Determination of Alternate Pathway Complement Kinetics by Electron Spin Resonance Spectroscopy

Size: px
Start display at page:

Download "Determination of Alternate Pathway Complement Kinetics by Electron Spin Resonance Spectroscopy"

Transcription

1 Determination of Alternate Pathway Complement Kinetics by lectron Spin Resonance Spectroscopy DAN A. HAWLY, M.D., FRDRICK W. KLINHANS, PH.D., AND JAMS L. BISCKR, M.D., PH.D. We describe a technic that measures the kinetics of the metabolic burst of peripheral blood neutrophils (PMN) by electron spin resonance (SR) spin trapping. Using this technic, a functional assay for the kinetics of the alternate pathway complement cascade is developed. PMN were stimulated by phagocytosis of opsonized zymosan (OpZym) to undergo the metabolic burst and the resulting free hydroxyl radicals generated during the burst were trapped using 5,5-dimethyl-l-pyrroline- N-oxide (DMPO) to form a stable spin adduct. Rapid sequential measurements of the spin adduct concentration were made using a computer-controlled SR spectrometer to follow the time course of the metabolic burst. Differences in the results obtained with OpZym and with PMN incubated with unopsonized zymosan (Zym) and serum yields information about the kinetics of opsonization. The kinetics of the alternate complement pathway opsonization in sera from normal and C8-deficient patients were compared and found not to be affected significantly by the absence of C8 complement protein. (Key words: Complement kinetics; lectron spin resonance; SR; Spin trap; DMPO; PMN) Am J Clin Pathol 1983; 79: TH PROTINS of the alternate complement pathway have been elucidated, but the kinetics of the cascade are not well-characterized During activation of the alternate pathway, opsonins are generated that are capable of coating particles in the surrounding microenvironment. In the presence of neutrophils, the opsonized particles may be phagocytized, and the metabolic burst within the neutrophils occurs. The purpose of this study is to demonstrate a functional assay for the alternate complement pathway and for the events involved in the metabolic burst of peripheral blood neutrophils (PMN). The alternate pathway can be activated using unopsonized zymosan (Zym), an extract of yeast cell wall prepared from Saccharomyces cerevisiae UM PMN phagocytize the opsonized zymosan (OpZym), the metabolic burst occurs, and free hydroxyl radicals (OH") are produced. 9 The feasibility of trapping this OH* using 5,5- dimethyl-1-pyrroline-n-oxide (DMPO) and detecting the resulting free radical (DMPO/OH)' using electron spin resonance (SR) spectroscopy recently has been demonstrated ' 5 We have developed a computer- Received August 13, 1982; accepted for publication September 9, Address reprint requests to Dr. Hawley: Department of Medical Research, Methodist Hospital of Indiana. Inc North Capitol Avenue, Indianapolis, Indiana Departments of Pathology and Medical Research, Methodist Hospital Graduate Medical Center and Department of Physics, Indiana University-Purdue University at Indianapolis, Indianapolis, Indiana automated SR method that rapidly and sequentially measures the amplitude of the spin adduct signal during the metabolic burst. The kinetics of the alternate pathway complement cascade were measured using an in vitro system containing PMN, DMPO, and either OpZym or Zym and serum. The time delay occurring when Zym and serum are used in place of OpZym reflects the length of time required for opsonization of the Zym by the serum. This opsonization time was determined for normal serum and for serum from a patient with a deficiency of the eighth component of complement. The influence of various concentrations of complement was investigated by measuring the metabolic activity of PMN activated by Zym in serial dilutions of AB serum. The effect of temperature on opsonization, phagocytosis, and PMN metabolism also was investigated. To confirm that OH* was the free radical trapped, inhibitors of the metabolism of PMN were added to the reaction mixture. 515 The inhibitors included superoxide dismutase (SOD), catalase, and nitroblue tetrazolium (NBT). Materials and Methods Whole blood with 100 U/mL sodium heparin (Panheprin, Abbott) was collected from normal, healthy adults who gave informed consent for venepuncture. PMN from whole blood were isolated by density gradient centrifugation with Ficoll-Hypaque and hypotonic lysis of erythrocytes. PMN were resuspended at 4 X 10 7 PMN/mL in modified Krebs-Ringer's phosphate glucose buffer (KRP). 7 Zym (Sigma, St. Louis, MO) was suspended at 8.28 mg/ml in KRP. OpZym was prepared by incubating the Zym suspension in 14% pooled human AB serum for 10 minutes at 37 C. For several experiments, Zym was suspended in KRP containing SOD (Sigma) at 80 Mg/mL, catalase (Sigma) at 960 fig/ m L, or NBT (Sigma) at 165 Mg/mL. A 1.0 molar aqueous /83/0600/0673 $01.05 American Society of Clinical Pathologists 673

2 674 HAWLY, KLINHANS, AND BISCKR A.J.C.P. June 1983 FIG. 1. SR spectrum of (DMPO/OH) - adduct produced by stimulated PMN. Signal obtained after 12 minutes incubation at 37 C of 50 yjl of PMN in KRP (4 X 7/mL). 50 nl zymosan (8.28 mg/ml). 25 nl aqueous DMPO (1 M). and 25 ML AB serum. Spectrometer Settings: Field 3313 G, Freq 9.3 GHz. power 60 mw (dual cavity), modulation khz. time constant 0.25 seconds, gain 2.5 X 10 4, computer controlled sweep 4 minutes total. solution of DMPO (Aldrich, Milwaukee, WI) was used for spin trapping. Control serum was obtained by pooling normal human AB sera. C8-deficient serum was obtained from a patient with known C8 deficiency demonstrated by absence of C8 on immunoelectrophoresis and absence of detectable hemolytic complement activity. Sera were frozen at -70 C and, when necessary, heat-inactivated at 56 C for 30 minutes. SR samples were prepared by combining 50 nl of PMN, 50 ixl of the suspension of Zym, 25 pl of KRP buffer, 25 fil of the DMPO solution, and 25 ixh of serum. For some experiments OpZym was substituted by Zym and buffer was replaced with serum, and for some experiments the serum was heat-inactivated. For all assays the suspensions were mixed quickly in a glass Table I. SR Determined'PMN Metabolic Activity Using Opsonized Zymosan and Zymosan Plus Serum* Time of Peakjij (minutes) Opsonized zymosanf 3.7 ± 0.4 (N) (7) Zymosan plus serumf 6.3 ± 0.4 (N) (7) Peak Amplitude (mm) 247 ± 7 (7) 208 ± 14 (7) test tube and drawn into a 100-/iL capillary tube (Clay Adams, 4625). The capillary tubes were sealed with Critoseal (Sherwood, St. Louis, MO) and inserted into the spectrometer cavity. The temperature of the cavity was maintained at 37 ± 0.5 C. We performed paired measurements comparing the metabolism of PMN phagocytizing OpZym with the metabolism of PMN phagocytizing Zym in the presence of serum. Similarly, we compared the metabolism of PMN that phagocytized Zym while incubated in pooled AB serum with the metabolism of PMN phagocytizing Zym while incubated in C8-deficient serum. The statistical significance of the results were evaluated with the paired Mest. Spectra were obtained using a Varian l09 X band spectrometer with an dual sample cavity. Spectra of the second line of the spin adduct quartet were obtained using settings of: frequency = 9.17 GHz, field = 3254 gauss centered on the second line in the spectrum, sweep width = 3 gauss, sweep time = 60 seconds, microwave power = 60 mw (into dual cavity), modulation amplitude = 2 gauss at 100 khz, gain = 2.5 X 10 4, and detection time constant of 1 second. Some spectra were obtained by using a 90-second sweep with a 4- second time constant. These fast sweep rates and long time constants caused some spectral distortion, but they provided maximum sensitivity and no loss of internal consistency. The spin adduct SR signal decays in a period of * The reaction mixtures contained 5011LPMN in KRP (4 X I0 7 KRP (8.28 mg/ml) and 50 ML KRP; (U) 50 ML zymosan. 25 nl KRP. PMN/mL). 25 nl aqueous and 25 nl AB serum. The ordinate is the (DMPO/OH)' signal amplitude normalized to a spectrometer gain of 2.5 X Spectrometer DMPO (I'M), and 25 j*l KRP plus 25 nl opsonized zymosan (8.28 mg/ml) or 25 tilab serum plus 25 fil zymosan. t Mean ± standard deviation. Number of measurements = 7. settings: H 0 = 3254 G, f = 9.17 GHz, AH = 3 G. P = 60 mw (dual X Measured from addition of serum or opsonized zymosan. cavity), t = 90 seconds, modulation = 2 G at 100 khz. and time The peak time differences are significant at the P = level. constant = 4 seconds. d) "0 0 +) Q- < ca s in T i me (m i n) FIG. 2. PMN metabolic burst vs. time, stimulated with opsonized zymosan (O) and with zymosan plus serum (U). The reaction mixtures contained 50 ML PMN in KRP (4 X 10 7 PMN/mL), 25 ML aqueous DMPO (1 M), and the following: (O) 50 nl opsonized zymosan in ca (M

