Urban wastelands: On the frontline between air pollution sources and residential areas.

In urban areas, particulate matter (PM) represents an increasing threat to human health. The ability of plants in parks and along roads in cities to accumulate PM has already been demonstrated, but nothing is known about the effect of wasteland vegetation on air quality, despite a significant proportion of greenery in polluted areas being on wastelands. The aim of this study was to document the accumulation of PM and trace elements (TE) by wasteland species (Robinia pseudoacacia L., Populus × canescens (Aiton) Sm., Acer negundo L., Solidago gigantea (Aiton) and Poaceae) growing on Central European urban wastelands with differing levels of air pollution. On average, the largest amounts of PM accumulated on the foliage of R. pseudoacacia and S. gigantea, and the smallest amounts accumulated on P. × canescens leaves. However, accumulation of PM depended more on the distance from the emission source than on species selection, and was higher on the polluted wasteland where the plants' gas exchange was the lowest. The results also suggest that in order to effectively accumulate PM from the air, it is critical to have the correct configuration of plants, with the wasteland vegetation having a layered structure and layers differing in PM retention, as shown in this study using the examples of R. pseudoacacia (a tall tree with low PM retention) and S. gigantea (below-tree vegetation with high PM retention). P. × canescens accumulated the highest concentrations of Cd and Zn, S. gigantea accumulated the highest concentration of Cu, and Poaceae accumulated the highest concentrations of Cr and Ni. These findings have implications for urban vegetation management in areas where there is no organised greenery, and offer proof that vegetation in wasteland areas should be maintained since it is an excellent tool for reducing concentrations of PM at its place of origin.

Physical fitness and nutritional status of polish ground force unit recruits.

The purpose of the work was to conduct an examination of the physical fitness and nutritional status of recruits (221 men beginning military service in the infantry unit). Soldiers' physical efficiency was estimated using 4 tests: standing long jump, pull-ups on bar, 30-second sit-ups and 1000-metre run. The nutritional status assessment was done based on anthropometric measurements including measurements of body height, body mass and selected skin fold thickness. The study group of soldiers were the best at sit-ups (46.33 points). They got over 40 points for the 1000-metre run (43.68 points) and for pull-ups on bar (41.69 points). They obtained the lowest scores for standing long jumps (30.77 points). About 14% of recruits were overweight and 4.1% underweight. Recruits enrolling in the infantry unit present a low physical fitness level. Overweight and obesity occurrence, and particularly underweight, in recruits testify to improper nutrition before beginning military service.

The influence of folic acid, vitamins B(2) and B(6) supplementation on feed intake, body and organs weight, and liver fatty acids composition of rats subjected to 3 months moderate protein deprivation.

This study was conducted to determine the effect of a 3-month dietary protein restriction - protein provided 9% of energy (20% in control group). In this dietary restriction folic acid, vitamins B(2) and B(6) were delivered in amount three times above the standard level. It was observed that animals fed a protein restricted (PR) diet weighed about 5% less than animals consuming adequate diet, but the difference was not statistically significant. Enrichment of PR diet with vitamin B or folic acid caused tendency to further suppression of weight gain, and in case of vitamin B(6) these differences were statistically significant. However, such body weight (BW) suppression was not observed when all studied vitamins were used together. Significant reductions in relative liver weight (vitamin B(2) addition), the heart (folic acid) and the lungs (vitamin B(6)) were observed. The PR diet, when all vitamins were added together, caused a decrease in weights of the lungs, heart and liver scaled to BW of rats, simultaneously with a significant increase in testis weight. Feed intake and feed conversion ratio were higher in animals given PR diet without a significant influence of vitamin supplementation (except vitamin B(6) causing further increase in feed conversion ratio). Hepatic fatty acids composition of rats was not affected by protein restriction, as well as by single vitamin supplementation. However, dietary supplementation of all examined vitamins together caused a decrease in monounsaturated fatty acids followed by an increase in polyunsaturated fatty acids participation in total fatty acids pool. It seems that enrichment of PR diet with a mixture of folic acid, vitamins B(2) and B(6) resulted in a partial reverse of growth suppression and reduction in testis size in rats.

