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Development of a chemically defined liquid medium for growth of Legionella pneumophila.

A chemically defined liquid medium has been developed for the study of the physiology and antigen production of the Legionnaires disease bac … and ethylenediaminetetraacetic acid totally inhibited growth. Beta-mercaptoethanol, thioglycolate, dithiothreitol, and Tween 80 (0.05%) inhibited growth strongly or completely. Catalase activity was extremely weak or absent. Morphology varied, depending upon conditions and phases of growth. In general, filamentous forms became chains of cigar-shaped bacilli fra …

Международная многоцентральная оценка нового антидиотипического антидаратумумаба в целях устранения помех перед трансмиссией.

СПРАВОЧНАЯ ИНФОРМАЦИЯ И ЦЕЛИ: Даратумумаб, терапевтический человек anti-CD38 Моноклональные антитела улучшают множественные результаты миэломы, но препятствуют предварительному тестированию с помощью CD38 на реагентных красных кровяных клетках (RBC), потенциально маскируя клинически значимые аллоантитела. Цель этого исследования состояла в том, чтобы оценить эффективность нового антидиотипного антидаратумумабного реагента высокого уровня в деле нейтрализации интерференции даратумумаба, сохраняя при этом обнаружение аллоантиорганических свойств РБЦ и обеспечивая совместимость с обычными лаборат

Совпадение в аннотации оригинала
BACKGROUND AND OBJECTIVES: Daratumumab, a therapeutic human anti-CD38 monoclonal antibody, improves multiple myeloma outcomes but interferes … etection, cross-match resolution and workflow integration were assessed, alongside comparisons with dithiothreitol (DTT), papain, trypsin and DaraEx. RESULTS: Daratumumab interference was neutralized in 99.5% of samples, with 86.2% resolved using the 10% protocol. Detection of 190/197 RBC antibodies (96.4%) was preserved, including weak/low-titre antibodies. Ne …

[Enzyme Digestion LC-MS/MS Detection of Etomidate, Ketamine, Methamphe‑ tamine and Codeine in Hair].

OBJECTIVES: To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS)method based on enzymatic digestion for the rapid qualitative and quantitative detection of drugs in hair. METHODS: The enzymatic digestion parameters for etomidate, ketamine, methamphetamine, and codeine, including dithiothreitol (DTT) concentration, proteinase K concentration, incubation temperature and digestion time were optimized using controlled-variable experiments. After digestion, hair samples were filtered through a 0.22 μm membrane and separated on a C18 column with a gradient elution using 0.1% for…

Multiplex digital PCR for rapid quantitative detection and dynamic monitoring of respiratory pathogens in bronchoalveolar lavage fluid.

Lower respiratory tract infections (LRTIs) remain a leading cause of mortality worldwide. Rapid and comprehensive etiological diagnosis is critical yet challenging: existing approaches, from culture to multiplex PCR and mNGS, carry distinct limitations in one or more of turnaround time, sensitivity, cost, or quantitative resolution. We developed a 33-plex digital PCR (dPCR) assay for absolute quantification of bacteria, viruses, fungi, and atypical pathogens in bronchoalveolar lavage fluid (BALF). Systematic evaluation of five liquefaction protocols identified dithiothreitol (DTT) as the optim…

Personal exposure to particulate matter oxidative potential and airway inflammation: differences between asthmatic and non-asthmatic adults.

We aimed to determine whether exposure to particulate matter PM, measured as the oxidative potential (OP) of filters collected from 24 h personal samplers, is associated with increased airway inflammation in asthmatic and non-asthmatic volunteers. Forty-two adult asthmatic patients (25 women and 17 men) and 37 matched controls wore a personal sampler for a day collecting fine (PM2.5) and coarse (PM10-2.5) particles, and determining 24 h afterwards their fractional exhaled nitric oxide (FeNO). The PM-OP was determined by two methods: dithiothreitol (DTT) and ascorbic acid (AA) being OP levels d…

Improved recombinant expression of soluble cathepsin B and L in Escherichia coli.

Cysteine cathepsins such as cathepsin B and L play an important role in numerous diseases like acute pancreatitis or SARS-CoV-2 and therefor … elds for procathepsin B, and 37 ± 2 mg L-1 for procathepsin L, were achieved. After activation with dithiothreitol at slightly acidic pH, in vitro kinetic parameters of both cathepsins were determined using the commonly used synthetic substrates Arg-Arg-AMC or Phe-Arg-AMC. Moreover, to investigate the impact of the short C-terminal propeptide of procathepsin B, …

Pancreatic enzymes digest obstructive meconium from cystic fibrosis pig intestines.

