Species distribution and susceptibility to azoles of vaginal yeasts isolated prostitutes.

OBJECTIVE: We investigated the use of miconazole among female prostitutes in Costa Rica as well as the distribution of vaginal yeasts and the susceptibility pattern to azoles of strains obtained from this population. Our intention was to relate a frequent use of miconazole to occurrence of vaginal yeasts resistant to azoles. METHODS: Vaginal samples were taken from 277 patients that have previously used azoles. Vaginal swabs were obtained for direct microscopy and culture. Yeast isolates were identified by germ tube test and assimilation pattern. Susceptibility testing was determined using a tablet diffusion method. RESULTS: The number of clinical Candida isolates (one from each patient) was 57 (20.6%). C. albicans was the predominant species (70%), followed by C. parapsilosis (12%), C. tropicalis (5.3%), C. glabrata and C. famata (3.5% each), C. krusei, C. inconspicua and C. guilliermondii (1.7% each). The majority of vaginal Candida isolates were susceptible to ketoconazole (91%), fluconazole (96.5%), and itraconazole (98%). A lower susceptibility of some isolates to miconazole (63%) was observed as compared to the other azoles tested. Moreover, the strains, nonsusceptible to miconazole, were more often obtained from patients that have used this antifungal at least four times within the last year before taking the samples as compared to those with three or less treatments (P<.01). CONCLUSION: An indiscriminate use of miconazole, such as that observed among female prostitutes in Costa Rica, results in a reduced susceptibility of vaginal yeasts to miconazole but not to other azoles.

The effect of intralipid on mononuclear and polymorphonuclear phagocytes.

Eight healthy subjects were given Intralipid, a soybean oil emulsion, 20% intravenously for 2 h. During the infusion a significant increase in the nitroblue tetrazolium-reduction of blood monocytes was noted. Preincubation of monocytes in vitro with Intralipid (20 to 100 mg/ml) for 30 min was found to increase the ability of the cells to migrate chemotactically and to phagocytize yeast particles. On the contrary, when neutrophilic granulocytes were preincubated with Intralipid in the same concentrations for 30 min. their nitroblue-tetrazolium-reduction, chemotactic and spontaneous locomotion, as well as their ingestion of yeast particles was depressed.

Decreased chemotactic and random migration of leukocytes during Intralipid infusion.

The soybean oil emulsion Intralipid was given intravenously to 12 healthy subjects for 2 hr. During the infusion an impairment of the chemotactic and random migration of leukocytes was noted. It was correlated to the dose given and to the degree of hypertriglyceridemia induced. Migration was fully restituted 22 hr after the infusion. Also when added in vitro Intralipid caused an impairment of leukocyte motility that followed a dose response pattern.

Alveolar macrophages and lung lesions after combined exposure to nickel, cobalt, and trivalent chromium.

In earlier inhalation exposures of rabbits, nickel increased the production of surfactant by type II cells, with secondary effects on morphology and function of alveolar macrophages. Cobalt induced mainly a nodular growth pattern of the type II cells. Trivalent chromium seemed to impair the capacity of macrophages to catabolize surfactant but did not affect the type II cells. We exposed rabbits by inhalation to combinations of nickel (0.6 mg/m3 as NiCl2) and trivalent chromium [1.2 mg/m3 as Cr(NO3)3] (Ni-Cr), cobalt (0.5 mg/m3 as CoCl2) and nickel (0.5 mg/m3) (Co-Ni), or cobalt (0.5 mg/m3) and chromium (1.2 mg/m3) (Co-Cr) for 4 months, 5 days/week, 6 hr/day. Alveolar macrophages, alveolar type II cells, and lung content of phospholipids were determined. All combined exposures induced more pronounced lung lesions than exposures for each of the metals. Phospholipid concentrations were significantly higher. There were significantly higher percentages of macrophages filled with surfactant-like inclusions and a smooth surface. Accumulations of macrophages in alveoli were more widespread. Chromium potentiated the effects of nickel and cobalt on the type II cells, which led to secondary effects on the macrophages. Nickel potentiated the specific effects of cobalt, i.e., type II cell nodule formation. The result indicates that noxious effects could also be induced in man by combined exposure to nickel, cobalt, and trivalent chromium in concentrations similar to those occurring in some occupational settings.

Fungal prophylaxis by reduction of fungal colonization by oral administration of bovine anti-Candida antibodies in bone marrow transplant recipients.

