Oxidative Metabolic Responses Research Articles

Respiratory performance and behavioral responses of the freshwater fish, Cyprinus carpio (Linnaeus) under sublethal exposure to chlorpyrifos

A short-term definitive test by static renewal bioassay method was conducted to determine the acute toxicity (LC50) of an organophosphate insecticide, chlorpyrifos on the fish, Cyprinus carpio. Carp fingerlings were exposed to different concentrations (0.120 to 0.200 mg/L) of chlorpyrifos for 96 h. The acute toxicity of chlorpyrifos was found to be 0.160 m g/L. One-seventh (1/7th, 0.0224 mg/L) and one-fourteenth (1/14th, 0.0112 mg/L) of the acute toxicity value were selected as sublethal concentrations for subchronic studies. The fish were exposed to both the sublethal concentrations for 1, 7, and 14 days and allowed to recover in toxicant-free medium for 7 days. Behavioral responses and respiratory rate were studied in experimental periods. Fish in toxic media exhibited irregular, erratic, and darting swimming movements, hyperexcitability, and loss of equilibrium and sinking to the bottom. Caudal bending was the chief morphological alterations during the exposure tenures. The behavioral and morphologic changes might be due to the inhibition of acetylcholinesterase (AChE) activity. The carp were found under stress but mortality was insignificant at both the sublethal concentrations. Considerable variation in respiratory rates (-71.85 to 119.52%; -75.65 to 17.58%) were observed in the 1/7th and 1/14th of lethal concentration of chlorpyrifos respectively. The alteration in respiratory rate is due to respiratory distress, this may be a consequence of impaired oxidative metabolism and elevated physiological response during chlorpyrifos exposure. The impairments in fish respiratory physiology and behavioral responses even under recovery periods may be due to slow release of sequestered chlorpyrifos from storage tissues.

Structure of the Noncatalytic Domains and Global Fold of the Protein Disulfide Isomerase ERp72

Protein disulfide isomerases are a family of proteins that catalyze the oxidation and isomerization of disulfide bonds in newly synthesized proteins in the endoplasmic reticulum. The family includes general enzymes such as PDI that recognize unfolded proteins, and others that are selective for specific classes of proteins. Here, we report the X-ray crystal structure of central non-catalytic domains of a specific isomerase, ERp72 (also called CaBP2 and protein disulfide-isomerase A4) from Rattus norvegicus. The structure reveals strong similarity to ERp57, a PDI-family member that interacts with the lectin-like chaperones calnexin and calreticulin but, unexpectedly, ERp72 does not interact with calnexin as shown by isothermal titration calorimetry and nuclear magnetic resonance (NMR) spectroscopy. Small-angle X-ray scattering (SAXS) of ERp72 was used to develop models of the full-length protein using both rigid body refinement and ab initio simulated annealing of dummy atoms. The two methods show excellent agreement and define the relative positions of the five thioredoxin-like domains of ERp72 and potential substrate or chaperone binding sites.

UCP3 Expression in Liver Modulates Gene Expression and Oxidative Metabolism in Response to Fatty Acids, and Sensitizes Mitochondria to Permeability Transition

Background/Aims: Uncoupling protein-3 (UCP3) is expressed in liver only under conditions of high fatty acid catabolism. However, the specific role of UCP3 in liver mitochondria and overall hepatic function is still poorly known. Methods: A model of “in vivo” induction of UCP3 expression in mouse liver mitochondria via a tail-vein injection of a recombinant adenoviral vector was developed. The effects on liver mitochondrial bioenergetics and permeability transition, liver gene expression, and systemic metabolism were then determined. Results: UCP3 expression in liver did not cause basal, non-specific, uncoupling but led to a stimulation of palmitate-induced state 4 respiration. UCP3 expression in liver also caused an increase in the expression of certain genes involved in lipid catabolism and metabolic response to starvation (e.g. medium chain acyl-CoA-dehydrogenase or peroxisome proliferator-activated receptor-γ co-activator-1α). UCP3 also conferred to liver mitochondria an enhanced sensitivity to classical inducers of permeability transition, such as calcium and carboxyatractylate. Conclusion: UCP3 expression in liver exerts direct actions on mitochondrial activity, favoring fatty acid-induced uncoupling and sensitizing mitochondria to permeability transition, as well as causing retrograde signaling to nuclear gene expression consistent with favoring lipid catabolism and oxidative metabolism.

