scholarly journals Evaluation of Antibacterial, Antineoplastic, and Immunomodulatory Activity ofPaullinia cupanaSeeds Crude Extract and Ethyl-Acetate Fraction

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Lidiane Vasconcelos do Nascimento Carvalho ◽  
Marina Ferraz Cordeiro ◽  
Thiago Ubiratan Lins e Lins ◽  
Maria Clara Pinheiro Duarte Sampaio ◽  
Gabriela Souto Vieira de Mello ◽  
...  
Keyword(s):  
Ethyl Acetate ◽  
Crude Extract ◽  
Biological Activities ◽  
Leukemia Cell ◽  
B Cell Lymphoma ◽  
Ethyl Acetate Fraction ◽  
Leukemia Cell Line ◽  
Breast Adenocarcinoma ◽  
Immunomodulatory Activity ◽  
Native Plant

Paullinia cupana(Guarana) is a native plant of Amazon region that has very traditional importance. Its seeds are rich in bioactive compounds, including tannins, which exhibit relevant properties.Objective.This study aimed to evaluate antibacterial, antineoplastic, and immunomodulatory activity ofP. cupanaseeds crude extract (CE) and ethyl-acetate fraction (EAF).Methods.Antibacterial activity was evaluated by determination of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). Antineoplastic activity was evaluated by MTT assays in hepatocellular carcinoma (HepG2), breast adenocarcinoma (MCF-7), ductal carcinoma (T47-D), non-Hodgkin’s B cell lymphoma (Toledo), T cell leukemia (Jukart), and Acute Leukemia (HL-60) cell lines. BALB/c mice splenocytes were treated to assess IFN-γ, IL-6, IL-17, and IL-10 levels by sandwich ELISA.Results.CE and EAF were not toxic to peripheral blood cells and splenocytes. CE and EAF fractions showed a bacteriostatic activity (MIC = 250 μg/mL) and presented IC50values of 70.25 μg/mL and 61.18 μg/mL in HL-60 leukemia cell line. All cytokines evaluated had their levels reduced after treatment, following dose-response model.Discussion and Conclusion.Different biological activities were observed for both CE and EAF, suggestingP. cupanaas a source of bioactive substances, especially tannins that may be used for several diseases treatments.

scholarly journals CYTOTOXIC BIOACTIVE COMPOUNDS FROM CALOTROPIS GIGANTEA STEM BARK

2016 ◽  
Vol 8 (9) ◽  
pp. 111
Author(s):  
Kartini Hasballah ◽  
Murniana . ◽  
Erya . ◽  
Ardian .
Keyword(s):  
Ethyl Acetate ◽  
Cytotoxic Activity ◽  
Ethyl Acetate Extract ◽  
Leukemia Cell ◽  
Stem Bark ◽  
Hexane Extract ◽  
Leukemia Cell Line ◽  
Calotropis Gigantea ◽  
Murine Leukemia Cell ◽  
Main Components

<p><strong>Objective: </strong>The present study deals with the cytotoxic activity of n-hexane and ethyl acetate extracts of <em>Calotropis gigantea</em> L. stem bark and its fractions such as A, B, C, D and E fractions on murine leukemia cell line P388.</p><p><strong>Methods: </strong>The crude extracts of <em>C. gigantea</em> stem bark were prepared using n-hexane and ethyl acetate solvents. The plant extracts were subjected to vacuum liquid chromatography followed by TLC. According to the similarity of stain patterns, the fractions were combined. The extracts and its combined fractions were then subjected for the phytochemical test. Cytotoxic activity of those extracts and its combined fractions were tested using MTT assay. Fraction D was subjected to gravity column chromatography followed by TLC. Then, fractions A, B, and D2 were crystallized and subjected to GC-MS.</p><p><strong>Results: </strong>The qualitative screening of n-hexane extract of <em>Calotropis gigantea</em> L. stem bark for secondary metabolites showed the presence of terpenoid, flavonoids, phenolics and coumarins. While the ethyl acetate extract contained phenolics, steroids, flavonoids, saponins and coumarins compounds. IC<sub>50 </sub>values for n-hexane extract and E fraction are 76.29 µg/ml and 18.48 µg/ml, respectively. In the ethyl acetate extract and C fraction obtained IC<sub>50</sub> values 57.05 µg/ml and 52.58 µg/ml.</p><p><strong>Conclusion: </strong>Cytotoxic activity from E fraction of n-hexane extract of <em>C. gigantea</em> stem bark is the most potent and containing flavonoids, phenolics and coumarins. The main components from several compounds of n-hexane extract of <em>C. gigantea</em> are germacrane-A, (-)-globulol, urs-12-ene and veridiflorol. </p>

