Synthesis and bioevaluation of some new isoniazid derivatives

The aim of the study was to synthesize some new compounds with potential anti-tuberculosis activity, containing isoniazid and α,β-unsaturated thiocinnamamide-like thioamides as precursors. The obtained derivatives were evaluated regarding their biological activity (antioxidant and antibacterial), as well as their influence on the eukaryotic cell cycle. The results suggested that the newly obtained derivatives of isoniazid exhibited different biological activities, depending on their structure; thus, the most active compound in terms of anti-oxidant and anti-Mycobacterium tuberculosis effects proved to be the isonicotinic acid N′-(1-amino-1-mercapto-3-phenyl-propen-1-yl)-hydrazide. This compound also increased the expression of NAT1 and NAT2 genes, which are implicated in the metabolism of the isoniazid, demonstrating that it could be rapidly metabolized, and thus well tolerated. The largest spectrum of antibacterial activity (excluding M. tuberculosis) was noticed for the isonicotinic acid N′-[1-amino-1-mercapto-3-(p-chloro-phenyl)-propen-1-yl]-hydrazide, which was also the most cytotoxic, especially at high concentrations, although not significantly affecting the cellular cycle phases. The obtained results showed that the new derivatives could represent potential candidates for the treatment of M. tuberculosis infections, but further research is needed in order to improve their pharmacological properties, by increasing their antimicrobial activity and reducing the risk of side-effects.     

Trichomonas vaginalis--identification by rapid BPT-Dr?gan staining

The blue-polychrome-tannin rapid staining (BPT-Dr?gan, 1968) applied on fresh and dried vaginal smears makes possible the identification of different forms of Trichomonas vaginalis (TV) parasite. The nucleus is coloured in blue-violet and the cytoplasm in various degrees of blue-green, violet or bichromatical, with blue and violet zones at the cell periphery. In the cytoplasm, the staining allows the identification of many granules and vacuoles. All the stainable substrates have an acid character and represent morphohistochemical aspects which reflect the parasite metabolism in different functional and evolutive stages. Our results allow the recommendation of the method in the screening actions of the TV identifications.     

Important Plant Areas: revised selection criteria for a global approach to plant conservation

Despite the severe threats to plant habitats and high levels of extinction risk for plant species in many parts of the world, plant conservation priorities are often poorly represented in national and global frameworks because of a lack of data in an accessible and consistent format to inform conservation decision making. The Important Plant Areas (IPAs) criteria system offers a pragmatic yet scientifically rigorous means of delivering these datasets, enabling informed national- or regional-scale conservation prioritisation, and contributing significantly towards global prioritisation systems including the International Union for Conservation of Nature Key Biodiversity Areas (KBAs) Standard. In this paper, we review the IPA rationale and progress on IPA identification to date, including the perceived limitations of the process and how these may be overcome. We then present a revised set of criteria for use globally, developed through the combined experiences of IPA identification over the past decade and a half and through a recent global consultation process. An overview of how the revised IPA criteria can work alongside the newly published KBA Standard is also provided. IPA criteria are based around a sound, scientific, global framework which acknowledges the practical problems of gathering plant and habitat data in many regions of the world, and recognises the role of peer reviewed expert opinion in the selection process. National and sub-national engagement in IPA identification is essential, providing a primary route towards long term conservation of key sites for plant diversity. The IPA criteria can be applied to the conservation of all organism groups within the plant and fungal kingdoms.     

High Prevalence of Multistability of Rest States and Bursting in a Database of a Model Neuron

Flexibility in neuronal circuits has its roots in the dynamical richness of their neurons. Depending on their membrane properties single neurons can produce a plethora of activity regimes including silence, spiking and bursting. What is less appreciated is that these regimes can coexist with each other so that a transient stimulus can cause persistent change in the activity of a given neuron. Such multistability of the neuronal dynamics has been shown in a variety of neurons under different modulatory conditions. It can play either a functional role or present a substrate for dynamical diseases. We considered a database of an isolated leech heart interneuron model that can display silent, tonic spiking and bursting regimes. We analyzed only the cases of endogenous bursters producing functional half-center oscillators (HCOs). Using a one parameter (the leak conductance ()) bifurcation analysis, we extended the database to include silent regimes (stationary states) and systematically classified cases for the coexistence of silent and bursting regimes. We showed that different cases could exhibit two stable depolarized stationary states and two hyperpolarized stationary states in addition to various spiking and bursting regimes. We analyzed all cases of endogenous bursters and found that 18% of the cases were multistable, exhibiting coexistences of stationary states and bursting. Moreover, 91% of the cases exhibited multistability in some range of . We also explored HCOs built of multistable neuron cases with coexisting stationary states and a bursting regime. In 96% of cases analyzed, the HCOs resumed normal alternating bursting after one of the neurons was reset to a stationary state, proving themselves robust against this perturbation.     

