Cell Count Research Articles

Assessment of non-myelotoxic agents as a preparatory regimen for hematopoietic stem cell gene therapy.

RAG2 deficiency is characterized by a lack of B and T lymphocytes, causing severe lethal infections. Currently, RAG2 deficiency is treated with a Hematopoietic Stem Cell transplantation (HSCT). Most conditioning regimens used before HSCT consist of alkylating myelotoxic agents with or without irradiation and affect growth and development of pediatric patients. Here, we developed a non-myelotoxic regimen using G-CSF, VLA-4I or AMD3100. These agents are known HSC mobilizers or affect bone marrow (BM) permeability and may support the homing of HSCs to the BM, without inducing major side effects. Female Rag2-/- mice were pre-treated with Busulfan (BU), G-CSF, VLA-4I or AMD3100 and transplanted with male BM cells transduced with a lentiviral vector carrying codon optimized human RAG2 (RAG2co). Peripheral blood cell counts increased significantly after G-CSF, VLA-4I and AMD3100 treatment, but not after BU. Reconstitution of PB lymphocytes was comparable for all groups with full immune reconstitution at 6months post transplantation, despite different methods of conditioning. Survival of mice pre-treated with non-myelotoxic agents was significantly higher than after BU treatment. Here, we show that the non-myelotoxic agents G-CSF, VLA-4I, and AMD3100 are highly effective as conditioning regimen before HSC gene therapy and can be used as an alternative to BU.

Role of Preoperative Inflammatory Blood Cell Indexes as a Postoperative Risk Predictor Among Patients Undergoing On-Pump Cardiac Surgery.

Estimating patient risk before heart surgery (HS) is crucial. Perioperative inflammation is associated with several complications and mortality. This study investigated blood cell count inflammatory indices (BCCII) to predict risks, including neutrophil-to-lymphocyte ratio (NLR), derivate NLR (DNLR), neutrophil-to-platelet-lymphocyte ratio (NLPR), lymphocyte-to-monocyte ratio, platelet-to-lymphocyte ratio (PLR), Systemic Inflammatory Index (SII), Systemic Inflammatory Reaction Index (SIRI), and Aggregate Index of Systemic Inflammation (AISI). Data from a cohort of patients undergoing on-pump HS at a single center in Brazil were retrospectively analyzed. Data were obtained from medical records and a laboratory analyzer, and SPSS version 20.0 was used for index calculations and statistical analyses. In total, 444 surgeries were performed, and 40 in-hospital deaths occurred. Except for PLR, all other indexes were independent predictors of death after multivariate adjustment (all p < 0.05). Discrimination performance was absent for PLR and AISI, and NLR, NLPR, and DNLR demonstrated the best area under the receiver operating characteristic curve (AUC > 0.7; all p < 0.0001). For survivors (n = 404), all indexes exhibited a correlation with the length of hospital stay (all p < 0.03), and NLR, NLPR, and DNLR were predictors (p < 0.026) of poor operative outcomes (acute myocardial infarction, cerebrovascular attack, cardiac arrest, low cardiac output, prolonged mechanical ventilation, renal failure, and sepsis). All BCCII scores were associated with length of hospital stay. Apart from PLR, all indexes were independent predictors of in-hospital mortality. Accuracy was highest for NLR, NLPR, and DNLR; for survivors, these three factors were good predictors of poor operative outcomes.

Comparative Evaluation of Antibiotic Therapies for Subclinical Staphylococcal Mastitis in Goats

Subclinical mastitis (SCM) is characterized by inflammation of the udder without visible signs of inflammation in the animal, udder and milk and hence could be diagnosed by laboratory examination only. Effective treatment of SCM in goats is crucial for maintaining quality milk production. This study was conducted at the Department of Veterinary Medicine, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur (M.P.), India. A total of 236 lactating goats irrespective of age, breed, parity and stage of lactation, from both organized and unorganized sectors in and around Jabalpur, were screened over a period of six month. Based on modified California mastitis test scores, 83 goats tested positive for subclinical mastitis resulting in an overall occurrence of 35.17% (83/236). A total of 48 milk samples that tested positive for modified California mastitis test were further analyzed for bacterial isolation and identification. Among them, 34 (70.83%) samples were positive for only Staphylococcus spp., while 12 (25%) samples exhibited mixed infection with Staphylococcus spp. and Gram-positive bacilli. Then, 18 goats, positive for subclinical Staphylococcal mastitis were divided into three groups i.e. G1, G2 and G3 with six goats in each group for therapeutic study and six apparently healthy goats were taken as healthy control group (G4). G1 Goats were treated with Inj. Enrofloxacin LA @ 7.5 mg/kg b. wt. I/M single dose; G2 with Inj. Ceftizoxime @ 10 mg/kg b. wt. I/V single dose and G3 with Inj. Oxytetracycline LA @ 20 mg/kg b. wt. I/M single dose. The responses of therapeutic assessment were evaluated on the basis of somatic cell count and modified California mastitis test score. The most effective response for subclinical Staphylococcal mastitis in goats was observed with Ceftizoxime followed by Enrofloxacin LA and the least with Oxytetracycline LA.

