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[Studies of occurrence of antigen Dia from the Diego blood-group system in the Polish population].

In 9661 blood donors from seven regions of Poland studies were carried out for detection of the Dia antigen in erythrocytes. This antigen was found in 45 (0.46%) subjects not consanguinous. The prevalence of Dia in various regions of the country varied from 0.25 to 0.91%. Family studies in 23 cases with 94 subjects participating demonstrated inheritance of the antigen. It was tried to explain the presence of the Dia in the Polish population, since in the pertinent literature it is regarded as a characteristic trait of Mongoloid peoples.

Blood Donors↗

Development and preliminary evaluation of an IgM dot-immunobinding assay (IgM-DIA) for rapid serodiagnosis of scrub typhus infection.

An IgM dot-immunobinding assay (IgM-DIA) was developed for the diagnosis of scrub typhus infection. The whole cell antigens of Karp, Kato and Gilliam strains of Rickettsia tsutsugamushi were immobilized onto nitrocellulose paper and reacted with patients sera. The presence of IgM R. tsutsugamushi specific antibody in the patient sera could be detected by the observation of a visible brown dot on the nitrocellulose paper. The IgM-DIA has a sensitivity of 90.4% and specificity of 81.4% as compared to the indirect immunoperoxidase test. The IgM-DIA is rapid, simple, cost-effective, does not require microscope or incubator. It is recommended as a rapid screening test for the diagnosis of scrub typhus infection in the field or rural area within the hyperendemic region.

Antibodies, Bacterial↗

The Desmoteplase in Acute Ischemic Stroke Trial (DIAS): a phase II MRI-based 9-hour window acute stroke thrombolysis trial with intravenous desmoteplase.

BACKGROUND AND PURPOSE: Most acute ischemic stroke patients arrive after the 3-hour time window for recombinant tissue plasminogen activator (rtPA) administration. The Desmoteplase In Acute Ischemic Stroke trial (DIAS) was a dose-finding randomized trial designed to evaluate the safety and efficacy of intravenous desmoteplase, a highly fibrin-specific and nonneurotoxic thrombolytic agent, administered within 3 to 9 hours of ischemic stroke onset in patients with perfusion/diffusion mismatch on MRI. METHODS: DIAS was a placebo-controlled, double-blind, randomized, dose-finding phase II trial. Patients with National Institute of Health Stroke Scale (NIHSS) scores of 4 to 20 and MRI evidence of perfusion/diffusion mismatch were eligible. Of 104 patients, the first 47 (referred to as Part 1) were randomized to fixed doses of desmoteplase (25 mg, 37.5 mg, or 50 mg) or placebo. Because of an excessive rate of symptomatic intracranial hemorrhage (sICH), lower weight-adjusted doses escalating through 62.5 microg/kg, 90 microg/kg, and 125 microg/kg were subsequently investigated in 57 patients (referred to as Part 2). The safety endpoint was the rate of sICH. Efficacy endpoints were the rate of reperfusion on MRI after 4 to 8 hours and clinical outcome as assessed by NIHSS, modified Rankin scale, and Barthel Index at 90 days. RESULTS: Part 1 was terminated prematurely because of high rates of sICH with desmoteplase (26.7%). In Part 2, the sICH rate was 2.2%. No sICH occurred with placebo in either part. Reperfusion rates up to 71.4% (P=0.0012) were observed with desmoteplase (125 microg/kg) compared with 19.2% with placebo. Favorable 90-day clinical outcome was found in 22.2% of placebo-treated patients and between 13.3% (62.5 microg/kg; P=0.757) and 60.0% (125 microg/kg; P=0.0090) of desmoteplase-treated patients. Early reperfusion correlated favorably with clinical outcome (P=0.0028). Favorable outcome occurred in 52.5% of patients experiencing reperfusion versus 24.6% of patients without reperfusion. CONCLUSIONS: Intravenous desmoteplase administered 3 to 9 hours after acute ischemic stroke in patients selected with perfusion/diffusion mismatch is associated with a higher rate of reperfusion and better clinical outcome compared with placebo. The sICH rate with desmoteplase was low, using doses up to 125 microg/kg.

Acute Disease↗

Is the Mnr locus of Triticeae species the same as the Ndh and Dia loci?

