Normal Erythrocytes Research Articles

Phytochemical Analysis, Total Phenolic Content, Hemolytic and Anti-hemolytic Activities of Centaurea iberica (Asteraceae)

Several plants contain chemical substances that have various biological activities, but they also probably cause harmful effects on cellular membranes. This work deals with the Centaurea iberica (Iberian starthistle) plant. The objective of this study is to identify the main chemical groups in the methanolic extract 80% of the plant, determine its total phenolic content, evaluate its toxic effect on human erythrocytes, and also assess its activity in the protection of normal and glucose-6-phosphate dehydrogenase (G6PD) enzyme-deficient human erythrocytes membranes against oxidative hemolysis. Phytochemical analysis revealed that C. iberica extract contains several bioactive compounds. The results also showed that the plant extract contained good level of total phenolic compounds. The extract exhibited a low hemolytic effect. It also showed excellent activity in the protection of normal and G6PD-deficient human erythrocytes against oxidative hemolysis. KEYWORDS Erythrocytes, G6PD, Iberian starthistle, methanolic extract, toxic effect, oxidative hemolysis

Morphological Factors and Forces Acting in Normal and Sickled Erythrocytes in Sickle Cell Anemia: A Mathematical Model

We carried out a theoretical study that considers the morphological factors and forces acting on a normal and a sickled blood cell. Sickle cell disease is associated with vaso-occlusion which creates blood flow crises as a result of loss in shape memory of the normal red blood cell. Certain forces and chemical compositions acting on the cell could retain the shape memory. The problem of a second order partial differential equation modeled was solved to get the forces and chemical composition required to restore the sickled red blood cells back to its original shape. The results gotten showed that, increase in chemical reaction increased the RBC growth by creating a stretching force that causes the sickle cell to regain elasticity with improved oxygen.

Gas6/Axl signaling promotes hematoma resolution and motivates protective microglial responses after intracerebral hemorrhage in mice

BackgroundIntracerebral hemorrhage (ICH) stands out as the most fatal subtype of stroke, currently devoid of effective therapy. Recent research underscores the significance of Axl and its ligand growth arrest-specific 6 (Gas6) in normal brain function and a spectrum of neurological disorders, including ICH. This study is designed to delve into the role of Gas6/Axl signaling in facilitating hematoma clearance and neuroinflammation resolution following ICH. MethodsAdult male C57BL/6 mice were randomly assigned to sham and ICH groups. ICH was induced by intrastriatal injection of autologous arterial blood. Recombinant mouse Gas6 (rmGas6) was administered intracerebroventricularly 30 min after ICH. Virus-induced knockdown of Axl or R428 (a selective inhibitor of Axl) treatment was administrated before ICH induction to investigate the protective mechanisms. Molecular changes were assessed using western blot, enzyme-linked immunosorbent assay and immunohistochemistry. Coronal brain slices, brain water content and neurobehavioral tests were employed to evaluate histological and neurofunctional outcomes, respectively. Primary glia cultures and erythrophagocytosis assays were applied for mechanistic studies. ResultsThe expression of Axl increased at 12 h after ICH, peaking on day 3. Gas6 expression did not remarkably changed until day 3 post-ICH. Early administration of rmGas6 following ICH significantly reduced hematoma volume, mitigated brain edema, and restored neurological function. Both Axl-knockdown and Axl inhibitor treatment abolished the neuroprotection of exogenous Gas6 in ICH. In vitro studies demonstrated that microglia exhibited higher capacity for phagocytosing eryptotic erythrocytes compared to normal erythrocytes, a process reversed by blocking the externalized phosphatidylserine on eryptotic erythrocytes. The erythrophagocytosis by microglia was Axl-mediated and Gas6-dependent. Augmentation of Gas6/Axl signaling attenuated neuroinflammation and drove microglia towards pro-resolving phenotype. ConclusionsThis study demonstrated the beneficial effects of recombinant Gas6 on hematoma resolution, alleviation of neuroinflammation, and neurofunctional recovery in an animal model of ICH. These effects were primarily mediated by the phagocytotic role of Axl expressed on microglia.

