Differential WBC Counts Research Articles

Anti-allergic potentials of Ceriops decandra leaves in TDI-induced allergic mice: Comprehensive in-vivo and in-silico studies

BackgroundCeriops decandra is a mangrove plant, locally known as “Goran”. This plant has been traditionally used to treat hepatitis, liver disorders, ulcers, diarrhea, gastrointestinal disorders, hemorrhage, pain, inflammation etc. Here we investigated the safety and anti-allergic potential of C. decandra leaves. MethodThe toxicity was screened by mortality rate, and behavioral changes in mice. Anti-allergic activity test was conducted on TDI induced allergy model mice by assessing the common allergic symptoms such as sneezing, scratching, nasal score as well as total and differential WBC counts. HPLC-DAD analysis was used to determine the polyphenolic compounds in crude extract and performed in-silico molecular docking against four potential receptor molecules (Histamine H1, Histidine decarboxylate, Interleukin- 4 and 5). ResultsThe toxicity study of the extract on mice showed the innocuous nature of the plant even at the highest dose of 3000 mg/kg. In the anti-allergic assay, the leaf extract significantly (p<0.05) decreased the allergic symptoms along with the WBC counts in blood in a dose-dependent manner. The HPLC profile revealed the presence of six phenolic compounds (Catechin hydrate, Caffeic acid, Vanillic acid, Syringic acid, Rutin hydrate, p-coumarin). Among those Rutin hydrate, exhibited the best docking scores at all the targeted protein molecules and the highest binding affinity was (-9.8 kcal/mol) against Interleukin-5. ConclusionC. decandra is safe and rich in several polyphenolic compounds which are effective against allergic syndromes which could be an effective option for the management of allergic symptoms, although clinical studies are required to establish the herbal formulation.

Analytical Validation of Two Point-of-Care Assays for Hematological Analysis in the Miranda Donkey.

Miranda donkeys are an endangered autochthonous breed of Portugal. The HemoCue WBC DIFF and HemoCue Hb 201 portable analyzers, developed as a simplified alternative method for total and differential WBC count and hemoglobin measurement, respectively, may be relevant tools in veterinary practice. This study aimed to validate these instruments using Miranda donkey blood samples. For the HemoCue WBC DIFF, most parameters showed acceptable intra- and inter-assay precision with coefficients of variation (CV) below 5%, except for monocytes and eosinophils with higher CVs. The HemoCue Hb 201 showed CVs of 1.98% and 4.07%. Linearity correlation coefficients (r) ranged from 0.53 to 0.99 for HemoCue WBC DIFF and 0.99 for HemoCue Hb 201. Significant levels for neutrophils, lymphocytes, monocytes, eosinophils, and Hb measurements varied. Comparisons with ProCyte Dx showed an excellent correlation for WBC (rs = 0.96), neutrophils (rs = 0.91), lymphocytes (rs = 0.94), and eosinophils (rs = 0.90) but a poor correlation for monocytes and basophils. The HemoCue Hb 201 showed a correlation of rs = 0.96 with ProCyte Dx. In conclusion, both analyzers provided reliable results and are suitable for use in Miranda's donkey breed for WBC counts and Hb measurements.

Erroneous Automated WBC Differentials—A Case Series

Background and Aims: Peroxidase deficiency is one of the commonly inherited phagocytic defects. It has been also found to exist as a transient phenomenon in association with some clinical conditions. But that it can interfere and cause erroneous automated differential WBC count is something which is not commonly known. Materials and methods: Complete blood counts were analysed using Advia 2120i and Coulter DXH 900 haematology analysers and manually reviewed on peripheral blood smear stained with Leishman stain. Results: Peroxidase deficiency in neutrophils and eosinophils resulted in them getting counted as monocytes by the analyser causing pseudomonocytosis, pseudoneutropenia and pseudoeosinopenia. These were detected by a slide review and by reanalysing the samples on an analyser which worked on a different principle. Conclusion: There is a need to confirm monocytosis given by analysers working on the peroxidase principle with an alternate method. This will prevent needless medical investigations for pseudoneutropenia, pseudoeosinopenia and persistent monocytosis, thus preventing unwarranted mental agony and financial burden to patients. It also helps to save the laboratory’s reputation. A careful review of instrument flags not only helps reach an accurate result but sometimes they can also aid in the diagnosis of a rare potential genetic disorder like MPO deficiency.

Haematological and Demographic Features of Acute Myeloid Leukemia among Patients attending the National Oncology Center, Sana’a City - Yemen.

