Blood Cell Proliferation Research Articles

LITERATUR REVIEW : KUALITAS HIDUP PASIEN LEUKEMIA

Leukemia is a malignant disease that attacks the hematopoiesis system, causing uncontrolled blood cell proliferation. Quality of life is one indicator that can be used to see the welfare conditions of patients suffering from chronic diseases such as leukemia. This review literature aims to find out the factors that influence the quality of life of leukemia patients. Literature search through Google Scholar, EBSCO, Springer Link, Elsevier, Sage databases using keywords: factors, quality of life, leukemia. The journals taken have a range from 2018 to 2020, which are then filtered according to the inclusion criteria set by the author. The results of the review indicate that there are several factors that affect the quality of life in leukemia patients, namely the therapeutic treatment undertaken, the patient's environment, and the role of care giver. Proper management of leukemia patients will improve the quality of life of these patients, and vice versa.

Evaluation of the Antibacterial Activity, Acute Toxicity and Immuno-stimulatory Potential of Adenodolichos paniculatus root Extract

Medicinal plants have been used in the treatment of illnesses from time immemorial. In Nigeria today a large population of people still depend on herbal medicines for their health needs. The medicinal plant, Adenodolichos paniculatus has been used traditionally as a remedy for mouth and throat infections in Pushit community of Plateau state, Nigeria. Five solvents (hexane, chloroform, ethyl acetate, methanol and water) were used sequentially for the isolation of bioactive compounds from the root of the plant Adenodolichos paniculatus and concentrated on rotary vacuum evaporator. Concentrated extracts were tested for their antibacterial activities against Streptococcus pyogenes, Staphylococcus aureus, Pseudomonas aeruginosa and E. coli by agar-well diffusion method. Ofloxacin was the referenced drug. Chloroform root extract was more potent than the remaining four extracts. The extract at 100 mg/ml was active against only three (3) of the four (4) pathogens tested namely Streptococcus pyogenes, Staphylococcus aureus and Pseudomonas aeruginosa with zones of inhibition measuring 25.00, 21.00 and 11.75 mm respectively. The minimum inhibitory concentration of the extract against the test organisms were 1,560, 6,250 and 25,000 µg/ml respectively. The minimum bactericidal concentration was 3,120, 12,500 and 50,000 µg/ml against the three organisms. The extract was found to be nontoxic with LD50 of above 2000 (mg//kg body weight) when administered orally in mice. The plant extract has immunostimulatory activity with a significant increase (p<0.05) in white blood cell proliferation when administered into mice at doses of 50, 25 and 100 mg/kg. Conclusion: The result of this investigation supports the use of this plant for the traditional remedy of mouth and throat infections possibly caused by these test organisms. The extract can serve as lead in phytomedicine development for the treatment of throat related infections.

Fabrication of Hydroxypropyl Chitosan/Soy Protein Isolate Hydrogel for Effective Hemorrhage Control.

Hemostatic materials are increasingly important in civilian and military clinics. In this work, a hydrogel was fabricated from hydroxypropyl chitosan (HPCS) and soy protein isolate (SPI) through the crosslinking of epichlorohydrin. Effects of SPI content on the structure, and physical and biological properties of the prepared hydrogels were characterized using Fourier-transform infrared spectroscopy, X-ray diffractometry, scanning electron microscopy, water uptake testing, mechanical properties testing, MTT assay, hemolysis ratio testing, and routine blood coagulation test. The results indicated that the hydrogels showed high water uptake ability and compressive strength. The in vitro biocompatibility evaluation revealed that the hydrogel contains 30% SPI content (HCSH-30), could promote blood coagulation and cell proliferation. Furthermore, the hemostatic model of liver in New Zealand rabbit was applied to assess the hemostatic efficacy of the hydrogels. The results demonstrated that HCSH-30 stopped bleeding in 75 ± 1.63 s and improved hemostasis as compared with medical gauze. Thus, the HPCS/SPI hydrogel is expected to be a potential candidate for effective hemorrhage control. Impact statement Stoppage of bleeding is of paramount clinical significance in prophylactic, surgical, and emergency scenarios. This work describes a hydroxypropyl chitosan (HPCS)/soy protein isolate hydrogel, which could promote blood coagulation and cell proliferation, as well as stop bleeding in 75 ± 1.63 s on the liver of New Zealand rabbits. Thus, we provide a new candidate for hemostatic material and broaden the application of HPCS-based materials.

