Development, preparation, and evaluation of a novel non-adjuvanted polyvalent dermatophytes vaccine.

Ringworm is a worldwide distributed contagious disease infecting both man and animals that constitute an economic, zoonotic, and health problem concern all over the world. During the last decade, attention has been directed to vaccination as an ideal approach to the control of such diseases. In the present study, non-adjuvanted polyvalent vaccines were prepared from locally isolated hot and virulent dermatophyte species, namely Trichophyton verrucosum (T. verrucosum), Trichophyton mentagrophytes (T. mentagrophytes), and Microsporum canis (M. canis) were immunologically evaluated. The prepared vaccine evaluation was focused on the aspects of immunogenicity and protective efficacy using guinea pigs. Both in its living or inactivated forms, the vaccine-induced significant humoral and cell-mediated immune responses and achieve proper protection of guinea pigs against challenging infections with homologous and heterologous dermatophyte strains. On the other hand, investigations on dermatophyte exo-keratinases showed that it was better produced and more expressed in a mineral-based medium containing pure keratin (3 g/L) than in the same medium with human hair supplementation (2.6 g/L). The maximum dermatophyte productivity of exo-keratinases was found to be between 18 and 21 days post-incubation. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two fractions with molecular weights of 40 kDa (fraction I) and 28 kDa (fraction II) have been identified in the culture filtrate of the three involved dermatophyte species. Both fractions demonstrated keratinolytic activity. The specific activity of the isolated keratinases (number of Keratinase units (KU)/mg protein) was stronger in fraction I, where it reached 18.75, 15.38, and 14 KU/mg protein as compared to 12.9, 8.74, and 12 KU/mg protein in fraction II of T. verrucosum, T. mentagrophytes, and M. canis, respectively. The dermatophyte exo-keratinases proved to be immunogenic as they stimulated high keratinase-specific antibody titers and induced strong delayed skin hypersensitivity reactions in vaccinated animals. Anti-keratinase-specific IgG was detected in sera of guinea pigs immunized with the inactivated or living polyvalent dermatophyte vaccines by a homemade enzyme-linked immunosorbent assay (ELISA) using dermatophyte exo-keratinases as coating antigen. The intradermal injection of dermatophyte exo-keratinases induced specific delayed skin reactions in guinea pigs immunized with the inactivated or the living polyvalent dermatophyte vaccines. The intradermal injection of dermatophyte exo-keratinases in the control non-sensitized guinea pigs was associated with itching, swelling, and bloody scar formation, however, no skin indurations were formed. The development of those post-exo-keratinases injection reactions in the control non-sensitized apparently healthy guinea pigs group, suggests an exo-keratinases possible role in the pathogenesis of dermatophytosis.

Sofosbuvir plus Daclatasvir with or without ribavirin for treatment of chronic HCV genotype 4 patients: real-life experience.

INTRODUCTION: New regimens involving direct-acting antiviral agents have recently been approved for the treatment of HCV. Our aim was to assess the efficacy and safety of 12 or 24 weeks of Sofosbuvir 400 mg plus Daclatasvir 60 mg, with or without ribavirin (800-1000 mg) in treating chronic hepatitis C genotype 4 patients. METHODS: This is an open-label observational study that describes the effect of 12 week or 24 weeks of daily oral Sofosbuvir (SOF) 400 mg plus Daclatasvir (DCV) 60 mg with or without ribavirin (RBV) with dose adjustment if indicated. It included the first 1168 patients that fulfilled the inclusion and exclusion criteria and treated in the Egyptian Liver Research Institute and Hospital, Mansoura, Egypt. RESULTS: Sustained viral response after 12 weeks of end of treatment (SVR12) was achieved in 96.6% (95% CI 95.1-98.2%) of the patients receiving 12 weeks of DCV + SOF treatment, in 95.7% (95% CI 93.6-97.8%) of the patients receiving 12 weeks of DCV + SOF + RBV, in 93.3% (95% CI 90.0-96.6%) of those receiving 24 weeks of DCV + SOF, and in 92.2% (95% CI 85.4-98.9%) of patients receiving 24 weeks of DCV + SOF + RBV treatment. SVR12 rate was significantly higher in patients with no cirrhosis receiving DCV + SOF only for 12 weeks or 24 weeks (97.4 and 97.4%, respectively) than in patients with cirrhosis (91.7 and 88.9%, respectively). The most common adverse events were fatigue, headache, insomnia, and anemia. No treatment-related serious adverse events or death were reported in the studied groups. CONCLUSION: Treatment with SOF (400 mg) plus DCV (60 mg), with or without RBV (800-1000 mg) for 12 or 24 weeks, was effective and well tolerated in chronic hepatitis C genotype 4 patients. SVR rates were higher for patients with no cirrhosis. Addition of RBV has benefit only in treatment-experienced group receiving 24 weeks.

