Cell Count Values Research Articles

Clonal Hematopoiesis and Inflammation in the Vasculature (CHIVE), a Prospective, Longitudinal Clonal Hematopoiesis Cohort and Biorepository

Introduction Clonal hematopoiesis (CH), the expansion of a clonal population of hematopoietic cells, is an age-associated phenomenon leading to increased risk of both hematologic malignancy and non-malignant organ dysfunction. There have been significant advances in our understanding of the underlying pathophysiology of CH in recent years. However, additional studies are needed to better understand clinical trajectories and optimal care of patients with CH. To address this gap, the prospective, longitudinal CHIVE (Clonal Hematopoiesis and Inflammation in the VasculaturE) registry and biorepository was created to support multidisciplinary CH clinics. Methods Adults without active hematological malignancy who were clinically identified to have CH or who were at risk for CH were sequentially enrolled utilizing a multidisciplinary referral approach between October 2020 to April 2023 at Vanderbilt University Medical Center (VUMC). Research samples were procured at the time of routine clinical blood draws or bone marrow biopsies and stored in a newly established biorepository, equipped to store DNA, plasma, and viably cryopreserved cells to enable a spectrum of downstream applications. Specimens were sequenced on a highly cost efficient (~$6/sample) targeted next generation sequencing capture panel that identified mutations residing in the 24 most frequently mutated CHIP genes.The VUMC CH clinic, which serves as a referral pool, were included in Arm A of CHIVE (n=57). We also sought out a population at risk for CH which included patients over 18 years of age with a history of solid tumor, cardiovascular disease, renal disease, rheumatologic disease, or diabetes, who were included in Arm B (n=122). Patients from Arm A or B were included in the CH-positive (CH+) group if they met criteria for CH on the research assay at any time during their enrollment in the study. All other genotyped patients who were not found to have CH by the research assay, regardless of their clinical CH status at study enrollment, were included in the CH-negative (CH-) group.Anthropometric data and clinical assessments, including laboratories and cardiovascular studies obtained in the course of routine clinical care were extracted from the electronic health record, and recorded and maintained in the CHIVE database. Results 261 patients were approached for consent to enroll in CHIVE and 179 were ultimately enrolled. More than one half (54%) of all participants had a CH mutation (CH+) and of those, 49.5% were female. Median age of CH+ patients was 71.9 (IQR 64.0-77.5). DNMT3A (n=44) and TET2 (n=39) were the most commonly mutated genes, while variant allele fractions (VAF) ranged from 2.1 - 79.8% (Figure 1). 246 samples were serial samples collected from 89 patients at regular intervals. Significantly more CH+ patients had a diagnosis of coronary artery disease compared to CH- patients (55.7% vs 32.9%, p=0.004). CH+ patients also had increased rates of hypertension and heart failure compared to the CH- group (p<0.001, 0.035, respectively). Laboratory evaluation showed CH+ patients to have a lower median white blood cell count, hemoglobin, and platelets, and elevated blood urea nitrogen values compared to CH- patients, differences that did not meet statistical significance. 8.2% of the CH+ patients progressed to frank hematologic malignancy over the course of the 2.5-year study (Table 1). Eight of the 97 CH+ and 2 of the 82 CH- patients died over the course of the study period resulting in a trend toward decreased survival in CH+ patients (p=0.112). Conclusions We demonstrate the feasibility of a prospective, observational study of CH patients utilizing a robust referral network to support both clinical care and translational research. A well-genotyped and phenotyped cohort of CH patients will be critical to future translational research efforts and clinical trials - a key function of our study design. Early clinical findings from our cohort recapitulate large scale retrospective datasets where CH patients are at an increased risk of development of hematologic malignancy, end organ damage, and all-cause mortality. Scaling this resource in collaboration with other centers is underway and will ultimately enable the development of clinical guidelines and treatment strategies for this increasingly recognized patient population.

Comparison of the CXCR5-Antibody Drug Conjugate (ADC; VIP924) to a CD19-ADC and a CD79b-ADC in a Humanized Rec-1 Mantle Cell Lymphoma (MCL) Mouse Model

