Vital Marker Research Articles

Fluoro-Fucosylation Enables the Interrogation of the Lea - LecB Interaction by BioNMR Spectroscopy.

Fucosylation patterns in cell-surface glycans are essential mediators of recognition and signalling. Aberrations in these signatures serve as vital diagnostic markers of disease progression, and so understanding fucose-protein interactions at the molecular level is crucial. Molecular editing of l-fucose (Fuc) at C2 with fluorine provides a platform to reconcile the ubiquity of fucosylation with the paucity of strategies to interrogate site-specific interactions. Through judicious introduction of a pseudo-equatorial fluorine [C(sp3)-F] adjacent to the anomeric position, β-selective fucosylation can be achieved with a range of diverse acceptors (>50:1): the selectivity of this process can be inverted through changes in the donor scaffold. Reaction development was driven by the desire to construct a fluorinated analogue of Lewis antigen a (F-Lea), in which fluorine replaces a key OH group at C2. Lea is a ligand for Lectin B (LecB) in the pathogen Pseudomonas aeruginosa and thus delineating the importance of key interactions in this complex has ramifications for drug discovery. Independent syntheses of Lea and F-Lea, and systematic bioNMR analyses with both glycans has unequivocally established the essential role of O2 of fucose in the Lea-LecB complex.

Improving prognostic evaluations in patients with stage IIIb light chain cardiac amyloidosis: role of haemodynamic parameters

BackgroundThere is no unified prognostic scoring system for light chain cardiac amyloidosis (AL-CA), particularly stage IIIb AL-CA. This study aimed to use invasive haemodynamic information to investigate markers that can more accurately evaluate the prognosis of patients with stage IIIb AL-CA.MethodsIn this retrospective cohort study, we conducted invasive haemodynamic measurements concurrently with myocardial biopsies to diagnose AL-CA. We used Cox regression analysis and time-dependent receiver operating characteristic curve analysis to study the associations between these measurements and overall mortality. Echocardiographic parameters were also recorded and analysed via logistic regression to explore their relationships with haemodynamic changes.ResultsAlthough traditional haemodynamic parameters, such as the cardiac index (CI), pulmonary artery wedge pressure (PAWP), pulmonary artery pressure, and vascular resistance, did not correlate with mortality, the PAWP/CI ratio emerged as a vital prognostic marker. Patients with a PAWP/CI ratio above 11 mmHg/L/min/m2 had markedly poorer survival. Kaplan‒Meier analysis highlighted the prognostic significance of the ratio, revealing distinct survival differences. Furthermore, logistic regression confirmed that echocardiographically measured pulmonary artery systolic pressure independently correlated with increases in the PAWP/CI ratio.ConclusionsIn stage IIIb AL-CA patients, the PAWP/CI ratio, which surpasses traditional haemodynamic indicators, significantly predicts all-cause mortality, emphasizing its prognostic value. Our findings suggest that echocardiography-derived PASP could alternatively reflect the PAWP/CI ratio.

Safety and efficacy of oral edible bird’s nest supplementation: Anti-inflammatory and immunomodulatory benefits for Arabian race stallions during exercise

