The serum complement system: a simplified laboratory exercise to measure the activity of an important component of the immune system.

The immune system is a vital physiological component that affords animals protection from disease and is composed of innate and adaptive mechanisms that rely on cellular and dissolved components. The serum complement system is a series of dissolved proteins that protect against a variety of pathogens. The activity of complement in serum can be determined by its ability to lyse red blood cells in vitro.

A semester-long student-directed research project involving enzyme immunoassay: appropriate for immunology, endocrinology, or neuroscience courses.

The following project aimed at promoting integrated and long-lasting learning is described for an Immunology course, but it may be adapted to other disciplines. Students were asked to develop and carry out a research project to examine the relationship between immune function and stress. The experiments were required to include the assessment of salivary cortisol and salivary IgA (sIgA) with enzyme immunoassays.

A target-unrelated peptide in an M13 phage display library traced to an advantageous mutation in the gene II ribosome-binding site.

Screening of the commercially available Ph.D.-7 phage-displayed heptapeptide library for peptides that bind immobilized Zn2+ resulted in the repeated selection of the peptide HAIYPRH, although binding assays indicated that HAIYPRH is not a zinc-binding peptide.

NorthEast Under/graduate Research Organization for Neuroscience (NEURON): Our Third New York City Meeting.

The NorthEast Under/graduate Research Organization for Neuroscience (N.E.U.

A flexible stereospecific synthesis of polyhydroxylated pyrrolizidines from commercially available pyranosides.

Nitrogen-containing sugar analogues, known as azasugars or iminosugars, such as polyhydroxylated piperdines, pyrrolidines, pyrrolizidines, and indolizidines, have the potential to become important therapeutic agents due to their ability to inhibit glycosidases. Synthetic pathways that are able to systematically produce a variety of these azasugars are eagerly sought after, since even minute structural or stereochemical changes often significantly alter the degree of inhibition. The synthesis of tetrahydroxylated pyrrolizidines 40 and 41 starting from methyl alpha-d-glucopyranoside is described and will be used as a template to develop syntheses of all the stereoisomers of polyhydroxylated pyrrolizidine 9 as well as other analogous bicyclic polyhydroxylated iminosugars.

Haploid vegetative mycelia of Armillaria gallica show among-cell-line variation for growth and phenotypic plasticity.

Vegetative mycelial cells of Armillaria are expected to have diploid nuclei. Cells from a single mycelium therefore would not be expected to differ from one another for ecologically relevant quantitative traits. We isolated two sets of basidiome cell lines (from spores and stipe cells) and one set of vegetative cell lines (from an attached rhizomorph) from a single contiguous Armillaria gallica mycelium.

Generation of a long-lasting, protective, and neutralizing antibody response to the ranavirus FV3 by the frog Xenopus.

Xenopus serves as an experimental model to evaluate the contribution of adaptive immunity in host susceptibility to emerging ranaviral diseases that may contribute to amphibian population declines. It has been previously shown that following a secondary infection with the ranavirus frog virus 3 (FV3), adult Xenopus more rapidly clear FV3 and generate specific anti-FV3 IgY antibodies. We have further evaluated the potency and persistence of the Xenopus antibody response against FV3.

Phylogenetic conservation of gp96-mediated antigen-specific cellular immunity: new evidence from adoptive cell transfer in xenopus.

Background: In vertebrates from man to frogs, the heat shock protein (hsp) gp96 elicits T-cell responses against antigenic peptides that it chaperones. In Xenopus, immunization with gp96 purified from normal tissues accelerates rejection of MHC identical, minor histocompatibility (H) antigen-disparate skin grafts in vivo and induces MHC-restricted CTL responses in vitro. Also in Xenopus, gp96 derived from MHC class I-negative tumors elicits peptide-specific responses against these tumors in vivo and MHC-unrestricted CD8 killing in vitro.

