (ii)refinement of default biochemical profiles by removing non-essential tests (e.g. leukocyte count with differential, urea, hemoglobin) and curbing premature orders placed before the minimum re-testing interval; and (III)a centrally placed feedback dashboard. The primary outcome was the number of biochemical tests, adjusted for hours of hospitalization and caseload. Secondary outcomes were the proportion of results outside reference intervals and re‑testing intervals for common biochemical tests. Safety balancing measures were length of stay, discharges after noon, and 30-day readmissions. The pre-intervention period ran from February 27th, 2022, to February 26th, 2023, post-intervention period from March 5th, 2023, to February 2nd, 2025. After biochemical profile refinement in June 2023, median number of biochemical tests decreased 54% (hours of hospitalization adjusted) and 48% (caseload adjusted). The proportion of test results outside reference ranges increased 21%. Re-testing intervals lengthened for hemoglobin and troponin-I. Balancing measures showed no adverse signals, and effects persisted into February 2025. A pragmatic, simple bundle halved routine biochemical tests and increased diagnostic yield without measurable harm.
Brazil is the second highest worldwide regarding new cases of Hansen's disease (HD), with over 20,000 diagnoses annually. Late detection often results in severe disabilities that impair daily functioning, reduce work capacity, and increase reliance on state financial assistance, which contribute to public expenditure. This study quantified the economic impact of Hansen's disease by analyzing social security benefits in relation to the disease's epidemiological profile from the late 20th century to the first two decades of the 21st century. This retrospective, longitudinal, descriptive study applied a cost-of-illness (COI) approach from a societal perspective. Indirect costs attributable to Hansen's disease were estimated using social security transfer payments as proxies for productivity loss. Administrative data from the National Institute of Social Security were obtained for all benefits granted between 2000 and 2019. These records included temporary, long-term, and lifetime benefits, which were categorized by type, duration, sex, geographic location, and total financial disbursement. The economic evaluation considered both annual and accumulated payments over the study period. To contextualize the burden of disease, benefit data were analyzed alongside epidemiological indicators from the Ministry of Health's Notifiable Diseases Information System, allowing the examination of the relationship between Hansen's disease trends and variations in benefit concessions. Systematic variations were identified in the number of benefits granted, with a marked increase until 2005, followed by an abrupt drop and a subsequent gradual decline. These fluctuations were influenced by exogenous monetary and political factors and changes in benefit requests. Long-term benefits followed the same pattern, stabilizing at approximately 1,000 grants per year. Temporary benefits showed recurrent oscillations and later declined from 4,283 in 2000 to 3,525 in 2019, remaining substantially more frequent than lifetime benefits. Data segmentation showed the following: (I) age at entry into the social security system peaked at 45 to 55 years for both sexes; (II) men received 2.7 times more benefits; (III) benefit duration was similar for men and women, with 52.0% ending within 18 months and 85.0% within 120 months; (IV) the Southeast region accounted for 21.8% of benefits and 24.8% of new cases; and (V) total payments increased over 60-fold, reaching US$ 91 million in 2019. Social security expenditures for beneficiaries with Hansen's disease totaled US$ 924.6 million, representing a substantial economic burden relative to the disease's epidemiological characteristics in Brazil. Although incidence has declined and benefit concessions have gradually decreased, partly due to improvements in administrative processes and Hansen's disease control, the financial impact remains significant. Given that Hansen's disease is curable, and treatment is freely available in the public health system, current resource allocation prioritizes compensation for productivity loss over early diagnosis and prevention. The identified disparities, including gender differences and concentration of benefits in wealthier regions, warrant further investigation to inform mitigation strategies.
The deep entanglement of biomolecular structure and function with aqueous systems supports the view that water actively sculpted both molecules and processes during the origins of life and continues to constrain evolution today. Nature's rules of biochemistry and biophysics have survived for nearly 4 billion years. The in vivo roles of water, the architectures of biopolymer backbones and side chains, the structure and function of ribose, ATP, the translation system, the genetic code, and certain inorganic cations persist unchanged. Hydrogen bonding and the hydrophobic effect predate life on Earth altogether. Here we examine fundamental forces that established, shaped, and continue to constrain biochemistry and biophysics. The results support a model in which life emerged through water-based selection among diverse molecules and molecular ensembles, with molecular fitness defined by behaviors in and interactions with water. Life is thus composed of molecules that cooperate with, resist, and exploit the unique properties of water. Biological molecules employ chemical strategies that enable selective and controlled persistence in aqueous environments, a phenomenon we classify as recalcitrance. Molecular assembly reduces conformational heterogeneity, constrains dynamics, and sterically excludes reactive agents, including water and hydrolytic enzymes. By this mechanism, lifetimes of folded proteins, structured RNAs, assembled phospholipids, and polysaccharides are mediated by their organizational states.
