RBC Count Research Articles

Evaluation of dietary rosemary leaf powder on growth, carcass composition and hemato-biochemical profiles of common carp (Cyprinus carpio) reared in cage system.

This study was conducted to evaluate the effect of rosemary (Rosmarinus officinalis) leaf powder (RLP) at 0.0% (control), 0.5%, 1% and 2% as a dietary supplement for eight weeks, on the growth performance, carcass composition and haemato-biochemical parameters. One hundred twenty common carp (Cyprinus carpio) (21.11±0.17g) were randomly distributed to 12 cages (0.2 m3) with three replicates for 56 days. The results showed that RLP-supplemented diets achieved significant enhancement of growth performance (FBW, SGR and WG) and feed utilization (FCR, FCE and PI) compared with a control group. In addition, TFI and PI were significantly improved in 0.5 and 2% RLP-supplemented groups. RLP had no significant effects on body composition except moisture content which was significantly increased in fish fed with a 1% supplemented diet and crude protein which was significantly decreased in fish fed with a 1% supplemented diet compared to the control diet. The results showed a significant increase in RLP-supplemented diets in RBC counts, LYM, MCV, hematocrit, total protein, albumin and globulin compared with the control group. WBC counts significantly increased in fish fed the 0.5 and 2% RLP diets, and hemoglobin significantly elevated in fish fed the 1% RLP diet. Glucose, GRA, MCH, MCHC and MPV were significantly decreased in the group fed the RLP diets compared with the control group. The AST, ALT, LDL, VLDL, CHO and TG levels showed a significant decrease in RLP compared to the control diet (P < 0.05), while ALP level fluctuated (P > 0.05). The urea and creatinine levels were higher in the RLP groups compared with the control (P < 0.05). Dietary RLP 1% increased lipase when compared to the control group. However, amylase levels in fish fed 0.5 and 1% RLP has significantly improved, while fish fed 2% RLP has significantly decreased when compared to the control diet. Overall, the results demonstrated that dietary RLP supplementation enhanced the growth and health status of common carp.

Comparable Evaluation of Nutritional Benefits of Lactobacillus plantarum and Bacillus toyonensis Probiotic Supplementation on Growth, Feed Utilization, Health, and Fecal Microbiota in Pre-Weaning Male Calves.

This study was conducted to investigate the impact of probiotic supplementation using Lactobacillus plantarum DSA 20174 and/or Bacillus toyonensis ATCC 55050 on growth performance, blood parameters, hematological measures, and fecal microbiota in pre-weaning Holstein calves. Thirty-two four-day-old male calves with a similar genetic background, weighing an average of 38.27 ± 0.12 kg, were randomly assigned to four groups. The groups consisted of a control group (CON) without supplementation, a group receiving B. toyonensis (BT) at 3 × 109 cfu/calf/day, a group receiving L. plantarum (LP) at 1 × 1010 cfu/calf/day, and a group receiving a combination of LP and BT (LP + BT) at half the dosage for each. The study found that calves supplemented with LP and LP + BT experienced significant improvements in average daily gain and final body weight compared to the control group. The LP + BT group showed the most positive effects on TDMI, starter intake, and CP intake. RBC counts tended to be higher in the probiotic groups, with the LP + BT group having the highest values. The LP + BT group also had higher total protein, albumin, globulin, and hematocrit concentrations. All probiotic groups showed higher serum IgG concentrations. Probiotic supplementation led to increased total bacterial count and decreased levels of E. coli, salmonella, and clostridium. The LP + BT group had a significant decrease in coliform count, while both LP and LP + BT groups had increased Lactobacillus populations. In conclusion, LP + BT probiotic supplement showed the most beneficial effects on growth, feed efficiency, blood constituents, and modulation of fecal microbiota composition.

Genetic Deletion or Pharmacologic Inhibition of PTPN11 Impedes the Development and Progression of Myeloproliferative Neoplasms Induced By JAK2V617F and MPLW515L Mutants

