Analysis Of Markers Research Articles

Multi-Omic Characterization of Single Cells and Cell-Free Components Detected in the Cerebrospinal Fluid of Patients with Leptomeningeal Disease

Background/Objectives: Up to 30% of patients with breast cancers will develop brain or leptomeningeal metastases, and this risk is especially high with HER2-positive cancers. For patients with central nervous system metastases, cerebrospinal fluid (CSF) liquid biopsies are a promising opportunity to monitor disease, inform treatment, and predict prognosis. This pilot study investigated CSF liquid biopsy analytes from three patients diagnosed with central nervous system metastases based on imaging but not confirmed via clinical cytology. Methods: The detection of cellular analytes with the non-enrichment high-definition single-cell assay (HDSCA3.0) workflow was compared between the CSF and matched peripheral blood (PB) samples. Results: Circulating tumor cells (CTCs) were detected in the CSF but not the PB and were subsequently molecularly characterized using single-cell genomics and targeted multiplexed proteomics to reveal a clonal population of phenotypically heterogeneous cells. There was a lack of concordance in the copy number alteration profiles between CTCs and cell-free DNA (cfDNA) in the CSF. Extracellular vesicle surface marker analysis in CSF revealed a prominent signal among tetraspanins (CD9/CD63/CD81), with CD81 exhibiting the highest signal across all patients. Conclusions: The data presented suggest that CSF could be a useful tool for diagnosing and assessing disease severity.

Identification of clonally expanded γδ T-cell populations during CAR-Tcell therapy.

Anti-CD19 Chimeric Antigen Receptor (CAR)-Tcell therapies have shown promise for treating B cell malignancies, but the clinical outcome is influenced by both the CAR-T product and the patient's immune system. The role of γδ Tcells in the context of CAR-T cell therapy remains poorly understood. This study investigates the transcriptional heterogeneity, clonal expansion and dynamics of γδ Tcells in patients undergoing anti-CD19 CAR-T cell therapy. Longitudinal single cell multi-omics analysis was performed on γδ Tcells from four patients receiving anti-CD19 CAR-T cell therapy. Single cell RNA-seq, antibody-based protein profiling (AbSeq) and full-length TCRγδ sequences revealed clonally expanded populations displaying plasticity in Tcell differentiation, and temporal dynamics of large clones, suggesting ongoing expansion and differentiation. Clonally expanded γδ Tcells had heterogeneous gene expression profiles, occupying seven transcriptionally distinct clusters. Analysis of chemokine markers indicated cluster-specific homing tendencies of circulating γδ Tcells to peripheral tissues. We found unexpectedly high frequencies of Vδ1 and Vδ3 cells in the blood with distinct gene and protein expression profiles. This analysis provides insights into the dynamic and heterogeneous nature of γδ Tcells following anti-CD19 CAR-T cell therapy, contributing valuable information for optimizing CAR-T cell therapies in B cell malignancies.

The impact of exercise intensity and duration for swim training-induced adaptations in cardiac structure and function in women with mild hypertension.

This study aimed to investigate the impact of swim training intensity and duration on cardiac structure and function in mildly hypertensive women. Sixty-two mildly hypertensive women were randomized to 15 weeks of either (1) high-intensity swimming (HIS, n = 21), (2) moderate-intensity swimming (MOD, n = 21) or (3) control (CON, n = 20). Training sessions occurred three times per week. Cardiac measurements were conducted using echocardiography pre- and post-intervention. Both the HIS and MOD groups demonstrated significant within-group increases in left ventricular mass: 7.3% [1.2; 13.2] (p = 0.02) for HIS and 6.2% [0.5; 11.8] (p = 0.03) for MOD. The MOD group also demonstrated a significant increase in left ventricular internal dimension at end-diastole by 2.4% [0.2; 4.6] (p = 0.03).Post-hoc analysis of diastolic function markers revealed reduced mitral valve A velocity in both HIS (-14% [-25; -3], p = 0.02) and MOD (-13% [-23; -3], p = 0.01), leading to increased mitral valve E/A ratios of 27% [10; 47] (p = 0.003) and 22% [5; 40] (p = 0.01), respectively. Additionally, only MOD demonstrated increased left atrial diameter of 4.9% [0.7; 9.1] (p =0.02). A significant time×group effect (p = 0.02) existed for global longitudinal strain, which increased by 1.6% [0.2; 3.0] (p = 0.03) in MOD only. In conclusion, swim training for 15 weeks increased left ventricular mass and improved markers of diastolic function in mildly hypertensive women. These independent of exercise intensity and duration in mildly hypertensive women.

