Decrease In Red Blood Cells Research Articles

Impact of zinc oxide nanoparticles on the behavior and stress indicators of African catfish (Clarias gariepinus) exposed to heat stress

This study was designed to assess the role of nano-zinc oxide in mitigating the deleterious effects of heat stress in African catfish (Clarias gariepinus) by evaluating parameters such as aggressive behavior (biting frequency and chasing duration), hematological indicators, and stress-related biochemical markers. A total of 96 catfish were divided into four distinct groups (24 fish/group): The first group (CON) served as the control group, receiving a diet free of nano-zinc oxide. The second group (HS) was exposed to heat stress at 35 °C ± 1 °C. The third group (ZN) was fed a diet containing nano-zinc oxide at 30 mg/kg of the diet, and the fourth group (ZHN) was exposed to heat stress (35 °C ± 1 °C) and fed a diet containing nano-zinc oxide at 30 mg/kg of the diet. The results clarified that the aggressive behavior and cortisol levels were significantly higher (P < 0.05) in the HS group compared to the CON and ZHN groups. Additionally, the level of acetylcholinesterase (AChE) was significantly lower (P < 0.05) in the HS group compared to the CON and ZHN groups. Meanwhile, a significant (P < 0.05) decrease in red blood cells, hemoglobin, packed cell volume, white blood cells, alkaline phosphatase, and lymphocytes, was observed in fish belonging to the HS group, while the levels of alanine aminotransferase, aspartate aminotransferase, neutrophils, and monocytes showed a significant increase (P < 0.05). Supplementation with nano-zinc oxide significantly recovered most hematological and biochemical parameters. In conclusion, nano-zinc oxide contributed significantly to the regulation of the negative impacts of heat stress on fish by reducing aggressive behavior and cortisol levels. Additionally, it improved the levels of AChE and certain hematological and biochemical parameters.

ANEMIA HEMOLÍTICA IMUNOMEDIADA SECUNDÁRIA A ERLIQUIOSE

Ehrlichiosis is an infectious and contagious disease that can be transmitted to humans and is therefore considered a zoonosis. During the development of the disease, immune- mediated haemolytic anemia occurs, characterized by a decrease in red blood cells due to the action of the phagocytic system in association with the presence of immunoglobulins. This research is based on the relationship between immune-mediated hemolytic anemia and Ehrlichiosis, with the aim of understanding both diseases, relating how they develop in the body, as well as their etiology, symptoms, laboratory changes, treatment and prophylaxis. To achieve this goal, data was collected through bibliographic research, and the results indicated the importance of controlling and preventing the disease through measures linked to the use of ticks and periodic monitoring by the veterinarian, since immune-mediated hemolytic anemia occurs secondary to this hemoparasitosis.

Distribution and risk factors of Trichomonas vaginalis infection with relevance to interleukin profile among population in Duhok city, Iraq

BackgroundVaginal trichomoniasis is a sexually transmitted disease causes adverse health outcomes, the causative agent is Tichomonas vaginalis. Infected women develop abnormal vaginal discharge with vaginitis. While, men mostly suffer from prostatitis and urethritis. ObjectivesThis study aimed to detect the rate of T.vaginalis infection in association with risk factors, measurement of blood parameters, interleukine-2 (IL-2) and interleukine-8(IL-8) levels among individuals. MethodsA total of 500 females were tested, a vaginal swab and blood were collected from the participants, and they were within the age range of 18–70 years old. High vaginal swabs were examined microscopically using wet mount preparation, the blood used for measurement of blood parameters using coulter count machine and IL-2 and IL-8 levels were evaluated using ELISA technique. ResultsOut of 500 individuals, the prevalence of T. vaginalis infection was 45(9.0 %). The maximum infection rates were reported among age group (18–28) years, rural residents, illiterates, married women (16.7 %), (15.9 %),(30.0 %),(11.7 %),respectively. Moreover, high positivity of infection revealed among low economical level 12.5 % compared to only 1.7 % in high economy group. Unemployed women, those with bad vaginal hygiene showed high incidence of infection 10.5 % and 6.3 %, respectively. It was noted a significant decrease in red blood cells and hemoglobin concentration, however, there was a significantly increased in white blood cells among infected subjects. Furthermore, the levels of both interleukin-2 and interleukin-8 were significantly higher among infected individuals compared to healthy ones. ConclusionsEffective strategies for disease awareness are needed and should include health promotion, education, and prevention.