Myeloproliferative neoplasms (MPN) including polycythemia vera (PV), essential thrombocythemia (ET) and myelofibrosis (MF) are clonal myeloid malignancies derived from mutated hematopoietic stem cells. The JAK2V617F mutation was found in ~95% cases of PV and 50-60% patients with ET and MF. Mutations in MPL and CALR were also detected in ET and MF. The JAK inhibitors, Ruxolitinib and Fedratinib, can reduce splenomegaly and alleviate constitutional symptoms but they are not sufficient to induce remission of MPN. So, there is an unmet need to identify new therapeutic targets and therapies for MPN/MF. The non-receptor protein tyrosine phosphatase 11 (PTPN11), which positively regulates hematopoietic cell signaling, is constitutively hyperphosphorylated in MPN patient cells and in hematopoietic cell lines expressing MPN driver mutants. So, we hypothesize that PTPN11 plays an important role in MPNs. To examine the role of PTPN11 in JAK2V617F-induced MPN, we generated inducible PTPN11-deficient heterozygous JAK2V617F mice by crossing PTPN11 floxed mouse with Mx1Cre and JAK2V617F knock-in mice. While expression of heterozygous JAK2V617F (JAK2 VF/+) induced a PV-like MPN characterized by increased WBC, neutrophil, RBC, hemoglobin and platelet counts in the peripheral blood and enlargement of spleen size, deletion of PTPN11 normalized the blood counts and spleen size in JAK2 VF/+ mice. PTPN11 deletion significantly reduced the expansion of hematopoietic stem/progenitors, erythroid, megakaryocytic, and granulocytic cells in the bone marrow (BM) and spleens of JAK2 VF/+ mice. Furthermore, deletion of PTPN11 abrogated Epo-independent CFU-colonies, a hallmark feature of PV disease, in the BM and spleens of JAK2 VF/+ mice. We also investigated the role of PTPN11 in the development of MF using homozygous JAK2V617F and MPLW515L mouse models. We found that deletion of PTPN11 significantly reduced WBC, neutrophil and platelet counts, normalized spleen size and abrogated BM fibrosis in JAK2V617F and MPLW515L mouse models. These results suggest that PTPN11 plays an important role in the development and progression of PV and MF. Data from our PTPN11 genetic deletion study led us to investigate the effects of PTPN11 inhibition against MPN/MF. We tested the efficacy of an allosteric PTPN11 inhibitor SHP099 in MPN hematopoietic cell lines and mouse models of MPN/MF. We observed that treatment of SHP099 significantly reduced proliferation of hematopoietic cells expressing JAK2V617F and MPLW515L. SHP099 treatment also significantly inhibited hematopoietic progenitor colony growth in PV and MF patient CD34+ cells. Next, we tested the in vivo efficacy of SHP099 against PV using heterozygous JAK2V617F mice. Treatment of SHP099 significantly inhibited the increase in WBC, neutrophil and RBC counts and markedly reduced splenomegaly in JAK2 VF/+ mice. SHP099 treatment also significantly reduced hematopoietic stem/progenitors and myeloid precursors in the BM and spleens of JAK2 VF/+ mice. Furthermore, SHP099 treatment abrogated Epo-independent CFU-E colony formation in the BM and spleens of JAK2 VF/+ mice. We also tested the efficacy of SHP099 alone or in combination with ruxolitinib against myelofibrosis using homozygous JAK2V617F (JAK2 VF/VF) and MPLW515L mouse models. SHP099 treatment alone significantly reduced WBC and neutrophil counts and markedly reduced splenomegaly in JAK2 VF/VF and MPLW515L mice. Combined treatment of SHP099 and ruxolitinib resulted in greater reduction of blood cell counts and spleen size in JAK2 VF/VF and MPLW515Lmice. Histopathologic analyses showed that SHP099 treatment significantly reduced BM fibrosis in JAK2 VF/VF and MPLW515L mice while combined treatment of SHP099and ruxolitinib resulted in greater inhibition of BM fibrosis in these mice. To gain insights into the mechanism of inhibition of MPN/MF by PTPN11 deletion or inhibition, we performed RNA-seq analysis on purified LSK cells from JAK2 VF/+ and PTPN11deleted-JAK2 VF/+ mice as well as from JAK2 VF/VF mice treated with ruxolitinib and SHP099/ruxolitinib. RNA-seq data analysis revealed that genes related to MYC targets, ribosome biogenesis and translation were significantly down-regulated by PTPN11 deletion or SHP099/ruxolitinib combination treatment. Overall, our results suggest that inhibition of PTPN11 alone or in combination with JAK2 inhibition might be useful for treatment of PV and MF.

Inducible Knockout of Codanin-1: An Adult Mouse Model of Congenital Dyserythropoietic Anemia Type-I

Congenital dyserythropoietic anemias (CDAs) are a heterogenous group of congenital blood diseases, characterized by ineffective erythropoiesis, hemolysis, and the presence of bi- or multi-nucleated erythroblasts in the bone marrow (BM). Several CDA types exist, originally classified by bone marrow pathology and now more accurately by their causative mutations, when known. Mutations in codanin-1 ( Cdan1) are the major contributor (≈80%) to CDA type-I (CDA-I). Patients with CDA-I frequently require transfusions in utero to survive and present with varying degrees of hemolytic anemia with relative reticulocytopenia suggesting ineffective erythropoiesis, binucleated erythroblasts, and, under transmission electron microscopy (TEM), a “swiss cheese” appearance in the nuclear heterochromatin of erythroblasts. Treatment for patients with CDA-I includes blood transfusions and iron chelation, or IFN-α, which allows most treated patients to achieve transfusion independence. However, the molecular mechanisms of dyserythropoiesis in CDA-I as well as those of the therapeutic effect of IFN-α are unknown. To study the pathogenesis of the disease and the mechanisms of IFN-α action in vivo, we generated an adult mouse model of CDA-I. Previous efforts to produce a CDA-I mouse model were achieved via a constitutional, EpoR Cre+/--driven erythroid-specific deletion of Cdan1; however, this mouse suffered embryonic lethality by E12.5 [Noy-Lotan et al, Frontiers in Physiology, 2021]. We developed a transgenic mouse model in which the most common CDAN1 missense mutation (p.R1042W in humans corresponding to mouse p.R1047W) was introduced using CRISPR-Cas9 editing, and the mice were bred to homozygosity. These mice survived to term but exhibited no significant anemia, altered erythropoiesis, or the characteristic “swiss cheese” appearance in the heterochromatin of the erythroblasts under TEM. They demonstrated no differences from control mice in recovery after induction of stress erythropoiesis by phlebotomy (in agreement to the findings reported in parallel by Noy-Lotan et al, 2021), suggesting that this model is inadequate for studying the CDA-I phenotype. Therefore, we proceeded to generate an inducible Cdan1 knockout mouse model to test the hypothesis that deletion of Cdan1 in the erythroid lineage,mediated by Gata1 creERT2 [Yu et al. Blood. 2021], will recapitulate the CDA-I phenotype in adult mice. This maneuver avoids embryonal lethality during the extreme stress erythropoiesis phase of fetal development. We bred mice with the wildtype (WT) or floxed Cdan1 gene ( Cdan1WT/WT and Cdan1Fl/Fl, respectively) with mice carrying Gata1 creERT2. Cdan1WT/WT; Gata1creERT2+/- mice served as WT controls and Cdan1Fl/Fl; Gata1creERT2+/- mice served as the experimental mice with erythroid-specific Cdan1 deficiency after 10 injections of tamoxifen every other day to activate Cre expression. We evaluated the phenotype of these mice using complete blood counts (CBCs), light microscopy, TEM, and flow cytometry of erythroid progenitors and precursors. The Cdan1-deficient mice became anemic with reduction in RBC count, hemoglobin ( Fig. 1), and hematocrit. Light microscopy of the BM and spleen revealed erythroblasts joined by a thin-interchromatin bridge and binucleated erythroblasts in the Cdan1-deficientcompared to WT mice (z-test of proportions, P &amp;lt; 0.001), appearing at a frequency of ≈13% of all erythroblasts (see Fig. 2). TEM revealed a “swiss cheese” appearance in the heterochromatin of erythroblasts, the pathognomonic finding of CDA-I. Flow cytometry of the BM and spleen revealed decreased late-stage erythroblasts, demonstrating ineffective erythropoiesis. To determine a possible stage in which IFN-α may be directly acting on erythropoiesis, we examined the expression of IFNαR1 in erythroblasts. In all mice, basophilic, polychromatic, and orthochromatic erythroblasts had relatively low expression of IFNαR1. Interestingly, proerythroblasts had relatively high expression of IFNαR1 but only in the spleen. These data provide a starting point for localizing the effect of IFNα in the CDA-I phenotype. In conclusion, erythroid-specific deletion of Cdan1 via Gata1 creERT2 in adult mice recapitulates the CDA-I phenotype and provides a viable mouse model to investigate the disease mechanism and the therapeutic effect of IFN-α on CDA-I.

