Reduction In Cell Count Research Articles

Enhanced pathogenicity and synergistic effects of co-infection with bovine viral diarrhea virus 1 and HoBi-like virus in cattle and guinea pigs

IntroductionThe Bovine Viral Diarrhea Virus 1 (BVDV1) and HoBi-like virus (BVDV3), both within the same genus, share genomic homology and exhibit low antigenic cross-reactivity despite presenting similar clinical manifestations. In 2021, a bovine respiratory disease complex (BRDC) outbreak on two cattle farms in the Inner Mongolia Autonomous Region of China resulted in ten fatalities.MethodsMetagenomic and metatranscriptomic analyses were used to identify viral agents, including a co-infection case. A genetic evolution analysis assessed the relationships with related strains. Experimental infections in guinea pigs and calves evaluated the pathogenicity of the virusesResultsPhylogenetic analysis of the BVDV3 isolate IM2201 revealed close relatedness to Brazilian strains, with 97.06% nucleotide homology to the highly virulent strain SV478/07. Experimental co-infection in guinea pigs resulted in more severe clinical signs, including fever, cough, diarrhea, and significant pathological changes, and led to a higher mortality rate (40%) compared to no mortality from single-virus infections with BVDV1 or BVDV3. Similarly, co-infected cattle exhibited more severe clinical signs, including bloody diarrhea and rectal temperatures exceeding 40°C, along with persistent viremia and nasal viral shedding from 7 to 21 days post-infection. Blood analysis revealed significant reductions in white blood cell counts, particularly in co-infected cattle.DiscussionThis study highlights the enhanced pathogenicity and synergistic effects of BVDV1 and BVDV3 co-infection, exacerbating disease severity.

Time-dependent corrosion behavior of EH36 steel caused by Pseudomonas aeruginosa based on big data monitoring technology

Marine microbial corrosion poses a significant threat to the safe service of marine engineering equipment. Previous studies have often failed to thoroughly analyze the continuous and prolonged microbial corrosion process, resulting in an incomplete understanding of microbial corrosion mechanisms involved at various stages and the development of ineffective control strategies. This study employed a corrosion big data online real-time monitoring technique to investigate the time-dependent corrosion behavior of EH36 steel caused by Pseudomonas aeruginosa in aerobic environments over a 30-d incubation period. It was found that P. aeruginosa accelerated the corrosion of EH36 steel in the early stages by enhancing the cathodic oxygen reduction process. As oxygen levels declined, P. aeruginosa transitioned from aerobic to anaerobic respiration, promoting corrosion through biocatalytic nitrate reduction. In the later stages, the reduction in sessile cell counts, extreme low oxygen concentration, and dense surface film increased the charge transfer and film resistances, ultimately leading to corrosion inhibition. The weight loss and electrochemical data confirmed the effectiveness of the big data monitoring technique in investigating microbial corrosion, which provides new approaches for diagnosing and preventing microbial corrosion.

Simultaneous treatment with tap water-based neutral electrolyzed water and citric acid for inactivating foodborne pathogens on stainless steel and their synergistic bactericidal mechanisms

Electrolyzed water has many potential applications, the primary one being the eliminating microorganisms in the food industry. However, this application is limited by high chlorine levels. Tap water-based neutral electrolyzed water (TNEW) has the advantage of relatively low chlorine concentration (1.7–4.6 mg/L), but its sterilization effect is limited. In this study, TNEW was combined with citric acid (CA) solution, resulting in strong synergistic inactivation effect on Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on stainless steel surfaces. Simultaneous treatment of stainless steel with TNEW [available chlorine concentration (ACC) of 4.97 mg/L] and 0.1% CA for 15 min resulted in 4.78-, 4.26-, and >5.69 log CFU/cm2 surface reduction in E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, which was a synergistic cell count reductions of 3.25, 2.83, and >3.84 log units, respectively. Fluorescence and RT-qPCR analyses identified synergistic production of intracellular reactive oxygen species (ROS), damage to the cell membrane resulting from lipid peroxidation, and DNA damage as the major factors contributing to the synergistic lethal effect. The results of this study suggest that the TNEW–CA combination treatment can be applied as a potential substitute for conventional electrolyzed water treatment for food contact surface disinfection.