Traditional Chinese medicine formulation ChangQing compound has significant therapeutic effects on chickens infected with Eimeria tenella

Coccidiosis poses a significant challenge to the poultry industry. However, the excessive and improper use of anticoccidial drugs and vaccines has led to resistance and food safety concerns. Consequently, traditional Chinese herbs have garnered attention as a potentially safer and more effective alternative. ChangQing compound derived from various Chinese herbal medicines is a promising anticoccidiosis agent, but its therapeutic effects have not been comprehensively evaluated. This study aimed to assess the therapeutic efficacy of ChangQing Compound against Eimeria tenella-induced coccidiosis in chickens on the basis of physiological indicators, cecum lesions, and changes in microbial diversity. The comparison with the positive control group revealed the average weight gain (AWG) and anticoccidial index (ACI) of the chicks were significantly higher, in contrast, the feed conversion ratio (FCR), cecal lesion score (CLS), and oocyst count per gram of cecal content (OPG) were significantly lower (P<0.05). Notably, AWG (138.87 g), OPG (0.57 × 106), ACI (177.92), and FCR (2.51) reflected the significant therapeutic effect of the 2.5 g/L ChangQing compound treatment (CQM). Histological sections showed that the cecal villus damage and intestinal wall swelling were minimal in the CQM, consistent with the CLS (0.73). Additionally, the 2.5 g/L ChangQing compound treatment effectively prevented the decrease of red blood cells, platelets, and hemoglobin, while promoting the release of anti-inflammatory factors interleukin-10 and interleukin-4, and inhibiting the pro-inflammatory factors interferon-γ and interleukin-17. The microbial community structure in the CQM was most similar to that of the negative control group. In summary, ChangQing compound had multiple positive effects (e.g., promoting weight gain, alleviating anemia, suppressing coccidial proliferation, reducing intestinal damage, modulating immunity, and maintaining intestinal microbiota homeostasis). The study results may be relevant to developing a novel strategy for the clinical management of coccidiosis.

Safety and efficacy of olutasidenib treatment in elderly patients with relapsed/refractory mIDH1 acute myeloid leukemia.

6527 Background: Elderly patients with relapsed/refractory (R/R AML) are a special population that may elevate safety concerns, which must be balanced with clinical benefit. Olutasidenib (OLU), a targeted therapy for m IDH1 AML, was well tolerated in a registrational, Ph2, open-label, multicenter trial and, in the pivotal cohort, demonstrated complete remission (CR) or CR with partial hematologic recovery (CRh) in 35% of patients (age 32-87 years, y) for a median duration of 25.9 months. We evaluated safety and efficacy of OLU in the elderly subgroup (≥75y) with m IDH1 AML. Methods: In the Phase 2 trial, the pivotal cohort enrolled adults with R/R m IDH1AML. OLU was administered at 150 mg BID. Adverse events (AE) and lab values were collected at least monthly and graded based on the NCI CTCAE Version 4.03. Efficacy was based on response criteria of the International Working Group in AML (2003). Results: 153 patients received OLU monotherapy, 147 had centrally-confirmed mIDH1 AML, , and 45 were ≥75y (range 75-87) at study entry. Of the 45 elderly patients, 27 (60%) were male, 51% had ECOG=1, 16% had ECOG =2, 71% were relapsed, 29% had refractory AML, 58% had prior HMA, 11% received prior venetoclax, and none had prior stem cell transplant. Median bone marrow blasts were 43% (5-93). Genetic co-mutations included FLT3 in 9%, NPM1 in 20%, and TP53 in 2%. Grade 3 or 4 adverse events were reported in 45.8% of elderly patients; the most common were decreases in red blood cells (31%), platelets (13%) and neutrophils (10%) as well as febrile neutropenia (15%). AEs were consistent with published pivotal Phase 2 results, with no new safety signals. Deaths occurred in 19 patients due to disease progression (9), pneumonia (2), sepsis (2), GI haemorrhage (1), aortic stenosis (1), atrioventricular block (1), cardiac failure congestive (1), acute kidney injury (1) and other (1). The response to OLU therapy is summarized in the table. 31% of patients ≥75 y achieved CR/CRh; median time to CR/CRh was 1.5 mos (0.9-5.6) and median duration of CR/CRh was 25.9 mos (95%CI 7.4, not reached). Conclusions: Olutasidenib was generally well tolerated in elderly patients with R/R m IDH1AML and induced durable remissions. Despite the challenges of treating elderly patients who had already failed prior AML treatment, the results suggest that elderly patients can benefit from therapy with olutasidenib. Clinical trial information: NCT02719574 . [Table: see text]

Kegiatan Skrining Pemeriksaan Hemoglobin dan Hematokrit terhadap Penyakit Anemia pada Kelompok Lanjut Usia di Panti Werda Hana

Anemia in the elderly is a common condition characterized by a decrease in red blood cells or hemoglobin levels, causing various health problems. Anemia is defined as a hemoglobin level below 13 g/dL in men and 12 g/dL in women. Various factors, such as inadequate nutritional intake, chronic inflammation, and kidney disease, often cause anemia in this population. Untreated anemia impacts quality of life, increases hospitalization rates, the risk of falls, and cognitive decline, and is independently associated with higher mortality from cardiovascular stress. This activity uses the PDCA (Plan-Do-Check-Action) method to screen and treat anemia in elderly participants Hana Nursing Home. We conducted complete blood tests on 61 respondents to evaluate their hematological profile. The blood test results revealed an average hematocrit of 36.15% for the participants, and anemia in 11 (18.03%) men and 19 (31.15%) women. Routine hemoglobin screening and effective management are essential for early detection and intervention, thereby improving the quality of life of the elderly. Keywords: Anemia, Elderly, Hemoglobin, Hematocrit

