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Organic fertilizers and microbial biofertilizers are now widely recognized to effectively complement traditional mineral fertilizers for plant growth. The present study shows that bio-organic fertilizers can be enhanced by the addition of functional plant-growth-promoting rhizobacteria (PGPR) that provide additional benefits to plants. We hypothesized that not all beneficial soil bacteria are functional in different farm soils and plant varieties; hence, the most effective PGPR that are suitable to each farm’s individual cropping conditions were selected. Five different field soils and their respective crops were tested for compatibility with six microbial biofertilizers (including three new bacterial strains) to supplement a commercially available bio-organic fertilizer. In pot trials with lucerne plants, four out of the six microbial treatments led to significant (p < 0.05) growth promotion benefits (up to 79.8% more leaves and dry weight) compared to mock-treated or bio-organic fertilizer-only-treated control plants. A trial with industrial hemp demonstrated that compatibility with PGPR occurs in a cultivar-specific manner, leading to growth promotion ranging from −3.4% to 68.9%, with each cultivar displaying a preference for a different PGPR. Finally, pot trials with Rhodes grass and two different soils demonstrated high yield increases compared to control plants, with Bacillus amyloliquefaciens 33YE being most effective for one soil and Bacillus velezensis UQ9000N/Pseudomonas lini SMX2 for the other soil. Yield advantages reduced after several cuts of grass, but a repeat biofertilizer treatment at 69 days after the initial treatment restored high yield advantages, with the same PGPR again being most effective. These results demonstrate the importance of customization of microbial inoculants to identify the most compatible PGPR–cultivar–soil interaction. The customization of microbial biofertilizers to soils and plant cultivars, combined with complementary fertilizer applications, can potentially lead to more reliable and more sustainable agricultural practices.

The increase in global population and industrial development has led to a significant release of organic and inorganic pollutants into water streams, threatening human health and ecosystems. Microalgae, encompassing eukaryotic protists and prokaryotic cyanobacteria, have emerged as a sustainable and cost-effective solution for removing these pollutants and mitigating carbon emissions. Various microalgae species, such as C. vulgaris, P. tricornutum, N. oceanica, A. platensis, and C. reinhardtii, have demonstrated their ability to eliminate heavy metals, salinity, plastics, and pesticides. Synthetic biology holds the potential to enhance microalgae-based technologies by broadening the scope of treatment targets and improving pollutant removal rates. This review provides an overview of the recent advances in the synthetic biology of microalgae, focusing on genetic engineering tools to facilitate the removal of inorganic (heavy metals and salinity) and organic (pesticides and plastics) compounds. The development of these tools is crucial for enhancing pollutant removal mechanisms through gene expression manipulation, DNA introduction into cells, and the generation of mutants with altered phenotypes. Additionally, the review discusses the principles of synthetic biology tools, emphasizing the significance of genetic engineering in targeting specific metabolic pathways and creating phenotypic changes. It also explores the use of precise engineering tools, such as CRISPR/Cas9 and TALENs, to adapt genetic engineering to various microalgae species. The review concludes that there is much potential for synthetic biology based approaches for pollutant removal using microalgae, but there is a need for expansion of the tools involved, including the development of universal cloning toolkits for the efficient and rapid assembly of mutants and transgenic expression strains, and the need for adaptation of genetic engineering tools to a wider range of microalgae species.

Tomer Snir,1 Ilya Kirgner,2 Nachum Nesher,3 Yaacov Abramov,3 Marina Kolodii,3 Sharbel Azzam,3 Michael Peer3 1Medical Students Elective Program, University of Milan, Milan, Italy; 2Blood Bank, Ichilov Medical Center, Tel Aviv, Israel; 3Department of Thoracic Surgery, Ichilov Medical Center, Tel Aviv, IsraelCorrespondence: Michael Peer, Tel +972-3-6972021, Fax +972-3 - 6973235, Email michaelpe@tlvmc.gov.ilBackground: Blood transfusion rates during surgery and hospitalization for thoracic surgery vary from 16% to 55%. The religious beliefs of Jehovah’s Witnesses (JW) permit medical and surgical procedures but exclude the use of blood and blood products. Performing major pulmonary resection without the possibility of compensating for blood loss is a daunting challenge that few surgeons are willing to meet.Methods: The clinical and surgical data on two JW patients who underwent major pulmonary resections for non-small cell lung carcinoma (NSCLC) in Tel Aviv Medical Center between the years 2019 and 2020 were retrieved from the departmental databases and analyzed for the requirement and consumption of blood products. The patients were a 70-year-old female and a 49-year-old man that have underwent a completion right upper lobe lobectomy and a left lower lobe lobectomy, respectively.Results: None required blood transfusions and the surgeries were successful, demonstrating that it is possible to successfully perform “bloodless” major pulmonary resection while respecting the limitations set by the patient’s religious beliefs.Conclusion: This concept paves the way for the consideration of major pulmonary resection for patients who are currently denied such procedures due to religious restrictions or to their being high-risk surgical candidates, when resources are limited or lacking.Keywords: bloodless thoracic surgery, lobectomy, Jehovah’s Witnesses

