Physiological responses of Bituminaria bituminosa to heavy metals

Two hydroponic experiments were performed to study the physiological responses to heavy metals (HMs) of two populations of Bituminaria bituminosa (L.) C.H. Stirton (Fabaceae): one (“C2”) from a site contaminated by HMs and one from a non-contaminated site (“LA”). In the first, we studied the effects of elevated concentrations of Zn (12 and 61 μM). Population C2 was more tolerant in terms of root and shoot growth at 61 μM Zn, relative to control plants (1 μM Zn). The similar tissue Zn levels of the two populations suggest that C2 is more tolerant of high tissue Zn. Of the parameters measured that could be related to Zn phytotoxicity (micro and macronutrients, root hydraulic activity, water-extractable Zn and organic acids), none could explain totally the superior tolerance of C2. In the second assay, the effects of Cd (4.4 μM), Cu (7.8 μM) and Zn (76 μM) on plant accumulation of the furanocoumarins (FCs) psoralen and angelicin, which function as feeding deterrents and photo-activated toxins, were assessed. For population C2, all three HMs increased the root FC concentrations, while Cd also raised shoot levels. For LA, Cu raised the root concentrations of both FCs. There was a relationship between plant stress, manifested as proline accumulation and disruption of plant water relations, and increased FC accumulation. Higher tissue levels of FCs likely provide greater protection against bacterial or fungal infection and herbivores.     

New avenues for phage-display library to produce a Cryptococcus-specific anti-idiotypic antibody of HM-1 killer toxin

Existing antifungal drugs are notable for their inability to act rapidly, as well as their toxicity and limited spectrum. The identification of fungal-specific genes and virulence factors would provide targets for new and influential drugs. The display of repertories of antibody fragments on the surface of filamentous phage offers a new way to produce immunoreagents as defined specificities. Here we report the selection of Cryptococcus-specific targets by using phage-display panning from a cDNA library, where bactericidal antibodies have been developed against conserved surface-exposed antigens. A single-chain variable fragment (scFv) phage library was constructed from splenocyte of an immunized mouse by idiotypic vaccination with HM-1 killer toxin (HM-1) neutralizing monoclonal antibody (nmAb-KT) that was used for selection against Cryptococcus neoformans membrane fraction (CnMF). Key elements were the selection against antigen (nmAb-KT and CnMF) and the release of bound phages using competitive panning elution with CnMF at neutral pH condition. Isolated scFvs react specifically with C. neoformans and some other pathogenic and non-pathogenic fungal strain's cell wall receptors by exerting strong antifungal activity in vitro. A high affinity clone, designated M1 was selected for detailed characterization and tested anti-cryptococcal activity with IC(50) values at 5.33 × 10(-7) to 5.56 × 10(-7) M against C. neoformans. The method described here is a new technique for the isolation of cell membrane specific immunoreactive phages in the form of scFv using CnMF that contained cell membrane associated proteins.     

Origin of the First Cells on Earth: A Possible Scenario

A fundamental challenge in science is to understand the assembly of the first macromolecules necessary for cellular life and the order in which this occurred. The assembly events that led to the first minimal cell capable of growth and division are a highly debatable subject. Possibly, the formation of a primitive membrane or microsphere in a hydrophobic medium provided a suitable structure where subsequent biochemical self-replication and eventually enzyme catalysis, integrated biochemical pathways, and assembly of nucleic acids occurred. In this article, I examine a possible sequence of assembly for the first primitive cell(s) on the Earth.     

Integrating process engineering and microbiology tools to advance activated sludge wastewater treatment research and development

Wastewater treatment is a huge industryworldwide. Despite the massive capital andoperating costs, only a relatively small amountof R&D investment is made. This might havebeen related to the limited demands in terms ofeffluent quality in the past, but today'senvironmental awareness requires much strictereffluent standards to be achieved. This inturn should give sufficient incentives,together with the possible large cost savings,to increase the R&D activities in this field. There are certainly significant knowledge gapsto be filled and substantial benefits could begained from this.A range of knowledge gaps are identified inthis paper, extending from the role ofintermediates in nutrient removal overparameter estimation in modelling andsimulation to understanding the microbialmetabolic pathways at a genetic and enzymaticlevel. These gaps are opportunities andchallenges for all researchers andprofessionals in this field. Addressing themwill help substantially in the continuingdevelopment of wastewater treatmenttechnologies.The complexity of biological wastewatertreatment processes requires a broad range oftools and expertise to address the knowledgegaps. Novel process analysis tools arecritically important to investigate biologicaltreatment processes in future. They will comefrom different expertise areas and will need tobe used in close integration to gain maximalbenefits from the efforts. These tools willlikely include respirometry, novel chemicalanalyses, microsensors, gene-basedidentification, microbial physiology techniquesand integrated modelling and simulation. Examples of the application of such techniquesare provided to demonstrate the way thesetechniques may be used in future. In the nextfew years, there is likely an exciting andhighly interactive period of research anddevelopment for the wastewater industry.     