Batch bioethanol production via the biological and chemical saccharification of some Egyptian marine macroalgae.

AIMS: Marine seaweeds (macroalgae) cause an eutrophication problem and affects the touristic activities. The success of the production of the third-generation bioethanol from marine macroalgae depends mainly on the development of an ecofriendly and eco-feasible pretreatment (i.e. hydrolysis) technique, a highly effective saccharification step and finally an efficient bioethanol fermentation step. Therefore, this study aimed to investigate the potentiality of different marine macroalgal strains, collected from Egyptian coasts, for bioethanol production via different saccharification processes. METHODS AND RESULTS: Different marine macroalgal strains, red Jania rubens, green Ulva lactuca and brown Sargassum latifolium, have been collected from Egyptian Mediterranean and Red Sea shores. Different hydrolysis processes were evaluated to maximize the extraction of fermentable sugars; thermochemical hydrolysis with diluted acids (HCl and H2 SO4 ) and base (NaOH), hydrothermal hydrolysis followed by saccharification with different fungal strains and finally, thermochemical hydrolysis with diluted HCl, followed by fungal saccharification. The hydrothermal hydrolysis of S. latifolium followed by biological saccharification using Trichoderma asperellum RM1 produced maximum total sugars of 510 mg g-1 macroalgal biomass. The integration of the hydrothermal and fungal hydrolyses of the macroalgal biomass with a separate batch fermentation of the produced sugars using two Saccharomyces cerevisiae strains, produced approximately 0·29 g bioethanol g-1 total reducing sugars. A simulated regression modelling for the batch bioethanol fermentation was also performed. CONCLUSIONS: This study supported the possibility of using seaweeds as a renewable source of bioethanol throughout a suggested integration of macroalgal biomass hydrothermal and fungal hydrolyses with a separate batch bioethanol fermentation process of the produced sugars. SIGNIFICANCE AND IMPACT OF THE STUDY: The usage of marine macroalgae (i.e. seaweeds) as feedstock for bioethanol; an alternative and/or complimentary to petro-fuel, would act as triple fact solution; bioremediation process for ecosystem, renewable energy source and economy savings.

Characteristics of systemic lupus erythematosus in a sample of the Egyptian population: a retrospective cohort of 1109 patients from a single center.

Introduction Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease that can vary among different ethnic and racial groups. Objective The objective of this paper is to study the prevalence of various manifestations of SLE in a sample of the Egyptian population. Patients and methods Information in this study was derived from the medical records of SLE patients who sought medical advice at a private clinic in Cairo from January 1980 to June 2016. Results This study included 1109 SLE patients, of whom 114 (10.3%) were males and 995 were females (89.7%). Mean age of onset was 25.89 ± 10.81 years, while the median of disease duration from the onset of the disease till the last recorded visit was 26 months. The most common cumulative manifestations were arthritis (76.7%), malar rash (48.5%), leukopenia (45.7%), and photosensitivity (45.6%). A total of 33.1% of the patients had nephritis, and neuropsychiatric lupus was present in 6.4% of the patients. Secondary antiphospholipid syndrome was present in 11.5% of the patients. Antinuclear antibody and anti-double-stranded deoxyribonucleic acid were present in 1060/1094 (96.9%) and 842/1062 (79.3%) of the patients, respectively. Antiphospholipid antibodies were present in 266/636 (41.8%) of the patients, anti-Smith in 54/240 (22.5%), anti-SSA/Ro in 61/229 (20.4%), and anti-SSB/La in 32/277 (11.6%) of the patients. Male patients had a statistically higher prevalence of nephritis ( p = 0.01), whereas arthritis and alopecia were statistically higher in females ( p = 0.012 and p = 0.006, respectively). Patients with juvenile onset had a statistically higher prevalence of nephritis and seizures ( p < 0.001 and p = 0.012, respectively). Conclusions Arthritis and malar rash represented the most common clinical manifestations. Male and juvenile-onset patients had a predilection toward a more severe disease. These results are in agreement with many studies conducted in the Middle East and worldwide. On the other hand, major organ involvement was exceptionally low, which is contradictory to several reports from the Middle East and across the globe.

