Single-molecule fluorescence imaging of DNA maintenance protein binding dynamics and activities on extended DNA.

Defining the molecular mechanisms by which genome maintenance proteins dynamically associate with and process DNA is essential to understand the potential avenues by which these stabilizing mechanisms are disrupted. Single-molecule fluorescence imaging (SMFI) of protein dynamics on extended DNA has greatly expanded our ability to accomplish this, as it captures singular biomolecular interactions - in all their complexity and diversity - without relying on ensemble-averaging of bulk protein activity as most traditional biochemical techniques must do. In this review, we discuss how SMFI studies with extended DNA have substantially contributed to genome stability research over the past two years.

High-speed AFM imaging reveals DNA capture and loop extrusion dynamics by cohesin-NIPBL.

3D chromatin organization plays a critical role in regulating gene expression, DNA replication, recombination, and repair. While initially discovered for its role in sister chromatid cohesion, emerging evidence suggests that the cohesin complex (SMC1, SMC3, RAD21, and SA1/SA2), facilitated by NIPBL, mediates topologically associating domains and chromatin loops through DNA loop extrusion. However, information on how conformational changes of cohesin-NIPBL drive its loading onto DNA, initiation, and growth of DNA loops is still lacking.

Single-molecule imaging of genome maintenance proteins encountering specific DNA sequences and structures.

DNA repair pathways are tightly regulated processes that recognize specific hallmarks of DNA damage and coordinate lesion repair through discrete mechanisms, all within the context of a three-dimensional chromatin landscape. Dysregulation or malfunction of any one of the protein constituents in these pathways can contribute to aging and a variety of diseases. While the collective action of these many proteins is what drives DNA repair on the organismal scale, it is the interactions between individual proteins and DNA that facilitate each step of these pathways.

Densely methylated DNA traps Methyl-CpG-binding domain protein 2 but permits free diffusion by Methyl-CpG-binding domain protein 3.

The methyl-CpG-binding domain 2 and 3 proteins (MBD2 and MBD3) provide structural and DNA-binding function for the Nucleosome Remodeling and Deacetylase (NuRD) complex. The two proteins form distinct NuRD complexes and show different binding affinity and selectivity for methylated DNA. Previous studies have shown that MBD2 binds with high affinity and selectivity for a single methylated CpG dinucleotide while MBD3 does not.

An estimate of phthalate exposure among pregnant women living in Trujillo, Peru.

Phthalates are a group of phthalic acid esters which are used as plasticizers and additives. In laboratory animals, several phthalates are known endocrine disruptors. Several studies have described phthalate exposure in the United States and developed countries but little is known about phthalate exposure in the developing world, particularly during pregnancy.

Immunological and pathological changes in the placenta during infection with Listeria monocytogenes in pregnant guinea pigs.

Exposure to Listeria monocytogenes during pregnancy can result in spontaneous abortion and stillbirths; however, the mechanisms are unknown. Our objective was to determine the effects of infection on specific inflammatory and anti-inflammatory cytokine mRNA expression and apoptosis in the placenta after infection with L. monocytogenes.

Listeria monocytogenes infection in pregnant guinea pigs is associated with maternal liver necrosis, a decrease in maternal serum TNF-alpha concentrations, and an increase in placental apoptosis.

Stillbirths and spontaneous abortions can result when pregnant women are exposed to the food borne pathogen, Listeria monocytogenes. Fetuses and neonates account for one-third of the 2500 cases annually. The objectives were to determine the dose dependent trends of immunological and pathological effects in pregnant guinea pigs after infection with L.

A processing plant persistent strain of Listeria monocytogenes crosses the fetoplacental barrier in a pregnant guinea pig model.

The foodborne pathogen Listeria monocytogenes can cause infection in immunocompromised humans and in the fetuses of pregnant women. We have demonstrated that one group of genetically similar L. monocytogenes strains (random amplified polymorphic DNA [RAPD] type 9) dominate and persist in several independent fish processing plants.

Dose-response of Listeria monocytogenes after oral exposure in pregnant guinea pigs.

Listeriosis, a severe disease that results from exposure to the foodborne pathogen Listeria monocytogenes, is responsible for approximately 2500 illnesses and 500 deaths in the United States each year. Pregnant women are 20 times more likely to develop listeriosis than the general population, with adverse pregnancy outcomes that include spontaneous abortions, stillbirths, and neonatal meningitis. The objective of this study was to determine an infective dose that resulted in stillbirths and infectivity of selected tissues in pregnant guinea pigs.

Alliance formation with parents: limit-setting and the effect of mandated reporting.

The ability of parents to cooperate in their children's treatment is not fixed, but is a potential that evolves in a clinical relationship. Such clinical work includes a hierarchy of limit-setting, ranging from education to legal intervention. The experience of an inpatient child psychosomatic service indicates that such limit-setting was relevant in more than 50% of cases, and served to enhance the alliance with parents and children.