Cartilage oligomeric matrix protein deficiency promotes early onset and the chronic development of collagen-induced arthritis

Introduction  

Cartilage oligomeric matrix protein (COMP) is a homopentameric protein in cartilage. The development of arthritis, like collagen-induced arthritis (CIA), involves cartilage as a target tissue. We have investigated the development of CIA in COMP-deficient mice.     

Water solubilization of DDGS via derivatization with phosphite esters

Ethanol production from corn starch in the corn dry milling process leaves Distillers' Dry Grains and Solubles (DDGS) as a major by-product from which additional ethanol may be economically obtained from its glucan content. A challenge in processing the cellulose content of this material lies in its extensive inter-cellulose chain hydrogen bonding, which inhibits access of enzymes capable of cleaving glycosidic bonds, a transformation required for providing fermentable sugars. The phosphitylation of cellulosic OH groups using a reactive bicyclic phosphite ester is utilized to disrupt cellulosic hydrogen bonds, thus providing access to cellulose chains for further processing. We describe a method of pretreating DDGS with commercially available trimethylolpropane phosphite [P(OCH2)3CEt] in the presence of a slight molar excess of water to afford greater than 90% DDGS solubility in the reaction mixture in methanol and in water. Preliminary results using a model compound [D-(+)-permethylated cellobiose] indicate that glycosidic bonds are cleaved as a consequence of this pretreatment.     

Purification from human milk of matriptase complexes with secreted serpins: mechanism for inhibition of matriptase other than HAI-1

Matriptase, a type 2 transmembrane serine protease, is predominately expressed by epithelial and carcinoma cells in which hepatocyte growth factor activator inhibitor 1 (HAI-1), a membrane-bound, Kunitz-type serine protease inhibitor, is also expressed. HAI-1 plays dual roles in the regulation of matriptase, as a conventional protease inhibitor and as a factor required for zymogen activation of matriptase. As a consequence, activation of matriptase is immediately followed by HAI-1-mediated inhibition, with the activated matriptase being sequestered into HAI-1 complexes. Matriptase is also expressed by peripheral blood leukocytes, such as monocytes and macrophages; however, in contrast to epithelial cells, monocytes and macrophages were reported not to express HAI-1, suggesting that these leukocytes possess alternate, HAI-1-independent mechanisms regulating the zymogen activation and protease inhibition of matriptase. In the present study, we characterized matriptase complexes of 110 kDa in human milk, which contained no HAI-1 and resisted dissociation in boiling SDS in the absence of reducing agents. These complexes were further purified and dissociated into 80-kDa and 45-kDa fragments by treatment with reducing agents. Proteomic and immunological methods identified the 45-kDa fragment as the noncatalytic domains of matriptase and the 80-kDa fragment as the matriptase serine protease domain covalently linked to one of three different secreted serpin inhibitors: antithrombin III, 1-antitrypsin, and 2-antiplasmin. Identification of matriptase-serpin inhibitor complexes provides evidence for the first time that the proteolytic activity of matriptase, from those cells that express no or low levels of HAI-1, may be controlled by secreted serpins. protease; type 2 transmembrane serine protease; protease inhibitor; ST-14; hepatocyte growth factor activator inhibitor 1     

Integrated flow-injection processing for on-line quantification of plasmid DNA during cultivation of E. coli

An integrated flow-injection processing (FIP) system for the quantification of plasmids during cultivation is described. The system performs on-line sampling, cell lysis, and quantification of plasmids in an integrated manner during cultivation of E. coli. The system was operated by using a miniaturized expanded-bed column which can be used for handling samples containing cells and cell debris without interfering with the binding analysis. Two types of detectors (one measuring UV absorbance at 254 nm and a fluorometer) are used for on-line plasmid detection. The system was developed using standard solutions and it was successfully applied in monitoring plasmid contents during a cultivation of E. coli.     

In vitro laser ablation of natural marine biofilms

We studied the efficiency of pulsed low-power laser irradiation of 532 nm from an Nd:YAG (neodymium-doped yttrium-aluminum-garnet) laser to remove marine biofilm developed on titanium and glass coupons. Natural biofilms with thicknesses of 79.4 +/- 27.8 microm (titanium) and 107.4 +/- 28.5 microm (glass) were completely disrupted by 30 s of laser irradiation (fluence, 0.1 J/cm2). Laser irradiation significantly reduced the number of diatoms and bacteria in the biofilm (paired t test; P < 0.05). The removal was better on titanium than on glass coupons.     

Factors Affecting the Agrobacterium-Mediated Transient Transformation of the Wetland Monocot,Typha latifolia

An Agrobacterium-mediated transformation system, using transient transformation assays, was used to evaluate conditions influencing transformation for the wetland monocot Typha latifolia. These studies were aimed at the long-term objective of evaluating candidate genes for phytoremediation. The binary plasmid vector pCAMBIA1301/EHA105, containing the -glucuronidase coding sequence, was used in combination with factors known to affect transformation. These included callus age at the time of cocultivation with Agrobacterium tumefaciens, type and concentration of auxin for explant growth, light or dark culture environment, the presence or absence of acetosyringone (AS), explant type, explant wounding and the number of days used for cocultivation. The number of days needed for the first detection of transient expression of the -glucuronidase gene was also examined. Three days of Agrobacterium cocultivation of 50-day-old seedling-derived calluses, grown on 20.7 µM (5 mg l^–1) picloram supplemented medium, in the dark, resulted in higher levels of transient -glucuronidase expression than were seen in calluses cultured on 4.5 or 22.6 µM (1 or 5 mg l^–1) 2,4-dichlorphenoxyacetic acid containing media. The addition of 100 µM acetosyringone significantly enhanced transient -glucuronidase activity. Wounding of explants, by cutting into two or three pieces, 3 days before cocultivation, increased expression of -glucuronidase only in calluses cultured under light conditions. Transient -glucuronidase expression was observed as early as 24 h after cocultivation and increased as the days post cultivation increased. The developed transient system will be used for stable transformation of Typha species.     