Analysis of Serum and Synovial Inflammatory Markers in Periprosthetic Joint Infections: A Narrative Review.

Periprosthetic joint infection (PJI) is considered a rare but devastating complication after total joint arthroplasty (TJA). The problem lies in the fact that there is a paucity of "gold standard" diagnostic tests that make the diagnosis of PJI extremely challenging. Recently, there have been increasing evidence-based guidelines that have been introduced to standardise the approach to a patient with a suspected PJI. Diagnosing a case of PJI traditionally involves initial screening for elevated serum inflammation markers C-reactive protein (CRP) (mg/dL) and erythrocyte sedimentation rate (ESR), and aspiration remains the sole confirmatory investigation. However, several factors would affect the values of the aforementioned markers, such as gender, age, and the presence of inflammatory circumstances. Serum D-dimer that detects fibrinolytic activities during infection has high sensitivity, but the specificity was not persuasive as it would elevate during other conditions, such as venous thromboembolism. Therefore, there is also a need for a simultaneous and secondary marker. There are also several synovial biomarkers, including ESR, CRP, alpha-defensin, and synovial fluid leukocyte count and differential for the detection of PJI. In this narrative review, we want to sum up the serum and inflammatory markers that have been introduced so far for detecting PJI.

Analysis of Discourse Markers and Their Combinations in Nonnative Academic Writing

Analysis of Discourse Markers and Their Combinations in Nonnative Academic Writing

Association analyses of nutritional markers with Parkinson’s disease and Alzheimer’s disease

IntroductionParkinson’s disease (PD) and Alzheimer’s disease (AD) are common neurodegenerative diseases with multifaceted etiology. Nutritional and metabolic abnormalities are frequently implicated in PD and AD. In this observational study, we analyzed a series of nutritional markers, and aimed to understand their association with AD and PD risk. MethodsA total of 424 PD patients, 314 AD patients, and 388 healthy controls were included. Nutritional markers including Hemoglobin A1c, vitamin B12, folate, apolipoprotein B (ApoB), apolipoprotein A1 (ApoA1), low-density lipoprotein (LDL), high-density lipoprotein, triglyceride, total cholesterol (TC), uric acid and homocysteine (HCY) were measured. Significance for odds ratios examining was P < 0.0045 after bonferroni correction. ResultsMultifactor risk analysis showed that ApoB, LDL, and TC reduce PD risk, while HCY increase PD risk. ApoA1 and HCY are protective and risk factors for AD, respectively. ConclusionThe cross-sectional study demonstrates that HCY and lipid metabolism markers are associated with PD and AD risks. Our findings support the involvement of one-carbon metabolism and lipid metabolism disturbance in PD and AD.

Quality Assessment of Corydalis saxicola Bunting Using Quantitative Analysis of Multi-Components by Single Marker and Fingerprint Analysis.

Yan Huanglian is the whole grass and rhizome of Corydalis saxicola Bunting (CSB) of the Papaveraceae family. CSB has various pharmacological effects and is a promising ethnopharmaceutical. The chromatographic fingerprint of 16 batches of CSB samples was established in this study. Quantitative analysis of multi-components by single marker and the external standard method (ESM) was used to quantitatively assess seven constituents of CSB. Because of its high stability and availability, dehydrocavidine was used as a reference to determine the relative correction factors (RCFs) of columbamine, epiberberine, coptisine hydrochloride, palmatine chloride, berberine hydrochloride, and chelerythrine based on high-performance liquid chromatography. The calculated quantitative analysis of multi-components by single marker values is consistent with those obtained from the ESM, and the repeatability of the RCFs was demonstrated. For the first time, the quantitative analysis of multi-components by the single marker method has been established for the simultaneous determination of seven components, and it is demonstrated to be feasible and accurate for the quality evaluation of CSB combined with chromatographic fingerprint analysis. The combination of these methods could be used as a reference for quality control in Chinese medicine.