Temporal arteritis with normal erythrocyte sedimentation rate and C-reactive protein at presentation

Temporal arteritis with normal erythrocyte sedimentation rate and C-reactive protein at presentation

NDV targets the invasion pathway in malaria parasite through cell surface sialic acid interaction.

Merozoites utilize sialic acids on the red blood cell (RBC) cell surface to rapidly adhere to and invade the RBCs. Newcastle disease virus (NDV) displays a strong affinity toward membrane-bound sialic acids. Incubation of NDV with the malaria parasites dose-dependently reduces its cellular viability. The antiplasmodial activity of NDV is specific, as incubation with Japanese encephalitis virus, duck enteritis virus, infectious bronchitis virus, and influenza virus did not affect the parasite propagation. Interestingly, NDV is reducing more than 80% invasion when RBCs are pretreated with the virus. Removal of the RBC surface proteins or the NDV coat proteins results in disruption of the virus binding to RBC. It suggests the involvement of specific protein: ligand interaction in virus binding. We established that the virus engages with the parasitized RBCs (PRBCs) through its hemagglutinin neuraminidase (HN) protein by recognizing sialic acid-containing glycoproteins on the cell surface. Blocking of the HN protein with free sialic acid or anti-HN antibodies abolished the virus binding as well as its ability to reduce parasite growth. Interestingly, the purified HN from the virus alone could inhibit the parasite's growth in a dose-dependent manner. NDV binds strongly to knobless murine parasite strain Plasmodium yoelii and restricted the parasite growth in mice. Furthermore, the virus was found to preferentially target the PRBCs compared to normal erythrocytes. Immunolocalization studies reveal that NDV is localized on the plasma membrane as well as weakly inside the PRBC. NDV causes neither any infection nor aggregation of the human RBCs. Our findings suggest that NDV is a potential candidate for developing targeted drug delivery platforms for the Plasmodium-infected RBCs.

Repair of inflammatory celiac artery aneurysm presenting with insidious onset of abdominal pain: A case report

BackgroundVisceral artery aneurysms (VAAs) are rare but potentially devastating given the high mortality associated with rupture. Current Society of Vascular Surgery guidelines recommend repair of these at 2.5 cm. Surgical intervention includes endovascular embolization and/or stenting versus open revascularization. This case presents a patient with a large celiac artery aneurysm (CAA) who underwent open ligation of the celiac artery at the origin. Splenectomy was performed given the need for ligation of the splenic artery. He had a replaced right hepatic artery from the SMA; thus, no revascularization was needed. Ultimately, the aneurysm was found to be inflammatory in etiology. Case reportA 41-year old male with history of seizure disorder, smoking, and newly diagnosed hypertension presented with abdominal pain for six months. His-laboratory work was significant for a leukocytosis of 17, 740, mildly elevated C- Reactive Protein (CRP) at 1.63, and normal erythrocyte sedimentation rate (ESR). CT angiogram revealed a 5.2 × 4.2 cm celiac artery aneurysm with stranding. Given its appearance and concern for infection, he underwent emergent laparotomy and ligation. Blood cultures and intraoperative tissues revealed no microorganism growth. Pathology was remarkable only for periarterial abscess with acute and chronic inflammation and focal tissue necrosis, compatible with aneurysm. ConclusionThis case report presents a rare inflammatory CAA, which presented with six months of abdominal pain, and its subsequent management with open intervention. In this case, his vessel anatomy was favorable for open repair with ligation of the celiac axis due to the presence of a replaced right hepatic artery. This also highlights the importance of anatomic vessel variants when making surgical decisions.

Development of clinical and laboratory biomarkers in an international cohort of 428 children with lupus nephritis.