Background and Aim: Exercise-induced oxidative stress and inflammation adversely affect the health and performance of athletic horses. This study aimed to evaluate the safety of edible bird’s nest (EBN) supplementation and its potential anti-inflammatory and immunomodulatory effects in Arabian race stallions during exercise. Key objectives included assessing EBN’s impact on hepatic and renal function, hematological profiles, and sialic acid (SA) levels. Materials and Methods: Two experiments were conducted on 18 Arabian stallions. In Experiment 1, six healthy stallions were divided into control (n = 2) and EBN-supplemented groups (n = 4) to assess safety. The treatment group received 10 g of EBN daily for 12 days. Vital signs, hematological parameters, and organ function markers were monitored. In Experiment 2, 12 stallions were divided into three groups: EBN (n = 3), Premier E® (n = 3), and control (n = 6). The exercise regimen included a daily 10-min walk, 10-min canter (30 km/h), and 10-min walk for 30 days. Blood samples were analyzed for hematological indices and SA levels pre- and post-exercise. Results: In Experiment 1, EBN supplementation demonstrated no adverse effects, maintaining normal hematological and vital parameters. Kidney and liver function tests revealed significantly reduced creatine kinase, total bilirubin, and aspartate aminotransferase levels in the EBN group. Experiment 2 showed higher SA levels post-exercise in the EBN group (p < 0.05) compared to Premier E® and control groups, suggesting enhanced anti-inflammatory response. Hematological indices, including mean corpuscular volume, red cell distribution width, and platelet counts, were significantly improved in the EBN group, indicating potential immunomodulatory benefits. Conclusion: EBN supplementation is safe and offers anti-inflammatory and immunomodulatory effects in Arabian race stallions, reducing exercise-induced hepatic and muscular stress markers while enhancing recovery. These findings support EBN as a valuable dietary supplement for performance horses. Further studies should explore long-term effects and molecular mechanisms to optimize its use in equine sports. Keywords: anti-inflammatory, arabian race stallions, edible bird’s nest, exercise stress, immunomodulatory.

Evaluation of Glutathione-Related Antioxidant Enzyme Activity in Patients with Polycystic Ovary Syndrome (PCOS) and Investigation of Clinical Correlations

The aim of this study was to illustrate the importance of glutathione-associated enzymes as vital markers of antioxidant/oxidant activity in PCOS disease. Our study comprised a cohort of forty patients, all of whom were above the age of 18 and had been diagnosed with Polycystic Ovary Syndrome (PCOS) according to the 2003 Rotterdam diagnostic criteria. In addition, we recruited forty healthy individuals who were free from any metabolic disorders. The Rotterdam diagnostic criteria for polycystic ovarian syndrome consist of three main factors: oligo-anovulation, clinical and/or biochemical hyperandrogenism, and ultrasonographic confirmation of polycystic ovaries. Individuals who satisfied at least two of the three criteria were categorized as having Polycystic Ovary Syndrome (PCOS). The levels of gamma-glutamyl transferase (GGT) and deltaglutamyl cysteine synthetase (ꝩ-GCS) in serum samples from both patients and controls were measured using the enzyme-linked immunosorbent assay (ELISA) technique. The study examined the levels of essential enzymes involved in glutathione metabolism in patients with PCOS. The enzymes displayed significant differences between PCOS patients and control groups, indicating that glutathione metabolism plays a pivotal role in the progression of this disease. After conducting correlation analysis, it is clear that enzymes have a reciprocal impact on the progression of the disease. The analytical findings suggest that there was no statistically significant disparity in GGT levels between the PCOS group and the control group (p>0.05). The levels of ꝩ-GCS were significantly higher in the PCOS group compared to the control group (P< 0.05). Considering the involvement of ꝩ-GCS in the process of glutathione catalysis, it is postulated that its concentrations would increase in response to an elevation in glutathione levels. Our analysis of the available evidence leads us to conclude that glutathione will exert a substantial influence on the mechanism of PCOS. Polycystic ovarian syndrome (PCOS) is a notable medical illness that frequently affects women and has a substantial influence on their overall well-being.

Ethnolinguistic Vitality in Minority Schoolscape

School is often said to be a representation of society because its primary aim is to promote integration into society. This study of the landscape elements of minority language schools suggests that this type of linguistic landscape may not only reflect a change in linguistic dominance but can also play a complementary role compared to the external, out-of-school world. In this paper, the authors attempt to explore a new way of interpreting the notion of the linguistic landscape of schools. The novelty of this approach lies in the application of a spatial theoretical concept, where the schoolscape is defined as an element of autopoietic space—a self-constituting spatial element. Within this autopoietic framework, markers of ethnolinguistic vitality in the linguistic landscape of minority language schools are identified, and a set of criteria is established that can be applied to other communities. The conclusions are drawn from a comparative analysis of the linguistic landscapes of Hungarian-medium schools in Romania, Slovakia, and Slovenia.

Added value of inflammatory markers to vital signs for predicting mortality in patients with suspected infection: external validation and model development.