An update on iron physiology

Iron is an essential micronutrient, as it is required for adequate erythropoietic function, oxidative metabolism and cellular immune responses. Although the absorption of dietary iron (1-2 mg/d) is regulated tightly, it is just balanced with losses. Therefore, internal turnover of iron is essential to meet the requirements for erythropoiesis (20-30 mg/d). Increased iron requirements, limited external supply, and increased blood loss may lead to iron deficiency (ID) and iron-deficiency anemia. Hepcidin, which is made primarily in hepatocytes in response to liver iron levels, inflammation, hypoxia and anemia, is the main iron regulatory hormone. Once secreted into the circulation, hepcidin binds ferroportin on enterocytes and macrophages, which triggers its internalization and lysosomal degradation. Thus, in chronic inflammation, the excess of hepcidin decreases iron absorption and prevents iron recycling, which results in hypoferremia and iron-restricted erythropoiesis, despite normal iron stores (functional ID), and anemia of chronic disease (ACD), which can evolve to ACD plus true ID (ACD + ID). In contrast, low hepcidin expression may lead to iron overload, and vice versa. Laboratory tests provide evidence of iron depletion in the body, or reflect iron-deficient red cell production. The appropriate combination of these laboratory tests help to establish a correct diagnosis of ID status and anemia.

Skeletal Muscle and Liver Oxidative Metabolism in Response to a Voluntary Isocaloric Intake of a High Fat Diet in Male and Female Rats

High fat diets (HFD) usually lead to hyperphagia and body weight gain. However, macronutrient proportions in the diet can modulate energy intake and body fat deposition. The aim of the study was to investigate muscle and liver oxidative metabolism in response to an isocaloric intake of a HFD and to elucidate the possible gender-dependent response. Eight week-old male and female rats were fed either standard chow or HFD for 14 weeks. Energy intake, body weight and whole animal oxygen consumption were determined periodically. Mitochondrial oxygen consumption, hydrogen peroxide production, TBARS levels, Cytochrome-c-oxidase, Citrate synthase and antioxidant enzyme activities were measured in muscle and liver. UCP1 and UCP3 protein levels were analyzed in brown adipose tissue and muscle, respectively. Male rats showed higher energy efficiency, enhanced adiposity, greater hydrogen peroxide production and less effective antioxidant machinery compared to females. HFD feeding increased energy expenditure but did not modify either tissue oxidative metabolism or oxidative damage in either gender. HFD animals over-expressed uncoupling proteins in order to maintain energy balance (brown adipose tissue UCP1) and to avoid oxidative stress (skeletal muscle UCP3), thus counteracting the alterations induced by the modification of the proportion of macronutrients in the diet.

Nitrogen nutrition and antioxidant metabolism in ammonium‐tolerant and ‐sensitive plants

Ammonium nutrition is of interest as an alternative to that of using nitrate. However, the former has been reported as stressful to many plant species especially to some important crops, as most abiotic stresses may trigger oxidative imbalances in plants. In this work, we investigate the response of oxidative metabolism of two plant species, spinach (Spinacia oleracea L. cv. Gigante de invierno) and pea (Pisum sativum L. cv. Rondo), which have distinct tolerance to ammonium. Plants were grown in the presence of 1.5 and 3.0 mM N as ammonium and compared with equivalent nitrate nutrition. The antioxidant enzymes and metabolites as well as oxidative damage to proteins were determined. Protein and amino acid contents in both types of plants were also analysed. Ammonium nutrition in sensitive spinach or in the tolerant pea plants does not alter the redox status of ascorbate and glutathione or the phenolic contents, while no clear effect is seen in the antioxidant enzymes. The results showed that the stress originated from applying ammonium as the only N source is not an oxidative stress, independent of the ammonium tolerance of the plant species studied. Moreover, ammonium stress diminishes oxidative damage to proteins in the spinach plants. The data of the protein oxidation together with those from N metabolism highlight the relation between the stress induced by ammonium and an increased protein turnover.