Evaluation of inhibitory potential of the flavonoid-rich fraction of Myrica esculenta Buch.-Ham. ex D. Don against DSS-induced colonic inflammation in mice

ObjectiveMyrica esculenta (family Myricaceae), a valuable plant species in China and India, has been traditionally used by many tribes and local communities to manage gut disorders. Scientific validation of its anti-ulcerative colitis activity was aimed. MethodsThe ethyl acetate fraction of Myrica esculenta (MeEa) was prepared and evaluated for its potency against DSS-induced ulcerative colitis (UC) in mice at 200 and 400 mg/kg BW oral dose. The effective dose of MeEa was determined through its effect on DSS-induced UC and was further analyzed through its effects on disease activity index (DAI), colon length, colon weight/length ratio, spleen weight, serum and colon tissue cytokine level, cell count and hemoglobin content. Further, the effect was determined through histopathology and FITC-dextran-induced membrane permeability assay. ResultsBetween the two doses, MeEa at 400 mg/kg BW was found to be the most effective dose in terms of reduced DAI scores, protected colon length from shortening, decreased colon weight/length ratio, reduced spleen weight, decreased pro-inflammatory cytokine level and stabilized the anti-inflammatory cytokine level in serum and colon tissue. MeEa 400 reduced cell counts and increased hemoglobin content and platelet count. MeEa 400 also prevented the colon by protecting epithelial cells and crypts. MeEa 400 provided significant protection from intestinal leakage and reduced FITC dextran level in serum. DiscussionMeEa 400 possesses significant anti-inflammatory potential and acts via attenuation of DSS-induced UC and inhibition of DAI scores. It reduces pro-inflammatory cytokines and stabilizes anti-inflammatory cytokine levels, reduces cell count, and protects epithelial tissue and crypts in the colon, as well as intestinal membrane leakage that occurred due to FITC-dextran administration in mice.

A-140 Reducing cell count bias with the GloCyte® Automated Cell Counter for CSF: an observational study at Loyola University Medical Center

Abstract Background Accurate STAT cerebrospinal fluid (CSF) cell counts are critical in supporting diagnoses of meningitis, malignancy, demyelinating disease, and hemorrhage. Limitations of automated analyzers to achieve acceptable linearity for the clinically relevant range has maintained manual CSF cell counting as the preferred method. Manual cell counting, performed with a hemocytometer and bright field microscope, is time-consuming, requires specialized skills and depends on human interpretation. This subjectivity inherently introduces the potential for non-random error in the enumeration of both total nucleated cells (TNC) and red blood cells in CSF by manual methods. The purpose of this study was to evaluate and compare the level of bias inherent in manual determination (TNC) in CSF as compared to the GloCyte Automated Cell Counter. Methods Result bias data for CSF specimens processed in the hematology lab at Loyola University Medical Center, a quaternary care 547-bed academic medical center in Maywood, IL, was analyzed for manual cell counting using a Neubauer-improved hemocytometer and GloCyte, an FDA-cleared cell counting device employing fluorescence technology to automate RBC and TNC counts in under 5 minutes, with linearity down to 0 cells/µL. Result bias was determined by defining the “expected result” for both GloCyte and hemocytometer as the initial cell count determined at a 1:5 dilution of a freshly-collected CSF sample and calculating the expected cell count for subsequent serial dilutions of 1:10, 1:20, 1:50, 1:100 for each method. The actual TNC count and the disparity of the actual result from the expected result was recorded as bias. Results Data are summarized below in the Table, and indicate a lower bias for GloCyte compared to manual counts across the range of dilutions. Conclusions This study demonstrates that GloCyte can be implemented as a means of effectively mitigating the reporting bias of CSF TNC counts performed with a Neubauer-improved hemocytometer.

Astragalus polysaccharide ameliorates bone marrow haematopoietic failure in mice with aplastic anaemia via the Hippo/TERT signalling pathway

