Fresh Blood Research Articles

A Scoping Review of Military Combat Casualty Data on Submassive, Massive, and Supermassive Transfusions.

Blood transfusions are common during combat casualty care, aiming to address the loss of blood volume that often accompanies severe battlefield injuries. This scoping review delves into the existing military combat casualty data to analyze the efficacy, challenges, and advances in the use of massive and super-massive transfusions in the management of critically injured warfighters. We performed a scoping review of combat-related literature published between 2006 and 2023 pertaining to massive transfusions used during combat deployments. We utilized PubMed to identify relevant studies and utilized the PRISMA-ScR Checklist to conduct the review. We identified 53 studies that met the inclusion criteria with the majority being retrospective studies from registries used by the United States, British, French, and Dutch Militaries. Most of the studies focused on transfusion ratios, the movement of blood transfusions to more forward locations, implementation of massive transfusions with different fibrinogen-to-red blood cell ratios, the addition of recombinant factor VII, and the use of predictive models for transfusion. Lastly, we identified reports of improved survival for casualties with the rapid implementation of various blood products (warm fresh whole blood, cold-stored low titer group O blood, freeze-dried plasma, and component therapy) and literature relating to pediatric casualties and submassive transfusions. Notable findings include the establishment of hemodynamic and cell blood count parameters as predictors of the requirement for massive transfusions and the association of higher fibrinogen-to-red blood cell ratios with decreased mortality. We identified 53 studies focused on blood transfusions from the Global War on Terrorism conflicts. The majority were related to transfusion ratios and the movement of blood transfusions to more forward locations. We highlight key lessons learned on the battlefield that have been translated into scientific developments and changes in civilian trauma methods.

Clinical and epidemiological features of imported loiasis in Beijing: a report from patients returned from Africa

BackgroundLoiasis is one of the significant filarial diseases for people living in West and Central Africa with wide endemic area but is not seen in China. As economy booms and international traveling increase, China faces more and more imported parasitic diseases that are not endemic locally. Loiasis is one of the parasitic diseases that enter China by travelers infected in Africa. The better understanding of the clinical and laboratory features of loa loa infection will facilitate the diagnosis and treatment of loiasis in China.MethodsThe study targeted travelers who were infected with L. loa in endemic Africa regions and returned to Beijing between 2014 and 2023. Epidemiological, clinical, and biological data as well as treatment of these patients were collected.ResultsTotal 21 cases were identified as L. loa infection based on their typical clinical manifestations and parasite finding. All cases had a history of travel to Africa for more than 6 months, most of them are the construction workers dispatched to West Africa with outdoor activities. Calabar swelling (n = 19; 90.5%) and pruritus (n = 11; 52.4%) were among the most common clinical symptoms followed by muscle pain (n = 7; 33.3%) and skin rash (n = 2; 9.5%). The adult worms were observed in the eyelid or subconjunctiva (n = 2; 9.5%) and subcutaneous tissues (n = 2; 9.5%). Although all patients presented with a high eosinophil count (> 0.52 × 109/L), only two cases displayed microfilariae in fresh venous blood and positive for filarial antigen. A cut section of adult worm was observed through biopsy on a skin nodule surrounded by lymphocytes, plasma cells and eosinophils. All subjects were positive in PCR targeting L. loa ITS-1. The constructed phylogenetic tree based on the amplified ITS-1 sequences identified their genetical relation to the L. Loa from Africa. All patients treated with albendazole and diethylcarbamazine were recovered without relapse.ConclusionThis study provides useful information and guideline for physicians and researchers in non-endemic countries to diagnose and treat loiasis and L. loa infections acquired from endemic regions.

Assessment of ID family proteins expression in colorectal cancer of Iraqi patients.