Lupus nephritis (LN) is a very severe manifestation of lupus. There is no consensus on which treatment goals should be achieved to protect kidney function in children with LN. We retrospectively analyzed trends of commonly used laboratory biomarkers of 428 patients (≤ 18years old) with biopsy-proven LN class ≥ III. We compared data of patients who developed stable kidney remission from 6 to 24months with those who did not. Twenty-five percent of patients maintained kidney stable remission while 75% did not. More patients with stable kidney remission showed normal hemoglobin and erythrocyte sedimentation rate from 6 to 24months compared to the group without stable kidney remission. eGFR ≥ 90ml/min/1.73m2 at onset predicted the development of stable kidney remission (93.8%) compared to 64.7% in those without stable remission (P < 0.00001). At diagnosis, 5.9% and 20.2% of the patients showed no proteinuria in the group with and without stable kidney remission, respectively (P = 0.0001). dsDNA antibodies decreased from onset of treatment mainly during the first 3months in all groups, but more than 50% of all patients in both groups never normalized after 6months. Complement C3 and C4 increased mainly in the first 3months in all patients without any significant difference. Normal eGFR and the absence of proteinuria at onset were predictors of stable kidney remission. Significantly more children showed normal levels of Hb and erythrocyte sedimentation rate (ESR) from 6 to 24months in the group with stable kidney remission.

Upregulation of ATP6V0D2 benefits intracellular survival of Leishmania donovani in erythrocytes-engulfing macrophages.

Visceral leishmaniasis (VL) is the most severe type of leishmaniasis which is caused by infection of Leishmania donovani complex. In the BALB/c mouse model of VL, multinucleated giant cells (MGCs) with heavy parasite infection consist of the largest population of hemophagocytes in the spleen of L. donovani-infected mice, indicating that MGCs provide the parasites a circumstance beneficial for their survival. Although ATP6V0D2 is a demonstrated factor inducing the formation of hemophagocytic MGCs during L. donovani infection, functions of this protein in shaping the infection outcome in macrophages remain unclear. Here we evaluated the influence of upregulated ATP6V0D2 on intracellular survival of the parasites. L. donovani infection-induced hemophagocytosis of normal erythrocytes by macrophages was suppressed by RNAi-based knockdown of Atp6v0d2. The knockdown of Atp6v0d2 did not improve the survival of amastigotes within macrophages when the cells were cultured in the absence of erythrocytes. On the other hand, reduced intracellular survival of amastigotes in macrophages by the knockdown was observed when macrophages were supplemented with antibody-opsonized erythrocytes before infection. There, increase in cytosolic labile iron pool was observed in the L. donovani-infected knocked-down macrophages. It suggests that ATP6V0D2 plays roles not only in upregulation of hemophagocytosis but also in iron trafficking within L. donovani-infected macrophages. Superior access to iron in macrophages may be how the upregulated expression of the molecule brings benefit to Leishmania for their intracellular survival in the presence of erythrocytes.

Novel betulin derivatives as multidrug reversal agents targeting P-glycoprotein

Chemotherapy is a powerful means of cancer treatment but its efficacy is compromised by the emergence of multidrug resistance (MDR), mainly linked to the efflux transporter ABCB1/P-glycoprotein (P-gp). Based on the chemical structure of betulin, identified in our previous work as an effective modulator of the P-gp function, a series of analogs were designed, synthesized and evaluated as a source of novel inhibitors. Compounds 6g and 6i inhibited rhodamine 123 efflux in the P-gp overexpressed leukemia cells, K562/Dox, at concentrations of 0.19 µM and 0.39 µM, respectively, and increased the intracellular accumulation of doxorubicin at the submicromolar concentration of 0.098 µM. Compounds 6g and 6i were able to restore the sensitivity of K562/Dox to Dox at 0.024 µM and 0.19 µM, respectively. Structure–activity relationship analysis and molecular modeling revealed important information about the structural features conferring activity. All the active compounds fitted in a specific region involving mainly transmembrane helices (TMH) 4–6 from one homologous half and TMH 7 and 12 from the other, also showing close contacts with TMH 6 and 12. Compounds that bound preferentially to another region were inactive, regardless of their free energy of binding. It should be noted that compounds 6g and 6i were devoid of toxic effects against peripheral blood mononuclear normal cells and erythrocytes. The data obtained indicates that both compounds might be proposed as scaffolds for obtaining promising P-gp inhibitors for overcoming MDR.