Exploring cytotoxic and antioxidant properties of Heliotropium calcareum in polar and non-polar extracts

2021 ◽  
Vol 02 ◽  
Author(s):  
Mohammad Uzair ◽  
Faisal Rashid ◽  
Hamid Saeed Shah ◽  
Jamshed Iqbal
Keyword(s):  
Cell Cycle ◽  
Ethyl Acetate ◽  
Hela Cells ◽  
Biological Activities ◽  
Research Work ◽  
Antioxidant Properties ◽  
Ethyl Acetate Fraction ◽  
Cell Cycle Analysis ◽  
G1 Phase ◽  
Traditional Use

Background: Plants are a vital source of natural drugs as the traditional use of plants as therapeutic agents for a variety of ailments has been traced back to thousands of years. The utilization of Heliotropium calcareum has been evident since ancient times for treating various disease states like inflammation associated with gout and rheumatism, poisonous bites, and other skin disorders. The current research work was carried out to determine the phytochemistry and biological activities of the crude methanolic extract obtained through maceration from the aerial parts of Heliotropium calcareum. Methods: The plant was collected from district Bhakkar, Punjab, Pakistan. Maximum phenolic (74.5 µg GAE/mg) and flavonoid content (58.99 µg QE/mg) were observed in ethyl acetate fraction. Significant antioxidant potential was observed in ethyl acetate fraction with the highest free radical hunting activity of 92.6 ± 6.7 µM. Results: Cytotoxicity assay using MTT dye was performed where non-polar (n-hexane) and polar (ethyl acetate) fractions displayed excellent cytotoxicity against HeLa cells (IC50 = 79.95 ± 3.718 & 164 ± 4 µg/mL respectively). Furthermore, the above fractions showed momentous results in cell cycle analysis and promising proapoptotic effect against cervical (HeLa) cancer cell lines. An n-hexane and ethyl acetate fraction were selected for cell cycle analysis to determine the quantitative measurement of the degree of apoptosis. According to the results given below in the figure, the cervical (HeLa) cancer cells were treated with n-hexane and ethyl acetate fractions at various concentrations. An increase in the cell population at G0/G1 phase and a decrease in the S-phase population concerning untreated cells suggested the G0/G1 phase arrest in n-hexane and ethyl acetate fractions treated HeLa cells. Conclusion: Overall, , n-hexane and ethyl acetate fractions were found to be the most promising and active elements of H. calcareum and may be utilized to explore their cytotoxic effects further in the animal model.

scholarly journals Study on biological activities of extracted fractions from Typhonium flagelliforme (Lodd.) Blume

2017 ◽  
Vol 33 (2S) ◽  
Author(s):  
Le Quy Thuong ◽  
Bach Tuyet Mai ◽  
Nguyen Minh Chau ◽  
Le Thi Phuong Hoa ◽  
Nguyen Quang Huy
Keyword(s):  
Ethyl Acetate ◽  
Cytotoxic Activity ◽  
Biological Activities ◽  
Ethyl Acetate Fraction ◽  
Positive Control ◽  
Chemical Compositions ◽  
Active Components ◽  
Stomach Pain ◽  
Ic50 Values

Typhonium flagelliforme is a medicinal plant that has variety of uses. In medicinal traditional T. flagelliforme is used to treatment cough, headache, stomach pain chronic, and tracheitis. Moreover, use fresh bulbs treatment furuncle, the bites of poisonous insects. The active components in T. flagelliforme are flavonoids. In this study, the T. flagelliforme extract was obtained by methanol  to determine the chemical composition. Then, The extracts of methanol are extracted with polarization increases gradually solvents such as haxane, dichloromethane and ethyl acetate. Determination of antioxidant activity, cytotoxic activity of extracted fractions. Results obtained showed that the chemical compositions by the qualitative reaction preliminary were identified from T. flagelliforme containing reducing sugars, amino acids, organic acids, flavonoids, alcaloids, sterols. The antioxidant capacity of the ethyl acetate fraction reached 94.76 μg/ml, 10 times higher than the positive control is Quercetin. Cytotoxic activity of the haxane and diclomethane extracted fractions from T. flagelliforme exhibited cytotoxic activity on all three experimental cancers cell lines: KB, HepG2, Lu after 72h of culture with IC50 values ​​range from 92.8 to 107.76 μg/ml. From dichlomethane extracted of T. flagelliforme was purified TF1 as Stigmast-4-en- 3-on.    

scholarly journals Phytochemical, antimicrobial, antioxidant and enzyme inhibitory potential of medicinal plant Dryopteris ramosa (Hope) C. Chr.

