Search Results (182)

Immunotherapy has revolutionized cancer treatment; however, the availability of cost-effective blood-based biomarkers for prognostic and predictive factors of immune treatment in patients with solid tumors remains limited. Due to low cost and easy accessibility, blood-based biomarkers should constitute an essential component of studies to optimize and monitor immunotherapy. Currently available markers that can be measured in peripheral blood include total monocyte count, myeloid-derived suppressor cells (MDSCs), regulatory T cells (Tregs), relative eosinophil count, cytokine levels (such as IL-6, IL-8, and IL-10), lactate dehydrogenase (LDH), C-reactive protein (CRP), soluble forms of CTLA-4 and PD-1 or PD-L1, as well as circulating tumor DNA (ctDNA). In our mini-review, we discuss the latest evidence indicating that routinely accessible peripheral blood parameters—such as the neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), platelet-to-lymphocyte ratio (PLR), and rheological parameters, which so far have been rarely considered for such an application, may be used as non-invasive biomarkers in cancer immunotherapy. Rheological parameters such as whole blood viscosity are influenced by several factors, such as hematocrit, aggregability and deformability of erythrocytes, and plasma viscosity, which is largely dependent on plasma proteins. Especially in cases where the set of symptoms indicates a high probability of hyperviscosity syndrome, blood rheological tests can lead to early diagnosis and treatment. Both biochemical and rheological parameters are prone to become novel and future standards for assessing immunotherapy among patients with solid tumors. Full article

Ocular flutter is an uncommon ophthalmic finding that may indicate paraneoplastic phenomena, and it is clinically characterized by intermittent bursts of conjugate, horizontal saccades without an intersaccadic interval. Ocular flutter must be differentiated from opsoclonus, which, although also characteristic of certain paraneoplastic syndromes, is instead defined by multidirectional saccades on both the horizontal and vertical planes. This report describes a very rare presentation of anti-Ri syndrome in a patient with an undiagnosed breast cancer, presenting with ocular flutter, dizziness, blurred vision, photophobia, and vomiting. Comprehensive evaluations, including contrast-enhanced brain Magnetic Resonance Imaging (MRI), brain Computed Tomography (CT) scan, ophthalmological assessment, viral serology, complete blood count and thyroid, renal coagulation, hepatic function assessments, vitamin D and B12 levels, were all normal. Upon excluding other potential etiologies for the neurological symptoms, a paraneoplastic origin was considered. Serological tests confirmed the presence of anti-Ri onconeural antibodies, and a whole-body CT scan identified nodules in the right breast. Despite surgical excision of the primary tumor and subsequent medical therapy, there was no improvement in the neurological symptoms. Follow-up evaluations at 2 months, 6 months, 1 year and 2 years revealed persistent vestibular and neurological symptoms, with serum tests remaining positive for anti-Ri antibodies and no clinical or radiological evidence of neoplastic recurrence. Full article

Hemoglobin disorders are among the most common inherited diseases worldwide. Their clinical manifestations range from anemia to more severe forms associated with neurological impairments. These complications can result as secondary consequences of the disease’s clinical manifestations or be directly linked to genetic mutations. In this study, we present two families with neurological impairments who were referred to us for complementary hematological and biochemical analyses. Complete blood count, methemoglobin level, and methemoglobin reductase activity were assessed. Molecular analyses were performed using whole-exome sequencing, and the segregation of the identified mutations was confirmed with direct sequencing. Their pathogenicity and conservation were evaluated using various bioinformatics tools. Clinical and hematological findings suggested X-linked alpha-thalassemia/impaired intellectual development syndrome in the first family and recessive congenital methemoglobinemia type II in the second. This was confirmed by the identification of pathogenic mutations ATRX: p.Arg2131Gln and CYB5R3: p.Ala179Thr, respectively. Although these variants have been previously reported worldwide, they were identified for the first time in our population. Our results contribute to the understanding of the pathogenesis of these rare disorders and provide a basis for diagnosis, treatment, and genetic counseling. The mechanisms by which these mutations contribute to neurological symptoms are discussed. Full article