Population structure and markers analysis associated with powdery mildew resistance in pumpkin, Cucurbita pepo L

Pumpkin (Cucurbita pepo L.) is an economically important vegetable and summer crop. The seeds have high nutritional value due to their edible oil and protein content with a high medicinal value. Pumpkin is attacked by several fungal diseases; one of which is powdery mildew (PM) caused by Podosphaera xanthii race-0. PM causes severe disease on pumpkin leading to major yield losses globally. In this study, we analyzed the genetic diversity and population structure of fourteen local pumpkin cultivars, all under Cucurbita pepo L. based on dominance in the region using 15 Inter-Simple-Sequence-Repeats-(ISSR) primer-pairs in association with resistance against PM as well as regulating the expression of three resistant marker genes i.e. SGT1, bHLH and PSK via qRT-PCR at three stages with 10 days' interval post-inoculation-stages-(SPI) with the powdery mildew agent, P. xanthii race-0. The results showed that ISSR markers demonstrated 13 primers out of 15 primers generated 169 distinct amplified bands, out of which 143 bands showed polymorphism. The mean polymorphism information content (PIC), marker index (MI), effective-number of alleles (I), expected heterozygosity (H), and Nei's gene diversity parameters were 0.354, 3.344, 0.334, 0.205 and 0.200, respectively. According to PIC, MI, I, H, and Nei indices evaluation, ISSR10 primer had the best performance in genetic diversity of the related pumpkin population. NTSYS pc 2.02e software and several similarity coefficients (r = 0.964), placed the related genotypes into five distinct groups. Of which, ‘Maragheh1′ and ‘Shahreza’ as resistant ones with 9.4 % and 6.1 % disease severity (DS), and ‘Gorgan’, ‘Urmia1′ and 3 and ‘Maragheh3’ with 79.6–84.8 % DS, as susceptible ones into two distinct groups, respectively. Further, the marker genes SGT1, bHLH and PSK expression level changes showed that the effect of variable factors including different genotypes (susceptible and resistant), disease-causing fungus, SPI, and the mutual effect of these factors is significant with the highest increase in resistant genotypes. Considering the relative compatibility in the classification of pumpkin genotypes in the dendrograms of DS and ISSR markers, it can be expected that increasing the number of markers in evaluating the genetic diversity of these genotypes makes it possible to differentiate pumpkin genotypes resistant and susceptible to PM disease based on the degree of disease resistance. The markers associated with resistance are recommended for use in marker assisted breeding.

THYROID DYSFUNCTION IN PATIENTS WITH TYPE 1 DIABETES AT A TERTIARY CARE HOSPITAL

Individuals with diabetes mellitus (DM) are reporting thyroid dysfunction at an alarming rate, and there's evidence that thyroid dysfunction—whether overt or subclinical—affects overall glycaemic control. 12.3% of individuals with Type 1 DM have overt hypothyroidism, while around 24.8% of patients have abnormal thyroid autoantibody levels. Objective: This study aimed to determine Thyroid dysfunction in patients with type 1 diabetes at a tertiary care hospital. Methods: The present descriptive study was carried out at Hayatabad Medical Complex Peshawar from January 2024 to June 2024 after obtaining permission from the ethical committee of the institute. A total of 110 participants were enrolled in the study. We included all T1DM patients of both genders. All participants with T1DM visiting the OPD were screened. The primary test for assessing thyroid function was TSH, which has an acceptable range from 0.5 to 5 mIU/ml. Triiodothyronine (T3) and free thyroxin (FT4) levels were also measured. On a Cobos 6000 machine (Roche), thyroid function tests (TFTs) were performed. Data was analyzed through SPSS version 22.0. Results: A total of 110 participants of both genders of different age groups were enrolled in this study. 63.63% of them were females and 36.36% were males. 18 individuals (16.36%) had subclinical hypothyroidism, 14 individuals (12.7%) were hypothyroid, and 78 of them (70.90%) were euthyroid. We did not identify any occurrences of hyperthyroidism in current research. In comparison to the rest of the population, individuals with hypothyroidism had significantly different mean Thyroid stimulating hormone, FT4 and FT3 levels (p &lt;0.0001). The mean differences for age, the period of diabetes, randomized blood sugar (RBS), and HbA1c were not statistically significant. When the data was examined for subclinical hypothyroidism, however, there was a substantial variance for mean Thyroid stimulating hormone (p &lt;0.0001) but not for FT4 or FT3. Likewise, there was no difference seen in age (p= 0.35) or gender (p= 0.63). Conclusion: Type 1 diabetes individuals often suffer from thyroid dysfunctions particularly hypothyroidism (12.7%) and subclinical hypothyroidism (16.36%). The analysis of thyroid-stimulating hormone levels and other hormonal markers for thyroid function is the most crucial approach.