Candida overgrowth and invasion constitute a serious threat with a high mortality in BMT recipients. Currently available topical antifungal prophylaxis is largely ineffective, and as resistance to existing, absorbable drugs for systemic use is rapidly developing, new forms of therapy are needed. We investigated the effect of oral treatment of BMT recipients with a bovine immunoglobulin product derived from animals immunized against several Candida species. The natural Candida colonization was first followed in 19 patients to establish the colonization pattern. Half of the patients were found to be colonized prior to transplantation and altogether 72% were colonized at some point during follow-up. Those with a high pre-transplant concentration of Candida in saliva (>100 CFU/ml) remained colonized throughout the BMT treatment period. The therapeutic effect was monitored in two other patient groups. The first group consisted of nine patients, where, due to a low number of primary colonized patients, response in colonized patients was suggestive of a therapeutic effect. In the second group, 10 patients with a high level of colonization (>100 CFU/ml) were given 10 g daily of the product in three divided doses. The results suggest a treatment-related reduction in Candida colonization in a majority (7/10) of patients and one patient became completely negative. As no adverse effects were noted, our findings encourage additional studies in immunocompromised, transplant patients.

Migration of Entamoeba histolytica under agarose.

A procedure for visualizing and quantifying motility of Entamoeba histolytica by migration under agarose is described. Agarose suspended in tissue culture medium 199 supplemented with bovine albumin was poured into plastic dishes and allowed to harden. Six pairs of wells were cut out in a circular configuration. To the inner wells a suspension of E. histolytica in Eagle's Medium (24 X 10(6) cells/ml) was added, and to the outer wells a chemoattractant or the control medium. After overnight incubation at 37 degrees C, the amebae were fixed and stained. The chemotactic and spontaneous migrations were measured in an enlarging projector. Escherichia coli filtrates, suspensions of intact and lysed erythrocytes, and the complement factor C5a acted as good chemoattractants. Both the random and chemotactic motility were correlated to the time of the incubation. Cytochalasin B effected a dose-related inhibition of both chemotactic and random migration, while colchicine caused a decrease of the chemotaxis only. The reproducibility of the method, measured by 10 intra-assay tests, was good. Thus, the described method can be useful for comparative determinations of the motility of different ameba populations. Furthermore, different factors affecting the motility of amebae can be studied.

The effect of phospholipid-containing surfactant from nickel exposed rabbits on pulmonary macrophages in vitro.

Alveolar macrophages from 9 normal rabbits were incubated in vitro for 3 h with and without phospholipid-containing surfactant from nickel-treated ones. The macrophages treated with surfactant showed morphological and functional criteria of increased activity. The cell surface had many protrusions and the cytoplasma contained several lamellated structures. The oxidative metabolism, measured by the nitroblue tetrazolium (NBT)-test, at rest and after E. coli stimulation was increased, as was the attachment and ingestion of yeast particles. The NBT-values were about the same as corresponding values of macrophages lavaged from the lungs of nickel-treated rabbit. Macrophages incubated with surfactant from untreated animals, had NBT values and phagocytic activity similar to cells incubated without surfactant. As this substance was administered in excess, the difference in macrophage response would probably be due to a qualitative alteration of the surfactant after nickel exposure.

Alveolar macrophage function in nickel dust exposed rabbits.

8 rabbits were exposed to metallic nickel dust (2 mg/m3, of which about half was respirable) for 4 weeks. The lungs were lavaged and the macrophages were collected. In comparison with 8 control rabbits, a significant increase was noted in the nickel exposed rabbits as concerned the weight and density of the lungs, the size variation of the lung cells, the phagocytosis of silver coated particles, and the metabolic activity as measured by NBT reduction. The last mentioned increase was recorded during basal conditions as well as during phagocytosis. The NBT reduction during phagocytosis was significantly correlated with the degree of phagocytosis of silver coated particles in both control and exposed rabbits. It is suggested that the exposure to nickel dust has unspecifically activated the macrophages perhaps by increased production of phospholipids.

Clinical tolerance and metabolism of a fat emulsion containing carnitine and gamma-linolenic acid.