Therapeutic Efficacy of Eucalyptus Oil as Teat Dip in Subclinical Mastitis in Goats

Mastitis, an inflammation of the parenchyma of mammary gland, results in alterations to the chemical and physical composition of milk due to a variety of infectious and non-infectious causes. The present work was conducted in the Department of Veterinary Medicine, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur, Madhya Pradesh (M.P.). A total of 256 lactating goats irrespective of age, breed, parity and stage of lactation belonging to both organized and unorganized sectors at Jabalpur were screened from “February 2024 to July 2024”. On the basis of modified California mastitis test (MCMT) score, out of 256 lactating goats, 98 goats were found positive for subclinical mastitis with overall occurrence as 38.28% on animal basis and 33.33% on udder halves basis. Therapeutic study on 24 milk samples positive for subclinical mastitis by MCMT was conducted and categorized into four treatment groups (G1, G2, G3 and G4), each group comprised of 6 goats. Six apparently healthy goats were selected as healthy control group (G5). Two different teat dips viz. Eucalyptus oil 98% + DMSO 2% BID for 14 days in groups G1 and G3 and 5% Povidone iodine BID for 14 days in groups G2 and G4 were used as teat dips, respectively. Moreover, goats of groups G3 and G4 were commonly treated with parenteral antibiotic amoxicillin sulbactam @10 mg/kg b.wt. I/M OD for 3 days. Significantly higher values of somatic cell count, standard plate count, pH, freezing point and significantly lower value of lactose were observed in subclinical mastitis affected goats. The mean density and mean water percentage values varied within normal physiological range. On the basis of restoration of milk parameters and standard plate count, the most effective therapy was elicited by Eucalyptus oil 98%+ DMSO 2% as teat dip in combination with antibiotic amoxicillin sulbactam (G3).

Development of Technology for the Synthesis of Nanocrystalline Cerium Oxide Under Production Conditions with the Best Regenerative Activity and Biocompatibility for Further Creation of Wound-Healing Agents

Background/Objectives: The issue of effective wound healing remains highly relevant. The objective of the study is to develop an optimal method for the synthesis of nanosized cerium oxide powder obtained via the thermal decomposition of cerium carbonate precipitated from aqueous nitrate solution for the technical creation of new drugs in production conditions; the select modification of synthesis under different conditions based on the evaluation of the physicochemical characteristics of the obtained material and its biological activity, and an evaluation of the broad-spectrum effect on cells involved in the regeneration of skin structure as well as antimicrobial properties. Methods: Several modes of the industrial synthesis of cerium dioxide nanoparticles (NPs) were carried out. The synthesis stages and the chemical and physical parameters of the obtained NPs were described using transmission electron microscopy (TEM), X-ray diffraction, Raman spectroscopy, and mass spectrometry. The cell cultures of human fibroblasts and keratinocytes were cultured with different concentrations of different nanoceria variations, and the cytotoxicity and the metabolic and proliferative activity were investigated. An MTT test and cell counting were performed. The antimicrobial activity of CeO2 variations at a concentration of 0.1–0.0001 M against Pseudomonas aeruginosa was studied. Results: The purity of the synthesized nanoceria powders in all the batches was &gt;99.99%. According to TEM data, the size of the NPs varied from 1 nm to 70 nm under different conditions and methodologies. The most optimal technology for the synthesis of the nanoceria with the maximum biological effect was selected. A method for obtaining the most bioactive NPs of optimal size (up to 10 nm) was proposed. The repeatability of the results of the proposed method of nanoceria synthesis in terms of particle size was confirmed. It was proven that the more structural defects on the surface of the CeO2 crystal lattice, the higher the efficiency of the NPs due to oxygen vacancies. The strain provided the best redox activity and antioxidant properties of the nanoceria, which was demonstrated by better regenerative potential on various cell lines. The beneficial effect of synthesized nanoceria on the proliferative and metabolic activity of the cell lines involved in skin regeneration (human fibroblasts, human keratinocytes) was demonstrated. The antimicrobial effect of synthesized nanoceria on the culture of the most-resistant-to-modern-antibiotics microorganism Pseudomonas aeruginosa was confirmed. The optimal concentrations of the nanoceria to achieve the maximum biological effect were determined (10−3 M). Conclusions: It was possible to develop a method for the industrial synthesis of nanoceria, which can be used to produce drugs and medical devices containing CeO2 NPs.