Influence of folic acid, vitamin B2 and B6 supplementation on feed intake, body and organs weight, and liver fatty acids composition in rats subjected to severe protein deprivation.

Growing rats fed for 3 months a low-protein (LP) diet (4.5% of energy from protein), possessed about 29% lower body weight than animals consuming adequate-protein diet (20% energy from protein). The LP diet feeding caused an increase in daily feed intake followed by a decrease in feed conversion efficiency. The enrichment of LP diet with folic acid, vitamin B2 and B6 (3 times above the level applied in the control diet) did not have any impact on rats BW and supplementation with these vitamins minimize the effect of LP diet on feed intake. The use of examined vitamins had a tendency to diminish an increase in feed conversion ratio caused by the LP nutrition. This effect was significant when all vitamins were added together. Rats fed the LP diet had higher relative weights of lungs, heart, liver and testis. Vitamins enriching the LP diet were observed to decrease a relative weight of lungs (folic acid, vitamin B6 and vitamin mixture), and liver (vitamin B6 and vitamin mixture). A tendency of increasing relative testis weight was also revealed in rats given the LP diet enriched with vitamins. The lower content of hepatic polyunsaturated fatty acids (FA) and a tendency for monounsaturated FA content to be higher were found in rats fed the LP diet. The LP diet enrichment with folic acid caused that these changes were more pronounced and statistically significant. Enrichment of LP diet with vitamins tested may cause a partial reverse of changes observed in the hepatic FA composition.

Biocompatibility parameters of different dialysis membranes assessed during systemic inflammation.

BACKGROUND: We explored whether biocompatible dialyzer membranes modulate the inflammatory response during blood contact in patients with systemic inflammation. METHODS: 15 patients with end-stage renal disease and systemic inflammation (mean serum C-reactive protein 86 +/- 4 mg/l) were randomly treated with Cuprophan (CU), polyamide (PA) and vitamin-E coated (VEC) membrane-based dialyzers. RESULTS: Changes in blood pressure, capillary blood oxygen saturation and differential blood counts during the hemodialysis session were not significantly different between the three dialyzers. Baseline blood levels of activated circulating complement (C3a) were more than 100 times above normal, and unlike expected they decreased during hemodialysis treatments (CU: from 7,389 +/- 783 to 5,423 +/- 761 ng/ml; PA: from 7,379 +/- 980 to 5,690 +/- 714 ng/ml; VEC: from 7.377 +/- 714 to 5,360 +/- 1,005 ng/ml; all n.s.). No significant differences between treatments were found with respect to changes in blood concentrations of TNF-alpha, interleukin-6 and interleukin-1 receptor antagonist as well as ICAM-1 (CU: from 451 +/- 41 to 477 +/- 41 ng/ml; PA: from 437 +/- 42 to 449 +/- 40 ng/ml; VEC: from 461 +/- 43 to 460 +/- 47 ng/ml). Furthermore, generation of reactive oxygen species by mononuclear blood cells was comparable during hemodialysis with the CU, PA and VEC dialyzer. CONCLUSION: The choice of dialyzer membrane material does not affect most aspects of biocompatibility when patients have significant systemic inflammation. This confounding variable should be taken into account in studies exploring the effects of biocompatible dialyzer membranes.

The reprogrammed host: Chlamydia trachomatis-induced up-regulation of glycoprotein 130 cytokines, transcription factors, and antiapoptotic genes.