It is crucial to identify high-risk patients with infectious conditions for appropriate management. We previously found that inflammatory markers added value to vital signs for predicting mortality in patients with suspected infection. In this study, the aim was to externally validate the added value of the inflammatory markers and to develop a new prediction model. For the external validation, consecutive adult patients with suspected infection admitted to the department of general medicine at two acute care hospitals were evaluated. A prognostic model for 30-day in-hospital mortality based on vital signs (systolic blood pressure, respiratory rate, and mental status) was compared with an extended model that also included four inflammatory markers (C-reactive protein, neutrophil-lymphocyte ratio, mean platelet volume, and red cell distribution width). Similar to the previous finding, all inflammatory markers except C-reactive protein showed significant contributions. Subsequently, a prediction model was developed using vital signs and markers with significant added value using a dataset that combined the external validation data with the data of the previous study. The new model was compared with a model based on the quick Sequential (sepsis-related) Organ Failure Assessment (qSOFA) score. The newly developed model showed a higher c-index than the qSOFA model [0.756 (95% CI 0.726-0.786) vs. 0.663 (0.630-0.696), p < 0.001]. Using the new model, 9.0% of patients who died were correctly reclassified compared with the qSOFA model at the threshold of 10% mortality risk. The new model including these markers showed potential to outperform the qSOFA model.

A paper-based colorimetric point-of-care testing kit using novel two-dimensional CuBO2 nanoplates for the efficient detection of Vitamin C and fabrication of a highly sensitive optical sensor

This work explores the development of a colorimetric on-spot testing kit and the fabrication of an optical sensor to efficiently detect Vitamin C(VC) using novel 2D-CuBO2 nanoplates, a vital marker for health and disease diagnostics. Although CuBO2 exhibits promising properties in various fields, including transparent conducting domains, its application toward sensing remains unexplored. Here, the authors report the primary utilization of 2D- CuBO2 as a potential candidate for sensing, specifically VC. The 2D-CuBO2 is synthesized through a one-pot sol-gel process followed by comprehensive characterization using various analytical techniques. The investigation of the effect of pH on the selective detection of VC was identified to be at a pH of 7. A paper-based sensor and device prototype for the on-spot detection is fabricated, calibrated, and analysed, showcasing the platform's sensitivity and a high degree of selectivity. The development of color can be attributed to the interaction of electromagnetic waves at the surface and interface of VC-modified aggregated CuBO2 nanoplates and, thereby, changes in the surface reflectivity leading to an observable color change. Optical sensing significantly improved the detection limit by several orders (LoD:130 nM) with a linear range of 10 μM to 10 mM.

Problems and level of satisfaction of patients during their stay in a tertiary care institute in Punjab

Patient satisfaction can be defined as the degree of congruency between a patient’s expectations of ideal care and his /her perception of the real care (s) he receives. It is a multidimensional aspect and represents a vital key marker for the quality of health care delivery.The cross-sectional study was conducted among the IPD patients in the indoor wards of SGRD Hospital which is a tertiary care hospital. A self-structured questionnaire was used to collect data. Questions regarding twelve potential problems were asked which could have been faced by the respondent during their stay in hospital.A total of 226 respondents were interviewed. 52.2% of the respondents were females and remaining (47.8%) were males. Overall satisfaction of the patients from the IPD services provided by the hospital was found to be 87.2%. Major problems faced by the respondent were difficulty in accessing the hospital (46%) and higher overall cost of treatment than expected (45.6%). Most of the respondents were satisfied with quality of food and water available (97.8%) and quality of linen provided by hospital and sanitary conditions of ward (97.3%).The study suggests that majority of In-patients are highly satisfied with the IPD services provided by the hospital which is highly motivating but every attempt must be made to form new policies and reform existing ones to achieve 100% satisfaction of patients. A number of potential barriers have been discovered in this study. Periodic patient satisfaction survey must be institutionalized in order to continuously improve hospital services. Patient counseling and health education cells could be open where patients and their relatives could solve their queries.

