The method for estimation of opsonic activity in sera of infants

Three methods for the estimation of opsonic activity in the sera of newborn children were tested. Two of them, based on the phagocytosis of opsonised bacteria labelled with radioactive phosphorus³²P as measured byin vivo blood clearance or uptake of bacteria in perfused isolated liver, were found to be unsuitable for long term dynamic study mainly because they do not permit the testing of series of samples. The third method (using isolated phagocytic cellsin vitro) permits the differentiation of the opsonic effect of complement and antibody and, furthermore, the firmness of the bond between microbes and phagocytes (which reflects the degree of opsonization) can be established. It was found that a 2-mercaptoethanol-resistant antibody, probably of the IgG type, was responsible for the opsonic activity of children's sera toEscherichia coli 083. Homologous antibody (toEscherichia coli 083) could be differentiated from beterologous antibody (toEscherichia coli 086) using the opsonic test only at low dilutions of sera. The combination of newborn piglet complement and antibody of children's sera yielded higher values of opsonic activity than either component separately.     

Development of pulmonary intravascular macrophage function in newborn lambs.

We sought to determine whether pulmonary intravascular macrophages are involved in pulmonary vascular sensitivity to intravenously injected particles in sheep. We estimated that newborn lambs have few of these macrophages at birth but develop a 10-fold greater density within 2 wk. Awake, chronically instrumented newborn lambs showed no change in pulmonary vascular driving pressure (pulmonary arterial minus left atrial pressure) after injection of either liposomes [2 +/- 3 (SD) cmH2O; n = 5] or Monastral blue particles (3 +/- 2 cmH2O; n = 6) and showed no net pulmonary production of thromboxane B2, the stable metabolite of the vasoconstrictor thromboxane A2. In contrast, five of those lambs 2 wk later showed both an increase in pulmonary vascular driving pressure after injection of liposomes and Monastral blue (20 +/- 16 and 25 +/- 15 cmH2O, respectively; P < 0.05) and net pulmonary production of thromboxane B2 (171 +/- 103 and 429 +/- 419 pg/ml plasma, respectively; P < 0.05). Older lambs (n = 5) had higher pulmonary uptakes than newborn lambs (n = 6) of radioactive liposomes (47 +/- 13 vs. 12 +/- 10%; P < 0.01) and Monastral blue (53 +/- 6 vs. 21 +/- 10%; P < 0.05). We conclude that pulmonary intravascular macrophages are responsible for the sensitivity of sheep to intravenous foreign particles and are essential for a cascade of processes leading to microvascular injury.     

Pituitary somatotropic function of newborn infants]

The content of the hypophysial somatotrophic hormone was studied in the blood of cord vessels at parturition and in the peripheral blood of newborn infants during the first 2 hrs and 1-9 days of life. The hormone level by the periods of 2 hrs was markedly lower than in the foetuses, it sharply increased by the end of the first day and gradually decreased during the subsequent days. The functioning of the hypothalamic-hypophysial system in humans during this period is discussed.     

The opsonic activity of complement to rough strains ofEscherichia coli in vivo in newborn precolostral pigs

The participation of complement of newborn precolostral piglets (in the absence of antibody) in the opsonization of roughEscherichia coli was investigated using blood clearance test. It was found that the complement inhibitor-yeast mannan inhibits thein vivo blood clearance when injected i. venously into newborn precolostral piglets. This inhibition takes place at the serum level and not by saturation of RES cells by mannan: sera from mannan-treated piglets are not bactericidal in anin vitro bactericidal assay against roughEscherichia coli; secondly, the presence of antibody removes the blockade of phagocytosis by mannan indicating that the RES cells are functionally capable to perform phagocytosis. On the basis of our findings we postulate that in newborn precolostral piglets in which antibody is not present in their sera, complement alone (or with cooperation with other unknown serum component) acts as an opsonin on roughEscherichia coli and is responsible for the blood clearance rate of these bacteria.     

Effects of 2.5‐hour sumo training on serum opsonic activity

Sumo is a traditional Japanese sport, but the effect of actual daily training on neutrophil function is unknown. We evaluated the effect of sumo training on serum opsonic activity (SOA), which is one of the main neutrophil‐related functions. Seventeen male university sumo wrestlers participated in the study. Changes in anthropometric parameters, concentrations of serum immunoglobulins (IgG, IgA and IgM), complements (C3 and C4), myogenic enzymes (lactate dehydrogenase, asparate aminotransferase, alanine aminotransferase and creatine kinase), white blood cell/neutrophil counts and SOA were measured immediately before and after actual daily training for 2.5 h. Compared with the pre‐values, immunoglobulins and complements, myogenic enzymes and white blood cell/neutrophil counts significantly increased (p <0.01 for all). As for SOA, the values of the peak height and the area under the curve significantly increased after the training when assessed using lucigenin as a chemiluminigenic probe (p <0.01 for all), but showed no significant change when luminol was used as the chemiluminigenic probe. In conclusion, daily actual sumo training for 2.5 h increases SOA, thus possibly activating the reactive oxygen species production of neutrophils. Copyright © 2009 John Wiley & Sons, Ltd.     