Colorectal cancer (CRC) is the second most deathly worldwide and third most common cancer, CRC is a very heterogeneous disease where tumors can form by both environmental and genetic risk factors and includes epigenetic and genetic alternations. Inhibitors of DNA binding proteins (ID) are a class of helix-loop-helix transcription regulatory factors; these proteins are considered a family of four highly preserved transcriptional regulators (ID1-4), shown to play significant roles in many processes that are associated with tumor development. ID family plays as negatively dominant antagonists of other essential HLH proteins, concluding the creation of non-functional heterodimers and regulation of the transcription process. 120 Fresh tissue and blood samples Forty (40) samples of fresh tissue and blood were collected from patients diagnosed with CRC, twenty (20) samples were collected from a patient diagnosed as healthy. The (qRT-PCR) method is a sensitive technique for the quantifying of steady-state mRNA levels that used to evaluation the expression levels of ID (1-4) gene. The findings indicate downregulation in ID1 in tissue with a highly significant change between patients and control groups, where upregulation in the ID1 gene is shown in blood samples.ID2 gene also demonstrated high significant change where show upregulation in tissue and downregulation in blood sample. ID3 and ID4 genes show downregulation in tissue and blood samples with a significant change in ID3 blood samples between patient and blood groups. Because of the regulation function of the ID family in many processes, the up or down regulation of IDs genes in tumors Proves how important its tumor development, and therefore those proteins can be used as an indicator for CRC.

Unlocking Healing Potential: A Case Study on the Management of Diabetic Foot Ulcer

Diabetes stands as one of the most prevalent diseases globally, and diabetic patients are at risk of developing foot ulcers as a microvascular complication. These ulcers stem from diabetic neuropathy or atherosclerosis, leading to ischemia or breakdown of glucose-laden tissue, resulting in inflammation and ulceration. If left untreated, these ulcers can become infected, spreading to the underlying tissues. In Ayurveda, such ulcers are termed as Madhumehajanya Vrana. Sushruta Acharya outlined Anushastra Karma for managing Vrana, which includes Ksharakarma (application of alkalis) and Jalukavacharana (leech therapy). Kshara functions by actions like Chhedana, Bhedana, Lekhana and Ropana. Jalukavacharana possesses anti-inflammatory properties, promotes fresh blood supply by removing vitiated Dosha from the blood, and fosters wound healing by forming healthy granulation tissue. A 65-year-old female patient presented with a non-healing wound on the plantar aspect of her left foot below the greater toe since 8 months, associated with foul-smelling pus discharge for 6 months. She had a known history of diabetes mellitus since 10 years. The ulcer measured 4cm x 3cm x 2.5cm, GRBS was 380mg/dl with HbA1c of 12%. Patient was treated with Ksharataila infiltration, application of Tilwaka pratisaraneeya kshara followed by Lekhana of callosity present over the edges of the ulcer, and Jalukavacharana therapy for four sessions with seven days interval, in addition to internal medications. Glycemic control was achieved by Inj. Insulin Mixtard 30/70 30 units in morning and 15units at night.

Evaluating the combined and individual cytotoxic effect of beauvericin, enniatin B and ochratoxin a on breast cancer cells, leukemia cells, and fresh peripheral blood mononuclear cells

Beauvericin (BEA), Enniatin B (ENN B), and Ochratoxin A (OTA) are mycotoxins produced by fungi species. Their main effect on several organs and systems is associated with chronic exposure going from immunotoxicity, estrogenic disorders, and renal failure to cancer (in animals and humans). OTA belongs to Group 1 according to the International Agency for Research in Cancer (IARC) and it has legislated limited values; not happening for BEA nor ENN B. Exposure to mixtures of mycotoxins occurs through food intake in daily consumption. The aim of this study was to evaluate the implication of BEA, ENN B, and OTA individually and combined in producing cytotoxicity in cells for immunological studies and cancer cell lines (human leukemia cells (HL-60), fresh human peripheral blood mononuclear cells (PBMCs), and human breast cancer (MDA-MB-231) cells). Cells were treated for 4 h and 24 h at different concentrations of BEA, ENN B, and OTA, respectively. Viability assays were carried out by flow cytometry using DAPI (4′,6-diamindino-2-phenylindole, dihydrochloride) as a viability dye and the potential effects of synergism, addition, and antagonism were assessed through the Chou and Talalay method. Individual OTA treatment exerted the greatest cytotoxicity for PBMC cells (IC50 0.5 μM) while ENN B for HL-60 (IC50 0.25 μM) and MDA-MB-231 (IC50 0.15 μM). In binary combination [ENN B + OTA] resulted in exerting the greatest cytotoxicity for HL-60 and MDA-MB-231 cells; while [BEA + OTA] in PBMC cells. The triple combination resulted in being highly cytotoxic for PBMC cells compared to HL-60 and MDA-MB-231 cells. In summary, PBMC cells were the most sensible cells for all three mycotoxins and the presence of OTA in any of the combinations had the greatest toxicity causing synergism as the most common cytotoxic effect.