High eosinophil counts aid in diagnosing Kawasaki disease in febrile children

Abstract Background Several recent studies have validated the newfound significance of elevated eosinophils in Kawasaki disease (KD). However, this finding was not incorporated into the suggested laboratory criteria outlined by the Kawasaki Disease Committee of the American Heart Association for assessing children with incomplete KD. Purpose We aimed to determine the potential benefits of including eosinophil levels in the laboratory evaluation of KD. Methods A retrospective cohort study involved 77,934 febrile children (FC, not KD) and 1,970 subjects diagnosed with KD. Participants were recruited from four hospitals, comprising two medical centers and two regional hospitals. The AHA recommends specific laboratory tests—white blood cell count, hemoglobin, platelet count, ALT levels, albumin, and urinalysis—for evaluating incomplete KD in children with a higher CRP than 30mg/dL. The first four items are standard blood assays. Additionally, we introduced high eosinophil counts as a new diagnostic criterion for identifying children with KD. Results All these five laboratory findings (anemia for age defined as below the 5th percentile), platelet count &amp;gt;450,000/mm3, white blood cell&amp;gt;15,000/mm3, ALT&amp;gt;40 U/L, and eosinophil count &amp;gt;190/mm3) are significantly different between KD children and FC. The proportion of having these features is higher in KD children than in FC (19.3% vs. 8.0% in anemia for age, 20.5% vs. 10.3% in thrombocytosis, 37.0% vs. 18.1% in leukocytosis, 45.7% vs. 5.8% in elevated ALT, and 62.6% vs. 29.3% in high eosinophils). Elevated eosinophil counts are most commonly associated with KD among these five parameters. Furthermore, when incorporating the parameter of elevated eosinophil counts, the proportion of KD children with at least two abnormalities rose from 35.4% to 60.7%. In a subgroup analysis of children with KD presenting at least two abnormalities, the proportion rose from 38.7% to 63.5% in the high CRP group (≧ 3.0 mg/L) and 21.6% to 49.0% in the low CRP group (&amp;lt; 3.0 mg/L), respectively. Conclusions Higher eosinophil counts are vital markers in KD children. This aids in the identification of additional potential KD cases, especially in those exhibiting low CRP levels.

Multi-dimensional assessment of long-term outcomes in patients treated with extracorporeal cardiopulmonary resuscitation during refractory cardiac arrest

Abstract Background Recent randomised controlled trials showed short-term survival benefits with extracorporeal cardiopulmonary resuscitation (ECPR) compared to standard cardiac arrest treatment in selected patients with refractory cardiac arrest. Only limited data exist on multi-dimensional long-term outcomes after ECPR. Purpose To investigate multi-dimensional long-term outcomes of ECPR survivors, that is general health status, as well as cardiac, cognitive and psychological functions. Methods This is a multicenter, mixed-method clinical study conducted by cardiologists, neurologists and psychologists between May 2021 and December 2023. Adult patients who survived ECPR with veno-arterial extracorporeal membrane oxygenation (VA-ECMO) or "ECMELLA" (i.e., VA-ECMO plus percutaneous left-ventricular assist device) in two German tertiary care centers were included. A comprehensive set of functional outcomes across multiple dimensions were assessed at approximately 18 months after the index cardiac arrest event (see Figure for stations A to E and explanation of abbreviations). Results Long-term outcomes were assessed in 32 ECPR survivors, out of whom 15 patients (46%) had received VA-ECMO and 18 (54%) ECMELLA treatment. 23 patients (70%) were male and the average age was 55 years (±13). Vital signs and laboratory markers were unremarkable. Most patients denied heart failure symptoms although the average EQ-5D-5L score was 52 (±8). The left-ventricular ejection fraction was 51% (±9), which improved by 14% (±11) in the ECMO group and 31% (±15) in the ECMELLA group since the index cardiac arrest event. Bicycle exercise (on average 6 minutes duration with a power of up to 100 Watts) and 6-minute walk test (average distance of 394 meters ±145) revealed physical impairments. 29 patients (88%) had a good cerebral performance (i.e., Cerebral Performance Category scale of 1) and the average mental status was good (mini–mental state examination score 29 ±1). 22 patients (56%) perceived (partial) impairments in activities of daily living although the average Barthel index was 100 (±5) and modified Rankin scale was 1 (±1). 12 patients (36%) were unable to return to their jobs. Remarkably, 17 (51%) and 4 (12%) patients reported depressive symptoms and suicidal thoughts, respectively. Conclusion Although ECPR remains a highly complex procedure, cardiac and neurologic functions in survivors have almost recovered, while physical and psychological impairments still persisted at 18 months after cardiac arrest. Physical rehabilitation and psychological care appear to be important measures for ECPR survivors.

