Der BCL-2 Inhibitor ABT-199/Venetoclax transaktiviert NOXA und wirkt mit Proteasom-Inhibition bei der Zelltod-Induktion synergistisch

The development of neoplastic transformations is characterized by persistent proliferation as well as deregulation of the apoptosis signalling pathway. Reduced apoptosis is often caused by overexpression of anti-apoptotic proteins of the BCL 2 protein family. Consequently, these are therapeutic targets, which can be inhibited by specific BCL 2 inhibitors. The specific BCL 2 inhibitor ABT 199 is effective in Multiple Myeloma (MM). Also, the proteasome inhibitor BTZ is an effective and established anti-cancer therapeutic agent approved for therapy of MM and specifically for mantle cell lymphoma (MCL). We recently published that the specific BCL 2 inhibitor ABT 199 in combination with bortezomib (BTZ) is effective in inducing cell death in solid tumours, especially in soft-tissue sarcomas (STS). Further investigation of the identified synergistic apoptosis induction by ABT 199 and BTZ revealed that the mechanism of action is not BTZ-specific but rather mediated by proteasome inhibition in general. The verified key molecules in the ABT 199&PI (proteasome inhibitor)-mediated cell death induction are the effector protein BAX and the BH3-only protein NOXA. Furthermore, the present study shows for the first time that ABT 199 markedly increases PMAIP1/NOXA expression through transcriptional induction. The ABT 199-mediated transcriptional induction of NOXA is predominantly TP53-independent. Analyses of mRNA and protein expression show that ABT-199 induces PMAIP1/NOXA expression through activation of the integrated stress response (ISR). In summary, ABT 199 promotes apoptosis in a twofold manner: i) ABT-199 blocks anti-apoptotic BCL 2 and ii) ABT 199 transactivates NOXA which inhibits anti-apoptotic MCL-1. The identified synergism comes into play when NOXA degradation is additionally inhibited by proteasome inhibition. Since the here revealed and formerly unidentified ABT 199-mediated activation of the ISR and transcriptional induction of NOXA is cell-type unspecific, this mechanism could find application not only in hematopoietic diseases, but also be effective in solid tumour entities

Detection and prevention of the dispersal of the seeds of Chilean needle grass (Nassella neesiana Trin. & Rupr. (Barkworth)) in hay bales

Invasive and noxious weeds are well known as a pervasive problem, imposing significant economic burdens on all areas of agriculture, and thus their control and eradication are of central concern to the industry. Whilst there are multiple possible pathways of weed dispersal in this industry, of particular interest to this discussion is how the unintended dispersal of Chilean needle grass (Nassella neesiana) weed seeds, within emergency fodder, may be prevented. Periodic drought is a feature of the Australian climate, which, together with natural disasters of flood and bushfire, necessitates the urgent provision of emergency fodder for maintenance of livestock production. However, previous investigations into weed seed dispersal by this fodder type have revealed a significant risk for the dispersal of a number of noxious weeds well beyond their current range. Because preventive work, relating to detection and eradication of noxious species, is at an early stage of development, the development of faster, more reliable, and non-destructive sampling methods for seed species has become essential. In addition to prevention of weed seed dispersal in fodder, the use of appropriate methods of controlling troublesome pasture weeds in situ is of relevance for addressing established infestations. Whilst silage preservation is one such typical method that can reduce weed seed input into the soil seed bank, there is currently little data on the absolute effectiveness of this method for control of N. neesiana. To investigate whether rapid detection of seeds in bales is possible and how the numbers of seeds in a pasture are linked to the numbers of seeds that may be incorporated into hay bales made from an infested pasture, an investigation of a rapid, non-destructive technique to detect seeds in bales, together with an examination of a pasture to estimate seed density and dispersal prior to baling, were carried out. Additionally, since pastures infested with this weed are of no use for producing hay and little use for grazing, a third experiment was devised, which was an investigation of silaging for preventing inadvertent seed input into pasture soil seed banks. This thesis has shown that this approach of non-destructive core sampling of bales to detect the seeds of N. neesiana, and possibly others, which can be visually recognised, shows promise for those involved in weed control. Similarly, the simultaneous demonstration that silage fermentation is an efficient viable method for both eliminating viability of mature (germinable) seeds of this species while also reducing the probability of injury to livestock from these seeds, confirms that reducing the impact of this weed in agro-ecological systems and on livestock during emergency situations is possible.Doctor of Philosoph

Amaranthus retroflexus L (redroot pigweed) : effects of elevated CO2 and soil moisture on growth and biomass and the effect of radiant heat on seed germination

