Cystoopsis atractostei n. sp. (Nematoda: Cystoopsidae) from the subcutaneous tissue of the tropical gar,Atractosteus tropicus (Pisces), in Mexico

A new species of parasitic nematode, Cystoopsis atractostei (Trichinelloidea: Cystoopsidae), is described based on female specimens recovered from the subcutaneous tissue of the tropical gar, Atractosteus tropicus Gill (Lepisosteiformes: Lepisosteidae), from 2 localities (Canal Nueva Esperanza and Canal Tabasquillo) of the Pantanos de Centla Biological Reserve, State of Tabasco, southeastern Mexico, collected in April 2001. The total prevalence was 13%, and the mean intensity of infection was 1 nematode per fish. The new species differs from females of the only other adequately described congeneric species, C. acipenseris Wagner, 1867, mainly in possessing a bulbous inflation at the anterior end of the muscular esophagus, the vulva situated well posterior to the nerve ring, smooth cuticle, and in the shape of the posterior vesicular portion of the body (markedly transversely oval) in the largest specimens. Both species also differ in their host types (Lepisosteiformes vs. Acipenseriformes) and in geographical distribution (tropical southern Mexico vs. temperate zones of the Holarctic).     

Poly (ADP-ribose) polymerase cleavage monitored in situ in apoptotic cells

During apoptosis, the activation of a family of cysteine proteases, or caspases, results in proteolytic cleavage of numerous substrates. Antibody probes specific for neoepitopes on protein fragments generated by caspase cleavage provide a means to monitor caspase activity at the level of the individual cell. Poly (ADP-ribose) polymerase (PARP), a nuclear enzyme involved in DNA repair, is a well-known substrate for caspase-3 cleavage during apoptosis. Its cleavage is considered to be a hallmark of apoptosis. Here, we demonstrate that an affinity-purified polyclonal antibody to the p85 fragment of PARP is specific for apoptotic cells. Western blots show that the antibody recognizes the 85-kDa (p85) fragment of PARP but not full-length PARP. We demonstrate a time course of PARP cleavage and DNA fragmentation in situ using the PARP p85 fragment antibody and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) in Jurkat cells treated with anti-Fas. Furthermore, our results indicate that the p85 fragment of PARP resulting from caspase cleavage during apoptosis is rapidly localized outside the condensed chromatin but not in the cytoplasm.     

A novel class II-binding motif selects peptides that mediate organ-specific autoimmune disease in SWXJ,SJL/J,and SWR/J mice

Idiopathic dilated cardiomyopathy (DCM) is responsible for approximately 25% of all cases of congestive heart failure. We have recently shown that immunization of autoimmune-susceptible SWXJ mice with whole cardiac myosin leads to T cell-mediated experimental autoimmune myocarditis (EAMC) and DCM. We have now identified two disease-inducing peptides from cardiac alpha-myosin heavy chain (CAMHC). Our approach involved the use of a novel MHC class II-binding motif contained in several peptides known to be immunogenic in SWXJ (H-2(q,s)) mice or in the parental SJL/J (H-2(s)) or SWR/J (H-2(q)) mouse strains. Two of four CAMHC peptides containing the -KXXS- peptide motif were found to be immunogenic. Immunization of SWXJ or parental SJL/J and SWR/J mice with CAMHC peptides palpha406-425 or palpha1631-1650 resulted in EAMC and DCM, characterized by inflammation, fibrosis, and decompensated right-sided ventricular dilatation. Despite mediating high incidences of severe disease, both peptides were found to be cryptic determinants, thereby providing further evidence for the importance and perhaps predominance of self crypticity in autoimmunity. Both peptides showed dual parental I-A(q) and I-A(s) restriction and mediated passive transfer of disease with activated CD4(+) T cells. An intact motif was necessary for antigenicity because loss of activity occurred in peptides containing nonconservative substitutions at the motif's terminal lysine and serine residues. Our studies provide a new model for EAMC and DCM in strains of mice widely used in autoimmune studies. Moreover, the -KXXS- motif may be particularly useful in implicating previously overlooked proteins as autoimmune targets and in facilitating the development of new organ-specific autoimmune mouse models for human diseases.     

