Biosorption efficiency of nickel by various endophytic bacterial strains for removal of nickel from electroplating industry effluents: an operational study.

Realising the hazardous effect of nickel on human health, microbes and plants are effectively used for bioremediation. The endophytic microorganisms have an important role in the phytoremediation of nickel using Vigna radiata. Therefore, in order to harness the potential of microbial strains, the present study was designed to examine the metal biosorption ability of endophytic bacterial strains isolated from plants growing in nickel-contaminated soil. A total of six endophytic nickel resistance bacteria were isolated from the plant Vigna radiata. The metal tolerant bacterial strains were identified following 16 S rRNA gene sequence analysis. Nickel biosorption estimation and plant growth-promoting (PGP) activities of isolated strains were performed and found high nickel biosorption efficiency of 91.3 ± 0.72% at 600 mg L-1 using Bacillus safensis an isolated endophytic strain from Vigna radiata. Furthermore, high indole acetic acid (IAA) and exopolysaccharide (EPS) production were obtained in all the strains as compared to without nickel-containing medium used as control. Moreover, the production of high EPS suggests improved biosorption ability of isolated endophytic strains. In addition, a kinetic study was also performed to evaluate different adsorptions isotherms and support the nickel biosorption ability of endophytic strains. The treatment of nickel electroplating industrial effluent was also demonstrated by isolated endophytic strains. Among six (6) strains, B. cereus showed maximum 57.2 ± 0.62% biosorption efficiency of nickel which resulted in the removal of 1003.50 ± 0.90 mg L-1 of nickel from the electroplating industry effluents containing initial 1791 ± 0.90 mg L-1 of nickel. All other strains were also capable of significant nickel biosorption from electroplating industry effluents as well. Thus, isolated endophytic nickel tolerant strains can be further used at large-scale biosorption of nickel from electroplating industry effluent.

IRAK1-dependent signaling mediates mortality in polymicrobial sepsis.

Interleukin-1 receptor-associated kinase (IRAK1) is a key regulatory protein in TLR/IL1R-mediated cell activation during inflammatory response. Studies indicated that pending on the nature of the used inflammatory model, downregulation of IRAK1 may be beneficial or detrimental. However, the role of IRAK1 in affecting outcome in polymicrobial sepsis is unknown. We tested this question using an IRAK1-deficient mouse strain and cecal ligation and puncture (CLP) procedure, which is a clinically relevant rodent septic model. Sepsis-induced mortality was markedly lower in IRAK1-deficient mice (35 %) compared to WT (85 %). Sepsis-induced increases in blood IL-6 and IL-10 levels were blunted at 6 h post-CLP in IRAK1 deficiency compared to WT, but cytokine levels were similar at 20 h post-CLP. Sepsis-induced blood granulocytosis and depletion of splenic B cells were also blunted in IRAK1-deficient mice as compared to WT. Analysis of TLR-mediated cytokine responses by IRAK1-deficient and WT macrophages ex vivo indicated a TLR4-dependent downregulation of IL-6 and IL1ß in IRAK1 deficiency, whereas TLR2-dependent responses were unaffected. TLR7/8-mediated IL-6, IL1ß, and IL-10 production was also blunted in IRAK1 macrophages as compared to WT. The study shows that IRAK1 deficiency impacts multiple TLR-dependent pathways and decreases early cytokine responses following polymicrobial sepsis. The delayed inflammatory response caused by the lack of IRAK1 expression is beneficial, as it manifests a marked increased chance of survival after polymicrobial sepsis.

Ecto-5'-nucleotidase (CD73) decreases mortality and organ injury in sepsis.

The extracellular concentrations of adenosine are increased during sepsis, and adenosine receptors regulate the host's response to sepsis. In this study, we investigated the role of the adenosine-generating ectoenzyme, ecto-5'-nucleotidase (CD73), in regulating immune and organ function during sepsis. Polymicrobial sepsis was induced by subjecting CD73 knockout (KO) and wild type (WT) mice to cecal ligation and puncture. CD73 KO mice showed increased mortality in comparison with WT mice, which was associated with increased bacterial counts and elevated inflammatory cytokine and chemokine concentrations in the blood and peritoneum. CD73 deficiency promoted lung injury, as indicated by increased myeloperoxidase activity and neutrophil infiltration, and elevated pulmonary cytokine levels. CD73 KO mice had increased apoptosis in the thymus, as evidenced by increased cleavage of caspase-3 and poly(ADP-ribose) polymerase and increased activation of NF-κB. Septic CD73 KO mice had higher blood urea nitrogen levels and increased cytokine levels in the kidney, indicating increased renal dysfunction. The increased kidney injury of CD73 KO mice was associated with augmented activation of p38 MAPK and decreased phosphorylation of Akt. Pharmacological inactivation of CD73 in WT mice using α, ß-methylene ADP augmented cytokine levels in the blood and peritoneal lavage fluid. These findings suggest that CD73-derived adenosine may be beneficial in sepsis.