Streptococcus suis is a serious zoonotic pathogen responsible for rapid progression and deadly infections in both humans and pigs. With an increasing number of reported cases and considering the limitations of standard routine identification, a simple, rapid, and cost-effective approach is needed. In this study, a label-free colorimetric assay based on gold nanoparticles (AuNPs) was applied with a specific aptamer, R8-su12. This assay offered simplified detection through observable color change, enabling visual analysis by the naked eye or assessment via UV-Vis spectrophotometry. Under the optimal assay conditions, the detection procedure was carried out within 45 min. The reaction of the aptasensor and other bacterial species, including Staphylococcus aureus, S. pneumoniae, S. pyogenes, Pseudomonas aeruginosa, Escherichia coli, Enterococcus faecium, and E. faecalis, was not present, indicating the specificity of this assay. Moreover, the aptasensor exhibited high sensitivity with a limit of detection (LOD) at 1 CFU of S. suis and had broad reactivity with S. suis serotypes 1, 1/2, 9, and 14, as well as with S. suis isolated from clinical specimens. Thus, this aptasensor demonstrates proof-of-concept feasibility including clinical sample testing before practical implementation. It holds promise as a practical tool for the early screening and outbreak management of S. suis in a variety of settings, such as clinical laboratories, food safety, and the environment.
A dual-mode headspace and liquid GC-MS approach was employed to analyse the essential oil extract of Cinnamomum camphora variant grown in Taiwan. Two β-cyclodextrin GC columns (Cydex-B and Rt-βDEXsm™) were compared for chiral separation. A note that Rt-βDEXsm™ column showed superior resolution for borneol and camphor isomers, and reveals that (+) borneol (31.7%), eucalyptol (11.4%), and (+) camphor (8.4%) as the major constituents in the essential oil. Distinct chemical profiles emerged from each sampling mode, headspace detects light aroma active volatiles like (+)-α-pinene and (-)-camphene, whereas liquid sampling reveals heavier oxygenated compounds (borneol and camphor) and 13 additional analytes. Such differences in volatile profiles are particularly important for aromatherapy and inhalation-based applications. Analysis of commercial (+)-borneol revealed the presence of (-)-isoborneol, highlights the importance of careful assessment of reference materials in chiral analysis. The direct analysis of Cinnamomum camphora variant grown in Taiwan showed high percentage of (+)-borneol, which suggests that locally grown variant can serve as a dependable source of natural (+)-borneol to minimize imported raw materials and maintain high quality standards.
Chitosan (CS) is a cationic linear polysaccharide rich in functional groups (-OH and -NH2) that enable diverse biochemical interactions, making it a promising candidate for the electrochemical detection of dopamine (DA), uric acid (UA), and ascorbic acid (AA). However, its poor electrical conductivity and limited thermal stability restrict its standalone performance, thus requiring the formation of composites with other materials. In this study, we report the synthesis of a chitosan/Fe3O4/graphene nanoplatelet (CS@Fe3O4/GNP) nanocomposite for the highly sensitive and selective electrochemical detection of DA, UA, and AA. The nanocomposite integrates the superior conductivity and high surface area of GNP, the electrocatalytic activity of Fe3O4 nanoparticles, and the biocompatibility and adhesive nature of chitosan. The CS@Fe3O4/GNP composite with various CS concentration (0.0625, 0.125, and 0.25%) was synthesized via a facile in-situ co-precipitation method. Electrochemical studies demonstrated that the 0.25% CS@Fe3O4/GNP-modified glassy carbon electrode (GCE) exhibited excellent detection performance toward DA, UA, and AA, with limits of detection (LOD) of 28.37 nM (range 30-488 nM), 566.27 nM (range 0.98-15.6 μM), and 26.54 μM (range 31.25-500 μM), respectively. The sensor also achieved recovery rates of 87.83-92.68% (DA), 82.41-91.61% (UA), and 88.83-95.04% (AA) in human blood serum samples.