The purpose of the present study was to study the metabolism and evaluate the tolerance of a new fat emulsion, which is rich in gamma-linolenic acid and has been supplemented with carnitine. 24 adult male volunteers participated in an open, randomised, crossover study where half of the subjects were given Intralipid 20% (IL) on day one followed by administration of the supplemented fat emulsion (FE) on day two, the other half being treated in the reverse order. Both fat emulsions were first administered as a bolus dose of 0.10 g triglycerides per kg BW (an intravenous fat tolerance test) followed 40 min later by a continuous intravenous infusion during 4 h at a rate of 0.21 g triglycerides per min. The plasma fractional elimination rate, k(2)-value, was lower for FE than for IL (4.6 +/- 1.8 vs 5.3 +/- 1.6% per min, P < 0.05, mean +/- SD). Infusion of FE, as distinct from IL administration, was accompanied by a marked increase in serum gamma-linolenic acid concentration (239 +/- 133 vs 0 +/- 11 mumol/l, P < 0.05). No differences between FE and IL were discernible regarding the serum concentrations of dihomo-gamma linolenic acid (C 20: 3omega6), arachidonic acid (C 20: 4omega6), adrenic acid (C 22: 4omega6) or docosapentanaenoic acid (C 22: 5omega6). Leucocyte chemotaxis and random migration increased, 0.5 +/- 0.3 arb. units (P < 0.01) and 0.2 +/- 0.2 arb. units (P < 0.05), during infusion of FE but no such effect was noted during IL administration. No effect of the fat emulsion composition and carnitine was detected on phagocytosis and oxidative metabolism of neutrophil leucocytes. In conclusion, the new fat emulsion containing a relatively high proportion of gammalinolenic acid and supplemented with carnitine was found to be well tolerated although it was eliminated from plasma at a 13% slower rate. The supplemented fat emulsion had a moderate stimulatory effect on leukocyte chemotaxis.

Lymphocyte and granulocyte function in nutritionally depleted patients. The effect of 2 weeks of total parenteral nutrition.

Parenteral nutrition has been reported to restore impaired immune function in nutritionally depleted patients. Lymphocyte and granulocyte function were studied before and after 2 weeks of total parenteral nutrition (TPN) in 9 depleted patients, 5 with gastrointestinal cancer and 4 with benign gastrointestinal disorders. The ability of purified blood lymphocytes to incorporate (14)C-labelled thymidine spontaneously and following mitogenic and antigenic stimulation was studied. Before TPN spontaneous lymphocyte activity was significantly increased and mitogen and antigen responsiveness significantly decreased in comparison to healthy controls. These abnormalities were more pronounced in patients with malignant disorders while the remaining patients showed a moderate impairment. No improvement in lymphocyte capacity was observed following 2 weeks of TPN. Granulocyte function measured as chemotaxis, phagocytosis, NBT-reduction and bactericidal capacity did not differ significantly between patients and controls and no changes were seen following nutritional treatment. These findings support the concept that the underlying disease and/or other factors rather than the secondary nutritional depletion are the main reasons for immune impairment. Furthermore, no immunomodulating effects of lipid emulsions on lymphocyte and granulocyte function, under these experimental conditions, were apparent.

Plasma clearance of intravenous fat emulsion in acute bacterial infections.

A variety of abnormalities in lipid metabolism during infection have been reported and it has been questioned whether intravenous fat emulsion should be given in septic states. This study was designed to investigate the removal of a fat emulsion from plasma during septicaemia. An intravenous fat tolerance test (0.1 g of fat in the form of Intralipid 10% per kg b.w.) was performed in 28 adult patients who had been admitted for suspected septicaemia. All patients were febrile (mean body temperature 38.9 +/- 0.1 degrees C SEM), the majority had leucocytosis (mean value 10.5 +/- 1.0 10 (9)1 ) and all were haemodynamically stable. On the basis of bacterial cultures taken from various locations, the patients were classified as septic (n = 12; gram-negative sepsis, n = 8; gram-positive sepsis, n = 4), infected but without septicaemia (n = 9) or febrile of unknown cause (all cultures negative, n = 7). Plasma fractional removal rates of intravenous fat were similar in the different groups. The mean plasma clearance rate of intravenous fat was 6.6 +/- 0.5 (% minute), which does not differ from that reported for healthy controls. All patients with proven septicaemia and 5 non-septic patients were studied two weeks later when their infection had been treated and they were off antibiotic medication. The plasma clearance rate of i.v. fat was significantly lower in the recovery phase than during the acute febrile state (5.9 +/- 0.5 vs 4.2 +/- 0.4 %/min, n = 17; p < 0.05). It is concluded that clearance of intravenous fat emulsions from plasma is unimpaired in haemodynamically stable patients with septicaemia or other acute infections.

Interaction between lung surfactant and nitric oxide production by alveolar macrophages stimulated by group B streptococci.