Characterization and clinical significance of hemolysis after pulsed-field ablation for atrial fibrillation: insights from a multicentric analysis

Abstract Background Pulsed-field ablation is a novel non-thermal ablation modality which is being increasingly used in clinical practice for the treatment of atrial fibrillation (AF). While the susceptibility of erythrocytes to electroporation is well-established, the effect of cardiac PFA technologies on erythrocyte destruction has not been described so far. Purpose The aim of this study was to investigate whether PFA induces clinically significant hemolysis. Methods We included 187 patients undergoing AF catheter ablation with either PFA (n=145) or RFA (n=42) at four high-volume centers. PFA was performed using a pentaspline catheter (biphasic, bipolar pulses of 2kV) and RFA with a 3.5 mm irrigated-tip catheter (40-60 W). The lesion set comprised pulmonary vein isolation (PVI) for paroxysmal AF and PVI ± additional lesions/lines for persistent AF. Established biomarkers of hemolysis, anemia and renal function were analyzed in blood samples obtained at baseline (T1), at the end of ablation (T2) and 24 hours after the procedure (T3). Results Baseline characteristics were well-balanced between groups (65.5 ± 9.6 years, 72.7% male, p&amp;gt;0.05). The ablation procedures comprised a mean of 61.4 ± 27.2 deliveries (PFA group) and a mean RF duration of 31.0 ± 16.3 min (RFA group). Atrial ablation in addition to PVI was performed in 55.2% vs. 61.9% patients, respectively (p=0.469). The direct hemolysis marker free plasma hemoglobin was significantly higher with PFA (592.8 ± 330.6 mg/dl) than with RFA (186.6 ± 215.8 mg/dl, p&amp;lt;0.001) at the end of ablation. A strong correlation was found between free plasma hemoglobin and PFA deliveries (Pearson r=0.615, p&amp;lt;0.001). PFA was further associated with a 2.2-fold increase in bilirubin (21.3 ± 11.3 vs. 9.6 ± 4.7 µmol/l), a 2.4-fold decrease in haptoglobin (0.5 ± 0.4 vs. 1.2 ± 0.4 g/l) and a 1.8-fold increase in LDH (352.7 ± 115.7 vs. 192.2 ± 51.6 U/l, T3 vs. T1, all p&amp;lt;0.001). There were no significant changes in bilirubin and haptoglobin in the RFA group (p&amp;gt;0.05). A slight decrease in red blood cells, hemoglobin and hematocrit was detected after both PFA and RFA (Abstract Figure), with no significant differences between groups (all p&amp;gt;0.05). While mean creatinine levels and glomerular filtration rates did not significantly change after ablation in either group, acute kidney injury occurred in 4/124 (3.2%) with PFA vs. 0/32 (0%) with RFA (p=0.303). Conclusion Intravascular hemolysis is a frequent finding after PFA for AF and increases with the number of PFA deliveries. While PFA was not associated with clinically significant anemia, the incidence of acute kidney injury in the present study warrants future large-scale investigations assessing the biological impact and nephrotoxicity of PFA-induced hemolysis.Abstract Figure

Mentha piperita and Thymus vulgaris ameliorate hematological changes induced by chronic consumption of oxidized palm oil in rats

Objective: The aim of this study was to investigate the effect of Mentha piperita L and/or Thymus vulgaris in ameliorating the hematological changes induced by chronic consumption of oxidized palm oil (OPO) in rats. Methodology: Fifty rats were divided into five groups as follows: rats received a basal diet (control), rats received a basal diet sustained with OPO, rats fed OPO and received Mentha piperita extract, rats fed OPO and received Thymus vulgaris extract, and finally rats fed OPO and received both extracts. After 6 weeks, blood samples were withdrawn and a complete blood count test was done. Results: Rats fed a diet containing OPO showed a significant decrease in red blood cells (RBCs), hematocrit (Hct), mean corpuscular volume (MCV), hemoglobin (Hb), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), lymphocytes (LYM), and platelets (PLTs) accompanied by a significant increase in white blood cells (WBCs), red cell distribution width (RDW), and mean platelet volume (MPV) compared to rats fed a normal diet. In contrast, rats fed on diet containing OPO and the aqueous extract of either Mentha piperita or Thymus vulgaris showed a significant increase in RBCs, Hct, MCV, Hb, MCH, MCHC, LYM, and PLTs accompanied by a significant decrease in RDW, WBCs, and MPV relative to rats fed on diet containing OPO only. Treatment with a combination of both plants showed better improvements. Conclusion: Both Mentha piperita and Thymus vulgaris ameliorate the hematological changes induced by OPO.

