Normal Erythrocytes Research Articles

2644 ACE Inhibitor-Induced Visceral Angioedema - A Rare Phenomenon

INTRODUCTION: Angiotensin Converting enzyme (ACE) inhibitors are widely used to treat hypertension and congestive heart failure. Common adverse reactions include cough, hypotension and dizziness. Angioedema is a rare reaction that occurs in less than 1% of patients. Most cases of angioedema are involving the lips, tongue, face and airway. More rarely, visceral angioedema can also occur with jejunal involvement. CASE DESCRIPTION/METHODS: A 24-year-old female with recently diagnosed hypertension treated with lisinopril presented for evaluation of abdominal pain, vomiting, and diarrhea. On examination, she had moderate epigastric tenderness with normal bowel sounds and no organomegaly. Labs revealed white blood cells 20 k/mm3, elevated C-reactive protein 34.2 mg/L and normal erythrocyte sedimentation rate 7 mm/hr. Complement levels including C1 esterase inhibitor, C1q and C4 were found to be normal. Stool culture also remained negative. Computed tomography (CT) of the abdomen revealed moderate ascites, marked mucosal thickening of jejunal and ileal loops with adjacent mesenteric edema (Figure 1). Magnetic Resonance Enterography did not reveal any pathological process. A paracentesis was performed with the ascetic fluid analysis revealing a low serum-ascites albumin gradient without infection. Endoscopy with small bowel enteroscopy revealed normal duodenum and jejunum (Figure 2). Jejunal biopsy returned negative for an infectious or inflammatory process. Due to negative diagnostic workup, a diagnosis of ACE inhibitor-induced visceral angioedema was suspected. Patients symptoms quickly improved 24 hours after cessation of lisinopril and she was discharged home. On follow up 4 weeks later, the patient remained symptom-free. DISCUSSION: Visceral angioedema is a rare side effect of ACE inhibitors most commonly affecting the small bowel. The jejunum is most often affected followed by ileum and duodenum. High levels of bradykinin can cause increased vascular permeability within the bowel mucous membrane causing edema. Symptoms commonly include abdominal pain, anorexia, vomiting and diarrhea. Symptoms typically begin 72 hours after starting the ACE inhibitor. Imaging studies show circumferential small bowel thickening with mesenteric edema and ascites. Endoscopic examination is usually normal. Diagnosis is made by exclusion of other etiologies and improvement of symptoms within 48 hours of drug cessation. Treatment includes discontinuation of the offending drug and supportive therapy until symptoms are resolved.

Effect of Cerium on Erythrocyte Mechanical Properties

Lanthanide’s are known as rare earth elements and are used in many areas. Although the place of Cerium in the Lanthanide family in its biological importance in mammalian physiology is not fully known, a human weighing 70 kg contains about 40 mg Cerium. Soluble Ce3+ salts (nitrate, acetate, chloride, etc.) are used for many biomedical purposes and in particular act as calcium analogues in biological systems. In non-physiological flow conditions occurring in extracorporeal circulation areas, increased calcium ions within the cell play a role in reducing erythrocyte deformability. In this study, it is aimed to show the possible effects of Cerium on changes in erythrocyte deformability in shear stress at sub-hemolytic level. For this purpose, blood samples were taken from adult male volunteers. Normal erythrocyte deformability was measured at nine shear rates between 0.5 and 50 Pa, followed by stress application at a slip rate of 290 Pa for 300 seconds without removing the sample from the system, followed by erythrocyte deformability at shear rates of 0.5-50 Pa. There was no significant difference between the erythrocyte deformability values of the control and Cerium incubated blood samples before stress application. However, significant differences were found after stress application (p <0.01). Incubation of erythrocytes with Cerium did not cause any change in erythrocyte deformability, but caused further deterioration of erythrocytes after mechanical stress. Cerium chloride, which we use in our study, should be kept in mind that under normal conditions, even if there is no negative effect on erythrocyte mechanics, conditions change, erythrocytes are exposed to short-term sub-hemolytic mechanical effects and their mechanical properties are deteriorated. Keywords: Lanthanide, Cerium, Erythrocyte, Deformability, Mechanical Stress DOI: 10.7176/JSTR/5-8-11

Obez çocuklarda alkolik olmayan yağlı karaciğer hastalığı ve eşlik eden diğer karaciğer hastalıkları