Presence of Plasmodium falciparum strains with artemisinin-resistant K13 mutation C469Y in Busia County, Western Kenya

Malaria remains a key health and economic problem, particularly in sub-Saharan Africa. The emergence of artemisinin drug resistance (ART-R) parasite strains poses a serious threat to the control and elimination of this scourge. This is because artemisinin-based combination therapies (ACTs) remain the first-line treatment in the majority of malaria-endemic regions in Sub-Saharan Africa. Certain single-nucleotide polymorphisms in the propeller domains of Plasmodium falciparum Kelch 13 protein (K13) have been associated with delayed parasite clearance in vivo and in vitro. These mutations serve as vital molecular markers for tracking the emergence and dispersion of resistance. Recently, there have been increasing reports of the emergence and spread of P. falciparum ART-R parasites in the Eastern Africa region. This necessitates continued surveillance to best inform mitigation efforts. This study investigated the presence of all reported mutations of K13 propeller domains in the parasite population in Busia County, Kenya, a known malaria-endemic region. Two hundred twenty-six participants with microscopically confirmed uncomplicated malaria were recruited for this study. They were treated with artemether–lumefantrine under observation for the first dose, and microscopic examination was repeated 1 day later after ensuring the participants had taken the second and third doses. P. falciparum DNA from all samples underwent targeted amplification of the K13 gene using a semi-nested PCR approach, followed by Sanger sequencing. The recently validated ART-R K13 mutation C469Y was identified in three samples. These three samples were among 63 samples with a low reduction in parasitemia on day 1, suggesting day 1 parasitemia reduction rate is a useful parameter to enrich the ART-R parasites for further analysis. Our findings highlight the need for continuous surveillance of ART-R in western Kenya and the region to determine the spread of ART-R and inform containment.

Exploring the repeatability of pulse arrival time in healthy subjects: A test-retest approach

Objectives: Vascular ageing is increasingly being recognised as a vital marker of cardiovascular morbidity and mortality. Assessment of vascular stiffness is an important parameter in this context. Pulse arrival time (PAT) assessed using photoplethysmography (PPG) and digital electrocardiogram (ECG) signals is a feasible and cost-effective parameter for this assessment. However, there are few, if any, studies that have assessed the test-retest repeatability of this parameter over time. Materials and Methods: We computed PAT using finger PPG and Lead II ECG and measured it sequentially at five instances over a period of 1 month in 21 healthy adults (10 males and 11 females). Mean and diastolic blood pressure (MBP and DBP) and heart rate (HR) were also measured at each visit. A novel parameter, PAT normalised for HR of 75 (PAT-75), was also computed. PAT and PAT-75 were compared for these visits using repeated measures analysis of variance. The intraclass correlation coefficient (ICC) was used to assess the test-retest reliability of this parameter. Results: MBP, DBP, and PAT values did not show any difference between the visits. HR was significantly different between the visits. PAT-75 was significantly lower for the afternoon of day 1 as compared to the forenoon. ICC demonstrated only moderate reliability of PAT (ICC = 0.57), with further reduction observed for PAT-75 (ICC = 0.38). Conclusion: PAT was only moderately repeatable on repeated evaluation over a 1-month period. This finding may have implications for the large-scale applicability of this technology, and therefore, we propose further investigation into the repeatability of this parameter in large cohorts.

The NRF2 inducer CDDO-2P-Im provokes a reduction in amyloid β levels in Alzheimer's disease model mice.