Female X-chromosome mosaicism for NOX2 deficiency presents unique inflammatory phenotype and improves outcome in polymicrobial sepsis.

Cellular X-chromosome mosaicism, which is unique to females, may be advantageous during pathophysiological challenges compared with the single X-chromosome machinery of males, and it may contribute to gender dimorphism in the inflammatory response. We tested the hypothesis of whether cellular mosaicism for the X-linked gp91phox (NOX2) deficiency, the catalytic component of the superoxide anion-generating NADPH oxidase complex, is advantageous during polymicrobial sepsis. Deficient, wild-type (WT), and heterozygous/mosaic mice were compared following polymicrobial sepsis initiated by cecal ligation and puncture. Compared with WT littermates, sepsis-induced mortality was improved in deficient mice, as well as in mosaic animals carrying both deficient and WT phagocyte subpopulations. In contrast, blood bacterial counts were greatest in deficient mice. Consistent with poor survival, WT mice also showed the most severe organ damage following sepsis. In mosaic animals, the deficient neutrophil subpopulations displayed increased organ recruitment and elevated CD11b membrane expression compared with WT neutrophil subpopulations within the same animal. The dynamics of sepsis-induced blood and organ cytokine content and WBC composition changes, including lymphocyte subsets in blood and bone marrow, showed differences among WT, deficient, and mosaic subjects, indicating that mosaic mice are not simply the average of the deficient and WT responses. Upon oxidative burst, interchange of oxidants between WT and deficient neutrophil subpopulations occurred in mosaic mice. This study suggests that mice mosaic for gp91phox expression have multiple advantages compared with WT and deficient mice during the septic course.

Female X-chromosome mosaicism for gp91phox expression diversifies leukocyte responses during endotoxemia.

OBJECTIVE: To test the hypothesis, using an animal model, whether female X-chromosome mosaicism for inflammatory gene expression could contribute to the gender dimorphic response during the host response. X-chromosome-linked genetic polymorphisms present a unique biological condition because females display heterozygous cellular mosaicism, due to the fact that either the maternal or the paternal X chromosomes are inactivated in each individual cell in females. This is in contrast with the conditions in males who carry exclusively the maternal X chromosome. DESIGN: Prospective, randomized, laboratory investigation. SETTINGS: University research laboratory. SUBJECTS: Female mice deficient, heterozygous (mosaic) or WT for the X-linked gp91phox. INTERVENTIONS: We compared selected inflammatory markers among heterozygous (mosaics), WT and homozygous deficient animals in response to in vivo lipopolysaccharide (Escherichia coli, 20 mg/kg body weight). To test individual mosaic subpopulations of polymorphonuclear neutrophil responses, we also developed a flow cytometry assay that identifies the active parental X chromosomes in individual cells, using gp91phox expression as a marker. MEASUREMENTS AND MAIN RESULTS: Heterozygous mosaic mice presented white blood cell trafficking patterns similar to that observed in WT mice, despite the fact that the deficient subpopulation in mosaic animals displayed increased cell activation as reflected in elevated neutrophil CD11b expression and splenic infiltration. Mosaic animals also displayed splenic neutrophil infiltration, which was skewed toward the deficient subpopulation. Observations on splenic T-cell depletion and post lipopolysaccharide interleukin-10 responses indicated that the inflammatory response in mosaic animals does not simply display an average of the deficient and WT responses, but the mosaic subjects display a uniquely characteristic response. CONCLUSIONS: The study supports the notion that female X chromosome mosaicism for polymorphic gene expression represents a unique condition, which may contribute to the gender dimorphic character of the inflammatory response. Mosaicism for X-linked polymorphisms may have clinical significance and needs consideration in genetic association or gender-related clinical studies.

Endotoxemia down-regulates bone marrow lymphopoiesis but stimulates myelopoiesis: the effect of G6PD deficiency.

Bone marrow (BM) dysfunction is an important component of immunomodulation. This study investigated alterations in cell content, apoptotic responses, and cell proliferation in BM, blood, and spleen in endotoxemic mice (LPS from Escherichia coli). As the decreased antioxidant status associated with glucose-6-phosphate dehydrogenase (G6PD) deficiency has been shown to modulate the innate immune response, we also tested whether a G6PD mutation (80% decrease in cellular enzyme activity) alters BM responses during endotoxemia. LPS decreased BM myeloid (CD45(+)CD11b(+)) and B lymphoid (CD45(+)CD19(+)CD11b(-)) cell content compared with controls. In contrast, LPS increased CD11b(+) myeloid but decreased T and B cell counts in the circulation. Endotoxemia inhibited spontaneous, heat shock, and H(2)O(2)-induced apoptosis as well as proliferative activity in BM lymphoid cells. In contrast, BM myeloid cell apoptosis was not altered, and their proliferative activity was increased during endotoxemia. Following LPS, splenic myeloid cell content was increased, and T and B cell content was unchanged; furthermore, splenocytes showed increased apoptosis compared with controls. BM cell content, including lymphoid and myeloid cells, was greater in G6PD mutant than wild-type (WT) mice, and LPS decreased BM cell counts to a greater degree in mutant than WT mice. Endotoxemia caused widespread inhibition of BM cytokine and chemokine production; however, IL-6 production was increased compared with controls. LPS-induced IL-6 production was decreased in G6PD mutant animals compared with WT. This study indicates that endotoxin inversely affects BM myeloid and lymphoid cell production. LPS-induced down-regulation of B cell production contributes to the generalized lymphopenia and lymphocyte dysfunction observed following nonspecific immune challenges.