Early differential diagnosis of pancytopenia related diseases based on serum surface-enhanced Raman spectroscopy

Pancytopenia is a common blood disorder defined as the decrease of red blood cells, white blood cells and platelets in the peripheral blood. Its genesis mechanism is typically complex and a variety of diseases have been found to be capable of causing pancytopenia, some of which are featured by their high mortality rates. Early judgement on the cause of pancytopenia can benefit timely and appropriate treatment to improve patient survival significantly. In this study, a serum surface-enhanced Raman spectroscopy (SERS) method was explored for the early differential diagnosis of three pancytopenia related diseases, i.e., aplastic anemia (AA), myelodysplastic syndrome (MDS) and spontaneous remission of pancytopenia (SRP), in which the patients with those pancytopenia related diseases at initial stage exhibited same pancytopenia symptom but cannot be conclusively diagnosed through conventional clinical examinations. The SERS spectral analysis results suggested that certain amino acids, protein substances and nucleic acids are expected to be potential biomarkers for their early differential diagnosis. In addition, a diagnostic model was established based on the joint use of partial least squares analysis and linear discriminant analysis (PLS-LDA), and an overall accuracy of 86.67 % was achieved to differentiate those pancytopenia related diseases, even at the time that confirmed diagnosis cannot be made by routine clinical examinations. Therefore, the proposed method has demonstrated great potential for the early differential diagnosis of pancytopenia related diseases, thus it has significant clinical importance for the timely and rational guidance on subsequent treatment to improve patient survival.

Insight into hematological parameters of petrol station workers.

Benzene is one of the major carcinogenic factors that can affect liver, kidneys, and lungs. Chronic inhalation of benzene vapor by petrol stations workers has been shown to have an impact on hematological parameters; thus, the present study aimed to investigate the effect of benzene exposure on petrol station workers. The study involved 99 participants, 50 of whom have been exposed to benzene and 49 of whom have not (control). A 5 ml blood sample in an ethylenediaminetetraacetic acid (EDTA) anticoagulant tube was collected from each subject, and a complete blood count test was used to test hematological parameters. The current study showed a significant decrease in red blood cells, packed cell volume, and hemoglobin in the exposed group compared to the control group. However, the amount of white blood cells was significantly increased (p < 0.0001) in the exposed group compared to the control group. Notably, there was no significant difference in platelet counts between the two groups. In terms of exposure time, subjects who have been exposed to benzene for more than a year and fewer than 10 years showed a significant decrease (p < 0.05) in RBCs indices and a significant increase (p < 0.0001) in WBCs compared to those in the control group CONCLUSIONS: Thus, the findings indicated that significant differences in hematological parameters were found in workers who were exposed to benzene compared to those who had not been exposed.

Doppler ultrasonographic scan, gene expression and serum profile of immune, APPs and antioxidant markers in Egyptian buffalo–cows with clinical endometritis

The objective of this study was to elaborate Doppler ultrasonographic scan, genetic resistance and serum profile of markers associated with endometritis susceptibility in Egyptian buffalo–cows. The enrolled animals were designed as; twenty five apparently healthy buffalo–cows considered as a control group and twenty five infected buffalo with endometritis. There were significant (p < 0.05) increased of cervical diameter, endometrium thickness, uterine horn diameter, TAMEAN, TAMAX and blood flow through middle uterine artery with significant decrease of PI and RI values in endometritis buffalo–cows. Gene expression levels were considerably higher in endometritis-affected buffaloes than in resistant ones for the genes A2M, ADAMTS20, KCNT2, MAP3K4, MAPK14, FKBP5, FCAMR, TLR2, IRAK3, CCl2, EPHA4, and iNOS. The RXFP1, NDUFS5, TGF-β, SOD3, CAT, and GPX genes were expressed at substantially lower levels in endometritis-affected buffaloes. The PCR-DNA sequence verdicts of healthy and affected buffaloes revealed differences in the SNPs in the amplified DNA bases related to endometritis for the investigated genes. However, MAP3K4 elicited a monomorphic pattern. There was a significant decrease of red blood cells (RBCs) count, Hb and packed cell volume (PCV) with neutrophilia, lymphocytosis and monocytosis in endometritis group compared with healthy ones. The serum levels of Hp, SAA, Cp, IL-6, IL-10, TNF-α, NO and MDA were significantly (P˂0.05) increased, along with reduction of CAT, GPx, SOD and TAC in buffalo–cows with endometritis compared to healthy ones. The variability of Doppler ultrasonographic scan and studied genes alongside alterations in the serum profile of investigated markers could be a reference guide for limiting buffalo endometritis through selective breeding of natural resistant animals.