The menadione reductase (MNR), the nicotinamide adenine dinucleotide dehydrogenase (NDH) and diaphorase (DIA) isozymes were studied in the allohexaploid Triticum aestivum cv "Chinese Spring" and in five diploid Triticeae species. The Mnr1, Ndh3and Dia1 loci were located on the chromosome arms 3AL, 3BL and 3DL of T. aestivum, respectively. These loci were also located on the 3H chromosome of Hordeum vulgare cv "Betzes", the 3L chromosome of Aegilops longissima and the 6RL chromosome arm of Secale cereale cv "Imperial". The chromosomal location results together with the segregation studies support a tetrameric behaviour of the MNR1, NDH3 and DIA1 isozymes. The Ndh1 and Dia3 loci were located on homoeologous group 4 showing a monomeric behaviour. The chromosomal locations and linkage data of the Mnr, Ndh and Dia loci suggest that Mnr1=Ndh3=Dia1; Ndh1=Dia3 and Ndh2=Dia2.

Journal Article↗

Urea removal by neonatal hemo(dia)filters in hyperazotemic rabbits.

OBJECTIVE: Comparison of three neonatal hemo(dia)filters (FH22, Gambro; Minifilter Plus, Amicon; Miniflow 10, Hospal) for removal of urea by venovenous hemofiltration (HF) and venovenous hemodiafiltration (HDF). DESIGN: Filters were successively used for HF with two different blood flows (Qb = 8.3 and 16.6 ml/ min) and for HDF with the two different blood flows and four dialysate flows (Qd = 0.5, 1.0, 2.0, and 3.0 l/h). SUBJECTS: 21 anesthetized adult New Zealand White rabbits infused with urea. MAIN RESULTS: Urea clearance was significantly increased by HDF compared to HF regardless of blood flow, dialysate flow, and the hemo (dia)filter type except in the FH22 group, when blood flow was high and dialysate flow was 0.5 or 1.0 l/h. The FH22 filter allowed the best urea clearance during HF at high blood flow. During the HDF procedures, the Miniflow 10 allowed the highest urea clearance regardless of blood flow and dialysate flow.

Analysis of Variance↗

Automatic quantitation of HIV-1 mediated cell-to-cell fusion with a digital image analysis system (DIAS): application for rapid screening of HIV-1 fusion inhibitors.

Human immunodeficiency viruses type 1 (HIV-1) mediated cell-to-cell fusion plays an important role in HIV-1 spread from infected cells to uninfected cells and in HIV-1 cytopathogenesis. In the present study, we developed a convenient cell fusion assay using a computer-controlled digital image analysis system (DIAS) for automatic quantitation. Compared with a manual quantitative method, DIAS automatic method is less laborious, and more rapid. Furthermore, it is more objective and less dependent on the researchers' experience. This method has great potential to be developed further as a high-throughput screening assay for rapid screening of HIV-1 fusion inhibitors, for evaluating the activity of HIV-1 entry inhibitors and for studying the mechanism of action of anti-HIV-1 agents.

Cell Fusion↗

From Peaks to Power: Systematic Evaluation of Chromatographic Sampling Reveals Determinants of Quantification and Biological Discovery in DIA Proteomics.

Modern DIA proteomics increasingly emphasizes throughput and depth for large-cohort studies, but methods are often optimized using proxy metrics that can mask losses in quantifiable signal and statistical power. Here, we evaluate how data points per peak and other chromatographic features jointly contribute to quantification and downstream biological discovery. Using a matrix-matched calibration curve dataset, we checked how the number of data points per peak (DPPP) affects the limits of detection and quantification (LOD/LOQ). Reduced DPPP minimally affected LOD but substantially degraded LOQ. Feature modeling and nonparametric association analyses identified precursor peak area as the strongest feature-level predictor of LOQ, whereas DPPP showed weaker and context-dependent effects. Simulations of chromatographic peak integration recapitulated these trends, showing that increased sampling primarily improves integration precision, while quantitative accuracy is strongly governed by peak height and peak shape. Finally, when comparing 20 cancer vs 20 control plasma samples processed with Seer Proteograph, the decrease in DPPP led to a loss of statistical significance for proteins with low-abundance precursors. These findings argue that DIA optimization should prioritize LOQ and statistical power metrics─not identifications alone─by balancing sampling density with chromatographic peak height and quality to maximize useful biological signal.