Background: The distinctive feature of acute myeloid leukemia (AML) is the infiltration of abnormal myeloid precursors into the bone marrow and other tissues, frequently accompanied by the appearance of these abnormal cells in the peripheral blood. This study aimed to describe the hematological features and demographic characteristics of patients with AML who attended the National Oncology Center in Sana’a, Yemen. Subjects and Methods: This descriptive study was carried out on 124 newly diagnosed AML patients of all ages and both genders who attended the National Oncology Center in Sana’a City, Yemen, during the period from November 2022 to November 2023. The complete blood count test was performed by drawing a few milliliters of blood sample, then was analyzed through a hematology analyzer, which performs quantitative analysis of blood elements including white blood cell count, WBC differential count, red blood cell count, hemoglobin, and platelet count. Results: The median age of the patients was 35 years and a range (0.9-779 years); the majority of cases were older than 18 years (62.9%); and the frequency of AML was higher in males than in females (57.26% and 42.74%), respectively. Of 124 newly diagnosed AML patients, the median (range) of WBC and platelets were 50.47 (1.18-534.32 103/uL) and 45 (343 103/uL), respectively. The mean SD for RBC was 2.85 0.78 106/uL,, Hb was 8.40.94 g/dL, and Blast count was 75.26 16.04%). Leukocytosis was detected in (62.9%), Hyperleucotyosis in (25%), Erythropenia in (87.1%), and Thrombocytopenia in (86.3%). There was a significant positive correlation between age and platelets (r = 0.194, p-value = 0.03), WBC and Blast cells (r = 0.285, p-value = 0.001), and highly significant between RBC and Hb (r = 0.623, p-value = &lt;0xC6&gt;&lt;0xB0&gt;0.001). For the WBC and platelet counts, there were variations between age groups and two genders; the variations were little in other hemological patterns, and there was no significance for all parameters. For WBC count, the median was 62 x 103 uL in the female group, higher than in the male group (46.5 x 103 uL), while there wasn't or little variation in the hematological features among male and female AML patients. Conclusion: Hematological features of bone marrow/peripheral blood are essential diagnostic tools in AML patients. In the present study, approximately 87% of patients present with leucotyosis or hyperleucotyosis, erythropenia, and thrombocytopenia. And the blast cell count was &gt; 20% for all AML patients, which is considered a diagnostic reference.

A comparison study between the Siemens ADVIA 120 and manual method for the differential white blood cell count in goats.

Although widely used, the ADVIA 120 hematology analyzer has not been previously validated for determining the differential leukocyte count in goats. The aim of this study was to compare the differential leukocyte counts provided by the ADVIA 120 (A-diff) and the manual method (M-Diff) in goats. EDTA blood samples that were analyzed within 4 h of collection were used in the study. The following exclusion criteria were applied: inappropriately filled tubes or tubes containing clots, erroneous ADVIA peroxidase cytograms, and blood smears of poor quality. The A-Diff was compared with the M-Diff performed by two independent observers on 200 leukocytes. Forty samples were included after previously excluding eight samples. The correlation between the A-Diff and M-Diff was very strong for eosinophils (r = .870, p < .001) and strong for lymphocytes (r = .796, p < .001) and neutrophils (r = .730, p < .001), while no significant correlation was observed for monocytes (r = .026, p = .872). The Passing-Bablok regression analyses revealed statistically significant constant errors for neutrophils (5.83%; 95% confidence interval [CI]: 0.41%, 12.18%) and eosinophils (1.89%; 95% CI: 1.17%, 2.71%). Bland-Altman analyses showed a statistically significant negative bias for lymphocytes (-5.0%) and a statistically significant positive bias for eosinophils (2.2%). The very low basophil percentages precluded a meaningful method comparison. The ADVIA 120 overall demonstrated good performance for the differential WBC count in goats under the conditions of this study. Therefore, it can be considered suitable for routine hematologic screening in goats. Nonetheless, it should be emphasized that any abnormal result should be confirmed with a blood smear evaluation.

Effects of Cyclosporine and Azacitidine on Some Hematologic and ‎Biochemical Parameters of Benzene-Induced Aplastic Anemia in Rats