Influence of Frequent Phlebothomy on Blood Iron Concentration, Haematological, Metabolic and Endocrine Parameters in Rams

Abstract Frequent phlebotomies, even when only a small volume of blood is taken for diagnostic purposes, can lead to the development of iron deficiency with hematological and metabolic changes. The study aimed to determine the influence of frequent blood loss by phlebotomy on blood iron concentration (Fe), hematology, metabolic and endocrine parameters and their relationships. Blood samples were collected from 30 blood donor rams for 6 consecutive weeks, with approximately 10% of blood collected weekly. Such chronic blood loss resulted in a decrease in Fe. Indicators of iron transport in the bloodstream changed, so the value of total iron-binding capacity (TIBC) and unsaturated iron-binding capacity (UIBC) increased, while the value of transferrin saturation percentage (TS%) decreased. Hematological changes included a decrease in red blood cells, hemoglobin, mean red blood cell volume and hematocrit and a tendency for reticulocyte count and red cell distribution width to increase. Chronic blood loss resulted in a specific metabolic response that included the increase in glucose, cholesterol, triglycerides, aspartate-aminotransferase, and insulin resistance, while thyroxine, triiodothyronine and cortisol decreased and there was a tendency for lactate to increase and BHB to decrease. The mentioned blood parameters correlated with Fe and additionally showed greater changes when Fe was extrapolated to the level of clinical deficit (Fe=9μmol/L). These correlations suggest the need to monitor the metabolic and endocrine status during chronic blood loss, in addition to Fe and erythrocyte indices. Compared with previous results in other animal species, Fe may have a direct influence on metabolic processes in rams.

THEILERIA EQUI INFECTION IN WORKING HORSES OF PAKISTAN: EPIDEMIOLOGY, MOLECULAR CHARACTERIZATION, AND HEMATOBIOCHEMICAL ANALYSIS.

Theileria equi is 1 of the emerging and prevailing tick-borne hemoprotozoans adversely affecting the equids worldwide, including Pakistan. The current study aimed to investigate the prevalence and molecular characterization of T. equi in working horses (n = 194), the comparative efficacy of different diagnostic tests, associated risk factors, and hematobiochemical analysis. The blood samples of horses were subjected to microscopic examination, cELISA, and polymerase chain reaction (PCR) and the results revealed a prevalence of 9.79, 21.13, and 13.40%, respectively, for T. equi in working horses. The comparison of microscopy and cELISA results with PCR showed that cELISA had higher sensitivity (84.62%), but lower specificity (88.69%) and accuracy (88.14%) in comparison to microscopy (57.69, 97.62, and 92.27%). Molecular characterization of T. equi by phylogenetic analysis revealed a 61% resemblance of study isolates with each other OL662926, OL662925, and 82% similarity with isolate OL662924 while also showing homology with T. equi isolates of South Africa, South Korea, India, Pakistan, and Brazil. The risk factor analysis revealed a significant association (P < 0.05) of tick control status, previous tick history, tick infestation, house hygiene, deworming/vaccination, and the presence of other livestock species with T. equi infection in horses. The hematobiochemical profile revealed a significant (P < 0.05) decrease in red blood cells (RBCs), hemoglobin (Hb), packed cell volume (PCV), white blood cells (WBCs), platelet (PLT), phosphorus, and an increase in lymphocytes, granulocytes, aspartate aminotransferase (AST), glucose, bilirubin, blood urea nitrogen (BUN), and creatinine in T. equi-infected horses. The current study is the first comprehensive report for comparative evaluation of microscopy, cELISA, and PCR, assessment of epidemiological risk factors as well as hematobiochemical variations due to T. equi infection in Pakistan.

Effect of Some natural products against Ehrlich Ascites Carcinoma in mice: A Review

Cancer is a serious disease and it is one of the main causes of death, and it is considered the second cause, after people who suffer from heart failure. Through the statistics, Breast cancer is one of the most common types of cancer, with a rate of 99% in women and 1% in men. It is very complex, and patients with this type of cancer may face numerous challenges during their treatment period. It was determined that the death rate from cancer had decreased by 1.5%, but it is expected to increase in the future due to an increase in cancer cases (breast, prostate). Ehrlich model is a cancer that grows inside a mouse. Its origin is spontaneous mammary carcinoma, and when it spreads within the mouse, it takes two forms, either ascitic or solid. Ehrlich's ascites carcinoma is a spontaneous breast cancer that has a very high importance in scientific research. It is undifferentiated and has a short life. This type of cancer was used for the first time in 1932, when it was used inside the mouse in the abdomen, and a type of fluid was obtained for this called ( Ehrlich's ascites) due to the liquid substance on which cancer cells depend and consider it food and a means for rapid tumor growth, EAC deficiency causes anemia and thus causes a decrease in red blood cells or hemoglobin, EAC causes an increase in urea and a decrease in creatinine, and EAC causes an increase in liver enzymes and a decrease in total protein.Natural products are the only and most effective products that lead to the development of drugs, especially anti-cancer drugs and anti-inflammatories. Through studies, it has become clear that natural products are a source of drugs.