Improved Quality of Life upon Shifting from Darbepoetin to Luspatercept in Patients with Low-Risk Myelodysplastic Syndrome (MDS)

Introduction: Low-risk MDS is characterized by refractory anemia due to Dyserythropoiesis 1. Standard of care treatment is mainly Erythropoiesis stimulating agents (ESA) or blood transfusion or both 1. Response to ESA is achieved in some patients whose erythropoietin (EPO) level is less than 500 mu/ml 2. Luspatercept is a red cell maturing agent that may lead to an increase in Hb of up to 3 g/dL 3 in responders. Response to ESA can be enhanced with the addition of Luspatercept 4. Hereby we are reporting successful shifting of two patients from Darbepoietin to Luspatercept with significant increase in Hb level and improvement in quality of life. Case 1: A 62-year-old man without previous illness presented in April 2018 with progressive symptomatic anemia. His body weight was 107 kg, and his physical examination was unremarkable other than pallor and tachycardia. The laboratory investigation revealed WBC, 5.62 x 10 3/uL, RBC 2.2 x 10 3/uL, Hb 7.8 g/dL, MCV 110.4 fl, platelets 706 x 10 3/uL, and reticulocytes 1%. Blood film showed reduced RBC count with numerous macrocytes and no other abnormalities. Bone marrow revealed 80% cellularity with Dyserythropoiesis, 16% ring sideroblasts, and 2% blasts. The Ferritin level, B12 levels were elevated (table). EPO level was elevated but less than 500 mlU/ml and the cytogenetic analysis showed normal karyotype. However his molecular profiling demonstrated SF3B1 and TET2 mutations. At the time of diagnosis, the Revised International Prognostic Scoring System (IPSS-R) was 2.0, indicating low risk MDS. He was given darbepoetin 300 mcg SC weekly for four weeks without any improvement, then darbepoetin dose was escalated to 500 mcg weekly with a good response. Hb level remained at 8.5 - 9.5 g/dl for three years. The patient's physical performance improved moderately, with an increase in walking distance on flat surfaces from 20 to 35 minutes. In August 2022, he was switched to Luspatercept 100 mcg SC on a three-weekly schedule. Within three months, his hemoglobin level increased to 10.8 g/dl and he experienced significant improvement in physical performance and mental well-being, along with a significant reduction in fatigue, enabling him to increase his walking distance to 76 minutes. Case 2: A 77-year-old man with type 2 diabetes mellitus and mild renal impairment who developed an unexplained progressive anemia. His body weight was 79 kg, and his physical examination was unremarkable other than pallor. His laboratory investigations showed WBC 4.5 x 10 3/uL, RBC 1.5 x 10 6/uL, Hb 5.2 g/dL, MCV 96 fl, platelets 706 x 10 3/uL, and reticulocytes 0.8 %. Blood film showed reduced RBC count, normocytic anemia, and no other abnormalities. Bone marrow revealed 60% cellularity with Dyserythropoiesis, 17% ring sideroblasts, and 1% blasts. His baseline serum creatinine was 1.8 mg/dl, and HbA1C 10.0. Two units of packed red blood cells were transfused every 2- 4 week to keep his Hb level around 8 g/dL. Darbepoetin 80 mcg SC weekly for four weeks,150 mcg weekly for 4 weeks and 300 mcg weekly for further 4 weeks without any response. Eventually, he responded to 500 mcg of Darbepoetin per week. As a result, his Hb increased to the range of 8.6-9.4 g/dl for one year, with moderate improvements in performance status. Darbepoetin was switched to Luspatercept 75 mcg three-weekly in January 2023, resulting in an increase in his hemoglobin level to 10.3 g/dl, with significant improvement in his energy level and overall well-being. Discussion: Blood transfusion and ESA have traditionally been used as standard treatments for low-risk MDS. Recently, Luspatercept in combination with ESA has shown enhanced effectiveness in elevating Hb levels. Both of our patients had significant increases in Hb levels along with a noticeable improvement in quality of life because of Luspatercept alone. Conclusion: Following the switch from Darbepoetin to Luspatercept, there was a significant improvement in Hb level and in quality of life in our 2 patients. This finding needs to be confirmed with larger studies.

Evaluation of biochemical parameters in operating room staff exposed to radiation and anesthetic gases.