INTRODUCTION: Meconium ileus (MI) is a life-threatening obstruction of the intestines affecting ∼15% of newborns with cystic fibrosis (CF). … , including normal saline, 7% hypertonic saline, and the reducing agents N-acetylcysteine (NAC) and dithiothreitol (DTT). We digested meconium at 37 °C with agitation, and measured meconium pigment release by spectrophotometry and residual meconium solids by filtration. RESULTS AND DISCUSSION: In CF pigs, meconium appeared as a solid pigmented mass obstructing …

Preparation of a Convertible Spacer Containing a Disulfide Group for Versatile Functionalization of Oligonucleotides.

The protocols described in this article provide details regarding the synthesis and characterization of a disulfide containing linker phosph … glycol) which shows concentration dependent release of the oligonucleotide upon treatment with 1,4-dithiothreitol, a reducing agent. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Preparation of disulfide linker phosphoramidite 3 Basic Protocol 2: Synthesis, functionalization, and characterization of DNA oligonucleot …

Effects of imidacloprid on viability and increase of reactive oxygen and nitrogen species in HepG2 cell line.

Imidacloprid (IMD) is a neonicotinoid insecticide used in large quantities worldwide in both veterinary and agronomic applications. Several … ed/oxidized glutathione ratio (GSH/GSSG), indicating oxidative stress. Furthermore, the antioxidant dithiothreitol, which reacts with ROS/RNS and acts as a thiol reducing agent, inhibited the cytotoxic effect of IMD. In addition, the metabolite IMD-olefin was more toxic than IMD. Our results indicate that IMD induces cytotoxicity in HepG2 cells and that this eff …

Hydrogen sulfide inhibited L-type calcium channels (CaV1.2) via up-regulation of the channel sulfhydration in vascular smooth muscle cells.

Hydrogen sulfide (H2S) exerts different effects on the cardiovascular system by modulating ion channels. The present study was to ascertain … ulfhydryl modifier diamide (DM) and significantly counteracted by the reductant sulfhydryl modifier dithiothreitol (DTT). DTT also partly reversed the NaHS-reduced [Ca2+]i in CaV1.2-HEK293 cells. Additionally, NaHS did not change CaV1.2 expression. Furthermore, NaHS increased phosphorylation of PKC and ERK in both a concentration- and a time-dependent manner in …

Associations between microvascular function and short-term exposure to traffic-related air pollution and particulate matter oxidative potential.

BACKGROUND: Short-term exposure to ambient air pollution has been associated with acute increases in cardiovascular hospitalization and mort … elemental carbon, and hopanes), and PM oxidative potential as quantified in two independent assays (dithiothreitol and in vitro macrophage ROS) in accumulation and ultrafine PM, and transition metals. CONCLUSIONS: Our findings suggest that short-term exposures to traffic-related air pollutants with high oxidative potential are major components contributing to m …

TRPA1 receptor stimulation by hydrogen peroxide is critical to trigger hyperalgesia and inflammation in a model of acute gout.

Acute gout attacks produce severe joint pain and inflammation associated with monosodium urate (MSU) crystals leading to oxidative stress pr … elicited responses were also reduced by the H2O2-detoxifying enzyme catalase and the reducing agent dithiothreitol. TRPA1 activation by MSU challenge-generated H2O2 mediates the entire inflammatory response in an acute gout attack rodent model, thus strengthening the role of the TRPA1 receptor and H2O2 production as potential targets for treatment of acute gout …

Sensory nerve terminal mitochondrial dysfunction activates airway sensory nerves via transient receptor potential (TRP) channels.

Mitochondrial dysfunction and subsequent oxidative stress has been reported for a variety of cell types in inflammatory diseases. Given the … ex III Qo site inhibitor) inhibited antimycin A-induced TRPA1 activation, as did the reducing agent dithiothreitol. Scavenging of both superoxide and hydrogen peroxide inhibited TRPA1 activation following mitochondrial modulation. In conclusion, we present evidence that acute mitochondrial dysfunction activates airway sensory nerves preferentially via TRPA1 thro …

Impaired macrophage phagocytosis in non-eosinophilic asthma.