Modifications and Optimization of an Autologous Intracerebral Hemorrhage Rat Model

Objective: To date, no novel drug therapies have proven effective in improving outcomes after intracerebral hemorrhage (ICH). This has been slowed, in part, by a lack of reliable preclinical models that recapitulate the pathophysiological consequences of ICH. Limited descriptions of the methodology of preclinical models may play a role in this failed translation. The current study aimed to improve the autologous blood injection model of ICH and provide a comprehensive methodology to facilitate a reliable preclinical model that bridges the translational research gap. Methods: A modified single-injection protocol for an autologous-blood ICH model was developed and tested. This rodent ICH model produces injury in the basal ganglia of adult rats via a stereotaxically-assisted right-sided single-injection of fresh autologous whole blood. A comprehensive step-wise description of the methodology is presented and documented in a written and visual format with supporting pictorialmaterial, to prevent common mistakes that result in failed hematoma formation.Rates of successful hematoma formation using the single-injection protocol, outlined here, were compared to those using the traditional double-injection protocol.

Cytotoxic effect of 1% and 3% Sodium hypochlorite as irrigation agent in primary teeth on human red blood cells: an in vitro study

Background and objectives. The endodontic treatment of primary teeth presents significant challenges due to their unique anatomical characteristics. To ensure a successful treatment, it is essential to evaluate a few factors. These factors include accurate diagnosis, disinfection protocol, intracanal medicaments, and irrigation solutions. While Sodium hypochlorite (NaOCl) has gained success as an irrigation agent in primary teeth, it is important to note that higher concentrations of NaOCl can potentially cause toxicity to the periapical environment if they pass through the tooth's apical foramina. Thus, this study compared the cytotoxicity of two different concentrations of sodium hypochlorite at varied volumes on red blood cells (RBC). Materials and methods. In order to evaluate the cytotoxicity potential, fresh human blood was collected from a single healthy individual and subjected to testing. For the test, 10 µL, 20 µL, 30 µL, 40 µL and 50 µL of 1% and 3% NaOCl were added to the RBC. The tubes were gently mixed and incubated at 37°C for 30 minutes. A UV-visible spectrophotometer was used to calculate the absorbance of the supernatant at a wavelength of 540 nm. Results. There was no significant difference in 1% sodium hypochlorite at different volumes with a p value >0.05 using Tukey's post hoc test. However, there was a significant difference between the volumes of 3% sodium hypochlorite at 40 and 50 µg/ml with a p value of 0.004 and 0.001 respectively. Conclusions. According to the current study's findings, 1% sodium hypochlorite solution exhibited less toxicity than 3% sodium hypochlorite.

Adapting the SMART tube technology for flow cytometry in feline full blood samples.

Flow cytometry of blood samples is a very valuable clinical and research tool to monitor the immune response in human patients. Furthermore, it has been successfully applied in cats, such as for infections with feline immune deficiency virus (FIV). However, if cells are not isolated and frozen, analysis of anticoagulated blood samples requires mostly prompt processing following blood collection, making later analysis of stored full blood samples obtained in clinical studies often impossible. The SMART Tube system (SMART TUBE Inc., California, United States; SMT) allows fixation and long-term preservation of whole blood samples at -80°C. However, this system has so far only been applied to human biological samples. In the present study, a new flow cytometry SMART Tube protocol adapted for feline whole blood samples was successfully established allowing quantification of T-helper cells, cytotoxic T-cells, B-cells, monocytes, and neutrophils up to 2 years post sampling. Results obtained from frozen stabilized and fresh blood samples were compared for validation purposes and correlated to differential blood counts from a conventional hematology analyzer. Clinical applicability of the new technique was verified by using samples from a treatment study for feline infectious peritonitis (FIP). Using the new SMT protocol on retained samples, it could be demonstrated that long-term storage of these SMT tubes is also possible. In summary, the newly adapted SMT protocol proved suitable for performing flow cytometry analysis on stored feline whole blood samples, thus opening up new avenues for veterinary research on a variety of aspects of clinical interest.