Objective: The prevalence of non-alcoholic fatty liver disease (NAFLD) in children increased parallel to the increment of childhood obesity. Also, NAFLD may be the presenting feature of different liver diseases in non-obese children. We aimed to determine whether there were co-existing liver diseases in overweight and obese children with NAFLD. Material and Methods: Pediatric gastroenterology outpatient clinic records of obese and overweight patients, aged between 5-18 years, were retrospectively reviewed. Seventy patients who had liver steatosis on ultrasonography, and alanine aminotransferase (ALT) levels were higher than 1.5 times the upper limit of normal were recruited to the study. The demographic findings, laboratory tests for infectious, metabolic, and autoimmune causes, abdominal ultrasonography and liver biopsy findings of patients were recorded. Results: At presentation, 94.2% of the patients (n: 66) had mild transaminase elevation. All patients were negative for viral hepatitis, anti-tissue transglutaminase immunoglobulin (Ig) A, anti-liver-kidney-microsome type 1 and anti-smooth muscle antibody. They had normal erythrocyte sedimentation rate (ESR) , C-reactive protein (CRP), ceruloplasmin, and total IgG levels . O nly one patient with low alpha-1 antitrypsin levels had heterozygotes of the PiMZ phenotype. Three (4.3%) patients had antinuclear antibody ( ANA) positivity. 44.7% of patients were given ursodeoxycholic acid treatment. On follow-up, normalization of ALT was achieved in 31 (44.2%) patients at mean 6.1 ±4.6 (2-19) months, but no relation was found between normalization and ursodeoxycholic acid treatment. A patient with ANA positivity had increased ALT, ESR , CRP , IgG levels and ANA titers o n follow-up, and she was diagnosed with autoimmune hepatitis with the support of liver biopsy. Under prednisolone and azathioprine treatment, ESR and IgG levels were normalized, ALT and ANA titers decreased. Conclusion: Other causes of chronic hepatitis should be screened in obesity-related non-alcoholic fatty liver disease and the development of autoimmune hepatitis should be kept in mind in the presence of ANA.

PS1525 ANTISICKLING EFFECTS OF 1,2,3‐TRIAZOLE DERIVATIVES AS POTENTIAL NEW DRUGS FOR SICKLE CELL DISEASE

Background:Sickle cell disease (SCD) is a monogenic hereditary hematologic disease, being the most common in Brazil, in which about 3.500 children are born each year with it (Felix et al, 2010). It is characterized by the substitution of adenine for thymine, encoding valine rather than glutamic acid, at position 6 of the β‐globin chain. In deoxygenation, the erythrocytes containing the mutated hemoglobin, hemoglobin S (HbS), take on the sickle shape due to Hbs polymerization (Eaton &amp; Hofrichter, 1990). In Brazil, only the use of hydroxyurea (HU) is allowed by Agência Nacional de Vigilância Sanitária (ANVISA). However, in 2017, the commercialization of L‐glutamine was permitted by the Food and Drug Administration (FDA) in the United States of America, which proved to be effective in reducing the main acute effects of SCD, preventing the oxidative stress (Gardner, 2018). Recent studies have revealed a previously unrecognized role of adenosine in normal and sickle erythrocytes, the adenosine signaling in the regulation of 2,3‐bisphosphoglycerate (2,3‐BPG) production, an erythrocyte‐specific metabolite that induces the release of O2. And in 2012, Carlsson et al. verified the activity of new 1,2,4‐triazoles that showed, among activities in other adenosine receptors at varying levels, ADORA2B receptor antagonistic activity.Aims:The main objective of this work is to study the effect of the 1,2,3‐triazolic substances NHSulf, NHSulfNH2, NHSulfCl and NHSULfCH3 on erythroid physiology and pathophysiology in normal and sickle cell patients.Methods:Four experiments were performed with samples from control patients and sickle cell patients. First the samples are centrifuged for separation of the erythrocytes and the buffy coat removed, washed with PBS, this process is repeated three times and is done in all experiments. In the osmotic fragility assay the samples are placed with control, DMSO, or HU, or the substances in increasing concentrations of saline (NaCl 0.1% to 0.9%), and then read at 540 nm. 2.3‐BPG levels are measured with the Roche comercial Kit. The erythrocyte sickling reversion test with sickling induced by the addition of sodium metabisulfite (MB) (for 2 h) and the substances are added (for 2 h), and the reversal of the sickle cells was verified in 400x magnification, and the percentage of sickle cells compared to control. Polymerization inhibition was also verified, adding to the samples, DMSO, or HU, or the substances and then adding the MB, and reading in Elisa at 700 nm.Results:Osmotic fragility test and 2,3‐BPG levels were done with control patients and was observed that substances NHSulf, NHSulfNH2, presented statistically lower osmotic fragility in osmotic stress (lower saline concentrations). NHSulfCl and NHSULfCH3 did not present significant difference compared to control. As for 2,3‐BPG levels the samples presented a dose‐dependent result to different drugs concentrations. Reversal sickling test and inhibition of polymerization showed that substances (0.33 mM) have a potential of sickling reversion when compared to control and HU. For polymerization inhibition it can be seen that the molecules decreased polymerization significantly when compared to control, and possess greater potential when compared to HU.Summary/Conclusion:Thus, we observed that the substances NHSulf, NHSulfNH2, NHSulfCl and NHSULfCH3 has the potential to decrease osmotic fragility and to increase 2,3‐BPG levels in control erythrocytes, and to reverse the sickling and to inhibit polymerization in sickle cell patients.image

Manilkara zapota (L.) P. Royen Leaf Extract Derived Silver Nanoparticles Induce Apoptosis in Human Colorectal Carcinoma Cells Without Affecting Human Lymphocytes or Erythrocytes.