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Fiaz Alam ◽  
Syed Hurmat Ali Khan ◽  
Mohammad Hassham Hassan Bin Asad
Keyword(s):  
Ethyl Acetate ◽  
Crude Extract ◽  
Radical Scavenging ◽  
Ethyl Acetate Fraction ◽  
Phytochemical Analysis ◽  
Methanol Fraction ◽  
Medicinal Uses ◽  
Antibiotic Effect ◽  
Esterase Inhibition ◽  
Mic Value

Abstract Background Dryopteris ramosa has numerous potentials uses in the treatment of different maladies as old traditional medication. The fronds of D. ramose are edible and orally administered for producing antibiotic effect. They are also used as astringent and febrifuge, and as a pesticide. Methods Extraction of fronds of D. ramosa using solvents of increasing polarity, namely, ethyl acetate, methanol and water were tested for phytochemical (qualitative tests, GC-MS), antimicrobial (well method), antioxidant (DPPH), antifungal (tube dilution), cytotoxic activity (brine shrimps lethality assay) and LOX and COX inhibitory activities were performed using standard methods. Results The phytochemical analysis of the crude methanolic extract revealed that the fronds are rich in flavonoids, alkaloids, saponins, tannins, glycosides and triterpenoids. The total flavonoid content of the ethyl acetate fraction was 46.28 μg QE/mg extract. The GC-MS analysis revealed nine major compounds that constituted the crude drug and potentially had a role in reported activities. The crude extract was the most active amongst all the fractions against the bacterial and fungal strains used such that it inhibited the growth of P. aeruginosa with a zone of 13 mm and a MIC value of 16 μg/ml as compared to the standard cefixime, which inhibited the zone by only 10 mm and a MIC value of 32 μg/ml. The highest antioxidant potential in DPPH assay was shown by the crude extract with 91.948% free radical scavenging activity. The bring shrimps lethality potential of the crude extract was the highest, with a LD50 value of 47.635 μg/ml. The ethyl acetate fraction inhibits 91.36% of alpha glucosidase enzyme at a concentration of 0.5 mg/ml. In case of acetylcholine esterase inhibition assay, the methanol fraction inhibits 58.26% of the enzyme activity. Similarly, for butyrylcholine esterase inhibition, the maximum inhibitory effect was seen in the methanol fraction, with a percentage inhibition of 47.32%. Conclusion These test results support traditional medicinal uses of the plant. Dryopteris ramosa could be imperative for being used as a therapeutic agent and the medicinal importance of this plant should be further investigated.

scholarly journals Reduksi Udema Kaki Mencit (Mus musculus) Terinduksi Karagenan Setelah Pemberian Ekstrak Daruju (Acanthus ilicifolius L.)

2020 ◽  
Vol 17 (2) ◽  
pp. 86-91
Author(s):  
Darul Ikhwan ◽  
Octavian Rachmadi ◽  
Annisa Dyah Astari Putri ◽  
Ari Widiyantoro
Keyword(s):  
Secondary Metabolites ◽  
Ethyl Acetate ◽  
Mus Musculus ◽  
Crude Extract ◽  
Antiinflammatory Activity ◽  
Raw Materials ◽  
Ethyl Acetate Fraction ◽  
West Kalimantan ◽  
Mangrove Plants ◽  
Acanthus Ilicifolius

Acanthus ilicifolius L. is one of the mangrove plants that contain a lot of potential secondary metabolites of medicinal raw materials. People of Sambas Regency of West Kalimantan Province uses leaves extract daruju to overcome the pain in the stiff and swollen. The objective of this study was to know the antiinflammatory activity of leaves extract of daruju, in the form of edema reduction of mices feet induced carrageenan. The method used in the form of maceration and partition extraction followed by phytochemical test of crude extract and the fraction of result of partition. Furthermore, the crude extract and the fraction of the result of the partition were tested for the reduction of edema mices feet induced carrageenan. The results showed that ethyl acetate fraction had the best antiinflammatory activity because it was able to decrease the edema of mices feet by 75% at 750 mg / kg BW. Ethyl acetate fraction has the ability to reduce edema because it contains group compounds of flavonoid and terpenoid.