Impact of Obesity Severity on Hepatic Steatosis and Systemic Inflammatory Markers: A Comparative Analysis Across Obesity Classes

Introduction: Obesity has become a global health issue, with its prevalence steadily increasing. It is closely linked to several metabolic disorders, cardiovascular diseases, non-alcoholic fatty liver disease, and chronic low-grade inflammation and can progress to more severe liver conditions. This study evaluates the relationship between obesity and inflammatory markers in individuals with different obesity levels. Methods: A cross-sectional study was conducted among 50 patients categorized into three obesity classes based on body mass index (BMI). Blood samples were taken to evaluate inflammatory and metabolic markers, including white blood cell (WBC) count, neutrophil-to-lymphocyte ratio (NLR), and C-reactive protein (CRP). Results: There were no statistically significant differences in inflammatory markers such as WBC, NLR, or CRP; a trend toward higher CRP levels was observed in Class 3 obesity. Conclusion: In our study, no statistically significant association was observed between inflammatory markers and the degree of obesity. Although the sample size was relatively small, it is essential to acknowledge that obesity is a multifaceted condition, and genetic variations may play a role in these results.

HIV replication and tuberculosis risk among people living with HIV in Europe: A multicohort analysis, 1983-2015.

HIV replication leads to a change in lymphocyte phenotypes that impairs immune protection against opportunistic infections. We examined current HIV replication as an independent risk factor for tuberculosis (TB). We included people living with HIV from 25 European cohorts 1983-2015. Individuals <16 years or with previous TB were excluded. Person-time was calculated from enrolment (baseline) to the date of TB diagnosis or last follow-up information. We used adjusted Poisson regression and general additive regression models. We included 272,548 people with a median follow-up of 5.9 years (interquartile range [IQR] 2.3-10.9). At baseline, the median CD4 cell count was 355 cells/μL (IQR 193-540) and the median HIV-RNA level 22,000 copies/mL (IQR 1,300-103,000). During 1,923,441 person-years of follow-up, 5,956 (2.2%) people developed TB. Overall, TB incidence was 3.1 per 1,000 person-years (95% confidence interval [CI] 3.02-3.18) and was four times higher in patients with HIV-RNA levels of 10,000 compared with levels <400 copies/mL in any CD4 stratum. CD4 and HIV-RNA time-updated analyses showed that the association between HIV-RNA and TB incidence was independent of CD4. The TB incidence rate ratio for people born in TB-endemic countries compared with those born in Europe was 1.8 (95% CI 1.5-2.2). Our results indicate that ongoing HIV replication (suboptimal HIV control) is an important risk factor for TB, independent of CD4 count. Those at highest risk of TB are people from TB-endemic countries. Close monitoring and TB preventive therapy for people with suboptimal HIV control is important.

Clinical significance of circulating long non-coding RNA SNHG1 in type 2 diabetes mellitus and its association with cell proliferation of pancreatic β-cell

BackgroundTo explore the association of long non-coding RNA (lncRNA) SNHG1/ miR-195 axis with type 2 diabetes mellitus (T2DM) and islet function.MethodsThe expression of SNHG1 and miR-195 was measured in T2DM patients and in healthy subjects. Correlation between indciators was evaluated using Pearson correlation analysis. INS-1 cells were used to perform the cell function assays. Insulin secretion by INS-1 was detected using ELISA. Cell counting kit-8 (CCK-8) and flow cytometry was used to detect cell proliferation and apoptosis. Luciferase report assay was to used to verify the target of SNHG1.ResultsThe expression of SNHG1 was increased and miR-195 level was decreased in the serum of T2DM patients. Both SNHG1 and miR-195 could be biomarkers for T2DM diagnosis. The fasting plasma glucose (FPG) and HbA1c were positively related to SNHG1 and negatively related to miR-195. SNHG1 inhibited insulin secretion, and cell proliferation and promoted apoptosis of INS-1 cells via binding to miR-195.ConclusionsDetection of SNHG1 and miR-195 might predict T2DM. SNHG1 could suppress proliferation and insulin secretion, but promote apoptosis of INS-1 cells via sponging miR-195.