3 Vol. 79 No. 6 COMPLMNT KINTICS BY SR 675 minutes. 13 In an experiment lasting 20 minutes or longer, the signal at any given time does not represent the integrated spin adduct production from time zero. Rather, it represents the average of spin adduct concentration over the previous few minutes with the spin adduct concentration from more recent times weighed more heavily. Consequently, the underlying process of OH* production is observed with a time resolution of only a few minutes, despite the somewhat faster sampling rate of the experimental procedure. The spectrometer was controlled by a microcomputer (Hewlett-Packard 9825T ) programmed to sweep through the second spectral line once every 60 or 90 seconds for a total period of 20 to 60 minutes. Spectra were stored on a floppy disk for subsequent analysis and graphing of spectral line amplitude versus time. A computer-controlled plotter (Hewlett-Packard 9872B) generated graphs directly from the data files. Results A typical (DMPO/OH)* spin adduct spectrum obtained with stimulated PMN is shown in Figure 1. The four-line spectrum had hyperfine splitting constants of A N = A H = 14.8 gauss and a 1:2:2:1 intensity distribution, which is consistent with the reported SR parameters of the (DMPO/OH)* adduct Further experiments were conducted by measuring only the second line of this signal and plotting its amplitude versus time. A comparison of the metabolic burst of normal PMN phagocytizing OpZym with the metabolic burst of normal PMN phagocytizing Zym plus serum is presented in Table 1, and typical results are shown in Figure 2. No signal was obtained using heat-inactivated AB serum instead of AB serum. The peak of metabolic burst activity from PMN phagocytizing OpZym occurred 2.6 minutes before the peak metabolic activity from PMN phagocytizing Zym in serum. The peak amplitude of 150 FIG. 3. ffect of inhibitors on stimulated PMN metabolic burst. Reaction mixtures consisted of 50 ML' PMN in KRP (4 X 10 7 PMN/ ml), 25 nl aqueous DMPO (1 M), 50 JIL AB serum and: (*) 50 ^L zymosan in KRP (8.28 mg/ml); (C) zymosan + catalase (960 jig/ml); (N) zymosan + NBT (165 Mg/mL); and (S) zymosan + SOD (80 \x%l ml). PMN phagocytizing OpZym was 19% higher than the peak amplitude of PMN phagocytizing Zym in serum (Table 1). C8-deficient serum used in place of normal pooled AB serum for opsonization of zymosan produced a metabolic peak 0.4 minutes later and the peak amplitude diminished by 12% (Table 2). Results of experiments utilizing SOD, catalase, or NBT in the incubation mixture are shown in Figure 3. Catalase reduced the amplitude of the signal by 34%, and SOD completely suppressed the signal. NBT decreased the signal amplitude by 40%, and a black precipitate was noted in the capillary tubes at the comple- 200 Table 2. SR Determined Metabolic Burst Activity Using Pooled AB Serum or C8-deficient Serum for Opsonization of Zymosan* Pooled ABf (N) C8-deficientt (N) Time of Peakt (minutes) 7.2 ± ± 0.4 Peak Amplitude (mm) 170 ± ± 10 FIG. 4. ffect of serum dilution on PMN metabolic burst. Reaction The reaction mixtures contained 50 nlpmn in KRP (4 X 10' PMN/mL). 25 /il aqueous mixtures consisted of 50 ML PMN in KRP (4 X 10' PMN/mL). 50 DMPO (1 M), 25ML zymosan in KRP (8.28 mg/ml). and 25 JIL serum. ML zymosan in KRP (8.28 mg/ml), 25 ML aqueous DMPO (1 M), t Mean ± standard deviation. Number of measurements = 5. and: (D) 50 nh AB serum; (*) 25 ML AB serum and 25 iil KRP; $ Measured from addition of serum. (A) 12.5 nl AB serum and 37.4 n\ KRP; (+) 6 ML AB serum and 44 Peak time and amplitude differences are significant at the P = 0.01 level. ML KRP.