The biogenesis protein PEX14 is an optimal marker for the identification and localization of peroxisomes in different cell types, tissues, and species in morphological studies

Catalase and ABCD3 are frequently used as markers for the localization of peroxisomes in morphological experiments. Their abundance, however, is highly dependent on metabolic demands, reducing the validity of analyses of peroxisomal abundance and distribution based solely on these proteins. We therefore attempted to find a protein which can be used as an optimal marker for peroxisomes in a variety of species, tissues, cell types and also experimental designs, independently of peroxisomal metabolism. We found that the biogenesis protein peroxin 14 (PEX14) is present in comparable amounts in the membranes of every peroxisome and is optimally suited for immunoblotting, immunohistochemistry, immunofluorescence, and immunoelectron microscopy. Using antibodies against PEX14, we could visualize peroxisomes with almost undetectable catalase content in various mammalian tissue sections (submandibular and adrenal gland, kidney, testis, ovary, brain, and pancreas from mouse, cat, baboon, and human) and cell cultures (primary cells and cell lines). Peroxisome labeling with catalase often showed a similar tissue distribution to the mitochondrial enzyme mitochondrial superoxide dismutase (both responsible for the degradation of reactive oxygen species), whereas ABCD3 exhibited a distinct labeling only in cells involved in lipid metabolism. We increased the sensitivity of our methods by using QuantumDots?, which have higher emission yields compared to classic fluorochromes and are unsusceptible to photobleaching, thereby allowing more exact quantification without artificial mistakes due to heterogeneity of individual peroxisomes. We conclude that PEX14 is indeed the best marker for labeling of peroxisomes in a variety of tissues and cell types in a consistent fashion for comparative morphometry.     

Role of bicarbonate as a pH buffer and electron sink in microbial dechlorination of chloroethenes

ABSTRACT: BACKGROUND: Buffering to achieve pH control is crucial for successful trichloroethene (TCE) anaerobic bioremediation. Bicarbonate (HCO3-) is the natural buffer in groundwater and the buffer of choice in the laboratory and at contaminated sites undergoing biological treatment with organohalide respiring microorganisms. However, HCO3- also serves as the electron acceptor for hydrogenotrophic methanogens and hydrogenotrophic homoacetogens, two microbial groups competing with organohalide respirers for hydrogen (H2). We studied the effect of HCO3- as a buffering agent and the effect of HCO3--consuming reactions in a range of concentrations (2.5-30 mM) with an initial pH of 7.5 in H2-fed TCE reductively dechlorinating communities containing Dehalococcoides, hydrogenotrophic methanogens, and hydrogenotrophic homoacetogens. RESULTS: Rate differences in TCE dechlorination were observed as a result of added varying HCO3- concentrations due to H2-fed electrons channeled towards methanogenesis and homoacetogenesis and pH increases (up to 8.7) from biological HCO3- consumption. Significantly faster dechlorination rates were noted at all HCO3- concentrations tested when the pH buffering was improved by providing 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) as an additional buffer. Electron balances and quantitative PCR revealed that methanogenesis was the main electron sink when the initial HCO3- concentrations were 2.5 and 5 mM, while homoacetogenesis was the dominant process and sink when 10 and 30 mM HCO3- were provided initially. CONCLUSIONS: Our study reveals that HCO3- is an important variable for bioremediation of chloroethenes as it has a prominent role as an electron acceptor for methanogenesis and homoacetogenesis. It also illustrates the changes in rates and extent of reductive dechlorination resulting from the combined effect of electron donor competition stimulated by HCO3- and the changes in pH exerted by methanogens and homoacetogens.