Biofertilizer based on &lt;i&gt;Agrobacterium &lt;/i&gt;as a key to food security

Background: The concept of food security is contingent upon the fulfillment of two fundamental requirements: physiological and socioeconomic. The former comprises providing sustenance that is both safeguarded and enhanced in line with people’s nutritional needs and priorities, whereas the latter includes the mechanisms by which these needs are supplied. The objective is to ensure a state of robust and healthy well-being. As a sustainable solution, biofertilizers play a pivotal role in addressing the challenges of food security. Biofertilizers represent a cost-effective and environmentally friendly solution, with the potential to enhance soil productivity through nitrogen fixation. They also enhance crop yield by increasing the concentration of nutrients. Access to functional foods derived from biofertilizer-enhanced yields may contribute to improved dietary diversification and overall well-being. Objective: The objective is to isolate and screen the most active nitrogen-fixing bacterium as a basis to a new biofertilizer. Methods: The isolation of nitrogen-fixing bacteria was conducted using the soil sowing technique. The selection of nitrogen-fixing bacteria was based on their morphophysiological characteristics. Bacterial growth was evaluated by viable cell count. Cultivation of the selected bacterium occurred in both flasks (on a shaker) and a bioreactor to compare growth conditions. The stimulatory effect of the selected bacterium on seed germination was assessed based on the final germination rate. The selected bacterium identified through molecular taxonomy. Results: A total of 100 pure cultures were isolated, from which nitrogen-fixing bacteria were selected. The cultivation conditions for these bacteria were optimized regarding pH, temperature and process duration in a flask on a shaker. Strain NB4 exhibited the most favorable development under optimal settings. Additionally, cultivation parameters for this strain were optimized in a laboratory bioreactor. Notably, a bacterial liquid culture water solution comprising 1.5% strain NB4 facilitated complete germination of wheat (Triticum aestivum L.), seeds and produced high-quality sprouts. The 16S rRNA gene sequence of strain NB4 was submitted to the GenBank database under accession number MT670424.1, identifying it as Agrobacterium Pusense RP 1. Conclusion: This research aimed to develop an effective biofertilizer to tackle the pressing issue of food security. The project’s cornerstone was Agrobacterium pusense RP 1, nitrogen-fixing strain isolated and identified through rigorous research. Keywords: Isolation and identification of nitrogen-fixing bacteria, cultivation in bioreactor, germination of wheat seeds, Agrobacterium pusense.

Chlorine dioxide is a broad-spectrum disinfectant against Shiga toxin-producing Escherichia coli and Listeria monocytogenes in agricultural water

Agricultural water is commonly treated with chlorine-based disinfectants, which are impacted by water quality. Understanding how water quality influences disinfectants such as chlorine dioxide (ClO2) against pathogenic bacteria is important for creating efficacious sanitation regimens. In this study, the minimum inhibitory concentration (MIC) of ClO2 needed to achieve a 3-Log reduction against Shiga toxin-producing Escherichia coli (STEC) and Listeria monocytogenes was compared across agricultural water samples. Sterile ddH2O served as a control to compare with environmental samples from Salinas Valley, CA, and laboratory standards. To test different dosages and water qualities, stock ClO2 was diluted in 24-well plates with target concentrations of 10, 5, 2.5, and 1.25 mg/L. Well plates were inoculated with pathogens and treated with sanitizer for 5 min. Following treatment, surviving pathogens were enumerated using viable cell counts. The results demonstrate that groundwater samples had the highest water quality of the environmental samples and required the lowest concentration of disinfectant to achieve 3-Log reduction against both bacteria, with MIC between 1.4 and 2.0 mg/L. Open-source samples had lower water quality and required a higher concentration of ClO2 for 3-Log reduction, with MIC between 2.8 and 5.8 mg/L for both pathogens. There was no correlation between pH, turbidity, or conductivity/TDS and reduction for either STEC or L. monocytogenes, suggesting no individual water metric was driving reduction. A lower dosage was required to achieve 3-Log reduction against STEC, while L. monocytogenes required greater concentrations to achieve the same level of reduction. Overall, these results help guide growers in using ClO2 as a broad-spectrum disinfectant and demonstrate its efficacy in reaching 3-Log reduction across agricultural water samples.