ObjectiveTo study the effects of astragalus polysaccharide (APS) on aplastic anaemia (AA) in mice through modulation of the upstream and downstream effectors of the Hippo pathway. Further, we investigated the underlying mechanisms of APS in the treatment of AA to provide possible therapeutic strategies and drugs for clinical use. MethodsBALB/c mice were subjected to 4 Gy X-ray irradiation and subsequently injected with lymphocytes from DBA/2 donor mice by using the tail vein to establish an AA mouse model. Next BALB/c mice were randomly divided into five groups: control (untreated), AA (model), and three APS remedy groups - LA (low-dose APS, 200 mg/kg), MA (medium-dose APS, 400 mg/kg), and HA (high-dose APS, 800 mg/kg). The mice were sacrificed after 14 d of continuous gavage with different doses of APS or saline, and the bone marrow, spleen and liver were obtained. The haematopoietic condition, apoptosis levels, and the expression of Large Tumour Suppressor 1/2 (LATS 1/2), transcriptional co-activator of activation (TCA) Yes-associated protein (YesAP), transcriptional enhancer of adherence domain (TEAD) protein, and Telomerase Reverse Transcriptase (TERT), were analysed. ResultsCompared with control mice, mice in the AA group exhibited a reduction in peripheral blood cell counts, a significant decrease in the haematopoietic area, increased adipocyte infiltration, and overall haematopoietic failure, mirroring the clinical presentation of AA; After 14 days of treatment with different doses of APS, the APS-treated group, in comparison to the untreated AA group, showed an increase in peripheral blood cell counts (P < 0.05), an expansionof the haematopoietic area in the bone marrow, restoration of haematopoietic function (P < 0.05), and a marked reduction in adipocyte infiltration within the bone marrow(P < 0.05). Cell infiltration into the bone marrow was also reduced. Further experiments revealed that the expression levels of TERT, LATS 1/2, YAP, and TEAD were elevated in the treated mice (P < 0.05). ConclusionX-ray irradiation combined with T-lymphocyte infusion successfully established an AA mouse model, and APS administration modulated the expression of TERT and the Hippo pathway effectors LATS 1/2, YAP, and TEAD, which are implicated in the pathogenesis of AA. This modulation resulted in the restoration of bone marrow haematopoietic function in AA mice, suggesting APS as a promising therapeutic agent for the treatment of AA.

Ablation of the Integrin CD11b Mac-1 Limits Deleterious Responses to Traumatic Spinal Cord Injury and Improves Functional Recovery in Mice.

Spinal cord injury (SCI) triggers microglial/monocytes activation with distinct pro-inflammatory or inflammation-resolving phenotypes, which potentiate tissue damage or facilitate functional repair, respectively. The major integrin Mac-1 (CD11b/CD18), a heterodimer consisting of CD11b and CD18 chains, is expressed in multiple immune cells of the myeloid lineage. Here, we examined the effects of CD11b gene ablation in neuroinflammation and functional outcomes after SCI. qPCR analysis of C57BL/6 female mice showed upregulation of CD11b mRNA starting from 1 d after injury, which persisted up to 28 d. CD11b knockout (KO) mice and their wildtype littermates were subjected to moderate SCI. At 1 d post-injury, qPCR showed increased expression of genes involved with inflammation-resolving processes in CD11b KO mice. Flow cytometry analysis of CD45intLy6C-CX3CR1+ microglia, CD45hiLy6C+Ly6G- monocytes, and CD45hiLy6C+Ly6G+ neutrophils revealed significantly reduced cell counts as well as reactive oxygen species (ROS) production in CD11b KO mice at d3 post-injury. Further examination with NanoString and RNA-seq showed upregulation of pro-inflammatory genes, but downregulation of the ROS pathway. Importantly, CD11b KO mice exhibited significantly improved locomotor function, reduced cutaneous mechanical/thermal hypersensitivity, and limited tissue damage at 8 weeks post-injury. Collectively, our data suggest an important role for CD11b in regulating tissue inflammation and functional outcome following SCI.

Utilizing diffusion tensor imaging as an image biomarker in exploring the therapeutic efficacy of forniceal deep brain stimulation in a mice model of Alzheimer’s disease

Objective.With prolonged life expectancy, the incidence of memory deficits, especially in Alzheimer's disease (AD), has increased. Although multiple treatments have been evaluated, no promising treatment has been found to date. Deep brain stimulation (DBS) of the fornix area was explored as a possible treatment because the fornix is intimately connected to memory-related areas that are vulnerable in AD; however, a proper imaging biomarker for assessing the therapeutic efficiency of forniceal DBS in AD has not been established.Approach.This study assessed the efficacy and safety of DBS by estimating the optimal intersection volume between the volume of tissue activated and the fornix. Utilizing a gold-electroplating process, the microelectrode's surface area on the neural probe was increased, enhancing charge transfer performance within potential water window limits. Bilateral fornix implantation was conducted in triple-transgenic AD mice (3 × Tg-AD) and wild-type mice (strain: B6129SF1/J), with forniceal DBS administered exclusively to 3 × Tg-AD mice in the DBS-on group. Behavioral tasks, diffusion tensor imaging (DTI), and immunohistochemistry (IHC) were performed in all mice to assess the therapeutic efficacy of forniceal DBS.Main results.The results illustrated that memory deficits and increased anxiety-like behavior in 3 × Tg-AD mice were rescued by forniceal DBS. Furthermore, forniceal DBS positively altered DTI indices, such as increasing fractional anisotropy (FA) and decreasing mean diffusivity (MD), together with reducing microglial cell and astrocyte counts, suggesting a potential causal relationship between revised FA/MD and reduced cell counts in the anterior cingulate cortex, hippocampus, fornix, amygdala, and entorhinal cortex of 3 × Tg-AD mice following forniceal DBS.Significance.The efficacy of forniceal DBS in AD can be indicated by alterations in DTI-based biomarkers reflecting the decreased activation of glial cells, suggesting reduced neural inflammation as evidenced by improvements in memory and anxiety-like behavior.

