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Distribution of esterase activity at the level of the epithelium of the diffusely infiltrated area (DIA) and of the cloaca in the Gallus domesticus: an ultrastructural study.

Esterase activity studies on the areas of lymphoid infiltration in the bursa of Fabricius (diffusely infiltrated area) and the dorsal wall of the cloaca showed that the epithelial cells exhibit varying degrees of diffuse esterase activity; it was possible to demonstrate the presence of spot-like esterase positivity in star-shaped cells in the epithelium itself. These cells can be found at various levels from the proximal to the distal region and have also been observed in the connective tissue of the tunica propria; as a result, the hypothesis has been advanced that they are connective tissue.

Animals↗

A dot immunobinding assay (DIA) on nitrocellulose membrane for the serological detection of antibodies to Entamoeba histolytica.

A rapid, cheap, simple and specific serological test of adequate sensitivity for detecting IgG antibodies against Entamoeba histolytica antigen is described. Axenically cultured amoebic antigen was used to precoat the nitrocellulose membrane. The strips were incubated with test samples and later with horseradish peroxidase (HRPO) labelled protein-A conjugate. A dark blue spot was obtained by treatment with peroxidase substrate, 4-chloro-1-naphthol, in positive samples. Serum samples from 32 healthy controls, 45 patients with acute amoebic liver abscess and 10 asymptomatic E. histolytica cyst passers were tested. This test was positive in 93% of cases of amoebic liver abscess, 3% of healthy controls and none of the cyst passers; its sensitivity (97%) and specificity (93%) were as good as that of the ELISA test and, because it is simple, quick and cheap it is recommended as a serological test of choice for the diagnosis of invasive amoebiasis.

Antigens, Protozoan↗

Digital image analysis (DIA) of colour changes in human skin exposed to standardized thermal injury and comparison with laser Doppler measurements.

Clinical macrophotography followed by digitization and computer-assisted image analysis was performed on volunteers exposed to a mild experimental thermal injury. The aim was to elaborate a non-invasive technique allowing repetitive and quantitative monitoring of induced erythema and to evaluate a possible relationship with laser Doppler measurement of skin perfusion. A standardized 1 x 1 cm large thermal trauma was induced in the skin of the ventral forearm of 12 volunteers. Photographic recordings and skin blood flow measurements were made preburn and at 30-min, 1-h, 2-h, 4-h and 12-h postburn. Image analysis was performed with two colour systems, normalized rgb-values (rgb) and Hue-Saturation-Intensity (HSI). Erythema measured by rgb and HSI was most pronounced during the first hour postburn, after which it gradually decreased in order to increase again at 12 h postburn. Skin perfusion peaked at 30 min postburn and then continuously decreased during the course of the experiments. Degree of erythema could be quantified using both colour systems and a linear relationship was obtained between the observed colour changes of both systems and changes in skin perfusion. Results may also indicate that erythema can be present without a concomitant increase in skin perfusion, probably representing postburn venous stasis. The present study showed that digital image analysis is a non-invasive technique allowing repetitive and quantitative analysis of skin erythema which can be used to monitor pathophysiological changes in the body as well as the result of their treatment.

Adult↗

Limited Impact of Column Chemistry and Length on Proteome Coverage Under High-Speed DIA.

The evolution of mass spectrometry (MS)-based proteomics has been driven by continuous technological advances in sample preparation, liquid-phase separations, instrumentation, and data acquisition. Chromatographic performance has been recognized as a contributing factor to identification depth, particularly on earlier-generation MS platforms. Recent advances in MS sampling speed and sensitivity now raise the question of how strongly chromatographic quality continues to determine overall proteome coverage. We investigate how column chemistry and length influence proteome coverage and chromatographic selectivity under modern data-independent acquisition conditions, and whether traditional optimization priorities still apply. Spanning a matrix of experiments with five distinct stationary phases, including C18 chemistries, C8, and Phenyl-Hexyl, across eight column lengths (40-140 mm), we evaluate protein identification performance using data-independent acquisition on the Orbitrap Astral mass spectrometer. Despite differences in stationary-phase chemistry and column length, we observed remarkably convergent proteome coverage metrics. All C18 and C8 phases consistently achieved over 150,000 precursor- and approximately 9000 protein group identifications, regardless of column length variations. While retention fingerprints persisted across chemistries, these chromatographic differences did not translate into meaningful variations in proteome coverage under high-speed acquisition conditions at 200 Hz. Within the range of modern sub-2 μm reversed-phase materials tested, identification depth showed limited dependence on column chemistry and length, suggesting that for state-of-the-art stationary phases, method development priorities may increasingly favor operational robustness, throughput, and reproducibility over traditional separation optimization.