Plant-derived synthesis of silver nanoparticles (AgNPs) has found wide biomedical applications including cancer cure. This report deals with biosynthesis of silver nanoparticles (MZLAgNPs) employing leaf extracts of Manilkara zapota (L.) under optimized conditions. Characterization of MZLAgNPs using UV-Vis spectroscopy, FTIR, XRD, and FESEM analyses revealed that the particles were predominantly spherical averaging 24nm in size. Their cellular effects were assessed by MTT assay, fluorescence, and scanning electron microscopy of cells stained with propidium iodide, acridine orange/ethidium bromide, and annexin V-FITC to visualize signs of apoptosis. Evaluation of cell proliferation by clonogenic assay, wound healing ability by scratch assay and cell cycle distribution by flow-cytometry was also carried out. Apoptosis-related gene expressions were analyzed by RTq-PCR and western blot analysis. MZLAgNPs selectively inhibited growth of colorectal carcinoma HCT116, HeLa, and non-small lung carcinoma A549 cells, dose-dependently with IC50 concentrations of 8, 16, and 29μg/mL respectively, following 72-h treatment, without affecting growth of normal human lymphocytes and erythrocytes. Apoptosis induction was observed by fluorescence and scanning electron microscopy. Overproduction of reactive oxygen species (ROS), reduction of mitochondrial membrane potential, upregulation of apoptotic-related genes - PUMA, cas-3, cas-8, cas-9, and BAX, expression of caspase 3, and occurrence of PARP cleavage were observed in MZLAgNPs/cisplatin treated cells. Taken together, our results clearly demonstrate the therapeutic potential of biogenic MZLAgNPs as an effective agent for killing colorectal carcinoma cells by apoptosis induction.

Study on angular distribution of differential photoacoustic cross-section and its implication in source size determination.

Angular distribution of a differential photoacoustic cross-section (DPACS) has been examined for various nonspherical axisymmetric particles. The DPACS as a function of measurement angle has been computed for spheroidal particles with varying aspect ratios and fitted with a tri-axes ellipsoid form factor model to extract shape parameters. Similar study has been carried out for normal and pathological red blood cells, and fitting has been performed with the tri-axes ellipsoid and finite cylinder form factor models to evaluate cellular morphology. It is found that an enhancement of the DPACS occurs as the surface area of the photoacoustic source normal to the direction of measurement is increased. It decreases as the thickness of the source along the same direction increases. For example, the DPACS for normal erythrocyte along the direction of symmetry is nearly 20 times greater than a pathological cell. Further, the first minimum appears slightly later (≈4°) for a healthy cell compared with that of a diseased cell. Shape information of spheroids can be precisely estimated by the first model. Both models provide accurate estimates of shape parameters for normal red blood cells (errors within 4%). It may be possible to assess cellular morphology from an angular profile of the DPACS using form factor models.

No Relation Between Zinc Status and Inflammatory Biomarkers in Adolescent Judokas.

This study aimed to assess the relation between zinc status and inflammation biomarkers in adolescent judokas. This cross-sectional study included 52 male adolescents, aged between 14 and 19 years, who were subdivided into two groups: judoka group (n = 25) and control group (n = 27). Zinc intake was monitored using 3-day food records and the NutWin software version 1.5. The plasma and erythrocyte zinc concentrations were determined by flame atomic absorption spectrophotometry. Analysis of cytokines (IL-1β, IL-6, and TNF-α) was performed. The mean values of zinc concentration in the diet were found to be higher than those recommended (11.0±3.9 mg/day and 20.3±11.9 mg/day for control group and judokas, respectively) although there was no significant difference between the groups. The mean plasma concentrations of zinc were below the reference range (71.4±16.0 μg/dL and 71.9±13.8 μg/dL for control group and judokas, respectively), without a significant difference between the groups. The mean concentrations of zinc erythrocyte were within the reference range (41.2±8.6 μg/gHb and 42.6±11.3 μg/gHb for control group and judokas, respectively), without a significant difference between the groups. There was no significant difference in the inflammatory biomarkers between the judokas and controls. There was not a significant correlation between biochemical parameters of zinc and inflammation biomarkers in adolescent judokas. Regarding the data found in the study, it can be concluded that the athletes evaluated have low plasma zinc concentrations, normal erythrocyte values, and high dietary intake of zinc. Moreover, the study don't show a relationship between zinc parameters and inflammatory markers evaluated.