scholarly journals Immunomodulatory Activity of Phyllanthus maderaspatensis in LPS-Stimulated Mouse Macrophage RAW 264.7 Cells

Separations ◽  
2021 ◽  
Vol 8 (9) ◽  
pp. 129
Author(s):  
Uoorakkottil Ilyas ◽  
Deepshikha P. Katare ◽  
Punnoth Poonkuzhi Naseef ◽  
Mohamed Saheer Kuruniyan ◽  
Muhammed Elayadeth-Meethal ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Liquid Chromatography ◽  
Ethyl Acetate ◽  
Ethyl Acetate Fraction ◽  
Raw 264.7 Cells ◽  
Immunomodulatory Activity ◽  
No Production ◽  
Raw 264.7 ◽  
Mouse Macrophage ◽  
Significant Stimulatory Effect

Phyllanthus species (Family Euphorbiaceae) has been used in traditional medicine of several countries as a cure for numerous diseases, including jaundice and hepatitis. This study is an attempt to evaluate the immunomodulatory activity of various fractions, column eluents of ethyl acetate fraction, and their polyphenols. Phyllanthus maderaspatensis were standardized using high-performance liquid chromatography to identify and quantify polyphenols, and purification of polyphenols was carried out using vacuum liquid chromatography. Subsequently, we tested various fractions, column eluents of ethyl acetate fraction, and polyphenols in vitro to assess their impact on nitric oxide (NO) production in LPS-stimulated mouse macrophage RAW 264.7 cells. The ethyl acetate fraction (100 μg mL−1) had a more significant stimulatory effect on LPS-stimulated NO production by the RAW 264.7 cells. We found that the ethyl acetate fraction contains a high amount of catechin, quercetin, ellagic acid kaempferol, and rutin, which are responsible for immunomodulation. The ethyl acetate fraction at concentrations of 25 and 50 μg mL−1 had a significant inhibitory effect and 100 μg mL−1 had a more significant stimulatory effect when compared with the LPS control. The percentage of inhibition by LPS control ranged from zero percentage, kaempferol ranged from 45.4% at 50 μg mL−1 to 41.88% at 100 μg mL−1, catechin ranged from 50% at 50 μg mL−1 to 35.28% at 100 μg mL−1, rutin ranged from 36.2% at 50 μg mL−1 to 47.44% at 100 μg mL−1, gallic acid ranged from 28.4% at 50 μg mL−1 to 50.9% at 100 μg mL−1, ellagic acid ranged from 45.12% at 50 μg mL−1 to 38.64% at 100 μg mL−1, and purified quercetin ranged from 26.2% at 50 μg mL−1to 45.48% at 100 μg mL−1. As NO plays an important role in the immune function, polyphenols’ treatment could modulate several aspects of host defense mechanisms owing to the stimulation of the inducible nitric oxide synthase.

scholarly journals Immunomodulatory Activity on Specific Immune Function in Albino Mice Exposed to Extract and Fractions of Zapoteca portoricensis (Jacq) HM. Hernández Roots

2021 ◽  
pp. 41-50
Author(s):  
Oghenesuvwe D. Warren ◽  
Bonaventure C. Obi ◽  
Uchenna C. Abonyi ◽  
Theophine C. Akunne
Keyword(s):  
Ethyl Acetate ◽  
Normal Control ◽  
Ethyl Acetate Fraction ◽  
Immunomodulatory Activity ◽  
Neutrophil Adhesion ◽  
Root Extract ◽  
Percentage Increase ◽  
Methanol Fraction ◽  
Group I ◽  
Study Results