Proteome↗

Prevalence of diego blood group Dia antigen in Mongolians: comparison with that in Japanese.

The Diego blood group is composed of Di(a) and Di(b) antigens. Prevalence of the Di(a) antigen is known to be different among races. The Di(a) antigen is generally found in Oriental people. Thus, it is called a Mongoloid factor. In Japanese, the prevalence of this antigen is 8.78%. However, the prevalence in Mongolians had not previously been examined. In September of 2002, we determined this antigen among inhabitants of Ulaanbaatar. It was found in 24 of 242 subjects (9.92%). This prevalence approximates that in Japanese. The Rh blood group phenotypes also showed patterns similar to those in Japanese. These results are not contrary to the presumption that Mongolians and Japanese may have a common racial background.

Alleles↗

Sandwich-dot immunobinding assay (Sandwich-DIA), a new immunological method for the detection of diphtheria toxin.

In this paper we describe a new assay for diphtheria toxin in bacterial cultures, based on a sandwich-dot immunobinding method. This method uses horse polyclonal diphtheria antitoxin as the coating antibody and mouse monoclonal diphtheria antitoxin as the detecting antibody. The sensitivity of this new method is within a range of 5-10 ng toxin per ml in Corynebacterium diphtheriae cultures. We did not observe any false-positive reactions. As a solid phase we used nitrocellulose disks which bind protein more strongly than the commonly used plastic solid phase. Results with strong toxin producers of C. diphtheriae can be obtained as soon as 11 h after starting a bacterial test culture. To detect weak toxin producers, the cultivation period must be extended to about 18 h and the results can be obtained after about 24 h. The method presented in this paper is simpler, faster and more reliable than currently used methods.

Corynebacterium diphtheriae↗

Structural and mechanistic insights into the interaction between Rho and mammalian Dia.

Formins are involved in a variety of cellular processes that require the remodelling of the cytoskeleton. They contain formin homology domains FH1 and FH2, which initiate actin assembly. The Diaphanous-related formins form a subgroup that is characterized by an amino-terminal Rho GTPase-binding domain (GBD) and an FH3 domain, which bind somehow to the carboxy-terminal Diaphanous autoregulatory domain (DAD) to keep the protein in an inactive conformation. Upon binding of activated Rho proteins, the DAD is released and the ability of the formin to nucleate and elongate unbranched actin filaments is induced. Here we present the crystal structure of RhoC in complex with the regulatory N terminus of mammalian Diaphanous 1 (mDia1) containing the GBD/FH3 region, an all-helical structure with armadillo repeats. Rho uses its 'switch' regions for interacting with two subdomains of GBD/FH3. We show that the FH3 domain of mDia1 forms a stable dimer and we also identify the DAD-binding site. Although binding of Rho and DAD on the N-terminal fragment of mDia1 are mutually exclusive, their binding sites are only partially overlapping. On the basis of our results, we propose a structural model for the regulation of mDia1 by Rho and DAD.

Amino Acid Motifs↗

ROCK and Dia have opposing effects on adherens junctions downstream of Rho.

Adherens junctions (AJs) are crucial for maintaining the integrity of epithelial tissues and are often disrupted during tumour progression. Rho family proteins have been shown to regulate adherens junctions. We find that activation of the effector kinase ROCK and acto-myosin contraction disrupts AJs downstream of Rho. In contrast, signalling through the Rho effector Dia1 is required to ensure a dynamically stable interface between cells and the maintenance of adherens junction complexes. The ability of Dia1 to regulate the actin network is crucial for the localization of adherens junction components to the cell periphery.

Actins↗