Species Differentiation of Prunus serrulata and Prunus xueluoensis Based on Combined Analysis of SSR and cpDNA Markers

Species Differentiation of Prunus serrulata and Prunus xueluoensis Based on Combined Analysis of SSR and cpDNA Markers

Инфузия субанестетических доз кетамина в послеоперационном периоде как средство церебропротекции у детей при хирургической коррекции врожденных пороков сердца: проспективное рандомизированное исследование

INTRODUCTION: The world literature describes the cerebroprotective properties of ketamine when used in minimal doses, which is due to its effect on NMDA receptors and inhibition of damage to the neurovascular unit. However, there is a lack of research on the use of ketamine for cerebroprotection in pediatric populations and cardiac surgery. OBJECTIVES: To establish the safety and effectiveness of ketamine infusion for cerebroprotection in children in the postoperative period during cardiac surgery. MATERIAL AND METHODS: The study included 45 children aged from 1 to 60 months and weighing from 3.5 to 20 kg, who underwent surgical correction of atrial or ventricular septal defect of the heart under artificial circulation. Patients were randomized into a study group or a control group. In the study group, upon completion, patients received a ketamine infusion at a dose of 0.1 mg/kg/hour for 16 h. To establish the effectiveness of cerebroprotection, serum markers of damage to the neurovascular unit were used: protein S100-β, neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP), occludin, claudin-1. Blood sampling for marker analysis was carried out at three control points: before the start of the operation, immediately after completion of artificial circulation and 16 hours after completion of the operation. RESULTS: Al studied factors in the intra- and postoperative period did not differ significantly between groups. When analyzing markers of damage to a neurovascular unit, their maximum concentration was revealed at the second control point. For the NSE marker, a lower level was found in the group using ketamine — 16.52 [11.61–18.8] and 21.37 [17.78–28.74] ng/ml (p = 0.0019). CONCLUSIONS: The safety of using ketamine infusion in the postoperative period at a dose of 0.1 mg/kg/hour was revealed. Among all markers used, NSE serum concentrations were statistically significantly lower in the ketamine group.

Assessment of conservation status of Ferula huber‐morathii: association with population genetic structure and regional climate

Ferula huber‐morathii is an endemic and medicinally important plant. This species is distributed in eight Turkish localities, including three newly identified ones. Its extent of occurrence and area of occupancy is determined to be 3963 km2 and 32 km2 respectively. All localities are characterized by East Mediterranean and sub‐Mediterranean precipitation regimes; however, temperatures increase excessively and precipitation decreases during the flowering period of the species. The population sizes are quite small, and the number of reproducing individuals in some populations is below ten. Analyses of ISSR markers showed the percentage of polymorphic loci to be 94% at the species level and 56% at the population level. The level of genetic differentiation (measured by GST) was 0.37 and the estimated level of gene flow among populations (NM) was 0.84. The percentage of variance occurring within and among populations, determined by AMOVA, was 75% and 25%, respectively. STRUCTURE analysis revealed two genetic clusters of individuals with a geographic structure, similar to that found in UPGMA and an ordination analysis. Some populations turned out to have both low numbers of individuals and low genetic diversity. Since many of the populations are subject to anthropogenic disturbance, the species should remain in the EN category. At the same time, it is suggested that a new in‐situ conservation area should be created around nearby dams, situated in the same climate area as the currently known populations.