Amaranthus retroflexus L. (Amaranthaceae), Redroot pigweed, is native to North America, but has become a weed of agriculture worldwide. Previous research into competition with food crops found it significantly reduces yields. Additionally, taxonomy, biomass allocation, physiological responses to light intensity, water stress, elevated CO2, and herbicide resistance have been inves-tigated. To extend other research findings, we investigated growth and biomass yield in response to (i) soil moisture stress, and (ii) drought and elevated CO2. Additionally, we investigated seed germination rates following exposure to three elevated temperatures for two different time periods. Overall, moisture stress reduced plant height, stem diameter, and number of leaves. Elevated CO2 (700 ppm) appeared to reduce negative impacts of drought on biomass productivity. Heating seeds at 120◦C and above for either 180 or 300 s significantly reduced germination rate. These results inform an understanding of potential responses of A. retroflexus to future climate change and will be used to predict future occurrence of this weed. The finding that exposing seeds to high temperatures retards germination suggests fire could be used to prevent seed germination from soil seed banks, particularly in no-till situations, and therefore may be used to address infestations or prevent further spread of this weed. © 2021 by the authors. Licensee MDPI, Basel, Switzerland. **Please note that there are multiple authors for this article therefore only the name of the first 5 including Federation University Australia affiliates “Sandra Weller, Singarayer Florentine, Amali Welgama, Aakansha Chadha, Chrisopher Turville" are provided in this record*

DNA Polymerase η Is Involved in Hypermutation Occurring during Immunoglobulin Class Switch Recombination

Base substitutions, deletions, and duplications are observed at the immunoglobulin locus in DNA sequences involved in class switch recombination (CSR). These mutations are dependent upon activation-induced cytidine deaminase (AID) and present all the characteristics of the ones observed during V gene somatic hypermutation, implying that they could be generated by the same mutational complex. It has been proposed, based on the V gene mutation pattern of patients with the cancer-prone xeroderma pigmentosum variant (XP-V) syndrome who are deficient in DNA polymerase η (pol η), that this enzyme could be responsible for a large part of the mutations occurring on A/T bases. Here we show, by analyzing switched memory B cells from two XP-V patients, that pol η is also an A/T mutator during CSR, in both the switch region of tandem repeats as well as upstream of it, thus suggesting that the same error-prone translesional polymerases are involved, together with AID, in both processes

Somatic diversification in the absence of antigen-driven responses is the hallmark of the IgM+IgD+CD27+ B cell repertoire in infants

T cell–dependent immune responses develop soon after birth, whereas it takes 2 yr for humans to develop T cell–independent responses. We used this dissociation to analyze the repertoire diversification of IgM+IgD+CD27+ B cells (also known as “IgM memory” B cells), comparing these cells with switched B cells in children <2 yr of age, with the aim of determining whether these two subsets are developmentally related. We show that the repertoire of IgM+IgD+CD27+ B cells in the spleen and blood displays no sign of antigen-driven activation and expansion on H-CDR3 spectratyping, despite the many antigenic challenges provided by childhood vaccinations. This repertoire differed markedly from those of switched B cells and splenic germinal center B cells, even at the early stage of differentiation associated with μ heavy chain expression. These data provide evidence for the developmental diversification of IgM+IgD+CD27+ B cells, at least in very young children, outside of T cell–dependent and –independent immune responses

Prevalence and factors associated with sleep disturbance in adult patients with psoriasis

Background: Sleep, which is crucial for restoring of physiological functions and health, is reportedly impaired in psoriasis. The role of different potential sleep confounding factors, including detailed pruritus characteristics, and the complex interplay between psychological variables (anxiety and depression), pruritus and sleep disturbance in psoriasis remain insufficiently investigated. Objectives: To investigate sleep characteristics and to identify clinical, demographic and psychological factors associated with sleep disturbance in psoriasis. Methods: This cross-sectional study included 334 psoriasis patients (response rate 86%) and 126 control subjects (response rate 82%). Measures included sleep quality [Pittsburgh Sleep Quality Index (PSQI)], psoriasis severity, pruritus characteristics, including average pruritus intensity [visual analogue scale (VAS)], severity of comorbidities, anxiety and depression (Hospital Anxiety and Depression Scale - HADS) and quality of life (Dermatology Life Quality Index - DLQI, and Short Form 12 - SF12). Results: Fifty-nine per cent of patients and 34% of control subjects (P 5). Patients slept 1 h less than control subjects (median 6 vs. 7 h, P < 0.001). Patients without pruritus had less impaired sleep (global PSQI) than patients with strong (P < 0.001) and very strong pruritus (P < 0.001). Anxiety (HADS-A) and depression (HADS-D) levels were the strongest predictors of sleep impairment, followed by pruritus exacerbation at night, age, female sex, pruritus exacerbation in the morning, average pruritus intensity (VAS), diagnosed depression and gastroesophageal reflux disease, altogether explaining 32%-37% of the variance in global sleep quality. Both anxiety (HADS-A) and depression (HADS-D) were significant mediators explaining the association between pruritus intensity (VAS) and sleep impairment in 42% and 37% respectively. Conclusions: Sleep disturbance in patients with psoriasis is highly prevalent. Patients with psoriasis should be assessed for sleep impairment, pruritus, anxiety and depression. Reduction in pruritus should be considered as an important therapeutic goal, along with therapies aimed at reducing anxiety and depression

