Objectives versus realities: Spatial,temporal, financial and social deficiencies in Australia’s public revegetation investment model

Past and continuing fragmentation and modification of ecosystems, as well as other threatening processes, cause ongoing biodiversity losses and species extinctions in Australia. At the same time as biodiversity declines, government funding for conservation and restoration is diminishing, leading to reduced action and greater reliance on private investment and community groups. In order to maintain and restore biodiverse ecosystems and the essential services they provide, both conservation of existing vegetation and habitat reconstruction are required. In this paper, we summarise the available data on planting area and cost from the Australian Government’s 20 Million Trees programme (2014–2020), the largest recent national‐scale revegetation incentives programme in Australia. We find that the current spatial scale of effort and investment in habitat reconstruction is insufficient to match the scale required to meet national conservation objectives. Furthermore, the funding rate ($/ha) and contracting arrangements are inadequate for the establishment of high‐quality self‐sustaining vegetation needed for the recovery of Australia’s threatened species and ecological communities. We estimate that the minimum amount of funding required for habitat reconstruction is at least five times higher than is provided for current national flagship programmes such as 20 Million Trees. We provide recommendations, designed to assist future habitat reconstruction programmes achieve their long‐term biodiversity objectives.     

Localization of the lipid probe 1,6-diphenyl-1,3,5 hexatriene (DPH) in intact cells by fluorescence microscopy

The lipophilic fluorescent probe DPH, generally used to determine the microviscosity of membrane lipids, has been visualized in intact cells by fluorescence microscopy. All lipid material of the cells, including cytoplasmic lipid droplets, was found to be labelled with DPH. The fluorescent signal from inside the cells contributes to a large extent to the total cell fluorescence. The results indicate that fluorescence polarization data obtained from intact cells, using DPH as probe, give information on the total lipid material of the cells rather than exclusive information on microviscosity and fluidity of plasma membranes of these cells, as has been repeatedly suggested.     

Crosstalk between small GTPases and polarity proteins in cell polarization

Cell polarization is crucial for the development of multicellular organisms, and aberrant cell polarization contributes to various diseases, including cancer. How cell polarity is established and how it is maintained remain fascinating questions. Conserved proteins of the partitioning defective (PAR), Scribble and Crumbs complexes guide the establishment of cell polarity in various organisms. Moreover, GTPases that regulate actin cytoskeletal dynamics have been implicated in cell polarization. Recent findings provide insights into polarization mechanisms and show intriguing crosstalk between small GTPases and members of polarity complexes in regulating cell polarization in different cellular contexts and cell types.     

T-helper 17 cell cytokines and interferon type I: partners in crime in systemic lupus erythematosus?

Introduction

A hallmark of systemic autoimmune diseases like systemic lupus erythematosus (SLE) is the increased expression of interferon (IFN) type I inducible genes, so-called IFN type I signature. Recently, T-helper 17 subset (Th17 cells), which produces IL-17A, IL-17F, IL-21, and IL-22, has been implicated in SLE. As CCR6 enriches for Th17 cells, we used this approach to investigate whether CCR6⁺ memory T-helper cells producing IL-17A, IL-17F, IL-21, and/or IL-22 are increased in SLE patients and whether this increase is related to the presence of IFN type I signature.

Methods

In total, 25 SLE patients and 15 healthy controls (HCs) were included. SLE patients were divided into IFN type I signature-positive (IFN⁺) (n = 16) and negative (IFN^-) (n = 9) patients, as assessed by mRNA expression of IFN-inducible genes (IFIGs) in monocytes. Expression of IL-17A, IL-17F, IL-21, and IL-22 by CD4⁺CD45RO⁺CCR6⁺ T cells (CCR6⁺ cells) was measured with flow cytometry and compared between IFN⁺, IFN^- patients and HCs.

Results

Increased percentages of IL-17A and IL-17A/IL-17F double-producing CCR6⁺ cells were observed in IFN⁺ patients compared with IFN^- patients and HCs. IL-17A and IL-17F expression within CCR6⁺ cells correlated significantly with IFIG expression. In addition, we found significant correlation between B-cell activating factor of the tumor necrosis family (BAFF)–a factor strongly correlating with IFN type I - and IL-21 producing CCR6⁺ cells.

Conclusions

We show for the first time higher percentages of IL-17A and IL-17A/IL-17F double-producing CCR6⁺ memory T-helper cells in IFN⁺ SLE patients, supporting the hypothesis that IFN type I co-acts with Th17 cytokines in SLE pathogenesis.     

Bromate induces loss of heterozygosity in the thymidine kinase gene of L5178Y/Tk(+/-)-3.7.2C mouse lymphoma cells

Potassium bromate (KBrO(3)) induces DNA damage and tumors in mice and rats, but is a relatively weak mutagen in microbial assays and the in vitro mammalian Hprt assay. Concern that there may be a human health risk associated with bromate, a disinfectant by-product of ozonation, has accompanied the increasing use of ozonation as an alternative to chlorination for treatment of drinking water. In this study, we have evaluated the mutagenicity of KBrO(3) and sodium bromate (NaBrO(3)) in the Tk gene of mouse lymphoma cells. In contrast to the weak mutagenic activity seen in the previous studies, bromate induced a mutant frequency of over 100 x 10(-6) at 0.6mM with minimal cytotoxicity (70-80% survival) and over 1300 x 10(-6) at 3mM ( approximately 10% survival). The increase in the Tk mutant frequency was primarily due to the induction of small colony of Tk mutants. Loss of heterozygosity (LOH) analysis of 384 mutants from control and 2.7 mM KBrO(3)-treated cells showed that almost all (99%) bromate-induced mutants resulted from LOH, whereas in the control cultures 77% of the Tk mutants were LOH. Our results suggest that bromate is a potent mutagen in the Tk gene of mouse lymphoma cells, and the mechanism of action primarily involves LOH. The ability of the mouse lymphoma assay to detect a wider array of mutational events than the microbial or V79 Hprt assays may account for the potent mutagenic response.     