Defining Hereditary Pyropoikilocytosis (HPP) in the Molecular Diagnostic Era

Misconceptions continue in distinguishing hereditary elliptocytosis (HE) with hemolysis in early perinatal period and hereditary pyropoikilocytosis (HPP). In 1975, Zarkowsky and colleagues described a group of children with transfusion dependent congenital hemolytic anemia with the unique morphology of “microspherocytes, measuring 2-3 microns in diameter, and cells with blunted projections or triangular in shape”, a morphology resembling the heat induced damage to normal erythrocytes - hence the name hereditary pyropoikilocytosis (HPP). Subsequently, similar cases were described with or without demonstrable heat sensitivity (Ravindranath &amp; Johnson, 1985). Regardless, the common denominators were 1) transfusion dependency early in infancy - resolved with splenectomy, 2) severe microcytosis, often &amp;lt;60 fL, and 3) a decrease in membrane bound spectrin (decreased spectrin/band3 ratio). Family studies showed normal RBC morphology in both parents (Ravindranath &amp; Johnson, 1985) or in some cases, one parent with HE (Zarkowsky et al., 1975). Protein chemistry studies showed decreased tetramer formation and tryptic digests showed mutations within the n-terminal alpha 1 domain of alpha spectrin - the dimer/dimer interaction site. Thus, homozygosity or double heterozygosity of mutations in the dimer/dimer interaction site of alpha spectrin (alpha 1 domain of alpha spectrin) defined HPP while classical HE cases showed heterozygous mutations in the alpha 1 domain. A fact unexplained was how this results in spectrin deficiency, specifically, alpha spectrin deficiency (Ravindranath &amp; Johnson, 1985). Using the molecular analytical tool AnemiaID (courtesy of Agios Pharmaceuticals, Inc.), we show that HPP is defined by the co-inheritance of two SPTA1 mutations and Alpha-LeLy polymorphism ( αLeLy rs28525570; SPTA1[c.5572C&amp;gt;G;c.6531-12C&amp;gt;T]) in one or both alleles, thus explaining the increased propensity for red cell fragmentation and the spectrin deficiency noted on acrylamide gels [Table: Clinical and molecular characteristics of our cases]. Of note, homozygosity for αLeLy polymorphism and one SPTA1 variant did not result in clinical HPP phenotype [HE-2 in Table]. At a clinical laboratory level, the extreme microcytosis in HPP is evident in the markedly diminished band3 content on eosin-5'-maleimide (EMA) binding test- mean channel fluorescence (MCF) of &amp;lt;275.0 (control: 473.0-549.0) while HE (including neonatal HE with hemolysis) show a double peak pattern in EMA (the minor peak representing the fragmented cells while the major peak had MCF higher than control MCF) and the characteristic trapezoid pattern on osmotic gradient ektacytometry (not shown).

Adhesion of Immunoglobulins to Band3 Promotes Increased Erythrocyte Sedimentation Rate in Multiple Myeloma

An increase in ESR can be seen in approximately 85% of patients with MM [1]. Therefore, ESR is often used as one of the sensitive indicators of active MM [1]. It is now widely accepted that the increase in ESR is related to the adhesion of immunoglobulins secreted by myeloma cells to erythrocytes [2, 3]. However, this explanation is descriptive and speculative and has not been confirmed by experimental data [3]. We applied peripheral blood erythrocyte samples from MM patients and used immunofluorescence/immunoprecipitation tandem protein analysis/molecular docking to determine the molecular mechanism by which all types of MM immunoglobulins can target and adhere to erythrocyte membrane Band3. This result, not only helps to explain the molecular mechanism of ESR acceleration in MM, but will also advance our understanding of the pathophysiologic changes that occur in ESR. At the same time, it may provide a new way to target protection of normal erythrocytes and targeted removal of erythrocyte-bound immunoglobulins.