Evaluation of Stored Whole Blood and Monitoring the Health of Dogs After Transfusion Using Fresh Whole Blood, Stored Whole Blood, and Packed Red Blood Cells

Blood products have been widely used in emergencies and treatment, necessitating optimal storage conditions to maintain quality. The current study aimed to evaluate the blood quality during storage, transfusion effectiveness, and reactions during and after transfusion in dogs. Five Greyhounds, including three males and two females aged 2.5 years old, and with 25-30 kg bodyweight, were selected and randomly labeled N1, N2, N3, N4, and N5. Fresh whole blood, stored whole blood, and packed red blood cells from the samples dogs were used for transfusion in the study. The investigated parameters were total protein (TP), aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), mean corpuscular volume (MCV), total carbon dioxide (tCO2¬), creatine kinase (CK), creatinine (CREA), blood urea nitrogen (BUN), glucose (GLU), white blood cells (WBC), red blood cells (RBC), hematocrit (HCT), plaletes (PLT), calcium (Ca), phosphorus (P), chloride (Cl), manganese (Mg), sodium (Na), and potassium (K). The results indicated that all parameters of stored blood samples were in the normal range during 28 days of storage in a refrigerator at 2-4°C. However, some parameters (TP, AST, ALT, ALP, LDH, MCV, tCO2, and K) increased, while others (CK, CREA, BUN, GLU, WBC, RBC, HCT, PLT, Ca, P, Cl, Mg, and Na) decreased during the storage period, especially Ca, P, and Na were below the normal range. All dogs indicated no reactions during and 5 hours after transfusion. However, dogs had symptoms of inappetence and mild diarrhea in 1-2 days after transfusion. Dogs received fresh whole blood recovered on day 3, while dogs of the stored blood recipient group recovered on day 4. By day 5, all dogs were healthy with no abnormal signs. The findings indicated the presence of hematological and biochemical alterations in stored blood, highlighting the importance of considering transfusion of stored blood for patients with critical medical conditions.

Acute fetal leukemia: When should it be suspected? What assessment should be performed? A case series and review of literature.

Acute fetal leukemia is rare and characterized by a very poor prognosis. The aims of this study were to identify cases of acute fetal leukemia and to describe ultrasound and fetopathological findings that should lead to a suspicion of this diagnosis, as well as the investigations required to confirm it. A national retrospective study was conducted. Clinical data, prenatal ultrasounds and postmortem findings of fetal acute leukemia cases were collected and analyzed. We collected seven cases: four in utero fetal deaths, two neonatal deaths and one termination of pregnancy. Prenatal ultrasounds showed fetal hydrops (42.9%) associated with hepatosplenomegaly (100%). In addition, post-mortem examination (n=6) suggested a Down syndrome in one case and showed other organomegaly (83.3%) due to blastic infiltration, mainly in the liver, along with extrahepatic multivisceral hematopoiesis. Immunostainings allowed to specify the type of leukemia (71.4%). In one case, diagnosis was made on blood smear and flow cytometry was performed on fresh blood samples. All cases corresponded to acute myeloid leukemia. Karyotype was abnormal in 4 cases (66.7%), including one free trisomy 21, two mosaic trisomy 21 and one chromosome 15 deletion. GATA1 gene mutations were identified in two cases: one mosaic trisomy 21 and one with normal karyotype. Any hepatosplenomegaly associated with fetal hydrops and a negative immune, infectious, and metabolic work-up, should suggest acute fetal leukemia and prompt additional investigations.

MiR-33a-5p in stored red blood cells regulates genes of innate immune response and promotes inflammation.