Aplastic anemia, marked by deficiencies in hematopoietic stem cells, leads to peripheral blood pancytopenia and hypocellular bone ‎marrow. This study aimed to evaluate the therapeutic efficacy of cyclosporine and azacitidine, administered either alone or in combination, in rats with benzene-induced aplastic ‎anemia, focusing on restoring normal blood cell levels and preventing disease complications. Thirty adult female Wistar rats ‎(Rattus ‎norvegicus)‎ were randomly divided into five groups: negative control (C-, untreated), positive control (C+, induced aplastic anemia with ‎distilled water), cyclosporine-treated (CsA, 5.86 mg/kg), azacitidine-treated (Aza, 5.75 mg/kg), and combination-treated (CsA+Aza, 3.68 ‎mg/kg each). Benzene (1940 mg/kg) was administered orally for fifteen days to induce aplastic anemia. Post a 30-day treatment period, ‎evaluations included differential WBC and reticulocyte counts, serum IL-2 levels, and alkaline phosphatase (ALP) activity. Results ‎showed significant improvements in WBC% and reticulocyte% in all treated groups compared to the C+ group, with the combination-‎treated group showing the highest enhancement. IL-2 levels in the combination group were significantly reduced compared to other ‎treatment groups, aligning closely with the negative control. The ALP activity was significantly higher in both the cyclosporine and ‎azacitidine-treated groups compared to the positive control, with the combination group showing a marked increase over the azacitidine ‎group but no significant difference from the cyclosporine group and negative control.‎ In conclusion, the study demonstrates the potential therapeutic benefits of cyclosporine and azacitidine in treating benzene-induced ‎aplastic anemia in rats. The combination therapy, in particular, showed improved efficacy in all tested parameters, suggesting a potential ‎strategy for dose reduction and toxicity mitigation.

WITHDRAWN: Curcuma longa alone and in combination with pioglitazone attenuates paraquat-induced lung injury in rats through improving inflammation, oxidative stress and fibrosis

Background and aimThe therapeutic effects of pioglitazone (Pio), dexamethasone (Dexa) and Curcuma longa (Cl) on inhaled paraquat (PQ)-induced lung oxidative stress, inflammation, pathology changes and tracheal responsiveness were evaluated. Experimental procedureOne group of rats was exposed to saline (group C) and six groups to PQ aerosols. One group of PQ-exposed rats was given drinking water alone (group PQ), while other groups were received drinking water containing; two doses of Cl. (150 and 600 mg/kg, Cl-L and Cl–H), Pio (5 mg/kg), Pio + Cl-L and Daxa (0.03 mg/kg), (7 rat in each group). Results and conclusionIn PQ group, total and differential WBC counts, malondialdehyde (MDA) in the BALF, cytokine levels (INF-γ and IL-10), lung pathological changes and TR were remarkably increased but the BALF thiol, catalase (CAT) and superoxide dismutase (SOD) levels were reduced (for interstitial necrosis, p < 0.05 and for other cases, p < 0.001). Treatment of rats with Cl–H and Pio + Cl-L, significantly improved all measured variables, treatment with Pio and Cl-L improved most variables and treatment with Pio only few variables (p < 0.05 to p < 0.01). The improvements in almost all variables (except some pathological changes) in the Pio + Cl-L group were significantly higher than the Pio and Cl-L groups alone (p < 0.05 to p < 0.001).Curcuma longa ameliorated lung damage induced by inhaled PQ similar to dexamethasone and Pio and a synergic effect between Cl and Pio was observed, suggesting, amplified the activation effects of Pio on PPARs by Cl.

Modeling TCIRG1 Neutropenia By Utilizing Patient Derived Induced Pluripotent Stem Cells