Rhizosphere bacteria can provide multiple benefits to plants, including increased nutrient supply, pathogen/disease control, and abiotic stress tolerance, but results from pot trials do not always translate to field conditions. This study tested whether rhizosphere biocontrol bacteria can also provide plant growth promotion and how benefits can be provided at a commercial farm. Commercial lettuce seeds and plants were treated with rhizosphere biocontrol bacteria Bacillus velezensis UQ9000N, B. amyloliquefaciens 33YE, Brevibacillus laterosporus 4YE, and Pseudomonas azotoformans UQ4510An. 33YE increased the head diameter, plant height, and fresh weight of the Green Moon cultivar, while 33YE, UQ4510An, and UQ9000N increased the fresh and dry weight of Liston, a more heat-tolerant cultivar, via a single seed treatment or repeat root treatments under nursery and field conditions across different inoculation schedules and growth stages. Significant growth promotion was also demonstrated when inoculating field plants after transplanting (in particular for 33YE). Applications of these microbial biostimulants to lettuce seeds or plantlets potentially enable earlier transplanting and earlier harvests. Repeat inoculations using irrigation water and long-lasting formulations may further advance the benefits of these biostimulants as microbial biofertilisers for plant growth promotions in the field.

Bacterial biocontrol agent/s (BCAs) against plant diseases are eco-friendly and sustainable options for profitable agricultural crop production. Specific beneficial strains of Bacillus subtilis are effective in controlling many fungal diseases including Alternaria blight caused by a notorious pathogen “Alternaria solani”. In the present study, the biocontrol attributes of a newfangled strain of B. subtilis (BS-01) have been investigated and its bioactive compounds were also identified against A. solani. The volatile organic compounds (VOCs) produced by BS-01 in organic solvents viz., n-hexane, dichloromethane, and ethyl acetate were extracted and their antifungal efficacy has evaluated against A. solani. Also, the preventive and curative biocontrol method to reduce the fungal load of A. solani was estimated by both foliar and seed applications on infected tomato (Solanum lycopersicum) plants as determined by quantitative PCR assays. Growth chamber bioassay revealed that both foliar and seed application of BS-01 on tomato plants previously or subsequently infected by A. solani significantly reduced the pathogen load on inoculated tomato foliage. Results showed that antifungal bioassays with various concentrations (10-100 mg mL-1) of extracted metabolites produced by BS-01 in ethyl acetate fraction showed the highest inhibition in fungal biomass (extracellular metabolites: 69-98% and intracellular metabolites: 48-85%) followed by n-hexane (extracellular metabolites: 63-88% and intracellular metabolites: 35-62%) and dichloromethane (extracellular metabolites: 41-74% and intracellular metabolites: 42-70%), respectively. The extracted volatile compounds of BS-01 were identified via GC-MS analysis and were found in great proportions in the organic fractions as major potent antifungal constituents including triphenylphosphine oxide; pyrrolo[1,2-a] pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl); pyrrolo[1,2-a] pyrazine-1,4-dione, hexahydro-3-(phenylmethyl); n-hexadecanoic acid; n-tridecan-1-ol; octadecane; octadecanoic acid; eicosane and dodecyl acrylate. Separate or mixture of these bioactive VOCs had the potential to mitigate the tomato early blight disease severity in the field that would act as a sustainable plant protection strategy to generate profitable tomato production.