Blood transfusion is a common therapeutic procedure in hospitalized patients. Red blood cell (RBC) units undergo various biochemical and morphological changes during storage (storage lesion). miRNAs have been studied intensively regarding cellular metabolic processes, but the effect of miRNAs on blood storage is not well defined. We performed bioinformatics analysis on the public data set of miRNA expression of RBC based on R language, and performed the Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis on the target genes of differentially expressed miRNA. The expression of miRNA differential genes in blood samples stored at different times was verified by qRT-PCR. Next, we used ELISA and qRT-PCR to verify the expression of IL-1β, IL-6, IL-12 and TNF-α in blood at day 1 and day 42. In addition, in vitro, we transfected macrophages with overexpressed miRNA, and the effects of overexpressed miRNA on macrophage polarization and the release of inflammatory factors were verified by flow cytometry and qRT-PCR and ELISA. This study combined bioinformatics analysis and experiments to discover the differentially expressed miRNAs in long-term stored blood. The results showed that compared to fresh blood samples, the inflammatory factors were significantly doubled by ELISA, as well as the higher mRNA expression at 42 day. Experimentally verified that miR-33a-5p promoted the M1 type macrophage polarization and increased the release of related inflammatory factors through PPARα/ACC2/AMPK/CPT-1a axis regulation. This study elucidates a potential mechanism of inflammatory factor accumulation in long-term stored blood, providing a theoretical basis and a potential target to prevent transfusion-related adverse reactions.

Research on the Dilemma and Countermeasures of Migrant Workers Returning Home to Start Their Own Business under the Background of Rural Revitalization

With the comprehensive and orderly development of the rural revitalization strategy, it needs to inject fresh blood, and returning migrant workers just add new vitality to this. In order to deeply study the difficulties encountered by migrant workers returning to their hometown to start their own businesses under the background of rural revitalization and the countermeasures, this paper adopts the field interview method, literature research method and questionnaire survey method to carry out a survey in Bengbu City, Anhui province. Through the investigation, it is found that the difficulties faced by migrant workers returning home to start their own businesses are as follows: the publicity channels are not smooth; The implementation of supportive policies is lacking; Difficulties in financing loans; Rural land circulation is difficult; The cultural quality of migrant workers is not high. In view of this, the paper puts forward the countermeasures to promote the problem of migrant workers returning home to start their own businesses: build an entrepreneurial information platform; Increase policy support; Smooth loan financing channels; Reasonable transfer of land resources; Conduct entrepreneurship training and counseling. The purpose of this paper is to guide migrant workers to return home and start their own businesses reasonably and effectively, so as to promote the implementation of rural revitalization strategy.

Evaluation of Fluorescent Spot Test with Dried Blood Spots for Glucose-6-Phosphate Dehydrogenase Deficiency Screening in A Malaria-Endemic Area

The utilization of dried blood spot (DBS) samples for screening glucose-6-phosphate dehydrogenase (G6PD) deficiency remains limited in resource-constrained and malaria-endemic regions of Thailand. This study evaluated the feasibility of employing DBS for G6PD deficiency screening among 242 participants (118 males and 124 females) who provided fresh whole blood (FWB) and DBS samples for analysis. The G6PD deficiency diagnostic performances of the fluorescent spot test using DBS (FST-DBS) were compared to those using FWB (FST-FWB). The G6PD gene variant was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using DBS samples. The FST-DBS showed no significant difference in sensitivity (95.0% vs 100.0%) but superior specificity (100.0% vs 80.2%) and a positive predictive value (100.0% vs. 50.0%) compared to FST-FWB. The PCR-RFLP revealed a G6PD mutation incidence rate of 16.5% (11.1% in males and 5.4% in females). Among 40 DBS samples, 39 (97.5%) were the Mahidol (487G>A) while a sample (2.5%) which both FST-DBS and FST-FWB showed positive results had an unidentified variant. Therefore, FST-DBS is an alternative format for G6PD deficiency screening in malaria-endemic regions, particularly where resources are limited.