The major etiologic agent in neonatal pneumonia and meningitis is group B streptococci (GBS). Nitric oxide (NO) production by alveolar macrophages (AM) in response to Gram-positive bacteria such as GBS and the effect of surfactant on this production have received little attention. We studied production of NO by GBS-stimulated AM using the Griess reaction, the effect of lung surfactant on this NO production, and the possible lipid peroxidation (LPO) of surfactant caused by NO. The LPO test was used to measure surfactant peroxidation. Heat-killed and live GBS were found to stimulate NO production by rat alveolar macrophages, and the presence of interferon gamma (IFN-gamma) increased this stimulation in a synergistic manner. Curosurf(R), the natural surfactant used in our study, significantly reduced NO production in various sets of experiments. Lipid peroxidation of surfactant was noted when NO was produced by stimulated AM, a phenomenon that could be suppressed by NG-monomethyl L-arginine (L-NMMA), the inhibitor of NO synthase. In the lung of GBS-infected neonates, nitric oxide produced by AM might contribute to the destruction of surfactant caused by inflammatory cells. Pediatr Pulmonol. 2000; 30:106- 113.

Phagocyte-induced lipid peroxidation of lung surfactant.

Surfactant therapy is given routinely to premature newborns with respiratory failure. However, alterations in surfactants have been shown to be a significant factor in some forms of respiratory failure in newborns in animal models of lung injury. To investigate whether antioxidant supplementation might help to protect exogenous surfactant from damage by oxygen free radicals, we examined the influence of vitamin E in combination with surfactant on superoxide production as estimated by the nitroblue tetrazolium reduction test, and measured surfactant peroxidation with a new colorimetric method with or without addition of superoxide dismutase (SOD) or vitamin E. Our results showed that surfactant interacts with free radicals; surfactant reduced superoxide production by neutrophils and was peroxidized when incubated with resting and with stimulated cells. Vitamin E supplementation decreased superoxide radical production and in a dose-dependent manner decreased surfactant peroxidation. The decrease in lipid peroxidation by SOD was not significant. These findings suggest that phagocytes induce lipid peroxidation of lung surfactant, a reaction that might be prevented by antioxidants.

Increased reduction of nitroblue tetrazolium by human blood monocytes following post-operative radiation therapy for breast cancer.

Reduction of nitroblue tetrazolium by blood monocytes was significantly increased following post-operative radiation therapy for breast cancer. Attachment and ingestion of yeast particles by monocytes was not affected. Exposure of purified monocytes in vitro to 13 Gy did not significantly affect NBT reduction of yeast particle attachment, whereas ingestion of yeast particles was slightly increased.

Neutrophil function and glutathione-peroxidase (GSH-px) activity in healthy individuals after treatment with N-acetyl-L-cysteine.

The objective of this study was to evaluate the effect of N-acetyl-L-cysteine (NAC) on neutrophilic functions and as an antioxidant. NAC, 600 mg daily, given orally to healthy individuals for a period of 2 weeks, affected some functions of human neutrophilic granulocytes when tested in vitro. NAC treatment caused a decrease in the production of superoxide anions by stimulated neutrophils and the improvement of their phagocytic capacity although it did not affect their random or chemotactic migration. The level of glutathione peroxidase (GSH-px) in thrombocytes of the NAC-treated individuals was increased in comparison with the activity before treatment. These results suggest that NAC might act as a scavenger of oxygen-derived free radicals released by stimulated neutrophils and thereby protect the tissue against the radical caused injury as well as optimize phagocytosis.

Oxygen radical release by neutrophils of HIV-infected patients.

Neutrophils from asymptomatic HIV-infected patients have an increased Nitroblue tetrazolium (NBT) reduction, that is an increased production of oxygen radicals. Plasma from these patients can activate normal neutrophils to an increased NBT-reduction and the neutrophil activating factor thus seems to be mainly plasma bound. Further, the patients also have increased levels of plasma malondialdehyde and thus an increased lipid peroxidation. Plasma cysteine levels are low, a sign of increased consumption of antioxidants. Treatment of the asymptomatic HIV-infected patients with N-acetylcysteine corrected the plasma cysteine levels and had some beneficial effects, but did not inhibit the increased radical production by the neutrophils.

Surfactant improves lung function and mitigates bacterial growth in immature ventilated rabbits with experimentally induced neonatal group B streptococcal pneumonia.

AIMS: To study the influence of surfactant on lung function and bacterial proliferation in immature newborn rabbits with experimental group B streptococcal (GBS) pneumonia. METHODS: Preterm rabbit fetuses (gestational age 28 days) underwent tracheotomy and were mechanically ventilated in a warmed body plethysmograph that permitted measurement of lung-thorax compliance. Fifteen minutes after the onset of ventilation the animals received either GBS or saline intratracheally; at 30 minutes, a bolus of saline or 200 mg/kg of a porcine surfactant (Curosurf) was administered via the airway. Bacterial proliferation was evaluated in lung homogenate at the end of the experiments and the results expressed as mean log10 cfu/g lung (SD). Animals receiving only saline (n = 20) or saline and surfactant (n = 20) served as controls. RESULTS: The average survival time was about three hours in all groups. Infected animals receiving surfactant (n = 22) had significantly less bacterial growth (9.09 (0.45) vs 9.76 (0.91)) and improved lung function (compliance: 0.61 (0.14) vs 0.34 (0.19) ml/kg. cm H2O) than infected rabbits receiving saline at 30 minutes (n = 22). CONCLUSION: Surfactant improves lung function and mitigates bacterial growth in preterm rabbits infected with group B streptococci.