Feed is the single greatest cost for cattle producers. Improvements to feed efficiency, or how animals convert feed to body weight gain, will ultimately improve producer profits. The objective of this study was to determine whether the expression of genes in the transcriptome of whole blood from heifers (n = 61) on a forage ration was related to the quantitative phenotypes for average daily gain (ADG), average daily feed intake (ADFI), and gain-to-feed (G:F). Total RNA was isolated from whole blood collected mid-study on day 42 on feed and was used for hematologic analysis and RNA-sequencing. Lymphocyte (LYM) count was negatively associated with ADG, and mean corpuscular hemoglobin concentration (MCHC) was positively associated (p < 0.05). Red blood cell (RBC) count was negatively associated with ADFI. While MCHC was positively associated with G:F, white blood cell (WBC), LYM, and basophil (BAS) counts were negatively associated with G:F (p ≤ 0.05). The model used to identify differentially expressed genes (DEG) for ADFI, ADG, and G:F included sire, breed, pen, age, and proportions of blood cell types. No genes were differentially expressed for ADFI. Three genes were identified as differentially expressed for ADG, and 17 were identified for G:F. Three of the differentially expressed genes for G:F and ADG (PLOD1, FAH, and COL1A2) had been previously associated with feed efficiency in livestock and may be useful for further validation in other populations of cattle. The negative associations between WBC and LYM and ADG and G:F may be expected since the production of WBC is an energetic process that may reduce body weight gain and efficiency. The associations between RBC hematological parameters suggest that there may be benefit for animals with higher levels of hemoglobin per RBC by improving oxygen-carrying capacity. Full article

Background/Objectives: In recent years, there has been an increase in the prevalence of eosinophilic esophagitis (EoE), in which dysphagia is one of the main symptoms. To date, there are few data on the prevalence of EoE in pediatric patients with dysphagia. The aim of this study was to determine the causes of dysphagia in children in our region. The second aim of this study was to estimate the prevalence of EoE in children with swallowing difficulties and to see if there were differences in the characteristics of dysphagia reported by children with EoE compared to children with non-EoE-related dysphagia. Methods: The 6-year retrospective analysis included patients with dysphagia who were referred to our department. Children with dysphagia were further divided into two groups: group I consisted of children with dysphagia due to EoE, while group II consisted of children with dysphagia due to other causes (non-EoE). Results: One hundred and forty-six children between the ages 0 and 17 were enrolled into the study, including thirty-seven in group I and one hundred and nine in group II. The most common causes of dysphagia were gastrointestinal diseases, followed by neurological/psychiatric disorders. The prevalence of EoE was 25.34% in the whole study group and 41.11% considering only gastrointestinal causes of dysphagia. Children in group I were more likely to have coexisting asthma, allergic rhinitis and food allergy. There was statistically significance in higher blood eosinophil count in EoE individuals. In a multivariate binominal logistic regression model, only eosinophilia and coexisting food allergy were associated with an increased risk of EoE in patients with dysphagia. Conclusions: In our study, the most common cause of dysphagia was gastroenterological diseases, especially EoE. Patients with dysphagia, comorbid allergy and peripheral blood eosinophilia should be suspected for having EoE and referred for endoscopy. Full article