A Phase I Clinical Trial of CLL-1 CAR-T Cells for the Treatment of Relapsed/Refractory Acute Myeloid Leukemia in Adults

Background: Relapsed/refractory acute myeloid leukemia (r/r AML) has a poor prognosis, and most patients fail standard chemotherapy and hematopoietic stem cell transplantation or relapse after treatment. We previously reported and evaluated CLL-1-targeted chimeric antigen receptor T-cell therapy in adult r/r AML patients, demonstrating for the first time the efficacy and tolerable safety of CLL-1 CAR-T cell therapy in adult r/r AML patients. On this basis, we expanded the data volume and continued to evaluate the safety and efficacy of CLL-1 CAR-T in AML with dose escalation. Methods: This is a single-center, phase I, dose-escalation trial of CLL-1 CAR-T cell in adult patients with r/r AML (ChiCTR2000041054）admitted to Tianjin First Central Hospital from January 2021 to June 2023. All patients received a lymphodepleting chemotherapy with cyclophosphamide (500mg/m2) and fludarabine (30mg/m2) 3 days before CAR-T cell infusion, followed by 4 incremental doses of CAR-T cells: dose level (DL)1 (0.5×10^6/kg），DL2（1×10^6/kg），DL3（1.5×10^6/kg）and DL4（2×10^6/kg）CAR-T cells. The primary objective of the trial was to evaluate safety and tolerability, and to determine the recommended Phase 2 dose. Key secondary objectives included the assessment of overall response rate, complete response rates as well as progression free survival and overall survival. Results: Thirty patients were enrolled and treated in the dose-escalation phase of the study. The median age of the patients was 38 years (range 18-73), and the median number of prior lines of treatment was 4(2-10), 5(16.67%) patients progressed from MDS to AML, 13(43.33%) patients relapsed after treatment, of which 8(26.67%) patients had previously received HSCT. There were 9, 9, 6 and 6 patients receiving DL1, DL2, DL3 and DL4, respectively. All patients experienced cytokine release syndrome (CRS) (100%): grade 0-2(n=18), grade 3(n=11), grade 4(n=1), and only one patient (3.33%) experienced grade 4 neurotoxicity (NTX), which improved after plasma exchange. In addition, all patients experienced hematological toxicity, with 29 patients (96.67%) developing grade 3/4 granulocytopenia, 28 patients (93.33%) developing grade 3/4 anemia, and 30 patients (100%) developing thrombocytopenia. Patients who received haploidentical bridge transplantation after infusion had normal granulocyte, erythrocyte, and platelet transplantation and did not develop serious infection, suggesting that when severe agranulocytosis occurs after CAR-T infusion, bridge transplantation can reverse this toxicity (Table 1). Efficacy evaluation showed that 73% of patients responded to the infusion, with 12(40%) patients achieving MRD-CR/CRi and 10(33.33%) patients achieving MRD+CR/CRi, for an overall response rate (ORR) of 73.33% (Figure 1). All patients who underwent bridging haploidentical HSCT after CAR-T cell infusion achieved MRD-CR/CRi, and the median time to transplantation was 0.7 months (0.3-1.13). The median OS rate of patients after transplantation was higher than that of patients without transplantation after infusion (P=0.015), as was PFS (p=0.044). The maximum tolerated dose was not determined as no dose-limiting toxic effects were observed. Responses were observed at all dose levels, with a median follow-up of 132 days (range 15-914 days). Patients had a median PFS of 300 days (95% CI, 191.320 to 408.680) and a median OS of 348 days (95% CI, 234.011 to 461.989). The expansion of CLL-1CAR-T cells was tracked by flow cytometry, and the median peak of CAR-T expansion occurred at day 11 (range 2-15) after infusion. When comparing the peak CAR-T expression in CR/CRi and NR patients, the proportion of peak CAR-T cells was significantly higher in CR/CRi patients. Conclusion: CLL-1 is a potential therapeutic target in AML. The proportion of patients who developed severe CRS was low and only 1 patient experienced NTX. Although severe agranulocytosis may occur, patients receiving CLL-1CAR-T therapy may have the opportunity to achieve CR/CRi prior to transplantation, which may reduce the risk of relapse and mortality, and prolong patient survival with encouraging results. Due to the small sample size, no relevant dose-response relationship was observed, and the trial is currently enrolling more cases and will be progressively expanded to Phase II clinical trials.

