Inhibition of glucocorticoid-induced apoptosis by targeting splice variants of \u3ci\u3eBIM\u3c/i\u3e mRNA with small interfering RNA and short hairpin RNA.

Glucocorticoids (GCs) induce apoptosis in lymphocytes and are effective agents for the treatment of leukemia. The activated glucocorticoid receptor (GR) initiates a transcriptional program leading to caspase activation and cell death, but the critical signaling intermediates in GC-induced apoptosis remain largely undefined. We have observed that GC induction of the three major protein products of the Bcl-2 relative Bim (BimEL, BimS and BimL) correlates with GC sensitivity in a panel of human pre-B acute lymphoblastic leukemia (ALL) cell lines. To test the hypothesis that Bim facilitates GC-induced apoptosis, we reduced BIM mRNA levels and Bim protein levels by RNA interference (RNAi) in highly GC-sensitive pre-B ALL cells. Reducing Bim proteins by either electroporation of synthetic siRNA duplexes or lentiviral-mediated stable expression of shRNA inhibited activation of caspase-3 and increased cell viability following GC exposure. We also observed that the extent of GC resistance correlated with siRNA silencing potency. siRNA duplexes that reduced only BimEL or BimEL and BimL (but not BimS) exhibited less GC resistance than a potent siRNA that silenced all three major isoforms, implying that induction of all three Bim proteins contributes to cell death. Finally, the modulation of GC-induced apoptosis caused by Bim silencing was independent of Bcl-2 expression levels, negating the hypothesis that the ratio of Bim to Bcl-2 regulates apoptosis. These results offer evidence that induction of Bim by GC is a required event for the complete apoptotic response in pre-B ALL cells

A Central Limit Theorem for Repeating Patterns

This note gives a central limit theorem for the length of the longest subsequence of a random permutation which follows some repeating pattern. This includes the case of any fixed pattern of ups and downs which has at least one of each, such as the alternating case considered by Stanley in [2] and Widom in [3]. In every case considered the convergence in the limit of long permutations is to normal with mean and variance linear in the length of the permutations

\u3ci\u3ePseudostertagia bullosa\u3c/i\u3e (Nematoda: Trichostrongyloidea) in Artiodactyl Hosts from North America: Redescription and Comments on Systematics

A relationship for Pseudostertagia bullosa within the trichostrongyloids has been enigmatic or unresolved. Studies of the synlophe in males and females of P. bullosa revealed a tapering system anterior to the deirids and a pattern of parallel ridges extending to near the caudal extremity in both lateral and median fields. Structurally, the synlophe differs considerably from that seen among the Cooperiinae and exhibits homoplasy with respect to ridge systems among some Ostertagiinae. Other structural characters due to symplesiomorphy, homoplasy or because they represent autapomorphies do not serve to reveal the putative relationships for P. bullosa with other trichostrongyloids. Although somewhat equivocal, the 2-2-1 pattern of the bursa and position of rays 2 and 3 suggest an association with the Cooperinae, as postulated by Durette-Desset and others. Pseudostertagia bullosa appears to be a species that has survived in the pronghorn, Antilocapra americana, a relictual pecoran artiodactyl that occurs in xeric regions of western North America; pronghorn are the sole remnant of the late Tertiary radiation for Antilocapridae across North America. Pseudostertagia bullosa may occur in mixed infections with a number or ostertagiines in the abomasa of mule deer (Odocoileus hemionus) and domestic sheep (Ovis aries) in regions of sympatry for pronghorn and these artiodactyl hosts

\u3ci\u3ePseudostertagia bullosa\u3c/i\u3e (Nematoda: Trichostrongyloidea) in Artiodactyl Hosts from North America: Redescription and Comments on Systematics

A relationship for Pseudostertagia bullosa within the trichostrongyloids has been enigmatic or unresolved. Studies of the synlophe in males and females of P. bullosa revealed a tapering system anterior to the deirids and a pattern of parallel ridges extending to near the caudal extremity in both lateral and median fields. Structurally, the synlophe differs considerably from that seen among the Cooperiinae and exhibits homoplasy with respect to ridge systems among some Ostertagiinae. Other structural characters due to symplesiomorphy, homoplasy or because they represent autapomorphies do not serve to reveal the putative relationships for P. bullosa with other trichostrongyloids. Although somewhat equivocal, the 2-2-1 pattern of the bursa and position of rays 2 and 3 suggest an association with the Cooperinae, as postulated by Durette-Desset and others. Pseudostertagia bullosa appears to be a species that has survived in the pronghorn, Antilocapra americana, a relictual pecoran artiodactyl that occurs in xeric regions of western North America; pronghorn are the sole remnant of the late Tertiary radiation for Antilocapridae across North America. Pseudostertagia bullosa may occur in mixed infections with a number or ostertagiines in the abomasa of mule deer (Odocoileus hemionus) and domestic sheep (Ovis aries) in regions of sympatry for pronghorn and these artiodactyl hosts

Evaluation of glucocorticoid sensitivity in 697 pre-B acute lymphoblastic leukemia cells after overexpression or silencing of MAP Kinase Phosphotase-1

PURPOSE: To determine the effect of reducing MAP kinase phosphatase-1 (MKP-1) levels on cell death induced by glucocorticoid (GC) or hydroxyurea (HU) treatment in the human pre-B acute lymphoblastic leukemia cell line 697. METHODS: Stable MKP-1 overexpressing transformants of the 697 pre-B ALL cell line were created and tested for sensitivity to the GC triamcinolone acetonide (TA) and HU, and compared to a control 697 cell line containing normal MKP-1 expression levels. Small interfering RNAs (siRNAs) were designed to inhibit MKP-1 expression and evaluated for their effect on GC-mediated cell death. RESULTS: MKP-1 overexpression caused a phenotype of partial resistance to HU-induced apoptosis but not to GC-induced apoptosis. Electroporation of siRNAs effectively silenced MKP-1 expression, and increased sensitivity to TA by 9.6±1.9%. CONCLUSIONS: Because MKP-1 protects certain tumor cells from chemotherapy-induced apoptosis, its inhibition is being considered as a possible strategy for combination cancer therapy. However, this study suggests that while MKP-1 inhibition may improve the efficacy of DNA damaging agents, it may have only limited utility in combination with glucocorticoids