Optical imaging of the spatiotemporal dynamics of cerebral blood flow and oxidative metabolism in the rat barrel cortex

Oxidative metabolism and cerebral blood flow (CBF) are two of the most important measures in neuroimaging. However, results from concurrent imaging of the two with high spatial and temporal resolution have never been published. We used flavoprotein autofluorescence (AF) and laser speckle imaging (LSI) in the anaesthetized rat to map oxidative metabolism and CBF in response to single vibrissa stimulation. Autofluorescence responses reflecting oxidative metabolism demonstrated a fast increase with a delay of 0.1 s. The sign-reversed speckle contrast reflecting CBF started to rise with a delay of 0.6 s and reached its maximum 1.4 s after the stimulation offset. The fractional signal changes were 2.0% in AF and 9.7% in LSI. Pixelwise modelling revealed that CBF maps spread over an area up to 2.5-times larger than metabolic maps. The results provide evidence that the increase in cerebral oxidative metabolism in response to sensory stimulation is considerably faster and more localized than the CBF response. This suggests that future developments in functional imaging concentrating on the metabolic response promise an increased spatial resolution.

Exposure to pressure stimulus enhances succinate dehydrogenase activity in L6 myoblasts.

Contraction of skeletal muscle generates pressure stimuli to intramuscular tissues. However, the effects of pressure stimuli, other than those created by electricity or nerve impulse, on physiological and biochemical responses in skeletal muscles are unknown. The purpose of this study is to examine the effects of a pure pressure stimulus on metabolic responses in a skeletal muscle cell line. Atmospheric pressure was applied to L6 myoblasts using an original apparatus. Succinate dehydrogenase (SDH) activity was evaluated by colorimetric assay using tetrazolium monosodium salt. The amounts of 2-deoxy-[(3)H]glucose uptake and lactate release were measured. SDH activity was 2.6- to 2.9-fold higher in pressurized L6 cells than in nonpressurized L6 cells (P < 0.01), and 2-deoxy-[(3)H]glucose uptake was 2.2-fold higher (P < 0.001). In addition, the amount of released lactate decreased from 6.8 to 3.7 mumol/dish when pressure was applied (P < 0.001). In contrast, the intracellular lactate contents of the pressurized cells were higher than those of nonpressurized cells (P < 0.01). However, the total amount of released lactate and intracellular lactate was lower in the pressurized cells than in nonpressurized cells. These findings demonstrate that a pure pressure stimulus enhances aerobic metabolism in L6 skeletal muscle cells and raise the possibility that elevated intramuscular pressure during muscle activity may be an important factor in stimulating oxidative metabolic responses in skeletal muscles.

Aerobic Exercise Dysfunction in Human Immunodeficiency Virus: A Potential Link to Physical Disability

Approximately 282,000 adults, adolescents, and children are currently living with human immunodeficiency virus (HIV) infection or acquired immunodeficiency syndrome (AIDS) in the United States as of the year 2002.1 With the advent of highly active antiretroviral therapy (HAART), the life span of these individuals has dramatically increased,2 and HIV infection is now considered a chronic illness with accompanying episodes of exacerbations and remissions of symptoms.3 Numerous conditions such as lipodystrophy syndrome4–6 and skeletal myopathy7–9 have been associated with HIV and its medical management, many of which may result in physical disability and diminished quality of life. Due to the chronic nature of this condition, physical therapists will continue to manage many of these conditions in increased numbers of people who are living with HIV. Although guidelines for physical therapy evaluation and management of this potentially disabling condition have not been established, it appears that aerobic exercise training may have a beneficial effect on the cardiorespiratory health of people who are living with HIV. Thus, an understanding of the factors that limit the oxidative metabolic response to physical activity is paramount in developing effective exercise training programs for people with this virus. The focus of this Update is to examine the biological factors that might limit the oxidative metabolic response to physical activity in people with HIV. HIV-related disability has been associated with fatigue and decreased physical functioning10–13 as well as other factors that may limit people's ability to carry out necessary life activities.12,14,15 Fatigue in adults with HIV has been identified using patient self-reports of physical and functional limitations while performing activities of daily living such as housework, climbing stairs, and walking15 and activities required for employment.10,11 Asymptomatic HIV …