Abstract 6217: Synergistic effect of CDK8 and BCL-2 inhibition in AML through regulation of MCL-1 and BIM balance

Abstract Background Acute myeloid leukemia (AML) is characterized by rapid proliferation of myeloid blood cells. Due to its heterogeneity and the high rate of acquired drug resistance, new treatment modalities are needed. SEL120 is a specific type I selective inhibitor of Cyclin-dependent kinase 8 (CDK8) and Cyclin-dependent kinase 19 (CDK19). A first- in- human phase Ib clinical trial with SEL120 in patients with AML or HR-MDS was initiated in June 2019. Preclinical studies demonstrated high efficacy of SEL120 in experimental AML models via mechanisms involving differentiation and induction of apoptosis. Transcriptomic analysis of hematological cell lines demonstrated that SEL120 treatment upregulated expression of an apoptotic activator BIM from BCL-2 family of proteins. Methods Efficacy of the compound alone or in combination was tested in viability assays in a broad panel of cancer cell lines. Activity and mechanism of action of CDK8 inhibitor - SEL120 alone and in combination was investigated by flow cytometry, western blotting, co-immunoprecipitation, differential gene expression and ChIPseq analysis. In vivo efficacy was tested in mice injected with MV4-11 cell line both subcutaneously and intravenously. Results Here we provide a strong rationale for combination of SEL120 and BCL-2-selective inhibitor Venetoclax (ABT-199). We found that SEL120 synergistically induced apoptosis with Venetoclax in AML cells. Combination of both compounds significantly reduced levels of prosurvival protein MCL-1 and induced hallmarks of apoptosis including Caspase-3 activation and PARP cleavage. Previous studies associated Venetoclax resistance with increased sequestration of proapoptotic BIM by high levels of MCL-1. While a SEL120 treatment alone had no effects on MCL-1 levels, combination of both compounds resulted in sensitization of Venetoclax-resistant AML cells. We demonstrated that mechanism of Venetoclax resistance can be abrogated by the cotreatment with SEL120 leading to changes in proportions of BIM and MCL-1 levels. Synergistic interaction between SEL120 and Venetoclax resulted in apoptotic cell death in established cell lines and patient derived AML cells. Finally, using murine models of subcutaneous or disseminated AML, we found complete remissions of AML and associated recovery of normal cells in bone marrow of animals treated with both SEL120 and Venetoclax. Conclusion Taken together, these data provided rationale for a novel clinical strategy that may lead to durable responses in AML patients. Citation Format: Tomasz Rzymski, Marta Obacz, Milena Mazan, Marion Chappelier, Marcus Järås, Michal Mikula, Elżbieta Adamczyk, Katarzyna Wiklik, Michal Combik, Aniela Golas, Magdalena Masiejczyk, Przemyslaw Juszczynski, Jerzy Ostrowski, Krzysztof Brzózka. Synergistic effect of CDK8 and BCL-2 inhibition in AML through regulation of MCL-1 and BIM balance [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6217.

Co-microencapsulation of human umbilical cord-derived mesenchymal stem and pancreatic islet-derived insulin producing cells in experimental type 1 diabetes.

Post-partum umbilical cord Wharton Jelly-derived adult mesenchymal stem cells (hUCMS) hold anti-inflammatory and immunosuppressive properties. Human pancreatic islet-derived progenitor cells (hIDC) may de-differentiate, and subsequently re-differentiate into insulin producing cells. The two cell types share common molecules that facilitate their synergistic interaction and possibly crosstalk, likely useful for the cell therapy of type 1 diabetes (T1D). Upon microencapsulation in sodium alginate (AG), hUCMS and hIDC were able to form cell co-aggregates that looked well integrated and viable. We then grafted microencapsulated hUCMS/hIDC co-aggregates into non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice, and observed an acquired ability of cells to produce and store hormones. Finally, we transplanted these biohybrid constructs into NOD mice with recent onset, spontaneous overt diabetes, observing a decline of blood glucose levels. In vitro, we have shown that hUCMS inhibited proliferation of allogeneic polymorphonuclear blood cells from patients with T1D, while promoting expansion of FoxP3+ Tregs. Reversal of hyperglycemia in diabetic NODs seems to suggest that hUCMS and hIDC, upon co-microencapsulation, anatomically and functionally synergized to accomplish two goals: maintain tracer insulin output by hIDC, while exploting the immunoregulatory properties of hUCMS. We have gathered preliminary evidence that the two adult stem cell types within AG microcapsules, may synergistically promote tracer insulin production, while "freezing" the autoimmune disease process, and help reversal of the recent onset hyperglycemia in a spontaneous, autoimmune rodent model of diabetes, the NOD mouse, with no need for pharmacologic immunosuppression.