OBJECTIVE: Infection with Chlamydia trachomatis is a known cause of sexually transmitted diseases, eye infections (including trachoma), and reactive arthritis (ReA). Because the mechanisms of Chlamydia-induced changes leading to ReA are poorly defined, this study sought to identify the target genes involved at the molecular level. METHODS: Chlamydia-induced changes in host cells were investigated by combining a screening technique, which utilized complementary DNA arrays on C trachomatis-infected and mock-infected epithelial HeLa cells, with real-time reverse transcription-polymerase chain reaction or enzyme-linked immunosorbent assay of gene products. Some responses were additionally demonstrated on human primary chondrocytes and a human synovial fibroblast cell line, both of which served as model cells for ReA. RESULTS: Eighteen genes (of 1,176) were found to be up-regulated after 24 hours of infection with this obligate intracellular bacterium, among them the glycoprotein 130 family members IL-11 and LIF, the chemokine gene MIP2-alpha, the transcription factor genes EGR1, ETR101, FRA1, and c-jun, the apoptosis-related genes IEX-1L and MCL-1, adhesion molecule genes such as ICAM1, and various other functionally important genes. In the context of this rheumatic disease, the cytokines and transcription factors seem to be especially involved, since various connections to chondrocytes, synoviocytes, bone remodeling, joint pathology, and other rheumatic diseases have been demonstrated. CONCLUSION: Infection with C trachomatis seems to reprogram the host cells (independent of activation by lipopolysaccharide or other ultraviolet-resistant bacterial components) at various key positions that act as intra- or intercellular switches, suggesting that these changes and similar Chlamydia-induced functional alterations constitute an important basis of the pathogenic inflammatory potential of these cells in ReA. Our results suggest that this approach is generally useful for the broad analysis of host-pathogen interactions involving obligate intracellular bacteria, and for the identification of target genes for therapeutic intervention in this rheumatic disease.

Identification of a selective nonpeptide antagonist of the anaphylatoxin C3a receptor that demonstrates antiinflammatory activity in animal models.

The anaphylatoxin C3a is a potent chemotactic peptide and inflammatory mediator released during complement activation which binds to and activates a G-protein-coupled receptor. Molecular cloning of the C3aR has facilitated studies to identify nonpeptide antagonists of the C3aR. A chemical lead that selectively inhibited the C3aR in a high throughput screen was identified and chemically optimized. The resulting antagonist, N(2)-[(2,2-diphenylethoxy)acetyl]-L-arginine (SB 290157), functioned as a competitive antagonist of (125)I-C3a radioligand binding to rat basophilic leukemia (RBL)-2H3 cells expressing the human C3aR (RBL-C3aR), with an IC(50) of 200 nM. SB 290157 was a functional antagonist, blocking C3a-induced C3aR internalization in a concentration-dependent manner and C3a-induced Ca(2+) mobilization in RBL-C3aR cells and human neutrophils with IC(50)s of 27.7 and 28 nM, respectively. SB 290157 was selective for the C3aR in that it did not antagonize the C5aR or six other chemotactic G protein-coupled receptors. Functional antagonism was not solely limited to the human C3aR; SB 290157 also inhibited C3a-induced Ca(2+) mobilization of RBL-2H3 cells expressing the mouse and guinea pig C3aRS: It potently inhibited C3a-mediated ATP release from guinea pig platelets and inhibited C3a-induced potentiation of the contractile response to field stimulation of perfused rat caudal artery. Furthermore, in animal models, SB 290157, inhibited neutrophil recruitment in a guinea pig LPS-induced airway neutrophilia model and decreased paw edema in a rat adjuvant-induced arthritis model. This selective antagonist may be useful to define the physiological and pathophysiological roles of the C3aR.

[Complications of three methods of stapedectomy]. / Powiklania trzech metod stapedektomii.

In a group of 270 partial stapedectomies performed by one surgeon using teflon-piston prosthesis it was assessed whether incidence of complications depended on sequence of surgical steps. In group I of 50 ears classical stapedectomy was performed: 1. removal of stapes superstructure, 2. making a hole in the footplate, 3. placing the prosthesis on the incus. In group II of 167 ears the hole in the footplate was made before removal of stapes arch. In group III of 53 ears: 1. the hole in the footplate, 2. placing the prosthesis on the incus, 3. removal of stapes arch, was performed. Total deafness had one patient of the whole group. No patient had perilymphatic fistula and facial nerve palsy. Floating footplate developed in 4 patients of group I, in one patient of group II and in none patient of group III. Subluxation of incus developed in 3 patients of group I, in 12 patients of group II and in none patient of group III. Comparison of three stapedectomy methods showed that making the hole in the footplate before removal of stapes arch prevents floating footplate and placing the prosthesis on the incus before removal of stapes arch prevents subluxation of the incus.

