Publications by authors named "Daniel J Browne"
Article Synopsis
- The proper collection, cryopreservation, thawing, and culture of peripheral blood mononuclear cells (PBMCs) are crucial for maintaining T cell viability and immunogenicity, yet current protocols are often inconsistently followed.
- A review of the literature reveals significant variations in technical processes that affect the quality of PBMCs, underscoring the need for more rigorous methodology and transparent reporting to promote reproducibility in research.
- The authors advocate for adopting the standardized operating procedures (SOPs) established by HANC to enhance consistency in PBMC processing, emphasizing detailed documentation of any variations to improve scientific reliability.
Article Synopsis
- The study evaluates the stability of twelve commonly used reference genes in RTqPCR analysis following Plasmodium yoelii sporozoite challenge and immunization in mice.
- It found significant expression changes in six of these genes due to the parasite challenge or immunization, indicating that not all reference genes are suitable for this type of analysis.
- SDHA and TBP were identified as stable reference genes, leading to the development of a robust RTqPCR protocol that can improve the consistency and reliability of malaria vaccine efficacy studies across different research settings.
Whole-blood-derived transcriptional profiling is widely used in biomarker discovery, immunological research, and therapeutic development. Traditional molecular and high-throughput transcriptomic platforms, including molecular assays with quantitative PCR (qPCR) and RNA-sequencing (RNA-seq), are dependent upon high-quality and intact RNA. However, collecting high-quality RNA from field studies in remote tropical locations can be challenging due to resource restrictions and logistics of post-collection processing.
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- Current immune assays are sensitive but require a lot of cells and expensive reagents, making them impractical for large-scale testing.
- A new RT-qPCR-based high-throughput screening assay has been developed, which significantly reduces the need for cell numbers and lowers costs by almost 90% while maintaining sensitivity.
- This optimized assay can accurately measure immune responses using as few as 50,000 PBMCs, making it ideal for situations where samples are limited and budgets are tight.
Novel antiinflammatory biologics shaped by parasite-host coevolution.
Stephanie M Ryan Roland Ruscher Wayne A Johnston Darren A Pickering Malcolm W Kennedy Daniel J Browne
Proc Natl Acad Sci U S A
September 2022
Article Synopsis
- Researchers explored using the immunomodulatory properties of helminth proteins, specifically by creating a library of recombinant proteins from hookworms.
- They identified 20 proteins that showed significant anti-inflammatory effects in a mouse model, which could lead to novel treatments for IBD and other inflammatory diseases.
Article Synopsis
- Nitrogen fertilizers, especially ammonium, are commonly used to boost banana production but can worsen Fusarium wilt, a major disease affecting banana plants.
- In a greenhouse study, increasing nitrogen rates correlated positively with the growth of non-inoculated banana plants, while plants inoculated with the disease showed increased severity and growth at intermediate nitrogen levels.
- The research revealed that nitrogen application influenced metabolic pathways in bananas and altered microbial diversity in the soil, with greater disease severity linked to reduced plant defense responses, especially when using ammonium fertilizers.
Article Synopsis
- The study demonstrates the effectiveness of a multiplex microsphere polymerase chain reaction (mmPCR) method for detecting both general and specific genetic targets in a single test.
- The mmPCR effectively identified various bacterial and fungal isolates using custom primers, achieving accurate results for both general classifications and specific traits such as antibiotic resistance.
- The findings suggest that mmPCR could enhance clinical diagnostics, though improvements in DNA extraction methods are needed for better sensitivity.
Article Synopsis
- Maternal choline supplementation in rats can help counteract some negative effects of alcohol exposure during pregnancy, particularly in terms of fetal growth and placental changes.
- In an experiment, pregnant rats were given different diets with varying levels of choline and alcohol to assess the impact on fetal and placental development.
- The results indicated that higher choline levels improved fetal weight and placental efficiency, especially when compared to alcohol exposure, while also causing alterations in DNA methylation patterns in placentas based on the offspring's sex.
Article Synopsis
- Reliable extraction of RNA from human PBMCs is essential for immunology research and clinical diagnostics, but it's challenging due to low sample availability and sensitivity issues.
- The study evaluates various RNA extraction and quantification kits, finding significant differences in RNA expression and establishing a reliable normalization strategy using absolute quantification based on cell numbers.
- An optimized RNA extraction and RT-qPCR protocol was developed that enables high sensitivity, sufficient for detecting specific immune responses with minimal PBMCs, making it a promising alternative to traditional protein assays.
Allogeneic bone marrow transplantation (BMT) provides curative therapy for leukemia via immunologic graft-versus-leukemia (GVL) effects. In practice, this must be balanced against life threatening pathology induced by graft-versus-host disease (GVHD). Recipient dendritic cells (DC) are thought to be important in the induction of GVL and GVHD.
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- Bloodstream infections pose a serious threat, especially to patients undergoing treatments like chemotherapy, where timely diagnosis is crucial due to the risks of febrile neutropenia.
- Traditional blood culture methods for diagnosing these infections can take hours to days, leading to the use of broad-spectrum antibiotics as an immediate response.
- The study introduces a rapid multiplexed microsphere-PCR assay that can differentiate between Gram-positive and Gram-negative bacteria in just 3.5 hours, potentially enhancing the speed and accuracy of infection treatment.