Investigation of Leukemogenic Mechanisms using Metabolomic Approaches

Leukemogenic mechanisms are characterized by the rapid proliferation of abnormal white blood cells that agglomerate in the bone marrow and subsequently interfere with the making of the normal blood cells. AML is a disparate neoplastic disorder that typified by naïve myeloid cell upsurge and bone marrow incompetence. AML is the most familiar type of leukemia affecting the aged. One prognostic feature associated with this response is the predominance of triggering mutations in FMS-like tyrosine kinase 3 (FLT3), especially the eventuality of internal tandem duplication (FLT3-ITD). AML with FLT3-ITD mutation is demonstrated to have a poor prognosis. Metabolomic profiling is a tool for understanding the modalities of the metabolites. In this study, we have attempted to understand AML with FLT3 from a metabolomic perspective. We have identified all the metabolites and their participating pathways, which are implicated in Acute myeloid leukemia (AML). AML is a hematological perturbation resulting from the proliferation and intensification of malignant myeloid cells. Formerly, smaller numbers of metabolites have been of use in the diagnosis of complex metabolic maladies and monogenic disorders like the inborn errors of metabolism. The results of this study enabled us to generate a metabolomic profile of FLT3/ITD patients, and we further analyzed the pathways from a network biology standpoint in which these metabolites are vital. We analyzed molecular networks with the help of Cytoscape and its plugins like Metscape. Arachidonic acid and Purine metabolism in cellular metabolites, while Glycolysis and Gluconeogenesis pathways, urea cycle and metabolism of arginine, proline, glutamate, aspartate, and asparagine metabolisms in plasma metabolites were the pathways having the highest impact in AML FLT3/ITD.

Ancient Homozygosity Segments in West African Djallonké Sheep Inform on the Genomic Impact of Livestock Adaptation to the Environment.

Simple SummaryAdaptation to the challenging environment of humid Sub-Saharan West Africa is hypothesized to cause an effect on the sheep genome. This can be used to identify genomic areas of importance to adaptation to changing environments. Djallonké sheep are small-sized, hair-coated trypano tolerant animals resulting from a unique process of natural adaptation. Here we identify ancient homozygous stretches, considered homozygous-by-descent, on the Djallonké sheep genome. Such genomic segments were assumed to be inherited from ancestors living during the historical time when sheep entered into West Africa and, therefore, are considered informative on the effect of human-mediated selection on the sheep genome. The genomic areas identified were involved in homeostasis and coagulation, innate immunity, defense against infections, white blood cells proliferation and migration, parasite resistance, and response to stress.A sample of Burkina Faso Djallonké (West African Dwarf) sheep was analyzed to identify stretches of homozygous segments (runs of homozygosity; ROH) overlapping with ancient homozygosity-by-descent (HBD) segments. HBD segments were considered ancient if they were likely to be inherited from ancestors living from 1024 to 2048 generations ago, roughly coinciding with the time in which sheep entered into West Africa. It is hypothesized that such homozygous segments can inform on the effect of the sheep genome of human-mediated selection for adaptation to this harsh environment. PLINK analyses allowed to identify a total of 510 ROH segments in 127 different individuals that could be summarized into 124 different ROH. A total of 32,968 HBD segments were identified on 119 individuals using the software ZooRoH. HBD segments inherited from ancestors living 1024 and 2048 generations ago were identified on 61 individuals. The overlap between consensus ROH identified using PLINK and HBD fragments putatively assigned to generations 1024 and 2048 gave 108 genomic areas located on 17 different ovine chromosomes which were considered candidate regions for gene-annotation enrichment analyses. Functional annotation allowed to identify six statistically significant functional clusters involving 50 candidate genes. Cluster 1 was involved in homeostasis and coagulation; functional clusters 2, 3, and 6 were associated to innate immunity, defense against infections, and white blood cells proliferation and migration, respectively; cluster 4 was involved in parasite resistance; and functional cluster 5, formed by 20 genes, was involved in response to stress. The current analysis confirms the importance of genomic areas associated to immunity, disease resistance, and response to stress for adaptation of sheep to the challenging environment of humid Sub-Saharan West Africa.

