Application of bioaffinity technology in therapeutic extracorporeal plasmapheresis and large-scale fractionation of human plasma

This paper describes the increasingly unique and powerful role that affinity chromatography is occupying both as a tool for the treatment of extracorporeal plasma exchange (to discard biological compounds with noxious metabolic or immunologic effects in patients) and as a purification tool in the production of therapeutic plasma protein derivatives. Management of both applications requires careful monitoring of the parameters applied to the plasma material, to avoid immunological stimulation or activation of the coagulation cascade. Examples of direct current applications of affinity ligands in therapeutic removal and industrial production of plasma compounds are presented.     

Natural evidence for chemical and early biological evolution

Meteorites, particularly type II carbonaceous chondrites, provide natural, tangible evidence for chemical evolution, but they do not appear to contain any evidence for biological evolution. On the other hand, some of the oldest sedimentary rocks of the earth have yielded good evidence for early biological evolution; whatever evidence there may be for chemical evolution in these old rocks is generally obscure. Carbonaceous chondrites (types I, II, and III) have been examined for thier content of various kinds of organic compounds. Amino acids have been reported to be present in the three types, but only in type II carbonaceous chondrites (Murray and Murchison) has an indigenous suite of amino acids been found which is apparently free of most terrestrial contaminations. These indigenous compounds are thought to have resulted from extraterrestrial, abiotic, chemical syntheses, and the presence of the amino acids in meteorites provides strong support for the theory of chemical evolution. The geological record of the Swaziland Sequence and Bulawayan System of Southern Africa contains morphological and chemical fossils which indicate that early biological evolution was taking place at least 3.0 to 3.3 aeons ago. Interpretation of the significance of the chemical fossil record has proven to be difficult. At present the occurrence of simple compounds in these very ancient rocks is believed to have little or nothing to do with biochemical processes three aeons ago. The bulk of the reduced carbonaceous material in these rocks, however, probably represents the residue of three billion years old and older organic matter. Isotopic studies of this carbonaceous material may provide chemical evidence for early biological evolution.     

Biofiltration of volatile organic compounds

The removal of volatile organic compounds (VOCs) from contaminated airstreams has become a major air pollution concern. Improvement of the biofiltration process commonly used for the removal of odorous compounds has led to a better control of key parameters, enabling the application of biofiltration to be extended also to the removal of VOCs. Moreover, biofiltration, which is based on the ability of micro-organisms to degrade a large variety of compounds, proves to be economical and environmentally viable. In a biofilter, the waste gas is forced to rise through a layer of packed porous material. Thus, pollutants contained in the gaseous effluent are oxidised or converted into biomass by the action of microorganisms previously fixed on the packing material. The biofiltration process is then based on two principal phenomena: (1) transfer of contaminants from the air to the water phase or support medium, (2) bioconversion of pollutants to biomass, metabolic end-products, or carbon dioxide and water. The diversity of biofiltration mechanisms and their interaction with the microflora mean that the biofilter is defined as a complex and structured ecosystem. As a result, in addition to operating conditions, research into the microbial ecology of biofilters is required in order better to optimise the management of such biological treatment systems.     

Reversible self-polymerizing high T(g) lyoprotectants

One mode of action of protectants in the storage of biological materials is by promoting the formation of a vitrified state on cooling or drying. In the case of preservation by drying, the glassy material comprises a low water content mixture of protectant and organic material. The protectant must on drying form a glassy state of glass transition temperature (T(g)) above the desired storage temperature. However, in some applications it must also be easily transported through cell membranes and this restricts the choice to a relatively limited number of small molecules, which typically exhibit very low glass transition temperatures. In this work we describe a self-polymerizing protectant comprising an inorganic salt and a small hydroxy functional molecule such as glycerol. This forms co-ordinate polymer chains of high T(g) on drying but rapidly depolymerizes into the original components on rehydration. The polymerization process is general for polyhydroxy compounds including glucose and related compounds.     

The organic stratigraphy of Ontong Java Plateau Tuff correlated with the depth‐related presence and absence of putative microbial alteration structures

Structures in geological samples are often interpreted as fossilised life; however, such interpretations are equivocal, as abiotic processes can be invoked to explain their presence. Thus, additional lines of chemical evidence are invaluable in confirming or refuting such morphological evidence. Glass shards in tuff from the Ontong Java Plateau (OJP) contain microtubular structures that are in close proximity to functionalised nitrogen substituted aromatic compounds that may be indicative of the chemical remnants of biological activity. The organic composition of the OJP tuff containing microtubular alteration structures was compared with tuff without such features. In addition, organic matter associated with horizons with compacted remnants of woody material buried in the OJP tuff and overlying pelagic calcareous foraminifer sediment were also characterised, to ascertain the provenance of the organic matter found in the OJP tuff. As a further control, the organic material in submarine and terrestrial basalts from other locations were also characterised providing further evidence to support the view that the organic matter in the OJP tuff is authigenic. Carbon–nitrogen chemistry was detected across all OJP tuff samples irrespective of the presence or absence of microtubular features, but was not detected in either the wood material, the overlying pelagic sediments or in the basalts from other locations. The results indicate no direct link between the OJP nitrogenous organic compounds and the presence or absence of microtubular features.     