PROAND ANTIOXIDANT EFFECT OF THERAPEUTIC MICROWAVE IRRADIATION (460 MHZ) ON BRAIN TISSUES IN EXPERIMENT

The article concerns biological effects of non-ionized electromagnetic irradiation which is considered in modern world as a serious ecological factor. Influence of decimetric microwave irradiation (460 MHz) on free radical processes in the rat brain tissues - visual cortex and hypothalamus - were investigated. Oxidative effects of both lipid peroxidation and specific activity of glutathione reductase were determined. It is shown that whole body irradiation for up to 4 weeks results in considerable changes of the above indices dependent on the irradiation intensity and age of the animals. Decimetric microwaves have oxidant effects at high intensity irradiation (SAR - 15 mW/kg), while these effects are antioxidant at low intensity (SAR - 5 mW/kg). Physiological implications of different oxidative metabolic responses of the hypothalamus and the cortex to microwaves irradiation are discussed.

Some magnesium status indicators and oxidative metabolism responses to low-dietary magnesium are affected by dietary copper in postmenopausal women

Objective A study with human volunteers was conducted to ascertain whether a low intake of copper (Cu) would exacerbate the response to a deficient intake of magnesium (Mg). Methods Nineteen postmenopausal women, age 47 to 78 y, completed a metabolic unit study as designed. For 162 d, nine women were fed a diet containing 1.0 mg of Cu/2000 kcal and 10 women were fed 3.0 mg of Cu/2000 kcal. Diets contained 99 or 399 mg of Mg/2000 kcal for 81 d in a randomized, double-blind, crossover design. Differences were considered significant when statistical analysis yielded P ≤ 0.05. Results Magnesium balance was highly positive when the dietary magnesium was high but non-positive when dietary magnesium was low. Copper balance was more positive when dietary copper was high than when it was low. Plasma ionized magnesium was decreased by magnesium deprivation. Several variables measured indicated that low dietary copper affected the response to magnesium deprivation or vice-versa. Red blood cell magnesium was lower when dietary copper was low than when it was high. When dietary magnesium was low, serum copper was lower in the women fed marginal copper than in those fed luxuriant copper. When dietary magnesium was high, low dietary copper did not affect serum copper. Magnesium deprivation decreased red blood cell superoxide dismutase when dietary copper was luxuriant; when dietary copper was low, magnesium deprivation did not have much of an effect. Apolipoprotein A1 was lowest when dietary magnesium and copper were low. The order in which the magnesium restriction occurred affected the response of a number of variables to this treatment including concentrations of serum magnesium and total and low-density lipoprotein cholesterol. Conclusions The findings indicated that, in short-term magnesium depletion experiments, the response to depletion can be influenced by other dietary factors including copper intake and a high magnesium intake before depletion, and that 100 mg of Mg/d is inadequate for postmenopausal women.

Response of oxidative metabolism in watermelon plants subjected to cold stress.