A simple bedside blood test (Fibrofast; FIB-5) is superior to FIB-4 index for the differentiation between non-significant and significant fibrosis in patients with chronic hepatitis C.

INTRODUCTION: A simple non-invasive score (Fibrofast, FIB-5) was developed using five routine laboratory tests (ALT, AST, alkaline phosphatase, albumin and platelets count) for the detection of significant hepatic fibrosis in patients with chronic hepatitis C. The FIB-4 index is a non-invasive test for the assessment of liver fibrosis, and a score of ≤1.45 enables the correct identification of patients who have non-significant (F0-1) from significant fibrosis (F2-4), and could avoid liver biopsy. The aim of this study was to compare the performance characteristics of FIB-5 and FIB-4 to differentiate between non-significant and significant fibrosis. METHOD: A cross-sectional study included 604 chronic HCV patients. All liver biopsies were scored using the METAVIR system. Both FIB-5 and FIB-4 scores were measured and the performance characteristics were calculated using the ROC curve. RESULTS: The performance characteristics of FIB-5 at ≥7.5 and FIB-4 at ≤1.45 for the differentiation between non-significant fibrosis and significant fibrosis were: specificity 94.4%, PPV 85.7%, and specificity 54.9%, PPV 55.7% respectively. CONCLUSION: FIB-5 score at the new cutoff is superior to FIB-4 index for the differentiation between non-significant and significant fibrosis.

Resveratrol ameliorates oxidative stress and organ dysfunction in Schistosoma mansoni infected mice.

Schistosoma mansoni causes a major chronic debilitating disease in more than 230 million people around the world. The pathognomonic granuloma is a major cause of the oxidative stress encountered as a consequence of infection not only in the liver, but also in other important organs as spleen, lung, brain and kidney. Resveratrol administration at a dose of 20 mg/kg once daily for two weeks to mice infected with Schistosoma mansoni resulted in improvement in serum cholesterol and triglyceride levels. Enzymatic antioxidant profile showed significant modulations in Superoxide dismutase, catalase activities and reduced glutathione levels. Specific biomarkers for homeostasis of brain and lung i.e. Tau and RAGE respectively, showed significant improvement after resveratrol administration.

A novel nested multiplex PCR for the simultaneous detection and differentiation of Cryptosporidium spp., Enterocytozoon bieneusi and Encephalitozoon intestinalis.

Cryptosporidium spp. and microsporidia are opportunistic parasites affecting a wide range of hosts in which they can be potentially life threatening in immunocompromised individuals. Diagnosis usually relies on the identification of the stained Cryptosporidium oocyst or microsporidial spores, but these methods lack sensitivity and require highly trained technicians to perform and interpret the results. Molecular diagnosis offers an alternative with both superior sensitivity and specificity as compared to microscopy. Although replacing microscopy with nucleic acid based methods is hampered by the higher costs, in particular in developing countries, multiplexing the detection of more than one parasite in a single test has been found to be very effective and would decrease the cost of the test without the need for new equipment, as it would be the case for quantitative PCR. The method shown in this report for the simultaneous detection of Cryptosporidium spp., Enterocytozoon bieneusi and Encephalitozoon intestinalis by multiplex nested PCR, has proved to have several advantages versus microscopy such as higher sensitivity and specificity, low subjectivity and a minimal need for specialist's training to interpret the results. The present multiplex assay can fill an important gap to identify other possible causative agents of several diarrheal diseases which until present remain undiagnosed and can improve the epidemiology of the disease with a more reliable detection method.