Limited microsynteny between the genomes of Pristionchus pacificus and Caenorhabditis elegans

Nematodes are an attractive group of organisms for studying the evolution of developmental processes. Pristionchus pacificus was established as a satellite organism for comparing vulva development and other processes to Caenorhabditis elegans. The generation of a genetic linkage map of P.pacificus has provided a first insight into the structure and organization of the genome of this species. Pristionchus pacificus and C.elegans are separated from one another by >100 000 000 years such that the structure of the genomes of these two nematodes might differ substantially. To evaluate the amount of synteny between the two genomes, we have obtained 126 kb of continuous genomic sequence of P.pacificus, flanking the developmental patterning gene pal-1. Of the 20 predicted open reading frames in this interval, 11 have C.elegans orthologs. Ten of these 11 orthologs are located on C.elegans chromosome III, indicating the existence of synteny. However, most of these genes are distributed over a 12 Mb interval of the C.elegans genome and only three pairs of genes show microsynteny. Thus, intrachromosomal rearrange ments occur frequently in nematodes, limiting the likelihood of identifying orthologous genes of P.pacificus and C.elegans based on positional information within the two genomes.     

Laser impact on bacterial ATP: insights into the mechanism of laser-bacteria interactions

The mechanisms of laser action on bacteria are not adequately understood. Here, an attempt has been made to study the fluctuation in ATP (adenosine triphosphate) concentration following laser irradiation from a pulsed Nd:YAG laser on a marine biofilm-forming bacterium Pseudoalteromonas carrageenovora. A stationary phase bacterial suspension (density 10(7-8) ml-1) was exposed to pulsed laser irradiations at a fluence of 0.1 J cm-2 (pulse width 5 ns, repetition rate 10 Hz) for different durations, ranging from 2 s to 15 min. The total viable count (TVC) and ATP concentration of the irradiated samples were determined immediately after the laser irradiation. While the maximum reduction in the TVC observed with respect to the control was 59% immediately after 15 min irradiation, the ATP concentration showed a reduction of about 86% for the same duration. The ATP concentration showed an abrupt reduction from 3 min of laser irradiation and continued to reduce significantly with increasing duration of irradiation. Thus, 3 min irradiation at a fluence of 0.1 J cm-2 is considered as an approximate threshold for ATP production in this bacterium. As the decreased level of ATP production continued, bacterial mortality resulted. The reduction in ATP production could be due to damage caused by the laser irradiations on bacterial metabolic processes such as cellular respiration.     

Microfouling studies on experimental test blocks of steel-making slag and concrete exposed to seawater off Chiba, Japan

Microfouling studies with the emphasis on microalgae (Bacillariophyceae) were carried out on test blocks of steel-making slag in comparison with concrete. Two types of slag test blocks, with and without fly-ash as an additional source of silica, and concrete test blocks of size 75 x 26 x 26 mm were used to study microfouling build-up for a period of 30 d, with intermittent samplings after 1, 2, 3, 7, 14 and 21 d. The species composition, cell density, biomass and surface pH of the test pieces were determined, in addition to the hydrographic parameters of the water column. Microfouling studies showed higher numbers of algal species as well as a greater cell density on the slag than on the concrete blocks. This was true with respect to biomass measured as dry weight also. Colonization was significantly delayed in the case of concrete. Navicula spp. and Nitzschia spp. were the initial colonizers on all three types of substrata and were the dominant genera throughout the study period. While the number of species increased, several disappeared after colonization, as a part of community build-up. The surface pH of the slag blocks was near neutral, whilst that of the concrete was highly alkaline during the initial period of exposure. This alkaline surface reduced the rate of species colonization on the concrete blocks initially. The study showed severe biofouling on the slag blocks compared to concrete and thus they were considered an environmentally benign construction material for land protection. The use of slag as the construction material for land protection would greatly reduce the expense compared to concrete.     

Bacterial destruction of mica during bioleaching of kaolin and quartz sandsby Bacillus cereus

Growth and metabolic activities of Bacillus cereus were found to cause the extraction of iron atoms from the octahedral position in mica in the kaolin sample (49%) and in the quartz sands sample (17%) after 3 months of bioleaching, while aluminium removal was only 5%. Mica destruction was detected in kaolin and quartz sands samples by X-ray diffraction analysis and also by i.r. adsorption spectroscopy in quartz sands samples. The structural changes obtained were confirmed by scanning electron microscopy (SEM) analysis. The SEM pictures show a different morphology in the boundary region of mica grains before and after bioleaching. Bacterial destruction effects were feeble in the interlayer sites and were specially directed to split planes, which are occupied by a number of bacterial cells. The biological destruction of mica with phengite composition after iron removal led to development of illite, which was detected by energy-dispersion microanalysis (EDS). Illite development caused also the enrichment of the kaolin sample by fine-grained fraction.