Evaluation of Cytotoxic Activity of Annona muricata Fruits and Leaves

Introduction: Screening of various medicinal plants for biological active compounds has become a vital source of cancer related drugs. The present study was designed to evaluate and compare the cytotoxicity of Annona muricata fruits and leaves. Method: The leaves and fruits are dried and standardised by performing ash value, extractive value and phytochemical screening, the dried fruits and leaves of Annona muricata were defatted with petroleum ether and extracted using methanol as solvent. The extracts were evaluated for in-vitro cytotoxic study by MTT assay using Hep G2 cell lines and trypan blue dye exclusion method using DLA and EAC cells. Haemolytic activity was performed to determine the toxicity of extracts to normal cells. Results: The qualitative analysis of phytochemical extracts and parts possesses carbohydrates, cardiac glycosides, alkaloids, flavones and tannins. In MTT assay, while comparing the cytotoxic action of methanolic extracts of Annona muricata leaves and fruits of (MEAML and MEAMF) with standard curcumin (18.6µg/ml), MEAMF (IC50=12.4 µg/ml) showed more cytotoxic action to Hep G2 cell lines than MEAML (IC50=82.5µg/ml). In dye exclusion test, MEAML showed an IC50 value of about 140.3µg/ml, 137.2µg/ml for DLA and EAC respectively, whereas 179.4µg/ml, 165.2µg/ml in MEAMF. None of the extract possessed any haemolytic activity revealing their non-toxicity on normal cells. Conclusion: Methanolic extracts of Annona muricata fruits and leaves possess cytotoxic activity with no significant haemolytic activity towards normal erythrocytes.

Hands as cold as ice

A healthy 24-year-old woman presented with a 1-year history of discolouration, paraesthesia and pain predominantly affecting the middle and ring fingers of both hands. Symptoms were attributed to cold exposure...

RELATIONSHIP BETWEEN PESTICIDE EXPOSURE AND HEMOGLOBIN LEVEL AND ERYTHROCYTE AMOUNT IN HORTICULTURAL FARMERS IN THE DISTRICT OF PAAL MERAH, JAMBI CITY

Background: The District of Paal Merah, Jambi City, is a center for vegetable production for the Jambi City area and its surroundings. The aspect of excessive use of pesticides by farmers in the District of Paal Merah, Jambi City is a problem that needs attention, especially the practice of spraying which ignores aspects of safety and health of farmers that can lead to health problems both directly and indirectly for farmers. One of the long-term effects caused by exposure to pesticides is a decrease in hemoglobin levels and the amount of erythrocytes in the blood. Based on the report of the Payo Selincah Health Center in Jambi City, it was explained that in the District of Paal Merah, Jambi City in 2016 showed anemia sufferers reached 30% of the number of public health center visits.&#x0D; Objective: The purpose of this study was to describe and analyze the relationship of exposure to pesticides with hemoglobin levels and the number of erythrocytes in horticultural farmers in the District of Paal Merah, Jambi City.&#x0D; Method: This type of research is observational analytic with cross sectional study design conducted in the District of Red Paal with 80 respondents selected by purposive sampling. Data collection through interviews, observation, measurement of cholinesterase levels, measurement of hemoglobin levels and measurement of the amount of erythrocytes in the blood. Data analysis using chi-square test and logistic regression.&#x0D; Results: Based on the results of bivariate analysis, it was found that there were 3 independent variables related to hemoglobin levels, namely work period (p value = 0.005), frequency of spraying (p value = 0.007), pesticide dose (p value = 0.003, and no independent variables related to the number of erythrocytes.&#x0D; Conclusion: Based on research from 80 farmers who examined hemoglobin levels in the blood there were 73.8% of farmers who experienced abnormal hemoglobin levels, and 26.3% of farmers who experienced normal hemoglobin levels, and from 80 farmers who examined the amount of erythrocytes in blood is 22.5% of farmers who have an abnormal number of erythrocytes, and 77.5% of farmers have normal erythrocyte counts.