Identification of a novel Assemblage B subgenotype and a zoonotic Assemblage C in human isolates of Giardia intestinalis in Egypt.

Giardia intestinalis (G. intestinalis) is a flagellate parasite which has been considered the most common protozoan infecting human. Molecular techniques are of great value in studying the taxonomy, the zoonotic potential of animal isolates and the correlation between the genetic variability of the parasite and the range of clinical symptoms observed in humans. The present work aims at genotyping G. intestinalis isolates from Egypt using molecular techniques. PCR targeting the ß-giardin locus, RFLP and sequencing were applied to 12 microscopically positive and 3 microscopically negative samples (which were positive by real time PCR targeting SSUr DNA). Two other loci, triose phosphate isomerase (TPI) gene and glutamate dehydrogenase (GDH) gene PCR and RFLP were also applied to all study isolates. The most frequent genotype was Assemblage B (13 out of 15), while Assemblage A and C were present in one sample each. This is the first report on zoonotic transmission of Assemblage C (dog genotype) to human in Egypt. Sequencing of the Assemblage B isolates revealed new subgenotypes with consistent mutations at specific positions, some of which were not characterized previously. The results shed light on the possibility that G. intestinalis can infect humans through a zoonotic route and open the door to wider investigations using different genetic loci to genotype Giardia isolates.

Molecular and epidemiological analysis of methicillin-resistant Staphylococcus aureus otorrhoea: hospital- or community-acquired?

OBJECTIVES: (1) To identify newly diagnosed cases of methicillin-resistant Staphylococcus aureus ear infection in our local population; (2) to determine the risk factors involved in these patients' clinical courses, and (3) to type the bacterial strains isolated and thus identify whether they were hospital- or community-acquired. DESIGN AND SETTING: Retrospective review of case notes, together with laboratory-based molecular studies in the departments of otolaryngology and medical microbiology in a university teaching hospital in Scotland, UK. SUBJECTS: Over a two-year period, 35 patients were identified with ear swabs positive for methicillin-resistant Staphylococcus aureus infection. These cases came from both hospital and community settings. MAIN OUTCOME MEASURES: (1) Identification of primary methicillin-resistant Staphylococcus aureus otorrhoea in patients with no previously documented colonisation; and (2) molecular typing of the strains isolated, using spa technology, to identify whether they were hospital- or community-acquired. RESULTS: Of the 35 positive patients, 27 were previously known carriers of methicillin-resistant Staphylococcus aureus. The eight patients with newly diagnosed methicillin-resistant Staphylococcus aureus otorrhoea presented initially in the community. All of these patients had had contact with hospital staff (as in-patients or out-patients) in the weeks preceding development of their ear infection. Using the spa technique for molecular typing, we identified hospital-acquired ('epidemic') methicillin-resistant Staphylococcus aureus type 15 in all eight patients' isolates. All were sensitive to topical gentamicin. CONCLUSIONS: In our cohort, hospital-acquired methicillin-resistant Staphylococcus aureus type 15 was the commonest cause of methicillin-resistant Staphylococcus aureus otorrhoea, despite the fact that these patients all first presented in the community. We believe that contact with hospital staff or health care workers is a risk factor for acquiring methicillin-resistant Staphylococcus aureus otorrhoea in the community.

Synthesis of novel pyrimidine-2,4-diones and 2-thioxo-pyrimidine-4-ones as potential anticancer and antimicrobial agents.

Two novel series of pyrimidine derivatives have been prepared, namely; 3,6-disubstituted perhydropyrimidine-2,4-diones 8a-l as well as 3,6-disubstituted 2-thioxo-perhydropyrimidine-4-ones 9a-k. The anticancer as well antimicrobial activities of these compounds and their open-chain counterparts have been determined.