Background: Severe congenital neutropenia (SCN) is a hereditary hematopoietic disorder characterized by recurrent infections and leukemic transformation. We previously identified a novel heterozygous missense mutation (R736S) in TCIRG1 causing SCN in a large multigenerational family (PMID: 24753205). We also identified a statistically significant correlation in individuals with lower ANC with rare missense variants in TCIRG1 in a large cohort of population-based genome screening (PMID: 27229898). Recently, we have discovered another TCIRG1 heterozygousvariant: R736C in two unrelated families with congenital neutropenia. Independently, Shinwari et al identified a novel TCIRG1 mutation, V52L, which was correlated with the neutropenic phenotype (PMID: 35573728). Homozygous mutations of TCIRG1 are causing autosomal recessive osteopetrosis (ARO), which is characterized by osteopetrosis and secondary hematological and neurological abnormalities. The Genome Aggregation Database (gnomAD) has no data about TCIRG1 R736S and R736C variants accruing in the healthy population. This is in contrast of numerous healthy individuals with a various heterozygous variants throughout the TCIRG1 gene. Aims: We aim to generate in-vitro model of TCIRG1 neutropenia by utilizing patient derived induced pluripotent stem cells (iPSCs) and characterize the hematological abnormalities caused by mutant TCIRG1 expression. We also aim to characterize the incidence of TCIRG1-associated neutropenia and the prevalence of TCIRG1 variants by sharing this new information within the scientific community through identifying more cases/families with this disease. Methods: Medical history, differential WBC counts and whole exome sequencing (WES) was used to identify the genetic basis of neutropenia in the family with R736S mutation. The R736C mutation in the new families also were identified by WES of the index cases. Medical history, blood counts and Sanger sequencing identified their affected relatives. iPSC lines of 4 affected individuals from 2 families harboring TCIRG1 R736S and R736C mutations and healthy volunteer were generated using standard methodology. S732F benign heterozygous TCIRG1 variant was generated by utilizing CRISPR/Cas9 knock-In gene editing using a healthy volunteer iPSC line. iPSCs hematopoietic differentiation was achieved by using STEMdiff Hematopoietic Kit. Resultant CD34+ cells were differentiated towards neutrophils using a published protocol (PMID: 35795780). Cell proliferation, survival and differentiation characteristics were measured by automated cell counter and flow cytometry, cell viability staining and surface markers analysis for granulocytic differentiation. Results: The discovery of these families came through the reputation and work of the Severe Congenital Neutropenia International Registry (SCNIR) and the expanding use of WES to identify the causes of hereditary disorders. With the identification of the index cases, we have identified several affected family members in each family. In all three families the severity of neutropenia is variable and correlates with the incidence of recurrent bacterial infections. Myeloid differentiation of CD34+ cells derived from iPSCs of 4 patients from two families with R736S and R736C mutations revealed maturation, survival and proliferation abnormalities compared to volunteer cells and cell line harboring benign S732F heterozygous TCIRG1 mutation. All patient cells had from 2 to 3-fold drop in cell proliferation and cell viability drop by over 70% for the patients with R736S and up to 90% with R736C mutation compared to the controls. The myeloid differentiation in the patients' cells, measured by flow cytometry and assessed by CD66b+/CD14+ and CD11b+/CD15+ markers was impaired by 38% and 23% respectively for the patients with R736S and 43% and 24% for the patients harboring R736C mutation. Conclusions: The pathogenesis of TCIRG1-associated neutropenia is currently unknown. Carriers of pathogenic variants from these families appear to have neutropenia with variable severity. Our preliminary studies show that patient iPSC derived hematopoietic cells closely recapitulate the neutropenic phenotype. Population based genomic studies; careful family histories and WES will identify new patients with this disorder.

Development and Pilot Testing of a Comprehensive Mobile Application to Assist Cell Count Determination During Peripheral Smear and Bone Marrow Examination.

In the modern era of complete blood count analysis, manual differential count is performed whenever 'flags' are generated by an automated hematology analyzer. Traditionally, tally counters with five or eight keys are used for manual differential count. A few mobile applications are available to perform this task; however, the application features and cell representation are limited. The primary objective of our study was to develop an indigenous, comprehensive mobile application to assist with manual blood cell differential count. The secondary objective was to measure the usability of a newly developed application among undergraduate medical students. A new mobile application was developed using a Java development kit, Version 11.0.13 (Oracle Corporation, Austin, USA) in Android Studio Dolphin (2021.3.1) (Google, California, USA). The application content was validated by three pathologists with more than five years of experience. The app's usability was tested among 60 participants using a validated mHealth App Usability Questionnaire (MAUQ). The questionnaire had 18 items covering three domains: ease of use, interface & satisfaction, and usefulness. The newly developed application supports peripheral smear WBC differential count, platelet count, reticulocyte count, malaria parasite quantification, and bone marrow differential count. During usability testing, the app was easy to use in 95% (57/60) of participants, time-efficient in 91.7% (55/60), and helpful for healthcare practice learning in 96.7% (58/60). The total mean score was 6.11, indicating high usability. A comprehensive mobile application to assist manual differential count with adequate cell representation was developed. The mobile application was easy to use, time-efficient, and valuable among the study participants.

Effect of resveratrol on haematological changes in diabetic-malaria infected wistar rats

The effect of resveratrol on haematological changes in diabetic-malaria infected rats was investigated. Five of six groups of male Wistar rats were induced with type 2 diabetes, followed by inoculation with malaria parasites. Four groups of the diabetic-malaria infected rats were orally given: low dose resveratrol (50 mg/kg), high dose resveratrol (100 mg/kg), Metformin (100 mg/kg) and Artesunate (5 mg/kg) respectively for six days after malaria inoculation. Administration of resveratrol, metformin and artesunate to diabetic-malaria infected rats significantly (p &lt; 0.05) decreased the blood glucose level (BGL) on days 3 and 6 post-inoculation. The high dose resveratrol (RSV 100) group showed a significant (p&lt;0.05) increase in red blood cell counts, although a significant (p &lt; 0.05) decrease of Packed Cell Volume was recorded in the low dose resveratrol group. There was no significant (p&gt;0.05) difference for the values of total WBC counts across the four groups of the diabetic-malaria infected rats, but the differential WBC counts (dWBCC) were significantly (p&lt;0.05) affected. High dose resveratrol administration significantly (p &lt; 0.05) reversed the dWBCC. The diabetic-malarial infected group showed significant (p&lt;0.05) decrease in average RBC; the average parasites and average infected RBC counts were significantly (p&lt;0.05) higher than that in the resveratrol groups. While, Metformin and Artesunate groups showed a further decline in parasitaemia. These findings indicate that Resveratrol decreased BGL and improved hematological parameters and suppressed parasitaemia in diabetic-malarial infected rats.

