Alveolar macrophage count as an indicator of lung reaction to industrial air pollution

The major proportion of the cellular components found in most of the sputa from persons working in polluted atmospheres are alveolar macrophages, and counts of the alveolar macrophages present in smears made from sputa from exposed workers probably reflect the lung reaction to air pollution. An investigation of this phenomenon was undertaken using the sputa of workers in different types of industries: a coke plant, two different aluminum reduction plants, a ship-building yard and asphalt work on a road. The results showed that the alveolar macrophage count increases with a higher level of particulate pollution in the workplace. A synergistic effect of occupational air pollution and smoking habits was also recorded.     

Alveolar macrophage response to experimentally induced ornithosis.

Effects of two strains of the agent of ornithosis upon alveolar macrophages of mice were compared. Macrophages were obtained by lavages of the lower respiratory tract. Experimental mice were intranasally infected with 100 LD50 of strain P 89 and strain Stanglová, respectively. Up to the culmination of the disease (from 6th to 8th day in P 89 and from 5th to 8th day in Stanglová), the number of harvested macrophages increased. The phagocytic index soon reached values around 40% (P 89) and 25% (Stanglová). Basophilic and eosinophilic macrophages increased in volume and their nuclear as well as plasma membranes became disrupted. A tendency of alveolar macrophages to fuse and form syncytial elements and sporadically, rosets, was observed. All cells in the lavage went through the same changes. Histological examination has shown initial changes in epithelial cells of bronchi and development of pneumonia after fusion of peribronchial leucocytic infiltrates. By employing the above described technique, no great changes in the quality of the effects of either strain P 89 or Stanglová were found. A slight difference was observed only in the degree of alveolar macrophage stimulation and in the onset of symptoms.     

Alveolar macrophage activation in experimental legionellosis.

Legionella pneumophila is a facultative intracellular parasite of alveolar macrophages. In vitro studies have shown that lymphokine-activated mononuclear phagocytes inhibit intracellular replication of L. pneumophila. To determine if recovery from legionellosis is associated with activation of alveolar macrophages in vivo to resist L. pneumophila, we studied an animal model of Legionnaires' disease. Rats were exposed to aerosolized L. pneumophila and alveolar macrophages were harvested during the recovery phase of infection. We compared these alveolar exudate macrophages with normal resident alveolar macrophages for the capacity to support or inhibit the intracellular growth of L. pneumophila. We also measured Ia expression as a marker of immunologic activation, and studied binding of bacteria, superoxide release, and the expression of transferrin receptors as potential mechanisms of resistance to L. pneumophila. For perspective on the specificity of these responses, we also studied alveolar exudate cells elicited by inhalation of heat-killed L. pneumophila, live Listeria monocytogenes, and live Escherichia coli. We found that alveolar exudate macrophages elicited by live L. pneumophila, but not heat-killed L. pneumophila, resisted the intracellular growth of L. pneumophila. Exudate macrophages in resolving legionellosis exhibited increased Ia expression, diminished superoxide production, and downregulation of transferrin receptors. Binding of L. pneumophila to exudate macrophages was indistinguishable from that to resident macrophages in the presence of normal serum, and augmented in the presence of immune serum. Alveolar exudate macrophages elicited by E. coli also inhibited growth of L. pneumophila, and exhibited a modest increase in Ia expression without change in transferrin receptors. Exudate cells induced by L. monocytogenes exhibited up-regulation of Ia without diminution of superoxide release. Alveolar cells harvested after inhalation of heat-killed L. pneumophila did not differ from resident alveolar macrophages in the expression of surface markers. These findings suggest that alveolar macrophages are immunologically activated in vivo to serve as effector cells in resolving legionellosis, and that live bacteria are required to induce this expression of immunity. The mechanism of resistance to parasitism by L. pneumophila may entail restriction of the intracellular availability of iron, but does not involve diminished bacterial binding or an augmented respiratory burst.     