Cloning and characterization of the hamster and guinea pig nicotinic acid receptors

In this study, we present the identification and characterization of hamster and guinea pig nicotinic acid receptors. The hamster receptor shares approximately 80-90% identity with the nucleotide and amino acid sequences of human, mouse, and rat receptors. The guinea pig receptor shares 76-80% identity with the nucleotide and amino acid sequences of these other species. [(3)H]nicotinic acid binding affinity at guinea pig and hamster receptors is similar to that in human (dissociation constant = 121 nM for guinea pig, 72 nM for hamster, and 74 nM for human), as are potencies of nicotinic acid analogs in competition binding studies. Inhibition of forskolin-stimulated cAMP production by nicotinic acid and related analogs is also similar to the activity in the human receptor. Analysis of mRNA tissue distribution for the hamster and guinea pig nicotinic acid receptors shows expression across a number of tissues, with higher expression in adipose, lung, skeletal muscle, spleen, testis, and ovary.     

Current and historical patterns of heavy metals pollution in Estonia as reflected in natural media of different ages: ICP Vegetation,ICP Forests and ICP Integrated Monitoring data

In order to characterise current and historical pattern of heavy metal (HM) pollution in Estonia, this article will compare the concentrations and stocks of Cd, Cr, Cu, Ni, Pb and Zn represented in current deposition (data from 18 local precipitation stations) with natural media of three different ages: 1–3-year-old moss carpet (ICP Vegetation moss survey data from 99 open area plots), 3–5-year-old litter layer, and several-decades-old organic layer (mor humus) of coniferous forest, in mostly podzolic soils (ICP Forest soil survey data, 75 stands).Objectives of this study are (1) to assess differences in HM retention and accumulation in various aged media of coniferous stands (2) to estimate territorial differences in current HM distribution and previously accumulated concentrations and stores of HM (3) to compare territorial distribution of HM concentration in Estonia between five different regions: N-W; N-E; S-W; S-E and Western insular region, whereas the local oil shale industry in N-E part of Estonia has been the main source of HM pollution over a long period of time and therefore may have an effect on HM regional distribution.Comparing the studied media, three types of HM retention patterns were detected: (1) for Cu, Ni, Cr (2) for Cd, Pb, (3) for Zn. The mean current level of HM deposition in Estonia is low comparison to previous decades, especially the 1980s. The effect of the previously significantly higher exposure of HM emissions and deposition is preserved in older part of soil organics (OF), where the highest stocks and concentrations of HMs (with the exception of Zn) are currently found. The HM proportions in fly ash of oil shale and in OF layer of soil were very similar with regards to Ni and Cr—indicating their origin from the oil shale industry in the N-E region. According to spatial distribution analysis, the greatest accumulated storages of Ni and Cr in OF layer of coniferous forest soils are characteristic to S-W Estonia.     

Confocal Raman microspectroscopy as a tool for studying the chemical heterogeneities of biofilms in situ

AIMS: To investigate the use of confocal Raman microspectroscopy (CRM) for the analysis of the structure, composition and development of fully hydrated biofilms. METHODS AND RESULTS: Pseudomonas aeruginosa PAO1 biofilms were cultured in a flow cell in minimal nutrient medium (artificial sea water) and their development was followed for up to 3 weeks. The spectroscopic signature of the biofilm cells and extracellular polymeric substances (EPS) were differentiated and their distribution in biofilm colonies and within water channels was mapped in-plane and -depth. The colonies were initially amorphous, mainly composed of cells with no detectable amount of EPS. They developed rapidly to give round colonies composed of a cellular core enclosed in a sheath of EPS. The EPS continued to increase and spread throughout the biofilm to become the dominating feature of aged colonies. Colonies with a liquid core morphology - characteristic of the seeding dispersal process - were also observed. CONCLUSIONS: This study demonstrated that CRM can be used to monitor the distribution of biofilm components in fully hydrated undisturbed biofilms over time. SIGNIFICANCE AND IMPACT OF THE STUDY: Confocal Raman microspectroscopy facilitates the analysis of hydrated, live bacterial biofilms as a function of space and time, thus making it a suitable technique for investigating the effects of various additives and environmental factors on biofilm growth.     

Phenylarsine oxide interferes with the death inducing signaling complex and inhibits tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induced apoptosis

The mechanism by which tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces death is the subject of intense scrutiny due to its preferential targeting of transformed cells for deletion. Based on recent findings that the TRAIL-dependent death inducing signaling complex (DISC) forms and signals at the plasma membrane without being internalized, we investigated the possibility that agents that prevent endocytosis may stabilize the surface bound DISC and thereby enhance TRAIL-dependent signaling. We utilized phenylarsine oxide (PAO), a trivalent arsenical that has been reported to inhibit endocytosis and to induce mitochondrial permeability transition. Therefore PAO could, by two separate and independent activities, enhance TRAIL-induced killing. Paradoxically, we found that rather than synergizing with TRAIL, PAO was an effective inhibitor of TRAIL-induced killing. Recruitment of FADD and caspase-8 to the TRAIL-dependent DISC was diminished in a concentration-dependent manner in cells exposed to PAO. The effects of PAO could not be reversed by washing cells under non-reducing conditions, suggesting covalent linkage of PAO with its cellular target(s); however, 2,3-dimercaptoethanol effectively overcame the inhibitory action of PAO and restored sensitivity to TRAIL-induced apoptosis. PAO inhibited formation of the TRAIL-dependent DISC and therefore prevented all subsequent apoptotic events.