Exposure to ionizing radiations and other hazardous agents such as anesthetic gases pose serious risks to the health of healthcare workers. This study aimed to evaluate the changes in blood and biochemical parameters of the operating room staff exposed to ionizing radiations and remnants of anesthetic gas. This cross-sectional study was performed at (Ayatollah Taleghani Hospital). The control group was selected from different parts of the hospital that were not exposed to ionizing radiations and anesthetics, including the office, services, and treatment. The case group included all operating room personnel. Hematopoietic parameters such as complete blood count and WBC differential, and parameters of liver function such as serum activity of liver enzymes (ALT, AST, ALP, LDH) and serum bilirubin levels, fasting blood sugar, serum lipid profile, level of vitamin D and magnesium were measured for the exposed and referent subjects. Additionally, a checklist was used to gather data regarding the occupational variables and medical histories of the studied subjects. The mean values of Hb, Hct, Vitamin D, and MCHC, as well as the RBC count, were significantly lower in the exposed individuals than in the referent subjects. In contrast, the proportion of smokers was significantly higher in the exposed group than in the referent group. No significant differences were noted between exposed and unexposed groups as far as other parameters were concerned. However, no significant differences were noted between the case and control groups as far as other measured parameters were concerned. Likewise, no significant differences were noted between exposed and referent groups as far as blood types, history of underlying diseases, work history, working hours per month, number of morning and evening shift hours, type of diet, consumption of a high-fat diet a day before blood sampling, X-ray in the recent year, history of radiotherapy, and therapeutic agents use was concerned. Exposure of operating room staff to ionizing radiations and waste anesthetics gases is associated with subtle, subclinical prepathologic decreases in some hematopoietic parameters such as hemoglobin, hematocrit and MCHC levels, RBC count as well as vitamin D levels.

Evaluations of haematological profile in geriatric patients in a tertiary care hospital

Ageing is the process which has an impact on all the cells, tissues, organ systems, and organisms. Alterations or modifications in the bone marrow are observed with ageing. Increased risk of myeloproliferative disorders, anaemia, and a deteriorating adaptive immunity are signs of age-related hematologic alterations, as is a loss in bone marrow cellularity. Evaluations of haematological parameters can be used to track the disease's development. Hence, this retrospective study was designed to evaluate the haematological profile of geriatric patients and to compare the haematological parameters between males and females. The Complete Blood Count reports of 445 patients aged 60 years and above were selected for this retrospective study. The backbone computerized data of these cases were taken from central laboratory of Yenepoya Medical College Hospital. Haematological profile was studied and analyzed from selected cases. The independent sample t test was used to compare the haematological parameters of males and females.P value&lt;0.05 was considered as significant.The results obtained showed that there was a statistical significance in lymphocytes, Eosinophils, MCH, MCHC in Geriatrics patients. There was no statistical significance in haemoglobin, neutrophils, monocytes, platelet count, MCV, PCV, and RBC count .Major abnormalities were found in Haemoglobin and lymphocytes than other haematological parameters. Incidence of anaemia was 77.8% and lymphocytopenia 77.9%.The most frequently observed haematological abnormalities in elderly persons were anaemia and lymphocytopenia. Anaemia was seen to be higher in females than males. Lymphocytopenia was seen to be higher in males than females. Early evaluations of haematological profile will reduce morbidity and mortality among the geriatric patients.

Modulatory Effects of Lawsonia inermis, Waltheria indica, Moringa oleifera and Nigella sativa on Haematology, Blood Pressure and Electrocardiographic Parameters of Wistar Rat Exposed to Acute Pain

Background: Pain is an obnoxious stimulus usually triggered by stressors that can affect homeostasis. The inability to sense pain is usually connected to abnormal fluctuations in normal body functions, leading to shortened life expectancy. Purpose: This study aimed to assess the effects of Lawsonia inermis, Waltheria indica, Moringa oleifera and Nigella sativa on hematology, electrocardiography, and blood pressure in Wistar rats experimentally exposed to acute pain. Methods:21 Adult female rats were divided into (1-7) of three rats each. Group 1 (negative control), group 2 (positive control), group 3 (Lawsonia inermis at 200 mg/kg), group 4 (Waltheria indica at 200 mg/kg), group 5 (Nigella sativa at 200 mg/kg), group 6 (Moringa oleifera at 200 mg/kg), and group 7 (Diclofenac at 10 mg/kg). Rats were dosed for 14 days, after which pain was induced. Result: The extracts showed non-significant increases in most hematological parameters, including PCV, RBC, and WBC counts, compared to positive controls. The effects of the extracts on the electrocardiographic parameters were not significant. Although the extract relieved pain, the effect of acetic acid on the heart was not completely ameliorated. The results of this study showed high blood pressure in untreated rats, which could be linked to pain induction. M. oleifera and W. indica exhibited minimal blood pressure-lowering effects. W. indica. and L. inermis showed transient blood pressure-lowering effects. Conclusion: This study concluded that the extract had a modulatory effect on hematology and a lowering effect on blood pressure and electrocardiac parameters in all treatments.