Surfactant therapy of newborn rabbits impairs lung macrophage bactericidal activity

Because in vitro studies indicate that pulmonary alveolar macrophages (PAM's) filled with phospholipid vesicles have depressed microbicidal capacity, we tested the intrapulmonary bactericidal activity of newborn PAM's after surfactant treatment. Term newborn rabbits received intratracheally either homologous surfactant or one of two artificial phospholipid vesicle preparations followed by pulmonary aerosol infection with group B streptococci (GBS). Four hours after lung infection, phagocytic killing of GBS was reduced by 70-90% in animals treated with the homologous and one of the artificial surfactants compared with untreated animals or animals that received intrapulmonary injections of the surfactant vehicle (P less than 0.02). The other artificial phospholipid preparation decreased intrapulmonary inactivation of GBS by 30-40% compared with the controls. The phospholipid vesicles in the three preparations were avidly ingested and processed by newborn PAM's. The diminished in vivo killing of GBS was not attributed to decreased viability or phagocytic behavior of the PAM's toward GBS. The bactericidal defect that was evident in the newborn PAM's appeared related to the uptake of large phospholipid vesicles in the preparations rather than to the phospholipid content of the surfactants themselves. When in vitro conditions that stimulated the alveolar environment were used, the natural surfactant preparation promoted GBS proliferation, whereas the artificial preparations did not. Our findings indicate that surfactant administration reduces the bactericidal activity of neonatal PAM's. We conclude that additional investigations are needed to ascertain the effect of surfactant replacement therapy on lost defenses of the lung.     

Measuring equipment for bedside diagnosis of respiratory function in spontaneously breathing newborn infants

Equipment was developed for bedside lung function testing in the newborn using the simultaneous measurement of air flow rate, tidal volume, and esophageal pressure changes as a measure of transpulmonary pressure. The equipment has a number of advantages for the investigation of very low birthweight infants. A flow-through technique was used to eliminate the dead space of the face mask and a very thin micro-tipped catheter permits ready measurement of esophageal pressure. With this equipment, long-term measurements are also possible in oxygen-dependent newborns and the air-tightness of the mask can be monitored continuously. Long-time measurements in neonates are a prerequisite for standardizing the measuring conditions and adapting the duration of the measurement to the variability of the signals, especially in newborn with an irregular pattern of breathing.     

Effects of long‐distance running on serum opsonic activity measured by chemiluminescence

Exhaustive exercise such as long‐distance running has been shown to increase susceptibility to infection. In order to investigate whether serum opsonic activity plays a role in such conditions, we utilized luminol‐dependent and lucigenin‐dependent chemiluminescence (LmCL and LgCL). We took serum samples from 24 male marathon runners before and after running 30 km. Neutrophils were isolated from the peripheral blood of healthy volunteers. Serum opsonic activity was examined by measuring neutrophil ROS stimulated with zymosan particles opsonized by the serum samples. Immunoglobulin and complement levels in the serum were also measured. After a 30 km run, the maximum light emission was increased and the time to reach the maximum light emission was shortened significantly (p < 0.05) in LmCL. However, there were no significant changes in the immunoglobulin and complement levels. The increase of ROS production may suggest that serum opsonic activity is accelerated after running 30 km. Thus, serum opsonic activity might not play a significant role in the susceptibility to infection after long‐distance running. Copyright © 2003 John Wiley & Sons, Ltd.     

Reproductive experience modifies dopaminergic function, serum levels of prolactin, and macrophage activity in female rats

Reproductive experience (RE), i.e. pregnancy and lactation, induces physiological changes in mammals. Recent data show that neuroimmune interactions are modulated by a diversity of events involving neurotransmitters and neuropeptides. These molecules, particularly dopamine (DA), were reported to mediate the relevant cross talk between immune and neuroendocrine systems. Moreover, DA-mediated regulation of leukocyte function is a reasonable approach to investigate the DA-operated regulatory switch for immune-competent cells, such as macrophages. Therefore, the goals of the present study were to determine the effects of RE on: (1) dopaminergic function through hypothalamic levels of DA, dihydroxyphenylacetic acid (DOPAC), homovanilic acid (HVA), serotonin (5-HT), and 5-hydroxyindole acetic acid (5-HIAA); (2) basal levels of circulating prolactin (PRL); and (3) activity of peritoneal macrophage (phagocytosis and oxidative burst). A total of 16 adult (200-250 g) female Wistar rats were used, divided in two groups: nulliparous and primiparous. Approximately 2-3 weeks after weaning pups from the primiparous group, both groups of rats were tested. The findings indicate that: (1) DOPAC concentrations, DOPAC/DA and HVA+DOPAC/DA ratios decreased in primiparous rats as compared to virgin rats, (2) primiparous rats showed significantly lower serum PRL levels, and (3) phorbol miristate acetate (PMA)-induced oxidative burst was decreased in peritoneal macrophage from primiparous rats as compared to virgin rats. To test the possible positive correlation between serum levels of PRL and the intensity of oxidative burst by peritoneal macrophage, an extra experiment was done with adult virgin female rats treated with domperidone, an antagonist of DA receptors. Domperidone-treated animals showed increased serum levels of PRL and simultaneous increase in peritoneal macrophage oxidative burst. Thus, suggesting an indirect participation of hyperprolactinemia, induced by this treatment in peritoneal macrophage activity of female rats. These results suggest that a previous RE can modulate the activity of dopaminergic hypothalamic systems, while decreasing PRL serum levels and the oxidative burst of peritoneal macrophage. The neurochemical and hormonal RE-induced changes correlate with the immune alterations.     

Studies on the function of blood-liquor barrier in newborn infants]

Simultaneous measurement of the electrolytic conductivity of the cerebrospinal fluid and the blood serum in newborns has revealed a concentration gradient suggesting an active transfer of electrolytes into the liquor space. The extent of this concentration drop is regarded as expressing the functional capacity of the blood-liquor barrier. Comparative investigations in healthy older children shown that constant values, corresponding to the adult age, are reached approximately at the end of the first year of life.