High-performance binary to gray code converter: balancing energy use and thermal stability

Quantum Dot Cellular Automata (QCA) is an advanced technology in quantum electronics, leveraging quantum cells as its fundamental unit. This article introduces a design for a Binary to Gray (BG) code converter using the QCA technology. The proposed design uses fewer cells than previous models and extends the bit size capability to five bits in a single layer to minimize complexity and improve efficiency. The primary goal is to develop energy and thermal efficient BG code converters. These designs achieve a cell count reduction of 45.16% for two-bit, 29.54% for three-bit, and 25.45% for four-bit converters while improving the overall area by 41.17%, 29.54%, and 40% for two-bit,three-bit, and four-bit converters, respectively, with a latency of 0.5. The 55-cell, five-bit BG converter takes up 0.07μ m 2 and has a latency of 0.5. Comprehensive simulations were conducted using the QCADesigner, QCADesignerE 2.0.3, and QCA Pro tools to validate the proposed design’s functionality.

3D Melt-Extrusion Printing of Medium Chain Length Polyhydroxyalkanoates and Their Application as Antibiotic-Free Antibacterial Scaffolds for Bone Regeneration.

In this work, we investigated, for the first time, the possibility of developing scaffolds for bone tissue engineering through three-dimensional (3D) melt-extrusion printing of medium chain length polyhydroxyalkanoate (mcl-PHA) (i.e., poly(3-hydroxyoctanoate-co-hydroxydecanoate-co-hydroxydodecanoate), P(3HO-co-3HD-co-3HDD)). The process parameters were successfully optimized to produce well-defined and reproducible 3D P(3HO-co-3HD-co-3HDD) scaffolds, showing high cell viability (100%) toward both undifferentiated and differentiated MC3T3-E1 cells. To introduce antibacterial features in the developed scaffolds, two strategies were investigated. For the first strategy, P(3HO-co-3HD-co-3HDD) was combined with PHAs containing thioester groups in their side chains (i.e., PHACOS), inherently antibacterial PHAs. The 3D blend scaffolds were able to induce a 70% reduction of Staphylococcus aureus 6538P cells by direct contact testing, confirming their antibacterial properties. Additionally, the scaffolds were able to support the growth of MC3T3-E1 cells, showing the potential for bone regeneration. For the second strategy, composite materials were produced by the combination of P(3HO-co-3HD-co-HDD) with a novel antibacterial hydroxyapatite doped with selenium and strontium ions (Se-Sr-HA). The composite material with 10 wt % Se-Sr-HA as a filler showed high antibacterial activity against both Gram-positive (S. aureus 6538P) and Gram-negative bacteria (Escherichia coli 8739), through a dual mechanism: by direct contact (inducing 80% reduction of both bacterial strains) and through the release of active ions (leading to a 54% bacterial cell count reduction for S. aureus 6538P and 30% for E. coli 8739 after 24 h). Moreover, the composite scaffolds showed high viability of MC3T3-E1 cells through both indirect and direct testing, showing promising results for their application in bone tissue engineering.

An approach to combat multidrug-resistant K. pneumoniae strain using synergistic effects of Ocotea diospyrifolia essential oil in combination with amikacin