New therapies are highly needed to stabilize remission in patients with acute myeloid leukemia (AML). This study investigates the value of dendritic cells derived from leukemic blasts (DC_leu) to enhance anti-leukemic immunity after T-cell-enriched mixed lymphocyte cultures (MLCs). We correlated induced anti-leukemic activity with patient data, including biological, clinical and prognostic factors. Additionally, we correlated the frequencies of DC/DC_leu and leukemic-specific T cells with the achieved anti-leukemic activity after MLC. We show that mature DC/DC_leu can be generated using the immunomodulating Kit-M, which contains granulocyte–macrophage colony-stimulating-factor (GM-CSF) and prostaglandin E₁ (PGE₁), without inducing blast proliferation from leukemic whole blood (WB) samples. Activated leukemia-specific immune and memory cells increased after MLC with Kit-M-pretreated WB, leading to improved blast lysis. Enhanced anti-leukemic activity positively correlated with the frequencies of generated DC/DC_leu, proliferating leukemic-specific T cells and memory T cells, but not with leukemic blast counts, hemoglobin levels or platelet counts at diagnosis. No correlation was found between improved blast lysis and patients’ prognostic data, including age, gender, ELN risk groups, disease stage and response to induction chemotherapy. These findings underscore the potential of DC/DC_leu to evoke robust immune responses and potential immunological memory against AML. Overall, this innovative approach could pave the way for the development of improved immunotherapeutic strategies that function in vivo. Full article

Whole blood viscosity is a test for blood stasis and is an ideal evidence-based pathology parameter that is largely undervalued and retrogressing in clinical utilization. Coagulation profiles as indices of haemostasis are available but limited to central or referral laboratories and often involve long turn-around time. It is therefore important to study the correlation between the index of stasis and indices of haemostasis. Objective: To investigate the correlation of index of stasis with indices of haemostasis. Method: The clinical laboratory observational research method, using archived pathology data. Indices of haemostasis including activated partial thromboplastin time (APTT) and prothrombin time (PT), the international normalization ratio (INR), and plasma D-dimer were evaluated. On the other hand, the index of blood stasis used was the estimated whole blood viscosity (eWBV) and derived haematocrit and serum protein levels. All (N = 193) tests were collected within a calendar year from the same pathology service, and further, for the correlation, each set of variables from the same blood sample collection was used. Results: The haemostasis data are skewed (skewness > 2.0), while eWBV and platelet count are normal (skewness < 2.0). Haemostasis indices have an inverse association with eWBV (p < 0.001). The concordance and correlation of eWBV with platelet count is positive, weak, and significant (p < 0.001), but negative and negligible with PT and APTT. Conclusion: There are limitations to the possible correlation between eWBV and haemostasis indices. However, haemostasis indices have inverse associations with eWBV, and the latter can aid in the evaluation of haemostasis hence could be utilized as an alternative or complementary test to haemostasis tests. Research may normalize skewed data to obtain better correlation; therefore, further study is required to advance discourse, giving cognizance to clinical practice. Full article

Sixty Holstein cows were enrolled at −76 days from calving (DFC) and classified based on the daily SCC during the previous week from an automated milking system. The separation thresholds for low (L, n = 46) and high (H, n = 14) classifications were 100 K/mL for primiparous and 200 K/mL for multiparous cows. Cows were then assigned to two homogeneous groups to receive diets supplemented with 19 g/d of a Saccharomyces cerevisiae fermentation product (TRT; NutriTek, Diamond V, Cedar Rapids, IA, USA) or without supplementation (CTR) until 60 DFC. Cows were dried off at −56 DFC and monitored for disease incidence, milk yield and composition, plasma metabolic profile, and whole blood count from −76 to 60 DFC. Data were analyzed utilizing ANOVA and mixed models for repeated measures. During the dry period, TRT cows had greater plasma thiol and albumin compared to CTR. TRT-L cows had greater plasma protein and globulin than CTR-L. TRT-H cows had heightened hematocrit; reduced plasma globulin and haptoglobin; and higher albumin, albumin to globulin ratio, and thiol than CTR-H. TRT-H cows had greater concentrations of leukocytes and lymphocytes and lower plasma protein and ceruloplasmin at −54 DFC; lower reactive oxygen species to ferric ion-reducing antioxidant power ratios at −44 DFC; and greater concentrations of lymphocytes and plasma gamma glutamyl transferase at −7 DFC than CTR-H. After calving, TRT cows had a lower incidence of mastitis and higher butterfat, as well as greater plasma haptoglobin and aspartate amino transferase (AST) and reduced Mg compared to CTR. TRT cows had lower SCC between 1 and 7 DFC and a greater ECM between 41 and 60 DFC compared to CTR. TRT-H cows had lower SCC between 1 and 7 DFC and greater hemoglobin and plasma AST than CTR-H. Ameliorated immune system functions due to Saccharomyces cerevisiae fermentation product administration lowered the SCC in TRT-H cows and prevented the onset of new intramammary infections across both L and H SCC groups, supporting the improved productive performance of dairy cows. Full article