Introduction The chemokine receptor CXCR5 is highly expressed in MCL, which is a rare subtype of non-Hodgkin lymphoma. VIP924 is a first-in-class CXCR5-targeting ADC composed of a legumain-cleavable linker and a kinesin spindle protein inhibitor (KSPi) payload. We compared VIP924 with a CD19- and a CD79b- targeting ADC in a humanized mouse model of MCL. Methods Expression of CXCR5 and the B-cell targets CD19 and CD79b were analyzed on tumor samples from patients with MCL using immunohistochemistry. Slides were examined by a pathologist and scored for expression. NSG-SGM3 mice were transplanted with human hematopoietic stem cells from 4 different donors and after engraftment transplanted subcutaneously with REC-1 MCL cells into their flanks. When subcutaneous tumors reached a size of 100 mm 3, mice were randomized into 5 groups and treated with 10 mg/kg of an isotype control ADC, 3 mg/kg polatuzumab vedotin, 0.66 mg/kg loncastuximab teserine, 3 mg/kg VIP924, or 10 mg/kg VIP924 every 5 days for 4 doses. Immunophenotyping of peripheral blood was performed by flow cytometry before starting treatment (Day 0) and on Day 5 and Day 18 during the treatment. At the end of the experiment, immunophenotyping was also done on tumors, spleen, and bone marrow. Complete blood count was analyzed based on single cell evaluation (Drew Scientific HemaVet 950FS Auto Blood Analyzer). Results By immunohistochemistry CXCR5 and CD19 showed comparable high expression on patient samples, whereas CD79b expression was moderate. Treatment of humanized mice bearing REC-1 tumors with VIP924 at 10 mg/kg reduced tumor growth effectively (p<0.0001 vs. isotype control on Day 10), while treatment with polatuzumab vedotin and loncastuximab teserine showed no effects on tumor growth. This outcome was also reflected in the median survival rates for the different groups with 17 days for isotype control and polatuzumab vedotin, 14 days for loncastuximab teserine and not reached for the 10-mg/kg VIP924 group. However, polatuzumab vedotin treatment significantly reduced hCD45+ cells in peripheral blood and spleen, while loncastuximab teserine treatment showed a clear trend for reduction. VIP924 treatment in both arms had no effect on peripheral or splenic hCD45+ cells. All tested ADCs reduced the CD19+ and more mature CD19+/CXCR5+ B-cells in peripheral blood and spleen, with polatuzumab vedotin and loncastuximab teserine showing significant (p<0.0001 vs. isotype control on Day 18) effects on CD19+ cell reductions, while VIP924 treatment showed a milder reduction in CD19+ cells (p= 0.0051 for 3 mg/kg and p= 0.0407 for 10 mg/kg VIP924). Loncastuximab teserine treatment increased the amount of myeloid derived suppressor cells in the peripheral blood significantly (p= 0.0467). In contrast, VIP924 led to a significant (p< 0.0001) increase in peripheral regulatory T-cells compared with isotype control. Complete blood counts from treated mice showed a reduction of white blood cells (WBC), absolute lymphocyte count, and absolute monocyte count in loncastuximab teserine-treated animals. Loncastuximab teserine treatment also reduced red blood cell count and hemoglobin values. In VIP924-treated animals, no effects on WBC, red blood cell count, or hemoglobin values were observed. All treatments showed no detrimental effects on body weights of the mice. Conclusion CXCR5 is a highly attractive target in B-cell malignancies such as diffuse large B-cell lymphoma and MCL due to high CXCR5 surface expression. VIP924, a CXCR5-targeting ADC, showed high potency and superiority to other B-cell-targeted ADCs in a humanized mouse model of MCL. In this MCL model in humanized mice, VIP924 at 10 mg/kg treatment showed a significant reduction in tumor growth and improvement in survival, while polatuzumab vedotin and loncastuximab teserine had no effect on tumor growth or survival. Interestingly, VIP924 was the only ADC to cause a significant increase in peripheral T regulatory cells, which may enhance tumor growth inhibition. Loncastuximab teserine had a deleterious effect on several hematologic parameters, which was not observed with VIP924 or polatuzumab vedotin. The efficacy and tolerability observed in this study with VIP924 suggests further testing in human clinical trials is warranted.

1775. Predictor Factors Of Severe Forms Of Rickettsial Infections

Abstract Background Rickettsial infections usually mimic benign viral infection due to similarities in clinical symptoms. However, severe forms and complications might occur changing therfore the prognosis of the disease. We aimed to study the predictor factor of severe forms of rickettsial infections. Methods We conducted a retrospective study including all patients hospitalized in the infectious diseases department for rickettsiosis. The diagnosis was confirmed by serologies (seroconversion) and/or positive polymerase chain reaction for Rickettsia in skin biopsy. Severe forms included cases with acute respiratory distress syndrome, myocarditis, meningitis, meningoencephalitis, renal failure or septic shock. Results We encountered 472 cases among which 98 cases were severe (20.8%). Median age was 43[25-60] years among severe forms and 38[24-53] years among non-severe forms (p=0.132). Maculopapular skin rash (56.7% vs 88% ; p< 0.001), arthralgia (56.1% vs 77.3% ; p< 0.001) and eschar (16.3% vs 27% ; p=0.029) were significantly less frequent among severe forms. Meningeal syndrome (34.7% vs 8.3% ; p< 0.001), vomiting (56.1% vs 41.8% ; p=0.011) and cough (28.6% vs 13.9% ; p=0.001) were significantly more frequent among severe forms. Leukocytosis was significantly more frequent among severe forms (35.7% vs 20.6% ; p=0.002). The optimal cut-off value of white blood cell count for predicting severe forms was 7595/mm3, with a sensitivity of 61.9% and a specificity of 59.7%. The area under the curve was 0.622 and the 95% confidence interval ranged from 0.558 to 0.686. Elevated C-reactive protein levels (66.3% vs 65.5%; p=0.879) and accelerated erythrocyte sedimentation rate (36.7% vs 40.1%; p=0.543) were noted among severe and non-severe forms of rickettsiosis, with no significant difference. Conclusion The revealing symptoms of severe forms were characterized by the presence of vomiting, cough and meningeal syndrome, in the absence of the typical maculopapular skin rash and eschar. Leukocytosis was more frequent among severe forms of rickettsiosis. The diagnosis of severe forms should be kept in mind in order to promptly initiate the adequate treatment. Disclosures All Authors: No reported disclosures

Corneal Clarity after Phacoemulsification: Nuclear Management by Stop and Chop Method