Rare non-albicans candida species detected in different clinical diagnoses

In working on the incidence of yeasts we excluded from the tested set C. albicans, C. krusei, C. parapsilosis and C. tropicalis on the basis of morphological and biochemical properties. We found 16 rare species of yeasts: C. claussenii (16), C. guilliermondii (10), C. robusta (9), C. pulcherrima (8), C. zeylanoides (4), C. glabrata (3), C. lusitaniae (3), C. catenulata (2), C. mesenterica (2), C. utilis (2), C. freyschussii, C. intermedia, C. kefyr, C. lipolytica, C. mogii, C. pseudotropicalis (1 for each). These yeasts were detected mainly in cases of premature babies (22) from the nasopharynx (13), from the rectum (4), from the skin (23), from wound drains and from blood (1 for each), with gynaecological diagnoses (15) and rarely other diagnoses, such as malignancy (5), hypertension and respiratory infections (4 for each), kidney transplantation (3), dialysis, haemolytic-uremic syndrome (2 for each), perforation of gastric ulcus, otitis, prostatitis, ulcus cruris, coma, spina bifida, peritonitis and hepatic failure from different clinical material.     

Alveolar macrophages: functional heterogeneity within macrophage populations from rat lung

Alveolar macrophages were obtained from the unstimulated lungs of rats by repeated endobronchial lavage and an interstitial macrophage population prepared by mincing the lungs subsequent to the lavage process. Considerable heterogeneity was observed within these macrophage populations with respect to cell size, surface morphology and cytochemistry. Functional studies, involving assessment of IgG-Fc receptor density/avidity and the expression of cytostatic activity in cultures of mitogen-stimulated lymphocytes, revealed comparable heterogeneity, and further indicated a considerably higher degree of apparent stimulation in the alveolar versus the interstitial macrophage population. Parallel assessment of the functional activity of blood monocytes, the immediate precursors of these latter macrophages, indicated a lower state of activation again. This suggests that the lung interstitium normally provides an intermediate environment between the blood and the alveolar spaces, wherein blood monocytes may undergo maturational changes before their efflux into the alveoli.     

临床分离161株念珠菌菌种鉴定及氟康唑药敏试验分析

目的调查临床分离的念珠菌种类及其对氟康唑的敏感性。方法采用科玛嘉念珠菌显色培养基和YBC平板对山东大学齐鲁医院细菌室分离到的161株念珠菌进行鉴定,并对分离出的白念珠菌分别采用NCCLS推荐的微量稀释法和ROSCO药敏纸片法对氟康唑进行药物敏感性试验。结果在161株念珠菌中白念珠菌为69．57％,热带念珠菌为19．88％,近平滑念珠菌为4．97％,克柔念珠菌为2．48％,光滑念珠菌为1．86％,其他念珠菌为1．24％。两种方法药敏结果显示：112株白念珠菌对氟康唑的敏感率分别为96．43％和97．32％,仅有2株耐药。结论白念珠菌仍然是我院分离率最高的念珠菌,其次是热带念珠菌和近平滑念珠菌。白念珠菌对氟康唑仍敏感,耐药菌株极少。     

In vitro phagocytosis of several candida berkhout species by murine leukocytes

In vitro phagocytosis of thirteen Candida berkhout species by rat leukocytes was studied to assess a possible correlation between pathogenicity and phagocytosisYeast-leukocyte suspensions were mixed up for 3 h and phagocytic index, germ-tube formation and leukocyte candidacidal activity were evaluated. Highest values for phagocytosis were reached in all cases at the end of the first hour. Leukocyte candidacidal activity was absent. Only C. albicans produced germ-tubes.The various phagocytosis indices were determined depending on the Candida species assayed. Under these conditions, the more pathogenic species presented the lower indices of phagocytosis.It is determined that the in vitro phagocytic index may bear a close relationship with the pathogenicity of the Candida berkhout.     

Alveolar macrophage activation is a key initiation signal for acute lung ischemia-reperfusion injury

Lung ischemia-reperfusion (I/R) injury is a biphasic inflammatory process. Previous studies indicate that the later phase is neutrophil-dependent and that alveolar macrophages (AMs) likely contribute to the acute phase of lung I/R injury. However, the mechanism is unclear. AMs become activated and produce various cytokines and chemokines in many inflammatory responses, including transplantation. We hypothesize that AMs respond to I/R by producing key cytokines and chemokines and that depletion of AMs would reduce cytokine/chemokine expression and lung injury after I/R. To test this, using a buffer-perfused, isolated mouse lung model, we studied the impact of AM depletion by liposome-clodronate on I/R-induced lung dysfunction/injury and expression of cytokines/chemokines. I/R caused a significant increase in pulmonary artery pressure, wet-to-dry weight ratio, vascular permeability, tumor necrosis factor (TNF)-alpha, monocyte chemoattractant protein (MCP)-1, and macrophage inflammatory protein (MIP)-2 expression, as well as decreased pulmonary compliance, when compared with sham lungs. After AM depletion, the changes in each of these parameters between I/R and sham groups were significantly attenuated. Thus AM depletion protects the lungs from I/R-induced dysfunction and injury and significantly reduces cytokine/chemokine production. Protein expression of TNF-alpha and MCP-1 are positively correlated to I/R-induced lung injury, and AMs are a major producer/initiator of TNF-alpha, MCP-1, and MIP-2. We conclude that AMs are an essential player in the initiation of acute lung I/R injury.     