Objectives: There are still limited or lacking data on the association of trace elements (TEs) with polycystic ovary syndrome (PCOS). This case–control study aimed to determine levels of essential TEs (manganese (Mn), copper (Cu), zinc (Zn), selenium (Se), molybdenum (Mo)) and non-essential TEs (arsenic (As), cadmium (Cd), mercury (Hg), lead (Pb)) in urine, whole blood, and serum to investigate a possible association with kidney and liver function, endocrine and metabolic parameters, and environmental and lifestyle sources of potential exposure and provide possible recommendations. Methods: In our case–control study, women with PCOS (n = 35) and healthy controls (n = 35) underwent clinical and ultrasonographic examination, filled in questionnaires targeting general, lifestyle, and environmental information, and provided fasting venous blood samples and first morning urine for biochemical, hormonal, and TE analysis. Multiple linear regression models were used to evaluate the association between TE levels and data obtained through questionnaires. Results: In women with PCOS, lower Mo levels in whole blood (p = 0.024) and serum (p = 0.011) and higher serum Cu levels (p = 0.026) were detected when compared to healthy controls. Results of this study show that amendments in Cu and Mo levels might be related to altered kidney and liver function and disrupted hormonal balance in PCOS women. Cu levels positively correlated with leukocyte count. There was a negative correlation of Mo levels with proteinuria and luteinizing hormone levels. Regarding liver function, Mo negatively correlated with urinary bilirubin levels, and there was a positive association with alanine and aspartate aminotransferase, respectively. Dietary supplement consumption and certain diet habits appeared to be important predictors of exposure to Cu (beef consumption) or Mo (cereal and boiled vegetable consumption) and modify Mo and Cu levels in women. Conclusions: Concentrations of the chemical elements Mo and Cu in biological samples of women appear to be related to PCOS and nutrition. To our knowledge, this is a novel finding for Mo. Additional research is needed to provide more insights into the causality of the PCOS relationship with Mo and Cu in humans. Full article

Echis ocellatus envenomings are a public health problem in West Africa, leading to bleeding and hypocoagulability. The aim of this study was to assess the hemostasis disorders associated with E. ocellatus envenoming. Envenomed patients with an abnormal whole blood clotting test (WBCT) were prospectively included at Tanguiéta, Benin. A WBCT with a sequential reading (i.e., at 20, 30, and 60 min), viscoelastic analysis (VA) using the Quantra analyzer, and blood count were performed on admission. VA and the WBCT were also assessed at 4, 8, 12, 24, 48, and 72 h after antivenom administration. Nineteen patients were included. On admission, the main results were an absence of a clot on VA and a slight decrease in platelets. Clot time gradually decreased over time while clot stiffness, fibrinogen, and platelet contributions to stiffness increased. Sequential reading improved the sensitivity of the WBCT. At H48, all patients with recurrence bleeding after antivenom administration had an abnormal WBCT while patients with a normal WBCT never had bleeding during their follow-up. VA allows the identification of various hemostasis disorders. Hypofibrinogenemia was the main disorder that persisted for several days after treatment. A WBCT with a sequential reading is an effective alternative for monitoring hypocoagulability in the absence of a laboratory. Full article