Abstract Background Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor that facilitates the adaptation of cancer cells to hypoxic conditions and may be prognostic of breast cancer recurrence. We evaluated the association of HIF-1α expression with breast cancer recurrence, and its association with timing of breast cancer recurrence. Methods In this population-based case-control study, we included women diagnosed with stage I–III breast cancer between 1985 and 2001, aged 35–69 years, registered in the Danish Breast Cancer Group. We identified 541 cases of breast cancer recurrence among women with estrogen receptor (ER)-positive disease who were treated with tamoxifen for at least 1 year (ER+ TAM+). We also enrolled 300 breast cancer recurrence cases among women with ER-negative disease, not treated with tamoxifen, who survived at least 1 year (ER−/TAM−). Controls were recurrence-free breast cancer patients at the time of case diagnosis, matched to recurrence cases on ER/TAM status, date of surgery, menopausal status, cancer stage, and county of residence. Expression of HIF-1α was measured by immunohistochemistry on tissue microarrays. We fitted logistic regression models to compute odds ratios (ORs) and 95% confidence intervals (CIs) associating HIF-1α expression with recurrence, and with timing of recurrence. Results HIF-1α expression was observed in 23% of cases and 20% of controls in the ER+/TAM+ stratum, and in 47% of cases and 48% of controls in the ER−/TAM− stratum. We observed a near-null association between HIF-1α expression in both ER/TAM groups (ER+/TAM+ OR = 1.21, 95%CI 0.88, 1.67 and ER−/TAM− OR = 0.97, 95%CI 0.68, 1.39). HIF-1α expression was not associated with time to recurrence among women in the ER+/TAM+ stratum, but was associated with early recurrence among women in the ER−/TAM− stratum. Conclusion In this study, HIF-1α expression was not associated with breast cancer recurrence overall but may be associated with early recurrence among women diagnosed with ER− breast cancer.

We hypothesized that environmental microbiomes contain a wide range of bacteria that produce yet uncharacterized antimicrobial compounds (AMCs) that can potentially be used to control pathogens. Over 600 bacterial strains were isolated from soil and food compost samples, and 68 biocontrol bacteria with antimicrobial activity were chosen for further studies based on inhibition assays against a wide range of food and plant pathogens. For further characterization of the bioactive compounds, a new method was established that used living pathogens in a liquid culture to stimulate bacteria to produce high amounts of AMCs in bacterial supernatants. A peptide gel electrophoresis microbial inhibition assay was used to concurrently achieve size separation of the antimicrobial peptides. Fifteen potential bioactive peptides were then further characterized by tandem MS, revealing cold-shock proteins and 50S ribosomal proteins. To identify non-peptidic AMCs, bacterial supernatants were analyzed by HPLC followed by GC/MS. Among the 14 identified bioactive compounds, 3-isobutylhexahydropyrrolo[1,2-a]pyrazine-1,4-dione and 2-acetyl-3-methyl-octahydropyrrolo[1,2-a]piperazine-1,4-dione were identified as new AMCs. Our work suggests that antimicrobial compound production in microbes is enhanced when faced with a threat from other microorganisms, and that this approach can rapidly lead to the development of new antimicrobials with the potential for upscaling.

Organic fertilizers and microbial biofertilizers are now widely recognized to effectively complement traditional mineral fertilizers for plant growth. The present study shows that bio-organic fertilizers can be enhanced by the addition of functional plant-growth-promoting rhizobacteria (PGPR) that provide additional benefits to plants. We hypothesized that not all beneficial soil bacteria are functional in different farm soils and plant varieties; hence, the most effective PGPR that are suitable to each farm's individual cropping conditions were selected. Five different field soils and their respective crops were tested for compatibility with six microbial biofertilizers (including three new bacterial strains) to supplement a commercially available bio-organic fertilizer. In pot trials with lucerne plants, four out of the six microbial treatments led to significant (p < 0.05) growth promotion benefits (up to 79.8% more leaves and dry weight) compared to mock-treated or bio-organic fertilizer-only-treated control plants. A trial with industrial hemp demonstrated that compatibility with PGPR occurs in a cultivar-specific manner, leading to growth promotion ranging from −3.4% to 68.9%, with each cultivar displaying a preference for a different PGPR. Finally, pot trials with Rhodes grass and two different soils demonstrated high yield increases compared to control plants, with Bacillus amyloliquefaciens 33YE being most effective for one soil and Bacillus velezensis UQ9000N/Pseudomonas lini SMX2 for the other soil. Yield advantages reduced after several cuts of grass, but a repeat biofertilizer treatment at 69 days after the initial treatment restored high yield advantages, with the same PGPR again being most effective. These results demonstrate the importance of customization of microbial inoculants to identify the most compatible PGPR–cultivar–soil interaction. The customization of microbial biofertilizers to soils and plant cultivars, combined with complementary fertilizer applications, can potentially lead to more reliable and more sustainable agricultural practices. [ABSTRACT FROM AUTHOR]