4 676 HAWLY, KLINHANS, AND BISCKR A.J.C.P. June 1983 Table 3. Quantitative Levels of C3, C3-activator, and C4 in Pooled AB Serum and C8-deficient Serum Normal A3 Serum C8-deficient Serum C3* 111 mg/dl 190 mg/dl C3-activatorf 28.6 mg/dl 41 mg/dl C4* 20.8 mg/dl 27.7 mg/dl * Determined by radial immunodiffusion. t Determined by laser nephelometry. tion of the SR measurement. The reaction mixture with NBT was examjned by light microscopy and the black precipitate was consistent with intracellular formazan. 12 The metabolism of the PMN phagocytizing Zym or OpZym is related to the concentration of serum used for opsonization. This is shown in Figure 4. The serum concentration was varied from 28% to 3.5%. Peak response time changed from 5.6 minutes with 28% serum to 26.6 minutes with 3.5% serum. The peak amplitude dropped with both high (28%) and low (1%, not shown) serum concentrations, but it remained fairly constant over the range of 14% to 3.5% concentration. The amplitude of the signal with any one serum concentration was also dependent upon the net concentrations of zymosan and PMN, and the zymosan:pmn ratio (results hot shown). The serum concentration results may be interpreted to mean that complement concentration is an important factor in determining the peak time and amplitude of response. Therefore, complement levels were determined for the pooled AB and C8-deficient serum. Quantitative levels of C3, C3-activator (also known as B), and C4 were determined by radial immunodiffusion and/or laser nephelometry for both the patient and the pooled serum (Table 3). 2 C3, C3-activator, and C4 levejs averaged 49% higher in the C8-deficient serum than in the AB serum. To determine sensitivity to incubation temperature, SR measurements of the metabolism of PMN were made at several temperatures. Although the time of peak response was fairly constant between 35 C and 38 C, there was a doubling of the response time by 39 C. Above 37 C, the amplitude of the response diminished. Discussion The peak metabolic activity of PMN incubated with OpZym occurred after 3 to 5 minutes of incubation. We found that significant alternate pathway opsonization of zymosan occurred within 2 to 3 minutes. Studies conducted by other investigators using different antigens and methods have shown similar patterns of results. 1-3,8,11.16,18.19,20 Deficiency of C8 in serum did not have a significant effect on the kinetics of the opsonization of zymosan, and this is consistent with the known noninvolvement of C8 in the alternate pathway." 18 Inhibitors of OH" production, including SOD and catalase, caused decreased signal amplitudes. 4 ' 515 We found similar results with NBT. Formazan production and decreased signal amplitude were observed when NBT was present in the reaction mixture. The likely explanation is that NBT reacts with superoxide to produce formazan, and this reaction decreases the amount of superoxide available for OH' production. Physiologic temperatures were optimal for OH" production. Increased temperatures caused diminished OH' production and others have reported similar findings Serum dilution curves showed incremental increases in opsonization time that corresponded to decremental changes in serum concentration. These increases in opsonization time are likely a result of the decreased levels of complement. In summary, we have shown that the kinetics of the alternate pathway of complement can be determined using SR spin trapping technics. No appreciable differences are found between pooled AB serum and C8- deficient serum. Acknowledgment. The authors wish to thank Brian G. Davies and Steven T. Barefoot, DDS for assistance in acquiring the SR data. References 1. Allen RC: valuation of serum opsonic capacity by quantitating the initial chemiluminescent response from phagocytizing polymorphonuclear leukocytes. Infect Immun 1977; 15: Biesecker JL, Triplett DA, Miller GA, Hawley DA: The pathologist as a consultant in the laboratory evaluation of the patient with recurrent infections. American Society of Clinical Pathologists Workshop Manual. Chicago, American Society of Clinical Pathologists, 1979 ' 3. Cohen HJ, Newburger P, Chovaniec M, Whitin JC, Simons R: Opsonized zymosan-stimulated granulocytes activation and activity of the superoxide-generating system and membrane potential changes. Blood 1981; 58: Finkelstein, Rosen GM, Rauckman J, Paxton J: Spintrapping of superoxide. Mol Pharmacol l'979; 16: Green MR, Hill HAO, Okolow-Zubkowska MJ, Segal AW: The production of hydroxyl and superoxide radicals by stimulated human neutrophils measurements by PR spectroscopy. FBS Lett 1979; 100: Harbour JR, Chow V, Bolton JR: An electron spin resonance kudy of the spin adducts of OH and H0 2 radicals with nitrones in the ultraviolet photolysis of aqueous hydrogen peroxide solutions. Can J Chem 1974; 52: Hawley DA, MillerGA, Biesecker JL: Normal neutrophil function in human renal allograft recipients receiving Minnesota antiiymphoblast globulin (MAG). Am J Clin Pathol 1981; 76:29^ 33

5 Vol. 79 No. 6 COMPLMNT KINTICS BY SR Klebanoff SJ, Clark RA: Iodination by human polymorphonuclear leukocytes: a re-evaluation. J Lab Clin Med 1977; 89: Klebanoff S}, Clark RA: The neutrophil: function and clinical disorders, Chap 6; The metabolic burst, Chap 7, anti-microbial systems. Amsterdam, North-Holiand, 1978, pp Kleinhans FW, Hawley DA, Davies BG, Biesecker JL, Goodman MR: lectron paramagnetic resonance assay of serum from systemic lupus erythematosus patients. Bulletin of the American Physical Society 1981; 26: Muller-berhardt HJ: The complement system^ The plasma proteins, 2nd dition. dited by FW Putnam. New York, Academic Press, 1975, pp Nathan DG, Baehner RL, Weaver DK: Failure of nitroblue tetrazolium reduction in the phagocytic vacuoles of leukocytes in chronic granulomatous disease. J Clin Invest 1969; 48: Okolow-Zubkowska MJ, Hill HAO: Spin trapping of superoxide and hydroxyl radicals produced by stimulated human neutrophils. Developmental Biochemistry 1980; 11 B: Pillemer L, Blum L, Lepow IH, Ross OA, Todd W, Wardlow AC: The properdin system and immunity. Demonstration and isolation of a new serum protein, properdin, and its role in immune phenomena. Science 1954; 120: Rosen H, Klebanoff SJ: Hydroxyl radical generation by polymorphonuclear leukocytes measured by electron spin resonance spectroscopy. J Clin Invest 1979; 64: Rossi F, Romeo D, Patriarca P: Mechanism of phagocytosis-associated oxidative metabolism in polymorphonuclear leukocytes and macrophages. J Reticuloendothel Soc 1972; 12: Rustagi PK, LoBuglio AF: Phagocytosis of technetium-99m sulfur colloid by human polymorphonuclear leukocytes. J Immunol Methods 1978; 23: Schreiber RD, Morrison DC, Podack R, Muiler-berhard HJ: Bactericidal activity of the alternative complement pathway generated from 1 i isolated plasma proteins. J xp Med 1979; 149: Verhoef J, Peterson PK, Quie PG: Kinetics of staphylococcal opsonization, attachment, ingestion, and killing by human polymorphonuclear leukocytes: a quantitative assay using ('H) thymidine labeled bacteria. J Immunol Methods 1977; 14: Williams AJ, Hastings MJG, asmon CSF, Cole PJ: Factors affecting the in vitro assessment of opsonization: a study of the kinetics of opsonization using the technique of phagocytic chemiluminescence. Immunology 1980; 41: i

Supplementary Figure 1. DTPA does not interfere with the activity of catalase. Dependency of CAT activity on DTPA concentration at 25 C.

Supplementary Figure 1. DTPA does not interfere with the activity of catalase. Dependency of CAT activity on DTPA concentration at 25 C. Supplementary Figure 1. DTPA does not interfere with the activity of catalase. Dependency of CAT activity on DTPA concentration at 25 C. CAT (40 nm) was pre-incubated for 120 min with DTPA (0-10 mm) before

More information

Improved Immunodiagnosis of Neutrophil Dysfunction in the Newborn and Infant

Improved Immunodiagnosis of Neutrophil Dysfunction in the Newborn and Infant ANNALS OF CLINICAL AND LABORATORY SCIENCE, Vol. 12, No. 2 Copyright 1982, Institute for Clinical Science, Inc. Improved Immunodiagnosis of Neutrophil Dysfunction in the Newborn and Infant ALAN B. LOREN,

More information

CHAPTER 4 IMMUNOLOGICAL TECHNIQUES

CHAPTER 4 IMMUNOLOGICAL TECHNIQUES CHAPTER 4 IMMUNOLOGICAL TECHNIQUES Nitroblue Tetrazolium Chloride (NBT) Reduction test NBT reduction test was evaluated by employing the method described by Hudson and Hay,1989 based upon principle that

More information

Candidacidal Activity of Myeloperoxidase: Mechanisms of Inhibitory Influence of Soluble Cell Wall Mannan

Candidacidal Activity of Myeloperoxidase: Mechanisms of Inhibitory Influence of Soluble Cell Wall Mannan INFECTION AND IMMUNITY, OCt. 1983, p. 76-80 0019-9567/83/100076-05$02.00/0 Copyright 1983, American Society for Microbiology Vol. 42, No. 1 Candidacidal Activity of Myeloperoxidase: Mechanisms of Inhibitory

More information

Complement. Definition : series of heat-labile serum proteins. : serum and all tissue fluids except urine and CSF

Complement. Definition : series of heat-labile serum proteins. : serum and all tissue fluids except urine and CSF Complement Complement Definition : series of heat-labile serum proteins Site : serum and all tissue fluids except urine and CSF Synthesis : in liver appear in fetal circulation during 1 st 13 W Function