The effects of Crocus sativus extract on inhaled paraquat-induced lung inflammation, oxidative stress, pathological changes and tracheal responsiveness in rats

Paraquat is a green liquid toxin that is used in agriculture and can induce multi-organ including lung injury. Various pharmacological effects of Crocus sativus (C. sativus) were indicated in previous studies. In this research, the effects of C. sativus extract and pioglitazone on inhaled paraquat-induced lung inflammation, oxidative stress, pathological changes, and tracheal responsiveness were studied in rats. Eight groups of rats (n = 7 in each) including control (Ctrl), untreated paraquat aerosol exposed group (54 mg/m3, 8 times in alternate days), paraquat treated groups with dexamethasone (0.03 mg/kg/day, Dexa) as positive control, two doses of C. sativus extract (20 and 80 mg/kg/day, CS-20 and CS-80), pioglitazone (5 and 10 mg/kg/day, Pio-5 and Pio-10), and the combination of CS-20 + Pio-5 were studied. Total and differential WBC, levels of oxidant and antioxidant biomarkers in the BALF, lung tissue cytokine levels, tracheal responsiveness (TR), and pathological changes were measured. The levels of IFN-γ, IL-10, SOD, CAT, thiol, and EC50 were reduced, but MDA level, total and differential WBC count in the BALF and lung pathological changes were increased in the paraquat group (all, p < 0.001). The levels of IFN-γ, IL-10, SOD, CAT, thiol and EC50 were increased but BALF MDA level, lung pathological changes, total and differential WBC counts were reduced in all treated groups. The effects of C. sativus high dose and combination groups on measured parameters were equal or even higher than dexamethasone (p < 0.05 to p < 0.001). The effects of the combination of CS-20 + Pio-5 on most variables were significantly higher than CS-20 and Pio-5 alone (p < 0.05 to p < 0.001). C. sativus treatment improved inhaled paraquat-induced lung injury similar to dexamethasone and showed a synergistic effect with pioglitazone, suggesting possible PPAR-γ receptor-mediated effects of the plant.

A-134 Correlation Study of Haematology Analyser Sysmex XS 1000 vs Beckman DXH 520

Abstract Background An evaluation was conducted on 2 different haematology analysers to determine its result correlation. Both Sysmex XS1000i and Beckman DXH 520 are 5 part differential count analysers. Methods The evaluation was conducted on Sysmex XS 1000 and 2 units of Beckman DXH 520 at NHGD YIS lab in 2020. All instruments underwent precision study, carryover test and linearity to ascertain its performance prior to the concordance study.63 samples were used in the correlation all reportable parameters and WBC differential count and its flagging. Results The 2 units of DXH 520 demonstrated acceptable performance in precision, carryover and Linearity. The correlation of the parameters were within RCPA allowable error. However, we noted a low concordance of 39% in the WBC differential count flagging. To validate the flagging, manual differential count was conducted. Conclusion During the correlation, Sysmex XS appeared to be more sensitive to Atypical lymphocytes where manual differential count read &amp;lt;5%. This inevitably resulted in more manual slide review. In the 63 samples, 4 cases with high atypical lymphocytes (&amp;gt;5%) seen in manual differential count and they were flagged as lymphocytes in sysmex XS1000i and as monocytes in DxH 520. To counter the potential missed out in the reporting of Atypical lymphocytes in DXH, a lower threshold for monocytes in slide review may be needed.

Evaluation of canine and feline leukocyte differential counts obtained with the scil vCell 5 compared to the Advia 2120 hematology analyzer and a manual method.