Acute lymphoid leukemia (LLA): A pediatric case report in dizygotic twins

Acute leukemias are considered neoplasias that reach the hematopoietic system, resulting in rapid clonal expansion and accelerated proliferation of blood cells. In this way, the cells lose their capacity for maturation and differentiation, accumulating young cells (blasts) and without function in the bone marrow. Acute lymphocytic leukemia (ALL) is a neoplasm that affects cells of the lymphoid lineage, but its prevalence occurs in B-cell precursors compared to T cells. It is common to occur in children between 1 and 5 years of age, comprising about 80% of the cases of all leukemias in this age group, but in adults, the probability of being present is lower. The present study aimed at the accomplishment of a case report on acute lymphoid leukemia in children, based on the comparison of risk factors in the family, and the evaluation of socioeconomic interference in the treatment, where, through an explanatory booklet, health professionals and the community about the major signs and symptoms of ALL in children. The research was submitted to the Ethics Committee of the Catholic University Center of Quixadá, through the Brazil Platform where it was analyzed and approved (01673218.2.0000.5046). In this study, we reported the case of two twin sisters who entered the Hospital São Sebastião de Pedra Branca, Ceará at different times, where one was admitted with fever and boulders, and another presenting only fever, after laboratory tests they were referred to a referral hospital in Fortaleza, Ceará, and were then diagnosed with acute lymphoid leukemia. Due to the poor prognosis of this pathology and a possible worsening of the patients' clinical status, they were referred for bone marrow transplantation. Fortunately, after the treatments performed, the twin had a positive evolution, no longer needing to undergo bone marrow transplantation

KCTD15 Protein Expression in Peripheral Blood and Acute Myeloid Leukemia.

Leukocytes are major cellular components of the inflammatory and immune response systems. After their generation in the bone marrow from hematopoietic stem cells, they maturate as granulocytes (neutrophils, eosinophils, and basophils), monocytes, and lymphocytes. The abnormal accumulation and proliferation of immature blood cells (blasts) lead to severe and widespread diseases such as leukemia. We have recently shown that KCTD15, a member of the potassium channel tetramerization domain containing protein family (KCTD), is remarkably upregulated in leukemic B-cells. Here, we extend our investigation by monitoring the KCTD15 expression levels in circulating lymphocytes, monocytes, and granulocytes, as well as in leukemia cells. Significant differences in the expression level of KCTD15 were detected in normal lymphocytes, monocytes, and granulocytes. Interestingly, we also found overexpression of the protein following leukemic transformation in the case of myeloid cell lineage. Indeed, KCTD15 was found to be upregulated in K562 and NB4 cells, as well as in HL-60 cell lines. This in vitro finding was corroborated by the analysis of KCTD15 mRNA of acute myeloid leukemia (AML) patients reported in the Microarray Innovations in Leukemia (MILE) dataset. Collectively, the present data open interesting perspectives for understanding the maturation process of leukocytes and for the diagnosis/therapy of acute leukemias.

Effect of Dietary Inorganic Copper on Growth Performance and Some Hematological Indices of Siberian Sturgeon Acipenser baerii Juveniles

AbstractThis research work was conducted to evaluate the effect of dietary copper (Cu) levels on growth performance, feed utilization, and some hematological parameters of juvenile Siberian Sturgeon Acipenser baerii. Five semipurified diets containing 0, 3, 6, 12, and 24 mg Cu/kg in the form of copper sulfate pentahydrate (CuSO4·5H2O) were formulated to provide the actual dietary copper values of 1.8, 5.7, 10.1, 15.9, and 28.3 mg Cu/kg diet, respectively. The obtained results exhibited a significant improvement in the final weight, protein efficiency ratio, daily growth coefficient, and hepatosomatic index with increasing dietary copper levels up to 10.1 mg/kg diet, while the feed conversion ratio significantly decreased. Also, erythrocyte count, hemoglobin, and hematocrit values increased with increasing levels of dietary copper, but no steady trend was observed in mean corpuscular hemoglobin changes. Furthermore, the neutrophils were significantly increased with enhancing the copper level in the fish diet, while the lymphocytes declined. Findings of the current research illustrate that an appropriate dietary copper level is required for improvement of growth performance, oxygen transfer capability, and white blood cell proliferation. Based on the broken‐line regression analysis of leukocyte count and monocyte percentage, the optimum dietary copper supplementation was estimated to be 10.63 and 10.36 mg Cu/kg diet, respectively.