Chemotactic macrophage subpopulations defined by macrophage chemotactic factors from delayed hypersensitivity reaction sites

The chemotactic specificity of ia-positive and -negative macrophages was studied by using three macrophage chemotactic factors (MCF), -a, -b, and -c, isolated from delayed hypersensitivity reaction (DHR) skin sites in guinea pigs. Listeria-elicited macrophages migrated toward MCF-a, -b, and -c. The chemotactic responses suggested responsive subpopulations to MCF. The electronic programmable individual cell sorter (EPICS) was used to separate macrophages with anti-la monoclonal antibodies. Ia-positive subpopulations responded to MCF-c, although they did not migrate toward MCF-a and -b. In contrast, Ia-negative subpopulations migrated toward MCF-a and -b, but not toward MCF-c. Furthermore, MCF-c attracted Ia-positive macrophages, whereas MCF-a and -b were Ia-negative in vitro; MCF did not induce Ia-negative macrophages to express surface Ia-antigens in vitro. MCF-c was able to produce massive Ia-positive macrophage accumulations when injected i.p., whereas MCF-a accumulated Ia-negative macrophages. The data suggest that MCF-a and -b, which mediate initial macrophage reactions, attract Ia-negative macrophages, and that MCF-c, which mediates predominant macrophage reactions, attract Ia-positive macrophages in the DHR.     

Effect of trace elements upon the biomass production of some candida species

Trace element studies were carried out on nine species of Candida. Out of twenty-three trace elements tested, Fe, Zn, Mn and Cu were found to be essential for the growth of all the yeast species whereas the rest of the elements exhibited variable essentiality. All the species of yeasts investigated required different concentrations of trace elements for their optimum growth. Concentrations higher than the optimum have been found to be inhibitory for the growth of all the yeasts studied here.     

Alveolar macrophage depletion is associated with increased surfactant pool sizes in adult rats.

Pulmonary surfactant is a lipid-protein material that is essential for normal lung function. Maintaining normal and consistent alveolar amounts of surfactant is in part dependent on clearance of surfactant by alveolar macrophages (AM). The present study utilized a rat model of AM depletion to determine the impact on surfactant pool sizes and function over time. Male Sprague-Dawley rats were anesthetized and intratracheally instilled with PBS-liposomes (PBS-L) or dichloromethylene diphosphonic acid (DMDP) containing liposomes (DMDP-L) and were killed at various time points up to 21 days for compliance measurements, AM cell counts, and surfactant analysis. AM numbers were significantly decreased 1, 2, and 3 days after instillation in DMDP-L vs. PBS-L, with 72% depletion at 3 days. AM numbers returned to normal levels by 5 days. In DMDP-L rats, there was a rapid increase in surfactant-phospholipid pools, showing a ninefold increase in the amount of surfactant in the lavage 3 days after liposome instillation. Surfactant accumulation progressed up to 7 days, with pools normalizing by 21 days. The increase in surfactant was due to increases in both subfractions of surfactant, the large aggregates (LA) and small aggregates. Surfactant protein A levels, relative to LA phospholipids, were not increased. There was a decreased extent of surfactant conversion in vitro for LA from DMDP-L rats compared with controls. It is concluded that the procedure of AM depletion significantly affects surfactant metabolism. The increased endogenous surfactant must be considered when utilizing the AM depletion model to study the role of these cells during lung insults.     