The natural antimicrobial properties of essential oils (EOs) have contributed to the battle against multidrug-resistant microorganisms by providing new ways to develop more effective antibiotic agents. In this study, we investigated the chemical composition of Ocotea diospyrifolia essential oil (OdOE) and its antimicrobial properties combined with amikacin (AMK). Through gas chromatography-mass spectrometry (GCMS) analysis, the primary constituents of OdOE were identified as α-bisabolol (45.8 %), β-bisabolene (9.4 %), γ-elemene (7.6 %), (Z)- β-farnesene (5.2 %), spathulenol (3.5 %), (Z)-caryophyllene (3.3 %), and (E)-caryophyllene (3.1 %). In vitro assessments showed that the combined administration of OdOE and AMK exerted a synergistic antibacterial effect on the multidrug-resistant K. pneumoniae strain. This synergistic effect demonstrated bacteriostatic action. OdEO combined with amikacin showed protein extravasation within 2 h of treatment, leading to bacterial death, which was determined by a reduction in viable cell count. The effective concentrations showed hemocompatibility. In vivo assessments using Caenorhabditis elegans as a model showed the survival of 85 % of infected nematodes. Therefore, the combination OdEO combined with amikacin exhibited antimicrobial activity against a multidrug-resistant K. pneumoniae strain. Thus, OdOE is a promising agent that may be considered for development of antimicrobial treatment.

DDX41 haploinsufficiency causes inefficient hematopoiesis under stress and cooperates with p53 mutations to cause hematologic malignancy

Germline heterozygous mutations in DDX41 predispose individuals to hematologic malignancies in adulthood. Most of these DDX41 mutations result in a truncated protein, leading to loss of protein function. To investigate the impact of these mutations on hematopoiesis, we generated mice with hematopoietic-specific knockout of one Ddx41 allele. Under normal steady-state conditions, there was minimal effect on lifelong hematopoiesis, resulting in a mild yet persistent reduction in red blood cell counts. However, stress induced by transplantation of the Ddx41+/− BM resulted in hematopoietic stem/progenitor cell (HSPC) defects and onset of hematopoietic failure upon aging. Transcriptomic analysis of HSPC subsets from the transplanted BM revealed activation of cellular stress responses, including upregulation of p53 target genes in erythroid progenitors. To understand how the loss of p53 affects the phenotype of Ddx41+/− HSPCs, we generated mice with combined Ddx41 and Trp53 heterozygous deletions. The reduction in p53 expression rescued the fitness defects in HSPC caused by Ddx41 heterozygosity. However, the combined Ddx41 and Trp53 mutant mice were prone to developing hematologic malignancies that resemble human myelodysplastic syndrome and acute myeloid leukemia. In conclusion, DDX41 heterozygosity causes dysregulation of the response to hematopoietic stress, which increases the risk of transformation with a p53 mutation.

Fig seed oil improves intestinal damage caused by 5‐FU‐induced mucositis in rats

AbstractIntestinal mucositis poses a significant concern associated with cancer therapy. This study aims to investigate the protective and/or healing effect of fig seed oil (FSO) on 5‐fluorouracil (5‐FU)‐induced intestinal mucositis by targeting inflammatory markers and histologic changes in rats. Albino Wistar adult rats were randomly divided into four groups, including three male and three female animals. All the animals in the four groups had a normal standard diet and water throughout the experimental period, which lasted up to 11 days. Rats were administered FSO 0.6 mL (mucositis FSO group) and FSO 0.2 mL (mucositis FSO‐R group) daily throughout the experiment. These two groups and one additional group (mucositis group) were given an intraperitoneal injection of 5‐FU (300 mg/kg) on Day 5 of the experiment. In contrast, the fourth group (Control group) was given an intraperitoneal saline injection on Day 5 of the experiment. FSO treatment ameliorated 5‐FU‐induced intestinal mucositis. On immunohistologic examination, FSO suppressed significantly the activation of NF‐κB and expression of IL‐β and TNF‐α of the harvested intestinal tissue. The reduced dose FSO (mucositis FSO‐R) was as effective as the full dose (mucositis FSO) in suppressing IL‐β and TNF‐α production, but was not as effective as the full dose in suppressing NF‐κB. On light microscopy, FSO attenuated significantly 5‐FU‐induced anomalies, such as the reduction of intestinal villus length and Goblet cell count. The reduced dose FSO (mucositis FSO‐R) was as effective as the full dose (mucositis FSO) in restoring villus length, but was not as effective as the full dose in restoring Goblet cell count. The findings of the study suggest that FSO inhibits 5‐FU‐induced intestinal mucositis via modulation of mucosal inflammation.