Myeloid chimerism better reflects donor stem cell engraftment than whole-blood chimerism in assessing graft function following allogeneic hematopoietic stem cell transplant (HCT). We describe our experience with 130 patients aged younger than 18 years, treated with allogeneic HCT using bone marrow or PBSC from HLA-matched donors for non-malignant diseases, whose pre-transplant conditioning therapy included alemtuzumab and who were monitored with lineage-specific chimerism after transplant. At 6 years post-transplant, overall survival (OS) was 91.1% and event-free survival (EFS) was 81.5%, with no grade III-IV acute GvHD or chronic GVHD observed. Recipient T-cells did not contribute to graft loss. Mixed T-cell chimerism (MC) did not affect EFS, and there was no connection between T-cell chimerism and myeloid chimerism in patients with MC or graft loss. MC significantly correlated with virus infection; more children with MC were CMV seropositive than those with complete chimerism (CC). Additionally, MC was more common in patients with CMV viramia post-transplant. CD8 T-cell reconstitution was affected by viral reactivation, including CMV, with CD8 T-cell counts higher in the MC group than in the CC group. Mixed T-cell chimerism is due to autologous, virus-specific, predominantly CD8, T-cell expansion, and is protective and not deleterious to the recipient. Full article

The cost-effectiveness of whole exome sequencing (WES) remains controversial due to variant call variability, necessitating sensitivity and specificity evaluation. WES was performed by three companies (AA, BB, and CC) using reference standards composed of DNA from hydatidiform mole and individual blood at various ratios. Sensitivity was assessed by the detection rate of null–homozygote (N–H) alleles at expected variant allelic fractions, while false positive (FP) errors were counted for unexpected alleles. Sensitivity was approximately 20% for in-house results from BB and CC and around 5% for AA. Dynamic Read Analysis for GENomics (DRAGEN) analyses identified 1.34 to 1.71 times more variants, detecting over 96% of in-house variants, with sensitivity for common variants increasing to 5%. In-house FP errors varied significantly among companies (up to 13.97 times), while DRAGEN minimized this variation. Despite DRAGEN showing higher FP errors for BB and CC, the increased sensitivity highlights the importance of effective bioinformatic conditions. We also assessed the potential effects of target enrichment and proposed optimal cutoff values for the read depth and variant allele fraction in WES. Optimizing bioinformatic analysis based on sensitivity and specificity from reference standards can enhance variant detection and improve the clinical utility of WES. Full article

Differences in cell count and growth factor expression between first- and second-generation autologous platelet concentrates (APCs) have been well described. The debate over which formula best supports wound healing in various surgical procedures is still ongoing. This study aims to assess the whole proteome assembly, cell content, immunological potential and pro-angiogenic potential of second-generation APC, Platelet-Rich Fibrin (PRF) vs. first-generation APC, Platelet-Rich Plasma (PRP). The global proteome of the APCs was analyzed using nano-liquid chromatography mass spectrometry. Blood cell concentrations were determined by an automated cell counter. The effect of APCs on macrophage polarization was analyzed by flow cytometry. A yolk sac membrane (YSM) assay was used to monitor the neo-vessel formation and capillary branching in vivo. Cell count analysis revealed a higher number/concentration of leukocytes in PRF vs. PRP. Incubation of macrophages with PRP or platelet-free plasma (PFP) did not induce a significant pro-inflammatory state but led to a shift to the M0/M2 phenotype as seen in wound healing for all tested formulas. Label-free proteomics analysis identified a total of 387 proteins from three biological replicates of the respective designated groups. PRF induced increased formation of neo-vessels and branching points in vivo in comparison to PRP and PFP (each p < 0.001), indicating the enhanced pro-angiogenic potential of PRF. Overall, PRF seems superior to PRP, an important representative of first-generation formulas. Inclusion of leucocytes in PRF compared to PRP suggested rather an anti-inflammatory effect on macrophages. These results are important to support the versatile clinical applications in regenerative medicine for second-generation autologous platelet concentrates to optimize wound healing. Full article