BIOM-01. CARBONIC ANHYDRASE IX INTERPLAY WITH LOCAL AND SYSTEMIC IMMUNE RESPONSE IN PEDIATRIC BRAIN TUMORS

Abstract BACKGROUND Brain tumors (BT) remain the leading cause of death among childhood cancers. There is an urgent need to identify novel targets in these patients. BT progression depends on the adaptive responses to hypoxia. Carbonic anhydrase IX (CAIX) helps cancer cells survive in a hypoxic environment and can be potentially as a therapeutic target. The role of CAIX in the interplay between the tumor and immune cells requires in-depth investigation to design effective therapies. METHODS Fresh BT, blood, and intraoperative cerebrospinal fluid (iCSF) samples were collected prospectively from pediatric patients, who underwent brain tumor surgery at Vilnius University Hospital Santaros Klinikos from March 2022 to December 2023. All tumor types were included. Immunohistochemistry (IHC) and flow cytometry (FC) were performed to analyze the expression of CAIX and selected immunological markers in the fresh tumor, iCSF and peripheral blood mononuclear cells (PBMCs). Soluble CAIX content in serum and iCSF was assessed by ELISA. In addition, a retrospective analysis of previously diagnosed pediatric glioma specimens was performed by IHC. RESULTS Twelve patients with heterogeneous BT types have been enrolled prospectively and 17 patients with glioma were reviewed retrospectively. Glioma accounted for 72% (21/29) of all samples. IHC showed that 57% of gliomas were positive for CAIX as well as a moderate correlation between the CAIX score and the expression of CD44, CD206, and CCR2 markers. PD-1/PD-L1 expression was negligible. In the prospective cohort, 83% (10/12) of BT tissue was positive for CAIX by IHC and FC. Soluble CAIX was detectable in 40% of iCSF samples, but not in serum. We found a correlation trend of CAIX expression in the fresh tumor cells and iCSF with increased CCR2 expression on CD45+ PBMCs. CONCLUSIONS This interim analysis showed a correlation between CAIX tumor expression and immune biomarkers, prompting further investigation by enrolling more patients.

Mitochondrial Dysfunction in Sporadic Amyotrophic Lateral Sclerosis Patients: Insights from High-Resolution Respirometry.

Amyotrophic lateral sclerosis is a severe neurodegenerative disease whose exact cause is still unclear. Currently, research attention is turning to the mitochondrion as a critical organelle of energy metabolism. Current knowledge is sufficient to confirm the involvement of the mitochondria in the pathophysiology of the disease, since the mitochondria are involved in many processes in the cell; however, the exact mechanism of involvement is still unclear. We used peripheral blood mononuclear cells isolated from whole fresh blood from patients with amyotrophic lateral sclerosis for measurement and matched an age- and sex-matched set of healthy subjects. The group of patients consisted of patients examined and diagnosed at the neurological clinic of the University Hospital Martin. The set of controls consisted of healthy individuals who were actively searched, and controls were selected on the basis of age and sex. The group consisted of 26 patients with sporadic forms of ALS (13 women, 13 men), diagnosed based on the definitive criteria of El Escorial. The average age of patients was 54 years, and the average age of healthy controls was 56 years. We used a high-resolution O2K respirometry method, Oxygraph-2k, to measure mitochondrial respiration. Basal respiration was lower in patients by 29.48%, pyruvate-stimulated respiration (respiratory chain complex I) was lower by 29.26%, and maximal respiratory capacity was lower by 28.15%. The decrease in succinate-stimulated respiration (respiratory chain complex II) was 26.91%. Our data confirm changes in mitochondrial respiration in ALS patients, manifested by the reduced function of complex I and complex II of the respiratory chain. These defects are severe enough to confirm this disease's hypothesized mitochondrial damage. Therefore, research interest in the future should be directed towards a deeper understanding of the involvement of mitochondria and respiratory complexes in the pathophysiology of the disease. This understanding could develop new biomarkers in diagnostics and subsequent therapeutic interventions.