Direct detection of proteins adsorbed on synthetic materials by matrix-assisted laser desorption ionization-mass spectrometry.

The irreversible accumulation of biological material on synthetic surfaces ("biofouling") adversely affects for instance contact lenses, implantable biomedical devices, biosensors, water purification, transport and storage systems, and marine structures. It is shown here that proteins adsorbed on contact lenses can be detected directly, rapidly, and conveniently, with high sensitivity, by matrix-assisted laser desorption ionization (MALDI)-mass spectrometry. This new approach allows detection of minor (and major) proteinaceous constituents of biofouled layers on samples retrieved from clinical usage and in vitro protein adsorption studies, at levels substantially below monolayer coverage. Identification of the detected biological molecules can be done by comparison of the detected mass peaks with known protein molecular masses or with spectra recorded of pure compounds or by separate biochemical assays. The MALDI mass spectra recorded on different contact lenses contain peaks assignable to lysozyme and a number of smaller proteins. Such sensitive characterization of the early stages of biofouling enhances the understanding of protein/materials interactions and assists in designing guided strategies toward control of biological adsorption processes.     

A synthetic approach to novel carvotacetone and antheminone analogues with anti-tumour activity

A synthetic approach to analogues of the terpenoid natural product antheminone A is described which employs (?)-quinic acid as starting material. A key conjugate addition step proved to be unpredictable regarding its stereochemical outcome however the route allowed access to two diastereoisomeric series of compounds. The results of biological assay of the toxicity of the target compounds towards non-small-cell lung cancer cell line A549 are reported.     

Comparison of two methods used to analyse lipid peroxidation from Vaccinium myrtillus (L.) during snow removal, reacclimation and cold acclimation.

Malondialdehyde (MDA) concentration is a widely used method to analyse lipid peroxidation in biological material. In plant tissues, however, certain compounds (anthocyanins, carbohydrates) may interfere with measurements which may lead to an overestimation of the MDA levels. Two methods were compared for analysing lipid peroxidation, either uncorrected or corrected for interfering compounds. The comparison was performed in three separate experiments with respect to cold treatments (snow removal in winter, reacclimation in summer and cold acclimation in autumn) in bilberry (Vaccinium myrtillus L.). During winter and autumn the methods seem to measure different compounds, but during active growth in the summer the difference between the methods was less. This is obviously due to carbohydrates which act as cryoprotectants and increase in concentration during cold acclimation as well as due to the anthocyanins. It is thus suggested that the validity of the uncorrected method to measure MDA and thereby lipid peroxidation is best in plant tissue which is in an active growth state.     

Structural and Biological Diversity of Cyclic Octadecanoids,Jasmonates, and Mimetics

The jasmonate family of plant signaling compounds comprises biologically highly active cyclopentenones (for example, 12-oxo-phytodienoic acid) (12-OPDA) and cyclopentanones (for example, jasmonic acid) (JA) of related origin via the octadecanoid pathway, and structure. Among others, their biological activities include a broad range of defense-related reactions. Several lines of evidence indicate both common and different biological responses mediated by 12-OPDA and/or JA, suggesting the existence of at least two separate structure-activity groups. Based on the structure of a bacterial phytotoxin, coronatine, with similar biological activities compared with jasmonates, indanoyl isoleucine conjugates have been designed as functional synthetic mimics of octadecanoid-derived signals. The structural diversity of naturally occurring jasmonate-related compounds and synthetic mimics is discussed with respect to their corresponding biological activities. Novel strategies for the synthesis of various indanoyl isoleucine conjugates will be presented.     

Tritium labeling of antisense oligonucleotides by exchange with tritiated water.

We describe a simple, efficient, procedure for labeling oligonucleotides to high specific activity (< 1 x 10(8) cpm/mumol) by hydrogen exchange with tritiated water at the C8 positions of purines in the presence of beta-mercaptoethanol, an effective radical scavenger. Approximately 90% of the starting material is recovered as intact, labeled oligonucleotide. The radiolabeled compounds are stable in biological systems; greater than 90% of the specific activity is retained after 72 hr incubation at 37 degrees C in serum-containing media. Data obtained from in vitro cellular uptake experiments using oligonucleotides labeled by this method are similar to those obtained using 35S or 14C-labeled compounds. Because this protocol is solely dependent upon the existence of purine residues, it should be useful for radiolabeling modified as well as unmodified phosphodiester oligonucleotides.     