Alzheimer's disease (AD) is the most common aetiology of dementia. The transcription factor NF-E2-related factor 2 (NRF2) induces the expression of genes encoding phase II detoxification and antioxidant genes. NRF2 is regulated by Kelch-like ECH-associated protein 1 (KEAP1), and the KEAP1-NRF2 system is the key regulatory system involved in cytoprotection. To examine whether pharmacological induction of NRF2 expression alleviates AD phenotypes in vivo, we employed two AD mouse models, i.e. App NL-G-F/NL-G-F (AppNLGF) and APPV717I::TAUP301L (APP/TAU) mice. As the synthetic oleanane triterpenoid 1-[2-cyano-3,12-dioxooleana-1,9(11-dien-28-oyl)] (CDDO)-4(-pyridin-2-yl)-imidazole (CDDO-2P-Im) exhibits strong NRF2-inducing activity, we treated AD model mice with CDDO-2P-Im. We found that Aβ42 levels were markedly greater in the brains of AppNLGF mice than in those of APP/TAU mice. CDDO-2P-Im treatment significantly decreased Aβ42 levels, but not Aβ40 levels, in APP/TAU mice. Consequently, CDDO-2P-Im also decreased the ratio of Aβ42/Aβ40, a vital marker of amyloid plaque formation. LC-MS/MS analyses revealed that CDDO-2P-Im was delivered to the brains of the APP/TAU mice. CDDO-2P-Im induced the expression of detoxification and antioxidant gene targets of NRF2 and elevated reduced glutathione (GSH) levels in the mouse brain. These results support the notion that CDDO-2P-Im ameliorates AD-related pathologic changes.

OXIDATIVE STRESS MARKERS IN WESTAFRICAN DWARF AND KALAWAD GOATS AS AFFECTED BY THE EARLY RAINY SEASON

Superoxide dismutase (SOD) and malondialdehyde (MDA) are vital oxidative stress markers involved in regulating metabolism, immunity and homeostasis in animals to maintain health and productivity across changing environment. Thus, this study was undertaken to evaluate oxidative stress markers in westafrican dwarf and kalawad goats as affected by the early rainy season. The SOD and MDA levels of 16 non-gravid does consisting of eight West African Dwarf and eight KALAWAD goats managed intensively for a period of three months (May, June, and July), during the early rainy (hot-wet) season was investigated. Animals were fed Pennisetum purpureum and supplemented with concentrate at 2% body weight dry matter basis. Blood samples were collected twice a month for serum hormonal assay using ELISA kits. Data were analyzed using method of least square analysis of variance in a 2×3 factorial arrangement with the effects of breed and month of sampling as the independent variables while the SOD and MDA levels are the dependent variables. Results showed that breed had no significant effects (P&gt;0.05) on both SOD and MDA levels. Superoxide dismutase level decreases as the month of sampling progressed with May having the highest mean value followed by June and July with the lowest mean values respectively (2.41±0.58 vs 0.62±0.12 vs 0.60±0.16 U/L). The study concluded that SOD was higher in the two breeds in the month of May but the highest value was recorded in the KALAWAD goats which indicate that both breeds have high antioxidant levels with the KALAWAD tending to have the highest.

AI-driven transcriptomic encoders: From explainable models to accurate, sample-independent cancer diagnostics

In the rapidly evolving domain of medical technology, the utilization of sophisticated algorithms for deciphering transcriptional data has emerged as a critical aspect, especially in the oncology sector. These algorithms, drawing upon methodologies from fields such as natural language processing and advanced image analysis, can significantly enhance the accuracy in predicting cancer-related molecular states. Notably, Transformer models, renowned for their proficiency in handling extensive datasets, are now being adapted for breakthroughs in medical diagnostics or in stratifying patients according to prognostic levels. Our study contributes to the field of precision medicine by integrating Transformer-based learning, exemplified by the Geneformer model, with explainable AI techniques. These techniques are employed to find out the input variables (genes resulting from genomic transcription) most correlated with the decisions of neural network systems. This insight, a key goal in genomic research, aims to select the most relevant gene subset for each specific task in which a neural network is employed. This selection approach has proven to be effective in two classification tasks: cell type classification and breast cancer type classification. Such effectiveness has been demonstrated even across various cohorts of patients. When applying Geneformer-like architecture analyses solely to the selected gene subsets, the outcomes either maintain their accuracy or significantly improve. This approach, aims not only to contribute to the identification of vital genetic markers in cancer genomics, but also to exemplify the adaptability of AI models to different datasets, marking a significant step towards the development of accurate and universally applicable diagnostic tools for precision medicine.