The vCell 5 (scil Animal Care), a point-of-care hematology analyzer (POCA), was recently introduced to veterinary laboratories. This laser- and impedance-based analyzer is capable of providing a CBC with 5-part WBC differential count (Diff) along with WBC cytograms and flags serving as interpretation aids for numerical results. We compared the scil POCA-Diff to reference methods (i.e., manual differential count, Advia 2120 hematology analyzer [Siemens]) for canine and feline blood samples and considered WBC cytograms and flags. Total observed error (TEo), calculated from CV and bias%, was compared to total allowable error (TEa). Data were analyzed before and after a review process (exclusion of flagged and samples with invalid cytograms). For both species, correlation was good-to-excellent (rs = 0.81-0.97) between both analyzers for all variables, except for feline monocytes (rs = 0.21-0.63) and canine monocyte% (rs = 0.50). Smallest biases were seen for neutrophils (dog: -5.7 to 0.8%; cat: 1.5-9.4%) with both reference methods. Quality requirements (TEo < TEa) were fulfilled for canine and feline neutrophils (TEo = 5.3-10.6%, TEa = 15%) and eosinophils (TEo = 67.1-83%, TEa = (90)-50%) considering at least one reference method. Our review process led to mildly higher rs-values for most variables. Although not completely satisfactory, the scil POCA provides reliable results in compliance with ASVCP quality goals for canine and feline neutrophils and eosinophils. Analyzer flag and cytogram analysis served as useful tools for QA, indicating the necessity for manual review of blood smears, and contributed to improvement of scil POCA performance.

Evaluation of nano-curcumin against inhaled paraquat-induced lung injury in rats.

Acute lung injury (ALI) remains a significant source of morbidity and mortality in critically ill patients and currently there is no efficient therapy for this condition. The aim of this research was to evaluate the protective activity of nano-curcumin (nano-CU) as a natural anti-inflammatory and antioxidant agent, against inhaled paraquat (PQ)-induced lung injury. One group of rats was exposed to saline (control group, Ctrl) and six groups to PQ aerosol (54mg/m3 on alternate days 8 times, each time for 30min) treated with drinking water alone (group PQ), 2 and 8mg/kg nano-CU (nano + CU(L) and nano + CU(H)), 5mg/kg pioglitazone (PIO), nano-CU(L) + PIO or 0.03mg/kg dexamethasone (Dexa) for 16days after PQ exposure period. PIO and Dexa were intraperitoneal (ip) injected and nano-CU was administered orally (po), (6 rats in each group). In the PQ group, total and differential WBC counts, malondialdehyde (MDA) in the bronchoalveolar lavage fluid (BALF), interferon gamma (INF-γ) and interleukin 10 (IL-10) levels in the lung tissues, lung pathological changes, and tracheal responsiveness were increased but the BALF thiol, catalase (CAT) and superoxide dismutase (SOD) levels were reduced. In treated groups with nano-CU(H) and PIO + nano-CU(L), all measured variables, in Dexa and nano-CU(L) treated groups, most variables and in the PIO group only a few variables were improved. The improvement of most variables in the PIO + nano-CU(L) group was significantly higher than in the PIO and nano-CU(L) groups alone. Nano-CU ameliorated lung damage induced by inhaled PQ similar to dexa and a synergic effect between nano-CU and PIO was observed, suggesting, a possible PPAR-γ receptor-mediated effect of curcumin.

Paraquat-induced systemic inflammation and oxidative stress in rats improved by Curcuma longa ethanolic extract, curcumin and a PPAR agonist

The effect of Curcuma longa (Cl) ethanolic extract, nano-curcumin (Cu) and a PPARγ activator, pioglitazone on inhaled paraquat (PQ)-induced systemic inflammation and oxidative stress was examined in the present study. Control rats were exposed to normal saline and PQ groups to 27 and 54 mg/m3 (PQ-L and PQ-H) aerosols. Nine other PQ-H groups were treated with Curcuma longa (Cl, 150 and 600 mg/kg/day), nano-curcumin (Cu, 2 and 8 mg/kg/day), pioglitazone (Pio, 5 and 10 mg/kg), low dose of Pio + Cl and Cu and dexamethasone (0.03 mg/kg/day) for 16 days after PQ exposure period (n = 8). Total and differential WBC counts, malondialdehyde (MDA) and TNF-α levels were increased but thiol, catalase (CAT), superoxide dismutase (SOD), IL-10 and IFN-γ levels were decreased in the blood in the both PQ groups (p < 0.05 to p < 0.001). Treatment with Dexa and both doses of Cl, Cu, and Pio improved all measured variables compared to the PQ-H group (p < 0.05 to p < 0.001). The improvements of most variables in the treated group with low dose of Pio + Cl and Cu were higher than the effects of three agents alone. Systemic inflammation and oxidative stress induced by inhaled PQ were improved by Cl, Cu and Pio. In addition, a synergic effect between Pio with those of Cl and Cu was shown, suggesting PPARγ mediated effects of the plant and its derivative Cu.