Breast cancer remains the most prevalent malignancy among women worldwide, contributing significantly to global cancer-related morbidity and mortality. It is responsible for nearly 15% of all female cancer fatalities. It has a higher prevalence in developed nations. The present study aimed to assess the predictive utility of Atherogenic index of plasma (AIP) in carcinoma breast cases in correlation with oxidative stress, immuno-biomarkers and body mass index (BMI). Prospective interventional study. The Prospective Interventional Study was conducted in the Department of Biochemistry, Jawaharlal Nehru Medical College and the Histopathology and Radiology division of the Department of Pathology, JNMC, in collaboration with the Department of Surgery, JNMC and Acharya Vinoba Bhave Rural Hospital, Wardha, for the duration of 3 years. Total 81 samples were recruited for the study. The patient's history was taken, and a proper medical examination was carried out. The SPSS program for Windows, version 26.0 (SPSS, Chicago, Illinois, USA), was employed to conduct the statistical analysis. Categorical variables were analysed using either the Chi-square test or Fisher's exact test, while continuous variables were compared using the unpaired t -testing method. P ≤0.05 was deemed to be statistically significant. A significant positive correlations between AIP and various parameters, including BMI ( r = 0.591, P < 0.001), superoxide dismutase ( r = 0.407, P < 0.001), high-sensitivity C-reactive protein ( r = 0.507, P < 0.001), Interleukin-10 (IL-10) ( r = 0.706, P < 0.001) and IL-18 ( r = 0.652, P < 0.001). Furthermore, a strong positive correlation with lipid profile was observed. AIP could serve as a useful surrogate marker to assess not only lipid imbalance but also broader systemic changes that accompany breast cancer.
Early detection of cognitive decline may be effective in reducing the adverse impacts of Alzheimer's disease and related dementias (ADRD). Given that functional declines precede ADRD evaluation and diagnosis, regular assessments of everyday function are an avenue for detecting cognitive performance changes. While app-based measures of everyday function and cognition are promising tools for early detection, perceptions of these tools' value remain unexamined. This study explored perceptions of an app-based measure of everyday function (i.e., comfort with sharing performance data and perceived utility in healthcare) with community-dwelling midlife and older adults in South Carolina, United States (N = 131, Mage = 67.08 years). Participants completed daily tasks through a mobile app objectively measuring everyday function then shared their feedback through a semi-structured interview. Our thematic analysis found that interest and confidence in utilizing this technology was connected to beliefs around the value of having real-time information about one's cognitive performance, experiences with healthcare providers, and trust in technology security and accuracy. Additionally, some adults have not thought critically about the role of these technologies in their healthcare. As health-tracking technology expands in cognitive healthcare, researchers and practitioners must be aware of midlife and older adults' perceptions and educate users on its potential function.
Brivaracetam belongs to antiepileptic drugs and due to the interindividual variability, therapeutic drug monitoring is recommended, especially when used in polytherapy and in patients with altered pharmacokinetics. The aim of this study is to develop and validate a high-performance liquid chromatography (HPLC) method with a diode array detector (HPLC-DAD) that is suitable for therapeutic monitoring of brivaracetam and assess method's greenness. High-performance liquid chromatography method was validated according to the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) and Clinical and Laboratory Standards Institute (CLSI) guidelines. Preparation of serum samples was performed by liquid-liquid extraction using ethyl acetate:hexane (1:1) with chloramphenicol as internal standard. Separation was achieved using a 5 µm 4.6 x 250 mm C18 Shim-pack GIST column with the mobile phase consisting of a 50 mM phosphate buffer (pH = 4.5) and organic mixture of acetonitrile and methanol (8/3; v/v). Brivaracetam is quantified at 210 nm. AGREEprep was utilized for the method's greenness scoring. The developed HPLC-DAD method is linear over a concentration range of 0.25-20.00 µmol/L. Repeatability was lower than 7.20%, intermediate precision was lower than 8.30%, and within-laboratory precision was lower than 10.50%, except for the values at the lower limit of quantification (0.25 µmol/L) which were still below 20.00%. Bias was lower than ± 10.60%. The method displays very high specificity at 210 nm. The result for method greenness was 0.4. The validation parameters were within the acceptance criteria of the ICH and CLSI guidelines. The developed method is simple, selective, reproducible, more sensitive than other published HPLC methods and suitable for the therapeutic drug monitoring of brivaracetam.