Amelioration of Systemic Amyloidosis by Blocking IL-17A and Not by IL-17F, and Arteriosclerosis by Blocking Both IL-17A and IL-17F in an Inflammatory Skin Mouse Model.

There are comorbidities and complications in atopic dermatitis and psoriasis that often occur after the appearance of skin inflammation. Statistically, data show that patients with psoriasis and atopic dermatitis have a shorter life expectancy than patients without psoriatic dermatitis, due to the occurrence of arteriosclerosis, myocardial infarction, and cerebral infarction. Many types of skin inflammation are treated with various antibody preparations, and marked improvement in patients' quality of life can be achieved. The next theme is to understand the pathogenesis of arteriosclerosis, myocardial infarction, stroke, and other complications associated with dermatitis and to find treatments and drugs to reduce their occurrence. The skin, a crucial immune organ, generates large amounts of inflammatory cytokines in response to various stimuli, leading to systemic inflammation and potential damage to internal organs. The link between inflammatory skin conditions like psoriasis and atopic dermatitis with serious health complications such as vascular disorders and systemic amyloidosis has been increasingly recognized. In psoriasis, biological treatments targeting Interleukin (IL)-17A, a key cytokine, have shown promise in reducing cardiovascular risks. Recent developments include treatments that target both IL-17A and IL-17F in the psoriasis field, though each cytokine's impact on internal organ damage is still under debate. Among visceral complications secondary to dermatitis, systemic amyloidosis and atherosclerosis have been reported to be controlled by suppressing IL-17 in the early stages of dermatitis. Still, it remains unclear whether suppressing IL-17 prevents organ damage in the late stages of persistent severe dermatitis. A study using a long-lasting dermatitis mouse model that overexpressed human caspase-1 in keratinocytes (Kcasp1Tg) investigated the effects of deleting IL-17A and IL-17F on visceral complications. Cross-mating Kcasp1Tg with IL-17A-, IL-17F-, and IL-17AF-deficient mice assessed the skin and visceral organs histologically, and RT-PCR analysis of aortic sclerosis markers was performed. Despite less improvement in dermatitis, deletion of IL-17A in Kcasp1Tg mice showed promising results in reducing multiple organ amyloidosis. On the other hand, the effect was observed in both IL-17A and IL-17F deleted mice for aortic sclerosis. The inhibition of IL-17A and IL-17F was suggested to reduce the risk of developing comorbidities in internal organs. IL-17A and IL-17F were found to act similarly or produce very different results, depending on the organ.

The efficacy of infrared diode laser in enhancing the regenerative potential of human periodontal ligament stem cells (hPDLSCs)

BackgroundThe present study aimed to investigate the effects of infrared diode laser irradiation on the proliferation and differentiation capacity of periodontal ligament stem cells (hPDLSCs), which are optimal cell sources for periodontal regeneration.MethodshPDLSCs were isolated and characterized by flow cytometric analysis of mesenchymal stem cell markers, and their trilineage differentiation capacity was tested. hPDLSCs were then cultured and irradiated with infrared diode laser (970 nm) at a power of 200 mW and a fluence of 4 J/cm2 for 3 s. MTT assay was performed to assess cellular proliferation. Cell cycle analysis was performed, and the impact of infrared diode laser irradiation on the stemness and osteogenic differentiation potential of hPDLSCs was evaluated via RT‒PCR.ResultsInfrared diode laser application enhanced the stemness, viability, proliferation, and differentiation of PDLSCs. Stem cell markers (OCT4, SOX2, and NANOG) were significantly upregulated in hPDLSCs exposed to laser irradiation. There was significant overexpression of RUNX2, ALP, OPN, and OCN on day 14 after laser application.ConclusionsThese findings provide valuable insights into the specific applications of infrared diode lasers to effectively regenerate periodontal tissues. The results can aid in the development of precise clinical protocols aimed at enhancing osseointegration and promoting tissue regeneration. Ultimately, the combination of infrared diode laser with hPDLSCs is promising for stimulating periodontal regeneration.