Proteomics↗

DIA proteomics of FFPE renal biopsies reveals two molecular subtypes of lupus nephritis and identifies APOL1 as candidate biomarker for stratification.

INTRODUCTION: Lupus nephritis (LN) exhibits substantial clinical and pathological heterogeneity. We aimed to define proteomics-based molecular subtypes of LN and identify candidate biomarkers for subtype discrimination. METHODS: We analysed formalin-fixed paraffin-embedded (FFPE) renal biopsy specimens from 292 patients with biopsy-proven LN from four tertiary hospitals using data-independent acquisition (DIA)-liquid chromatography-tandem mass spectrometry (LC-MS/MS) proteomics. Molecular subtypes were identified by non-negative matrix factorisation. Differential proteins, functional enrichment, immune pathway activity, protein-protein interaction networks and subtype-associated clinical/pathological features were evaluated. Extreme Gradient Boosting (XGBoost) with SHapley Additive exPlanations (SHAP) and Least Absolute Shrinkage and Selection Operator (LASSO) logistic regression were used to identify key subtype-related features and derive a protein panel distinguishing proliferative (class III/IV) from membranous (class V) LN. RESULTS: Two stable molecular subtypes were identified, with 1002 differential proteins between them. Subtype_2 was enriched for interferon-related innate immunity, complement activation, phagocytosis-endocytosis-lysosome pathways and ribosome biogenesis/RNA metabolism, whereas Subtype_1 was characterised by keratinisation and epithelial structural remodelling. Subtype_2 was associated with higher serum creatinine, lower estimated glomerular filtration rate and higher chronicity index. APOL1 showed discriminatory value between subtypes, and serum ELISA demonstrated a consistent pattern with FFPE proteomic findings. A five-protein LASSO panel achieved an area under the curve of approximately 0.76 for distinguishing class III/IV from class V LN. CONCLUSION: DIA-based proteomic profiling of FFPE renal biopsies identifies biologically and clinically relevant LN molecular subtypes and may support tissue-informed classification and risk stratification.

Humans↗

Application of 2D and 3D DIAS to motion analysis of live cells in transmission and confocal microscopy imaging.

The chemotactic signal in Dictyostelium is a cAMP wave that is relayed over relatively large distances through a cell population during aggregation. Cells exhibit unique behaviors in response to the different spatial, temporal, and concentration components of the cAMP wave, suggesting that distinct signal transduction pathways are evoked in each of the various phases of the wave. For this reason, we designed a set of experimental protocols to test responses of normal and mutant Dictyostelium amoebae to the different components of a wave of chemoattractant. We then used computer-assisted two- (2D) and three-dimensional (3D) technologies (2D and 3D Dynamic Image Analysis System [DIAS]) for analysis of cells in the absence of a chemotactic signal (basic motile behavior) and in response to the temporal, spatial, and concentration components of the wave. As a result, we have elucidated parallel and independent pathways activated by specific phases of the cAMP wave. Likewise, human polymorphonuclear neutrophils (PMNs) respond to experimentally applied waves of the chemotactic peptide fMLP, and also exhibit discrete behavioral responses to the different phases. Using Dictyostelium as a paradigm, we applied our protocols to normal and diseased human PMNs and precisely defined a chemotactic defect. In this chapter, we describe methods for quantifying behaviors in Dictyostelium amoebae, PMNs, and other amoeboid cells using 2D and 3D DIAS. These methods are useful in the reconstruction and motion analysis of most migrating cells with transmitted and/or confocal microscopy.

Animals↗

A delayed hemolytic transfusion reaction (DHTR) with multiple alloantibodies (Anti-E, Jka, Dia, Fyb, and S) induced by E-antigen-negative, crossmatch-compatible blood.