Carbohydrate Response Element Binding Protein (ChREBP) is a transcription factor known to regulate glucose metabolism and other metabolic processes in various tissues, but its role in lung adenocarcinoma (LUAD) remains poorly understood. In this study, we investigated ChREBP expression and its role in regulating gene expression in LUAD cell lines. Using RT-qPCR, we assessed the expression of ChREBP-α and ChREBP-β isoforms in NCI-H1975, NCI-H1650, and NCI-H2228 LUAD cell lines. The NCI-H1975 cells exhibited the highest levels of both ChREBP isoforms, with a particularly pronounced expression of ChREBP-β. To explore the regulatory role of ChREBP, we generated NCI-H1975 cells with inducible expression of a dominant-negative mutant of human ChREBP (dnChREBP). Overexpression of dnChREBP led to a significant reduction in colony formation and impaired cell migration. Transcriptome analysis revealed 57 upregulated genes and 593 downregulated genes in dnChREBP-expressing cells compared to control cells. Functional annotation and gene set enrichment analysis revealed that the enriched genes were associated with cancer-related processes, including cell proliferation and epithelial-to-mesenchymal transition (EMT). Gene network analysis highlighted 17 downregulated hub genes, with 8 of these genes being associated with EMT. Interestingly, ChREBP and its transcriptionally regulated genes, including 4 top downregulated genes, 5 top upregulated genes, and 5 hub genes identified in NCI-H1975 cells overexpressing dnChREBP, showed significant prognostic value, as their expression levels correlated with overall survival in LUAD patients. Our findings suggest that ChREBP regulates distinct transcriptional programs in LUAD cells and ChREBP and its regulatory network may play a potential role in LUAD progression and patient outcomes.
Cardiac troponin I (cTnI) is a cardiac specific biomarker of myocardial damage in humans, dogs, and cats. The ADVIA Centaur XP High-Sensitivity Troponin I assay (AC-cTnI-HS) has been validated for use in humans and dogs, but not for use in cats. The study objective was to analytically validate the AC-cTnI-HS assay for use in cats and to evaluate cTnI measurements in healthy cats compared to those with cardiac disease to assess the clinical utility of this assay. Surplus serum samples from cats were used for analytical validation. Intra- and inter-assay variability, dilutional parallelism, and spiking recovery were assessed. Serum samples from 106 client-owned cats were evaluated. This group was comprised of 51 clinically healthy cats (structurally normal echocardiogram, normal systemic blood pressure, and unremarkable complete blood count and biochemistry profile), 25 cats with stage B1 hypertrophic cardiomyopathy, 7 with stage B2 hypertrophic cardiomyopathy, 7 with stage C cardiomyopathy of any type, 8 with congenital heart disease, and 8 cats with transient myocardial thickening and/or suspected to have myocarditis. Inter-assay and intra-assay coefficients of variation were between 2.7-8.3% and 1.5-4.0%, respectively. The mean ± standard deviation observed to expected ratios for dilutional parallelism and spiking recovery were 124.3 ± 42.8% and 92.9 ± 6.2%, respectively. Healthy cats had significantly lower cTnI concentrations than cats with hypertrophic cardiomyopathy stage B1 (P = 0.012), stage B2 (P = 0.004), or any cardiomyopathy ACVIM stage C (P = 0.002). The AC-cTnI-HS assay is precise, reproducible, linear, and accurate for measurement of cTnI concentrations in serum from cats. This study confirms that measurement of serum cTnI holds promise to have clinical utility as it was able to detect differences in serum cTnI concentrations between healthy cats and those with cardiac disease.