Register-based observational study of associations between inflammatory remission, formal treatment targets and the use of disease-modifying antirheumatic drugs among patients with early rheumatoid arthritis

ObjectiveTo assess associations between inflammatory remission, formal treatment targets and the likelihood of starting a new disease-modifying antirheumatic drug (DMARD), among patients with early rheumatoid arthritis (RA).MethodsPatients newly diagnosed with...

Состояние костномозгового кроветворения у больных хронической лучевой болезнью, облученных с различной мощностью дозы

Purpose: to study the features of the state of the bone marrow according to cytological and histological studies at different periods of the course of CRD, caused by exposure to different dose rates, which developed as a result of professional prolonged exposure in a cohort of former employees of a radiation hazardous enterprise who underwent inpatient examination at the clinic of the A.I. Burnazyan Federal Medical and Biomedical Center in the period up to 1995. Material and methods: the study of the results of cytological and histological studies of the bone marrow was carried out in former workers of the Mayak Production Association who were exposed to long-term industrial exposure with a dose rate of 0.008−0.07 Gy / day (15 people), 0.003−0.007 Gy / day (12 people) and less than 0.001 Gy/day (25 people), during periods of formation, outcomes, immediate and long-term consequences of CRS. At the stage of studying the results of histological examination of the bone marrow, 54 more patients with CRS were added to the third group, irradiated with a dose rate of less than 0.001 Gy / day. Statistical processing of the material was carried out using the IBM SPSS Statistics software package.23 using the Kruskal‒Wallis test and the Mann‒Whitney U-test for independent samples. The results obtained were considered statistically significant at p &lt; 0.05. Results: At a dose rate of 0.008−0.07 Gy / day during the formation period, the myelogram revealed narrowing of the granulocyte and expansion of the red germs, acceleration of maturation of granulocytes with a normal erythrocyte maturation index and leuko-erythroblast ratio. In peripheral blood – agranulocytosis. In the period of outcomes and immediate consequences − narrowing of the granulocytic, expansion of erythrocyte germs, acceleration of maturation of neutrophils with other myelogram parameters within normal limits. In the blood − agranulocytosis, anemic syndrome. In the long term, in the case of restoration of hematopoietic function, narrowing of the granulocytic germ with other myelogram parameters within the reference values. However, with a similar course of CRS in 60 % of patients in periods of outcomes and long-term consequences, the development of myelodysplastic syndrome with transformation into acute leukemia or aplastic anemia is possible. At an irradiation dose rate of 0.003−0.007 Gy / day in the period of CRD formation, the myelogram revealed an expansion of the granulocytic germ, with other indicators within the normal range. In the period of outcomes and immediate consequences, an acceleration of maturation of granulocytes and an increase in the leuko-erythroblastic ratio were found. In the long term, a narrowing of the granulocytic and expansion of the erythrocyte sprouts, a slight acceleration of the maturation of granulocytes were found. Hystological examination: polymorphocellular bone marrow – in 11 out of 25 patients, hypoplasia – in 9 out of 25, signs of hyperplasia – in 5 out of 25. In the long term, 2 patients from this group developed oncohematological diseases. At an irradiation power of less than 0.001 Gy / day, during all periods of CRD, normal values of granulocytic and erythrocyte sprouts were noted in the myelograms of patients. In the period of CRD formation, the normal size of the granulocytic germ was achieved due to the accelerated maturation of neutrophils. The leuko-erythroblastic ratio in the period of long-term consequences was significantly higher than the norm (5.29 and 4.5, respectively). Hystological examination: 32 out of 64 patients had polymorphocellular bone marrow, 22 out of 64 had hypoplastic bone marrow, 7 out of 64 had bone marrow hyperplasia. Conclusion: regular changes in hematopoietic tissue and peripheral blood in CRD can serve as diagnostic criteria, based on which it is possible to assume the dose rate of radiation to which the patient was exposed, and also on their basis, it is possible to predict the outcome and long-term consequences of CRD that developed as a result of this exposure.