More information

Supplemental Materials

Supplemental Materials Supplemental Methods Spin trap and electron paramagnetic resonance spectroscopy Hypoxanthine (1 mmol=l) plus xanthine oxidase (.1 u=ml) were incubated in Krebs solution at 378C bubbled with 95% O 2 and

More information

Dependent Chemiluminescence Response and Bacterial Susceptibility to Phagocytosis

Dependent Chemiluminescence Response and Bacterial Susceptibility to Phagocytosis INFECTION AND IMMUNITY, Nov. 198, p. 37-374 19-9567/8/11-37/5$2./ Vol. 3, No. 2 Correlation Beteen Measurements of the Luminol- Dependent Chemiluminescence Response and Bacterial Susceptibility to Phagocytosis

More information

4 Development of an ESR online-method for the monitoring of in vitro fat digestion

4 Development of an ESR online-method for the monitoring of in vitro fat digestion 4 Development of an ESR online-method for the monitoring of in vitro fat digestion 4.1 Introduction When regarding the oral administration of lipid-based nanocapsules, gastrointestinal digestion will play

More information

The term complement refers to the ability of a system of some nonspecific proteins in normal human serum to complement, i.e., augment the effects of

The term complement refers to the ability of a system of some nonspecific proteins in normal human serum to complement, i.e., augment the effects of COMPLEMENT SYSTEM The term complement refers to the ability of a system of some nonspecific proteins in normal human serum to complement, i.e., augment the effects of other components of immune system,

More information

E and the heart: Possible role as antioxidant. Acta Vitaminol. Enzymol. 5: 11-22, ) Jolly, S. R., Kane, W. J., Bailie, M. B. et al.

E and the heart: Possible role as antioxidant. Acta Vitaminol. Enzymol. 5: 11-22, ) Jolly, S. R., Kane, W. J., Bailie, M. B. et al. 1) Ferrari, R., Visoli, O., Guarnieri, C. et al.: Vitamin E and the heart: Possible role as antioxidant. Acta Vitaminol. Enzymol. 5: 11-22, 1983. 2) Jolly, S. R., Kane, W. J., Bailie, M. B. et al.: Canine

More information

Spin-trapping detection of superoxides in polymorphonuclear leukocytes stimulated with serum-opsonized zymosan

Spin-trapping detection of superoxides in polymorphonuclear leukocytes stimulated with serum-opsonized zymosan Title Spin-trapping detection of superoxides in polymorpho Author(s)KUWABARA, Mikinori; TAKAHASHI, Tsuneo A.; NAGAHATA, CitationJapanese Journal of Veterinary Research, 48(1): 3-13 Issue Date 2000-05-31

More information

Human bronchoalveolar lavage cells and luminoldependent

Human bronchoalveolar lavage cells and luminoldependent J Clin Pathol 1981 ;34:167-171 Human bronchoalveolar lavage cells and luminoldependent chemiluminescence AJ WILLIAMS AND PJ COLE From the Host Defence Unit, Department of Medicine, Cardiothoracic Institute,

More information

Synovial Fluid Inhibits Killing of Staphylococcus aureus by Neutrophils

Synovial Fluid Inhibits Killing of Staphylococcus aureus by Neutrophils INFECTION AND IMMUNITY, June 1983, p. 14-11 Vol. 4, No. 3 19-9567/83/614-7$2./ Copyright C 1983, American Society for Microbiology Synovial Fluid Inhibits Killing of Staphylococcus aureus by Neutrophils

More information

CD B T NK NKT!! 1

CD B T NK NKT!! 1 CD B T NK NKT!! 1 2 !! 3 4 5 6 7 8 9 10 11 Biological effects of C5a 12 13 Opsonization and phagocytosis 14 15 http://www.med.sc.edu:85/book/wel come.htm 16 http://www.med.sc.edu:85/book/im munol-sta.htm

More information

Cellular & Molecular Immunology 2009

Cellular & Molecular Immunology 2009 Cellular & Molecular Immunology 2009 Complement Nicholas M. Ponzio, Ph.D. Department of Pathology & Laboratory Medicine March 4, 2009 Innate and adaptive immunity FAMOUS BELGIANS Jules Jean Baptiste Vincent

More information

Validation of the Efficacy of a Practical Method for Neutrophils Isolation from Peripheral Blood

Validation of the Efficacy of a Practical Method for Neutrophils Isolation from Peripheral Blood Validation of the Efficacy of a Practical Method for Neutrophils Isolation from Peripheral Blood JONATHAN DEGEL, MASIH SHOKRANI OBJECTIVE: The objectives of this study were to validate the Polymorphprep

More information

ANATOMY OF THE IMMUNE SYSTEM

ANATOMY OF THE IMMUNE SYSTEM Immunity Learning objectives Explain what triggers an immune response and where in the body the immune response occurs. Understand how the immune system handles exogenous and endogenous antigen differently.

More information

History. Chapter 13. Complement Components. Complement Pathways

History. Chapter 13. Complement Components. Complement Pathways History Chapter 13 Complement Jules Border in 1890 s discovered complement Paul Ehrlich coined the term complement The activity of blood serum that completes the action of antibody Now: Set of serum proteins

More information

Superoxide Dismutase Kit

Superoxide Dismutase Kit Superoxide Dismutase Kit Catalog Number: 7500-100-K Reagent kit for the analysis of Superoxide Dismutase in cell extracts. Sufficient reagents for 100 experimental tests, 50 negative controls, and 50 positive

More information

االستاذ المساعد الدكتور خالد ياسين الزاملي \مناعة \المرحلة الثانية \ التحليالت المرضية \ المعهد التقني كوت

االستاذ المساعد الدكتور خالد ياسين الزاملي \مناعة \المرحلة الثانية \ التحليالت المرضية \ المعهد التقني كوت Complement System The term complement refers to the ability of a system of some nonspecific proteins in normal human serum to complement, i.e., augment the effects of other components of immune system,

More information

Effect of temperature on liposome structures studied using EPR spectroscopy

Effect of temperature on liposome structures studied using EPR spectroscopy Spectroscopy 19 (2005) 37 42 37 IOS Press Effect of temperature on liposome structures studied using EPR spectroscopy W.W. Sułkowski a,,d.pentak a, W. Korus a and A. Sułkowska b a Department of Environmental

More information

Effect of Temperature on Bacterial Killing by Serum and by Polymorphonuclear Leukocytes

Effect of Temperature on Bacterial Killing by Serum and by Polymorphonuclear Leukocytes INFECTON AN ImmuNITy, June 1977, p. 947-954 Copyright 1977 American Societly for Microbiology Vol. 16, No. 3 Printed in U.S.A. Effect of Temperature on Bacterial Killing by Serum and by Polymorphonuclear

More information

Tetramethylpyrazine scavenges superoxide anion and decreases nitric oxide production in human polymorphonuclear leukocytes

Tetramethylpyrazine scavenges superoxide anion and decreases nitric oxide production in human polymorphonuclear leukocytes Life Sciences 72 (2003) 2465 2472 www.elsevier.com/locate/lifescie Tetramethylpyrazine scavenges superoxide anion and decreases nitric oxide production in human polymorphonuclear leukocytes Zhaohui Zhang

More information

Supporting Information. 58 Pages. 6 Figures 4 Tables

Supporting Information. 58 Pages. 6 Figures 4 Tables Light-Source-Dependent Effects of Main Water Constituents on Photodegradation of Phenicol Antibiotics: Mechanism and Kinetics Linke Ge, Jingwen Chen, * Xianliang Qiao, Jing Lin, Xiyun Cai Key Laboratory

More information

Complement: History. Discovered in 1894 by Bordet. It represents lytic activity of fresh serum