Procamallanus (Spirocamallanus) chetumalensis n. sp. (Nematoda: Camallanidae) from the Mayan sea catfish,Ariopsis assimilis,off the Caribbean coast of México

Procamallanus (Spirocamallanus) chetumalensis n. sp. is described from the stomach and intestine of the Mayan sea catfish Ariopsis assimilis (Günther, 1864), from the Bay of Chetumal, Quintana Roo, México. It is characterized by bifurcate deirids; males have 3 pairs of preanal papillae, 6 pairs of postanal papillae, 2 pairs of transverse elongate adanal papillae surrounding the cloacal aperture, wide caudal alae, spicules of unequal length, and a gubernaculum, and females have a rounded tail bearing a digit-like process terminating in 2 spines. This is the seventh Procamallanus (Spirocamallanus) species reported from fishes in Mexico and the first one recorded in sea catfishes of the Ariidae.     

Excess copper predisposes photosystem II to photoinhibition in vivo by outcompeting iron and causing decrease in leaf chlorophyll

Photoinhibition of photosystem II was studied in vivo with bean (Phaseolus vulgaris) plants grown in the presence of 0.3 (control), 4, or 15 microM Cu(2+). Although photoinhibition, measured in the presence of lincomycin to block concurrent recovery, is faster in leaves of Cu(2+)-treated plants than in control leaves, thylakoids isolated from Cu-treated plants did not show high sensitivity to photoinhibition. Direct effects of excess Cu(2+) on chloroplast metabolism are actually unlikely, because the Cu concentration of chloroplasts of Cu-treated plants was lower than that of their leaves. Excess Cu in the growth medium did not cause severe oxidative stress, collapse of antioxidative defenses, or loss of photoprotection. Thus, these hypothetical effects can be eliminated as causes for Cu-enhanced photoinhibition in intact leaves. However, Cu treatment lowered the leaf chlorophyll (Chl) concentration and reduced the thylakoid membrane network. The loss of Chl and sensitivity to photoinhibition could be overcome by adding excess Fe together with excess Cu to the growth medium. The addition of Fe lowered the Cu(2+) concentration of the leaves, suggesting that Cu outcompetes Fe in Fe uptake. We suggest that the reduction of leaf Chl concentration, caused by the Cu-induced iron deficiency, causes the high photosensitivity of photosystem II in Cu(2+)-treated plants. A causal relationship between the susceptibility to photoinhibition and the leaf optical density was established in several plant species. Plant species adapted to high-light habitats apparently benefit from thick leaves because the rate of photoinhibition is directly proportional to light intensity, but photosynthesis becomes saturated by moderate light.     

Activation of pyk2/related focal adhesion tyrosine kinase and focal adhesion kinase in cardiac remodeling

Cellular remodeling during progression of dilation involves focal adhesion contact reorganization. However, the signaling mechanisms and structural consequences leading to impaired cardiomyocyte adhesion are poorly defined. These events were studied in tropomodulin-overexpressing transgenic mice that develop dilated cardiomyopathy associated with chronic elevation of intracellular calcium. Analysis of tropomodulin-overexpressing transgenic hearts by immunoblot and confocal microscopy revealed activation and redistribution of signaling molecules known to regulate adhesion. Calcium-dependent pyk2/related focal adhesion tyrosine kinase (RAFTK) showed changes in expression and phosphorylation state, similar to changes observed for a related downstream target molecule of pyk2/RAFTK termed focal adhesion kinase. Paxillin, the target substrate molecule for focal adhesion kinase phosphorylation, was redistributed in tropomodulin-overexpressing transgenic hearts with enhanced paxillin phosphorylation and cleavage. Certain aspects of the in vivo signaling phenotype including increased paxillin phosphorylation could be recapitulated in vitro using neonatal rat cardiomyocytes infected with recombinant adenovirus to overexpress tropomodulin. In addition, increasing intracellular calcium levels with ionomycin induced pyk2/RAFTK phosphorylation, and adenovirally mediated expression of wild-type pyk2/RAFTK resulted in increased phospho-pyk2/RAFTK levels and concomitant paxillin phosphorylation. Collectively, these results delineate a cardiomyocyte signaling pathway associated with dilation that has potential relevance for cardiac remodeling, focal adhesion reorganization, and loss of contractility.     