Pulmonary hemorrhage in oncologic patients – a diagnostic algorithm

The OBJECTIVE was to develop a diagnostic algorithm in oncologic patients with pulmonary hemorrhage.METHODS AND MATERIAL. A retrospective single-center study of the medical records of 258 patients who met the inclusion criteria was conducted. Inclusion criteria: age over 18 years, oncologic disease of the chest organs complicated by pulmonary hemorrhage. Inclusion criteria: age over 18 years, oncologic disease of the chest organs complicated by pulmonary hemorrhage. Exclusion criteria: age under 18 years, pulmonary hemorrhage of non-oncologic etiology. Pulmonary hemorrhage was considered to be the discharge of any volume of fresh blood or clots from the tracheobronchial tree, which was determined on the basis of anamnesis, clinical manifestations in the hospital, endoscopic examination. Routine diagnostic methods of investigation, such as chest radiography, chest computed tomography (including with contrast enhancement), fibrotracheobronchoscopy, bronchial arteriography were evaluated.RESULTS. The sensitivity for diagnosing the oncologic process in chest radiography, computed tomography, and fibrotracheobronchoscopy, was 84.7 %, 98.4 %, and 94.4 %, respectively. The sensitivity of fibrotracheobronchoscopy to detect the bleeding itself in oncologic patients reached 31.3 %, and the sensitivity of bronchial arteriography to diagnose the source of bleeding was 87.7 %.CONCLUSION. In pulmonary hemorrhage of oncologic genesis, the combination of fibrotracheobronchoscopy with computed tomography has sensitivity in 100 % of cases in determining the source and localization of the pathological process. Bronchial arteriography allows to perform endovascular hemostasis. It is necessary to conduct multicenter studies in order to develop and implement a unified algorithm assessing all etiopathogenetic features of pulmonary hemorrhage in oncologic patients.

DEVELOPING A THROMBOELASTOGRAPHY ASSAY IN ELASMOBRANCHS.

Thromboelastography (TEG) is a hemostatic assay evaluating clot initiation time, kinetics, strength, and extent of fibrinolysis. Hemostatic assays in nonmammalian species have been less extensively studied because of lack of taxon-specific reagents and unique physiology. Hemostatic or hemorrhagic disease has been described postmortem in elasmobranchs, but antemortem detection of coagulopathies is limited in this taxon. The study aimed to establish an elasmobranch TEG protocol to improve hemostatic evaluation and facilitate advanced treatment options for animals under human care. Multiple clotting initiators were assessed for efficacy with frozen-thawed citrated plasma, fresh citrated plasma, and fresh whole citrated blood: RapidTEGTM, citrated kaolin, Reptilase®, and species brain-derived thromboplastin prepared by two different methods. Initial evaluation found plasma samples clot inconsistently, but TEG analyses using fresh whole blood consistently led to measurable TEG reactions using multiple clotting initiators. The most reliable elasmobranch TEG results were observed using citrated fresh whole blood and the RapidTEG clot initiation reagent.

Designing an experimental method for assessing biocompatibility of circuit coatings using biomarkers for platelet activation during cardiopulmonary bypass.

Cardiopulmonary bypass is an essential component of cardiothoracic surgeries. However, significant complications such as systemic inflammatory response syndrome (SIRS) resulting from cardiopulmonary bypass (CPB) are a common occurrence due to contact between circulating blood and foreign surfaces that leads to platelet activation. It is suggested that different available CPB circuit coatings can potentially reduce platelet activation. However, there have been no published evidence-based reports confirming these claims. In addition, there is no well-established protocol for studying platelet activation biomarkers during CPB in vitro in a laboratory setting. CPB was simulated in the laboratory using bovine blood in two different types of coated CPB circuits: Trillium® Biosurface by Medtronic, and XcoatingTM Surface by Terumo. Fresh bovine blood samples were collected and circulated through the CPB circuit following the standard protocol used in the operation rooms. Blood samples were then collected at 5min, 30min, and 55min during the circulation. Blood plasmas were separated and subjected to enzyme-linked immunosorbent assay to measure most established platelet activation markers P-selectin, Platelet Factor 4 (PF4), Glycoprotein IIb/IIIa (GPIIb/IIIa), and β-thromboglobulin (β-TG) at different time points. The biomarker values at 30min and 55min were compared to the base values at 5min for each type of CPB circuit. The results of the means from all measured biomarkers showed data measurements that indicated no significant variability within each coating. All collected data points fell within±2 SD of the means, which was considered acceptable variations across technical replicates. Conclusion: In this study, we were able to establish an in vitro protocol in the laboratory setting that is precise and reliable with minimum intra-variability. This established protocol will allow for future studies in which different coated CPB circuits can be compared for their effectiveness in blocking platelet activation during the CPB.

Tislelizumab plus concurrent chemoradiotherapy versus concurrent chemoradiotherapy for elderly patients with inoperable locally advanced esophageal squamous cell carcinoma: A multicenter, randomized, parallel-controlled, phase II clinical trial.