Complement: History. Discovered in 1894 by Bordet. It represents lytic activity of fresh serum Complement: History Discovered in 1894 by Bordet It represents lytic activity of fresh serum Its lytic activity destroyed when heated at 56C for 30 min Complement functions Host benefit: opsonization to

More information

Hydrogen Peroxide Production in Chronic Granulomatous Disease

Hydrogen Peroxide Production in Chronic Granulomatous Disease Hydrogen Peroxide Production in Chronic Granulomatous Disease A CYTOCHEMICAL STUDY OF REDUCED PYRIDINE NUCLEOTIDE OXIDASES RICHARD T. BRIGGS, MANFRED L. KARNOVSKY, and MoRms J. KARNOVSKY From the Departments

More information

Defense mechanism against pathogens

Defense mechanism against pathogens Defense mechanism against pathogens Immune System What is immune system? Cells and organs within an animal s body that contribute to immune defenses against pathogens ( ) Bacteria -Major entry points ;open

More information

HT Glutathione Assay Kit

HT Glutathione Assay Kit Instructions For Research Use Only. Not For Use In Diagnostic Procedures HT Glutathione Assay Kit Colorimetric assay for total, reduced and oxidized glutathione. Sufficient reagents for tests. Table of

More information

Citation Acta medica Nagasakiensia. 1997, 42

Citation Acta medica Nagasakiensia. 1997, 42 NAOSITE: Nagasaki University's Ac Title Review Article Children with Chroni Author(s) Tsuji, Yoshiro; Kondoh, Tatsuro; Qu Citation Acta medica Nagasakiensia. 1997, 42 Issue Date 1997-06-20 URL http://hdl.handle.net/10069/16071

More information

Development of Bactericidal Capacity and

Development of Bactericidal Capacity and INFECTION AND IMMUNITY, Feb. 1972, p. 232-237 Copyright 1972 American Society for Microbiology Vol. 5, No. 2 Printed in U.S.A. Development of Bactericidal Capacity and Phagocytosis-Associated Metabolism

More information

The measurement of superoxide anion production

The measurement of superoxide anion production European Journal of Clinical Investigation (1983) 13, 363-368 The measurement of superoxide anion production by granulocytes in whole blood. A clinical test for the evaluation of phagocyte function and

More information

PHAGOCYTOSIS BY MACROPHAGES

PHAGOCYTOSIS BY MACROPHAGES J. Cell Sci. 51, 189- aoi (1981) Printed in Great Britain Company of Biologists Limited 1981 PHAGOCYTOSIS BY MACROPHAGES II. THE DISSOCIATION OF THE ATTACHMENT AND INGESTION STEPS TADANAO ITO, MASAMICHI

More information

Oxidation of Escherichia coli Sulfhydryl Components by the Peroxidase-Hydrogen Peroxide-Iodide Antimicrobial System

Oxidation of Escherichia coli Sulfhydryl Components by the Peroxidase-Hydrogen Peroxide-Iodide Antimicrobial System ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, June 1978, p. 06-0066-4804/78/0013-06$02.00/0 Copyright 1978 American Society for Microbiology Vol. 13, No. 6 Printed in U.S.A. Oxidation of Escherichia coli Sulfhydryl

More information

TEST REPORT & SPECIFIC INFORMATION

TEST REPORT & SPECIFIC INFORMATION Page 1 (5) Dartsch Scientific GmbHAuf der Voßhardt 25 D-49419 Wagenfeld Firma LuKo Pharm GmbH Mayrwiesstrasse 25-27 A-5300 Hallwang Auf der Voßhardt 25 D-49419 Wagenfeld, Germany Fon: +49 5444 980 1322

More information

Overview of the immune system

Overview of the immune system Overview of the immune system Immune system Innate (nonspecific) 1 st line of defense Adaptive (specific) 2 nd line of defense Cellular components Humoral components Cellular components Humoral components

More information

ACTG Laboratory Technologist Committee Revised Version 2.0 ACTG Lab Man Coulter HIV-1 p24 ELISA May 21, 2004

ACTG Laboratory Technologist Committee Revised Version 2.0 ACTG Lab Man Coulter HIV-1 p24 ELISA May 21, 2004 Coulter HIV p24 1. PRINCIPLE The Human Immunodeficiency Virus Type 1 (HIV-1) is recognized as the etiologic agent of acquired immunodeficiency syndrome (AIDS). The virus is transmitted by sexual contact,

More information

Procaspase-3. Cleaved caspase-3. actin. Cytochrome C (10 M) Z-VAD-fmk. Procaspase-3. Cleaved caspase-3. actin. Z-VAD-fmk

Procaspase-3. Cleaved caspase-3. actin. Cytochrome C (10 M) Z-VAD-fmk. Procaspase-3. Cleaved caspase-3. actin. Z-VAD-fmk A HeLa actin - + + - - + Cytochrome C (1 M) Z-VAD-fmk PMN - + + - - + actin Cytochrome C (1 M) Z-VAD-fmk Figure S1. (A) Pan-caspase inhibitor z-vad-fmk inhibits cytochrome c- mediated procaspase-3 cleavage.

More information

Extracellular enzymes associated with lignin degradation

Extracellular enzymes associated with lignin degradation Reprinted from Biochemistry, 1990, 29. 10475 1990 by the American Chemical Society and reprinted by permission of the copyright owner. Lignin Peroxidase Oxidation of Mn 2+ in the Presence of Veratryl Alcohol,

More information

Bactericidal Mechanisms of Human Breast Milk Leukocytes

Bactericidal Mechanisms of Human Breast Milk Leukocytes INFECTION AND IMMUNITY, May 198, p. 314-318 19-9567/8/5-314/5$2./ Vol. 28, No. 2 Bactericidal Mechanisms of Human Breast Milk Leukocytes DIANNE F. JOHNSON, GENE L. FRANCE, DANIEL J. MARMER, AND RUSSELL

More information

Complement. History. Chapter 7. Complement Components. Complement Pathways. Pathways of complement activation

Complement. History. Chapter 7. Complement Components. Complement Pathways. Pathways of complement activation History Chapter 7 Complement Jules Border in 1890 s discovered complement Paul Ehrlich coined the term complement The activity of blood serum that completes the action of antibody Now: Set of serum proteins

More information

Laboratory 10 Factors Controlling Phagocytosis in Tetrahymena,

Laboratory 10 Factors Controlling Phagocytosis in Tetrahymena, BIO354: Cell Biology Laboratory 1 Laboratory 1 Factors Controlling Phagocytosis in Tetrahymena, I. Introduction A characteristic feature of all eukaryotic cells is the ability to pinch off portions of

More information

RETINOID-PHOSPHOLIPID INTERACTIONS AS STUDIED BY MAGNETIC RESONANCE. Stephen R. Wassail* and William Stillwellt

RETINOID-PHOSPHOLIPID INTERACTIONS AS STUDIED BY MAGNETIC RESONANCE. Stephen R. Wassail* and William Stillwellt Vol.''% No. 3 85 RETINOID-PHOSPHOLIPID INTERACTIONS AS STUDIED BY MAGNETIC RESONANCE Stephen R. Wassail* and William Stillwellt Departments of Physics* and Biology+ Indiana University-Purdue University

More information

Iron Chelates and Unwanted Biological Oxidations

Iron Chelates and Unwanted Biological Oxidations The Virtual Free Radical School Iron Chelates and Unwanted Biological Oxidations Kevin D. Welch and Steven D. Aust Department of Chemistry and Biochemistry Biotechnology Center Utah State University Logan,