Introduction: Restoration of effective vision is the principal aim of all modalities of cataract surgery including phacoemulsification. In this method, nuclear part of the cataractous lens is emulsified by ultrasonic power. Energy level of ultrasonic power and turbulence of fluid in the anterior chamber play an important role in corneal endothelial cell loss which inturn influences post-operative corneal clarity. Endothelial cell loss can be minimized by modification of nuclear management method. The common nuclear division techniques of phacoemulsification are stop and chop, divide and conquer and phaco chop. Purpose: To show post-operative corneal clarity after phacoemulsification by stop and chop technique of nuclear divisions. Materials and Methods: This prospective observational study was conducted at selected outdoor patients from January, 2023 to June, 2023 over 33 patients of age related cataract selected for phacoemulsification surgery. Patients were selected based on specific selection criteria. Selected patient underwent detail, ocular and systemic examinations as well as relevant investigations which included assessment of corneal endothelial cell count and central corneal thickness. Phacoemulsification were done in all patients by a single expert surgeon where technique of nuclear management by stop and chop were chosen randomly. Patients were followed up on 1 week, 4 weak and 12 weak after surgery. Corneal endothelial cell count and central corneal thickness were assessed in each visit. All the relevant data were presented by appropriate tables. Results: The mean age of the study subjects were 60.96 with standard deviation 1.75 years out of which 14 were males and 17 were females. Pre-operative value of mean endothelial cell count was 2620+-70.20/mm2. Mean value of endothelial count was 2420+-68.90/ mm2, 2345+-66.64/ mm2, 2310+-66.04/ mm2 respectively after 1 week , 4 weeks and 12 weeks after surgery. Mean value of central corneal thickness was 545+-40.50 micrometer, 535+-40.02 micrometer, 532+-39.85 micrometer respectively after 1 week, 4 weeks and 12 weeks after surgery. Conclusion: Quantitative assessment of endothelial cell count and central corneal thickness shows that at end of the study there was no significant difference in mean endothelial cell count and mean central corneal thickness . Medicine Today 2023 Vol.35(2): 118-120

PSVIII-20 A Premium Dry Cat Food Enhances Digestibility and Gut Health in Cats Compared with a Grocery Dry Cat Food

Abstract As the awareness of pet owners to the benefits of nutrition on pet health and wellness increases, pet owners are looking for foods that include high-quality ingredients and functional benefits. However, little research is available on cats comparing health measures when feeding premium foods vs basic cat foods. This study aimed to compare the effects of feeding a premium cat kibble diet (Blue Buffalo Tastefuls; BB) and a basic grocery cat kibble diet on digestibility, blood biomarkers, and fecal microbiome in healthy adult cats. Sixty cats (mean BW = 4.2 ± 0.8 kg; mean age = 4.3 ± 3.2 yr) were randomly selected to consume BB (n = 30) or grocery diet (n = 30) for 180 days. On days 0, 90, and 180, fresh fecal samples were collected for fecal microbiome and dysbiosis index analysis and blood samples were collected for measurements of complete blood count, serum chemistry, and serum biomarkers. A separate 10-day digestibility study was conducted using 14 cats (BB n = 7, grocery diet n = 7; mean BW = 3.8 ± 0.7 kg). Total fecal samples were collected from day 6 to day 10 for nutrient digestibility analysis. Protocols were approved by the facility’s Institutional Animal Care and Use Committee before the study. All data were analyzed using the Mixed Models procedure of SAS. Fecal scores were not changed (P > 0.05) by diet. Greater (P < 0.05) digestibility of protein, fat, and energy were observed in cats consuming BB. Although all blood cell count and blood chemistry values were within the reference ranges, measurements such as total protein, aspartate transaminase, alkaline phosphatase, cholesterol, and triglycerides were significantly different (P < 0.05) between diet groups. Greater (P < 0.05) insulin-like growth factor-1 was observed in cats fed BB. Cats consuming BB had a lower (P < 0.05) dysbiosis index (less dysbiotic microbiome) and a greater (P < 0.05) bacterial species richness. Relative abundances of 28 bacterial genera were altered (P < 0.05) by the diets, with greater abundances of Faecalibacterium, Megamonas, and Ruminococcus observed in cats fed BB. The data suggest that a premium dry cat food may elicit beneficial effects on digestibility and the GI microbiome of healthy adult cats.

PSVIII-21 A Premium Dry Dog Food Elicits Gut Health Benefits Compared with a Grocery Dry Dog Food