Allicin Ameliorated High-glucose Peritoneal Dialysis Solution-induced Peritoneal Fibrosis in Rats via the JAK2/STAT3 Signaling Pathway.

Peritoneal fibrosis (PF) is one of the most serious complications of peritoneal dialysis (PD) and is the greatest obstacle to the clinical application of PD. Chinese herbal monomers have been effective in the prevention and treatment of PF. The aim of this study was to observe the effect of allicin on PF in rats induced by high glucose and to investigate its molecular mechanism of action. A rat model of PF was established by using a 4.25% glucose-based standard peritoneal dialysis solution. The degree of peritoneal pathological damage was assessed by Hematoxylin and eosin (H&E) staining. Peritoneal collagen deposition was detected by Masson's trichrome staining. The levels of Interleukin-6 (IL-6), Tumor necrosis factor-α (TNF-α), Interleukin-1β (IL-1β) and monocyte chemoattractant protein-1 (MCP-1) in the serum were measured by Enzyme Linked Immunosorbent Assay (ELISA). The expression levels of TGF-β, α-smooth muscle actin (α-SMA) and collagen I were examined by western blotting and immunohistochemistry. The protein expression levels and mRNA levels of E-cadherin, N-cadherin, vimentin, janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) in peritoneal tissue were determined by western blotting and qRT-PCR. TGF-β1 stimulated human peritoneal mesothelial cells (HPMCs), and the cells were treated with allicin and the JAK2/STAT3 pathway activator colivelin alone or in combination. A cell counting kit-8 (CCK-8) assay was used to measure cell viability. The role of JAK2/STAT3 in the effects of allicin was confirmed via in vitro mechanistic research by western blotting, wound healing assays and Transwell assays. Allicin relieves the inflammatory response by downregulating the levels of IL-1β, IL-6, MCP-1 and TNF-α. Furthermore, allicin decreased the expression of TGF-β, α-SMA and collagen I. Allicin also alleviated epithelial-to-mesenchymal transition (EMT), as specifically manifested by increased E-cadherin and reduced N-cadherin and vimentin. Further studies revealed that allicin reduced the protein levels of JAK2, STAT3, p-JAK2, and p-STAT3. The results of the cellular experiments verified the above results. The ability of allicin to inhibit fibrosis and the EMT process was significantly attenuated after HPMCs were treated with colivelin. Taken together, these findings suggest that allicin inhibits inflammation and EMT, thereby improving PF, and this protective effect may be achieved by inhibiting the JAK2/STAT3 signaling pathway.

Outcome differences of emergency cesarean delivery in the delivery room versus the operating room: A study based on propensity score matching.

This study sought to compare the risks and outcomes associated with category I cesarean section procedures performed in the delivery room versus those performed in the operating room. The analysis included 126 singleton pregnant women who underwent inpatient delivery at the Second People's Hospital of Nanning between January 2021 and May 2024. Following propensity score matching, 21 cases were in the delivery room group, and 105 cases were in the operating room group. Parameters under investigation encompassed decision-to-delivery interval, incision-to-delivery interval, surgical duration, intraoperative blood loss, postoperative antibiotic duration, postoperative hospital stay length, postoperative fever incidence, adverse neonatal outcomes, and blood routine parameters. The decision-to-delivery interval was significantly shorter in the delivery room group than in the operating room group. Conversely, the delivery room group exhibited longer surgical durations, higher blood loss, prolonged postoperative antibiotic usage, extended hospital stays, and elevated white blood cell counts with statistical significance (p < 0.05). Nevertheless, no notable variations were observed between the groups in maternal and neonatal outcome indicators, such as adverse neonatal outcomes and postoperative fever rates. The outcomes suggest that the delivery room group showed increased risks compared with the operating room group, potentially indicating heightened vulnerabilities to bleeding and infection. Hence, it is advisable for patients to undergo surgery in the operating room unless the delivery room is equipped with sterile surgical facilities or in cases of urgent necessity.

Diagnostic value of ultrasound for community-acquired pneumonia in children and its correlation with serum PCT level and PCIS.