The federally threatened eastern indigo snake (EIS; Drymarchon couperi) is an active ophiophagous snake once found throughout the southeastern US that is now restricted to southeastern Georgia and peninsular Florida. There are concerns regarding the potential impact of overconditioning or nutrient imbalances on the reproductive fitness of breeding programs due to the occurrence of dystocia in nulliparous dams and the differing nutritional profiles of domestic and free-range prey species. We examined the blood cell counts, plasma biochemistry, and circulating plasma levels of nutrients in snakes consuming standard or experimental diets over a one-year period. Treatments included mixed whole laboratory animal prey (rodents, birds), whole prey ground into sausage, or a sausage with similar nutrient profiles measured in prey found in free-ranging EIS stomach contents. Plasma concentrations of vitamin E (maximum = 0.80 mg/mL) and selenium (maximum = 371 ng/mL) were within range of and exceeded values reported in free-ranging EIS (0.0365 mg/mL and 107.45 ng/mL), while plasma vitamin D₃ concentrations (maximum = 64.1 ng/mL) were typically below minimum values observed in free-ranging EIS (46 ng/mL). Additional dietary studies initiated on juvenile subjects throughout reproductive maturity would provide an ideal experimental design for studying the linkage between reproductive health and nutrition. Full article

Background: In response to inflammation, the absorption of nutritional iron is restricted. Since the pathophysiological significance of the presence and uptake of iron in chronic inflammation is still unknown, we tested the effect of a low iron diet on the clinical course of arthritis in the mouse model of collagen-induced arthritis (CIA). Methods: Six- to eight-week-old male DBA/1 mice were fed either a normal (51 mg/kg) or a low iron diet (5 mg/kg) starting four weeks before the first immunization. From day 4 after the second collagen booster made on day 25, the development of arthritis was regularly monitored until the end of the experiment (day 34), using a standard clinical arthritis score. Concentrations of mouse anti-bovine and anti-mouse collagen type 2 IgG antibodies were measured by ELISA; blood cell counts were performed and mediators of inflammation, tissue matrix degradation, oxygenation and oxidative stress were measured in the mouse sera of both diet groups at the end of the experiment by bead-based multiplex assay. Fe²⁺, Fe³⁺, oxidized and reduced glutathione (GSH and GSSG) and malondialdehyde (MDA) were quantified in whole paw tissue by ELISA. Quantitative PCR was performed in the tissues for glutathione peroxidase 4 and other key regulator genes of iron metabolism and ferroptosis. We used nonparametric tests to compare cross-sectional data. Nonlinear regression models were used for longitudinal data of the arthritis scores. Results: Mice fed a low iron diet showed a significantly less severe course of arthritis compared to mice fed a normal iron diet (p < 0.001). The immune response against bovine and mouse type 2 collagen did not differ between the two diet groups. Mice fed a low iron diet exhibited significantly lower serum levels of tissue inhibitor of metalloproteinase-1 (TIMP-1), a central regulator of inflammation and tissue matrix degradation (p < 0.05). In addition, a low iron diet led to a significant reduction in red blood cell indices, indicating restricted iron uptake and latent iron deficiency, but had no effect on hemoglobin concentrations or red blood cell counts. There were no differences between the dietary groups in Fe²⁺ or Fe³⁺ content in the paws. Based on calculation of the GSH/GSSG ratio and high MDA levels, high oxidative stress and lipid peroxidation were likewise detected in the paws of both diet groups of mice. Consequently, no differences associated with gene expression of key regulators of iron metabolism and ferroptosis could be detected between the paws of both diet groups. Conclusions: Restricted dietary iron intake alleviates immune-mediated inflammation in CIA without causing anemia. This finding suggests a promising option for dietary treatment of arthritis in inflammation. The underlying mechanism causing reduced arthritis may be linked to the complex regulatory network of TIMP-1 and appears to be independent from the local iron levels, oxidative stress and ferroptosis in the synovial tissues. Full article