Abstract With the pet humanization trend, pet parents are continuously seeking approaches to improve the health of their pets. Compared with grocery dog foods produced to meet the basic needs of the dog, premium dog foods include high-quality ingredients and functional benefits that can be more appealing to consumers. However, little research is available comparing health measures in dogs fed premium foods vs. basic grocery foods. The objective of this study was to compare the effects of feeding a premium dog kibble diet (Blue Buffalo; BB) and a grocery dog kibble diet on digestibility, blood biomarkers, and fecal microbiome in healthy adult dogs. Sixty dogs (mean BW = 10.8 ± 2.0 kg; mean age = 8.2 ± 3.3 yr) were randomly selected to consume BB (n = 30) or the grocery diet (n = 30) for 180 days. On days 0, 90, and 180, fresh fecal samples were collected for fecal microbiome and dysbiosis index analysis and blood samples were collected for measurements of complete blood count, serum chemistry, and serum biomarkers. A separate 10-day digestibility study was conducted using 12 dogs (BB n = 6, grocery diet n = 6; mean BW = 12.0 ± 2.1 kg). Total fecal samples were collected from day 6 to day 10 for nutrient digestibility analysis. Protocols were approved by the facility’s Institutional Animal Care and Use Committee before the study. All data were analyzed using the Mixed Models procedure of SAS. Stool quality was not different (P > 0.05) between diet groups. Both diets were highly digestible without dietary effects (P > 0.05). All blood cell count and serum chemistry values were within the reference ranges. However, dogs fed the grocery diet had greater (P < 0.05) liver enzyme concentrations including alanine transaminase, alkaline phosphatase, and gamma-glutamyl transpeptidase, and greater (P < 0.05) monocyte counts compared with BB. Dogs had greater (P < 0.05) serum pancreatic polypeptide when fed BB. A lower dysbiosis index (less dysbiotic microbiome) was observed in dogs consuming BB. Relative abundances of 23 bacterial genera were altered (P < 0.05) by the diets, with greater abundances of Bifidobacterium and Megamonas and decreased abundance of Streptococcus observed in the BB group compared with the grocery diet group. These results suggest that premium dry dog food could potentially elicit a positive impact on blood parameters and GI health.

PSVIII-19 Saccharomyces Cerevisiae Fermentation Product Modulates Blood Cell Counts and Gut Immunity in Healthy Adult Cats

Abstract Dietary Saccharomyces cerevisiae fermentation product (SCFP) has been shown to modulate immune responses in dogs and livestock. Information about feeding SCFP to cats, however, is lacking. This study aimed to evaluate the effects of SCFP (TruMune, Cargill, Inc.) on immune indices including fecal IgA, serum oxidative stress markers, and toll-like receptor (TLR) responsiveness in healthy adult cats. Sixty-three cats (mean age = 6.3 ± 2.8 yr; mean BW = 4.4 ± 0.1 kg) were used in a randomized complete block design. All cats were fed a control kibble diet without SCFP (CON) during a 21-day adaptation period. Cats were randomly assigned to one of the three diets including CON, a dry diet containing 150 mg/kg BW SCFP (TRUL), and a dry diet containing 300 mg/kg BW SCFP (TRUH) and fed for 42 days. Fecal samples were collected on days 0, 21, and 42 for fecal IgA analysis. Serum samples were collected on days 0 and 42 for measurements of serum chemistry, complete blood count, and oxidative stress markers. Whole blood samples were collected on day 42 for TLR responsive evaluation. Protocols were approved by the facility’s Institutional Animal Care and Use committee before the study. Data were analyzed using R v.4.1.2. Differences P < 0.05 were considered significant and P < 0.10 as trends. All serum chemistry and blood cell count values were within the reference ranges, with serum chemistry not affected by SCFP. Absolute cell counts of eosinophils and neutrophils decreased linearly with SCFP dose (P < 0.001) while absolute cell counts of lymphocytes and monocytes increased linearly with SCFP dose (P < 0.01). TLR responsiveness was not altered (P > 0.05) by SCFP. A tendency (P = 0.08) of quadratic treatment dose response was noted for thiobarbituric acids from the thiobarbituric acid reactive substance assay, with cats fed TRUL having the greatest thiobarbituric acid values. Fecal IgA showed a quadratic treatment dose response on day 42 (P = 0.01), with cats fed TRUL having the lowest fecal IgA values. These results demonstrate that SCFP may affect immune outcomes including blood cell counts and fecal IgA in adult healthy cats.

Machine Learning Approach to Understand Real-World Treatment in Patients with Higher-Risk Myelodysplastic Syndromes