An introductory bioinformatics exercise to reinforce gene structure and expression and analyze the relationship between gene and protein sequences.

We have developed an introductory bioinformatics exercise for sophomore biology and biochemistry students that reinforces the understanding of the structure of a gene and the principles and events involved in its expression. In addition, the activity illustrates the severe effect mutations in a gene sequence can have on the protein product. Students search GenBank for the wild-type nucleotide sequence of the Caenorhabditis elegans unc-22 gene, the amino acid sequence of its gene product, and the nucleotide sequence of the transposon Tc5.

Phenotypic plasticity and evolutionary potential in somatic cells of Armillaria gallica.

Somatic cells of Armillaria gallica fruit bodies have been shown to possess different genotypes for molecular-marker and mating-type loci. Here we report experiments on six quantitative traits and demonstrate that somatic cells of fruit bodies possess almost as much genetic variation for growth rate and phenotypic plasticity as do spores, the products of meiosis. Genetically distinct somatic cells therefore have the potential to grow at different rates relative to one another during primordial fruit body formation.

A structural representation of migraine diagnostic criteria: the experts' view.

Objective: To generate an empirically derived structural representation of migraine diagnostic criteria in a group of international headache experts using the Pathfinder network scaling algorithm in order to evaluate the validity of the migraine criteria used in the International Headache Society (IHS) classification system.

Background: Because it is a disease entity that lacks objective defining markers, developing valid diagnostic criteria for migraine is a challenge. The IHS committee relied on expert consensus to develop their classification system in 1988.

Synthesis of methyl alpha-D-glucopyranosyl-(1-->4)-alpha-D-galactopyranoside and methyl alpha-D-xylo-hex-4-ulopyranosyl-(1-->4)-alpha-D-galactopyranoside.

The syntheses of methyl alpha-D-glucopyranosyl-(1-->4)-alpha-D-galactopyranoside (1) and methyl alpha-D-xylo-hex-4-ulopyranosyl-(1-->4)-alpha-D-galactopyranoside (4) are reported. The keto-disaccharide 4 is of interest in our design, synthesis, and study of pectate lyase inhibitors. The key step in the syntheses was the high-yielding, stereospecific formation of methyl 4,6-O-benzylidene-2',3'-di-O-benzyl-alpha-D-glucopyranosyl-(1-->4)-2,3,6-tri-O-benzyl-alpha-D-galactopyranoside (15), which was accomplished by reacting 2,3-di-O-benzyl-4,6-O-benzylidene-D-glucopyranosyl trichloroacetimidate (10) with methyl 2,3,6-tri-O-benzyl-alpha-D-galactopyranoside (14) in the presence of a catalytic amount of tert-butyldimethylsilyl trifluoromethane sulfonate (TMSOTF).

A genetic mosaic in the fruiting stage of Armillaria gallica.

The basidiome stage of Armillaria gallica can be a genetic mosaic. Ten cells isolated from a single basidiome in 1986 produced nine different genotypes when analyzed for variation at six nuclear loci. Four additional basidiomes collected in 1986 produced mosaic patterns when analyzed for variation at a single nuclear (PCR-RFLP) locus.

The development of perceptual completion abilities: infants' perception of stationary, partially occluded objects.

We explored infants' ability to perceive stationary, partially occluded objects as connected units (Experiments 1 and 2) with specific appearances (Experiment 3). In each experiment, the infants saw 2 test events involving what appeared to adults to be a tall rectangular object whose middle portion was hidden behind a narrow screen. During the test events, the screen alternately uncovered and covered the object.

A relapse-prevention program: effects of electromyographic training on high and low levels of state and trait anxiety.

This study involved 40 college subjects and investigated the effects of EMG training on high and low state- and trait-anxiety scores. At pretreatment assessment subjects were administered the Spielberger State-Trait Anxiety Inventory (1970). Subjects were treated with EMG training with an established treatment criterion of 3 microvolts.