Pyrimidines. Part II: Synthesis of novel pyrimidines, 1,2,4-triazolo[4,3-a]pyrimidin-7-ones and pyrimidino[2,1-c][1,2,4]triazin-8-ones for their antimicrobial and anticancer activities.

Five main classes of novel pyrimidine derivatives have been synthesized; namely 6-substituted phenyl-5-cyano-3-methyl-2-phenacylhydrazino-3,4-dihydropyrimidin-4-ones 4a-e; 6-substituted phenyl-2-arylidene hydra-zino-5-cyano-3-methyl-3,4-dihydropyrimidin-4-ones 5a-i; 6-substituted phenyl-2-acylhydrazino-5-cyano-3-methyl-3,4-dihydropyrimidin-4-ones 7a-d, 8a-e and 9a-c; three novel series of 1,2,4-triazolo[4,3-a] pyrimidones 10a,b, 11a-d and 12a-d and 6-substituted phenyl-7-cyano-9-methyl-3-phenyl or 4-chlorophenyl-4,9-dihydropyrimido[2,1-c][1,2,4] triazin-8-ones 13a-c. Besides, the azide compound 2-azido-5-cyano-3-methyl-6-phenyl-3,4-dihydropyrimidin-4-one 6 was also synthesized. The prepared compounds were tested for antimicrobial and anticancer activity. Compounds 4b and 4d showed promising activity against Escherichia coli. Compounds 3c, 5c, 5e, 5g and 7b were active in the three cell line antitumor one dose primary assay and were evaluated in the 60 human tumor full panel cell line invitro screening. Compound 5c showed promising activity against all types of leukemia especially leukemia K-562 and leukemia SR with GI50 = 1.61 and 2.63 mmol/l respectively.

Thioglycolic acid and pyrazole derivatives of 4(3H)-quinazolinone: synthesis and antimicrobial evaluation.

New derivatives of 4(3H)-quinazolinone were synthesized and evaluated for their antibacterial and antifungal activity. The derivatives are: 3-Aryl-2-[3-aryl-1-(carboxymethylthio)-3-oxopropyl)]-4(3H)-quinazolinones 2a-f; 3-Aryl-2-(3-aryl-1H-pyrazol-5-yl)-4(3H)-quinazolinones 3a-f; 3-Aryl-2-(1,3-diaryl-1H-pyrazol-5-yl)-4(3H)-quinazolinones 4a-f; 3-Aryl-2-(3-aryl-1-thiocarbamoyl-1H-pyrazol-5-yl)-4(3H)-quinazolinones 5a-f; 3-Aryl-2-[3-aryl-1-(carboxymethylthio)-3-hydroxyiminopropyl)]-4(3H)- quinazolinones 6a-f; and 3-Aryl-2-[3-aryl-1-(carboxymethy- lthio)-3-thiocarbamoyliminopropyl)]-4(3H)-quinazolinones 7a-f. Some of the tested compounds showed activity comparable to that of the standard references used.

Synthesis and antimicrobial activity of novel pyrazole, pyrazoline, pyrazolinone and pyrazolidinedione derivatives of benzimidazole.

Four novel series of pyrazolylbenzimidazole derivatives have been prepared, namely 2-[(1-substituted phenyl-3,5-dimethyl-4-pyrazolyl)methyl]benzimidazole 5a-d 2-[(1-substituted phenyl-3-methyl-5-oxo-4,5-dihydro-4-pyrazolyl-4-yl)methyl]benzimidazoles 6a-d; 2-[(1-substituted phenyl-3,5-dioxopyrazolidin-4-yl)methyl]benzimidazoles 7a-d and 2-[(4-(1-phenyl-5-aryl-4,5-dihydro-3-pyrazolyl)phenylaminoacetyl]thio- methyl)-benzimidazoles 12a-e. The antimicrobial testing of the prepared compounds was performed using Escherichia Coli (NCTC 5933) as Gram-negative bacteria, Staphylococcus aureus (NCTC 4163) as gram-positive bacteria and Candida albicans (NCTC 5310) as yeast like fungi. The most potent compound was the pyrazolone 6a which exhibits interesting antibacterial activity against the gram-negative bacteria E. coli.