Measurement of plasma melatonin is important in studies of sleep disorders and neurodegenerative diseases and is also increasingly associated with cardiometabolic diseases and cancer. There is currently an increase in the usage of melatonin in the population. However, commercially available and reliable melatonin assays are lacking. Here we present a rapid and simple LC-MS/MS-based method for plasma melatonin in a clinical laboratory. The method demonstrates a limit of detection of 5 ng/L and inter-assay imprecision < 16%. This newly developed LC-MS/MS method was applied to 216 plasma samples collected from 24 men as part of The Bispebjerg study of diurnal variation. The results showed increased melatonin concentrations during the nighttime and fitted well to a diurnal rhythm curve (p < 0.0001). The LC-MS/MS assay correlated significantly with commercial RIA (p < 0.03) but demonstrated a substantial bias. We conclude that our simple LC-MS/MS method can detect biologically relevant changes in plasma melatonin and may enable insight into circadian biology in humans. We propose that the observed differences between melatonin assays could be clarified by the distribution of reference plasma melatonin material and that additional studies are needed to fully understand the minimum circulating levels of melatonin in the daytime.
Although next-generation RNA sequencing (RNA-seq) is increasingly incorporated into germline cancer predisposition testing, its diagnostic utility is often limited by low expression of many clinically relevant genes. To improve RNA yield and transcript representation for targeted RNA-seq, we optimized a simple protocol based on short-term lymphocyte culture prepared directly from whole blood collected in Li-heparin tubes. We systematically evaluated biological reproducibility and pre-analytical sample handling variability and demonstrated that whole blood can be stored at 4 °C for up to 5 days prior to lymphocyte cultivation without compromising RNA quality/gene expression. Gene expression was comparable for RNA isolated from K2EDTA and Tempus tubes, whereas short-term lymphocyte culture resulted in a substantial increase in expression of clinically important genes including BRCA1, BRCA2, RAD51C, RAD51D, PALB2, CHEK2, and multiple Fanconi anaemia genes otherwise low expressed in whole blood. Cultivation for 3-5 days did not significantly affect lymphocyte gene expression, providing flexibility for routine dia-gnostics. The protocol also enables inhibition of nonsense-mediated decay to facilitate analysis of variants causing premature termination. As a proof of principle, we characterized the splicing impact of FANCA c.2602-3C>G variant (located in intron 27) using cultured lymphocytes from its carrier. The variant causes deletion of six nucleotides in the mature transcript (ΔE28p(-6)/r.2602_2607del), resulting in an in-frame deletion (p. Gln869_Phe870del). This spliceogenic effect was reliably detectable only in cultured lymphocytes preferentially expressing the full-length FANCA transcript. Overall, short-term lymphocyte culture re-presents a simple and flexible RNA source that enhances variant interpretation in clinical RNA-seq analyses.
The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has had immense global consequences, leading to widespread illness, deaths, and devastated economies. Despite this, Africa has experienced a high prevalence of asymptomatic coronavirus disease 2019 (COVID-19) and mild cases. While reported cases and deaths have been lower, limited testing and undiagnosed infections make it difficult to determine the true burden of the disease. Understanding the unique immune response and the variations in genetics affect COVID-19 outcomes in African populations is important for shaping future public health responses. This review examines key immune factors and genetic variations in key host proteins that may help explain why COVID-19 was less severe in Africa. A systematic review was conducted following PRISMA guidelines to identify studies published between 2019 and January 2026 that investigated immunological responses and genetic variations associated with COVID-19 in African populations. Literature searches were performed in PubMed, Scopus, and African Journals Online (AJOL). Inclusion criteria focused on studies reporting responses from cytokines, T-cells, antibodies or host genetic factors. After screening 4,170 records and removing duplicates, 420 studies were assessed for abstracts, and 240 full texts were reviewed. A total of 40 studies were included, and data synthesized narratively due to heterogeneity in study designs and outcomes. Of the 40 studies analyzed from 19 African populations, 26 focused on immunological responses and 9 on host genetic factors. Immune studies revealed widespread pre-existing immunity, including cross-reactive antibodies (especially to the N proteins) and polyfunctional T-cell responses, likely shaped by exposure to malaria, helminths, and other coronaviruses. Severe COVID-19 cases showed elevated IL-6, TNF-α, and IFN-γ, while asymptomatic individuals had broader, milder cytokine profiles. Antibody responses were robust across disease severities, with long-lasting IgG activity. Genetic studies identified HLA-B41, B42, C16, and C17 as risk alleles, while HLA-DQB106, DQB103, and B*15 conferred protection. ACE2 polymorphisms including rs2285666, rs73635825 were reportedly prevalent in Africans and were linked to varied ACE2 expression, viral load, and disease severity. The findings suggest that immune and genetic adaptations in African populations may have modulated susceptibility and severity of SARS-CoV-2 infection outcomes in Africans. https://www.crd.york.ac.uk/PROSPERO/view, identifier CRD420251121731.
