Maternal immune stimulation during pregnancy shapes the immunological phenotype of offspring.

Epidemiological studies have associated infection during pregnancy with increased risk of neurodevelopmental disorders in children, which is modeled in rodents by stimulating the immune system of pregnant dams with microorganisms or their mimics, such as poly(I:C) or LPS. In two prenatal mouse models, we show that in utero exposure of the fetus to cytokines/inflammatory mediators elicited by maternal immune stimulation with poly(I:C) yields offspring that exhibit a proinflammatory phenotype due to alterations in developmental programming of their immune system. Changes in the innate and adaptive immune elements of these pro-inflammatory offspring result in more robust responses following exposure to immune stimuli than those observed in control offspring from PBS-injected pregnant dams.

Effects of temperature, surfactants and skin location on the dermal penetration of haloacetonitriles and chloral hydrate.

Dermal exposure has been recognized as an important contributor to the total internal dose to disinfection-by-products (DBPs) in water. However, the effect of the use of surfactants, water temperature and area of the body exposed to DBPs on their dermal flux has not been characterized and was the focus of the present study using an in-vitro system. The dermal flux of mg/l concentrations of haloacetonitriles and chloral hydrate (CH), important cytotoxic DBPs, increased by approximately 50% to 170% with increasing temperature from 25 °C to 40 °C.

Maternal immune stimulation during pregnancy affects adaptive immunity in offspring to promote development of TH17 cells.

Behavioral abnormalities in offspring of murine dams that receive immune stimulation with (poly)I:C during pregnancy are well-documented. In this prenatal model, (poly)I:C-induced maternal cytokines, particularly IL-6, appear involved in the etiology of the behavioral abnormalities. While much has been published on the abnormal behaviors of offspring in this model, much less is known about how maternal immune stimulation affects the adaptive immune system of the offspring, and its possible role in the observed pathophysiology.

Preparation of embryos for electron microscopy of the Drosophila embryonic heart tube.

The morphogenesis of the Drosophila embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell shape changes during embryonic development. One of the challenges faced in studying this structure is that the lumen of the heart tube, as well as the membrane features that are crucial to heart tube formation, are difficult to visualize in whole mount embryos, due to the small size of the heart tube and intra-lumenal space relative to the embryo. The use of transmission electron microscopy allows for higher magnification of these structures and gives the advantage of examining the embryos in cross section, which easily reveals the size and shape of the lumen.

Regulation of miRNA expression by Src and contact normalization: effects on nonanchored cell growth and migration.

Transformation by the Src tyrosine kinase (Src) promotes nonanchored cell growth and migration. However, nontransformed cells can force Src-transformed cells to assume a normal morphology and phenotype by a process called 'contact normalization'. It has become clear that microRNA (miRNA) can affect tumorigenesis by targeting gene products that direct cell growth and migration.

A simple method for isolating disomic strains of Saccharomyces cerevisiae.

A simple method to select disomic (N + 1) strains that should be applicable for almost any chromosome in Saccharomyces cerevisiae is presented. A diploid heterozygous for a KanMX knock-out mutation in an essential gene is sporulated and viable geneticin (G418)-resistant colonies selected. Disomic products of a missegregation or non-disjunction event containing a copy of both the wild-type essential gene and its complementary KanMX knock-out allele make up most of the viable colonies.

The plant homeodomain fingers of fission yeast Msc1 exhibit E3 ubiquitin ligase activity.

The DNA damage checkpoint pathway governs how cells regulate cell cycle progression in response to DNA damage. A screen for suppressors of a fission yeast chk1 mutant defective in the checkpoint pathway identified a novel Schizosaccharomyces pombe protein, Msc1. Msc1 contains 3 plant homeodomain (PHD) finger motifs, characteristically defined by a C4HC3 consensus similar to RING finger domains.

New modes of exchanger regulation: physiological implications.