Synthesis and antimicrobial testing of 4H-1,2,4-triazole, 1,2,4-triazolo[3,4-b][1,3,4]thiadiazole and 1,2,4-triazolo[3,4-b][1,3,4]thiadiazine derivatives of 1H-benzimidazole.

Three novel series of benzimidazole derivatives namely 6-substituted 3-[1-(2-alkyl-1 H-benzimidazolyl)methyl]-1,2,4-triazolo[3,4-b][1,3,4] thiadiazoles 5a-h, 6-substituted 3-[1-(2-alkyl-1H-benzimidazolyl)methyl]-7H-1,2,4 -triazolo[3,4-b]-[1,3,4]thiadiazines 6a-j and 6-thioxo-3-[1-(2-alkyl-1H-benzimidazolyl)methyl]-5,6-dihydro-1,2,4 -triazolo[3,4-b][1,3,4]-thiadiazoles 7a, b have been prepared by cyclization of the key intermediate 1-[(4-amino-5-mercapto-4 H-1,2,4-triazol-3-yl)methyl]-2-alkyl-1 H-benzimidazoles 3a, b. Furthermore, 1-[(4-arylideneamino-5-mercapto- 4H-1,2,4-triazol-3-yl)-methyl]-2-alkyl- 1 H-benzimidazoles 4a-h have been prepared and some of them were cyclized to 6-substituted 3-[1-(2-alkyl-1H-benzimidazolyl)methyl]-1,2,4-triazolo [3,4-b][1,3,4]thiadiazoles 5d, h using thionyl chloride. The prepared compounds were tested for antimicrobial activity in vitro; they showed moderate activity.

Synthesis and antimicrobial testing of novel oxadiazolylbenzimidazole derivatives.

Three novel series of oxadiazolylbenzimidazole derivatives have been prepared, namely 1-[(2-alkylthio or aralkylthio-1,3,4-oxadiazol-5-yl)methyl]-2-alkyl-1 H-benzimidazoles 3a-f; 1-[(3-substituted aminomethyl-2-thioxo-2,3-dihydro-1,3,4-oxadiazol-5-yl)methyl ]-2-alkyl-1 H-benzimidazoles 4a-f and 1-[(2-substituted amino-1,3,4-oxidiazol-5-yl)methyl]-2-alkyl-1 H-benzimidazoles 6a-j. The antimicrobial testing of the prepared compounds was performed, some of them showed week activity.

Ultrastructure of resting and rh-GMCSF-treated human macrophages derived from blood monocytes.

The ultrastructure of cultured blood monocyte-derived human macrophages was investigated and correlated under the effect of different doses of rh-GMCSF (dose 1 = 25 IU/ml, dose 2 = 125 IU/ml and dose 3 = 250 IU/ml). Resting macrophages showed irregular cell borders and pseudopodia pushed out in all directions. Their cytoplasm depicted rough endoplasmic reticulum and Golgi complex in the perinuclear area. Lipid globules, primary lysosomes and mitochondria were characteristically prominent. rh-GMCSF-stimulated macrophages were more voluminous and their nuclei were irregular in outline, with predominance of euochromatin over heterochromatin. The cytoplasm was overcrowded by an increasing number of organelles including lysosomes, phagolysosomes and mitochondria. Golgi complex demonstrated a wide-spread distribution along the cells, with profound membrane expansion and cisternal dilatation; especially, in cells treated with dose 2. Electron dense osmiophilic deposits (collapsed membranes) were seen in association with lipid globules, which were commonly polarized at cell peripheries. Most of these changes were dose dependent. However, cells treated with dose 3 manifested additionally well-developed centrioles, inapparent nuclear membrane, display of microfilaments and well-established adhesions. The demonstrated ultrastructural changes in rh-GMCSF-treated human macrophages indicated pronounced activation, which supports the reported clinical effect of this cytokine.

Enhancement of leishmanicidal activity of human macrophages against Leishmania major and Leishmania donovani infection using recombinant human granulocyte macrophage colony stimulating factor.