Background: Higher-risk myelodysplastic syndromes (HR-MDS) is a severe and aggressive form of MDS that represents a group of rare and heterogeneous hematologic disorders that primarily affect the bone marrow and blood cells. Treatment typically involves a combination of supportive care and active treatment with targeted therapies, hypomethylating agents, chemotherapy, and hematopoietic stem cell transplant. In this study, we have evaluated real-world treatment patterns in patients with HR-MDS with machine-learning (ML) models to identify clinical and demographic features that may impact decisions about which patients receive systemic therapies and when the therapies are implemented. Methods: We identified 821 patients aged ≥18 years with HR-MDS diagnosed between January 2015 and April 2022, using ConcertAI's RWD360 database linked with open claims data. RWD360 consists of structured records from US-based oncology electronic health record systems. Demographic and clinical data up to 1 year prior to the date of HR-MDS diagnosis were used to develop a family of XGBoost-based ML models to investigate drivers of treatment. We created five models: a time-to-therapy model that predicts patient-level hazard ratios and four classification models that predict likelihood of therapy in the first 2, 3, 6, and 12 months, respectively, following HR-MDS diagnosis (therapy yes/no models). Model performance was evaluated using Harrell's concordance index (C-index) and Akaike information criterion (AIC) for the time-to-therapy model and area under curve (AUC) for the therapy yes/no models. Key features included laboratory values, Charlson comorbidity index (CCI), and prescribed medications. To better capture the temporal characteristics of a patient's disease state evolution, clinical features were organized into time windows alongside the variations within them. We used recursive feature elimination to bring the total features used in the model to 50 to enhance the interpretability and compactness. Five-fold cross-validation (data was split 5 ways; for each fold, a different 20% is left out for validation, while 80% of the data was used to train the model) on the full 821-patient cohort was performed to train and evaluate the model. Results: The median age in this patient cohort was 73 years (range = 21-88 years); approximately 60% were male, and 66% patients were treated in community oncology practices. Only 42.5% patients were recorded to ever receive systemic active therapy, and of those, the median time to therapy was 4.2 months (Figure 1). Patients had a median hemoglobin concentration of 9.4 g/dL (range = 4.4-18.9 g/dL), median CCI of 1 (range = 0-10), and median polypharmacy of 10 (computed on patients who had at least 1 concomitant medication (49%); range = 1-93) around time of diagnosis (assessed 1 year prior, and up to 30 days after diagnosis date). Model performance for the time-to-therapy model (C-index = 0.78; AIC = 3314.2) and for the therapy yes/no models (AUC = 0.7-0.74) were evaluated. A key predictive feature across models was the minimum laboratory value of white blood cell (WBC) count near the point of diagnosis; low WBC values around time of diagnosis was consistently a strong predictor for receiving systemic therapy and receiving it earlier. Two other top predictors of systemic therapy and its timing were the rate of fall in measurements of blood glucose and the rate of fall of hemoglobin during the 180 to 30 days before diagnosis. Table 1 details model performance and the top 3 features impacting time and likelihood of therapy for all 5 models. Conclusion: We have built a family of data-driven ML models for patients with HR-MDS, based on data collected in routine clinical practice, towards predicting time-to and probability-of first-line systemic therapy with reasonable accuracy driven by patient characteristics. A majority of patients diagnosed with HR-MDS in this study appeared to receive delayed or no systemic therapy in their treatment journey. To bridge gaps in care and facilitate timely treatment in eligible patients, data-driven predictive modeling that take into account detailed patient-level factors is critical. Further analysis on outcomes of untreated patients linked with their key clinical characteristics identified using these models may provide opportunities to improve clinical care by implementing strategies for treatments based on patient-level factors.

Growth Performance, Feed Utilization, Hematological Parameters, and Histological Features of Nile Tilapia (Oreochromis niloticus) Fed Diets with Supplementary Herbal Extracts Under Prolonged Water Exchange

Abstract Nile tilapia is known for its relative tolerance to some biotic and abiotic stressors. However, long-period water exchange may impair the health status and, thereby, the growth performance and feed utilization. In this regard, using herbal extracts may help to relieve the adverse impacts of low water quality on the productivity of this fish species. A 100-day feeding trial was designed to examine the effects of dietary supplements of Aloe vera leaves (AVE), and chamomile flowers, Matricaria chamomilla (CFE) extracts on water quality, growth performance, and well-being of Nile tilapia. Fish (3.95±0.05 g, n=1200) were distributed into five groups (15 fiberglass tanks, 2 m3) under a water exchange rate of 20% every two days for standard control (T0) without herbal supplements. Groups (T1, T2, T3, and T4) were exposed to a long water exchange period of 50% every month, wherein T1 (stressed control), T2, T3, and T4 groups were fed a diet supplemented with 0% herbal extract, 1% AVE, 1% CFE and 0.5% AVE +0.5% CFE, respectively. Results showed that feeding on a T2 diet exhibited a clear improvement under a long period of water exchange stress, and it is almost similar to their counterparts under normal conditions (T0) in terms of specific growth rate, survival rate, feed intake, feed conversion ratio, and protein efficiency ratio. Moreover, the lowest levels of aspartate aminotransferase were obtained with T2 and T0. In addition, the highest white blood cell count and hemoglobin values were recorded with group T0, followed by T2, and T4, while there was no significant difference between T3 and T1. Fish under stressed conditions without herbal extracts (T1) showed the lowest performance and survival rate compared to T0 and T2 groups. In conclusion, feeding diet supplied with 1% A. vera extract to stressed fish restored their performance and well-being to the level of their counterparts under normal conditions.

An evaluation of the effects of glabridin and dexamethasone in bleomycin-induced pulmonary fibrosis: The role of BKCa channels

Pulmonary fibrosis is a refractory entity with a progressive course and no effective therapeutic options. The purpose of this study was to investigate the potential involvement of both glabridin and dexamethasone (Dex) in inflammatory and fibrotic responses in a bleomycin (BLM)-induced pulmonary fibrosis model. The role of Ca+2-activated K+ channels (BKCa) in the anti-inflammatory effects of glabridin was also examined. Adult female Wistar rats were divided into six groups: saline control, BLM, BLM+Gla (BLM+glabridin), BLM+IbTX+Gla (BLM+iberiotoxin+Gla, BKCa channel blocker), BLM+Dex, and BLM+Veh (BLM+dimethylsulfoxide). Inflammatory cell count values, and interleukin (IL)− 6, tumor necrosis factor (TNF)-α, glutathione (GSH), and malondialdehyde (MDA) levels were measured in bronchoalveolar lavage (BAL) fluid in order to measure fibrosis and the extent of tissue damage, in addition to stereological, immunohistochemical and histopathological examinations. Whole-body plethysmography was used to evaluate pulmonary function. Treatments with glabridin and Dex significantly reduced pathological injury and fibrosis in lung tissue, levels of TNF-α and IL-6 increased by BLM, oxidative stress, and fibrillin-1 scoring. Glabridin and Dex also reversed the increases observed in neutrophil, lymphocyte, and macrophage counts in BAL fluid induced by BLM. Glabridin and Dex were found to ameliorate the abnormal course of PIF, PEF, EV, TV, f, and Penh values caused by BLM. Our findings suggest that glabridin and Dex may exert anti-fibrotic effects by suppressing oxidative stress and inhibiting the inflammatory response, and that glabridin may improve pulmonary function through activation of BKCa channels. Both glabridin and Dex may therefore be of therapeutic use in pulmonary fibrosis.