Evaluation of Prevalence and Severity of Anemia among Non-pregnant women of Reproductive Age in Garmian Province, Iraqi Kurdistan Region

Anemia is a major significant public health issue that affects both pregnant and non-pregnant women throughout the world. This study aimed to determine the prevalence and severity of anemia, as well as its classification, based on MCV values in non-pregnant women residing in the Garmian province in the east of Iraq. Blood samples were collected from non-pregnant women between the ages of 16 and 49 years. Anemia is classified as mild, moderate and severe, and as microcytic, normocytic and macrocytic depending on the level of Hb or mean corpuscular volume (MCV) respectively. In this study sociodemographic and hematological characteristics were also evaluated. The results showed that 56% of the study samples were anemic, and of the total anemic patients, 54 (35%), 86 (55%) and 16 (10%) had mild, moderate, and severe anemia respectively. Also, of the total anemic patients, 102 (65%) had microcytic type of anemia, 51 (33%) had normocytic anemia and only 3 (2%) had macrocytic anemia. The majority of anemic women (77.6%) had normal RBC counts but low Hct, MCH and MCHC values of 76.3%, 79.5% and 81.4%, respectively. The results showed also that there was no significant difference between the severity of anemia and age groups, occupation, blood pressure, bleeding, and haemorrhoids. However, diabetics showed a significant difference in anemia severity with &lt;0.0004 P-value. The results showed that anemia had become a major public health issue in the study area.

Diagnostic Value of Red Cell Distribution Width and Red Blood Cell Distribution Width Index in Differentiating between Iron Deficiency Anemia and Beta Thalassemia Trait

Objective: To determine the diagnostic value of red blood cell distribution width and red cell distribution width index in differentiating iron deficiency anaemia and beta-thalassemia trait. Study Design: Cross-sectional study. Place and Duration of Study: Pak Emirates Military Hospital, Rawalpindi Pakistan, from Dec 2021 to Feb 2022. Methodology: A study population of 110 children with an age limit of 1.5-7 years who reported low haemoglobin wassubjected to serum iron and ferritin levels with further evaluation of red cell count, blood morphology, and comparison of mean corpuscular volume. Red cell distribution width was taken from the lab report, whereas Red blood cell distribution width index value was computed for the samples. Results: A total of 110 patients were enrolled in the study. Seventy-five (68.18%) participants were diagnosed with IDA, 26(23.63%) participants were the candidate of βTT. Nine participants were suffering from anaemia of variable etiologies such as chronic ailment and sideroblastic, etc. βTT patients presented with higher RBC counts with a mean of 5.71±1.2 x1012 /l (6.8- 8.5 x1012 /l) when compared to IDA participants who deciphered values of 3.5±0.5 (2.6-4.3 x1012 /l). Mean MCV in βTT was 52.2±2.7(50.1-56.8) whereas for IDA were 72.5±1.2(61.2–78.3). In IDA low serum ferritin was recorded 4.02±1.1(2.2-9.2) as compared to mean values of βTT patients. Conclusion: Red cell distribution width index and Red blood cell distribution width are authentic variables and inexpensive modalities in differentiating between iron deficiency anaemia and beta-thalassemia trait with reliability.

Gallic Acid Attenuates Cardiovascular and Hematological Complications in Vincristine Treatment via Hydroxyl Radical Scavenging and Endogenous Antioxidant Stimulation in Male Wistar Rats

The menace of cancer and the nightmare of complications of cancer chemotherapy have driven researchers to explore simple but efficient combination therapy that includes antioxidants, in cancer therapy. The ability of gallic acid to correct the toxic complication of Vincristine was investigated. Twenty adult male rats of the Wistar strain were grouped into four, randomly, consisting of five rats each. The untreated control (group A) was given only distilled water, groups B and C 0.025 mg/kg Vincristine sulfate intraperitoneally once a week for two weeks. Group C rats were thereafter administered 100 mg/kg gallic acid daily by gastric gavage for 14 days. At 14 days, blood pressure and ECG were measured in the rats, then blood samples were obtained via the retrorbital venous plexus for determination of haematological parameters and plasma biochemistry. They were then euthanized through cervical dislocation, under ether anaesthesia, and liver, kidneys, heart, and brain samples were collected, weighed, and stored for determination of marker of oxidative stress in the post mitochondrial fractions of each organ. Results of the study showed that rats in group B had hypertension as evidenced by elevated diastolic and systolic as well as mean arterial pressure while QT interval and corrected QT were slightly elongated. They also had lowered RBC, WBC, and granulocyte counts. Markers of oxidative stress, GSH, and SOD were also depleted while H2O2 generation increased in this group, whereas all the observed anomalies were corrected in the group C rats that were administered both Vincristine and gallic acid. This study further showed that Vincristine, at normal recommended therapeutic dosage is toxic, causing anaemia, panleucopenia, and cardiovascular anomalies via oxidative stress and generation of hydroxyl radicals. These were however corrected by concurrent administration of gallic acid

Hybrid total arch replacement via ministernotomy for Stanford type A aortic dissection.

Type A aortic dissection (TAAD) is a cardiovascular emergency condition with high mortality rate. Hybrid total aortic arch replacement using endovascular graft for the descending aorta repair results in favorable outcomes and has been recommended as an alternative procedure for the higher-risk category patients. Our institution started applying the upper ministernotomy incision technique for the hybrid procedures back in 2018. We collected patients who underwent hybrid total arch replacement (HTAR) via ministernotomy (96) and total arch replacement with frozen elephant trunk (TAR + FET) procedures (99), between 2018 and 2021. The baseline information, intraoperative and postoperative characteristics have been compared. Kaplan-Meier analysis was used for survival evaluation. Cox regression were applied to identify the independent predictor of mortality. The baseline characteristics between the two patient groups were compared and found similar, except that RBC counts were higher (p = 0.038) and the ascending aorta diameter was smaller (P = 0.019) in the "HTAR" group relative to the "TAR + FET" group. The cardiopulmonary bypass time (P < 0.001), the aortic cross clamp time (P < 0.001), the operation duration (P = .029), ICU (P = 0.037) and postoperative hospital stay (P = 0.002) were shorter in the "HTAR" group. The "HTAR" group exhibited also significantly lower levels of intraoperative transfusion (all <0.001) characteristics than the "TAR + FET" group. The hospital mortality and 1-year mortality revealed similar patterns in both groups. HTAR via ministernotomy have similar short term prognosis, and also reduced the ICU and postoperative hospital stay. In all, The application of the ministernotomy technique in HTAR was safe and technically feasible and may benefit individual patients as well as hospitals in general.