Chemical reactivity and mutagenicity of some dihalomethanes

Four dihalomethanes; dichloromethane, bromochloromethane, dibromomethane and diiodomethane, have been studied with respect to their reactivities towards nucleophilic compounds of different strengths in water solution and with respect to their toxicities and mutagenic effectiveness in bacterial test systems. The correlation between biological activity (toxicity and mutagenic effectiveness in Salmonella TA100) and reactivity towards strong nucleophiles indicates that reactions with nucleophilic groups of high reactivity in the biological material, possibly SH or amino groups in proteins, are involved in their mechanism of action.     

Evolution of Volatile Sulfur Compounds during Laboratory-Scale Incubations and Indoor Preparation of Compost Used as a Substrate in Mushroom Cultivation

Volatile sulfur compounds are known to be produced during the preparation of compost used as a substrate in mushroom cultivation. Because they cause odor problems, attempts have been made to reduce the production of these compounds. The influences of temperature and various additions on the production of volatile sulfur compounds from composting material were tested on laboratory-scale preparations. The production of H₂S, COS, CH₃SH, and (CH₃)₂S was proven to be a biological process with an optimal temperature that coincides with the optimal temperature for biological activity. The formation of CS₂ and (CH₃)₂S₂ was shown to be a nonbiological process. The emission of volatile sulfur compounds during the indoor preparation of mushroom compost appeared to be remarkably reduced (about 90%) as compared with the emission during the conventional outdoor process. Introduction of this indoor composting process would result in a significant reduction in environmental pollution.     

Repellent and Insecticidal Activity of Naphthoquinones from the Heartwood of <i>Tectona grandis</i> on <i>Incisitermes marginipennis</i> (Latreille)

Wood is a lignocellulosic material that, because of its organic nature, is a source of nutrients for bacteria, fungi, and insects. Incisitermes marginipennis (Latreille) is an interesting dry wood-boring termite. Because it is an extremely destructive pest, difficult to control due to its cryptic lifestyle, and for its role in the biogeochemical cycle of carbon and other elements; it is a fundamental organism. The use of less toxic and eco-friendly substances is important for protecting domestic, cultural, and historical heritage made of wood or lignocellulosic materials. The aim of this study was to evaluate the biological activity of various naphthoquinones extracted from the heartwood of T. grandis on I. marginipennis. First, heartwood flour was used for Soxhlet-type extractions to isolate and characterize the heartwood compounds by thin-layer chromatography. To identify the heartwood compounds, nuclear magnetic resonance and gas chromatography coupled to a mass spectrometer were used. The information obtained was supported using the Interactive Spectroscopy software. The identified heartwood compounds were lapachol, tectoquinone, deoxylapachol, dehydro-α-lapachone, tectol and tecomaquinone-I. Using nymphal termites of I. marginipennis as a biological model, the isolated and identified molecules were under three different tests (repellent, insecticide and protective) and their biological activity was individually evaluated. The results showed that the molecules obtained have a repellent-protective effect against I. marginipennis, although not in all cases, these molecules have high percentages of mortality and decreased the weight loss of the treated material exposed to I. marginipennis.     

Microwave-assisted extraction,HPLC analysis,and inhibitory effects on carbonic anhydrase I,II, VA,and VII isoforms of 14 blueberry Italian cultivars

Abstract

The multi-component fingerprint and the biological evaluation of plant-derived material are indispensable for the pharmaceutical field, in food quality control procedures, and in all plant-based products. We investigated the quantitative content of biologically active compounds (anthocyanins and chlorogenic acid) of microwave-assisted blueberry extracts from 14 different Italian cultivars, using validated high-performance liquid chromatography-photodiode array detector (HPLC-PDA) method and routinely instrument configuration. The carbonic anhydrase (CA, EC 4.2.1.1) inhibition profiles against several pharmacologically relevant CA isoforms of blueberry extracts and some bioactive compounds were also investigated. The various cultivars showed a highly variable content in anthocyanins and chlorogenic acid, and their CA inhibitory effects were also highly variable. Overall these data prove that antioxidant natural products found in blueberries may be useful for designing pharmacological agents in which various CAs are involved, e.g., antiobesity, antitumor, or anticonvulsants agents.     