From Waste to Sensor: Facile Synthesis of a Copper‐Doped Hydroxyapatite Nanocomposite as a Colorimetric Sensing Platform for Uric Acid

AbstractUric acid serves as a vital diagnostic marker for diseases like arthritis, gout, etc. Accurate quantification of UA is crucial for diagnosis. The hydroxyapatite (HAp), being catalytic in nature, was used as a template for copper nanoparticles (CuNPs) synthesis and for providing surface area for chemical reactions needed for sensing uric acid. HAp was synthesized from waste chicken bones, followed by doping with copper metal salt through calcination. The synthesis of copper hydroxyapatite (Cu‐HAp) nanocomposite was confirmed using different techniques. SEM displayed the nanoporous morphology of the material. The hexagonal crystal structure of HAp was revealed by X‐ray diffraction analysis, and energy dispersive X‐ray analysis showed the successful doping of Cu on HAp. The Cu‐HAp nanocomposite and 3,3′,5,5′‐tetramethylbenzidine (TMB) dye were employed as a new colorimetric sensing platform for uric acid sensing. The fabricated sensing platform detected uric acid with excellent sensitivity (LOD, 0.24 μM) and a wide linear range (1–270 μM). Optimization experiments demonstrated that the proposed sensor performs best at 4 mg of Cu‐HAp nanocomposite, pH 6, with a response time of 100 s. Furthermore, TMB and H2O2 concentrations of 12 and 8 mM were used, respectively. The proposed sensor successfully detected uric acid in physiological samples.

Alternative Strategy to Analyze In Vitro Cell Invasion of 3D Cultures.

Spheroid culture is a 3D model that provides an improved replication of the in vivo microenvironment compared to traditional two-dimensional (2D) cultures. Invasion is a cellular outcome of utmost interest in cancer biology. In this protocol, we have devised an alternative strategy for evaluating cancer cell invasion in vitro, employing heterospheroids comprised of oral squamous cell carcinoma (OSCC), cancer-associated fibroblasts (CAF), and monocytes. These heterospheroids aim to mimic the tumor microenvironment (TME), including two relevant non-neoplastic cell types alongside the cancer cells. Each cell type was labeled with vital fluorescent markers emitting in distinct wavelengths before spheroid formation. Once formed, heterospheroids were seeded onto a layer of human leiomyoma-derived extracellular matrix in the upper compartment of a microporous membrane. Invasion was assessed in the z-axis using confocal microscopy. Digital images were obtained in the corresponding fluorescent channels at 10 µm intervals, covering a depth of 90 µm in the z-axis. Analysis was performed using freeware image software by calculating the integrated fluorescence intensity in each image and fluorescence channel. This approach enables a more dynamic analysis of cell invasion patterns in a multilayered context, as well as the examination of spatial co-localization of different cell types during invasion.

15N metabolic labeling-TMT multiplexing approach to facilitate the quantitation of glycopeptides derived from cell lines.