More information

History. Chapter 13. Complement Components. Complement Pathways

History. Chapter 13. Complement Components. Complement Pathways History Chapter 13 Complement Jules Border in 1890 s discovered complement Paul Ehrlich coined the term complement The activity of blood serum that completes the action of antibody Now: Set of serum proteins

More information

OxiSelect Hydrogen Peroxide Assay Kit (Colorimetric)

OxiSelect Hydrogen Peroxide Assay Kit (Colorimetric) Product Manual OxiSelect Hydrogen Peroxide Assay Kit (Colorimetric) Catalog Number STA-343 5 assays FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Oxidative stress is a physiological

More information

Enzyme Action: Testing Catalase Activity

Enzyme Action: Testing Catalase Activity Enzyme Action: Testing Catalase Activity LabQuest 6A Many organisms can decompose hydrogen peroxide (H 2 O 2 ) enzymatically. Enzymes are globular proteins, responsible for most of the chemical activities

More information

RECEPTORS FOR C3b AND C3bi PROMOTE PHAGOCYTOSIS BUT NOT THE RELEASE OF TOXIC OXYGEN FROM

RECEPTORS FOR C3b AND C3bi PROMOTE PHAGOCYTOSIS BUT NOT THE RELEASE OF TOXIC OXYGEN FROM RECEPTORS FOR C3b AND C3bi PROMOTE PHAGOCYTOSIS BUT NOT THE RELEASE OF TOXIC OXYGEN FROM HUMAN PHAGOCYTES* BY SAMUEL D. WRIGHT AND SAMUEL C. SILVERSTE1N From the Laboratory for Cellular Physiology and

More information

Phagocytosis of FITC labelled opsonized and non-opsonized E. coli bacteria by monocytes and granulocytes in a whole blood assay

Phagocytosis of FITC labelled opsonized and non-opsonized E. coli bacteria by monocytes and granulocytes in a whole blood assay Phagocytosis of FITC labelled opsonized and non-opsonized E. coli bacteria by monocytes and granulocytes in a whole blood assay APPLICATION NOTE Author: Andreas Spittler, MD, Associate Professor for Pathophysiology

More information

Superoxide Dismutase Assay Kit

Superoxide Dismutase Assay Kit Superoxide Dismutase Assay Kit Catalog Number KA3782 100 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 General Information...

More information

Functional Analysis of Polymorphonuclear Leukocytes in Siblings of Glucose-6- Phosphate Dehydrogenase Deficiency*)

Functional Analysis of Polymorphonuclear Leukocytes in Siblings of Glucose-6- Phosphate Dehydrogenase Deficiency*) Hiroshima Journal of Medical Sciences Vol. 32, N.:>. 2, June, 1983 HIJM 32-27 173 Functional Analysis of Polymorphonuclear Leukocytes in Siblings of Glucose-6- Phosphate Dehydrogenase Deficiency*) Ryoji

More information

Supplementary Information: Liquid-liquid phase coexistence in lipid membranes observed by natural abundance 1 H 13 C solid-state NMR

Supplementary Information: Liquid-liquid phase coexistence in lipid membranes observed by natural abundance 1 H 13 C solid-state NMR Electronic Supplementary Material (ESI) for Physical Chemistry Chemical Physics. This journal is the wner Societies 28 Supplementary Information: Liquid-liquid phase coexistence in lipid membranes observed

More information

Simultaneous Blood Brain Barrier Crossing and Protection for Stroke Treatment Based on Edaravone-Loaded Ceria Nanoparticles

Simultaneous Blood Brain Barrier Crossing and Protection for Stroke Treatment Based on Edaravone-Loaded Ceria Nanoparticles Supporting Information Simultaneous Blood Brain Barrier Crossing and Protection for Stroke Treatment Based on Edaravone-Loaded Ceria Nanoparticles Qunqun Bao, Ping Hu, * Yingying Xu, Tiansheng Cheng, Chenyang

More information

Enzyme Action: Testing Catalase Activity

Enzyme Action: Testing Catalase Activity Enzyme Action: Testing Catalase Activity Pennsylvania Science Standards: S11.A.1.1.4 S11.A.1.3.1 S11.A.2.2.2.1 S11.A.2.2.2.2 Keystone Eligible Content Bio.B.4.1.1, Bio.B.4.1.2, and Bio.B.4.2.5 Introduction

More information

IMMUNITY AND ANTIBODIES

IMMUNITY AND ANTIBODIES IMMUNITY AND ANTIBODIES Stem cells in bone marrow differentiate into various blood cells Phagocytes attack alien cells A non-specific reaction Mast cells release histamine Histamine dilates capillaries,

More information

THE COMPLEMENT SYSTEM OBJECTIVES:

THE COMPLEMENT SYSTEM OBJECTIVES: Dr Mohammed Al- ani THE COMPLEMENT SYSTEM OBJECTIVES: When you finish this section, you should be able to: 1. Describe the effects of complement activation. 2. Outline the Classical, Mannan-Binding (MB)

More information

Question 1. Kupffer cells, microglial cells and osteoclasts are all examples of what type of immune system cell?

Question 1. Kupffer cells, microglial cells and osteoclasts are all examples of what type of immune system cell? Abbas Chapter 2: Sarah Spriet February 8, 2015 Question 1. Kupffer cells, microglial cells and osteoclasts are all examples of what type of immune system cell? a. Dendritic cells b. Macrophages c. Monocytes

More information

Heparin Sodium ヘパリンナトリウム

Heparin Sodium ヘパリンナトリウム Heparin Sodium ヘパリンナトリウム Add the following next to Description: Identification Dissolve 1 mg each of Heparin Sodium and Heparin Sodium Reference Standard for physicochemical test in 1 ml of water, and

More information

Osmotic resistance of heat-damaged erythrocytes

Osmotic resistance of heat-damaged erythrocytes J. clin. Path. (1967), 20, 239 S. BAAR From the M. R. C. Industrial Injuries and Burns Research Unit, Birmingham Accident Hospital SYNOPSIS Whole blood was heated for twenty minutes at 40 C., 45 C., 500C.,

More information

Name: Date: AP Biology LAB : FACTORS INFLUENCING ENZYME ACTIVITY

Name: Date: AP Biology LAB : FACTORS INFLUENCING ENZYME ACTIVITY LAB : FACTORS INFLUENCING ENZYME ACTIVITY Background Enzymes are biological catalysts capable of speeding up chemical reactions by lowering activation energy. One benefit of enzyme catalysts is that the

More information

BIOCHEMISTRY OF BLOOD

BIOCHEMISTRY OF BLOOD BCH 471 BIOCHEMISTRY OF BLOOD Amal Alamri Experiment 1 Separation of Plasma and Serum from Whole Blood Whole Blood It is living tissue that circulates through the heart, arteries, veins, and capillaries

More information

See external label 2 C-8 C = C-REACTIVE PROTEIN (CRP) LATEX SLIDE TEST

See external label 2 C-8 C = C-REACTIVE PROTEIN (CRP) LATEX SLIDE TEST CORTEZ DIAGNOSTICS, INC. 21250 Califa Street, Suite 102 and 116, Woodland Hills, CA 91367 Tel: (818) 591-3030 Fax: (818) 591-8383 onestep@rapidtest.com technicalsupport@rapidtest.com www.rapidtest.com

More information

Effects of Catechins on Superoxide and Hydroxyl Radical

Effects of Catechins on Superoxide and Hydroxyl Radical February 1999 Chem. Pharm. Bull. 47(2) 279 283 (1999) 279 Effects of Catechins on Superoxide and Hydroxyl Radical Midori KASHIMA Department of Endodontics, Nihon University School of Dentistry at Matsudo