Cell-free fat extract promotes tissue regeneration in a tissue expansion model

BackgroundTissue expansion techniques play an important role in plastic surgery. How to improve the quality of the expanded skin and shorten the expansion period are still worth investigating. Our previous studies found that a cell-free fat extract (CEFFE) possessed pro-angiogenic and pro-proliferative activities. However, the role of CEFFE on tissue expansion has remained unclear. The purpose of this study was to evaluate the effect of CEFFE on tissue expansion.MethodsA rat tissue expansion model was used. Animals were treated with CEFFE by subcutaneous injection. After 4 weeks of tissue expansion, the skin necrosis and retraction rates were evaluated, the thicknesses of the epidermis and dermis were determined by histological analyses, blood vessel density was measured by anti-CD31 staining, cell proliferation was assessed by proliferating cell nuclear antigen staining, and the expression of specific proteins was evaluated by western blot analyses. In addition, the effects of CEFFE on the proliferation and cell cycle of cultured HaCaT cells were evaluated in vitro.ResultsCEFFE treatment significantly decreased the necrosis rate and retraction of the expanded skin. The thickness of the epidermal and dermal layers was higher in CEFFE-treated compared to untreated skin. The density of blood vessels and cell proliferation in the epidermis of the expanded skin was improved by CEFFE treatment. In addition, CEFFE treatment significantly increased the expression of the vascular endothelial growth factor receptor, epidermal growth factor receptor, collagen type 1, and collagen type 3. CEFFE also increased the proliferation of HaCaT cells in culture.ConclusionsCEFFE improves the quality of the expanded skin by promoting angiogenesis and cell proliferation. It could be potentially used clinically for augmenting tissue expansion.

A Receptor Guanylate Cyclase, Gyc76C, Mediates Humoral, and Cellular Responses in Distinct Ways in Drosophila Immunity.

Innate immunity is an evolutionarily conserved host defense system against infections. The fruit fly Drosophila relies solely on innate immunity for infection defense, and the conservation of innate immunity makes Drosophila an ideal model for understanding the principles of innate immunity, which comprises both humoral and cellular responses. The mechanisms underlying the coordination of humoral and cellular responses, however, has remained unclear. Previously, we identified Gyc76C, a receptor-type guanylate cyclase that produces cyclic guanosine monophosphate (cGMP), as an immune receptor in Drosophila. Gyc76C mediates the induction of antimicrobial peptides for humoral responses by a novel cGMP pathway including a membrane-localized cGMP-dependent protein kinase, DG2, through downstream components of the Toll receptor such as dMyD88. Here we show that Gyc76C is also required for the proliferation of blood cells (hemocytes) for cellular responses to bacterial infections. In contrast to Gyc76C-dependent antimicrobial peptide induction, Gyc76C-dependent hemocyte proliferation is meditated by a small GTPase, Ras85D, and not by DG2 or dMyD88, indicating that Gyc76C mediates the cellular and humoral immune responses in distinct ways.

First expression and immunogenicity study of a novel trivalent outer membrane protein (OmpII-U-A) from Aeromonas hydrophila, Vibrio vulnificus and Edwardsiella anguillarum

Aeromonas hydrophila, Vibrio vulnificus and Edwardsiella anguillarum are three common bacterial pathogens in cultivated eels. To protect farming eels from infection by these pathogens, a trivalent outer membrane protein (OmpII-U-A) containing partial sequences of OmpII from A. hydrophila, OmpU from V. vulnificus and OmpA from E. anguillarum was expressed as a vaccine to immune European eels (Anguilla anguilla). The expression of 4 immune-related genes, proliferation of whole blood cells, antibody titers in serum and mucus, and activity of SOD were detected at different time after the immunization, and the relative percent of survival (RPS) was determined after eels were infected by A. hydrophila, V. vulnificus or E. anguillarum at 28 d post the immunization (dpi). The results showed that the OmpII-U-A could significantly stimulate the gene expression of sod, mhc2 and lysC post the immunization; SOD activity and the antibody titer in serum and skin mucus were significantly increased. After the challenge on 28 dpi, the RPSs of OmpII-U-A against A. hydrophila, V. vulnificus and E. anguillarum were 66.67%, 44.44% and 22.22%, respectively. The results showed that the OmpII-U-A could protect eel from A. hydrophila and V. vulnificus infection, but had weak protection effect on E. anguillarum in European eels. Expression and immunogenicity of a trivalent Omp were reported for the first time, and results of this study will provide valuable reference for the development of fish multiplex vaccine.