Zapoteca portoricensis (Jacq) HM. Hernández popularly called “Elugelu” belongs to the Fabaceae family. It is reportedly used in traditional medicine as anti-diarrhoea, anticonvulsant, antispasmodic and in management of autoimmune disorders. The immunomodulatory activity of methanol root extract and fractions of Zapoteca portoricensis was evaluated using three experimental models: neutrophil adhesion, haemagglutination antibody (HA) titre and delayed hypersensitivity (DTH) of an immune response in mice. Five (5) groups of mice (n=5) were used; group I was the normal control; group II was the standard control (levamisole, 2.5 mg/kg) while groups IIIA–C, IVA–C and VA–C received graded doses (25, 50, 100 mg/kg) of methanol extract (ME), ethyl acetate fraction (EF) and methanol fraction (MF), respectively. Sheep red blood cells (SRBCs; 0.1 ml) were injected subcutaneously to sensitize the animals. The study results showed that the methanol fraction (MF) exhibited the highest percentage (46.12%) in neutrophil adhesion followed by the ethyl acetate fraction (EF) (37.06%) at 100 mg/kg dose, respectively, compared with the normal control. The highest percentage increase in both primary and secondary antibody titre was found to be ME (85.19%, 43.20%), EF (91.53%, 102.67%) and MF (128.31%, 111.89%) at 50 mg/kg dose, respectively compared to the normal control. The EF at the doses of 25 and 50 mg/kg produced the highest percentage inhibition (56.57%, 58.33%) in DTH response, respectively, followed by MF (42.46%) at a dose of 25 mg/kg. Oral administration of Z. portoricensis exhibited immunomodulatory effects on specific components of the immune system in mice.

scholarly journals PENENTUAN POTENSI EKSTRAK KULIT BATANG TUMBUHAN SIKKAM (Bischofia javanica Blume) SEBAGAI ANTIOKSIDAN DENGAN METODE DPPH DAN SITOTOKSIK DENGAN METODE BSLT

Alotrop ◽  
2020 ◽  
Vol 4 (1) ◽  
Author(s):  
Dapot Parulian Manurung ◽  
Agus Sundaryono ◽  
Hermansyah Amir
Keyword(s):  
Antioxidant Activity ◽  
Ethyl Acetate ◽  
Crude Extract ◽  
Anticancer Agent ◽  
Ethyl Acetate Fraction ◽  
Hexane Fraction ◽  
Bark Extract ◽  
Test Results ◽  
Bischofia Javanica ◽  
Ethanol Fraction

This study aims to determine the level of antioxidant activity of Sikkam bark extract (Bischofia javanica Blume) and its potential as an anticancer agent. This research was conducted in November 2018 - April 2019 in the Laboratory of Chemical Education Guidance and Counseling and Basic Science Laboratory, University of Bengkulu. The sample in this study was obtained from Gunung Mariah village in Simalungun district, North Sumatra. The method used in this study was extraction using maceration, liquid-liquid fractionation, phytochemical test, DPPH (Diphenylpicrylhiydrazyl) test and BSLT test (Brine Shrimp Lethality Test). Extraction maceration using ethanol 96%, fractionation using a solvent n-hexane and ethyl acetate, phytochemical tests using Mayer's reagent, Lieberman-Burchard, FeCl3 1% and H2SO4 2N, antioxidant test using DPPH (1,1-diphenyl-2-picrylhydrazyl) and cytotoxic tests using shrimp Artemia salina Leach larvae. The results of the study on maceration of 800 grams of Sikkam bark obtained a crude extract of 53.6 grams (6.7%). Fractionation from 10 grams of crude extract obtained 7.24 gram ethanol fraction (72.4%), ethyl acetate fraction 2.14 gram (21.4%) and n-Hexane fraction 0.39 gram (3.9%). Phytochemical test results obtained the content of alkaloids, flavonoids, tannins, saponins and terpenoids with the highest yield on tannin compounds. The antioxidant activity test results obtained IC50 values in crude extracts of 20.94 ppm, ethanol fraction 36.29 ppm, ethyl acetate fraction 83.28 ppm and n-Hexane fraction 39.13 ppm which showed that crude extract, ethanol fraction, n-Hexane fraction were categorized as very strong antioxidants and Ethyl acetate fraction is categorized as a strong antioxidant. The cytotoxic test results using crude extract obtained an LC50 value of 54,827 ppm which showed that Sikkam bark extract was toxic so that it was potential as an anticancer agent.

scholarly journals Antioxidant Potential of Crude Methanol Leaf extract and Fraction of Mallotus oppositifolius