INFLUENCE OF SEASON, ENVIRONMENT, AND SEX ON SELECTED BLOOD PARAMETERS OF GEOFFROY'S SIDE-NECKED TURTLES, PHRYNOPS GEOFFROANUS (SCHWEIGGER, 1812).

Laboratory tests help to determine a diagnosis, to plan treatment, and to indicate prognosis of diseases. However, the interpretation of test results in reptiles is challenging, because they are influenced by environmental and individual factors. Therefore, the objective of this study was to establish hematologic parameters of Geoffroy's side-necked turtle (Phrynops geoffroanus), including variables such as season, environment, sex, and straight carapace length (SCL). Blood samples were analyzed from 38 P. geoffroanus (23 males and 15 females) collected during the rainy and dry seasons. Some of the animals were from captivity and others from an urban stream. To verify the influence of the variables on blood parameters, the general linear model was performed. The Pearson correlation coefficient was also used to verify the association between SCL and hematologic parameters. Among these parameters, hemoglobin (P = 0.008) and mean corpuscular hemoglobin concentration (P = 0.001) were statistically higher during the rainy season. On the other hand, WBC (P = 0.011), heterophils (P = 0.045), eosinophils (P < 0.001), lymphocytes (P = 0.014), and monocytes (P = 0.039) were higher in the dry season. The RBC count (P = 0.035), PCV (P = 0.029), basophils (P = 0.013), and monocytes (P = 0,013) were significantly higher in individuals from captivity, but lymphocytes (P = 0.033) were higher in the urban turtles. Only eosinophils (P = 0.025) were higher in females than in males. There was no influence of the variables season, environment, and sex on the mean corpuscular volume (P = 0.071; P = 0.458; P = 0.052), total solids (P = 0.773; P = 0.121; P = 0.131), and heterophil:lymphocyte ratio (P = 0.992; P = 0.58; P = 0.119). No influence of the size on hematologic parameters was observed. Season, environment, and sex may influence the blood parameters of P. geoffroanus, and these factors should be routinely considered in the interpretation of laboratory results.

Paediatric COVID-19 Outcomes: Haematology Parameters, Mortality Rates, and Hospitalization Duration.

The global COVID-19 pandemic has strained healthcare systems around the globe, necessitating extensive research into the variables that affect patient outcomes. This study examines the relationships between key haematology parameters, duration of hospital stay (LOS), and mortality rates in COVID-19 cases in paediatric patients. Researchers analyse relationships between independent variables (COVID-19 status, age, sex) and dependent variables (mortality, LOS, coagulation parameters, WBC count, RBC parameters) using multivariate regression models. Although the R-square values (0.6-3.7%) indicate limited explanatory power, coefficients with statistical significance establish the impact of independent variables on outcomes. Age emerges as a crucial predictor of mortality; the mortality rate decreases by 1.768% per age group. Both COVID-19 status and age have an inverse relationship with length of stay, emphasising the milder hospitalisation of children. Platelet counts decline with age and male gender, potentially revealing the influence of COVID-19 on haematological markers. There are significant correlations between COVID-19 status, age, gender and coagulation measures. Lower prothrombin time and D-dimer concentrations in elder COVID-19 patients are indicative of distinct coagulation profiles. WBC and RBC parameters exhibit correlations with variables: COVID-19-positive patients have lower WBC counts, whereas male COVID-19-positive patients have higher RBC counts. In addition, correlations exist between independent variables and the red cell distribution width, mean corpuscular volume, and mean corpuscular haemoglobin. However, there is no correlation between mean corpuscular haemoglobin concentration and outcomes, indicating complex interactions between haematological markers and outcomes. In essence, this study underlines the importance of age in COVID-19 mortality, provides novel insights into platelet counts, and emphasises the complexity of the relationships between haematological parameters and disease outcomes.

A-135 Improving Cell Count Turnaround Times With the GloCyte® Automated Cell Counter for CSF: A Retrospective Study at MercyOne Des Moines Medical Center

Abstract Background Accurate, STAT cerebrospinal fluid (CSF) cell counts are critical in the diagnosis of meningitis, malignancy, demyelinating disease, and hemorrhage. Due to historical limitations with the linearity of automated analyzers, CSF cell counts have traditionally been performed manually with a hemocytometer and the aid of a brightfield optical microscope. Manual cell counting is time-consuming, subjective due to reliance on human interpretation, and requires a high degree of skill. This poses significant challenges for clinical laboratories of all sizes facing unprecedented med tech shortages. The purpose of this study was to evaluate the impact of implementing the GloCyte Automated Cell Counter for CSF, an FDA-cleared device with linearity down to 0 cells/µL, on result turnaround times. Methods Result turnaround time data for CSF specimens processed in the hematology lab at MercyOne Des Moines Medical Center, a 600-bed hospital in Des Moines, Iowa, was analyzed in a retrospective study of two cell counting methods- (1) manual cell counting using a Neubauer-Improved hemocytometer and (2) GloCyte, a benchtop cell counting device that uses fluorescence technology and highly specific reagents to automate RBC and TNC cell counts in less than 5 min. Turnaround times for 227 CSF specimens counted manually during an approximately six-month period prior to the implementation of GloCyte (January 2, 2017 to June 29, 2017) were compared to those for 133 specimens analyzed on GloCyte over a similar duration (January 4, 2021 to June 30, 2021). In this study, turnaround times were defined as the time between specimen arrival in the lab and result verification. Correlation between the methods was previously established in the MercyOne Des Moines Medical Center laboratory. Results The data demonstrated a significant, 16–30-minute improvement in turnaround times with GloCyte compared to the manual hemocytometer method. Median turnaround times, analyzed to adjust for add-ons that resulted in longer turnaround times pre- and post-GloCyte implementation, were 1:38h for specimens counted manually vs 1:08 h for specimens analyzed on GloCyte. The average turnaround time for the 227 CSF specimens counted manually was 2:03 h compared to 1:46 h for the 133 samples analyzed on GloCyte. The MercyOne Des Moines Medical Center laboratory processed 456 CSF cell counts in 2022, equating to a reduction in turnaround time of 122–228 h annually with the implementation of GloCyte. Conclusion This study demonstrates that GloCyte can be implemented to significantly improve turnaround times for CSF cell counts. The MercyOne Des Moines Medical Center laboratory realized several benefits from the reduction in turnaround times- GloCyte has (1) freed up significant tech time for other tasks, (2) standardized CSF cell counting to mitigate tech to tech variability, (3) increased lab staff satisfaction and decreased stress levels, and (3) increased physician satisfaction. For MercyOne Des Moines Medical Center’s sister site, MercyOne West Des Moines Medical Center, a 146-bed hospital that infrequently performs CSF cell counts, GloCyte has provided peace of mind in performing cell counts accurately with techs of all skill levels.

