Enteric Glia Exhibit P 2U Receptors that Increase Cytosolic Calcium by a Phospholipase C-Dependent Mechanism

Calcium signaling in fura-2 acetoxymethyl ester-loaded enteric glia was investigated in response to neuroligands; responses to ATP were studied in detail. Carbachol (1 m M ), glutamate (100 µ M ), norepinephrine (10 µ M ), and substance P (1 µ M ) did not increase the intracellular calcium concentration ([Ca 2+ ] i ) in cultured enteric glia. An increasing percentage of glia responded to serotonin (4%; 100 µ M ), bradykinin (11%; 10 µ M ), and histamine (31%; 100 µ M ), whereas 100% of glia responded to ATP (100 µ M ). ATP-evoked calcium signaling was concentration dependent in terms of the percentage of glia responding and the peak [Ca 2+ ] i achieved; responses were pertussis toxin insensitive. Based on responsiveness of enteric glia to purinergic agonists and peak [Ca 2+ ] i evoked, ATP = UTP > ADP > Β,Γ-methyleneadenosine 5′-triphosphate > 2-methylthioadenosine 5′-triphosphate = Α,Β-methyleneadenosine 5′-triphosphate = AMP = adenosine, suggesting a glial P 2U receptor. Depletion of d- myo -inositol 1,4,5-trisphosphate-sensitive calcium stores by thapsigargin (10 µ M ) abolished glial responses to ATP. Similarly, calcium responses were decreased 92% by U-73122 (10 µ M ), an inhibitor of phospholipase C, and 93% by the phorbol ester phorbol 12-myristate 13-acetate (100 n M ), an activator of protein kinase C. Thus, cultured enteric glia can respond to neurotransmitters with increases in [Ca 2+ ] i . Our data suggest that glial responses to ATP are mediated by a P 2U receptor coupled to activation of phospholipase C and release of intracellular calcium stores.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66050/1/j.1471-4159.1996.66020604.x.pd

Acute gallbladder torsion - a continued pre-operative diagnostic dilemma

Acute gallbladder volvulus continues to remain a relatively uncommon process, manifesting itself usually during exploration for an acute surgical abdomen with a presumptive diagnosis of acute cholecystitis. The pathophysiology is that of mechanical organo-axial torsion along the gallbladder's longitudinal axis involving the cystic duct and cystic artery, and with a pre-requisite of local mesenteric redundancy. The demographic tendency is septua- and octo-genarians of the female sex, and its overall incidence is increasing, this being attributed to increasing life expectancy. We discuss two cases of elderly, fragile women presenting to the emergency department complaining of sudden onset right upper quadrant abdominal pain. Their subsequent evaluation suggested acute cholecystitis. Ultimately both were taken to the operating room where the correct diagnosis of gallbladder torsion was made. Pre-operative diagnosis continues to be a major challenge with only 4 cases reported in the literature diagnosed with pre-operative imaging; the remainder were found intra-operatively. Consequently, a delay in diagnosis can have devastating patient outcomes. Herein we propose a necessary high index of suspicion for gallbladder volvulus in the outlined patient demographic with symptoms and signs mimicking acute cholecystitis

Association of a genetic polymorphism (-44 C/G SNP) in the human DEFB1 gene with expression and inducibility of multiple β-defensins in gingival keratinocytes

BACKGROUND: Human β-defensins (hBDs) are antimicrobial peptides with a role in innate immune defense. Our laboratory previously showed that a single nucleotide polymorphism (SNP) in the 5' untranslated region of the hBD1 gene (DEFB1), denoted -44 (rs1800972), is correlated with protection from oral Candida. Because this SNP alters the putative mRNA structure, we hypothesized that it alters hBD1 expression. METHODS: Transfection of reporter constructs and evaluation of antimicrobial activity and mRNA expression levels in keratinocytes from multiple donors were used to evaluate the effect of this SNP on constitutive and induced levels of expression. RESULTS: Transfection of CAT reporter constructs containing the 5' untranslated region showed that the -44 G allele yielded a 2-fold increase in CAT protein compared to other common haplotypes suggesting a cis effect on transcription or translation. The constitutive hBD1 mRNA level in human oral keratinocytes was significantly greater in cells from donors with the -44 GG genotype compared to those with the common CC genotype. Surprisingly, the hBD3 mRNA level as well as antimicrobial activity of keratinocyte extracts also correlated with the -44 G allele. Induced levels of hBD1, hBD2, and hBD3 mRNA were evaluated in keratinocytes challenged with Toll-like receptor 2 and 4 ligands, interleukin-1β, TNFα, and interferon-γ (IFNγ). In contrast to constitutive expression levels, IFNγ-induced keratinocyte hBD1 and hBD3 mRNA expression was significantly greater in cells with the common CC genotype, but there was no clear correlation of genotype with hBD2 expression. CONCLUSION: The DEFB1 -44 G allele is associated with an increase in overall constitutive antimicrobial activity and expression of hBD1 and hBD3 in a manner that is consistent with protection from candidiasis, while the more common C allele is associated with IFNγ inducibility of these β-defensins and is likely to be more protective in conditions that enhance IFNγ expression such as chronic periodontitis. These results suggest a complex relationship between genetics and defensin expression that may influence periodontal health and innate immune responses