Flavopiridol blocks integrin-mediated survival in dormant breast cancer cells.

PURPOSE: Breast cancer micrometastases in the bone marrow are resistant to chemotherapy. They can remain dormant for years before some begin to proliferate. We seek to understand survival mechanisms and develop targeted approaches to eliminating these cells. EXPERIMENTAL DESIGN: In an in vitro model of dormancy, basic fibroblast growth factor 2 (FGF-2), abundant in the bone marrow, inhibits the growth of well-differentiated cells in the 2- to 10-cell stage and up-regulates integrin alpha(5)beta(1). Through this integrin, cells bind fibronectin, spread out, and acquire a survival advantage, partly through activation of the phosphatidylinositol 3-kinase/Akt pathway. We investigated the effects of Taxotere, flavopiridol, and mitogen-activated protein/extracellular signal-regulated kinase (ERK) kinase and p38 inhibitors on survival of dormant clones and that of flavopiridol on expression of integrins, adhesion strength, and phosphorylation of Akt, ERK 1/2, and p38. RESULTS: Dormant MCF-7 and T-47D cell clones were resistant to Taxotere concentrations 10-fold higher than needed to eliminate growing clones but were almost completely eradicated by 200 nmol/L flavopiridol. Flavopiridol caused a decrease in FGF-2-induced expression of integrins, including alpha(5) and beta(1), and decreased FGF-2-induced specific adhesion to fibronectin. It diminished Akt phosphorylation, but reexpression of active Akt was not sufficient to reverse dormant clone inhibition. Flavopiridol did not affect phosphorylation of ERK 1/2 and p38 but diminished total protein levels. Chemical inhibition of these pathways partially abrogated dormant clone survival. CONCLUSIONS: Flavopiridol has pleiotropic effects on key targets involved with survival of dormant breast cancer cells and may represent a useful approach to eliminating cells dependent on multiple signal pathways for survival.

Phenotypic & genotypic variants of Pseudomonas aeruginosa isolated from children with cystic fibrosis in India.

BACKGROUND & OBJECTIVES: Pseudomonas aeruginosa is the leading cause of morbidity and mortality in patients with cystic fibrosis (CF). With increase in the chronicity of the disease, there is a diversification of the organism into different colony morphological types. The antimicrobial susceptibility of the organism varies with its colony morphology. The present work was carried out to study the different morphotypes of P. aeruginosa isolated from patients of cystic fibrosis. METHODS: We studied 38 children with CF attending the Paediatric Chest Clinic at the All India Institute of Medical Sciences, New Delhi, India during October 2000-January 2001 who were regularly followed up at the clinic. Patients were divided into 2 groups, Group 1 included all patients chronically infected with P. aeruginosa and Group 2 included patients who were infrequently colonized with this organism. Different colony morphological types of P. aeruginosa on culture media were identified. They were characterized by phenotypic methods using antibiograms and genotypic methods using enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction and PCR-ribotyping. RESULTS: Fourteen of the 38 patients were colonized at least once with P. aeruginosa. Eight patients belonged to Group 1 and 42 isolates were obtained from these patients. Group 2 had 6 patients and 9 isolates were obtained from them. All patients in Group 1 harboured different colony morphotypes (Types 1-6) while all 6 patients in Group 2 showed a single type of colony morphology (Type 1). The isolates from Group 1 patients showed higher antimicrobial resistance as compared to Group 2 patients. Molecular typing of the isolates revealed 10 ERIC-PCR patterns and 2 PCR-ribotyping patterns among Group 1 and 2 ERIC-PCR and 1 PCR-ribotyping pattern among patients of Group 2. INTERPRETATION & CONCLUSION: The frequency of different morphotypes of P. aeruginosa and antibiotic resistance was higher among Group 1 patients. On molecular typing, more than one genotype was isolated from Group 1 patients while only one genotype was isolated from patients in Group 2. We conclude that at a given time, chronically infected patients can be colonized by phenotypically and genotypically distinct strains of P. aeruginosa which has an implication in the management of these patients.