Novel TCIRG1 Mutations Causing Congenital Neutropenia

Background: Severe congenital neutropenia (SCN) is a bone marrow failure disorder leading to severe neutropenia related infections and leukemia. More than 30 different genes associated with this disorder have been identified. We previously identified a novel heterozygous missense mutation (R736S) of TCIRG1 as the probable cause for SCN in a large multigenerational family through exome sequencing (Makaryan et al; Hum Mutat. 2014 Jul;35(7):824-7). We also identified a statistically significant correlation of rare missense variants of TCIRG1 with lower ANC in three cohorts from the NHLBI Grand Opportunity Exome Sequencing Project (GO ESP; Rosenthal et al; Genet Epidemiol. 2016 Sep;40(6):470-4). Recently, we have discovered a novel TCIRG1 variant (R736C) at the same position in a new family with congenital neutropenia. TCIRG1 encodes subunit a3, a 116-kD component (OC116) of V-ATPases, vacuolar proton pumps which mediate H+ transport through membranes. Homozygous loss-of-function mutations of TCIRG1 are the most common cause of autosomal recessive osteopetrosis (ARO). Mutations R736S and R736C occur at an amino acid residue critical for proton translocation. There are no reports in the literature of individuals with heterozygous mutations at this locus. This is in contrast to the numerous healthy individuals with various heterozygous variants throughout the TCIRG1 gene. Parents of ARO patients harboring homozygous TCIRG1 mutations are considered healthy with no musculoskeletal or hematopoietic abnormalities. Pathogenesis of TCIRG1-associated neutropenia is largely unknown. Hypothesis: We hypothesize that certain heterozygous TCIRG1 variants, like R736S and R736C, may cause SCN through specific molecular pathways. Methods: Family and medical history, differential WBC counts and whole exome sequencing/Identity by Decent (IBD) analysis were used to identify the genetic basis of neutropenia in the previously reported family. Sanger sequencing was used to confirm the findings of exome sequencing and to identify other affected individuals. The R736C mutation was identified by whole exome sequencing with further site-specific analysis of 35 genes. Affected relatives were identified by Sanger sequencing. The Polyphen 2 online prediction tool was used to predict the impact of both substitutions on TCIRG1 function. Results: Discovery and details of the original family were described in our previous report: (Makaryan et al; Hum Mutat. 2014 Jul;35(7):824-7). The second family was discovered in Taiwan, where the primary physician contacted SCNIR about a young patient with neutropenia who had a novel TCIRG1 mutation. Shortly thereafter, we identified two more affected family members. In both families the severity of recurrent bacterial infections is variable, roughly correlating with the severity of the neutropenia. For example, the grandmother of the index patient in the second family was not aware that she had neutropenia. The novel R736C TCIRG1 variant is located in the same position as the previous R736S variant. Arginine in this position is evolutionary highly conserved and both these substitutions cause a major change in amino acid size and properties. Polyphen-2 analysis predicts the effects of both mutations "probably damaging” (score 1.0). Conclusions: Understanding the genetic, epidemiological and molecular basis of TCIRG1-associated neutropenia is important for improving the diagnosis and treatment of this form of congenital neutropenia. The pathogenesis of TCIRG1-associated neutropenia is unknown. It is very important to emphasize that carriers of pathogenic variants from both families have neutropenia with variable severity. The report of this new family suggests that there may be other patients and families with neutropenia caused by mutations in TCIRG1. Population based genome studies, careful family histories and whole genome/exome sequencing will be highly valuable for identifying and helping new patients with this disorder.

Effect of microbial phytase and Nigella sativa seed extract supplementation on some hematological parameter in albino rats treated to cadmium chloride toxicity

The present study was conducted to determine the role of microbial phytase and Nigella sativa seed alcoholic extract in relieving the toxicological effects of cadmium chloride in some hematological parameters (RBC count, Hb concentration, PCV, total WBC count, Differential WBC count, and concentration of ALT, AST, &amp; ALP). Fourty mature male rats were randomly divided into 4 equal groups (control and three treated groups). All rats were drenched CdCl222 solution (30 mg/L).T1 supplemented with the microbial phytase (500 mg/kg of provender), T2 drenched the black seed extract (200 mg/L). and T3 received both of treatments. Eight weeks later, blood samples were obtained by heart puncture for estimation of the hematological parameters included in the present study. The results revealed a significant increase (P&lt;0.05) in RBC count, Hb concentration and PCV of T1, T2 and T3 groups compared with control especially in T3, while there is no significant differences (P&gt;0.05) in total WBC count. Neutrophils &amp; eosinophils numbers showed a significant decrease in T1, T2 &amp; T3 groups when compared with control, while lymphocytes showed an opposite results. ALT, AST, &amp; ALP concentrations in T1, T2 &amp; T3 groups showed a significant decrease (P&lt;0.05) in comparison with control, especially that of T3. It can be concluded that the microbial phytase supplementation and Nigella sativa seed alcoholic extract drenching have an efficient role in relieving the toxicological effects of CdCl222 in rats especially when supplemented with each other.