Catalysis of Strand Exchange by the HSV-1 UL12 and ICP8 Proteins: Potent ICP8 Recombinase Activity is Revealed upon Resection of dsDNA Substrate by Nuclease

The replication of herpes simplex virus type 1 (HSV-1) is associated with a high degree of homologous recombination, which is likely to be mediated, in part, by HSV-1-encoded proteins. We have previously shown that the HSV-1 encoded ICP8 protein and alkaline nuclease UL12 are capable of catalyzing an in vitro strand-exchange reaction. Here, we show, by electron microscopy, that the products of the strand exchange reaction between linear double-stranded DNA and circular single-stranded DNA consist of the expected joint molecule forms: sigma, alpha, and gapped circles. Other exonucleases, such as lambda Red α, which, like UL12, digests 5′-3′, as well as Escherichia coli exonuclease III (ExoIII), which digests 3′-5′, could substitute for UL12 in the strand exchange reaction by providing a resected DNA end. ICP8 generated the same intermediates and strand exchange products when the double-stranded DNA substrate was preresected by any of the nucleases. Using substrates with large regions of non-homology we found that pairing by ICP8 could be initiated from the middle of a DNA molecule and did not require a homologous end. In this reaction, the resection of a DNA end by the nuclease is required to reveal homologous sequences capable of being paired by ICP8. This study further illustrates the complexity of the multi-functional ICP8 protein

Adsorption of 2,2 '-dithiodipyridine as a tool for the assembly of silver nanoparticles

Silver nanostructured thin films stabilized by 2,2’-dithiodipyridine (2dtpy) were prepared. The Ag nanoparticles were obtained by treating the complex [Ag(2dtpy)]NO3 with NaBH4 in a methanol–toluene mixture. The films were transferred to borosilicate glass slips by a dip-coating method and were found to consist of Ag nanoparticles possibly linked via 2dtpy molecules. Surface-enhanced Raman scattering (SERS) studies have offered the possibility of investigating the adsorption modes of 2dtpy at the Ag nanoparticle surfaces in the fil

A Gel-Based Dual Antibody Capture and Detection Method for Assaying of Extracellular Cytokine Secretion: EliCell

A distinguishing feature of eosinophils is their ability to rapidly release preformed cytokines from intracellular pools. Cytokines are delivered to the cell surface from granule stores by transport vesicles and are released in small packets at discrete locations along the cell surface through a process termed “piecemeal” degranulation. The study of this process has been hindered by lack of an assay sensitive enough to register minute protein concentrations and the inability to visualize morphology of cytokine secreting cells. These hindrances have necessitated our development of the EliCell assay, an agarose-based dual cytokine capture and detection system through which cytokine secretion and cellular morphology may be analyzed in concert. Cells are embedded within capture antibody-containing agarose and stimulated under conditions of interest. Extracellularly released cytokine is captured within the matrix at the point of release from the cell and can be labeled with a fluorochrome-conjugated antibody. Cytokine release and cellular morphology are visualized in parallel by phase contrast and fluorescence microscopy, respectively

Response of Chloris truncata to moisture stress, elevated carbon dioxide and herbicide application

Herbicide resistance has been observed in Chloris truncata, an Australian native C4 grass and a summer-fallow weed, which is common in no-till agriculture situations where herbicides are involved in crop management. To investigate the role of drought and increased atmospheric carbon dioxide (CO2) in determining weed growth, three trials were conducted using a ‘glyphosate-resistant’ and a ‘glyphosate-susceptible’ biotype. The first two trials tested the effect of herbicide (glyphosate) application on plant survival and growth under moisture stress and elevated CO2 respectively. A third trial investigated the effect on plant growth and reproduction under conditions of moisture stress and elevated CO2 in the absence of herbicide. In the first trial, water was withheld from half of the plants prior to application of glyphosate to all plants, and in the second trial plants were grown in either ambient (450 ppm) or elevated CO2 levels (750 ppm) prior to, and following, herbicide application. In both biotypes, herbicide effectiveness was reduced when plants were subjected to moisture stress or if grown in elevated CO2. Plant productivity, as measured by dry biomass per plant, was reduced with moisture stress, but increased with elevated CO2. In the third trial, growth rate, biomass and seed production were higher in the susceptible biotype compared to the resistant biotype. This suggests that a superior ability to resist herbicides may come at a cost to overall plant fitness. The results indicate that control of this weed may become difficult in the future as climatic conditions change. Supplementary information accompanies this paper at https://doi.org/10.1038/s41598-019-47237-x