Phagocyte-induced lipid peroxidation of different intravenous fat emulsions and counteractive effect of vitamin E.

Unsaturated fatty acids, a major component of fat emulsions used in parenteral nutrition, are prone to peroxidation which is an important feature of oxygen-associated tissue damage. We used the nitroblue tetrazolium (NBT) reduction test to measure the production of superoxide radicals by stimulated polymorphonuclear neutrophils (PMN) in the presence of different fat emulsions: Intralipid (containing 100% long-chain triacylglycerols, LCT), Vasolipid (a physical mixture of 50% LCT and 50% medium-chain triacylglycerols, MCT) and Structolipid (structured triacylglycerols containing 63% LCT and 37% MCT). We measured the amount of malonaldehyde (MDA) and 4-hydroxyalkenal to determine the lipid peroxidation of the three fat emulsions in the presence of stimulated neutrophils. Further, we investigated the role of vitamin E (alpha-tocopherol) in preventing lipid peroxidation in vitro. The results showed that the values of NBT reduction of PMN were significantly decreased in each of the three fat emulsions and that increasing concentrations of fat emulsions were associated with decreased values of NBT reductions, in a dose-dependent way (P<0.001). There were, however, no statistically significant differences between the values of the three different types of fat emulsions (P>0.05). Lipid peroxidation increased significantly in the presence of all three types of fat emulsions, and was more pronounced for Intralipid than for Vasolipid and Structolipid after 1 and 2 h of incubation with resting as well as with stimulated phagocytes. The increased lipid peroxidation of the fat emulsions was markedly reduced by vitamin E, and the inhibition was concentration dependent. In conclusion, lipid peroxidation in vitro is more pronounced when PMNs are incubated with fat emulsions. This increase in lipid peroxidation can be reduced by adding vitamin E to the fat emulsions.

Effect of surfactant and specific antibody on bacterial proliferation and lung function in experimental pneumococcal pneumonia.

OBJECTIVE: To investigate the effect of surfactant and specific antibody on bacterial proliferation in experimental pneumococcal pneumonia. METHODS: Near-term newborn rabbits received a standard dose (10(7)) of type 3 pneumococci via the airways. Control animals were sacrificed 1 minute later. Other animals were ventilated for 5 hours and treated via the tracheal cannula with surfactant (Curosurf 200 mg/kg), a mixture of surfactant and a polyclonal antipneumococcal antibody, the antibody without surfactant, or saline. RESULTS: There was a significant bacterial proliferation in lung tissue in all animals ventilated for 5 hours. Bacterial growth, expressed as log10 colony forming units (CFU) per gram of lung tissue was less prominent in animals treated with a mixture of surfactant and specific antibody than in animals treated with antibody alone (median, 7.51, range, 6.80--7.70 vs. median, 7.92, range, 7.07--8.50; P < 0.05). Dynamic lung-thorax compliance was improved with surfactant or surfactant plus antibody in comparison with saline or antibody alone. CONCLUSIONS: The data suggest that the suppressive effect of the antibody on bacterial proliferation becomes evident only when surfactant is administered together with the antibody.

Human granulocyte and reticuloendothelial system function during intralipid infusion.

In six patients, the phagocytic and catabolic functions of the RES (reticuloendothelial system), as measured by using 125I-microaggregated human serum albumin as a test substance, were not affected by Intralipid infusion. The nitroblue tetrazolium (NBT) reduction of granulocytes, the NBT reduction and the bacplasma, was significantly lower in eight patients during Intralipid infusion than before this treatment. When heat-killed E. coli were added for phagocytic stimulation in vitro, the same result was obtained only in the absence of plasma. The bactericidal capacity of granulocytes was significantly decreased in six patients during Intralipid infusion. When Intralipid was added in vitro to granulocytes, the NBT reduction, and the bactericidal capacity of these cells were decreased and the effect was mainly dose-dependent. Electron micrographs show that granulocytes phagocytize Intralipid. The loading of granulocytes with lipid particles may block the engulfment of bacteria and may be a reason for the decreased bactericidal capacity of these cells. An influence exerted by the lipid on the cell membrane is the most conceivable reason for the decreased NBT reduction.