2021 ◽  
pp. 168-184
Author(s):  
Ifeanyi Peter Onyeka ◽  
Felix Ahamefule Onyegbule ◽  
Christopher Obodike Ezugwu ◽  
Chibueze Jeremiah Ike ◽  
Chidalu B. Ikeotuonye
Keyword(s):  
Methyl Ester ◽  
Ethyl Acetate ◽  
Crude Extract ◽  
Leaf Extract ◽  
Antioxidant Activities ◽  
Radical Scavenging ◽  
Acid Methyl Ester ◽  
Ethyl Acetate Fraction ◽  
Antioxidant Potential ◽  
Acid Methyl

Mallotus oppositifolius (Geiseler) Mull. Arg.  (Euphorbiaceae) is a predominant edible shrub in Nigeria. It is locally used for treatment of common infections and wounds. This study evaluated the toxicity and antioxidant potential of the crude extract and fractions of M. oppositifolius using invitro 1,1-diphenyl-2-picrylhydrazine (DPPH) radical scavenging assay. The result showed abundance of flavonoids (28.21), tannins (17.72), alkaloids (7.79) and Saponins (0.23) in the crude extract of M. oppositifolius leaf extract. Also, the result showed that the LD50 is above 5000 mg/kg, therefore the crude extract is safe for consumption. The result further showed that the ethyl-acetate and butanol had significant (p<0.05) antioxidant activity when compared with the control at 125, 62.5 and 31.125 µg/ml respectively while at 250 and 500 µg/ml there was no significant different in the level of reduction of oxidation between the butanol and ethyl acetate when compared with the control. The result further showed that the butanol and ethyl-acetate fraction of M. oppositifolius had the best antioxidant potential among the fractions studied. The result of GCMS studies of ethyl-acetate fraction showed the presence of 7,10,13-eicosatrienoic acid methyl ester, korormicin, agosterol, phyllanthin, ricinoic acid, 2,4,6-decatrienoic acid 1a,2,5,5a,6,9,10,10a-octahydro-5,5a-dihydroxy-4-(hydroxymethyl)-1,1,7,9, limonin, com-pounds that have shown to possess strong antioxidant activities. Also, the GCMS result of butanol fraction showed the presence of seven compounds which include: ferulylhistamine, hippeastrine, 10,13-eicosadienoic acid methyl ester, ethyl 5,8,11,14-eicosatetraenoate, 6-hydroxy-powelline-N-nitroso-7-demethoxy-aldehyde, 6,7-epoxypregn-4-ene-9,11,18-triol-3,20-dione 11,18-diacetate and 5Alpha-androstane-3,17-dione 17-monooxime,  compounds which may serve as a new lead for the treatment of diseases associated with the oxidative stress.

Recombinant human granulocyte colony-stimulating factor: effects on normal and leukemic myeloid cells

Science ◽  
1986 ◽  
Vol 232 (4746) ◽  
pp. 61-65 ◽  
Author(s):  
LM Souza ◽  
TC Boone ◽  
J Gabrilove ◽  
PH Lai ◽  
KM Zsebo ◽  
...  
Keyword(s):  
Cell Line ◽  
Biological Activities ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Granulocyte Colony Stimulating Factor ◽  
Newly Diagnosed ◽  
Colony Stimulating Factor ◽  
Binding Studies ◽  
D Cells ◽  
Stimulating Factor

Experiments were conducted to isolate and characterize the gene and gene product of a human hematopoietic colony-stimulating factor with pluripotent biological activities. This factor has the ability to induce differentiation of a murine myelomonocytic leukemia cell line WEHI-3B(D+) and cells from patients with newly diagnosed acute nonlymphocytic leukemia (ANLL). A complementary DNA copy of the gene encoding a pluripotent human granulocyte colony-stimulating factor (hG-CSF) was cloned and expressed in Escherichia coli. The recombinant form of hG-CSF is capable of supporting neutrophil proliferation in a CFU-GM assay. In addition, recombinant hG-CSF can support early erythroid colonies and mixed colony formation. Competitive binding studies done with 125I-labeled hG-CSF and cell samples from two patients with newly diagnosed human leukemias as well as WEHI-3B(D+) cells showed that one of the human leukemias (ANLL, classified as M4) and the WEHI-3B(D+) cells have receptors for hG-CSF. Furthermore, the murine WEHI-3B(D+) cells and human leukemic cells classified as M2, M3, and M4 were induced by recombinant hG-CSF to undergo terminal differentiation to macrophages and granulocytes. The secreted form of the protein produced by the bladder carcinoma cell line 5637 was found to be O-glycosylated and to have a molecular weight of 19,600.

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