Synthesis, characterization and reactions of 2-deoxo-5-deazaalloxazines

5-Deazaflavins and their homologues have been known as potential riboflavin antagonists, bioreductives, and compounds with potent antitumor activity. 2-Amino-4-methylquinoline-3-carbonitrile (2) was prepared as unreported starting material for several interesting 2-deoxo-5-deazalloxazine derivatives. Cyclization of 2 using formamide afforded the 2,4-deoxo-5-deazaalloxazine derivative 7, which was subjected to deamination with nitrous acid to give the 2-deoxo-5-deazaalloxazine (8). The compound 8 was also obtained via 13 by treating the latter with refluxing formic acid or formamide and used as a precursor for synthesis of several 2-deoxo-5-deazaalloxazines 18, 19, 20, 21 and 22. The pharmacological and biological properties of these compounds are still under investigation.     

Distinctive hydrocarbons of the black dump fly, Hydrotaea aenescens (Diptera: Muscidae).

Hydrotaea aenescens (Wiedemann), the black dump fly, is a potential biological control agent originally from the western hemisphere, now found in many parts of the Palearctic region except for the United Kingdom, where it cannot be imported for any reason. A complication of classical biological control is the problem of strain identification, as one must be able to somehow mark or follow a particular strain that has been introduced into the field or is contemplated for release. Gas chromatographic analysis of the surface hydrocarbons of pooled and individual dump fly adults resulted in reproducible hydrocarbon patterns that differentiated widely distributed strains of H. aenescens and showed similarities between strains that were related. Sexual dimorphism was observed in the surface hydrocarbons. Conspecific similarities included identities of the hydrocarbons found in colony material collected worldwide, with differences being found in the quantities of compounds present.     

Rediscovery of known natural compounds: nuisance or goldmine?

Do all natural compounds have a distinct biological activity, or are most of them merely biosynthetic debris? Many natural compounds have important biological functions, and certainly many more of the ample 200,000 currently known will ultimately prove to be more than just 'secondary metabolites'. The question is how to select the most promising candidates for potential new drugs. 'Rediscovery' of known natural compounds is regarded as a nuisance or disappointment by scientists involved with the identification of novel compounds. The other side of the coin, however, is that the discovery that a particular compound occurs in unrelated species can be a valuable clue toward the identification of a novel receptor or enzyme. Here, we put forward the hypothesis that when a natural compound occurs in unrelated species, it must have an important biological function by interacting with a specific molecular target. This is because it is extremely improbable that in nature one particular compound is synthesized in totally unrelated species for no reason at all. For many compounds occurring in unrelated species, it is already known that they act on specific molecular targets. For others, it is just known that they occur in different species. In some cases, biological activities are known but not the underlying mechanisms of action. It is from this category of compounds that important discoveries are likely to be made. Some (around 70) of them were identified. They represent important clues from nature offering an alternative approach to the classical screening of large numbers of compounds.     

The determination of S-nitrosothiols in biological samples--procedures, problems and precautions

Despite the considerable number of published studies in the field of S-nitrosothiols (RSNO), the determination of these compounds in biological samples still represents an analytical challenge, due to several technical obstacles and often long sample preparation procedures. Other problems derive from the intrinsic lability of RSNO and the absence of certified reference material, analytically validated methods or suitable internal standards. Also, thiols and nitrites are usually present at high concentrations in biological matrices, and all precautions must be adopted in order to prevent artifactual formation of RSNO. Preanalytical steps (sampling, preservation and pre-treatment of samples) are particularly critical for the obtainment of reliable measurements. Three main mechanisms have been identified capable of compromising the assays: metal-catalyzed RSNO decomposition, reduction of the S-NO bond by thiols (transnitrosylation reactions) and enzymatic degradation of S-nitroso-glutathione (GSNO) by endogenous γ-glutamyltransferase (GGT) activity possibly present in the sample. If not adequately controlled, these factors likely contribute to the wide dispersion of values reported in the literature for RSNO and GSNO concentration in biological fluids, blood in the first place. The use of metal chelators, thiol reagents and GGT inhibitors appears therefore mandatory.     

Processing scarce biological samples for light and transmission electron microscopy

Light microscopy (LM) and transmission electron microscopy (TEM) aim at understanding the relationship structure-function. With advances in biology, isolation and purification of scarce populations of cells or subcellular structures may not lead to enough biological material, for processing for LM and TEM. A protocol for preparation of scarce biological samples is presented. It is based on pre-embedding the biological samples, suspensions or pellets, in bovine serum albumin (BSA) and bis-acrylamide (BA), cross-linked and polymerized. This preparation provides a simple and reproducible technique to process biological materials, present in limited quantities that can not be amplified, for light and transmission electron microscopy.