First De Novo genome assembly and characterization of Gaultheria prostrata.

Gaultheria Kalm ex L. (Ericaceae), a type of evergreen shrub, known as a natural source of methyl salicylate, possesses rich germplasm resources, strong habitat adaptability, significant ornamental value, and noteworthy pharmacological activities. However, due to the paucity of whole genomic information, genetically deep research in these areas remains limited. Consequently, we intend to obtain genome data through high-throughput sequencing, gene annotation, flow cytometry, transcription factors prediction and genetic marker analysis for a representative species of this genus, with Gaultheria prostrata selected for our study. In this study, we preliminarily obtained the genome of G. prostrata through next-generation sequencing methods. Utilizing 47.94 Gb of high-quality sequence data (108.95× coverage), assembled into 114,436 scaffolds, with an N50 length of 33,667 bp. The genome size assembled by SOAPdenovo, approximately 417 Mb, corresponded closely to predictions by flow cytometry (440 Mb) and k-mer analysis (447 Mb). The genome integrity was evaluated using BUSCO with 91%. The heterozygosity ratio was 0.159%, the GC content was 38.85%, and the repetitive regions encompassed over 34.6% of the genome. A total of 26,497 protein-coding genes have been predicted and annotated across Nr, Swissprot, GO, KEGG, and Pfam databases. Among these, 14,377 and 2,387 genes received functional annotation in Nr and Swissprot, respectively; 21,895, 24,424, and 22,330 genes were similarly annotated in GO, KEGG, and Pfam. Moreover, A total of 279,785 SSRs were identified and 345,270 primers for these SSRs were designed. Within the various nucleotide types of SSRs, AG/CT and AAG/CTT constituted the predominant dinucleotide and trinucleotide repeat types in G. prostrata. In addition, 1,395 transcription factors (TFs) from 75 TF families, 462 transcription regulators (TRs) from 33 TR families and 840 protein kinase (PKs) from 118 PK families were identified in this genome. We also performed phylogenetic analyses of G. prostrata and related species, including estimation of divergence times and expansion and contraction analyses, followed by positive selection analyses of orthologous gene pairs of G. prostrata and its close relative Vaccinium corymbosum. These results provide a reference for in-depth study of genus Gaultheria, contributing to future functional and comparative genomics analyses and providing supporting data for the development of molecular markers.

Machine learning methods for diagnostic disease prediction and treatment decisions in parvovirus B19 positive viral cardiomyopathy

Abstract Background Parvovirus B19 (B19V) is the most common type of virus found in endomyocardial biopsies (EMB) from patients with inflammatory heart disease. However, the detection of genomic B19V-DNA appears to have little clinical significance and is often considered an incidental finding. Recent analyses of a large cohort of B19V-positive patients revealed that the detection of RNA transcripts rather than DNA is of clinical importance, as B19V-RNA-positive patients have a significantly worse cardiovascular outcome. Here, we applied machine learning (ML) methods to assess the impact of B19V transcriptional activity on cardiac inflammation and patient clinical outcome. Methods and results A total of 305 patients with a known clinical course and a positive endomyocardial B19V-DNA test were included in this study. Follow-up measurements of the B19V-specific transcripts VP1/2 and NS1 revealed significantly worse clinical courses than in patients without B19V-RNA detection (P=0.019, hazard ratio (95% CI)=1.62 (1.10-2.57)). LVEF at follow-up was significantly lower in patients with detected B19V-RNA (P=0.007), and adverse events such as worsening LV function (P=0.003) or stable systolic dysfunction (P=0.013) were more frequent in this group. Importantly, analysis of inflammatory markers in EMB showed no difference in patients with and without RNA detection (P=0.432), suggesting that the deterioration in clinical outcome was related to B19V transcriptional activity rather than an increase in inflammation. The analysis of area under the receiver operating characteristic curve (AUROC) of B19V-RNA-negative patients revealed that the macrophage marker MAC-1 (AUROC=0.66, P=0.002) was the single most prognostically relevant parameter, while the T cell marker CD3 (AUROC=0.73, P&amp;lt;0.001) was identified as the most prognostically relevant marker in B19V-RNA-positive patients. The linear combination of several inflammatory markers increased the AUROC value to 0.67 in B19V-RNA-negative patients and to 0.74 in B19V-RNA-positive patients. To further increase prognostic accuracy, we trained ML models on features derived from EMB inflammatory markers. For the prediction of major adverse cardiac events within 60 months, the best ML-models achieved an AUROC of 0.87 in B19V-RNA-positive and of 0.86 in B19V-RNA-negative patients, while models that neglect the differentiation of patients with regard to the presence of B19V RNA transcripts only achieved an AUROC of up to 0.76. Conclusions These results show that B19V transcriptional activity is an independent risk marker for adverse cardiac events in patients with viral cardiomyopathy and thus confirm the clinical-therapeutic importance of myocardial biopsy. Furthermore, we present an ML-based algorithm that accurately indicates disease progression in B19V-positive cardiomyopathy patients and is thus helpful for appropriate therapy recommendation.