The in vitro effect of recombinant human Granulocyte Macrophage Colony Stimulating Factor (rh-GMCSF) on the leishmanicidal activity and superoxide anion productivity of macrophages derived from human blood monocytes (MOs) were investigated. MOs treated with 25, 125, or 250 U/mL of rh-GMCSF for 72 h prior to infection with leishmania parasites, manifested significant dose-dependent increase in its leishmanicidal activities against Leishmania major and Leishmania donovani parasites. The percentage of increase in leishmanicidal activity of L. major-infected MOs were 22.71, 64.34 and 81.34, respectively while in L. donovani-infected MOs, it reached 3.01, 32.28 and 74.38, respectively. Treatment of leishmania-infected MOs with rh-GMCSF (250 U/mL) for different periods of time up to 96 hours, induced a significant time-dependent reduction in the percentage of infected cells and the parasitic load (No. of amastigotes/100 MOs). After 96 h of treatment with rh-GMCSF, the percentages of reduction in the infection rates were 82.45 in L.major-infected MOs (p < 0.001) and 39.65 in L. donovani-infected cells (p < 0.01). The percentage of reduction in the parasitic load reached 90.82 (p < 0.001) and 36.6 (p < 0.05) in MOs infected with L. major and L. donovani, respectively. The priming effect of rh-GMCSF on superoxide anion production by human MOs stimulated with phorbol myristate acetate (PMA) was both dose-dependent and time-dependent. In 72 hour-old human MOs, the maximum superoxide anion release was generated by MOs primed for 45 min with 500 U/mL of rh-GMCSF. These cells produced 8.960 +/- 2.075 nmol/5 x 10(4) MOs/ 180 min as compared to 4.563 +/- 1.773 nmol/5 x 10(4) unprimed cell control/180 min (p < 0.001).

Effect of rh-GMCSF and rh-GCSF on oxygen free radical production by human neutrophils and blood monocyte-derived human macrophages.

The in vitro effect of recombinant human granulocyte-macrophage colony stimulating factor (rh-GMCSF) and recombinant human granulocyte colony stimulating factor (rh-GCSF) on oxygen free radical (OFR) generation by human neutrophils and blood monocytes derived human macrophages stimulated with phorbol myristate acetate was investigated and compared. The production of OFR by neutrophils and macrophages was time dependent, and the maximum release of OFR by neutrophils and macrophages was measured 90 and 180 min after stimulation with phorbol myristate acetate, respectively. The priming effects or rh-GMCSF and rh-GCSF on OFR production by human neutrophils and macrophages was dose dependent. The maximum generation of OFR by neutrophils occurred when primed with 1,000 U/ml of rh-GMCSF and reached 2.383 +/- 0.191 nmol/10(5) neutrophils/90 min as compared with 1.072 +/- 0.113 nmol/10(5) neutrophils/90 min in the unprimed controls. This represents a 122.20% increase in OFR generation (p < 0.001). However, the percentage of maximum increase in OFR production was 57.84 when neutrophils were primed with a concentration of 5,000 U of rh-GCSF/ml. In 72-hour-old human macrophages, much higher levels of OFR production as compared with neutrophils were measured following stimulation with phorbol myristate acetate. The maximum generation of OFR was measured in macrophages primed for 45 min with 500 U/ml of rh-GMCSF. These cells produced 8.960 +/- 2.075 nmol/5 x 10(4) macrophage/180 min as compared with 4.563 +/- 1.773 nmol/5 x 10(4) unprimed macrophages/180 min (p < 0.001). In macrophages primed with rh-GCSF, however, the maximum OFR production was induced by a dose of 5,000 U/ml and reached 6.902 +/- 1.463 nmol/5 x 10(4) macrophages/180 min as compared with 4.563 +/- 1.773 nmol/5 x 10(4) macrophages/180 min in the unprimed controls (p < 0.05). In conclusion, the priming effect of rh-GMCSF on OFR generation by human macrophages and neutrophils was more potent than that of rh-GCSF, both in the extent of augmentation and in the dose required to produce maximum OFR generation.