Diagnostic difficulties in polymyalgia rheumatica cases with normal erythrocyte sedimentation rate and C-reactive protein values.

Polymyalgia Rheumatica (PMR) is an inflammatory disease which does not have specific diagnostic tests or pathological symptoms and is identified based on clinical characteristics. Among acute phase reactants (APR), the erythrocyte sedimentation rate (ESR) and C-Reactive Protein (CRP) are laboratory findings used in diagnosis and follow-up. In this study, it was aimed to determine the incidence of normal ESH and CRP in patients diagnosed with PMR and identify the distinguishing characteristics of these patients. PMR patients who were clinically diagnosed at a single center were reviewed. After the presence of bursitis was demonstrated with ultrasonography in patients with normal ESR and CRP rates, they were accepted to have PMR. Among all 54 patients (63% female), ESR and CRP values were normal in 8 patients (14%), and serum amyloid A (SAA) was determined to be elevated in all these patients. In the comparisons of the groups with normal and high levels of ESR and CRP, it was found that the group with normal ESR and CRP values had a younger age of diagnosis (P = .027), a longer symptom duration (P < .001), and a lower comorbidity rate (P = .010). PMR patients can have normal ESR and CRP values at the time of their diagnosis. While bursitis can be demonstrated with ultrasonography in patients who are clinically evaluated to have PMR, APRs such as SAA other than ESR and CRP can also be used.

The Development of Magnolol-Loaded Intravenous Emulsion with Low Hepatotoxic Potential.

Intestinal failure-associated liver disease (IFALD) is a severe liver injury occurring due to factors related to intestinal failure and parenteral nutrition administration. Different approaches are studied to reduce the risk or ameliorate the course of IFALD, including providing omega-3 fatty acids instead of soybean oil-based lipid emulsion or administering active compounds that exert a hepatoprotective effect. This study aimed to develop, optimize, and characterize magnolol-loaded intravenous lipid emulsion for parenteral nutrition. The preformulation studies allowed for chosen oils mixture of the highest capacity of magnolol solubilization. Then, magnolol-loaded SMOFlipid was developed using the passive incorporation method. The Box-Behnken design and response surface methodology were used to optimize the entrapment efficiency. The optimal formulation was subjected to short-term stress tests, and its effect on normal human liver cells and erythrocytes was determined using the MTT and hemolysis tests, respectively. The optimized magnolol-loaded SMOFlipid was characterized by the mean droplet diameter of 327.6 ± 2.9 nm with a polydispersity index of 0.12 ± 0.02 and zeta potential of -32.8 ± 1.2 mV. The entrapment efficiency of magnolol was above 98%, and pH and osmolality were sufficient for intravenous administration. The magnolol-loaded SMOFlipid samples showed a significantly lower toxic effect than bare SMOFlipid in the same concentration on THLE-2 cells, and revealed an acceptable hemolytic effect of 8.3%. The developed formulation was characterized by satisfactory stability. The in vitro studies showed the reduced cytotoxic effect of MAG-SMOF applied in high concentrations compared to bare SMOFlipid and the non-hemolytic effect on human blood cells. The magnolol-loaded SMOFlipid is promising for further development of hepatoprotective lipid emulsion for parenteral nutrition.