A bacterial population study of commercialized wastewater inoculants

AIMS: To assess the bacterial diversity and safety of wastewater inoculants, which are commercially available products used to improve the aerobic digestion processes of the domestic waste compost in the septic tank. METHODS AND RESULTS: Eighteen wastewater inoculants were analysed on nonselective and selective media and the cultivable bacteria were identified. In all wastewater inoculants, the number of CFUs were between 10(4) and 10(7) g(-1) powder on nonselective media and Bacillus was the predominant cultivable genus. Culture-independent molecular methods such as sequencing of 16S rRNA clone libraries and denaturating gradient gel electrophoresis demonstrated the high prevalence of interfering chloroplast 16S rRNA from plant material and the presence of Bacillus spp. Only after selective enrichments and cultivation, the presence of one pathogenic strain (Klebsiella pneumoniae subsp. pneumoniae) and one opportunistic strain of (Enterobacter cloacae) bacteria were detected in six different products. CONCLUSION: The predominant cultivable species of the wastewater inoculants were Bacillus spp. and after enrichment six products were found to contain opportunistic or pathogenic strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of opportunistic pathogenic strains in the inoculants might represent a risk for immunocompromised, the elderly or children. A clear labelling should therefore be displayed on the product.     

A marker-assisted backcross approach for developing submergence-tolerant rice cultivars

Submergence stress regularly affects 15 million hectares or more of rainfed lowland rice areas in South and Southeast Asia. A major QTL on chromosome 9, Sub1, has provided the opportunity to apply marker assisted backcrossing (MAB) to develop submergence tolerant versions of rice cultivars that are widely grown in the region. In the present study, molecular markers that were tightly linked with Sub1, flanking Sub1, and unlinked to Sub1 were used to apply foreground, recombinant, and background selection, respectively, in backcrosses between a submergence-tolerant donor and the widely grown recurrent parent Swarna. By the BC₂F₂ generation a submergence tolerant plant was identified that possessed Swarna type simple sequence repeat (SSR) alleles on all fragments analyzed except the tip segment of rice chromosome 9 that possessed the Sub1 locus. A BC₃F₂ double recombinant plant was identified that was homozygous for all Swarna type alleles except for an approximately 2.3–3.4 Mb region surrounding the Sub1 locus. The results showed that the mega variety Swarna could be efficiently converted to a submergence tolerant variety in three backcross generations, involving a time of two to three years. Polymorphic markers for foreground and recombinant selection were identified for four other mega varieties to develop a wider range of submergence tolerant varieties to meet the needs of farmers in the flood-prone regions. This approach demonstrates the effective use of marker assisted selection for a major QTL in a molecular breeding program. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Development of Real-Time PCR Methods for the Detection of Bacterial Meningitis Pathogens without DNA Extraction

Neisseria meningitidis (Nm), Haemophilus influenzae (Hi), and Streptococcus pneumoniae (Sp) are the lead causes of bacterial meningitis. Detection of these pathogens from clinical specimens using traditional real-time PCR (rt-PCR) requires DNA extraction to remove the PCR inhibitors prior to testing, which is time consuming and labor intensive. In this study, five species-specific (Nm-sodC and -ctrA, Hi-hpd#1 and -hpd#3 and Sp-lytA) and six serogroup-specific rt-PCR tests (A, B, C, W, X, Y) targeting Nm capsular genes were evaluated in the two direct rt-PCR methods using PerfeCTa and 5x Omni that do not require DNA extraction. The sensitivity and specify of the two direct rt-PCR methods were compared to TaqMan traditional rt-PCR, the current standard rt-PCR method for the detection of meningitis pathogens. The LLD for all 11 rt-PCR tests ranged from 6,227 to 272,229 CFU/ml for TaqMan, 1,824–135,982 for 5x Omni, and 168–6,836 CFU/ml for PerfeCTa. The diagnostic sensitivity using TaqMan ranged from 89.2%-99.6%, except for NmB-csb, which was 69.7%. For 5x Omni, the sensitivity varied from 67.1% to 99.8%, with three tests below 90%. The sensitivity of these tests using PerfeCTa varied from 89.4% to 99.8%. The specificity ranges of the 11 tests were 98.0–99.9%, 97.5–99.9%, and 92.9–99.9% for TaqMan, 5x Omni, and PerfeCTa, respectively. PerfeCTa direct rt-PCR demonstrated similar or better sensitivity compared to 5x Omni direct rt-PCR or TaqMan traditional rt-PCR. Since the direct rt-PCR method does not require DNA extraction, it reduces the time and cost for processing CSF specimens, increases testing throughput, decreases the risk of cross-contamination, and conserves precious CSF. The direct rt-PCR method will be beneficial to laboratories with high testing volume.