More information

Plasma Hemoglobin Determination: Two Manual Methods Compared. An Honors Thesis (HONRS 499) Anne M. Emerick. Sharlene Strahl. Beech Grove, Indiana

Plasma Hemoglobin Determination: Two Manual Methods Compared. An Honors Thesis (HONRS 499) Anne M. Emerick. Sharlene Strahl. Beech Grove, Indiana Plasma Hemoglobin Determination: Two Manual Methods Compared An Honors Thesis (HONRS 499) by Anne M. Emerick Sharlene Strahl St. Francis Hospital and Health Centers Beech Grove, Indiana Dr. Nancy Behforouz

More information

Superoxide Dismutase Kit

Superoxide Dismutase Kit Superoxide Dismutase Kit Catalog Number: 7500-100-K Reagent kit for the analysis of Superoxide Dismutase in cell extracts. Sufficient reagents for 100 experimental tests, 50 negative controls, and 50 positive

More information

BIOL 305L Spring 2019 Laboratory Six

BIOL 305L Spring 2019 Laboratory Six Please print Full name clearly: BIOL 305L Spring 2019 Laboratory Six Osmosis in potato and carrot samples Introduction Osmosis is the movement of water molecules through a selectively permeable membrane

More information

Immunologic Cross-Reaction Between Luteinizing Hormone and Human Chorionic Gonadotropin

Immunologic Cross-Reaction Between Luteinizing Hormone and Human Chorionic Gonadotropin Immunologic Cross-Reaction Between Luteinizing Hormone and Human Chorionic Gonadotropin MELVIN L. TAYMOR, M.D., DONALD A. GOSS, M.D., and ALBERT BUYTENDORP, M.D. RECENTLY a number of reports 2 4 have indicated

More information

Innate Immunity. Bởi: OpenStaxCollege

Innate Immunity. Bởi: OpenStaxCollege Innate Immunity Bởi: OpenStaxCollege The vertebrate, including human, immune system is a complex multilayered system for defending against external and internal threats to the integrity of the body. The

More information

ISOLATION OF EXFOLIATED SOMATIC CELLS FROM BUFFALO MILK

ISOLATION OF EXFOLIATED SOMATIC CELLS FROM BUFFALO MILK Original Article Buffalo Bulletin (March 2013) Vol.32 No.1 ISOLATION OF EXFOLIATED SOMATIC CELLS FROM BUFFALO MILK A.K. Dang*, Joydip Mukherjee, Manu Jamwal, Surender Singh, A.K. Mohanty, Shiv Prasad,

More information

Hahn Lab Dye Kit. Contents of dye kit:

Hahn Lab Dye Kit. Contents of dye kit: Hahn Lab Dye Kit Contents of dye kit: dye name donor/acceptor code references (see below) Mero53 si-so-ia 4 Mero58 TD-BA-sIA - Mero59 TD-SO-sIA - Mero60 I-Pht-sIA 1 Mero61 I-BA-sIA 1 Mero62 I-TBA-sIA 1

More information

Composition of Blood

Composition of Blood Blood is a connective tissue, specialized to transport the respiratory gasses as well as hormones, nutrients, and wastes, and the distribution of heat. The various cells of the blood perform specific functions.

More information

CELL MEDIATED IMMUNE RESPONSE

CELL MEDIATED IMMUNE RESPONSE CELL MEDIATED IMMUNE RESPONSE Chapter IV - CELL MEDIATED IMMUNE RESPONSE Sujatha, M. 2013. Evaluation of Immunological changes in Fish, Catla catla administered with bacterial pathogen, Aeromonas hydrophila,

More information

HT Glutathione Assay Kit

HT Glutathione Assay Kit IFU0 Rev Status: RELEASED printed //0 ::0 AM by Trevigen Document Control Instructions For Research Use Only. Not For Use In Diagnostic Procedures HT Glutathione Assay Kit Colorimetric assay for total,

More information

CONTENTS. STUDY DESIGN METHODS ELISA protocol for quantitation of mite (Dermatophagoides spp.) Der p 1 or Der f 1

CONTENTS. STUDY DESIGN METHODS ELISA protocol for quantitation of mite (Dermatophagoides spp.) Der p 1 or Der f 1 CONTENTS STUDY DESIGN METHODS ELISA protocol for quantitation of mite (Dermatophagoides spp.) Der p 1 or Der f 1 ELISA protocol for mite (Dermatophagoides spp.) Group 2 ALLERGENS RESULTS (SUMMARY) TABLE

More information

Role of the Phagocyte in Host-Parasite Interactions

Role of the Phagocyte in Host-Parasite Interactions INFECTION AND IMMUNITY, OCt. 1970, p. 414-418 Copyright 1970 American Society for Microbiology Vol. 2, No. 4 Printed in U.S.A. Role of the Phagocyte in Host-Parasite Interactions XXIV. Aldehyde Generation

More information

Catalog Number: A114 Sizes Available: 250 µg/vial Concentration: 1.0 mg/ml (see Certificate of Analysis for actual concentration)

Catalog Number: A114 Sizes Available: 250 µg/vial Concentration: 1.0 mg/ml (see Certificate of Analysis for actual concentration) Name: C3b Catalog Number: A114 Sizes Available: 250 µg/vial Concentration: 1.0 mg/ml (see Certificate of Analysis for actual concentration) Form: Liquid Purity: >90% by SDS-PAGE Buffer: 10 mm sodium phosphate,

More information

Title. YOON, Seokjoo; MARUYAMA, Yutaka; KA FUJITA, Shoichi. Author(s) Issue Date /jjvr

Title. YOON, Seokjoo; MARUYAMA, Yutaka; KA FUJITA, Shoichi. Author(s) Issue Date /jjvr Title Application of FT-IR and ESR spectr study of CCl_4-induced peroxidation Author(s) YOON, Seokjoo; MARUYAMA, Yutaka; KA FUJITA, Shoichi Citation Japanese Journal of Veterinary Rese Issue Date 2000-02-29

More information

Hydrogen Peroxide Release by Rat Alveolar Macrophages: Comparison with Blood Neutrophils

Hydrogen Peroxide Release by Rat Alveolar Macrophages: Comparison with Blood Neutrophils NFECTON AND MMUNTY, Feb. 1978, p. 621-629 0019-9567/78/0019-0621$02.00/0 Copyright X 1978 American Society for Microbiology Vol. 19, No. 2 Printed in U.S.A. Hydrogen Peroxide Release by Rat Alveolar Macrophages:

More information

Efflux of Red Cell Water into Buffered Hypertonic Solutions

Efflux of Red Cell Water into Buffered Hypertonic Solutions Efflux of Red Cell Water into Buffered Hypertonic Solutions EDWIN G. OLMSTEAD From the School of Medicine, University of North Dakota, Grand Forks ABSTRACT Buffered NaCI solutions hypertonic to rabbit

More information

INVESTIGATION OF CHLORINATED METHANES TREATABILITY USING ACTIVATED SODIUM PERSULFATE

INVESTIGATION OF CHLORINATED METHANES TREATABILITY USING ACTIVATED SODIUM PERSULFATE Preprint: Proceedings of the First International Conference on Environmental Science and Technology (2005) INVESTIGATION OF CHLORINATED METHANES TREATABILITY USING ACTIVATED SODIUM PERSULFATE Duane K.