The objective of the present work was to determine the effect of thermal stress on oxidative metabolism in Citrullus lanatus [Thomb.] Mansf. cv. Dulce maravilla. Plants were grown for 30 d at two temperatures (10 and 35˚C), at which time we measured the leaf concentration of antioxidant compounds (ascorbate, dehydroascorbate, reduced glutathione, oxidized glutathione) and enzymatic activities [superoxide dismutase (SOD), catalase, guaiacol peroxidase, ascorbate peroxidase, dehydroascorbate reductase and glutathione reductase], as well as total hydrogen peroxide (H2O2) concentration and shoot dry weight. Our results indicate that chilling stress occurred in watermelon plants at 10˚C, while 35˚C is the optimal temperature for this plant. Low temperature stress caused: (i) decreased shoot weight; (ii) accumulation of H2O2; (iii) increased SOD activity; and (iv) decreased enzyme activities associated with detoxifying H2O2. The novelty of this study centres on the fact that so few cold-sensitive species have been examined to date - additional cold-sensitive species need to be studied to determine if there are shared characteristics in terms of how they respond to cold stress. Most studies have examined single antioxidant responses, whereas we conducted a comprehensive examination of many antioxidant responses.

Response of oxidative metabolism to the application of carbendazim plus boron in tobacco

In view of the essential role of oxidative process in the development of pathogen resistance in plants, the aim of the present study was to determine the individual effect of a fungicide, as well as to determine the combined effect of the fungicide and boron (B) on superoxide dismutase (SOD), guaiacol peroxidase (GPX), catalase (CAT), and ascorbate peroxidase (APX) activities and H 2 O 2 levels in tobacco plants (Nicotiana tabacum L. cv. Tennessee 86). The fungicide applied was carbendazim (carb) at a concentration of 2.6 mM . Boron was applied as H 3 BO 3 at: 1.6 mM (B1), 4 mM (B2), 8 mM (B3), 16 mM (B4), 32 mM (B5), and 64 mM (B6). The results indicated that foliar application of carbendazim by itself does not increase SOD, GPX, CAT or APX activities or H2 O2 foliar accumulation. The combined application of carbendazim and B increased SOD, GPX, CAT, and APX activities, especially in carb-B3 and carb-B4 . This effect may signify an additional tolerance mechanism to pathogenic infection, given the participation of these enzymes in the early phases of the plant–pathogen interactions.

Influence of caloric intake on aging and on the response to stressors

Reducing the food intake of rodents to well below that of ad libitum fed animals increases the life span. This action, which gerontologists often refer to as the antiaging action of dietary restriction (DR), is due to the slowing of the aging processes. DR also maintains most physiological processes in a youthful state and delays the occurrence and/or slows the progression of age‐associated disease processes. This antiaging action of DR results from the reduced intake of calories. Reduction of the body fat content does not play a causal role in the antiaging action of DR, nor does reduction in the metabolic rate. Alterations in the characteristics of carbohydrate metabolism and of oxidative metabolism in response to DR have been found that are of such a nature that they could, at least in part, underlie the antiaging action. Several theories have recently been proposed in regard to the mechanisms responsible for the antiaging action of DR, but none has been tested by rigorously designed studies. Of these theories, the one that seems most promising is based on the fact that DR protects rats and mice of all ages against the damaging actions of acute stressors. This protective action against stressors may play a major role in the antiaging action of DR.

Subthalamic ablation reverses changes in basal ganglia oxidative metabolism and motor response to apomorphine induced by nigrostriatal lesion in rats.

In Parkinson's disease, the functional architecture of the basal ganglia nuclei undergoes profound alterations, one of the most important of which is overactivity of the basal ganglia output nuclei. This phenomenon seems to be intimately related to pathological overactivity of the subthalamic nucleus, which directly modulates the basal ganglia output through its glutamatergic projections. In this study, we investigated the effects of unilateral subthalamic nucleus lesions on the activities of succinate dehydrogenase and cytochrome oxidase, two markers of neuronal activity, in rats with prior unilateral lesions of the nigrostriatal tract. We also explored the effect of subthalamic nucleus lesions on the rotational response to systemic apomorphine. Rats with unilateral lesions of the nigrostriatal tract showed ipsilateral increases in enzyme activity in the basal ganglia output nuclei, entopeduncular nucleus and substantia nigra pars reticulata. Selective subthalamic nucleus destruction completely reversed this phenomenon. In addition, subthalamic nucleus lesions abolished the rotational response to apomorphine. These results confirm that overactivity of the subthalamic nucleus plays a pivotal role in the functional alterations of basal ganglia associated with Parkinson's disease. They also shed further light on the neural mechanisms through which manipulations of subthalamic activity can ameliorate Parkinson's disease symptoms.