TPS4183 Background: There is few high-level prospective randomized studies and international consensus guideline on treatment in elderly patients with inoperable locally advanced esophageal squamous cell carcinoma (LAESCC). Double-agent intravenous chemotherapy concurrent with radiotherapy is the standard of care for LAESCC. However, it’s based on clinical trials that only enrolled patients ≤ 75 years old and infeasible in elderly patients. Oral S-1 plus concurrent radiotherapy for elderly ESCC showed tolerable toxicity and survival benefit compared with radiotherapy alone. While, the 3-year overall survival (OS) rate is limited about 40%. Therefore, it urgently needs find new treatment strategies to further improve the efficacy in elderly ESCC patients. Tislelizumab, the antii-PD-1 antibody, has recently been proved in the first- and second-line standard treatment for advanced ESCC. It shows mild toxicities and might be well-tolerated in the elderly. Here, we conducted the first clinical trial to assess the efficacy and safety of tislelizumab plus concurrent chemotherapy (CCRT) versus CCRT for elderly patients with inoperable LAESCC. Methods: The study is an open label, multicenter, parallel-controlled, investigator-initiated phase II clinical trial in elderly patients with inoperable LAESCC. We plan to enroll 136 elderly patients with LAESCC who will be randomly divided into two groups: tislelizumab combined with CCRT group (tislelizumab + radiotherapy + S-1) and CCRT group (radiotherapy + S-1). Tislelizumab is given intravenously on the first day of CCRT at the dose of 200 mg once every 3 weeks (Q3W). Radiotherapy is delivered at 50.4 Gy/1.8 Gy/28 fraction (5 times per week). S-1 is orally given at the dose of 40-60 mg from day1 to day14 and day 22 to 35. We will evaluate the efficacy and safety during the treatment and follow-up period until disease progression, death or the end of the trial. Considering the median progression free survival (PFS) is 16 months in elder receiving CCRT in history studies, we expect the median PFS would be 28 months at the hazard ratio of 0.57. The final sample size consists in 136 patients by 1:1 randomized in the two groups with 80% power and one-sided alpha and calculating for a 10% dropout rate. The primary endpoint is PFS. Secondary endpoints include OS, objective response rate, duration of objective response and severity of adverse events. Fresh or archival tumor tissues and peripheral blood samples will be used in the exploratory studies. This trial started on October 10, 2023 and has enrolled 19 patients till February 1, 2024. Clinical trial information: NCT06061146 .

A comparative study of blood cell count in four automated hematology analyzers: An evaluation of the impact of preanalytical factors.

Differential white blood cell counts are frequently used in diagnosis, patient stratification, and treatment selection to optimize therapy responses. Referral laboratories are often used but challenged with use of different hematology platforms, variable blood shipping times and storage conditions, and the different sensitivities of specific cell types. To extend the scientific literature and knowledge on the temporal commutability of blood samples between hematology analyzers, we performed a comparative ex-vivo study using four of the most utilized commercial platforms, focusing on the assessment of eosinophils given its importance in asthma management. Whole blood from healthy volunteers with and without atopy (n = 6+6) and participants with eosinophilic asthma (n = 6) were stored under different conditions (at 4, 20, 30, and 37°C, with or without agitation) and analyzed at different time points (3, 6, 24, 48 and 72h post-sampling) in parallel on the Abbott CELL-DYN Sapphire, Beckman Coulter DxH900, Siemens ADVIA 2120i and Sysmex XN-1000V. In the same blood samples, eosinophil-derived neurotoxin (EDN), eosinophil activation and death markers were analyzed. All platforms gave comparable measurements of cell differentials on fresh blood within the same day of sampling. However, by 24 hours, significant temporal and temperature-dependent differences were observed, most markedly for eosinophils. None of the platforms performed perfectly across all temperatures tested during the 72 hours, showing that handling conditions should be optimized depending on the cell type of interest and the hematology analyzer. Neither disease status (healthy vs. asthma) nor agitation of the sample affected the cell quantification result or EDN release. The eosinophil activation markers measured by flow cytometry increased with time, were influenced by temperature, and were higher in those with asthma versus healthy participants. In conclusion, hematology analyzer, time window from sampling until analysis, and temperature conditions must be considered when analyzing blood cell differentials, particularly for eosinophils, via central labs to obtain counts comparable to the values obtained in freshly sampled blood.