Stereoselective pharmacokinetics and metabolism of flobufen in guinea pigs

Stereoselective aspects of pharmacokinetics and metabolism of a chiral nonsteroidal antiinflammatory drug, flobufen, 4-(2', 4'-difluorobiphenyl-4-yl)-2-methyl-4-oxobutanoic acid, were studied in male guinea pigs after p.o. administration of racemic flobufen (rac-flobufen) at a dose of 10 mg/kg. Blood samples were collected at intervals over 16 h after the administration of rac-flobufen for the quantification of flobufen enantiomers and their respective metabolites in plasma by chiral high-performance liquid chromatography (HPLC). Compartmental pharmacokinetic analysis was used to determine pharmacokinetic parameters of R- and S-flobufen. The plasma concentrations of the S- and R-enantiomers differed significantly during the experimental period. The S/R-enantiomeric ratio in 7plasma reached a maximum value of 10.1 at 240 min postdose. The oral clearance value of R-flobufen was five times higher than S-flobufen. The other pharmacokinetic parameters (K(e), T(1/2), V(SS)/F, MRT) of the enantiomers also differed substantially. All four stereoisomers of the dihydrometabolite of flobufen were detected in plasma with varying concentrations. Metabolite 17203 [4-(2,4-difluorophenyl)-phenylacetic acid] exhibited a relatively longer residence time compared to that noted for the enantiomers of the parent compound. Pharmacokinetics of the flobufen enantiomers were stereoselective in guinea pigs. The metabolism of flobufen was complex. However, metabolite 17203 seemed to be the main metabolite of flobufen that may be responsible for its relatively long-lasting antiphlogistic and immunomodulatory effects.     

2,6,8,9-tetrasubstituted purines as new CDK1 inhibitors

Purine inhibitors of cyclin-dependent kinases attract attention as potential anticancer drugs because their first representative roscovitine recently entered clinical trials. Although well described in terms of structure-activity relationships, we still present here a novel modification of the purine scaffold influencing their inhibitory properties. The introduced C-8 substituents, however, lowered the CDK inhibitory activity of roscovitine, whereas the antiproliferative potential of several derivatives remained high.     

Three new Procamallanus (Spirocamallanus) species from freshwater fishes in Mexico

The following 3 new species of Procamallanus (Spirocamallanus) are described from the intestines of freshwater fishes in Mexico, all belonging to the morphological group characterized by the presence of wide caudal alae, 3 pairs of subventral preanal papillae, and unequal spicules in the male: Procamallanus (Spirocamallanus) jaliscensis n. sp. (type host: Agonostomus monticola) and Procamallanus (Spirocamallanus) gobiomori n. sp. (hosts: Gobiomorus maculatus [type host], Gobiomorus polylepis and Eleotris picta) from 2 rivers in Jalisco State, western Mexico, and Procamallanus (Spirocamallanus) mexicanus n. sp. (type host: Cichlasoma geddesi) from Xalapa District, Veracruz State (Gulf of Mexico region), southeastern Mexico. Procamallanus jaliscensis is characterized by the length of the spicules (606-900 microm and 282-354 microm), number (15-16) of spiral ridges in the buccal capsule, and the digit-like protrusion with 1 terminal cuticular spike on the female tail; P. mexicanus by the length of the spicules (456-480 microm and 231-233 microm), number (10-12) of spiral ridges in the capsule, and the shape of the female tail (conical with a suddenly narrowed distal part, without any terminal spikes); and P. gobiomori by the length of spicules (318-348 microm and 156-192 microm), number (8-10) of spiral ridges and by the digit-like protrusion with 2 terminal cuticular spikes on the female tail.