[Free forearm skin flap with microvascular anastomosis for oral cavity reconstruction]. / Wolny plat skóry przedramienia z mikrozespoleniem naczyn do rekonstrukcji jamy ustnej.

Free forearm skin-fascia flap was used in 8 patients for intraoral reconstruction. Tissue defect was located in anterior part of the oral cavity in three patients and in lateral part in 5 patients. In the first three patients microvascular anastomosis was performed first and the flap was sutured in the oral cavity later on. In one of these patients the whole flap and in two patients about half of the flap necrotised. In the next 5 patients oral defect was closed first and then microanastomosis was done with good result in all the patients. We advise the last type of reconstruction. Free forearm skin-fascia flap provides good anatomical and functional result of reconstruction in the oral cavity.

Cutting edge: guinea pigs with a natural C3a-receptor defect exhibit decreased bronchoconstriction in allergic airway disease: evidence for an involvement of the C3a anaphylatoxin in the pathogenesis of asthma.

Asthma is a major cause of morbidity worldwide with prevalence and severity still increasing at an alarming pace. Hallmarks of this disease include early-phase bronchoconstriction with subsequent eosinophil infiltration, symptoms that may be mimicked in vivo by the complement-derived C3a anaphylatoxin, following its interaction with the single-copy C3aR. We analyzed the pathophysiological role of the C3a anaphylatoxin in a model of experimental OVA-induced allergic asthma, using an inbred guinea pig strain phenotypically unresponsive to C3a. Molecular analysis of this defect revealed a point mutation within the coding region of the C3aR that creates a stop codon, thereby effectively inactivating gene function. When challenged by OVA inhalation, sensitized animals of this strain exhibited a bronchoconstriction decreased by approximately 30% in comparison to the corresponding wild-type strain. These data suggest an important role of C3a in the pathogenesis of asthma and define a novel target for drug intervention strategies.

[Operations of parotid malignant tumors]. / Operacje nowotworów zlosliwych slinianek przyusznych.

In a group of 64 patients with malignant tumours of the parotid gland the most common type was adenoid cystic carcinoma (20%) and malignant pleomorphic adenoma (14%). A group of 33 patients with follow-up from 4 months to 18 years was analyzed. The overall 5-year survival rate was 45%.

Discrimination of sepsis and systemic inflammatory response syndrome by determination of circulating plasma concentrations of procalcitonin, protein complement 3a, and interleukin-6.

OBJECTIVE: To evaluate whether plasma concentrations of procalcitonin (PCT), interleukin-6 (IL-6), protein complement 3a (C3a), leukocyte elastase (elastase), and the C-reactive protein (CRP) determined directly after the clinical onset of sepsis or systemic inflammatory response syndrome (SIRS) discriminate between patients suffering from sepsis or SIRS and predict the outcome of these patients. DESIGN: Prospective study. SETTING: Medical intensive care unit at a university hospital. PATIENTS: Twenty-two patients with sepsis and 11 patients with SIRS. MEASUREMENTS AND MAIN RESULTS: The plasma concentrations of PCT, C3a, and IL-6 obtained < or =8 hrs after clinical onset of sepsis or SIRS but not those of elastase or CRP were significantly higher in septic patients (PCT: median, 16.8 ng/mL, range, 0.9-351.2 ng/mL, p = .003; C3a: median, 807 ng/mL, range, 422-4788 ng/mL, p < .001; IL-6: median, 382 pg/mL, range, 5-1004 pg/mL, p = .009, all Mann-Whitney rank sum test) compared with patients suffering from SIRS (PCT: median, 3.0 ng/mL, range, 0.7-29.5 ng/mL; C3a: median, 409 ng/mL, range, 279566 ng/mL; IL-6: median, 98 pg/mL, range, 23-586 pg/mL). The power of PCT, C3a, and IL-6 to discriminate between septic and SIRS patients was determined in a receiver operating characteristic analysis. C3a was the best variable to differentiate between both populations with a maximal sensitivity of 86% and a specificity of 80%. An even better discrimination (i.e., a maximal sensitivity of 91% and a specificity of 80%) was achieved when PCT and C3a were combined in a "sepsis score." C3a concentrations also helped to predict the outcome of patients. Based on the sepsis score, a logistic regression model was developed that allows a convenient and reliable determination of the probability of an individual patient to suffer from sepsis or SIRS. CONCLUSIONS: Our data show that the determination of PCT, IL-6, and C3a is more reliable to differentiate between septic and SIRS patients than the variables CRP and elastase, routinely used at the intensive care unit. The determination of PCT and C3a plasma concentrations appears to be helpful for an early assessment of septic and SIRS patients in intensive care.