Neopterin as an Inflammatory Biomarker in Patients with Non-alcoholic Fatty Liver Disease

Background: Early diagnosis of non-alcoholic fatty liver diseases (NAFLD) through detection of a new biomarker is crucial to prevent disease deterioration. Aim: To evaluate the serum level of neopterin in NAFLD patients and to demonstrate its role as an early biomarker of NAFLD.Methodology: This is an observational, cross-sectional study that conducted on 71 newly diagnosed NAFLD patients and 60 healthy individuals as a control group. Liver enzymes (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]) and uric acid were determined. The serum level of neopterin was measured by ELISA and WBC differential count by a Coulter counter analyzer. Erythrocyte sedimentation rate (ESR) was determined by Wintrobe’s method. Results: NAFLD patients had a significantly higher BMI, waist circumference, and body weight compared to control, also their blood pressure was significantly higher. The serum level of ALT, AST, and uric acid were significantly higher in NAFLD patients than the control. There is non-significant difference between NAFLD patients and controls regarding ESR, lymphocytes and granulocytes, while monocytes and neopterin values were significantly higher among NAFLD patients.Conclusions: Serum neopterin level can be a predictor inflammatory marker in NAFLD patients. Clinical Pharmacists as part of healthcare professionals can support NAFLD patients by educating them on lifestyle modification if the disease is diagnosed early before reaching a critical stage.

The expression of circulating hsa-miR-126-3p in dengue-infected Thai pediatric patients

ABSTRACT Circulating hsa-miRNA-126 (CmiR-126) has been reported to involve in the pathogenesis of many infectious diseases including dengue virus infection. However, no prior study has been conducted to describe more details in dengue-infected pediatric patients. This study aimed to describe CmiR-126-3p in dengue-infected pediatric patients during the febrile and convalescent phases. Additionally, the correlations between CmiR-126-3p and other relevant clinical laboratory factors were investigated. Sixty paired-serum specimens collected during febrile and convalescent phases were retrieved from patients with dengue fever (DF) (n = 30) and dengue hemorrhagic fever (DHF) (n = 30). Thirty paired-serum specimens collected from non-dengue acute febrile illness patients (AFI) were included as the control group. CmiR-126-3p was determined using reverse transcription quantitative real-time polymerase-chain reaction (RT-qPCR). Relative miRNA expression was calculated as 2−ΔCt using CmiR-16-5p for data normalization. CmiR-126-3p expression during febrile and convalescent phases in dengue-infected patients was significantly lower than AFI (p < 0.05). However, miRNA levels were not different (p > 0.05) compared between DF and DHF and between primary and secondary infection. CmiR-126-3p levels in DF in the convalescent were significantly higher than in the febrile phase (p = 0.025). No association between CmiR-126-3p and hematocrit, WBC level, platelet count, WBC differential count or dengue viral load was observed (p > 0.05). The data suggest that hsa-miR-126-3p involved in pathogenesis of dengue infection and may be a promising early and late biomarker for DENV infection. However, hsa-miR-126-3p alone cannot be used as a predictor for dengue severity.

Environmental, Biochemical and Hematological study in rats exposed to Cadmium Chloride in Drinking Water and the role of Vitamin E and C

The present study was carried out to determine the synergistic role of vitamin E and vitamin C in reducing the toxic effects of cadmium chloride on some blood parameters (RBC count, Hb concentration, PCV, Total WBC count, Differential WBC count), as well as some biochemical parameters which include (Malondialdehyde (MDA), Aspartate aminotransferase(AST), Alanine aminotransferase (ALT), Alkaline phosphatase(ALP), Creatinin, Urea) and some antioxidants (Superoxidedismutase(SOD), Glutathion-S-transferase(GST), Catalase(CAT). Forty adult male rats were divided into five equal groups. The first group was left without any treatment (the control group), the second was given cadmium chloride (30mg/L), the third was given cadmium chloride and vitamin E simultaneously, the fourth was given cadmium chloride and vitamin C simultaneously, the fifth was given cadmium chloride, vitamin E and vitamin C simultaneously. The results showed a significant decrease (P <0.05) in (RBC count, Hb concentration, PCV) and a significant increase (P <0.05) in WBC count (neutrophils and eosinophils). On the other hand, the results also showed an increase (P <0.05) in the concentration of (ALT, AST, ALP, Urea, Creatinine, and MDA)and a significant decrease in antioxidants(SOD, CAT, GST) in (T1) compared with the control group. On the other hand, the results showed a noticeable improvement in the parameters studied when using vitamin E and vitamin C alone or together with cadmium chloride, and this indicates the therapeutic role played by vitamins in reducing the toxicity caused by cadmium chloride