Purpose: Breast cancer has an average 10-year relative survival reaching 84%. This favorable survival is due, in part, to the introduction of biomarker-guided therapies. We estimated the population-level effect of the introduction of two adjuvant therapies--tamoxifen and trastuzumab--on recurrence using the trend-in-trend pharmacoepidemiologic study design. Methods: We ascertained data on women diagnosed with nonmetastatic breast cancer who were registered in the Danish Breast Cancer Group clinical database. We used the trend-in-trend design to estimate the population-level effect of the introduction of (1) tamoxifen for postmenopausal women with estrogen receptor (ER)-positive breast cancer in 1982, (2) tamoxifen for premenopausal women diagnosed with ER-positive breast cancer in 1999, and (3) trastuzumab for women <60 years diagnosed with human epidermal growth factor receptor 2-positive breast cancer in 2007. Results: For the population-level effect of the introduction of tamoxifen among premenopausal women diagnosed with ER-positive breast cancer in 1999, the risk of recurrence decreased by nearly one-half (OR = 0.52), consistent with evidence from clinical trials; however, the estimate was imprecise (95% confidence interval [CI] = 0.25, 1.85). We observed an imprecise association between tamoxifen use and recurrence from the time it was introduced in 1982 (OR = 1.24 95% CI = 0.46, 5.11), inconsistent with prior knowledge from clinical trials. For the introduction of trastuzumab in 2007, the estimate was also consistent with trial evidence, though imprecise (OR = 0.51; 95% CI = 0.21, 22.4). Conclusions: We demonstrated how novel pharmacoepidemiologic analytic designs can be used to evaluate the routine clinical care and effectiveness of therapeutic advancements in a population-based setting while considering some limitations of the approach. [ABSTRACT FROM AUTHOR]

Background: Na+,HCO3–-cotransporter NBCn1/Slc4a7 accelerates murine breast carcinogenesis. Lack of specific pharmacological tools previously restricted therapeutic targeting of NBCn1 and identification of NBCn1-dependent functions in human breast cancer. Methods: We develop extracellularly-targeted anti-NBCn1 antibodies, screen for functional activity on cells, and evaluate (a) mechanisms of intracellular pH regulation in human primary breast carcinomas, (b) proliferation, cell death, and tumor growth consequences of NBCn1 in triple-negative breast cancer, and (c) association of NBCn1-mediated Na+,HCO3–-cotransport with human breast cancer metastasis. Results: We identify high-affinity (KD ≈ 0.14 nM) anti-NBCn1 antibodies that block human NBCn1-mediated Na+,HCO3–-cotransport in cells, without cross-reactivity towards human NBCe1 or murine NBCn1. These anti-NBCn1 antibodies abolish Na+,HCO3–-cotransport activity in freshly isolated primary organoids from human breast carcinomas and lower net acid extrusion effectively in primary breast cancer tissue from patients with macrometastases in axillary lymph nodes. Inhibitory anti-NBCn1 antibodies decelerate tumor growth in vivo by ~50% in a patient-derived xenograft model of triple-negative breast cancer and pH-dependently reduce colony formation, cause G2/M-phase cell cycle accumulation, and increase apoptosis of metastatic triple-negative breast cancer cells in vitro. Conclusions: Inhibitory anti-NBCn1 antibodies block net acid extrusion in human breast cancer tissue, particularly from patients with disseminated disease, and pH-dependently limit triple-negative breast cancer growth. [ABSTRACT FROM AUTHOR]

This paper aims to model and predict the mechanical properties of acrylonitrile-butadiene-styrene parts manufactured by fused deposition modelling. An L27 Taguchi array was used to associate the peculiar parameters (nozzle diameter, layer height, fill density, printing velocity, raster orientation and infill pattern) to perform experiments. A three-point bending test was executed on all samples in accordance with the ASTM D7264 standard, aimed to obtain Flexural strength and the 0.2% offset yield strength. An artificial neural network multi-parameter regression model was formed and then the model of the input-output relations developed by this network was optimised by genetic algorithm. This modelling and optimisation suggest a direct relation between choosing process parameters correctly and enhancing performance fused deposition modelling. This method provides optimal solutions for getting allotted output values. Infill pattern plays a critical role in providing strength. Infill patterns can be recommended for desired output within constraints of input factors. [ABSTRACT FROM AUTHOR]