We describe the case of a 44-year-old woman with a delayed hemolytic transfusion reaction (DHTR). She had a history of two pregnancies and a blood transfusion, the details of which were unknown. At the time of her first vascular surgery on November 15, 1989, she received 1200 ml of crossmatch-compatible concentrated red blood cells (CRC). Before the first operation, screening for anti-RBC antibodies (Ab) was negative. At the time of the second admission on Feburary 15, 1996, anti-E Abs were detected by indirect antiglobulin test. She received 560 ml of E-antigen-negative, crossmatch-compatible, CRC for treatment of anemia on March 1 and 2, 1996. After this transfusion, total bilirubin (1.6 mg/dl) and lactate dehydrogenase (1355 IU/ml) were elevated on March 12, 1996. She had no evidence of clinical hemolysis. We suspected DHTR from these data, and therefore screened for anti-RBC Abs. Anti-E, Jka, Dia, Fyb, and S Abs were detected in blood samples obtained from the patient on March 12, 1996. Anti-E, Jka Dia, and S Abs were present more than 1 month and anti-Fyb Ab was disappeared at 18 days after transfusion.

Adult↗

Case report. An immediate hemolytic transfusion reaction apparently caused by anti-Dia.

The Diego blood group system has had its primary applications in population genetics and anthropology, although it can also give rise to clinical problems. Anti-Dia has ofter been reported to cause hemolytic disease of the newborn. The patient presented in the report experienced an immediate hemolytic transfusion reaction apparently due to anti-Dia. We believe it to be the only such case reported since the Diego system was first discovered in 1956.

Adult↗

The first example of anti-Dia antibody in Poland.

A strong incomplete and complement-fixing anti-Dia within Diego blood group system was found in the serum of a Polish mother who gave birth to a newborn suffering from a severe haemolytic disease. In family studies of three generations Dia antigen in three persons was detected and its hereditary character was confirmed. The whole family is of Polish origin.

Blood Group Antigens↗

Band 3 Memphis variant II. Altered stilbene disulfonate binding and the Diego (Dia) blood group antigen are associated with the human erythrocyte band 3 mutation Pro854-->Leu.

Band 3 Memphis is a commonly occurring polymorphic form of the human red cell anion transporter (band 3, AE1). Band 3 Memphis migrates more slowly on an SDS-polyacrylamide gel than normal band 3 and results from a point mutation Lys56-->Glu. Two types of band 3 Memphis, variants I and II, can be distinguished by their susceptibility to covalent labeling with H2DIDS (4,4'-diisothiocyanato-2,2'-dihydrostilbene disulfonate). Memphis variant II is more readily labeled than Memphis variant I or normal band 3. The Memphis variant II is also associated with the presence of the Diego (Dia) blood group antigen on the red cells. We have shown that Memphis variant II carries the polymorphism Pro854-->Leu, as well as Lys56-->Glu. The blood group antigen (Dia) present at the surface of Memphis variant II type red cells suggests the mutation Pro854-->Leu causes a change in the structure of an extracellular loop of Memphis variant II band 3. We discuss possible ways in which the mutation Pro854-->Leu affects the reactivity of Lys539 to covalent reaction with H2DIDS.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Molecular mechanisms underlying umami taste perception: A DIA-based proteomic analysis of Agrocybe aegerita peptides.

The mechanisms underlying the modulation of the salivary perception of umami peptides remain poorly understood. Herein, three umami peptides (DDL, DEL, and ENG) obtained from Agrocybe aegerita were used to investigate the regulatory role of saliva in umami taste perception via a combined approach involving sensory evaluation and proteomics analysis based on 4D-DIA technology. The results revealed that umami intensity peaked at 10&#xa0;s after ingestion and was accompanied by a significant increase in saliva secretion (p&#xa0;<&#xa0;0.05). Further proteomics analysis revealed that lactotransferrin and proline-rich proteins are closely associated with the sensory perception of umami peptides. Differentially expressed proteins were mainly enriched in pathways related to saliva secretion and proteasome function. This study provides new insights from the perspectives of salivary proteomics and dynamic salivary secretion, contributing to a deeper understanding of the mechanisms by which saliva regulates umami perception.

Humans↗

Temporal DIA-MS proteomics reveals coordinated metabolic reprogramming associated with oil accumulation in oil palm mesocarp.

