Glutamate receptor antagonists inhibit calpain-mediated cytoskeletal proteolysis in focal cerebral ischemia

Excitatory amino acids may promote microtubular proteolysis observed in ischemic neuronal degeneration by calcium-mediated activation of calpain, a neutral protease. We tested this hypothesis in an animal model of focal cerebral ischemia without reperfusion. Spontaneously hypertensive rats were treated with 2, 3-dihydroxy-6-nitro-7-sulfamoyl-benzo-(F)quinoxaline (NBQX), a competitive antagonist of the neuronal receptor for alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), or cis-4-[phosphono-methyl]-2-piperidine carboxylic acid (CGS 19755), a competitive antagonist of the N-methyl-d-aspartate (NMDA) receptor. After treatment, all animals were subjected to permanent occlusion of the middle cerebral artery for 6 or 24 h. Infarct volumes measured in animals pretreated with CGS 19755 after 24 h of ischemia were significantly smaller than those quantified in ischemic controls. Rats pretreated with NBQX showed partial amelioration of cytoskeletal injury with preserved immunolabeling of microtubule-associated protein 2 (MAP 2) at 6 and 24 h and reduced accumulation of calpain-cleaved spectrin byproducts only at 6 h. Prevention of cytoskeletal damage was more effective after pretreatment with CGS 19755, as shown by retention of MAP 2 immunolabeling and significant restriction of calpain activity at both 6 and 24 h. Preserved immunolabeling of tau protein was observed at 6 and 24 h only in animals pretreated with CGS 19755. Western analysis performed on ischemic cortex taken from controls or rats pretreated with either NBQX or CGS 19755 suggested that loss of tau protein immunoreactivity was caused by dephosphorylation, rather than proteolysis. These results demonstrate a crucial link between excitotoxic neurotransmission, microtubular proteolysis, and neuronal degeneration in focal cerebral ischemia.     

Recovery of muscle transfers replacing the total plantar flexor muscle group in rats

In rats, combinations of plantar flexor muscles representing approximately 20, 40, 60, and 80% of the mass of the total plantar flexor group were transferred orthotopically in the absence of synergistic muscles and allowed to recover for 120 days. We hypothesized that, compared with their individual control values for structural and functional variables, the transfers would display a hierarchical array of deficits, proportional to their initial mass and, consequently, inversely proportional to the relative load on the transfers. Surprisingly, compared with their individual control values, each muscle transfer displayed deficits of 30-40% in muscle mass, total fiber cross-sectional area, and maximum isometric force, with the exception of the smallest transfer, the plantaris (PLN) muscle, which recovered 100% of its control value for each of these variables. Therefore, except for the PLN transfer, the muscle transfers studied displayed deficits similar in magnitude to those reported for muscles transferred in the presence of synergistic muscles. The greater recovery of the PLN transfer was attributed to the relatively large requirement for force production imposed on this transfer due to the average force requirements of the total plantar flexor group.     

Acute myocardial infarction in two adolescent males

Acute myocardial infarction in previously healthy children is rare in the absence of congenital anomalies. We describe two cases of acute anterior myocardial infarction in adolescent males with no congenital heart disease, without prior history of or risk factors for coronary heart disease, and with no history of drug abuse. These cases illustrate that myocardial infarction in the absence of systemic illness or coronary anomalies can occur in an adolescent population.     

Protein-resistant coatings for glass and metal oxide surfaces derived from oligo(ethylene glycol)-terminated alkytrichlorosilanes

This paper describes the preparation of oligo(ethylene glycol)-terminated alkyltrichlorosilanes, Cl3Si(CH2)11(OCH2CH2)(n)OCH3 (n = 2, 3), and their use in the formation of self-assembled monolayers on an oxide surface. The adsorption of the trichlorosilanes from solution produces densely packed, oriented monolayer films that are 2-3 nm in thickness. The trichlorosilyl group anchors the molecules to the surface, and the resulting film exposes the ethylene glycol units at its surface, as noted by its moderate hydrophilicity (theta2(H2O) approximately 68 degrees). The films are robust with stabilities similar to those of other alkylsiloxane coatings. These oligo(ethylene glycol)-terminated silane reagents produce films that notably exhibit resistances against the non-specific adsorption of proteins from solution that are better than for films prepared from octadecyltrichlorosilane. With insulin, tysozyme, albumin, and hexokinase, no adsorption was observed with the oligo(ethylene glycol)-siloxane coatings whereas protein films of approximately a monolayer formed on surfaces-treated with octadecyltrichlorosilane. With fibrinogen, complete resistance was not possible with either coating; however, the oligo(ethylene glycol)-siloxane coatings exhibited greater resistance against non-specific adsorption. The oligo(ethylene glycol)-siloxane coatings offer performance advantages over available systems and could easily provide a direct and superior replacement in protocols that presently use silane reagents to generate hydrophobic, 'inert' surfaces.     