Engagement and Education of the Sickle Cell Disease Community: What Sickle Cell Disease Patients and Parents Want to Know about CRISPR Genome Editing

Introduction:One of the first therapeutic targets of CRISPR genome editing will likely be sickle cell disease (SCD), the most commonly inherited blood disorder in the U.S.SCD affects 100,000 individuals in the U.S. and more than 250 million individuals globally. Two targets of genome editing in SCD currently under investigation are HBB and BCL11A. The first approach (HBB) focuses on correcting the Glu6Val mutation in patient-derived stem and progenitor cells to promote differentiation into normal erythrocytes. The second approach involves disrupting BCL11A, a repressor of fetal hemoglobin. While preliminary results from research using both mechanisms show promise, risks related to insertional mutagenesis and host immune responses remain. As the research advances, limited knowledge exists regarding SCD stakeholders' (patients, parents and physicians) understanding of genome editing and their educational needs.Methods:We conducted a mixed methods study, which included a CRISPR genome editing educational video, pre- and post-video surveys, and fifteen, moderated focus groups: 6 patient groups, 6 parent groups, and 3 physician groups. The surveys included an assessment of participants' knowledge of CRISPR gene editing and the Genetic Literacy and Comprehension (GLAC) instrument. Eligible participants were patients with a diagnosis of SCD, parents of a child with SCD, and physicians who have delivered care to at least five adult or pediatric patients living with SCD for a minimum of 12 months. Differences between the pre- and post-video score of CRISPR technology knowledge were assessed using paired t-tests. NVivo 11 qualitative software tool was used to assist in data analysis.Results:For each of the eight complex genetic terms in the GLAC, patients and parents were asked to rank their familiarity on a scale from 1-7, and to complete fill-in-the-blank, multiple choice questions aimed at assessing their understanding of the term. Baseline genetic literacy levels were high for both stakeholder groups. Patients and parents had an average familiarity score of 6.34, and 6.88, respectively. 89% of both groups scored >70% on the fill-in-the-blank questions. As shown in Figure 1, the educational video tool was effective at increasing CRISPR genome editing knowledge among patients and parents - the average CRISPR genome editing knowledge scores of both groups increased by over two points (p<0.0001 and p=0.05, respectively). Prior to viewing the video, patients and parents had lower CRISPR genome-editing knowledge scores than physicians (p<0.0002 and p<0.0002, respectively). However, after viewing the video, there were no significant differences in scores between patients, parents, and physicians. Several qualitative themes emerged related to patients' and parents' interest in understanding CRISPR genome editing. These themes included the need for easily accessible scientific information regarding: 1) CRISPR's mechanism of action; 2) potential adverse reactions to CRISPR; 3) effects of genome editing on future generations; and 4) burden of participating in a genome editing clinical trial.Discussion:After watching the video tool, patients' and parents' understanding of CRISPR genome editing improved significantly. This suggests that video education tools may be an effective method to educate individuals living with SCD and parents about genome editing. Furthermore, this study reveals patients and parents were not only familiar with complex genetic terms, but also correctly defined them, suggesting a good baseline genetic literacy. The themes indicate the importance of keeping patients and parents informed and supporting their desires to be educated about the applications of genome editing technology which will further the advancement of clinical trials. DisclosuresNo relevant conflicts of interest to declare.