This study aimed to evaluate the diagnostic value of ultrasound for community-acquired pneumonia (CAP) in children. Clinical information of children diagnosed with CAP and a control group of healthy children was collected, and lung ultrasound detection was performed. The lung ultrasound score (LUS) was assessed, and venous blood samples were collected. Serum indexes, including white blood cell count, were analyzed using an automatic immunoassay analyzer, while serum procalcitonin (PCT) level was measured using an enzyme-linked immunosorbent assay. The pediatric critical illness score (PCIS) was also performed for all subjects. White blood cell count, absolute neutrophil count, and respiratory index were significantly higher in the CAP group than those in the control group, while the oxygenation index was markedly lower. Ultrasound detection results showed that the CAP group exhibited significantly higher detection rates of pleural effusion, interstitial lung changes, lung consolidation, B-lines, air bronchogram signs, and reduced or absent lung sliding signs compared with the control group. In addition, the LUS and PCT levels were markedly higher in the CAP group, while the PCIS was notably lower. Further analysis exhibited that the LUS in the CAP group was significantly positively correlated with PCT levels and negatively correlated with PCIS. The receiver operating characteristic curve indicated that the area under the curve of LUS for diagnosing children with lung infection was 0.841. LUS is closely related to serum PCT level and PCIS. Lung ultrasound detection demonstrates high sensitivity and specificity, indicating its valuable clinical diagnostic utility for CAP in children.

Transcription factor YY1-activated GNG5 facilitates glioblastoma cell growth, invasion, stemness and glycolysis through Wnt/β-catenin pathway

G protein subunit Gamma 5 (GNG5) has been found to be involved in regulating glioma progression. However, its function and mechanism in glioblastoma (GBM) progression need to be further elucidated. GBM cell proliferation, apoptosis, invasion and stemness were assessed by cell counting kit 8 assay, EdU assay, flow cytometry, transwell assay and sphere formation assay. The mRNA and protein levels of GNG5 and Yin Yang 1 (YY1) were determined by quantitative real-time PCR and western blot (WB). Detection of the glucose consumption, lactate production and ATP/ADP ratios were used to assess cell glycolysis. Besides, Wnt/β-catenin pathway-related protein levels were examined by WB. Mice xenograft model was also constructed to explore GNG5 roles in vivo. GNG5 was highly expressed in GBM, and its silencing inhibited GBM cell proliferation, invasion, stemness and glycolysis, while promoted apoptosis. Transcription factor YY1 could bind to the GNG5 promoter region and induce its expression. GNG5 overexpression reversed the inhibitory effects of YY1 silencing on GBM cell growth, invasion, stemness and glycolysis. YY1/GNG5 axis could activate the Wnt/β-catenin pathway, and Wnt/β-catenin pathway agonists SKL2001 could revert the effects of GNG5 silencing on GBM cell progression. Furthermore, GNG5 facilitated GBM tumor growth by mediating the Wnt/β-catenin pathway. YY1-mediated GNG5 promoted GBM progression through the Wnt/β-catenin pathway.

Advanced Bio-sensing Technologies for Sickle Cell Disease Diagnosis.

Sickle cell diseases are widespread in regions encompassing the Mediterranean, Middle East, sub-Saharan Africa, and specific parts of Asia, primarily due to the abnormal production of hemoglobin S. This genetic blood disorder stems from a mutation in the beta-globin gene, a crucial component of hemoglobin and the heme-containing protein found in red blood cells. Point mutations in the hemoglobin gene can be inherited as a heterozygous or homozygous pattern. These mutations disrupt the normal configuration of the protein, impeding its physiological function and altering the cell's shape, giving it a sickle-like appearance. The resulting sickle cells can lead to organ damage, intense physical discomfort, and anemia; in severe cases, the condition can be fatal. Early detection and effective treatment methods have the potential to progressively reduce the associated mortality rate over time. To diagnose sickle cell disease and its carrier states with unparalleled specificity, a variety of approaches have been developed. The most common method includes differential blood cell counts and their assessment, high-performance liquid chromatography (HPLC) and hemoglobin electrophoresis. Furthermore, innovative sensing technologies are currently under development, encompassing user-friendly, cost-effective and portable point-of-care devices that are capable of timely diagnosis at the genetic and molecular levels of these disorders. The review delves into a range of established and innovative strategies utilized in the detection of sickle cell disease, also underscoring the essential role played by diverse bio-sensing techniques in propelling the advancement of early diagnosis of SCD.