Immunization of a novel bivalent outer membrane protein simultaneously resisting Aeromonas hydrophila, Edwardsiella anguillarum and Vibrio vulnificus infection in European eels (Angullia angullia)

In cultivated European eels, Aeromonas hydrophila, Edwardsiella anguillarum and Vibrio vulnificus are three important bacterial pathogens. In this study, European eels (Anguilla anguilla) were immunized by the bivalent expression products of the outer membrane protein (Omp) gene from A. hydrophila (OmpⅡ) and E. anguillarum (OmpA), and the effects of the bivalent protein (rOmpⅡ-A) on the immune function of the European eel were detected. Three hundred eels were divided average into three groups of PBS, adjuvant and rOmp. Eels of three goups were injected intraperitoneal with 0.2 mL of PBS (0.01 mol/L, pH7.4), PBS + F (PBS mixed equal volume of freund's uncomplete adjuvant) or rOmpⅡ-A (1 mg mL−1 rOmpⅡ-A mixed equal volume of freund's uncomplete adjuvant). Four immune-related genes expression, proliferation of whole blood cells, serum and skin mucus antibody titer, superoxide dismutase (SOD) activity and the relative percent of survival (RPS) were studied at different days (or hours) post the immunization. The results showed that the igm, lysC, mhc2 and sod gene in the liver, spleen, kidney and intestine tract were significant increased in the Omp group; On the 28 day post the immunization (dpi), blood cell proliferation was increased in the Omp group, and on the 14, 21, 28 and 42 dpi, antibody titers in serum and mucus of the Omp group were significantly higher than that of the PBS and adjuvant group, regardless of coating with bacteria or Omp antigen. The SOD activity of Omp group increased significantly in liver, kidney, skin mucus and serum from 14 to 42 dpi, especially in serum. Eels chanllenged by A. hydrophila, E. anguillarum and V. vulnificus in the bivalent Omp group showed the RPS were 83.33%, 55.56% and 44.44%, respectively. The results of this study showed that immunization of the bivalent Omp could effectively improve the immune function of European eels, and produced effectively protection to A. hydrophila and E. anguillarum infection. Simultaneously, the bivalent Omp also produced distinct cross-protection to the eels challenged by V. vulnificus.

Integrating Enhancer Profiling and CRISPR Dropout Screen Revealed Selenoprotein Synthesis Pathway As a New Vulnerability in AML

Alterations of the epigenetic landscape and transcription are hallmarks of acute myeloid leukemia (AML) that drive leukemogenic gene expression and therefore can be exploited for therapeutic intervention. To look for such targets that harbor both an altered epigenetic feature and are genetically essential for AML cells, we performed a multi-database analysis integrating pan-cancer super enhancer landscapes with whole genome CRISPR dropout screens. Among the top targets, we discovered SEPHS2. An enhancer was present upstream of the gene marked by H3K27ac and bound by leukemogenic transcription factors including MYB, Pu.1 and RUNX1. In addition, AML cells with SEPHS2 deletion significantly dropped out in a genome wide CRISPR screen. This gene encodes a critical enzyme in the underappreciated selenoprotein synthesis pathway which was highly upregulated in TCGA AML patients compared to control blood cells from healthy individuals. Collectively, our initial bioinformatic analysis suggested that the selenoprotein synthesis pathway is a new vulnerability in AML. To test the functional requirement of the selenoprotein synthesis pathway in AML and other cells, we performed CRISPR mediated deletion of three key genes in the selenoprotein synthesis pathway, SEPHS2, SEPSECS and EEFSEC. The human AML cell lines (MOLM13, THP1 and Kasumi1), murine AML cells transformed by MLL-AF9 and human AML PDX cells all depicted a significant dependency on these genes, while proliferation of normal cord blood cells and myeloma cells (U266B1) was almost not affected. We then transplanted these cells into recipients. Deletion of SEPHS2, SEPSECS or EEFSEC significantly ameliorated AML progression, indicated by decreased AML burden and extended survival. Since selenoproteins including GPX1 and GPX4 are known antioxidants, we hypothesized that perturbing the selenoprotein synthesis pathway disrupted the redox state in AML cells. We found that deletion of SEPHS2, SEPSECS or EEFSEC elevated ROS in AML cells as demonstrated by Cell-Rox or DCF-DA staining. Western blotting revealed significantly downregulated GPX4 level and upregulated DNA damage marker γ-H2AX. Moreover, the defective proliferation was partially rescued by adding antioxidant TEMPOL. These results suggested selenoprotein synthesis pathway produced key antioxidants to balance the proper redox state and was required for AML cell proliferation. A major source of selenium is diet. Therefore, we hypothesized that consuming selenium low diet could suppress AML. We compared the survival of AML bearing mice on selenium proficient and deficient diet. The selenium deficient diet significantly extended survival, lowered GPX4 level and increased ROS in AML cells. Interestingly, normal mouse on selenium deficient diet for a 3-months period did not develop any abnormalities in CBC or bone marrow hematopoiesis. This suggests selenium deficient diet could be clinically applicable without significant side effects. Altogether, the integration of a pan-cancer enhancer landscape study with CRISPR dropout gene screen offered a powerful tool to dissect cancer targets that possessed unique enhancer features and genetic essentiality. The analysis yielded SEPHS2 and its selenoprotein synthesis pathway to be a new vulnerability in AML. The underappreciated selenoprotein synthesis pathway was key to produce antioxidant selenoproteins such as GPX1 and GPX4 to maintain a proper redox state in AML. Deleting the genes or removing selenium from diet could perturb the pathway and ameliorate the AML disease. Disclosures Lin: Syros Pharmaceuticals: Equity Ownership, Patents & Royalties.