Assessment of Inflammatory Markers (CBC and CRP) and their Impact on Severity and Recurrence of Pneumonia in Under-Five Children

Background: Pneumonia remains a leading cause of morbidity and mortality in children under five. This study evaluates the association between inflammatory markers complete blood count (CBC) parameters and C-reactive protein (CRP) levels and pneumonia severity and recurrence. Methods: A prospective study was conducted with 110 children diagnosed with pneumonia at Dr. Sirajul Islam Medical College &amp; Hospital and Islami Bank Hospital, Motijheel, Dhaka, from January to December 2023. Clinical assessments included demographic data, pneumonia severity classification, and inflammatory marker measurements. Results: The study population comprised 62 males (56.4%) and 48 females (43.6%), with a mean age of 24.6 months. Inflammatory marker analysis revealed significant differences in WBC count, neutrophil percentage, lymphocyte percentage, and CRP levels across mild, moderate, and severe pneumonia groups (p &lt; 0.001). Higher levels of WBC count (14,500 ± 3,500), neutrophils (72.2 ± 8.6%), and CRP (38.5 ± 11.2) were observed in patients with pneumonia recurrence compared to those without recurrence. Multivariate regression analysis identified WBC count, neutrophil count, and CRP levels as significant predictors of pneumonia recurrence. Conclusion: Elevated inflammatory markers, particularly CBC parameters and CRP levels, correlate with increased severity and recurrence of pneumonia in young children. Monitoring these markers can aid in early identification and management of at-risk pediatric patients, potentially improving clinical outcomes. Further research is needed to explore additional biomarkers and their implications in pediatric pneumonia management.

The molecular anti-metastatic potential of CBD and THC from Lebanese Cannabis via apoptosis induction and alterations in autophagy

The medicinal plant Cannabis sativa L. (C. sativa) is currently being extensively studied to determine the full extent of its therapeutic pharmacological potential. Δ9-tetrahydocannabinol (THC) and cannabidiol (CBD) are the most thoroughly investigated compounds. We aimed to explore the anticancer activity of cannabinoids mixture isolated from the Lebanese C. sativa plant in ratios comparable to the local medicinal plant, to elucidate its mechanism of action in breast cancer cells in vitro. Cells were subjected to cytotoxicity assay, cell cycle analysis, Annexin V/PI dual staining, cell death ELISA, immunofluorescence, in addition to western blot analysis of apoptotic and autophagy markers. We further evaluated the anti-metastatic effect of cannabinoids on MDA-MB-231 using the scratch wound-healing, trans-well migration and invasion assays. Our results revealed the promising therapeutic benefits of CBD/THC on inhibiting the growth of breast cancer cells by promoting cellular fragmentation, phosphatidylserine translocation to the outer membrane leaflet and DNA fragmentation in both cell lines while inhibiting the motility of the triple negative breast cancer cells. In our study, CBD/THC mixture was found to exhibit a pro-apoptotic activity via the activation of the mitochondrial apoptotic pathway, independent from ROS production while also suggesting the activation of a caspase-dependent apoptotic pathway. Even though autophagy was altered upon exposure to the cannabinoid mixture, our data suggested that it is not the mechanism responsible of inducing cell death. In conclusion, our study demonstrates the promising therapeutic benefits of CBD and THC isolated from the Lebanese C. sativa plant on breast cancer cells in vitro.