Assessment of Ferritin Level and Some Biochemical Variables in Iraqi Anemic Diabetic Mellitus in Both Gender

Diabetes mellitus (DM) is a prevalent metabolic disorder characterized by elevated blood sugar levels caused by diminished tissue sensitivity to insulin, insufficient insulin production, or a combination of both factors. This is a non-communicable disease that is also highly prevalent globally. The aim of the study is to evaluate ferritin level and other biochemical variables in type 2 diabetes mellitus patients and study the effect of biochemical variables between male and female (T2DM) . Method involves enrollment of 120 participant divided into two main groups (T2DM and control group) and every group has been subdivided based on gender to (30 male and 30 female) for each group. All the groups studied for determination of anthropometric and biochemical parameters to evaluate the relationship between the studied parameters and T2DM patients. Results: the result showed that (Age, body mass index (BMI), Fasting blood sugar (FBS), triglycerides (TC), Urea, systolic blood pressure (SBP), diastolic blood pressure (DBP) and Ferritin) were significantly difference in T2DM patients (male and female) as compared with control. While ( TG and Creatinine) were non-significantly differ in T2DM patients as compared with control. Furthermore, there was non-significant difference in the studied parameters between T2DM males and females. Conclusion: from the obtained result it could be concluded that patients with T2DM have a problem associated with ferritin level that effected by T2DM and decreased because of the impaired renal dysfunction and red blood cells decrease and that causes a decrease in the level of ferritin. Age and BMI levels have a direct effect and correlated with elevation of lipid continents and high level of FBS and blood pressure. Renal function is highly elevated because of the T2DM complication, and it is conceder as a risk factor for T2DM.

Protective effects of Helicteres isora L. fruit extract on phenylhydrazine-induced hemolytic anemia in rats

BackgroundAnemia is a serious global public health problem that affects young children and pregnant women with an increased risk of maternal and child mortality. Helicteres isora L. (Family: Malvaceae) has significant antioxidant properties responsible for its medicinal properties. The present study was conducted to investigate the protective effects of aqueous fruit extract of Helicteres isora L. (AEHI) on phenylhydrazine-induced hemolytic anemia in rats. MethodsThe fruits of Helicteres isora L. (H. isora L.) were extracted with distilled water to obtain the AEHI. The AEHI was subjected to preliminary phytochemical analysis, iron estimation and in-vitro antioxidant assay. The HPTLC fingerprinting followed by HR-LCMS analysis of the isolated HPTLC fraction of AEHI was carried out to investigate the phytoconstitutents. Male Wistar albino rats were administered phenylhydrazine (40 mg/kg, i.p.) for two consecutive days to induce anemia. The effects of AEHI were investigated at three oral doses viz. 250, 500 and 750 mg/kg once daily for 13 days by estimating blood parameters and pathological changes in the liver, heart, spleen, kidney, lungs and bone marrow. ResultsThe phytochemical analysis showed the presence of alkaloids, tannins, phenolic compounds, carbohydrates and steroids in the AEHI. The iron content of AEHI was found to be 208 µg/ml. In-vitro antioxidant assays, the IC50 value of AEHI was found to be 90.01 µg/ml. Phenylhydrazine administration caused a significant decrease in red blood cells, hemoglobin, and hematocrit levels. These decreased levels were significantly improved by AEHI in a dose-dependent manner. Pathological changes in the liver, heart, spleen, kidney, lungs and bone marrow were prevented by AEHI treatment. ConclusionIt is concluded that the AEHI provides significant protection against phenylhydrazine-induced hemolytic anemia.

PRECLIINICAL SUB-ACUTE ORAL TOXICITY STUDY FOR ANTI-INFLAMMATORY EVALUATION OF METHANOL EXTRACT OF CUCURBITA PEPO L. SEEDS

: Cucurbita pepo, a plant known for its traditional use in managing various ailments, contains valuable phytoconstituents and nutritional components. Aim: This study aimed to assess the sub-acute toxicity of the methanol extract of C. pepo (MECP) seeds according to OECD 407 guidelines. Rats were orally administered MECP at 250, 500, and 1000 mg/kg doses for twenty-eight days. Throughout the study period, the animals were closely monitored for any signs of toxicity, and at the end, blood samples were collected for biochemical and hematological analyses. Histopathological examination was performed on vital organs after euthanizing the rats. The results demonstrated that MECP administration significantly increased body and organ weight in the treated rats. However, food and water intake remained unchanged, and no mortality was observed. Biochemical and hematological analyses revealed no significant changes in the treated groups compared to the control group, except for a notable decrease in red blood cells, suggesting potential anemic effects of MECP. Histopathological examination showed no structural alterations in the liver, heart, and kidney. Based on these findings, it can be concluded that C. pepo is safe for use and holds the potential for therapeutic efficacy.

Abstract B128: Preclinical characterization of ETx-22, a next-generation antibody drug conjugate (ADC) targeting nectin-4