Design and simulation of a new QCA-based low-power universal gate

Quantum-dot Cellular Automata (QCA) is recognized in electronics for its low power consumption and high-density capabilities, emerging as a potential substitute for CMOS technology. GDI (Gate Diffusion Input) technology is featured as an innovative approach for enhancing power efficiency and spatial optimization in digital circuits. This study introduces an advanced four-input Improved Gate Diffusion Input (IGDI) design specifically for QCA technology as a universal gate. A key feature of the proposed 10-cell block is the absence of cross-wiring, which significantly enhances the circuit’s operational efficiency. Its universal cell nature allows for the carrying out of various logical gates by merely altering input values, without necessitating any structural redesign. The proposed design showcases notable advancements over prior models, including a reduced cell count by 17%, a 29% decrease in total energy usage, and a 44% reduction in average energy loss. This innovative IGDI design efficiently executes 21 combinational and various sequential functions. Simulations in 18 nm technology, accompanied by energy consumption analyses, demonstrate this design’s superior performance compared to existing models in key areas such as multiplexers, comparators, and memory circuits, alongside a significant reduction in cell count.

Tucidinostat plus toripalimab in patients with unresectable or metastatic melanoma: An open-label, single-arm phase II study.

e21527 Background: Acral and mucosal melanoma respond poorly to anti–PD-1 monotherapy. Tucidinostat, a benzamide class of histone deacetylase (HDAC) inhibitor, potentially augments immunotherapy with PD-1 blockade. We therefore conducted a phase II study to investigate the safety and efficacy of tucidinostat plus toripalimab in unresectable or metastatic melanoma. Methods: This was a single center, single arm phase II clinical trial. Patients with pathologically confirmed locally advanced unresectable or metastatic melanoma were enrolled and received 30 mg tucidinostat orally twice a week and 240 mg toripalimab every 3 weeks until disease progression or unacceptable toxicity. The primary endpoint was the confirmed ORR per RECIST v1.1. Secondary endpoints included progression free survival (PFS), disease control rate (DCR), overall survival (OS), and safety. Results: Between October 2022 and May 2023, a total of 21 patients(pts) were enrolled. All pts were included in the safety analysis set and 18 pts were included in the full analysis set (FAS) for efficacy analysis. In the FAS, the median age was 56.0 years (range: 49 to 60), 6 pts (33.3%) were acral melanoma, 9 (50.0%) mucosal melanoma, 1 (5.6%) cutaneous melanoma and 2 (11.1%) unknown primary melanoma. 15 pts were treated as first-line, and 3 pts were as second-line. By Dec 2023, the median follow-up was 8.8 mo (95% CI:7.6-10.4) , Confirmed ORR was 27.8% (95% CI:9.7-53.5%). The DCR was 61.1% (95% CI:35.7-82.7%). Median progression-free survival was 4.1 months (95%CI: 1.4-NR) ,OS was not reached. Subgroup analysis showed that, in acral and mucosal melanoma pts, the confirmed ORR was 50.0%(95%CI:11.8-88.2%) and 22.2%(95%CI: 2.8-60.0%), respectively. The incidence of treatment-related adverse events (TRAEs) was 90.5% (19/21). The most common ≥3 grade TRAEs were neutrophil reduction (28.6%), white blood cell count reduction (9.5%), alanine aminotransferase increase (9.5%), platelet count reduction (9.5%), aspartate aminotransferase increase (9.5%), and γ-glutamyl transferase increase (9.5%). No treatment-related deaths occurred. Conclusions: The combination of tucidinostat and toripalimab showed acceptable safety and encouraging efficacy in patients with unresectable or metastatic melanoma. Further research is warranted. Clinical trial information: NCT05478473 .

The use of organic peroxyacids for the inactivation of calcium-mediated biofilm formation by Bacillus licheniformis

Biofilms formed by Bacillus licheniformis usually serve as a continuous bacterial contamination in the dairy industry. The purpose of this study was to explore how 1% organic acids (citric acid, phenyllactic acid, malic acid, and butyric acid) and their 1% organic peroxyacids (peroxycitric acid, peroxyphenyllactic acid, peroxymalic acid, and peroxybutyric acid) can inactive the planktonic and biofilm cells of B. licheniformis. Results demonstrated that the number of B. licheniformis planktonic cells was increased from 4 to 6.46 Log CFU mL−1 when treated with water. Organic peroxyacids, especially peroxyphenyllactic acid (PPA), obviously inactivated planktonic cells of B. licheniformis by a range of 3.32–4.14 Log CFU mL−1. Moreover, organic peroxyacids could also decrease the cell surface hydrophobicity of bacteria, and effectively control the Ca2+-mediated biofilm formation by B. licheniformis in 96-well plates (reduction of OD values by 0.14–0.33) and on stainless steel (reduction of cell counts by 1.20–2.49 Log CFU cm−2). The low minimum biofilm eradication concentration of PPA (0.125%) also suggests its strong biofilm inactivation activity, which could serve as a novel and environmentally friendly biofilm control agent in the dairy industry.