B-010 Comparison of Manual Urine Dipstick and Microscopy With Automated UC-3500 and UF-4000 Sysmex UN Series Urinalysis; A Comparative Experimental Research Approach at an Urban Referral Hospital in Ghana

Abstract Background Urine analysis is one of the most common tests for assessing urinary-tract and kidney diseases. In recent years several automated instruments examining urine have been introduced. Though the introduction of automated urine analyzers is expected to reduce the labour involved, turnaround time and potential assay variations, manual urine analysis remains the gold standard or reference method for the analysis of urine. The new Sysmex UN-Series is believed to offer superior performance than other automated systems and manual methods. Methodology A total of 67 routine freshly voided midstream urine samples were analyzed within 1 hour in our study. The results of the automated system were compared with the results of the manual processing which consisted of physical, dipstick chemistry and sediment analysis. Correlation and concordance/agreement between the automated and the manual system were assessed by Bland-Altman plot and Cohen’s Kappa analysis respectively. Results There was a significant fair agreement between the manual analysis and Sysmex UF-3500, in the determination of urine physicochemical parameters (k = 0.193, P = 0.004). Both showed a significant moderate correlation (r = 0.593, P &amp;lt; 0.001) and a significant fair agreement (κ = 0.109, P &amp;lt; 0.001) for specific gravity and pH determination respectively. The best concordance between the two methods was in the nitrates (κ = 1.000, P &amp;lt; 0.001). Both systems recorded strong positive correlation for RBC (r = 0.951 and R2 = 0.904) and WBC (r = 0.91 and R2 = 0.822, P &amp;lt; 0.001) counts. Together, they had a significant good agreement [(65/67(97.0%), κ = 0.734, P &amp;lt; 0.001)] and a significant fair agreement [(48/67 (71.6%) κ = 0.065, P = 0.491)] for cast and bacteria counts correspondingly. Conclusion We conclude that the new automated Sysmex-UN series urine analyzer can be safely used in our laboratory since it demonstrated good performance and compatibility with the manual method. We recommend that microscopy results of the automated platform should be confirmed by manual microscopy, particularly in pathological samples.

Dengue Fever in India - A Review

Dengue fever is generally a mosquito borne viral infection which is transmitted from female Aedes Aegypti mosquito to human or infected human to mosquito, this virus is classified into five serotypes. Presenting features may range from asymptomatic to symptomatic (High grade fever, vomiting, hemorrhage, joint pain, headache ..etc.) Early and accurate diagnosis is critical to reduce mortality. Hematological tests includes RBC count, WBC count and Platelet count. We can protect ourselves from mosquito bites by using mosquito nets treated with Permethrin and getting rid of mosquito breeding sites. It is important to see a doctor when the symptoms become severe. Keywords: ,,, ,

A-380 Concordance of Truvian's Point-of-Care Blood Testing Analyzer to Central Laboratory Testing

Abstract Background Truvian has developed a fully automated blood testing analyzer (TruAnalyzer) capable of running a panel of clinical chemistry, immunoassay and hematology assays from a low-volume blood sample. Truvian's compact instrument was designed to deliver a convenient and affordable alternative to off-site traditional labs, providing lab-accurate rapid insights from a single, small sample of blood with no upfront pre-processing required to inform health decisions at the point of care. Here, we evaluate the precision and accuracy of a comprehensive wellness panel tested on the TruAnalyzer and compare its performance to FDA cleared devices. Methods TruAnalyzer performance was assessed for both precision and accuracy. For the precision studies, two commercially available controls (one for plasma-based assays and one for blood-based assays) were added to the Truvian consumables and loaded into the TruAnalyzer. Six replicates were run on each of the ten instruments (N = 60). Coefficient of variance (CV) analysis was performed and compared against pre-established requirements. For method comparison studies, 50 unique donors were consented and enrolled in an IRB approved study and 68 donor samples were collected over a 3-day period. For each donor, three 4-mL tubes of blood were drawn: 1) a Lithium Heparin (LiHep) sample for the TruAnalyzer clinical chemistry and hematology panel, 2) a LiHep gel tube for the Roche Cobas c311 clinical chemistry assays, and 3) an EDTA sample for the Sysmex PocH-100i complete blood count with 3-part differential. All samples were processed within 1 hour of collection. Using only 300 µL sample volume, the TruAnalyzer reported results for 23 analytes including CMP (no electrolyes), Lipid, HbA1c, and CBC with 3-part differential. Concordance to comparator devices was determined by Bland-Altman analysis. Results For the precision study, 13 out of 14 (93%) chemistry analytes had a between-system CV of ≤5.8%. For hematology, 5 out of 5 (100%) analytes had a between-system CV of ≤6%. For the method comparison study, RBC counts showed 98% of samples tested being within 6% of comparator values. WBC and platelet counts had 97% of samples tested being within 10% and 15% of comparator values respectively. 95% of hematocrit samples were within 6% of comparator values and 98% of hemoglobin samples were within 7% of comparator values. All hematology analytes showed equivalency to Sysmex PocH-100i as determined by Bland-Altman equivalency analysis. For clinical chemistry analytes, 14 out of 15 (93%) analytes showed equivalency to Roche Cobas c311 as determined by Bland-Altman equivalency analysis when CLIA ATE guidelines were applied. 7 out of 15 clinical chemistry analytes had ≥90% of results within ATE. 5 out of 15 clinical chemistry analytes had ≥80% of results within ATE. Conclusion The Truvian blood testing analyzer is the first multi-modality system capable of running hematology, clinical chemistry and immunoassays from a single, low-volume blood sample. Results from this study demonstrate good precision for the majority of panel analytes and excellent concordance to FDA cleared comparators for assays evaluated in this study.