Effects of sodium concentration on human neutrophil bactericidal functions

What are the ionic requirements for neutrophil (PMN) function and how might altered electrolyte concentrations contribute to airway disease? The in vitro killing of Pseudomonas aeruginosa by human peripheral white blood cells (WBCs) was progressively compromised Na+ concentration was lowered from 124 to 62 mM; at 62 mM Na+, bactericidal activity was 28.8 +/- 7.4% (SE) of normal. In contrast, Cl- concentration affected killing only when lowered to 8 mM. We examined phagocytosis and oxidative metabolism in response to P. aeruginosa or particles opsonized with either immunoglobulin G (IgG) or complement (C'). Phagocytosis of P. aeruginosa and of IgG-coated particles was Na(+)-dependent (31.2 +/- 3.1 and 58.6 +/- 14.2% of normal, respectively, at 62 mM Na+). However, no effect on uptake of C'-coated particles was observed, and the respiratory burst at 70 mM Na+ was normal regardless of stimuli. Thus low Na+ concentration compromises select PMN functions. These results may help explain why airways of cystic fibrosis (CF) patients become colonized with bacteria such as P. aeruginosa. Perhaps the low concentration of Na+ reported for some CF respiratory secretions inhibits bactericidal functions of PMNs, predisposing these patients to airway infections.

Distinguishing features of basophil and neutrophil activation by major basic protein

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An antibody that binds a neutrophil membrane protein, ERp72, primes human neutrophils for enhanced oxidative metabolism in response to formyl-methionyl-leucyl-phenylalanine. Implications for ERp72 in the signal transduction pathway for neutrophil priming.

Human neutrophils are primed by cytokines for enhanced oxidative metabolism in response to chemotactic factors, but the signal transduction pathways for cytokine activation and priming are unknown. Neutrophil priming may play an important role in mechanisms of host defense and inflammatory responses associated with autoimmune diseases. A rabbit antibody was produced that reacted with human neutrophils and induced priming in response to the chemoattractant, FMLP. The protein responsible for neutrophil priming in response to binding antibody was identified in a neutrophil cDNA library by expression cloning. The cloned protein absorbed the neutrophil-priming activity from rabbit serum. Furthermore, antibody priming activity was recovered by elution from the cloned protein. The gene for the protein associated with neutrophil priming was sequenced and identified as the endoplasmic reticulum protein, ERp72, which contains three copies of the active site sequences of protein disulfide isomerase. The antibody that primed neutrophils was shown to bind ERp72 in neutrophil membranes by immunoprecipitation of the same 72-kDa protein from neutrophils as a known antibody to ERp72. These studies implicate ERp72 in the signal transduction pathway for priming human neutrophils.

Virulence factors of the family Legionellaceae.