The study of glycoproteomics presents a set of unique challenges, primarily due to the low abundance of glycopeptides and their intricate heterogeneity, which is specific to each site. Glycoproteins play a crucial role in numerous biological functions, including cell signaling, adhesion, and intercellular communication, and are increasingly recognized as vital markers in the diagnosis and study of various diseases. Consequently, a quantitative approach to glycopeptide research is essential. One effective strategy to address this need is the use of multiplex glycopeptide labeling. By harnessing the synergies of 15N metabolic labeling via the isotopic detection of amino sugars with glutamine (IDAWG) technique for glycan parts and tandem mass tag (TMT)pro labeling for peptide backbones, we have developed a method that allows for the accurate quantification and comparison of multiple samples simultaneously. The adoption of the liquid chromatography-synchronous precursor selection (LC-SPS-MS3) technique minimizes fragmentation interference, enhancing data reliability, as shown by a 97% TMT labeling efficiency. This method allows for detailed, high-throughput analysis of 32 diverse samples from 231BR cell lines, using both 14N and 15N glycopeptides at a 1:1 ratio. A key component of our methodology was the precise correction for isotope and TMTpro distortions, significantly improving quantification accuracy to less than 5% distortion. This breakthrough enhances the efficiency and accuracy of glycoproteomic studies, increasing our understanding of glycoproteins in health and disease. Its applicability to various cancer cell types sets a new standard in quantitative glycoproteomics, enabling deeper investigation into glycopeptide profiles.

A study on the utility of microalbuminuria as an indicator for the assessment of severity of chronic obstructive pulmonary disease and its co-morbidities

Abstract Background: Microalbuminuria (MAB) in chronic obstructive pulmonary disease (COPD) reflects generalised endothelial dysfunction as well as systemic inflammation. It is considered a vital marker of early cardiovascular involvement and other systemic involvement in COPD. MAB can serve as a surrogate marker to predict the severity of the disease and other systemic co-morbidities amongst COPD patients. This study was carried out to study the association between MAB with COPD severity and to compare the presence of MAB in COPD patients with and without co-morbidities. Methods: The study included 106 COPD patients diagnosed as per the Global Initiative for Obstructive Lung Disease (GOLD) criteria. Severity was assessed with the help of GOLD staging and modified Medical Research Council Dyspnoea Scale. MAB was assessed by urine albumin–creatinine ratio (UACR) of the spot urinary sample. The presence of comorbidities assessed by history, physical examination, laboratory investigations. Results: MAB was evident in n/N (70.8%) patients. MAB was found in 80.6%, 79.1%, 38.1% and 0% of the patients with GOLD Stage 4, Stage 3, Stage 2 and Stage 1 COPD patients, respectively (P = 0.0001). The mean UACR was significantly higher in severe stages of COPD. A significant association of MAB with co-morbidities such as hypertension, ishaemic heart disease, dyslipidaemia and diabetes mellitus (P &lt; 0.05) was evident. Conclusions: MAB could be a promising biomarker to identify COPD patients with severe disease and who are at risk of developing cardiovascular and other systemic co-morbidities.

Pediatric tuberculosis in a high burden setting: Socio-spatial distribution, blood elements features, and outcomes

BackgroundThe childhood tuberculosis (TB) epidemic has been long neglected. Data on pediatric tuberculosis is needed to develop effective strategies against TB. MethodsWe retrospectively reviewed 200 medical records from children aged 0–15 years who suffered from tuberculosis between 2011 and 2021 in Libreville, Gabon. We collected and analyzed socio-demographic data and clinical data. Results141 children files were selected (43 % girls and 57 % boys). The mean age of the patients was 9.2 years (CI: 8.5–10). Sixty per cent (60 %) of cases were from precarious housing areas, 35.34 % from mixed housing areas, and 4.51 % from residential. The cure rate was 75.24 %, 9.52 % relapsed, and 15.24 % died. Deaths were significantly higher in older children (Dunn's post-test p < 0.01). Children who recovered had higher haemoglobin and platelet counts than those who died (Dunn's test: haemoglobin p < 0.0001; thrombocytes p < 0.05). The haemoglobin threshold value of 5.5 g/dL identified children death with up to 80 % sensitivity and 86 % specificity. Thrombocytes count identified children's death with a sensitivity of 80 % and a specificity of 51 %. ConclusionPrecariousness is associated with childhood tuberculosis. The directly observed therapy (DOTS) in older children should be reinforced to limit tuberculosis-associated deaths. Haemoglobin concentration and platelet are vital prognosis markers in pediatric tuberculosis.