Genome-wide analysis of the human Alu Yb-lineage.

The Alu Yb-lineage is a \u27young\u27 primarily human-specific group of short interspersed element (SINE) subfamilies that have integrated throughout the human genome. In this study, we have computationally screened the draft sequence of the human genome for Alu Yb-lineage subfamily members present on autosomal chromosomes. A total of 1,733 Yb Alu subfamily members have integrated into human autosomes. The average ages of Yb-lineage subfamilies, Yb7, Yb8 and Yb9, are estimated as 4.81, 2.39 and 2.32 million years, respectively. In order to determine the contribution of the Alu Yb-lineage to human genomic diversity, 1,202 loci were analysed using polymerase chain reaction (PCR)-based assays, which amplify the genomic regions containing individual Yb-lineage subfamily members. Approximately 20 percent of the Yb-lineage Alu elements are polymorphic for insertion presence/absence in the human genome. Fewer than 0.5 percent of the Yb loci also demonstrate insertions at orthologous positions in non-human primate genomes. Genomic sequencing of these unusual loci demonstrates that each of the orthologous loci from non-human primate genomes contains older Y, Sg and Sx Alu family members that have been altered, through various mechanisms, into Yb8 sequences. These data suggest that Alu Yb-lineage subfamily members are largely restricted to the human genome. The high copy number, level of insertion polymorphism and estimated age indicate that members of the Alu Yb elements will be useful in a wide range of genetic analyses

The Iowa Homemaker vol.3, no.3-4

Table of Contents The Architectural Design of a Home by Allen Holmes Kimball, page 1 “For a Man’s House Is His Castle” by Alda Wilson, page 2 The Economics of Consumption compiled by John E. Brindley, page 3 Sunfast and Tubfast Materials by Pearl Apland, page 5 On Our Street by Juanita J. Beard, page 6 Who Is Responsible for the Child? by Orange H. Cessna, page 7 Summer Suppers by N. Beth Bailey, page 8 Vacation First Aid by Dr. Mary Sheldon, page 9 Episodes Concerning Evolution of Home Economics by Ruth Elaine Wilson, page 10 Extravagant Economics by Blanche Ingersoll, page 11 Breakfast Bridge by Eleanor Murray, page 12 Veishea Celebrates First Birthday by Helen G. Lamb, page 1

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Well Played Session: Monumental Consequence

Monumental Consequence is a single-session, live action game for classrooms, team building, and social gatherings that asks players if art is ever worth dying for. Players take on the role of villagers in the fictional town of La Ville where an army has just taken possession of their centuries old church. The villagers must come together to decide whether they risk the lives of their friends and family by attacking the church to save the precious art inside or whether they simply bomb the church and sacrifice the art to save lives. The game blends card mechanics with live action roleplaying to create a vibrant conversation about the cultural value of art and antiquities along with a seamless introduction to classroom roleplaying. This Well Played session will take participants through the entire game with an opportunity for dialogue afterward about the game’s themes, mechanics, and supporting materials

Quantifying Insolation in Multiple Shading Scenarios

This study seeks to quantify how much insolation varies over the span of a typical photovoltaic (PV) array. A solar sensor array was constructed and deployed at three locations where environmental conditions vary from full sun to highly restricted sunlight. Data was recorded at each location and then analyzed to find the effects of shading. In traditional series-parallel photovoltaic systems the total power output of the system is highly dependent on the full insolation of every cell. One cell with low insolation properties will drag down the current of an entire series string. Analysis of the sensor array data has shown that insolation varies substantially, even on an unshaded site. These findings should help, along with future research, to demonstrate the necessity of eliminating series strings from PV systems to increase energy production