The effect of garlic formulation on some hematological parameters and organ weight of male and female Wister albino rats

The study was conducted to investigate the impact of garlic formulation on certain haematological parameters and organ weights in both male and female rats. Haematological data, including the count of red blood corpuscles (RBC), the volume of packed cells (PCV), the count of white blood corpuscles (WBC), and the count of platelets, were evaluated using a fully automated haematology analyser. The liver, right kidney, left kidney, heart, and spleen of the rats were weighed to determine their respective organ weights. A total of thirty (30) albino Wister rats, consisting of both males and females, were randomly assigned to five groups, with six animals in each group. Group 1 acted as the positive control and was given 100% standard feed. Group II rats were given a mixture of 10% garlic and 90% feed. Group III animals were provided with a diet consisting of 20% garlic and 80% feed. Group IV received a diet containing 40% garlic and 60% feed. The final group, Group V, was fed a diet consisting of 80% garlic and 20% feed daily for a period of 21 days. The feed formulation was assessed at concentrations of 10, 20, 40, and 80 g/dl, based on the weight of the animal. The findings indicated a noteworthy (P&lt;0.05) reduction in white blood cell count (WBC) and lymphocyte count (LYM). There were no statistically significant changes (P&gt;0.05) observed in the levels of haemoglobin (HGB), haematocrit (HCT), mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (MCV), and mean corpuscular haemoglobin (MCH) when compared to the control group. Furthermore, there was a noteworthy (P&lt;0.05) reduction in HGB, MCV, and MCH when the rats reached a body weight of 80 g/100dl. The results indicated a statistically significant (P&lt;0.05) reduction in the weight of the left kidney, heart, and spleen, as well as in the liver and right kidney weights at 20, 40, and 80 g/dl body weight. There were no statistically significant differences (P&gt;0.05) in the body weight of the liver and right kidney, which were both 10g/dl. Ultimately, the use of garlic formulation in appropriate dosages can impact certain haematological markers and the weights of organs.

#2084 Biologic effect evaluation of de novo long-term release tacrolimus on metabolizer phenotype and T cell composition after kidney transplantation

Abstract Background and Aims Kidney transplantation (KTX) is the current treatment of choice in patients with kidney failure. Since immunosuppressive medication to prevent acute rejection is still needed in high doses with frequent adverse events, new strategies to achieve risk-appropriate personalization of immunosuppression are needed. Calcineurin inhibitors are the cornerstone of T cell suppression, currently available in different release formulations including once daily long-term release tacrolimus (LCPT) and regular twice daily immediate release tacrolimus (IRT). Little is known about the temporal evolution of tacrolimus metabolism potentially influencing T cell composition and clinical endpoints between different tacrolimus formulations after KTX. Method To evaluate the influence of de novo LCPT on clinical endpoints, metabolizer phenotype and immune cell composition after KTX, subgroup analysis of a single-center, prospective, observational cohort study including 32 patients started on LCPT (0.1 mg/kg) and 55 patients started on IRT (0.1 mg/kg) was conducted. Peripheral blood mononuclear cells were isolated from whole blood samples drawn before KTX, 2 months and 12 months after KTX for flow cytometry analysis. Study visits were simultaneously conducted to obtain tacrolimus dosing, plasma levels, acute rejection and infection incidence. Statistical analysis was done using GraphPad Prism (v10), accepting significance for p &amp;lt; 0.05, and flow cytometry analysis was done using FlowJo (v10) software. Results The cohort represented KTX recipients with a median age of 60 years and similar distributions of gender, HLA-mismatches and induction therapy. Clinical outcome data showed a trend towards lower acute rejection rate, lower serum-creatinine values and increased incidence of BK-viremia in LCPT treated patients. Rejection rate was associated with a fast-metabolizer phenotype (HR 3.9; 1.1-6.8, p &amp;lt; 0.01) and HLA mismatch &amp;gt; 3 (HR 4.0; 1.2-7.3, p &amp;lt; 0.01) while this effect was blunted in the LCPT group (HR 1.9; 0.7-3.2, p = 0.67 and HR 0.9; 0.3-1.4, p = 0.38 respectively) in a multivariate Cox-regression model. Cluster exploration and tSNE analysis of flow cytometry data revealed diminution of activated-proliferative effector and regulatory T cell trajectories, with significant reduction in cell counts of CD4+CD15s+/CD45RA-Ki67+Foxp3+ Treg and in CD4+CD147high/CD45RA-LAP+HLADR+FcRL3+ effector T cells in LCPT treated patients after KTX. Conclusion Induction with de novo LCPT was associated with a blunted influence of metabolizer phenotype and HLA-mismatch on acute rejection in our cohort. Major changes were detected in peripheral T cell composition after transplantation, with signs of a stronger immunosuppressive effect of LCPT on circulating activated T cells. Therefore, LCPT may be considered for KTX recipients with higher HLA-mismatch (i.e. “old-for-old”) or fast-metabolizer phenotype. Inclusion of changes in T cell composition as biologic effect measures may be worthwhile in future studies evaluating biomarkers to guide immunosuppression and predict complications after KTX.