Research of Optical Properties and Biocompatibility in Different Zones of Multilayered Translucent Zirconia on Hydrothermal Aging

Objective: We assessed the changes in optical properties and biocompatibility of transition zones in multilayered translucent monolithic zirconia exposed to prolonged hydrothermal aging and compared the results to those with different yttrium oxide contents. Materials and Methods: Four types of zirconia blocks from IPS e.max ZirCAD were used: 3Y-TZP e.max ZirCAD LT (ZL), 4Y-TZP e.max ZirCAD MT (ZM), 5Y-TZP e.max ZirCAD MT Multi (ZT), and 3Y/5Y-TZP e.max ZirCAD Prime (ZP). A total of 120 specimens (15.0 mm diameter and 1.5 mm height) were fabricated and divided into three groups (n = 10). The aging process for the specimens was conducted in an autoclave set to 134 °C and 0.2 MPa, with durations of 0 h (control), 5 h (first aged), and 10 h (second aged). The optical properties and biocompatibility were analyzed, followed by a statistical analysis of the data (α = 0.05). Results: Before and after aging, ZL and ZP exhibited the lowest color changes. ZT exhibited the highest average transmittance and translucency parameter values, while ZL had the lowest. The water contact angle test showed the highest value in ZM and lowest in ZL across all the aging stages. ZL, ZM, and ZP showed a considerable decrease in the water contact angle; however, ZT did not. A cell counting kit-8 assay showed ZL had the highest value, while ZM had the lowest. A filamentous actin test exhibited the highest value in ZL and lowest in ZM. In the vinculin analysis, ZL and ZT exhibited the lowest values, whereas ZM and ZP had the highest. Conclusion: 3Y/5Y-TZP exhibited a balanced performance across critical parameters, such as color stability, translucency, and biocompatibility, aligning with 3Y-TZP. While 5Y-TZP demonstrated superior translucency, it confirmed the lowest color stability, whereas 3Y-TZP achieved the highest biocompatibility. These properties provide clinicians with a reliable material option that ensures superior esthetic outcomes and long-term prognosis, ultimately contributing to improved patient satisfaction and clinical longevity.

Diagnostic accuracy of bone marrow blood evaluation in haemophagocytic lymphohistiocytosis paediatric patients.

Haemophagocytic lymphohistiocytosis (HLH) is a rare and serious immunological syndrome that involves a strong activation of cytotoxic T lymphocytes and macrophages. HLH determines a cytokine-mediated tissue injury with a contemporary multi-organ failure and a high fatality rate. A retrospective study was performed considering the medical records of paediatric patients who underwent a bone marrow aspirate for suspect HLH. The biomarkers evaluated were among those included in the HLH-2004. Lactate dehydrogenase (LD) was also evaluated. Haemophagocytosis was evaluated in bone marrow blood smear slides. Enrolled were 11 patients included in the HLH group and 8 patients as controls. Haemoglobin and fibrinogen resulted lower in HLH patients than in controls, while blood triglycerides, serum ferritin and LD resulted increased. Blood triglycerides and fibrinogen discriminated HLH cases perfectly, with a sensitivity and specificity of 100%. Ferritin had a sensitivity of 100% and a specificity of 83% (cut off ≥3,721µg/L) and LD of 73% and of 100% (the cut off ≥1,903U/L). Haemoglobin was found to have a sensitivity of 75% and a specificity of 100% (cut off ≤ 96g/L). Total haemophagocytes cell counts were not different between patients and controls. Only the increased number of phagocytized nucleated red blood cells (NRBC) was found to be significantly increased in the patients. Erythrocytes phagocytosis (≥4/1,000 cells) only tended towards significance. The blood biomarkers showed better diagnostic performance than the morphological evaluation. Among the different cell lineages engulfed by haemophagocytes, the best diagnostic performance was obtained by phagocytosed mature erythrocytes and immature nucleated erythrocytes.

