Biodegradation of nitroglycerin in porous media and potential for bioaugmentation with Arthrobacter sp. strain JBH1.

Nitroglycerin (NG) is a toxic explosive found as a contaminant of soil and groundwater. Several microbial strains are capable of partially reducing the NG molecule to dinitro or mononitroesters. Recently, a strain capable of growing on NG as the sole source of carbon and nitrogen (Arthrobacter sp. strain JBH1) was isolated from contaminated soil. Despite the widespread presence of microbial strains capable of transforming NG in contaminated soils and sediments, the extent of NG biodegradation at contaminated sites is still unknown. In this study column experiments were conducted to investigate the extent of microbial degradation of NG in saturated porous media, specifically after bioaugmentation with JBH1. Initial experiments using sterile, low sorptivity sand, showed mineralization of NG after bioaugmentation with JBH1 in the absence of sources of carbon and nitrogen other than NG. Results could be modeled using a first order degradation rate of 0.14d(-1). Further experiments conducted using contaminated soil with high organic carbon content (highly sorptive) resulted in column effluents that did not contain NG although high dinitroester concentrations were observed. Bioaugmentation with JBH1 in sediments containing strains capable of partial transformation of NG resulted in complete mineralization of NG and faster degradation rates.

In elderly women moderate hypercholesterolemia is associated to endothelial and microcirculatory impairments.

How cholesterol influences the microcirculation on aging subjects is not well known. This study evaluated moderate hypercholesterolemia effects in, treated or not, lean elderly women on brachial artery reactivity and microcirculatory function using venous occlusion plethysmography (VOP) and nailfold videocapillaroscopy (NVC). Patients (mean age 73 years) were divided into healthy elderly (HE, n=15), treated dyslipidemia with statins during at least 6 months (TDL, n=9) and dyslipidemia (DL, n=9, cholesterol, 257±11 and LDL-cholesterol, 157±24 mg/dl). Young, mean age 23 years, women (YC, n=24), served as controls. Laboratory and anthropometrical analysis, VOP peak forearm blood flow (FBF) during the reactive hyperemia response/baseline FBF (%HYPER) and peak FBF after 0.4 mg sublingual nitroglycerin/baseline FBF (%NITRO) were assessed. NVC capillary density and diameters, maximum red blood cell velocity (RBCV(max)) during reactive hyperemia/baseline RBCV and time to reach RBCV(max) were evaluated. Correlations between %HYPER, %NITRO and plasma cholesterol fractions were performed. Total and LDL-cholesterol were increased only in DL group. Capillary diameters were larger in elderly groups than YC. RBCV(max)/baseline RBCV was reduced in the DL group compared to HE, TDL and YC. %HYPER was lower in DL and normalized in TDL group. YC %HYPER was double of HE. %NITRO decreased from (HE=YC) to TDL and DL groups. There was a significant inverse correlation between LDL-cholesterol, non-HDL-cholesterol and %HYPER/% Nitro. In conclusion, moderate hypercholesterolemia reversibly impaired the vasodilatatory response in the microcirculation but the endothelial-independent vasodilator response to nitroglycerine remained irreversibly lower in healthy aged women.

Key enzymes enabling the growth of Arthrobacter sp. strain JBH1 with nitroglycerin as the sole source of carbon and nitrogen.

Flavoprotein reductases that catalyze the transformation of nitroglycerin (NG) to dinitro- or mononitroglycerols enable bacteria containing such enzymes to use NG as the nitrogen source. The inability to use the resulting mononitroglycerols limits most strains to incomplete denitration of NG. Recently, Arthrobacter strain JBH1 was isolated for the ability to grow on NG as the sole source of carbon and nitrogen, but the enzymes and mechanisms involved were not established. Here, the enzymes that enable the Arthrobacter strain to incorporate NG into a productive pathway were identified. Enzyme assays indicated that the transformation of nitroglycerin to mononitroglycerol is NADPH dependent and that the subsequent transformation of mononitroglycerol is ATP dependent. Cloning and heterologous expression revealed that a flavoprotein catalyzes selective denitration of NG to 1-mononitroglycerol (1-MNG) and that 1-MNG is transformed to 1-nitro-3-phosphoglycerol by a glycerol kinase homolog. Phosphorylation of the nitroester intermediate enables the subsequent denitration of 1-MNG in a productive pathway that supports the growth of the isolate and mineralization of NG.