[Free forearm skin flaps with microvascular anastomosis for pharyngeal reconstruction]. / Wolny plat skóry przedramienia z mikrozespoleniem naczyn do rekonstrukcji gardla.

Free forearm skin flap with microvascular anastomosis was used for tissue defect reconstruction in 4 patients after resection of the oropharynx, base of the tongue and ramus of the mandible. The vascular pedicle of the flap contained the radial artery and the cephalic vein. For microvascular anastomosis the facial artery was used in all the patients, the facial vein in 2 patients, the internal jugular vein in one patient and the external jugular vein in the other one. Good healing of the graft was obtained in 3 patients. In one patient partial necrosis developed. No patient had fistula.

[Pharyngeal closure after pharyngolaryngectomy]. / Zamykanie gardla po faryngolaryngektomii.

Pharyngolaryngectomy was performed in 53 patients. In 36 patients pharynx defect was less than 50% of pharynx circumference and it was closed without reconstruction. Larger pharynx defects were closed using platysma myocutaneous flap (13 cases), pectoralis mayor flap (3 cases) and free forearm flap with microvascular anastomosis (1 case). Healing results are presented in each group of patients.

Guinea pig C3 specific rabbit single chain Fv antibodies from bone marrow, spleen and blood derived phage libraries.

We constructed combinatorial immunoglobulin libraries from the whole rabbit antibody repertoire of bone marrow, spleen and peripheral blood of a rabbit immunized with guinea pig complement protein C3. By means of the phage display technology we selected guinea pig C3 specific single chain Fv (scFv) antibodies from each of the libraries. None of the scFv antibodies cross reacted with guinea pig C3a, human C3 or rat C3. The frequency of bone marrow derived C3 positive clones was much higher as compared to blood or spleen derived clones. Additionally bone marrow and spleen derived clones show higher diversity than clones, obtained from blood, as determined by fingerprint analysis with the restriction enzyme AluI. Dissociation rate constants for all scFvs were similar, indicating that the source of the scFvs had no influence on affinities. The antibody fragments were used to analyze complement activation during xenotransplantation. Several blood or bone marrow derived scFvs bound to C3 located on rat liver endothelium after hyperacute rejection of a heterotopically transplanted rat liver into guinea pig. These data demonstrate that monoclonal rabbit scFvs can be easily generated from recombinant phage display libraries, constructed from spleen, blood or bone marrow. The selected guinea pig C3 specific scFvs appear to be useful to detect complement activation during xenotransplantation in guinea pigs.

Minor role of the C3a receptor in systemic anaphylaxis in the guinea pig.