Beneficial effects of antioxidant micronutrients during peri-parturient period on reproductive, udder and body performance of crossbred cows

BackgroundThe critical and stressful peri‑parturient period in the life of a dairy animal creates challenges for dairy owners to keep the animal in the optimum condition of health and hence production. Micronutrients with antioxidant properties help in reducing oxidative stress. A combination of antioxidant micro-nutrients was investigated in the ration of crossbred cows with a hypothesis to eliminate the negative effects of oxidative stress during the stressful peri‑parturient period. MethodsTwenty healthy advanced pregnant crossbred cows reared under similar conditions of feeding with no history of mastitis during previous lactation were selected randomly for the study. Cows were equally distributed randomly into two groups keeping the parity, body condition score, and last record of lactation comparable. Cows were fed the same basal ration with the inclusion of a combination of antioxidant micronutrients (having trace elements copper and zinc with vitamins A and E as per the standard recommendations for cows for the prevention of mastitis) in the diet of the treated group daily, while another group served as control. ResultsThe overall average total (5 %) and fat-corrected milk yield (10 %) remained high in the treatment group. The fat, protein, and lactose content of milk, as well as milk pH, remained statistically comparable (P > 0.05) between the two groups. The overall mean values of milk solid-not-fat remained significantly high (P = 0.04) with a reduced value of the somatic cell count (P = 0.03) in the treatment group indicating the advantage of the antioxidant micronutrients inclusion in the diet. The incidences of mastitis reported in the treatment group were only 14 % (8 Vs 56) as compared to the control group across the entire study period. Higher (P = 0.023) body condition scores in the treatment group cows during the prepartum period lead to lower (0 Vs 40 %) incidences of peri‑parturient reproductive disorders. ConclusionsIt may be concluded that supplementation of antioxidant micronutrients in the dairy cow's ration helps in preventing the adverse effect of oxidative stress during the stressful transition stage and is hence capable to reduce the incidences of mastitis and peri‑parturient reproductive disorders in addition to an enhancement in solid-not-fat content of the milk.

Comparing Hematological and Biochemical Profiles of Pregnant and Non-pregnant Barb Mares Raised in Tiaret, Algeria

Background: The pure-bred Barb horse is a beloved breed from the Great Maghreb. Despite the breed’s prominence in Algeria, no gestational hematological or biochemical research has been done on this breed. Objectives: This study aimed to compare the hematological and biochemical parameters of pregnant and non-pregnant Barb mares in the first, second, and third trimesters of pregnancy. Methods: From 12 pregnant and 6 non-pregnant mares, 102 venous blood samples were taken, and their glucose (Glu), cholesterol (Cho), triglycerides (TG), total protein (TP), urea (Urea), Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), iron (Fer), calcium (Ca), phosphorus (P), and ferric reducing ability of plasma (FRAP) were assessed as biochemical variables. Also, red blood cells, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration, white blood cells, and platelets were all measured as hematological variables. Results: The levels of ALP, ALT, GGT, and P decreased significantly throughout gestation, while Ca, TG, Fe, and Glu levels increased. AST concentrations decreased in the second and third trimesters, whereas Cho levels increased in the first and second trimesters. Urea levels increased significantly in the third trimester, and FRAP showed significant differences at different stages of pregnancy. Mean corpuscular hemoglobin concentration was significantly lower in the first and second trimesters, and hemoglobin values were significantly lower in the second trimester. The mean value of white blood cell count was slightly higher in late pregnancy, while platelet values significantly increased throughout all trimesters. Conclusion: The study provides valuable information on the changes in hematological and biochemical parameters during pregnancy in Barb mares. These findings can be used as a reference for future studies on the reproductive physiology of this breed.

B-143 Distribution of Uricemia Levels in the Population of a Third-level Hospital. Study of the Prevalence of Hypouricemia

Abstract Hypouricemia is a serum uric acid concentration ≤2.0 mg/dL. The causes may be congenital or acquired. Despite not being recognized as a relevant biomarker, the knowledge of the importance of uric acid in redox homeostasis and the hierarchization of hyperuricemia as an independent risk factor in metabolic diseases justifies a reassessment of the significance of the state of acquired hypouricemia. We aimed to study the prevalence of hypouricemia and its relationship with the referred patients’ ages, genders, and morbidities. A descriptive, quantitative study was carried out, through the survey and analysis of biochemical records and clinical records. We worked with the database of the laboratory’s system between January and October 2019, with the data groups of adult patients (≥16 years) that included serum uric acid dosage. The concomitant biochemical parameters (Urea, creatinine, white blood cell count, glucose) were evaluated, and the diagnoses and treatments were collected in the electronic medical record. To define the prevalence, we worked with the first analysis group of each patient. All statistical analyzes were carried out with the Statistix™ software version 7. The criteria for statistical significance were P < 0.05. We found that of the total 159 971 data groups from January to October 2019, 17 983 were selected. There were 234 patients with hypouricemia analyzed. The prevalence of patients with hypouricemia was 2.22%. In our population, postmenopausal women did not match the distribution of uricemia levels to that of men, nor did the mean values of each distribution. Men had higher levels of uric acid than women in any age situation. The prevalence of hypouricemia agrees with what is described in the literature. Of women with hypouricemia, 39% were in the process of pregnancy. Besides that, most patients had a pathology, less than 10% were present for health control, or did not have data in the clinical record. The prevalence of oncological pathology (46%) as a comorbidity in patients with hypouricemia (oncohaematological tumors more frequent than solid ones) and diabetes (22%) was striking. Hypouricemia values were more frequent in outpatients (70%) than in hospitalized patients, however, the last ones showed lower values (1.46 ± 0.4 mg/dL vs 1.52 ± 0.35 mg/dL). The white blood cell count values (0.1–26 * 103 cells/µL) and glucose (0.3–3.76 g/L) that accompany the hypouricemia situation behold pathological values, and the urea and creatinine values were normal. Because this is a retrospective work, we do not have studies of fractional excretion of uric acid, which could delve into the causal research of hypouricemia. Further studies are needed to elucidate the causes of hypouricemia and to clarify its value in diagnosing and treating diseases.