Exchange activity is regulated principally by cytosolic Na+, Ca2+, and PIP2. However, the properties of these modes of regulation that have emerged from excised patch studies appear to be poorly suited to regulating exchange activity on a beat-to-beat basis. Here we summarize recent findings from our lab indicating that (a) allosteric activation by Ca2+ exhibits hysteresis, (b) elevated concentrations of cytosolic Na+ induce a mode of activity that no longer requires regulatory Ca2+ activation, and (c) the requirement for PIP2 is reduced or eliminated after allosteric Ca2+ activation.

In vivo analysis of synaptonemal complex formation during yeast meiosis.

During meiotic prophase a synaptonemal complex (SC) forms between each pair of homologous chromosomes and is believed to be involved in regulating recombination. Studies on SCs usually destroy nuclear architecture, making it impossible to examine the relationship of these structures to the rest of the nucleus. In Saccharomyces cerevisiae the meiosis-specific Zip1 protein is found throughout the entire length of each SC.

Regulation of the multifunctional Ca2+/calmodulin-dependent protein kinase II by the PP2C phosphatase PPM1F in fibroblasts.

The regulation of the multifunctional calcium/calmodulin dependent protein kinase II (CaMKII) by serine/threonine protein phosphatases has been extensively studied in neuronal cells; however, this regulation has not been investigated previously in fibroblasts. We cloned a cDNA from SV40-transformed human fibroblasts that shares 80% homology to a rat calcium/calmodulin-dependent protein kinase phosphatase that encodes a PPM1F protein. By using extracts from transfected cells, PPM1F, but not a mutant (R326A) in the conserved catalytic domain, was found to dephosphorylate in vitro a peptide corresponding to the auto-inhibitory region of CaMKII.

Inhalation exposure to haloacetic acids and haloketones during showering.

Inhalation exposure to haloacetic acids (HAAs) and haloketones (HKs) in contaminated drinking water occurs during showering. The size distribution of the aerosols generated by a shower was determined using an eight size-range particle counter, which measured particles from 0.1 to >2 microm.

Effects of hemispheric lateralization and site specificity on immune alterations induced by kindled temporal lobe seizures.

The effects of kindled seizures elicited from sites in the left and right temporal lobes on mitogen-induced proliferation (LPS, Con A, PHA) and induction of representative TH1 (IFN-gamma) and TH2 (IL-10, IL-4) cytokines were determined in activated rat splenocytes. With reference to cell proliferation, the changes depended on the hemispheric side and location of kindling. Kindling of the left side mediated significant increase in cell proliferation by LPS.

In vivo transcriptional analysis of the spliced leader RNA gene in the trypanosomatid Leptomonas seymouri.

Gene expression in all organisms requires the direct and indirect interaction of multiple proteins with specific DNA sequence elements. Using the monogenetic trypanosomatid, Leptomonas seymouri, we investigated the cis- and trans-acting components that determine expression of a central trypanosomatid RNA, the spliced leader (SL) RNA. Using base substitution mutagenesis and DNA transfection assays, we determined that the SL RNA gene promoter lies exclusively up-stream from the transcription initiation site.

Distribution of anionic sites on microvascular endothelium of the chick chorioallantoic membrane.

It is generally accepted that luminal surfaces of adult microvascular endothelia present an anionic barrier that limits passage of anionic macromolecules. To assess the ontogeny of the barrier, temporal and spatial expression of endothelial anionic sites was evaluated in the chorioallantoic membrane of chicken embryos from days 4.5 to 18 of incubation.

DNA replication fork pause sites dependent on transcription.

Replication fork pause (RFP) sites transiently arresting replication fork movement were mapped to transfer RNA (tRNA) genes of Saccharomyces cerevisiae in vivo. RFP sites are polar, stalling replication forks only when they oppose the direction of tRNA transcription. Mutant tRNA genes defective in assembly of transcription initiation complexes and a temperature-sensitive RNA polymerase III mutant (rpc160-41) defective in initiation of transcription do not stall replication forks, suggesting that transcription is required for RFP activity.