More information

IN VITRO CELLULAR RESPONSES TO AUTOLOGOUS TUMOR EXTRACT DETECTED BY INHIBITION OF MACROPHAGE MIGRATION*1

IN VITRO CELLULAR RESPONSES TO AUTOLOGOUS TUMOR EXTRACT DETECTED BY INHIBITION OF MACROPHAGE MIGRATION*1 [Gann, 66, 167-174; April, 1975] IN VITRO CELLULAR RESPONSES TO AUTOLOGOUS TUMOR EXTRACT DETECTED BY INHIBITION OF MACROPHAGE MIGRATION*1 Tsuyoshi AKIYOSHI, Akira HATA, and Hideo TSUJI Department of Surgery,

More information

Stimulation of Human and Canine Neutrophil Metabolism by

Stimulation of Human and Canine Neutrophil Metabolism by ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Feb. 1981, p. 284-289 0066-4804/81/020284-06$02.00/0 Vol. 19, No. 2 Stimulation of Human and Canine Neutrophil Metabolism by Amphotericin B SEE-RUERN SUPAPIDHAYAKUL,'2

More information

CYTOCHEMICAL DEMONSTRATION OF HYDROGEN PEROXIDE IN

CYTOCHEMICAL DEMONSTRATION OF HYDROGEN PEROXIDE IN Published Online: 1 January, 1975 Supp Info: http://doi.org/10.1083/jcb.64.1.254 Downloaded from jcb.rupress.org on June 30, 2018 CYTOCHEMICAL DEMONSTRATION OF HYDROGEN PEROXIDE IN POLYMORPHONUCLEAR LEUKOCYTE

More information

Issued by: LABORATORY MANAGER Original Date: March 20, 2000 Approved by: Laboratory Director Revision Date: July 26, 2000 CRYPTOCOCCAL ANTIGEN

Issued by: LABORATORY MANAGER Original Date: March 20, 2000 Approved by: Laboratory Director Revision Date: July 26, 2000 CRYPTOCOCCAL ANTIGEN Policy # MI\TECH\11\v01 Page 1 of 6 Issued by: LABORATORY MANAGER Original Date: March 20, 2000 Approved by: Laboratory Director Revision Date: July 26, 2000 CRYPTOCOCCAL ANTIGEN Latex particles coated

More information

Superoxide Dismutase Microplate Assay Kit User Manual

Superoxide Dismutase Microplate Assay Kit User Manual Superoxide Dismutase Microplate Assay Kit User Manual Catalog # CAK1010 Detection and Quantification of Superoxide Dismutase (SOD) Activity in Urine, Serum, Plasma, Tissue extracts, Cell lysate, Cell culture

More information

Innate Immunity Part I October 3, Dan Stetson

Innate Immunity Part I October 3, Dan Stetson Innate Immunity Part I October 3, 2016 Dan Stetson stetson@uw.edu 441 Lecture #3 Slide 1 of 28 Three lectures on innate Immunity Part 1 (Today): Introduction and concepts Overview of main components and

More information

The Immune System. Specific Immunity

The Immune System. Specific Immunity The Immune System Specific Immunity What You Should Know Immune surveillance A range of white blood cells constantly circulate monitoring the tissues. If tissues become damaged or invaded, cells release

More information

Investigating Ribonuclease A Modification Induced by Quinones Using UV/Vis Spectroscopy

Investigating Ribonuclease A Modification Induced by Quinones Using UV/Vis Spectroscopy Investigating Ribonuclease A Modification Induced by Quinones Using UV/Vis Spectroscopy Caitlin Redman, Albert Vaughn, Steve Ledford and Jisook Kim, University of Tennessee at Chattanooga, Chattanooga,

More information

ENZYMES QUESTIONSHEET 1

ENZYMES QUESTIONSHEET 1 QUESTIONSHEET 1 The apparatus illustrated below can be used to investigate the activity of the enzyme catalase, which is found in liver. The liver tissue has been ground up and mixed with a buffer solution.

More information

Hematopoiesis. Hematopoiesis. Hematopoiesis

Hematopoiesis. Hematopoiesis. Hematopoiesis Chapter. Cells and Organs of the Immune System Hematopoiesis Hematopoiesis- formation and development of WBC and RBC bone marrow. Hematopoietic stem cell- give rise to any blood cells (constant number,

More information

Superoxide Dismutase Activity in Leukocytes

Superoxide Dismutase Activity in Leukocytes Superoxide Dismutase Activity in Leukocytes LAWRENCE R. DECHATELET, CHARLEs E. MCCALL, LnDmA C. MCPHAIL, and RICHARD B. JOHNSTON, JR. From the Departments of Biochemistry and Medicine, The Bowman Gray

More information

Animal model for testing human Ascaris allergens

Animal model for testing human Ascaris allergens J. Biosci., Vol. 3 Number 1, March 1981, pp. 77-82. Printed in India. Animal model for testing human Ascaris allergens KRISHNA MUKERJI*, R. P. SAXENA, S. N. GHATAK and K. C. SAXENA Division of Biochemistry,

More information

WJEC. Respiration. Questions

WJEC. Respiration. Questions WJEC Respiration Questions 6. Answer one of the following questions. Any diagrams included in your answer must be fully annotated. 13 Examiner only Arholwr yn unig Either, (a)

More information

Complete Deficiency of Leukocyte Glucose-6-Phosphate Dehydrogenase with Defective Bactericidal Activity

Complete Deficiency of Leukocyte Glucose-6-Phosphate Dehydrogenase with Defective Bactericidal Activity Complete Deficiency of Leukocyte Glucose-6-Phosphate Dehydrogenase with Defective Bactericidal Activity M. ROBERT COOPER, LAWRENCE R. DECHATELET, CHARLEs E. MCCALL, MARIANO F. LAVIA, CHARLES L. SPuRR,

More information

Apoptosis of lymphocytes in SLE: the level, correlation with dosage prednisolone and lymphocyte phenotypes

Apoptosis of lymphocytes in SLE: the level, correlation with dosage prednisolone and lymphocyte phenotypes Apoptosis of lymphocytes in SLE: the level, correlation with dosage prednisolone and lymphocyte phenotypes ABDELMAROUF MOHIELDEIN 1, NATALIA BELUSHKINA 2, ULIANA PETROVA 3. 1,2 Department of Biochemistry,

More information

A 207c solution by weight of yeast was used throughout the study. This. Enzymatic Action in the Presence of Some Common Antiseptics

A 207c solution by weight of yeast was used throughout the study. This. Enzymatic Action in the Presence of Some Common Antiseptics Enzymatic Action in the Presence of Some Common Antiseptics O. E. Rumple and R. J. Hartman, Indiana University Many diversified studies of the influence of foreign materials on enzymatic action have been

More information

Resisting infection. Cellular Defenses: Leukocytes. Chapter 16: Innate host defenses Phagocytosis Lymph Inflammation Complement

Resisting infection. Cellular Defenses: Leukocytes. Chapter 16: Innate host defenses Phagocytosis Lymph Inflammation Complement Resisting infection Chapter 16: Innate host defenses Lymph Inflammation Complement Bio 139 Dr. Amy Rogers Innate defenses (ch. 16) Physical & chemical barriers; cellular defenses; inflammation, fever;

More information

An improved technique for the assay of red blood cell superoxide dismutase (SOD) activity

An improved technique for the assay of red blood cell superoxide dismutase (SOD) activity ELSEVIER Clinica Chimica Acta 247 (1996) 1-6 An improved technique for the assay of red blood cell superoxide dismutase (SOD) activity L.E. Ilouno, E.N. Shu*, G.E. Igbokwe Department of Applied Biochemistry,

More information

Investigating an immunocompromised child

Investigating an immunocompromised child Current Practice Investigating an immunocompromised child Anura Weerasinghe l Sri Lanka Journal of Child Health, 2000; 29:116-9 (Key words: immunocompromised child, investigation) Introduction The term

More information