Regulation of Mature Erythrocyte Proteasomal Machinery Under Hypoxia

Proteasome machinery is a conserved cellular component to maintain normal protein homeostasis. Hypoxia is well known to control hypoxia inducible factor levels by proteasomal machinery in nucleated cells. However, the specific targets and regulation of proteasomal machinery in non-nucleated mature erythrocytes under hypoxia remain poorly understood. To determine if hypoxia regulates erythrocyte proteasomal machinery, we conducted Western blot to detect total ubiquitinated and K48 specific ubiquitinated proteins on the erythrocyte membrane in both human and mice with or without sickle cell disease (SCD), a hemolytic genetic disease with a high mortality, morbidity and frequently facing hypoxia. We found that ubiquitinated, especially K48 specific ubiquitinated proteins were significantly accumulated in both SCD Berkeley mice and humans compared to WT mice and normal controls, indicating that proteasomal machinery is impaired in SCD. Next, to determine specific ubiquitinated proteins accumulated on the membrane of human sickle erythrocytes (sRBC), we conducted immunoprecipitation of sRBC membrane proteins with total ubiquitin antibody followed by an robust and nonbiased proteomic profiling. We found significant accumulation of several categories of ubiquitinated proteins on human mature sRBC membrane, including cytoskeleton proteins (Spectrin, Actin, Ankryin), glycolytic enzymes (GAPDH, 2,3-BPG mutase, Pyruvate Kinase, G6PD), transporters (Band3, large neutral AA transporter, calcium transporter, ENT1), reactive oxygen species (ROS) related enzyme (catalase), components of proteasome machinery [E2, E3 ligases, and valosin-containing protein (p97)]. Mechanistically, we revealed that the impaired proteasomal machinery found in mature sRBC was due to the blockage of trafficking of p97 bound ubiquitinated-proteins from membrane to cytosolic proteasome. As such, inhibition of p97 by CB-5083 or proteasome by MG132 led to further induction of hypoxia-induced ubiquitinated membrane proteins and sickling in cultured human sRBC. Given the fact that sphingosine-1-phosphate (S1P) contributes to sickling by binding with deoxygenated sickle Hb (deoxy-HbS), triggering deoxy-HbS anchoring membrane and releasing glycolytic enzymes, we immediately hypothesized that S1P may be involved in proteasomal machinery by regulating trafficking of p97-bound ubiquitinated proteins from membrane to cytosol in sRBC. To test this intriguing possibility, we generated SCD/Sphk1-/- mouse. Intriguingly, we found that the genetic deletion of SphK1 attenuated impaired proteasomal machinery in sRBC with less accumulation of p97 and ubiquitinated proteins on sRBC membrane, indicating that elevated S1P is detrimental in sRBC by inducing accumulation of p97 and ubiquitinated proteins on the membrane. Moreover, to determine if S1P-regulated p97 trafficking from membrane to the cytosol is unique to sRBC, we exposed wild type and SphK1-/- mice to 8% hypoxia up to 72 hours. In contrast to sRBC, we found that genetic deletion of SphK1 abolished p97 trafficking from membrane to cytosol in normal erythrocytes under hypoxia. Finally, we conducted in vitro proof-of-principle genetic studies to determine if S1P directly involves in translocation of membrane anchored p97 to cytosol using inverted ghost membrane (IGM). We demonstrated that S1P treatment induces deoxy-HbA translocation from cytosol to membrane and in turn restoring p97 release from membrane to the cytosol in IGM isolated from SphK1-/- mice only under hypoxia but not normoxia. Thus, we have provided both human and mouse genetic evidence supporting a working model: in normal individuals under hypoxia, S1P is a key factor regulating the efficient proteasomal machinery by binding deoxy-HbA, promoting deoxy-HbA anchoring membrane and in turn triggering release of p97-ubiqutinated proteins to cytosol for its degradation. With mutation in HbS, S1P promotes deoxy-HbS anchoring the membrane and forms polymers, thus blocks membrane bound p97 release and impairs proteasomal machinery in sRBC. Overall, our findings identified that S1P is a missing key component of proteasome machinery by its differential mechanism regulating p97 trafficking from membrane to cytosol in normal erythrocyte physiology under hypoxia and the pathophysiology of SCD that open up new and differential therapies for the SCD and normal individuals facing hypoxia. DisclosuresNo relevant conflicts of interest to declare.

Transcripts of antibacterial peptides in chicken erythrocytes infected with Marek\u2019s disease virus

BackgroundChicken erythrocytes are involved in immunity through binding of toll-like receptors (TLRs) with their ligands to activate downstream signaling and lead to cytokine production in erythrocytes. Some avian β-defensins (AvBDs) are constitutively expressed in tissues and some others can be induced by various bacteria and viruses. However, the expression of AvBDs in erythrocytes has not yet been studied extensively.ResultsThe transcripts of eight AvBDs (AvBD1 to AvBD7, and AvBD9) and liver-expressed antimicrobial peptide-2 (LEAP-2) were found in normal chicken erythrocytes. The expression levels of AvBD2, 4 and 7 were significantly increased (P < 0.01), whereas the levels of AvBD1, 6 and 9 were significantly decreased (P < 0.01) after Marek’s disease virus (MDV) infection. The mRNA expression level of LEAP-2 was not significantly changed after MDV infection. Highest viral nucleic acid (VNA) of MDV in the feather tips among the tested time points was found at 14 days post-infection (d.p.i.). In addition, 35 MD5-related gene segments were detected in the erythrocytes at 14 d.p.i. by transcriptome sequencing.ConclusionsThese results suggest that the AvBDs in chicken erythrocytes may participate in MDV-induced host immune responses.

Morphological changes in the ovarian carcinoma cells of Wistar rats induced by chemotherapy with cisplatin and dioxadet.

The development of new express methods for the analysis of the efficacy of anti-cancer therapy on the cellular level is highly desirable for the analysis of chemotherapeutic agent performance. In this paper we suggest the use of parameters of cell morphology determined by holographic microscopy and tomography for the effective label free quantitative analysis of cell viability under antitumor chemotherapy and thus of cytostatic agent efficacy. As shown, measured phase shifts and cell morphology change dramatically as a result of chemotherapy and depend strongly on the cell type and agent applied. Experimentally, a comparative analysis of the antitumor efficacy of the two cytostatics, cisplatin and dioxadet, that are commonly used for chemotherapy of disseminated ovarian carcinoma has been performed. The experiments were carried out on the Wistar rat model. An essential difference in the morphology of cells, both normal (erythrocytes) and cancerous, present in ascitic fluid taken from the non-treated group of rats and the groups treated with either dioxadet or cisplatin, has been observed. The results obtained can be interpreted as an indication of the antitumor performance of both cytostatics at the cellular level and as a demonstration of the higher efficacy of therapy with dioxadet as compared to that with cisplatin. Differences in cell morphology are suggested to be applied as quantitative markers of cell viability and cytostatic agent efficacy. The conclusions made are supported by a comparison with the results of recent experiments based on survival rates of laboratory animals treated with these agents..