Management of hyperleukocytosis in pediatric acute myeloid leukemia using immediate chemotherapy without leukapheresis: results from the NOPHO-DBH AML 2012 protocol.

Hyperleukocytosis in pediatric acute myeloid leukemia (AML) is associated with severe complications and an inferior outcome. We report results on patients with hyperleukocytosis included in the NOPHO-DBH AML 2012 study. We recommended immediate initiation of full-dose chemotherapy (etoposide monotherapy for 5 days as part of the first course), avoiding leukapheresis and prephase chemotherapy. Of 714 patients included in the NOPHO-DBH AML 2012 study, 122 (17.1%) had hyperleukocytosis, and 111 were treated according to the recommendations with etoposide upfront without preceding leukapheresis or prephase chemotherapy. The first dose was applied the same day as the AML diagnosis or the day after in 94%. Etoposide was administered via peripheral veins in 37% of patients without major complications. After initiation of etoposide the white blood cell counts on days 2-5 were 69%, 36%, 17% and 8%, respectively, of the pre-treatment level. On day 3, 81% of patients had a white blood cell count <100 x109/L. Five-year event-free and overall survival rates for all patients with hyperleukocytosis were 52.9% (95% confidence interval [95% CI]: 44.4-63.0) and 74.1% (95% CI: 66.4-82.6), compared to 64.9% (95% CI: 60.9-69.1) and 78.9% (95% CI: 75.4-82.4) for patients without hyperleukocytosis (P<0.001 for event-free survival, P=0.1 overall survival). Six-week early mortality was 4.1% for all patients with hyperleukocytosis (2.7% for the 111 patients treated with etoposide upfront). We conclude that management of hyperleukocytosis in pediatric AML with immediate etoposide monotherapy without leukapheresis or prephase chemotherapy is feasible, safe and effective. The reduction in white blood cell count during the first days is comparable to the reported results of leukapheresis, and outcomes seem at least equivalent to therapies including leukapheresis. Based on our results, we advocate abandoning leukapheresis for hyperleukocytosis in pediatric AML. Instead, it is crucial to start induction chemotherapy as early as possible.

Biocontrol characteristics and application of phage SEP4 against multidrug-resistant Salmonella biofilm on food matrix

The emergence of antibiotic resistance and biofilm formation among Salmonella strains, especially in chicken meat, poses substantial risks to economic and public health. To mitigate these challenges, we explored interventions and biofilm inhibition strategies against multidrug-resistant serovars (B, C1, and D groups), by Salmonella phage SEP4, an isolate with broad host-spectrum. The morphological and genomic analyses classified it as a member of the Siphoviridae family with an icosahedral head of 124.56 ± 8.9 nm diameter and 66.28 ± 2.01 nm tail length, respectively. Phage SEP4 displayed robust survivability over a pH range of 3–10 and temperatures from 30 °C to 60 °C. Phage adsorption peaked at 73.20% in 15 min, with a burst size of 119 PFU/CFU at optimal MOI-1. Moreover, a linear double-stranded DNA genome of 41,828 bp, characterized by 49.84% GC content and putative 57 open reading frames (ORFs), and lacked antibiotic or virulence factor genes. Anti-Salmonella phage efficacy demonstrated a significant reduction in viable cell counts at 25 °C compared to 4 °C against tested MDR strains in food models. The bactericidal activity of phage SEP4 ranged from 14% to 46% and 25%–33.8% against host at MOI-1 and 100, while 22%–37% for other MDR strains. Simultaneously, SEP4 inhibited the growth of matured biofilms on lettuce leaves and chicken surfaces, achieving complete reduction within 4–6 h. SEM images confirmed the drastic destruction of cohesive biofilm architectures by phage treatment. The findings show that phage SEP4 has promising potential as a biocontrol agent, ensuring food safety by controlling MDR Salmonella contaminations and associated biofilms.