Alveolar macrophage activation after trauma-hemorrhage and sepsis is dependent on NF-kappaB and MAPK/ERK mechanisms

The acute respiratory distress syndrome (ARDS) is a major cause of morbidity after injury. We hypothesized that alveolar macrophage (AMPhi) chemokine and cytokine release after hemorrhage and sepsis is regulated by NF-kappaB and MAPK. Adult male rats underwent soft tissue trauma and hemorrhagic shock (~90 min) followed by crystalloid resuscitation. Sepsis was induced by cecal ligation and puncture (CLP) 20 h after resuscitation. AMPhi were harvested, and TNF-alpha, IL-6, and macrophage inflammatory protein (MIP)-2 release and serum IL-6 and TNF-alpha levels were measured at 5 h after HCLP. Lung tissues were analyzed for activation of NF-kappaB, myeloperoxidase activity, and wet/dry weight ratio. In control animals, AMPhi were stimulated with LPS with or without inhibitors of NF-kappaB and MAPK. Serum TNF-alpha and IL-6 levels and spontaneous AMPhi TNF-alpha and MIP-2 release were elevated (P < 0.05) after HCLP, concomitantly with the development of lung edema and leukocyte activation. Activation of NF-kappaB increased in lungs from the hemorrhage and CLP group compared with shams. Inhibition of NF-kappaB or the upstream MAPK significantly decreased LPS-stimulated AMPhi activation. Because enhanced release of inflammatory mediators by AMPhi may contribute to ARDS after severe trauma, inhibition of intracellular signaling pathways represents a target to attenuate organ injury under those conditions.     

Three new phylogenetic lineages are the closest relatives of the widespread species Albugo candida

White blister rust caused by the obligate biotroph Albugo candida (Albuginaceae; Oomycota) is one of the most notorious and common diseases of Brassicaceae. During the past 5 y, A. candida specimens collected from about 30 host genera were phylogenetically and morphologically investigated in several studies. These not only revealed that A. candida s.str. has a broad host range, encompassing a large number of host plants belonging to Brassicales, but also the presence of previously overlooked species of Albugo with hosts in this order. In this study, we examined specimens from Alyssum, Barbarea, and Rorippa, of which many species were commonly recorded as host plants of A. candida but could not be included in previous works due to the paucity of specimens available. It was revealed that Albugo specimens from Alyssum montanum, Barbarea vulgaris, and various Rorippa species, were placed in three phylogenetically distinct clades, but closer to A. candida s.str. than any previously reported species. Oospores were observed from Albugo specimens parasitic to Rorippa and could be distinguished morphologically from A. candida. Therefore, Albugo rorippae sp. nov. is described and illustrated here. In addition, a key of Albugo species described previously from Brassicales is given. The present study reveals that a large number of Albugo species remain still undiscovered, and that species close to A. candida exist. This could help elucidating the basis of the broad host range of A. candida as opposed to the narrow specialisation that is seemingly present in other species of Albugo on the Brassicaceae.     

Ascospore formation in yeasts related to Torulopsis candida

Ascospore formation was found in the type strains Torulopsis famata (Harrison) Lodder et Kreger van Rij, T. minor (Pollaci et Nannizi) Lodder and Candida flareri (Ciferri et Redaelli) Langeron et Guerra. The names of these species should be considered as a synonym of Debaryomyces hansenii (Zopf) Lodder et Kreger van Rij.     

Ecotoxicity of cobalt to the springtail Folsomia candida

Despite growing concern about the potential adverse effects of elevated cobalt concentrations in the environment, hardly any toxicity data are available for terrestrial invertebrates. Therefore, chronic toxicity of cobalt was assessed for the springtail Folsomia candida. The 28-day EC50 for the reproduction of F. candida was 1480 mg Co/kg dry wt in standard artificial soil (OECD) and 409 mg Co/kg dry wt in standard field soil (LUFA 2.2). The difference in toxicity can be explained by the higher pH and cation exchange capacity which decreased cobalt bioavailability in the OECD soil. When expressed as pore water concentrations, 28-day EC50s were similar: 159 mg Co/L in OECD and 174 mg Co/L in LUFA 2.2, which corresponded with calculated Co2+ activities of 0.953 and 1.20 mmol/L, respectively. Although the presented data can be considered as a step forward in the assessment of the potential risk of cobalt in the terrestrial environment, more toxicity data for different species are needed to evaluate the environmental risk of cobalt in soils.     

Fractionation of murine macrophage inhibitory factor into two distinct species

Murine migration inhibitory factor (MIF) produced by concanavalin A-stimulated lymph node cells from C57BL/6 mice was fractionated by Sephadex G-100 gel filtration, density gradient electrophoresis, and isoelectrofocusing in a sucrose density gradient and assayed on in vitro-cultivated bone marrow macrophages from C57BL/6 mice. Two major MIF species, pH3-MIF with an isoelectric point of 3.0–4.3 and pH5-MIF with an isoelectric point of 4.6 to 5.2, were obtained. The similarity of murine MIF to guinea pig and human MIF is discussed.