Molecular characterization of a common fragile site (FRA7H) on human chromosome 7 by the cloning of a simian virus 40 integration site

Common fragile sites are chromosomal loci prone to breakage and rearrangement, hypothesized to provide targets for foreign DNA integration. We cloned a simian virus 40 integration site and showed by fluorescent in situ hybridization analysis that the integration event had occurred within a common aphidicolin-induced fragile site on human chromosome 7, FRA7H. A region of 161 kb spanning FRA7H was defined and sequenced. Several regions with a potential unusual DNA structure, including high-flexibility, low-stability, and non-B-DNA-forming sequences were identified in this region. We performed a similar analysis on the published FRA3B sequence and the putative partial FRA7G, which also revealed an impressive cluster of regions with high flexibility and low stability. Thus, these unusual DNA characteristics are possibly intrinsic properties of common fragile sites that may affect their replication and condensation as well as organization, and may lead to fragility.     

The effects of space flight on the production of monorden by Humicola fuscoatra WC5157 in solid-state fermentation

BACKGROUND: There is growing evidence that laparoscopy for malignancy is associated with an increased incidence of metastasis to port sites. This study investigated the effect of different insufflation gases on port-site metastasis after laparoscopy in an established animal model. METHODS: Forty-eight Dark Agouti rats with an established adenocarcinoma in the left flank underwent laparoscopic intraperitoneal tumor laceration. The gas used for insufflation was one of the following (12 rats in each group): (1) CO2, (2) N2O, (3) helium, or (4) air. Rats were killed 7 days after the procedure, and the port sites were examined for the presence of tumor metastasis. RESULTS: Tumor involvement of port sites was significantly less likely after helium insufflation than in the other groups (p < 0.0001). There was no significant difference between the air, CO2, and N2O groups. CONCLUSIONS: This study suggests that the development of metastases in port sites after laparoscopy may be influenced in part by the choice of insufflation gas used to create the pneumoperitoneum. In particular, helium was associated with a reduced rate of metastases.     

A radiographic study of the development of the human mandibular dentition

This paper presents new data on the absolute timing of the growth of the mandibular permanent teeth, with an emphasis on the timing of the completion of the enamel crown. Two collections of cranio-dental radiographs were analyzed: (1) 267 juveniles from a contemporary pediatric dental clinic (Case Western Reserve University School of Dentistry, Cleveland, Ohio), and (2) 36 individuals sampled longitudinally from the Bolton-Brush Growth Study Center. The ages of the individuals span from three months to 18 years of age and the study includes both cross-sectional and longitudinal data. Comparisons of the ages of attainment of the Cleveland samples with other schedules of dental development are made. The relative contribution of the multiple underlying sources of variation producing the differences between the radiographically based developmental schedules remain elusive.     

The detection of gene mutation in the tubular sperm of Muta Mice following a single intraperitoneal treatment with methyl methanesulphonate or ethylnitrosourea

We have isolated a new MT-MMP related gene of 3.3 kb from a mouse lung cDNA library using a human MT1-MMP cDNA as a probe. The deduced protein sequence shows 87% homology to human MT2-MMP and 52, 50 and 29% to MT1-MMP, MT3-MMP and MT4-MMP, respectively. Thus the gene is thought to be a mouse homologue of human MT2-MMP. A monoclonal antibody raised against a synthetic peptide recognized mouse MT2-MMP as a 70 kDa protein. Like MT1- and MT3-MMPs, mouse MT2-MMP caused activation of progelatinase A upon co-transfection into COS-1 cells.     

Aspirin-like molecules that covalently inactivate cyclooxygenase-2

Many of aspirin's therapeutic effects arise from its acetylation of cyclooxygenase-2 (COX-2), whereas its antithrombotic and ulcerogenic effects result from its acetylation of COX-1. Here, aspirin-like molecules were designed that preferentially acetylate and irreversibly inactivate COX-2. The most potent of these compounds was o-(acetoxyphenyl)hept-2-ynyl sulfide (APHS). Relative to aspirin, APHS was 60 times as reactive against COX-2 and 100 times as selective for its inhibition; it also inhibited COX-2 in cultured macrophages and colon cancer cells and in the rat air pouch in vivo. Such compounds may lead to the development of aspirin-like drugs for the treatment or prevention of immunological and proliferative diseases without gastrointestinal or hematologic side effects.