Classification of Microscopic Malaria Parasitized Images Using Deep Learning Feature Fusion

An infectious disease that causes a chronic and potentially life-threatening infection caused by microorganisms of the Plasmodium class, is malaria, or malarial disease. It is critical to detect the presence of Malaria parasites as early as possible to ensure that antimalarial treatment is adequate to cure the particular type of Plasmodium. This is to reduce death rates and to focus on various infections in the event of an adverse outcome. The purpose of this study was to develop an artificial intelligence approach capable of separating parasitized erythrocytes from normal basophilic erythrocytes as well as platelets overlying the red blood cells to overcome the high cost of Ma-laria diagnostic equipment. The tone and texture characteristics of erythrocyte images were extracted using histo-gram thresholds and watershed methods, and then fused with Squeeze Net and ShuffleNet algorithms. The measures included planning, preparing, approving, and testing Deep Convolution Neural Network Segmentation without preparation using a graphic processor unit. A total of 96 percent accuracy and specificity was obtained for the position of malaria in red blood cells based on the results of all of the tests. It has been demonstrated that deep learning can be effective in the field of clinical pathology. This provides new directions for development as well as increasing awareness of researchers in this field.

Avian Erythrocytes and Agranulocytes - a Review

This study reports on the morphology of red blood cells and agranulocytes in various types of birds, such as rheas, emus, ostriches, chickens, and geese. Though all of them have the typical avian similarity in blood cells, yet there are some differences. Along with normal and mature erythrocytes, lymphocytes, and monocytes, the present study also focuses on rubricytes, reticulocytes, poikilocytes, polychromatophilic erythrocytes, reactive lymphocytes, pleomorphic monocyte nuclei, etc.

Mancozeb-induced cytotoxicity in human erythrocytes: enhanced generation of reactive species, hemoglobin oxidation, diminished antioxidant power, membrane damage and morphological changes

Mancozeb is an ethylene bis-dithiocarbamate fungicide extensively used in agriculture to safeguard crops from various fungal diseases. The general population is exposed to mancozeb through consumption of contaminated food or water. Here, we have investigated the effect of mancozeb on isolated human erythrocytes under in vitro conditions. Erythrocytes were treated with different concentrations of mancozeb (0, 5, 10, 25, 50, 100 μM) and incubated for 24 h at 37 °C. Analysis of biochemical parameters and cell morphology showed dose-dependent toxicity of mancozeb in human erythrocytes. Mancozeb treatment caused hemoglobin oxidation and heme degradation. Protein and lipid oxidation were enhanced, while a significant decrease was seen in reduced glutathione and total sulfhydryl content. A significant increase in the generation of reactive oxygen and nitrogen species was detected in mancozeb-treated erythrocytes. The antioxidant capacity and the activity of key antioxidant enzymes were greatly diminished, while crucial metabolic pathways were inhibited in erythrocytes. Damage to the erythrocyte membrane on mancozeb treatment was apparent from increased cell lysis and osmotic fragility, along with the impairment of the plasma membrane redox system. Mancozeb also caused morphological alterations and transformed the normal discoid-shaped erythrocytes into echinocytes and stomatocytes. Thus, mancozeb induces oxidative stress in human erythrocytes, impairs the antioxidant defense system, oxidizes cellular components, that will adversely affect erythrocyte structure and function.

Hubungan Kadar Feritin Serum dengan Tes Fungsi Hati pada Pasien Thalasemia Mayor

Thalassemia is a disease characterized by the destruction of red blood cells so that the life of red blood cells becomes shorter than normal erythrocytes. Thalassemia sufferers need routine blood transfusions throughout their lives, which results in a buildup or excess iron that causes damage to the body's organs, one of which is the liver. Iron in the body binds to ferritin and is stored in the liver. Ferritin examination is needed to see the excess or storage of iron in the body, while SGOT and SGPT are required to describe liver cell damage due to iron hoarding. This study aims to see the relationship between serum ferritin levels and liver function tests and the connection between adherence to consuming iron chelation and ferritin levels of thalassemia major patients. This study is an analytical observational study using a Cross-Sectional approach, with the subjects of the study being 22 children of Thalassemia Major who met the inclusion criteria for Ferritin, SGOT, and SGPT examinations. The results of the Spearman correlation test showed no relationship between serum ferritin levels with SGPT levels (sig. value 0.776) or SGOT (sig. value 0.766). Eta's correlation test for adherence to consuming iron chelation will affect serum ferritin by 36.2% in Thalassemia Major children at Dr. Tjitrowardojo Purworejo Hospital.