Abstract Nectin-4 is overexpressed in a variety of cancers including urothelial, breast, cervix, lung and ovarian, with low level of expression in normal tissues. Enfortumab vedotin (EV), an anti-Nectin-4 ADC with a monomethyl auristatin (MMAE) payload, has been a significant advancement in the treatment of urothelial cancer. Yet, it is associated with important skin toxicity owing to the expression of nectin-4 in the skin, and peripheral neuropathy secondary to its toxin. We have developed a next-generation anti-nectin-4 ADC, ETx-22, which consists of a humanized IgG1, Fcg-silent antibody that is bound via maleimide-ß-glucuronide poly-sarcosine linkers to 8 molecules of the topoisomerase-I inhibitor, exatecan. Here, we describe the key pharmacological experiments, in-vivo efficacy and results from the good laboratory practice (GLP) toxicology studies. In-vitro studies have shown that the ETx-22 antibody exhibits a high affinity for tumor-expressed Nectin-4, but 10-fold lower affinity for nectin-4 expressed on skin, with significantly higher internalization in tumor cells. In-vivo efficacy was evaluated in various patient-derived xenograft models. ETx-22 was shown to be consistently superior to EV as well as chemotherapy, in a broad array of tumors, including urothelial, ovarian, cervical and triple negative breast cancer irrespective of the degree of nectin-4 expression. Antitumor activity was also observed in MMAE-resistant models. GLP studies evaluated repeated doses of ETx-22 in cynomolgus monkeys at two doses, 10 and 20 mg/kg. The treatment was administered as 30-minute intravenous infusions once every 2 weeks for a total of three cycles. The ETx-22 antibody alone was also evaluated at 20 mg/kg. Both ETx-22 doses were well tolerated and all animals survived up to scheduled euthanasia. The common observation at both doses was mild multifocal skin hyperpigmentation, which was reversible. No skin rash or pruritus was reported. Mild transient decrease in red blood cells was observed in between cycles. No gastrointestinal toxicities were observed. No signs of interstitial lung disease or eye toxicity were detected. Based on these findings, the highly non- severely toxic dose (HNSTD) is at least 20 mg/kg. The half-life of ETx-22 ranged from 3.36 – 3.6 days. In conclusion, preclinical and pharmacological shows that ETx-22 has better tolerance, longer half-life, and higher efficacy to EV. No skin rash was reported in toxicology studies. A first in human study (EXCEED) is planned to start in 2023. Citation Format: Hatem Azim, Florence Lhospice, Xavier Preville, Joanna Fares, Marc Lopez, Emerence Crompot, Emmanuelle Josselin, Remy Castellano, Maria Wehbeh, Yves Collette, Jack Elands, Daniel Olive. Preclinical characterization of ETx-22, a next-generation antibody drug conjugate (ADC) targeting nectin-4 [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr B128.

GDF11/SMAD Regulated Splicing of GATA1 Is Associated with Response to Luspatercept in Lower-Risk Myelodysplastic Syndromes (LR-MDS)

Introduction: Luspatercept is an erythroid maturation agent that promotes late-stage erythropoiesis in patients with LR-MDS. While it is known that luspatercept, a modified activin receptor ligand trap, binds to GDF11 and Activin B, the mechanism by which these ligands trigger a decrease in red blood cell (RBC) formation is unknown. Luspatercept was approved by the FDA following the phase 3 MEDALIST study, but biomarkers of response are currently unknown. Results:In a large cohort of primary MDS CD34+ samples, GDF11 expression was significantly higher in MDS samples than in age-matched healthy controls (183 MDS samples vs 17 controls, P &amp;lt; 0.05). GDF11 was detected in primary MDS sera and was substantially elevated in the refractory anemia (RA) and RA with ring sideroblasts (RARS) MDS subgroups. Activin receptor 2B (ACVR2B), the receptor of GDF11, was also overexpressed in MDS samples, and its expression was confirmed by immunohistochemistry (IHC) in the bone marrows of primary MDS patients. GDF11/ACVR2B activates SMAD2, and we found that the overexpression of SMAD2 in CD34+ cells was associated with more severe anemia in MDS patients. RNA-seq and SMAD2 ChIP-seq analyses were then performed to determine the downstream effects of GDF11 in erythroid progenitors. Human CD34+ primary cells were utilized to generate BFU-E stage erythroid progenitors. We discovered that GDF11 promotes an abnormal erythroid gene expression program characterized by increased expression of pro-apoptotic and early erythroid differentiation-related genes. SMAD2 binding was detected in regulatory regions of early erythroid (including GATA1), proinflammatory, and apoptotic transcripts using ChIP-seq analysis with p-SMAD2. In a zebrafish model, administration of GDF11 caused a decrease in erythroid cells, which was rescued by treatment with luspatercept (Wobus et al. Leukemia 2021 35(10):2936-47). Luspatercept inhibited the phosphorylation and activation of SMAD2 in response to GDF11 stimulation in primary hematopoietic cells and leukemia cell lines. Importantly, luspatercept could reverse the growth-inhibiting effects of MDS-derived sera on primary hematopoietic progenitors. Finally, to identify biomarkers associated with clinical responses, we analyzed phase 3 MEDALIST study samples collected before and after treatment with lupatercept. RNA-seq analysis was performed on bone marrow samples from responders (R; n = 14) and non-responders (NR; n = 9) (Figure). Analysis of pre-treatment/screening bone marrow transcriptomes revealed no significant differences between R and NR in global gene expression. Analysis of splice isoforms between the two cohorts revealed strikingly greater differences. Most differential splicing events were observed in NR (702 in R vs 5232 in NR). The most prevalent splicing modifications were exon skipping events, and most functional pathways affected by differential exon usage were RNA splicing related. GATA1, the master erythroid transcription factor, was spliced differently in R at baseline, which was an intriguing observation. Response to luspatercept was found to be associated with exon 2 skipping, resulting in a shortened isoform of GATA1. R had a greater proportion of the short GATA1 isoform than NR (12% in R versus 6% in NR; P = 0.04). We analyzed ChIP-seq data from primary erythroid cells to determine the effect of GDF11/SMAD2 activation on GATA1 splicing. GDF11 stimulation resulted in SMAD2 binding to the second intron of GATA1. CRISPR-mediated deletion of the SMAD2 binding region in primary erythroid progenitors increased expression of the GATA1 long isoform. Next, we performed a CRISPR-mediated deletion of exon 2 of GATA1, resulting in a predominance of GATA1 short isoform expression and a decrease in erythroid output. In CRISPR-modified primary erythroid cells with a predominant short GATA1 isoform (exon 2-deleted cells), luspatercept was able to reverse the inhibition induced by GDF11. Conclusions: These results demonstrate that MDS cases that respond to luspatercept are associated with a relatively high level of expression of functionally deficient GATA1 short isoform. We further demonstrate that GDF11 stimulates the production of the GATA1 short isoform via SMAD2 binding to intron 2 of the GATA1 gene. The regulation of a GATA1 splicing isoform by GDF11/SMAD2 is associated with clinical responses to luspatercept, according to these novel findings.