BACKGROUND: Many patients with non-eosinophilic asthma have increased numbers of neutrophils in the airways. The explanation for this chroni … luding allergy skin tests, saline challenge and sputum induction. Sputum cells were dispersed using dithiothreitol and resuspended in culture medium. Efferocytosis of apoptotic bronchial epithelial cells by sputum-derived macrophages was determined using flow cytometry. RESULTS: There were no significant differences in efferocytosis between paired sputum and br …

Regulation of dipeptidyl peptidase 8 and 9 expression in activated lymphocytes and injured liver.

AIM: To investigate the expression of dipeptidyl peptidase (DPP) 8 and DPP9 in lymphocytes and various models of liver fibrosis. METHODS: DPP8 and DPP9 expression were measured in mouse splenic CD4⁺ T-cells, CD8⁺ T-cells and B-cells (B220⁺), human lymphoma cell lines and mouse splenocytes stimulated with pokeweed mitogen (PWM) or lipopolysaccharide (LPS), and in dithiothreitol (DTT) and mitomycin-C treated Raji cells. DPP8 and DPP9 expression were measured in epidermal growth factor (EGF) treated Huh7 hepatoma cells, in fibrotic liver samples from mice treated with carbon tetrachloride (CCl₄)…

D-Penicillamine targets metastatic melanoma cells with induction of the unfolded protein response (UPR) and Noxa (PMAIP1)-dependent mitochondrial apoptosis.

D-Penicillamine (3,3-dimethyl-D-cysteine; DP) is an FDA-approved redox-active D-cysteine-derivative with antioxidant, disulfide-reducing, an … ermal melanocytes, an effect not observed with the thiol-antioxidants N-acetyl-L-cysteine (NAC) and dithiothreitol. Focused gene expression array analysis followed by immunoblot detection revealed that DP rapidly activates the cytotoxic unfolded protein response (UPR; involving phospho-PERK, phospho-eIF2α, Grp78, CHOP, and Hsp70) and the mitochondrial pathway of …

Nitric oxide selectively depletes macrophages in atherosclerotic plaques via induction of endoplasmic reticulum stress.

BACKGROUND AND PURPOSE: Macrophages in atherosclerotic plaques have a tremendous impact on atherogenesis and plaque destabilization. We prev … s well as with the well-known endoplasmic reticulum (ER) stress inducers thapsigargin, tunicamycin, dithiothreitol or brefeldin A. Cell viability was analysed by Neutral Red viability assays. Cleavage of caspase-3, DNA fragmentation and ultrastructural changes were examined to characterize the type of macrophage death. Induction of ER stress was evaluated by mea …

Tyrosine phosphorylation generates multiple isoforms of the mitochondrial capsule protein, phospholipid hydroperoxide glutathione peroxidase (PHGPx), during hamster sperm capacitation.

Sperm capacitation is a maturation process, occurring in the female reproductive tract, that produces fertilization-competent spermatozoa. P … - 99,000), a 19-kDa polypeptide was the only one that can be solubilized completely by Triton X-100-dithiothreitol (DTT). The 19-kDa polypeptide was purified by anion-exchange chromatography and by immunoaffinity chromatography. Proteomic identification of the 19-kDa polypeptide by nano-electrospray tandem mass spectrometry yielded six peptides that matched the …

Local production of Aspergillus fumigatus specific immunoglobulin E in nasal polyps.

BACKGROUND: Aspergillus spp. play a significant role in the etiology of immunoglobulin (Ig)E mediated allergic fungal sinusitis (AFS). It is unclear whether Aspergillus spp. are also involved in nasal polyps without the characteristic clinical features of AFS. OBJECTIVES: The frequency of Aspergillus spp. and Aspergillus-specific IgE in nasal lavages and serum of patients with severe nasal polyps (n = 33) without clinical features of AFS should be investigated. STUDY DESIGN: Prospective study. METHODS: An aliquot of nasal lavage fluid was treated with dithiothreitol and examined for Aspergi…

Presentation of nitric oxide regulates monocyte survival through effects on caspase-9 and caspase-3 activation.

In the absence of survival factors, blood monocytes undergo spontaneous apoptosis, which involves the activation of caspase-3. Although nitr … ne directly blocked the activity of recombinant caspase-3, which was reversed by the reducing agent dithiothreitol, whereas PAPA or DEA NONOate did not block the enzymatic activity of caspase-3. These data support the hypothesis that nitrosylation of protein thiol residues by nitric oxide is critical for promoting the survival of human monocytes.