A-145 Development of a Novel Algorithm Using Red Blood Cell Indices for Screening of Alpha Globin Gene Deletions

Abstract Background Distinguishing between different causes of microcytic anemia is essential for management of anemic patients. More than 20 different RBC indices have been used to screen thalassemia trait with inconsistent results. Differentiation between iron deficiency anemia (IDA) and alpha-thalassemia is challenging based on RBC indices alone. Moreover, thalassemia and IDA may coexist. A highly sensitive screening algorithm is needed to select clinical cases for alpha globin gene analysis to reduce unnecessary testing and improve pre-test probabilities. Methods A retrospective study was performed to evaluate the reliability of 22 RBC indices (RBC, MCV, RDW, MCH, Mentzer Index, RDW Index, etc) and formulae in differentiating cases with 1–2 alpha globin gene deletions from negative cases, in a cohort of 90 individuals with confirmatory alpha globin gene analysis results. Results The number of patients with each alpha globin gene deletion pattern were: 37 patients with alpha-thalassemia trait, 21 silent carriers (one gene deletion), 31 patients with negative result and 1 patient with triplicated alpha globin gene. The mean RBC count in each group were 4.79 ± 0.65 (trait), 4.45 ± 0.73 (carrier), and 4.21 ± 0.84 (negative) (million/mm). The mean MCV in each group were 71.1 ± 4.8 (trait), 76.1 ± 5.9 (carrier), and 74.2 ± 9.3 (negative) (fL). The mean RDW in each group were 16.5 ± 3.0 (trait), 17.2 ± 3.4 (carrier), and 18.3 ± 4.9 (negative) (%). The mean MCH in each group were 22.0 ± 1.8 (trait), 23.8 ± 2.3 (carrier), and 23.5 ± 3.7 (negative) (pg). None of the RBC indices showed significant difference between carrier and negative groups, while 20 RBC indices showed significant difference (P &amp;lt; 0.05) between trait and negative groups. The sensitivities of the RBC indices in detecting 1–2 alpha globin gene deletions based on published cut-offs ranged between 3.4% and 81.0%, while the specificities ranged between 41.9% and 100%. A novel scoring algorithm was evaluated based on the combination of 12 selected RBC indices and formulae, with each index or formula contributing to a score of 0 or 1. The average value of the negative cases was used as the score cut-off value for each index or formula. The scoring algorithm gave a score distribution between 0 and 12, with lower scores indicating alpha globin gene deletions. It achieved a sensitivity of 96.6% in detecting 1–2 alpha globin gene deletions, with a negative likelihood ratio of 0.15. The negative likelihood ratio of the scoring algorithm outperformed each of the individual RBC indices at similar levels of sensitivities (&amp;gt;95%). Conclusion The novel algorithm is a highly sensitive tool for screening of alpha globin gene deletions, including one gene deletion. It is more effective at identifying negative cases compared to individual RBC indices. Implementation of the algorithm could efficiently reduce 10.0% (9/90) of the test volume of alpha globin gene analysis. A more stringent version of the algorithm that incorporates standard deviation in the cut-off values could potentially achieve nearly 100% sensitivity and reduce 5.6% (5/90) of the alpha globin gene analysis test volume.

Hazardous Impact of Coal Dust on Hematological Parameters of Underground Coal Mine Workers

Due to the nature of their work, coal miners have historically faced significant exposure to large amounts of dust, placing them at a heightened risk. The present study was conducted to explore the hazardous effects of inhalation of coal dust on the haematological parameters of underground coal miners. 120 underground coal miners (60 workers having 5-10 years of experience and the other 60 having 10-20 years of experience) and 62 matched non-exposed to coal dust subjects as control from the locality were recruited for the study. After obtaining consent, different physiological parameters were measured, and blood was collected for assessment of haematological parameters. Observations revealed that there were lower mean values of total RBC count, haemoglobin level, MCV, PCV, MCHC and MCH in coal mine workers whereas mean values of Red Cell Distribution Width (RDW) and ESR were increased in the workers group. Further, coal dust exposure causes increased total leucocyte to count as well as a differential count of lymphocyte, neutrophil, monocyte, and eosinophil in the workers group but, a decreased differential count of basophils was detected among coal dust-exposed workers. All the changes in haematological parameters were found to occur experience-dependent and maximum changes were observed in workers having 10-20 years of working experience. There is a significant likelihood of coal mine workers experiencing changes in their blood composition, which suggests the harmful impact of coal dust on this group. Further, haematological parameters will help health professionals to screen any pathologic conditions and may help to prevent coal dust exposure-associated haematological disorders and complications thereof.