Previously, Regal et al. [Regal, J.F., Fraser, D.G., Toth, C.A., 1993. Role of the complement system in antigen-induced bronchoconstriction and changes in blood pressure in the guinea pig. J. Pharmacol. Exp. Ther. 267, 979-988] demonstrated that preventing complement system activation resulted in inhibition of anaphylaxis in the guinea pig, and that the C-terminal 21 amino acids of guinea pig C3a (C3a-peptide) mimic the symptoms of anaphylactic shock in the guinea pig [Regal, J.F., 1997. Role of the complement system in pulmonary disorders. Immunopharmacology 38, 17-25]. To determine if C3a is an essential mediator of systemic anaphylaxis, the anaphylactic response to ovalbumin (OA) was assessed in guinea pigs genetically deficient in the C3a receptor (C3aR-) compared to their control strain of animals which were C3a receptor positive (C3aR+). In addition, the response to another control strain of animals, Hartley guinea pigs, was determined. Sensitized guinea pigs were anesthetized, and bronchoconstriction and changes in blood pressure were monitored in response to intravenous (i.v.) injection of either C3a-peptide, recombinant human C5a (rHuC5a) or OA. Both Hartley guinea pigs and C3aR+ animals responded similarly to C3a-peptide and rHuC5a. C3aR- animals, however, were unresponsive to C3a-peptide and responded normally to rHuC5a, confirming their functional deficiency of the C3a receptor. In response to OA, C3aR+ animals and Hartley guinea pigs responded with a severe bronchoconstriction, an initial transient hypotension, followed by an increase in blood pressure and a delayed prolonged hypotensive response. In contrast, in C3aR- animals, the increased blood pressure response to OA was significantly prolonged, the delayed hypotensive response was blunted, and the bronchoconstriction was delayed compared to the C3aR+ animals. The difference in the anaphylactic response could not be explained by differing amounts of OA-specific IgG1 antibody or C3a generated during the anaphylactic response. Thus, these data suggest that C3a plays a minor role in the hypotension of systemic anaphylaxis and investigation of a role for other products of complement system activation, either alone or in combination with C3a, is clearly warranted.

Selection of a C5a receptor antagonist from phage libraries attenuating the inflammatory response in immune complex disease and ischemia/reperfusion injury.

A C5a-receptor antagonist was selected from human C5a phage display libraries in which the C terminus of des-Arg74-hC5a was mutated. The selected molecule is a competitive C5a receptor antagonist in vitro and in vivo. Signal transduction is interrupted at the level of G-protein activation. In addition, the antagonist does not cause any C5a receptor phosphorylation. Proinflammatory properties such as chemotaxis or lysosomal enzyme release of differentiated U937 cells, as well as C5a-induced changes in intracellular Ca2+ concentration of murine peritoneal macrophages, are inhibited. The in vivo efficacy was evaluated in three different animal models of immune complex diseases in mice, i.e., the reverse passive Arthus reaction in the peritoneum, skin, and lung. The i.v. application of the C5a receptor antagonist abrogated polymorphonuclear neutrophil accumulation in peritoneum and markedly attenuated polymorphonuclear neutrophil migration into the skin and the lung. In a model of intestinal ischemia/reperfusion injury, i.v. administration of the C5a receptor antagonist decreased local and remote tissue injury: bowel wall edema and hemorrhage as well as pulmonary microvascular dysfunction. These data give evidence that C5a is an important mediator triggering the inflammatory sequelae seen in immune complex diseases and ischemia/reperfusion injury. The selected C5a receptor antagonist may prove useful to attenuate the inflammatory response in these disorders.

Analysis of the C5a anaphylatoxin core domain using a C5a phage library selected on differentiated U937 cells.

The human anaphylatoxin C5a is a 74-amino acid comprising polypeptide with a plethora of biological functions. Site directed mutagenesis studies suggest that several residues within the core and the C-terminus mediate the interaction with the C5a receptor. However, the contribution of particular core residues to receptor binding remained to be clarified. By means of the phage display technique, the loop between positions 35-40 was randomly mutated and the resulting C5a[35-40] fusion phage library affinity selected on C5a receptor expressing U937 cells. After five rounds of affinity enrichment, residues Arg37 and Arg40 were preferably selected. Enrichment was as high as 100% for Arg37 and 79% for Arg40. No significant enrichment of consensus residues could be obtained for positions 35, 36, 38 and 39. The core mutant C5a[A35E36R37A38S39R40], in which only Arg37/40 and Ala38 are of the native C5a sequence, was as potent as native C5a in both receptor binding and enzyme release examined on U937 cells. In contrast, replacement of Arg40 as in the mutant C5a[Q35E36R37I38L39N40] resulted in a 10-fold decrease in both binding and functional activities. Thus, selected out of a multiplicity of possibilities by the natural binding partner, Arg37 as well as Arg40 appear to be anchor residues in binding to the C5a receptor.