Identification of immune traits associated with neurodevelopmental disorders by two-sample Mendelian randomization analysis

BackgroundOne of the main causes of health-related issues in children is neurodevelopmental disorders (NDDs), which include attention-deficit hyperactivity disorder (ADHD), autism spectrum disorder (ASD), and Tourette syndrome (TS). Nonetheless, there is relatively little prior research looking at the link between immunological inflammation and NDDs. Our work uses a two-sample Mendelian Randomization (MR) approach to provide a thorough evaluation of the causal effects of immune traits on ADHD, ASD, and TS.MethodsAs exposures, 731 immunological traits’ genetic associations were chosen, and the outcomes were genome-wide association data for ADHD, ASD, and TS. The inverse-variance weighted (IVW), weighted median (WM), and MR-Egger methods were used to conduct MR analysis. The results’ robustness, heterogeneity, and horizontal pleiotropy were confirmed using extensive sensitivity analysis.ResultsWith single-nucleotide polymorphisms serving as instruments and false discovery rate (FDR) correction applied, the study found that significantly higher expression of CD62L on CD62L+ myeloid DC (IVW, OR: 0.926, 95% CI 0.896~0.958, P = 9.42 × 10–6, FDR = 0.007) and suggestively higher absolute cell count (AC) of CD28 + DN (CD4-CD8-) (IVW, OR: 0.852, 95% CI = 0.780 ∼ 0.932, P-value = 4.65 × 10−4, FDR = 0.170) was associated with a lower risk of ADHD. There was no pleiotropy, and the causal relationships were strong according to sensitivity, leave-one-out, and MR-Steiger directionality tests. For ASD and TS, no harmful or protective immune traits were observed.ConclusionsThe results of the study lend credence to the theory that deficiency in CD62L on CD62L+ myeloid DC and CD28 + DN (CD4-CD8) AC may contribute to the onset of ADHD.

Comparative evaluation of surface roughness and bacterial adhesion on two bioactive cements: an in-vitro study

BackgroundDental restorative materials are recognized as artificial niches that facilitate the adherence and accumulation of oral microorganisms. To mitigate oral diseases and extend the lifespan of restorations, it is advantageous to use dental materials that exhibit low susceptibility to bacterial adhesion.ObjectiveTo evaluate and compare bacterial adhesion on two bioactive restorative materials, a glass hybrid restorative, and an alkasite with a nanohybrid resin composite as a positive control. The secondary objectives were to compare the surface roughness (SR) of the materials and determine the correlation between the bacterial adhesion and the SR.Materials and methodsThe samples consisted of 33 polished discs of each material: Group A: Tetric® N-Ceram (nanohybrid resin composite), Group B: Equia Forte™ HT Fil (glass hybrid restorative) and Group C: Cention N® (alkasite). Streptococcus mutans cultures were inoculated and after 24-hours of incubation, bacterial adhesion was measured by measuring optical density (OD) and number of colony forming units (CFUs). After 96-hours incubation, the bacterial cell count was determined using scanning electron microscopy (SEM). SR was assessed using surface profilometer.ResultsAlkasite had significantly lower OD and CFUs (p < 0.001 and p = 0.015 respectively). According to the SEM analysis, the glass hybrid restorative had lower mean bacterial cell count with no significant difference between the groups. The nanohybrid composite had the smoothest surface that was significantly lower than the alkasite and glass hybrid restorative (p = 0.002). None of the groups demonstrated a correlation between bacterial adhesion and SR.ConclusionAlkasite impedes bacterial adhesion better than the glass hybrid restorative and nanohybrid composite, while smoother surfaces are achieved with the nanohybrid composite.

Optimization of Fermentation and Biocontrol Efficacy of Bacillus atrophaeus XHG-1-3m2

Biological control plays an increasingly important role in various aspects of modern agriculture and forestry. Identifying biocontrol strains with commercial potential for effective disease management is currently a focal point in biological control research. In this study, Bacillus atrophaeus XHG-1-3m2, a strain with significant biocontrol potential against Wilsonomyces carpophilus causing shot hole disease in wild apricots, was developed. The study determined the antibacterial activity of the fermentation broth, the optimal fermentation medium composition and conditions, and explored its effectiveness in controlling Wilsonomyces carpophilus. The optimal fermentation medium for strain XHG-1-3m2 comprises 12.5 g/L yeast extract, 12.5 g/L soy peptone, 10.0 g/L sodium chloride, 1 g/L ammonium chloride, 1 g/L potassium dihydrogen phosphate, 1 g/L disodium hydrogen phosphate, and 0.5 g/L magnesium sulfate heptahydrate. With an initial pH of 7.0, a liquid volume of 40%, an inoculum volume of 3%, and shaking incubation at 28 °C for 24 h, the viable cell count reached 14 × 109 CFU/mL. In vitro and in vivo tests on leaves revealed that the fermentation broth and the biocontrol biofertilizer derived from this strain inhibited the leaf lesions caused by Wilsonomyces carpophilus on wild apricots, achieving inhibition rates of 94.62% and 82.46%, respectively.