Oil palm (Elaeis guineensis Jacq.) is the most productive oil-bearing crop globally, yet the molecular basis of mesocarp development and lipid accumulation remains poorly understood. Ultra-deep data-independent acquisition mass spectrometry (DIA-MS) was applied to characterize proteome dynamics in two contrasting genotypes, seedless (KS) and thin-shelled (TS), across five developmental stages (P1-P5) spanning fruit development to mature oil accumulation. Phenotypic analysis revealed higher mesocarp proportion and oil content in KS during late maturation. A total of 137,615 peptides corresponding to 12,163 protein groups were identified, providing a temporal proteomic landscape of mesocarp development. Multivariate analysis indicated that developmental progression was the primary contributor to proteomic variation, whereas genotype-associated differences increased during lipid accumulation. Differentially abundant proteins were mainly associated with carbohydrate metabolism, photosynthesis, proteolysis, antioxidant responses, and lipid biosynthesis. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and KOG analyses suggested extensive remodeling of metabolic networks, including developmental changes in photosynthesis-associated proteins and increased representation of lipid-associated pathways during maturation. Weighted protein co-expression network analysis identified 17 modules associated with developmental progression and lipid accumulation, highlighting candidate proteins involved in carbon metabolism, energy production, and cellular protection. Genes encoding selected hub protein candidates were further examined by RT-qPCR. Biochemical analyses supported these proteomic patterns, showing increased acetyl-CoA availability, enhanced antioxidant enzyme activities (SOD, CAT, APX, and GR), improved GSH/GSSG balance, and reduced oxidative damage in KS. Together, these findings provide a temporal proteomic and biochemical framework for understanding genotype-associated differences in oil accumulation and identify candidate metabolic networks for functional studies.

Carbon metabolism↗

Time- and Dose-Resolved DIA-PASEF Proteomics Maps the Transition from Adaptive Stress to Apoptotic Collapse in Melittin-Treated MDA-MB-231 Cells.

Melittin, the cytolytic peptide of honeybee venom, exhibits potent anticancer activity in triple-negative breast cancer (TNBC), yet the molecular programs underlying its cytotoxic effects remain incompletely defined. To address this gap, MDA-MB-231 TNBC cells were exposed to melittin at half-maximal inhibitory concentration(half IC50) and IC50 across early(0.5, 1, and 2 h), mid(3, 4 h), and late (12, 24 h) time windows. Proteomic profiling was performed using label-free data-independent acquisition(DIA) parallel accumulation-serial fragmentation(PASEF). Approximately 5800 proteins were quantified, revealing distinct dose-dependent stress responses. An integrative exploratory framework combining time-resolved log2 fold-change trajectories, area-under-the-curve(AUC) based temporal prioritization, and independent heatmap visualization identified proteins associated with melittin-induced stress remodeling. Half IC50 exposure showed a transient stress-adaptive signature characterized by chromatin remodeling(HMGN2, H2AZ1), structural and RNA-associated buffering(LRRC7), and indirect mitochondrial quality-control signaling(CPAMD8, SPATA4), which progressively weakened over time. In contrast, IC50 treatment induced rapid chromatin remodeling dominated by histone H1 variants(H1.4, H1.2), early RNA instability(LRRC7), and late-stage cytoskeletal disassembly marked by MICAL3 induction, consistent with progression toward apoptosis. These trajectories paralleled dose-dependent apoptotic phenotypes. Overall, data suggest that melittin elicits dose- and time-dependent proteomic stress responses in TNBC cells and identify candidate trajectory-associated proteins and pathways linked to adaptive stress remodeling or progression toward cytotoxic collapse.

Melitten↗

Dia-metaphyseal distal tibial fractures--treatment with a shortened intramedullary nail: a review of 15 cases.

Fifteen patients were reviewed retrospectively with distal tibial fractures treated with a reamed intramedullary nail which had approximately 1 cm removed just distal to the lowermost locking screw. There were 15 patients (nine males and six females). All patients had returned to normal activities of daily living. Eleven patients could perform all leisure activities with no symptoms and three had only minor discomfort, which did not preclude sport. All fractures united, 12 uneventfully and three after a secondary surgical procedure. Three patients had malalignment defined as varus-valgus angulation or recurvatum of 5 degrees or greater. Whilst technically challenging, intramedullary nailing for dia-metaphyseal distal tibial fractures represents a safe and reliable method for managing these injuries. This represents the first report in the English Language literature specifically examining treatment with a shortened tibial nail.

Adult↗