Rker-050, a Modified Activin Receptor Type Iia Ligand Trap, Promoted Erythropoiesis in a Murine Model of Myelofibrosis

Myelofibrosis (MF) is one of several myeloproliferative neoplasms most often associated with anemia due to ineffective hematopoiesis, inflammation, aberrant bone metabolism and progressive fibrosis in the bone marrow (BM). As a compensatory mechanism, extramedullary hematopoiesis is initiated in the spleen, resulting in splenomegaly. Megakaryocyte hyperplasia/dysplasia are thought to be one of the main drivers of MF disease progression. Evidence also suggests that dysregulated TGF-β superfamily activity contributes to ineffective hematopoiesis. KER-050 and its research form, RKER-050, are investigational modified activin receptor type IIA ligand traps designed to inhibit specific TGF-β superfamily ligands, including activin A, activin B, growth and differentiation factor (GDF) 8 and GDF11, to promote erythropoiesis and thrombopoiesis. Additionally, the KER-050 target ligands promoted fibrosis and bone resorption in preclinical studies. Therefore, by inhibiting these ligands, KER-050 could potentially rebalance the bone marrow microenvironment, including reducing fibrosis, allowing restoration of hematopoiesis and alleviating extramedullary hematopoiesis and associated splenomegaly. To investigate the potential of RKER-050 to reverse ineffective hematopoiesis in MF, a hypomorphic Gata1 MF mouse model (Gata1Low; MF mice) was utilized which presents similar characteristics to human MF, including ineffective hematopoiesis, cytopenias, extramedullary hematopoiesis, defective bone deposition and BM fibrosis. this study, we also observed aberrant bone growth in the cortex of the hind limb long bones which inappropriately extended into the bone marrow, resulting in limited BM compartment space and demonstrating a severe disease state in these mice. Anemia status of MF mice was confirmed at the start of the treatment. MF mice showed a 16% reduction in hemoglobin (Hgb) compared to WT control mice. After 12 weeks of treatment, MF+VEH mice continued to exhibit a significant or trending decrease in red blood cells (RBCs; -19%), Hgb (-20%), and hematocrit (Hct; -10%) compared to WT mice. In contrast, MF+050 mice had a significant recovery of RBCs (+31%), Hgb (+24%) and Hct (+20%), compared to MF+VEH mice, demonstrating that RKER-050 reversed anemia in this advanced disease MF model. To assess the mechanism of action of RKER-050, we evaluated erythroid precursor populations in the BM. While MF+VEH mice showed a significant reduction in erythroid progenitors compared to WT mice, These data support increased BM erythropoiesis with RKER-050 treatment. In addition, we evaluated the effect of RKER-050 on thrombopoiesis. MF+VEH mice were severely thrombocytopenic and had significantly increased megakaryocyte progenitors (MkP; +591%) compared to WT mice. Although RKER-050 significantly reduced MkP (-33%; MF+050), suggesting RKER-050 may positively influence the megakaryocyte lineage, this did not translate to improvements in thrombocytopenia, and platelet counts remained similar to MF+VEH. Additional data will be presented that investigates the potential of RKER-050 treatment in (6-month vs 10-month-old mice, where disease is less severe) to further evaluate the potential to impact cytopenias, extramedullary hematopoiesis and BM fibrosis. Overall, in the context of severe disease, our results suggest RKER-050 can promote erythropoiesis and reverse anemia in the BM in by rebalancing hematopoiesis in the BM niche. Reestablishing BM hematopoiesis could obviate the need for compensatory extramedullary hematopoiesis in the spleen, the major driver of splenomegaly in MF patients. In conclusion, KER-050 represents a potentially promising approach for patients with MF and other hematological diseases where ineffective hematopoiesis occurs.