Efek Ekstrak Petroleum Eter Daun Ara (Ficus Racemosa Linn.) pada Indeks Fagositosis Mencit (Mus musculus)

Senyawa terpenoid daun ara diketahui memiliki aktivitas imunosupresan dengan menghambat proliferasi sel darah. Imunosupresan merupakan golongan obat yang dapat menekan respon imun berlebihan. Penelitian ini bertujuan untuk mengetahui aktivitas imunosupresan ekstrak petroleum eter daun ara (Ficus racemosa. Linn) terhadap mencit (Muss musculus). Uji imunosupresan dilakukan dengan metode bersihan karbon untuk mengetahui kemampuan fagositosis dengan karbon sebagai antigen yang diinjeksikan intravena, dilanjutkan dengan menghitung indeks organ. Seluruh data yang diperoleh kemudian dianalisis statistik menggunakan SPSS. Hasil uji bersihan karbon menunjukkan bahwa ekstrak petroleum eter daun ara dosis 560 mg/kgBB bersifat imunostimulan sedang dosis 1120 mg/kgBB bersifat imunostimulan lemah dengan nilai indeks fagositosis berturut-turut 1,5 dan 1,1, sedangkan dosis 1680 mg/kgBB bersifat imunosupresan dengan indeks fagositosis 0,7. Hasil uji indeks organ limfoid menunjukkan perbedaan signifikan antara ekstrak petroleum eter dosis 1680 mg/kgBB dengan kontrol negatif (p<0.05). Berdasarkan hasil uji respon imun non spesifik disimpulkan bahwa ekstrak petroleum eter daun ara dosis 1680 mg/kgBB mempunyai aktivitas imunosupresan.

The PNPLA3 I148M variant promotes lipid-induced hepatocyte secretion of CXC chemokines establishing a tumorigenic milieu.

The I148M variant of the Patatin-like phospholipase domain-containing 3 (PNPLA3) protein is associated with an increased risk for liver inflammation and hepatocellular carcinoma (HCC), but the underlying mechanism is unknown. We hypothesized that enhanced CXC chemokine secretion mediates hepatic inflammation that accelerates development of HCC. Expandable primary human (upcyte®) hepatocytes and human PLC/PRF/5 hepatoma cells were lentivirally transduced with both PNPLA3 I148M variants and stimulated with lipids. Cytokine levels in culture supernatant and patient sera (n = 80) were analyzed by ELISA. Supernatants were assessed in transmigration experiments, tube formation, and proliferation assays. In vitro, lipid stimulation of transduced hepatocytes dose-dependently induced the production of interleukin-8 and CXCL1 in hepatocytes carrying the PNPLA3 148M variant. In line, sera from PNPLA3 148M-positive patients with alcoholic liver cirrhosis contained higher levels of interleukin-8 and CXCL1 than patients with wild-type PNPLA3. Supernatants from lipid-stimulated hepatocytes with the PNPLA3 148M variant induced enhanced migration of white blood cells, angiogenesis, and cell proliferation in comparison with supernatants from wild-type hepatocytes via CXC receptors 1 and 2. Increased production of interleukin-8 and CXCL1 by hepatocytes carrying the PNPLA3 148M variant contributes to a pro-inflammatory and tumorigenic milieu in patients with alcoholic liver disease. KEY MESSAGES: The PNPLA3 148M variant is associated with cirrhosis and hepatocellular carcinoma. Lipid stimulation of hepatocytes with this variant induces IL-8 and CXCL1. Supernatants from hepatocytes with this variant promote migration and angiogenesis. Sera from patients with this variant contained enhanced levels of IL-8 and CXCL1. The PNPLA3 148M variant contributes to a tumorigenic milieu via IL-8 and CXCL1.