Optimization of loop mediated isothermal amplification assay (LAMP) for detection of chloroquine resistance in P. vivax malaria

Chloroquine is still used as a first-line treatment for uncomplicated Plasmodium vivax malaria in India and resistance to this therapy can act as a major hurdle for malaria elimination. It is difficult to monitor drug-efficacy and drug resistance through in vivo and in vitro studies in case of Plasmodium vivax so analysis of molecular markers serves as an important tool to track resistance. Molecular methods that are currently in use for detecting single nucleotide polymorphisms in resistant genes including Polymerase chain reaction (PCR), Realtime-Polymerase chain reaction require highly sophisticated labs and are time consuming. So, with this background the study has been designed to optimize Loop Mediated Isothermal Amplification Assay to detect single nucleotide polymorphisms in chloroquine resistance gene of Plasmodium vivax in field settings. Eighty-eight Plasmodium vivax positive samples were collected. Pvmdr1 gene was amplified for all the samples and sequenced. Obtained sequences were analyzed for the presence of single nucleotide polymorphisms in the target gene. Further Loop Mediated Isothermal Amplification Assay primer sets were designed for the target mutants and the assay was optimized. Clinical as well as analytical sensitivity and specificity for the assay was calculated. Double mutants with variations at T958M and F1076L were detected in 100% of the Plasmodium vivax clinical isolates with haplotype M958 Y976 Y1028 L1076. Designed primers for Loop Mediated Isothermal Amplification Assay successfully detected both the mutants (T958M and F1076L) in 100% of the isolates and do not show cross-reactivity with other strains. So, the assay was 100% sensitive and specific for detecting single nucleotide polymorphisms in the target Pvmdr1 gene. Limit of detection was found to be 0.9 copies/µl and lowest DNA template concentration detected by designed assay was 1.5 ng/µL. Observed prevalence of single nucleotide polymorphisms in Pvmdr 1 gene is indicating a beginning of trend towards chloroquine resistance in Plasmodium vivax. The present study optimized LAMP for detecting single nucleotide polymorphisms in Plasmodium vivax cases in field settings, thus would help in finding significant hubs of emerging chloroquine drug resistance and ultimately helping in the management of suitable antimalarial drug policy.

Prognostic Significance of Tumor-Stroma Ratio (TSR) in Head and Neck Squamous Cell Carcinoma: Systematic Review and Meta-Analysis.

The management of head and neck squamous cell carcinoma (HNSCC) relies heavily on TNM staging and WHO histologic grading; however, in recent years, the analysis of prognostic markers expressed in the tumor stroma has gained attention. The tumor-stroma ratio (TSR) quantifies the proportion of tumor tissue relative to the surrounding stromal tissue; it is assessed with the percentage of stromal tissue within the tumor area, with a cutoff point of 50% being widely used to discriminate high-stroma cancer. In this systematic review and meta-analysis, we investigated the potential prognostic role of the TSR in HNSCC. After a literature screening, 24 studies dealing with the TSR and survival outcomes were included. The TSR showed a significant association with overall survival (OS) in both unadjusted and adjusted measures (RR 2.04, CI 1.57-2.65, p < 0.01; HR 2.36 CI 1.89-2.94, p < 0.00001), with an even stronger prognostic potential in oral cavity/oral tongue cancers (RR 2.44 CI 1.84-3.22, p < 0.00001). The TSR also showed prognostic value when dealing with cancer-specific survival and was associated with a reduction in disease-free survival (DFS). In particular, the TSR also retained its prognostic role in terms of DFS when specifically considering early-stage cancers in both unadjusted and adjusted analyses (RR 1.81 CI 1.57-2.10, p < 0.00001; HR 2.09 CI 1.58-2.76, p < 0.00001). Therefore, we conclude that the TSR is a reliable prognostic marker that is easy to assess in routine histological slides and can be effectively implemented in the routine evaluation of HNSCC.