Phenotypic Classification and Clinical Examination of Anemia in Iraqi Water Buffalo in Nasiriyah Governorate

This research aims to classify anemia cases in the Iraqi water buffalo population by collecting 140 blood samples from buffaloes in Nasiriyah Governorate pastures. These samples were collected randomly from three age groups: > 1 year, 1-4 years, and < 5 years. Blood sample analysis revealed 45 anemia cases (32.12%) and 95 healthy cases (67.88%). Phenotypic classification of anemia encompassed microcytic hypochromic (12.14%), macrocytic hypochromic (7.85%), normocytic hypochromic (6.42%), and normocytic normochromic (5.71%) cases. Erythrocyte sedimentation rate (ESR) notably increased in anemic buffaloes, displaying a statistically significant disparity (P< 0.05) compared to healthy counterparts. Anemia cases exhibited higher neutrophil counts in white blood cell relative differentials. For the three age groups, hemoglobin (Hb), packed blood cell volume (PCV), and total red blood cell count (RBC) values indicated significant decreases from normal levels, showcasing statistical significance (P< 0.05) between healthy and anemic buffaloes. While anemia cases generally displayed normal iron concentrations, microcytic anemia demonstrated lower iron levels in the 1-4 and >5 years age groups, with iron levels reaching the minimum global normal range in the <1 year age group. Copper concentrations remained normal in all healthy cases and anemia cases, except for microcytic anemia, which showed reduced levels across age groups.
 In conclusion, this study comprehensively characterizes anemia in Iraqi water buffaloes through clinical, hematological, and elemental analyses. The findings underscore the prevalence of various anemia types, their age-related variations, and significant hematological deviations in anemic buffaloes compared to healthy counterparts. This research enhances our understanding of anemias' impact on this population and provides valuable insights for future diagnostic and management strategies.

Assessment of Blood Volume in Liposuction Fluids Using Colorimetry.

The possibility of using a color contrast method to evaluate blood loss during liposuction was assessed. A color chart of blood-lipid content associated with different blood volumes was developed. Three color cards with different concentrations of blood were developed based on clinical parameters. The color cards were used to evaluate the volume of blood present in liposuction solutions obtained from 60 clinical liposuction patients. The red blood cell count also was evaluated for each patient. The data obtained using each evaluation method were compared and statistically analyzed to determine the most accurate calculation formula. The red blood cell counts were compared to the color card results. The paired t test results for the calculated values for the 3:1, 2:1, and 1:1 color cards and the red blood cell count values were comparable (44.3 ± 22.1 ml vs. 53.6 ± 25.0 ml, t = 10.5; 45.4 ± 19.0 ml vs. 55.2 ± 20.7 ml, t = 18.1; 41.9 ± 25.6 ml vs. 52.8 ± 28.3 ml, t = 14.0). The P values were < 0.05, and the difference between the two groups was statistically significant. The average standard error of the mean was 0.90, 0.54, and 0.77, respectively. Sixty samples were evaluated in a scatter diagram using the two detection methods. Trend analysis revealed that the two results demonstrated a linear increase (y = 5.6 + 1.1x), R2 = 0.989, indicating that the two inspection methods were highly correlated with only small errors. The colorimetric card protocol developed in this study could quickly, accurately, and conveniently calculate blood volumes in liposuction fluids, which has considerable clinical significance. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors http://www.springer.com/00266 .

Interference of red cell parameters by an endogenous substance: An interesting case report

A 6-month-old infant was brought to the hospital with fever, vomiting, loose stools, and abdominal distension. The venous blood sample was sent to the hematology laboratory for complete blood count (CBC). Analysis of the sample on the automated hematology analyzer showed a discrepancy in red blood cell count, hemoglobin, and hematocrit values. Examination of the blood sample revealed a cherry red color appearance. On centrifugation, the plasma was lactescent. This finding provided an essential clue to the patient’s diagnosis of severe hypertriglyceridemia. Turbidity of lactescent plasma interferes with CBC. Analytical errors in the CBC using an automated hematology analyzer caused by lactescent plasma and methods to resolve the false values will be discussed in this case report.