Design of Drug Molecule Based Structure for Smad 3 Protein and Its Insilico Research of Leukemia Utilizing Bioinformatics Software

The Research work on rational drug molecules design for Smad3,the conclusion is that out of all the inhibitors chosen for the docking , PTHrP emerged to be the best inhibitor for the protein Smad3.The precise reason for it was its docking energy which was the lowest (docked energy =-29.73), among all other inhibitors used. Hence the conclusion is that Smad3 is the best inhibitor, for Smad3. Hence the task was completed successfully. There are many reasons and factors responsible for induction of cancer. Leukemia is a type of cancer in blood or bone marrow and is characterized by an irregular proliferation of white blood cells.

Acute multiple cerebral infarction combined with cerebral microhemorrhage in Polycythemia vera: A case report.

Polycythemia vera (PV) is one of the rare causes of cerebrovascular disease, whose common manifestations in the nervous system are cerebral infarction and transient ischemic attack. A number of cases of PV patients with bleeding complicated with subdural hemorrhage or cerebral hemorrhage have been previously reported. However, sometimes PV patient with complicated cerebral hemorrhage and lower extremity venous thrombosis have been admitted to the People's Hospital of Liaoning Province. The present case study reports on a patient with acute multiple cerebral infarction with cerebral micro-hemorrhage associated with PV, who was not treated with anti-thrombosis treatment. After bloodletting treatment and hydroxyurea treatment, the patients condition was stable and they were discharged. A possible mechanism of infarction is that PV may cause abnormal proliferation of red blood cells, white blood cells and platelets in the circulation, resulting in an increase of blood viscosity and reduction of blood flow velocity. Platelet deficiency may cause abnormal blood coagulation function, which may be the reason for the blood and thrombotic diseases in patients with PV.

SETDB1 modulates the differentiation of both the crystal cells and the lamellocytes in Drosophila

Proper genetic and epigenetic regulation is necessary to maintain the identity and integrity of cells. Enzymes involved in post-transcriptional modifications of histones are key factors in epigenetic mechanisms. Such modifications are also gaining importance for their role in growth and development of cancer. SETDB1 catalyzes the epigenetic mark of lysine-9 methylation of histone-3. In this study, we explored the role of SETDB1 in Drosophila hematopoiesis. We show that SETDB1 controls the differentiation of matured blood cells in wandering third instar larvae. There are three matured blood cells in wild type Drosophila melanogaster: plasmatocytes, crystal cells and lamellocytes. We found that loss-of-function mutants of SETDB1 show hematopoietic defects; increased blood cell proliferation, decreased number of crystal cells, greater differentiation of blood cells into lamellocytes, dysplasia of the anterior lobes of lymph gland and presence of hematopoietic tumors. Cell type specific knockdown of SETDB1 provided similar phenotype i.e., decreased number of crystal cells and an increase in lamellocyte differentiation. In animals with loss of function of SETDB1, Notch pathway was downregulated. Further, over-expression of SETDB1 in blood cells resulted in an increase in the number of crystal cells. This increase is accompanied with an increase in the number of NotchICD expressing cells. We therefore performed genetic rescue using UAS-GAL4 system to rescue loss of function SETDB1 mutants. Our data show that the rescued larvae carrying a wild type copy of SETDB1 in mutant background are devoid of blood tumors. We have identified a novel dual function of SETDB1 methylatransferase as a critical regulator of two of the matured hemocytes, crystal cells and lamellocytes. We propose a novel role of SETDB1 in modulating the differentiation of crystal cells and lamellocytes from a common progenitor and underscore the importance of SETDB1 in Drosophila blood tumor suppression.