Whereas bacteria in the genus Legionella have emerged as relatively frequent causes of pneumonia, the mechanisms underlying their pathogenicity are obscure. The legionellae are facultative intracellular pathogens which multiply within the phagosome of mononuclear phagocytes and are not killed efficiently by polymorphonuclear leukocytes. The functional defects that might permit the intracellular survival of the legionellae have remained an enigma until recently. Phagosome-lysosome fusion is inhibited by a single strain (Philadelphia 1) of Legionella pneumophila serogroup 1, but not by other strains of L. pneumophila or other species. It has been found that following the ingestion of Legionella organisms, the subsequent activation of neutrophils and monocytes in response to both soluble and particulate stimuli is profoundly impaired and the bactericidal activity of these cells is attenuated, suggesting that Legionella bacterial cell-associated factors have an inhibitory effect on phagocyte activation. Two factors elaborated by the legionellae which inhibit phagocyte activation have been described. First, the Legionella (cyto)toxin blocks neutrophil oxidative metabolism in response to various agonists by an unknown mechanism. Second, L. micdadei bacterial cells contain a phosphatase which blocks superoxide anion production by stimulated neutrophils. The Legionella phosphatase disrupts the formation of critical intracellular second messengers in neutrophils. In addition to the toxin and phosphatase, several other moieties that may serve as virulence factors by promoting cell invasion or intracellular survival and multiplication are elaborated by the legionellae. Molecular biological studies show that a cell surface protein named Mip is necessary for the efficient invasion of monocytes. A possible role for a Legionella phospholipase C as a virulence factor is still largely theoretical. L. micdadei contains an unusual protein kinase which catalyzes the phosphorylation of eukaryotic substrates, including phosphatidylinositol and tubulin. Since the phosphorylation of either phosphatidylinositol or tubulin might compromise phagocyte activation and bactericidal functions, this enzyme may well be a virulence factor. Administration of the L. pneumophila exoprotease induces lesions resembling those of Legionella pneumonia and kills guinea pigs, suggesting that this protein plays a role in the pathogenesis of legionellosis. However, recent work with a genetically engineered strain has convincingly shown that the protease is not necessary for intracellular survival or virulence. As might be expected with a complex process like intracellular parasitism, it appears that the capability of Legionella strains to invade and multiply in host phagocytes is multifactorial and that no single moiety which is responsible for the virulence phenotype will be found.

Virulence factors of the family Legionellaceae.

Whereas bacteria in the genus Legionella have emerged as relatively frequent causes of pneumonia, the mechanisms underlying their pathogenicity are obscure. The legionellae are facultative intracellular pathogens which multiply within the phagosome of mononuclear phagocytes and are not killed efficiently by polymorphonuclear leukocytes. The functional defects that might permit the intracellular survival of the legionellae have remained an enigma until recently. Phagosome-lysosome fusion is inhibited by a single strain (Philadelphia 1) of Legionella pneumophila serogroup 1, but not by other strains of L. pneumophila or other species. It has been found that following the ingestion of Legionella organisms, the subsequent activation of neutrophils and monocytes in response to both soluble and particulate stimuli is profoundly impaired and the bactericidal activity of these cells is attenuated, suggesting that Legionella bacterial cell-associated factors have an inhibitory effect on phagocyte activation. Two factors elaborated by the legionellae which inhibit phagocyte activation have been described. First, the Legionella (cyto)toxin blocks neutrophil oxidative metabolism in response to various agonists by an unknown mechanism. Second, L. micdadei bacterial cells contain a phosphatase which blocks superoxide anion production by stimulated neutrophils. The Legionella phosphatase disrupts the formation of critical intracellular second messengers in neutrophils. In addition to the toxin and phosphatase, several other moieties that may serve as virulence factors by promoting cell invasion or intracellular survival and multiplication are elaborated by the legionellae. Molecular biological studies show that a cell surface protein named Mip is necessary for the efficient invasion of monocytes. A possible role for a Legionella phospholipase C as a virulence factor is still largely theoretical. L. micdadei contains an unusual protein kinase which catalyzes the phosphorylation of eukaryotic substrates, including phosphatidylinositol and tubulin. Since the phosphorylation of either phosphatidylinositol or tubulin might compromise phagocyte activation and bactericidal functions, this enzyme may well be a virulence factor. Administration of the L. pneumophila exoprotease induces lesions resembling those of Legionella pneumonia and kills guinea pigs, suggesting that this protein plays a role in the pathogenesis of legionellosis. However, recent work with a genetically engineered strain has convincingly shown that the protease is not necessary for intracellular survival or virulence. As might be expected with a complex process like intracellular parasitism, it appears that the capability of Legionella strains to invade and multiply in host phagocytes is multifactorial and that no single moiety which is responsible for the virulence phenotype will be found.