Dietary Effects of Iron and Iron Oxide Nanoparticles on Growth, Muscle Composition and Hematological Aspects of Labeo Rohita

Two hundred Labeo rohita fingerlings of uniform size (2-3g), irrespective of sex, were used to investigate the effects of dietary Fe and Fe2O3-NPs on growth, muscle composition and hematological aspects of fish. Fish maintained on 32% crude protein, conventional basal feed was supplemented with Fe &amp; Fe2O3 nanoparticles (each at 0.6 &amp; 0.8 mg/kg supplementation levels) to replicate groups of fish for 90 days. Morphometric characteristics i.e. wet body weight (g) and body length (cm) was recorded fortnightly. It was found that the weight gain, body length, SGR and PER were significantly improved in the fish fed on nano iron oxide supplemented diet (T2). Protein and lipid nutrients retention in the body meat was improved in group of fish fed with nano iron supplemented diet (T3). A significant increase in hemoglobin (Hb) content, red blood cell (RBC) count, white blood cell count (WBC) and hematocrit (Ht) value was noticed throughout the study period when compared to control group. From the results, it was concluded that Fe-NPs@0.6mg/Kg level is more suitable for Labeo rohita with great growth potential and improved muscle composition and hematology as compared to other levels of Fe and Fe2O3 NPs.

Hematological indices among young adults with homozygous sickle cell disease in steady state in southwestern Nigeria: a case–control study

Abstract Background: Sickle cell disease is a genetic abnormality involving the haemoglobin synthesis. Although it is a primary red cell disorder, white blood cells (WBC) and platelets are also affected by the mutation. Aim: To assess and compare the hematological parameters of homozygous sickle cell disease in a steady state with age-matched and sex-matched hemoglobin AA controls. Methods: A total of 150 apparently healthy young adults (18–35 years) participated in this study. They were classified into two groups 88 cases with hemoglobin SS and 62 controls. Venous blood collected into EDTA tubes from participants were used for the full blood count analysis which was done using hematological autoanalyzer (SFRI blood cell counter, H18 light, France). The data were analyzed using descriptive and inferential statistics and the alpha value was set at P value less than 0.05. Results: The mean values for white blood cell, mean corpuscular hemoglobin, mean cell volume, red cell distribution width and platelet count were significantly higher in the Hb SS cohort while the packed cell volume and red cell count were significantly lower in the Hb SS cohort. Conclusions: homozygous sickle cell disease patients have lower values of hemoglobin concentration, PCV, the red cell indices but higher values of white cell count and platelets compared with hemoglobin phenotype AA controls.

Complete blood and absolute CD4+ve T lymphocyte counts of whole blood donors at a hospital blood center in Nigeria

Background: Comprehensive routine screening and adequate selection of the prospective whole blood donor protects his health and safety as well as that of the recipient. Objectives: The main objective of this study was to determine the complete blood and absolute CD4+ T lymphocyte cell counts of apparently healthy prospective whole blood donors. Participants and Methods: This was a hospital based prospective study. A form was designed by the researchers for data collection. The socio demographic status, complete blood cells and absolute CD4+vel counts of apparently healthy whole blood donors who had tested seronegative for HIV, hepatitis B and C markers was captured . Obtained data was analysed with the statistical package for the social scientist software version 20 Results: One hundred male (97.1%) and three female (2.9%) apparently healthy prospective whole blood donors were studied. The median age of study subjects was 30 years. Obtained median blood cell values were 13g/dl, 40%, 4.9/nl and 203.9/nl for haemoglobin concentration, haematocrit, total white cell and platelet counts respectively. The median values for the mean corpuscular haemoglobin concentration (MCHC), mean corpuscular haemoglobin (MCH) and mean corpuscular volume (MCV) of studied blood donors were 32.6g/dl,27.7pg and 85.7fl respectively. Observed prevalence of subnormal blood cell counts for haemoglobin concentration, total white cells and platelets were 12.6%, 25.2%, 13.6% respectively. Also subnormal values for MCHC, MCH, and MCV were 11.7%, 26.2%, 16.5% respectively among studied whole blood donors. No higher than normal blood cell count values were observed. Median values for erythrocyte sedimentation rate and CD4+ T lymphocytes were 8.4mm/hr and 876.2cells/µl respectively. The percentage of subnormal CD4+T lymphocyte count was18.4% Conclusion: A significant percentage of apparently healthy prospective whole blood donors had subnormal blood cell and CD+ve T lymphocyte values. Obtained normal values were comparable with local reference range reports from previous studies in Nigeria and other parts of Africa.

Reduction of intramammary infections in dairy cow herd by application of mastitis suppression procedures

The aim of this study was to reduce the prevalence of mastitis by introduction of effective anti-mastitis measures in a herd of 125 dairy cows. The effectiveness of the relevant measures was monitored by six examinations conducted in two-month intervals during the one-year period. A reduction in the prevalence of mastitis was recorded from 53.6% to 22.9%, i.e. by 26.0%. The prevalence of Staphylococcus spp. as the most frequently isolated pathogens of the mammary gland in the examined samples gradually reduced from 33.4% to 18.4%, 14.1%, 10.0%, 7.6%, and 8.1%. In contrast to the dynamics of mastitis, the monthly fluctuations in the values of somatic cell count (SCC) and total bacterial count (TBC) in bulk tank milk samples were irregular. However, a declining dynamics of SCC and TBC was evident during the last three samplings which reflected a reduction in the counts of udder pathogens after the treatment and introduction of mastitis suppression procedures.