Primary malignant brain tumors (PBT) impose substantial burdens on patients and caregivers. Caregivers are essential in the delivery of outpatient care for patients with PBT but experience high levels of fatigue, distress, and health decline. Although exercise is known to improve outcomes in cancer patients, interventions tailored specifically to the PBT patient-caregiver dyad remain limited. Dyadic intervention, as well as exercise oncology, are emerging areas of active research in neuro-oncology. This scoping review incorporates both principles to evaluate the existing literature on exercise interventions on primary brain tumor patient-caregiver dyads. We conducted a comprehensive search of MEDLINE (PubMed), Embase, CINAHL (EBSCO), Rehabilitation & Sports Medicine (EBSCO), and Cochrane Central (Ovid) in December 2025 for studies involving exercise interventions that included adult PBT patients and caregivers. Of the 1126 records screened, eight studies were included: four yoga-based interventions (three feasibility trials and one ongoing multicenter RCT), one pilot ski-based intervention, and three aerobic and resistance training-based interventions (two qualitative and one ongoing trial). The interventions were safe and feasible, with high adherence and retention. The preliminary reported benefits included improvements in fatigue, sleep, quality of life, and caregiver distress for the dyads. Videoconference delivery was effective, particularly during the COVID-19 pandemic. The eight included studies comprised 5-67 dyads, with four being single-arm feasibility studies. Current literature on dyadic exercise intervention in neuro-oncology consists primarily of small-scale feasibility and pilot studies. Initial findings have demonstrated that such interventions are safe. However, preliminary efficacy remains limited due to the risk of bias and lack of statistical power. Larger randomized clinical trials with objective endpoints are needed to define efficacy and guide evidence-based protocols.
Early recognition of pediatric sepsis remains challenging due to the limited diagnostic performance of conventional biomarkers. This study aimed to evaluate the kinetic profiles of pentraxin-3 (PTX3) and presepsin and their associations with the Phoenix Sepsis Score (PSS) and hemodynamic support requirements. In this prospective single-center study, 40 children with suspected sepsis and 66 healthy controls were enrolled. Patients were classified according to the Phoenix Sepsis Score (PSS) as infection (PSS < 2), sepsis (PSS ≥ 2), and septic shock. Serum PTX3, presepsin, C-reactive protein (CRP), and procalcitonin (PCT) levels were measured at admission and at 48 h, and their temporal patterns and clinical correlations were analyzed. Baseline PTX3 levels were significantly higher in septic children than controls (p < 0.001), whereas presepsin showed no difference (p = 0.513). PTX3 varied across clinical severity groups (p < 0.001), while CRP and PCT did not. PTX3 showed high diagnostic accuracy (AUC: 0.872; 95% CI: 0.787-0.958), whereas presepsin performed poorly (AUC: 0.578). Admission PTX3 correlated with vasopressor requirement (r = 0.548, p = 0.042). At 48 h, PTX3 decreased significantly (p < 0.001), while presepsin increased (p = 0.034).Conclusion: Our results suggest that PTX3 may provide added diagnostic utility compared with presepsin for early detection and severity assessment of pediatric sepsis. Its pronounced variation across clinical categories and strong correlation with hemodynamic support indicate that PTX3 could serve as a dynamic biomarker of vascular endothelial activation and subsequent stabilization. What is Known: • PTX3 and presepsin have been proposed as emerging biomarkers for sepsis, however evidence in pediatric populations remains limited • Existing pediatric studies report heterogeneous diagnostic performance for presepsin, and data on early kinetic behavior of PTX3 are scarce What is New: • This study demonstrates distinct temporal biomarker dynamics: PTX3 levels decline rapidly with clinical stabilization, whereas presepsin shows a significant late-phase increase within 48 h • When validated against the Phoenix Sepsis Score, PTX3 exhibits higher diagnostic reliability and a stronger correlation with vasopressor requirements than presepsin, suggesting that it may serve as a more specific indicator of vascular endothelial stress.