Microscopic Monitoring of Erythrocytes Deformation under Different Shear Stresses Using Computerized Cone and Plate Flow Chamber: Analytical Study of Normal Erythrocytes and Iron Deficiency Anemia.

Erythrocytes deformation is one of the exciting properties of erythrocytes. It is still under investigation by many of the researchers in different disciplines. The clinicians and researchers are still looking for a simple and efficient method to monitor and tracking the erythrocytes deformation. This research article represented a microscopic technique that could be a helpful tool in evaluation and studying of erythrocytes deformation under different shear stresses. This technique was used to compare the deformation of normal erythrocytes and iron deficiency anemia. Elongation index was calculated and used in the quantitative analysis of erythrocytes deformation. It was found that the deformability of normal erythrocytes was higher than that of iron deficiency anemia. Normal erythrocytes and iron deficiency anemia showed strong negative correlations with the mean cell volume and the mean cell hemoglobin concentration under different shear stresses. This study introduces more understanding of the erythrocytes deformation by using a simple microscopic technique. The elongation index could be used as a measurable parameter to evaluate the deformability of the erythrocyte in normal and abnormal cases.

Absolute proteome quantification of highly purified populations of circulating reticulocytes and mature erythrocytes

AbstractReticulocytes produced in the bone marrow undergo maturation in the bloodstream to give rise to erythrocytes. Although the proteome of circulating red cells has been the subject of several reports, the cellular populations used for these studies were never completely devoid of reticulocytes. In our current study, we used highly purified erythrocyte and reticulocyte populations to quantify the absolute expression levels of the proteins in each cell population. Erythrocytes and reticulocytes were purified in a multistep process involving cellulose chromatography, Percoll gradient centrifugation, and fluorescence cell sorting after thiazole orange labeling. Proteins were analyzed by mass spectrometry from whole cells and erythrocyte plasma membrane (ghosts), leading to the identification and quantification of 2077 proteins, including 654 that were reticulocyte-specific. Absolute quantifications of these proteins were made using the mean corpuscular hemoglobin content of the cells as a standard. For each protein, we calculated the percentage loss during the terminal stages of reticulocyte maturation and the percentage of association with the plasma membrane. In addition, we used modified adenosine triphosphate and adenosine diphosphate molecules that enable the transfer of a biotin molecule to the catalytic sites of kinases to isolate active kinases in the erythrocytes and determined the absolute expression of 75 protein kinases and the modification of their expression during reticulocyte maturation. Our findings represent the first absolute quantification of proteins that are specifically expressed in normal erythrocytes with no detectable contamination by reticulocytes. Our findings thus represent a reference database for the future proteomic analysis of pathological erythrocytes.

Antioxidant properties and membrane stabilization effects of methanol extract of Mucuna pruriens leaves on normal and sickle erythrocytes

This study was aimed at investigating the antioxidant properties and membrane stabilization effects of Mucuna pruriens leaves on sickle erythrocyte as a possible means of sickle cell disease management.Pulverized plant material was extracted with methanol, filtered and concentrated at reduced pressure with a rotary evaporator. Phytochemical analysis and antioxidant studies of the extract were carried out using standard methods. Blood samples of volunteer sickle cell patients and healthy individuals used in the study were collected from the University of Nigeria Medical Centre and University campus community, Nsukka respectively. The genotypes of the individuals were confirmed by cellulose acetate paper electrophoresis. Water induced haemolysis of human red blood cell was used to assess membrane stabilization of the erythrocytes. Phytochemical result of the extract showed the presence of alkaloids, flavonoids, tannins, proteins, terpenoids, saponins, cardiac glycosides and anthraquinones. Antioxidant vitamins C and E were present in concentrations of 495.36 mg/100 g and 101.03 mg/100 g respectively. The percentage (%) scavenging activity of the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and hydroxyl radical by the extract was significant. The extract exhibited membrane stabilization on both normal and sickle erythrocytes. The percentage (%) inhibition of haemolysis by the extract in both normal and sickle erythrocytes at different concentrations of 100, 200, 400, 600 and 800 μg/ml were significant and concentration dependent. We conclude that M. pruriens leaves have antioxidant properties and erythrocyte membrane stabilizing potentials and could be recommended for use in the management of patients with sickle cell anaemia.

Improved efficacy of doxycycline in liposomes against Plasmodium falciparum in culture and Plasmodium berghei infection in mice.

The rate at which Plasmodium falciparum is developing resistance to clinically used antimalarial drugs is alarming. Therefore, there is a compelling need to develop an efficient drug delivery system to improve the efficacy of existing antimalarial agents and circumvent drug resistance. Here, we report the antibacterial drug doxycycline (DOXY) in liposomal formulations exhibits enhanced antiplasmodial activity against blood stage forms of P. falciparum (3D7) in culture and established Plasmodium berghei NK-65 infection in murine model. Parasite killing on blood stage forms in culture was determined by a radiolabeled [3H] hypoxanthine incorporation assay and infected erythrocytes stained with Giemsa were counted using microscopy in vivo. The 50% inhibitory concentration (IC50) of DOXY-stearylamine liposome (IC50 0.36 μM) and DOXY-SPC:Chol-liposome (IC50 0.85 μM) exhibited marked growth inhibition of parasites compared with free DOXY (IC50 14 μM), with minimal toxicity to normal erythrocytes. Administration of polyethylene glycol distearoyl phosphatidylethanolamine-methoxy-polyethylene glycol2000 (DSPE-mPEG-2000) coated liposomes loaded with DOXY at 2.5 mg/kg per day resulted in efficacious killing of blood parasites with improved survival in mice relative to the free drug in both chloroquine sensitive and resistant strains of P. berghei infection. This is the first report to demonstrate that DOXY in liposomal system has immense chemotherapeutic potential against plasmodial infections at lower dosages.

Profil Kadar Hemoglobin dan Indeks Eritrosit pada Perokok Aktif di Kelurahan Tanjung Pinang Kota Palangka Raya

Active smokers are people who are smoking continuously. The population of active smokers in Central Kalimantan was 26,5% in 2013. Cigarettes contain many toxic and addictive chemicals. The presence of tar and free radicals from cigarette smoke can cause erythrocyte hemolysis. In addition, the content of cigarette smoke can also cause an increase in hemoglobin levels that mediated by exposure to Carbon monoxide (CO). CO binds avidly to hemoglobin to form carboxyhemoglobin (HbCO). This study aims to give a description of hemoglobin level and erythrocyte indices in active smokers in Kelurahan Tanjung Pinang, Palangka Raya. The descriptive observational method with the cross-sectional design was used in this study. The sample obtained by 28 people with purposive sampling technique with criteria such as active smokers, men, working as farmers, laborers or fish farmers and not alcoholic. Examination of hemoglobin level and erythrocytes indices was done by a hematology analyzer. The result showed that 4 people (14,2%) had low hemoglobin level and 4 people (85,7%) was normal. Based on erythrocytes indices, 2 people (7,1%) with hypochromic microcytic anemia (low MCV, low MCH, and normal MCHC) and also low hemoglobin level. While 26 people (92,8%) had normal erythrocyte indices.

A Mutation Outside the Dimerization Domain Causing Atypical STING-Associated Vasculopathy With Onset in Infancy.

Mutations in the gene encoding stimulator of interferon genes (STING) underlie a type I interferon (IFN) associated disease, STING-associated vasculopathy with onset in infancy (SAVI). Patients suffer cutaneous vasculopathy and interstitial lung disease, but are not known to suffer life-threatening infection. We describe a child who presented with Pneumocystis jirovecii pneumonia in early life, from which he recovered. He went on to suffer failure to thrive, developmental delay, livedo reticularis, and vesicular rash, but without cutaneous vasculitis, and with normal C-reactive protein and erythrocyte sedimentation rates. At 3 years of age, he developed life-threatening pulmonary hypertension. Whole genome sequencing (WGS) was performed using the Illumina HiSeqX10 platform and the Seave platform was used for bioinformatic analysis. mRNA expression of IFN-stimulated genes and inflammatory cytokines from peripheral blood mononuclear cells was determined by quantitative polymerase chain reaction. Luciferase assay was used to model IFNβ and NF-κB activity in vitro. WGS revealed a de novo mutation p.Arg284Ser in STING at an amino acid previously associated with SAVI. Although this mutation did not fall in the dimerization domain (DD), mRNA analysis revealed constitutive IFN-gene activation consistent with an interferonopathy, which correlated to STING activation in vitro. The patient was treated with corticosteroids and the JAK inhibitor Ruxolitinib, resulting in a rapid improvement of pulmonary hypertension, general well-being, and resolution of the IFN gene signature. However, he did go on to evolve a nasal septal erosion suggesting incomplete control of disease. This case provides molecular evidence to support the p.Arg284Ser variant in STING exerting pathogenicity through a gain-of-function mechanism. The lack of cutaneous vasculitis or elevated systemic inflammatory markers, and the occurrence of an opportunistic infection are notable, and raise the possibility that variants outside the STING DD may potentially manifest with an atypical SAVI phenotype. Nevertheless, there was an objective clinical improvement in response to JAK inhibition.