Human leukocyte antigen Class II amino acid epitopes: susceptibility and progression markers for beryllium hypersensitivity

Chronic beryllium disease (CBD) is a hypersensitivity granulomatosis characterized by beryllium hypersensitivity (BH) and mediated by CD4+ T cells. However, all individuals with BH may not develop CBD. To examine the role of the three different human leukocyte antigen (HLA) Class II isotypes in BH with (CBD) and without clinical disease (BHWCD), we performed DNA-based typing of HLA-DPB1, HLA-DQB1, and HLA-DRB1 loci on 55 subjects with BH (25 with established CBD and 30 with BHWCD), and compared this with the results for 82 beryllium-exposed workers with no evidence of BH. The allele distribution was utilized to identify candidate amino acid epitopes that differed between the study groups. HLA-DPB1-E69 was the most important marker for BH, and did not differentiate BHWCD from CBD. A significant association with CBD was observed with HLA-DQB1-G86 (p(corr) < 0.04), and HLA-DRB1-S11 was significantly increased in CBD as compared with BHWCD (p < 0.03). These observations suggest that HLA-DPB1-E69 is a marker for susceptibility to BH and not just a progression marker for CBD. In addition, HLA amino acid epitopes on HLA-DRB1 and -DQB1, in concert with or independently of HLA-DPB1-E69, may be associated with progression to CBD.     

Intentional replantation of endodontically treated teeth: an update

The IR technique is a clinically successful procedure, so long as the following conditions, as outlined by Niemczyk, are met: 1) Avoid any crushing or scraping contact with the root surface or socket; 2) Root surface must be continually hydrated with tissue culture media (e.g., HBSS); 3) Tooth should be splinted, if indicated; and 4) Soft diet and hygiene instructions must be implemented and reinforced. The IR technique should not be considered a procedure of last resort. Rather, it should be used in situations where conventional apical surgery is difficult or places the patient at risk. The IR technique expands potential treatment alternatives and allows the patient to successfully retain his or her own tooth following treatment.     

Identification of pharmacokinetically stable 3, 10-dibromo-8-chlorobenzocycloheptapyridine farnesyl protein transferase inhibitors with potent enzyme and cellular activities.

Farnesyl protein transferase (FPT) is a promising target for the development of cancer chemotherapeutics because it is responsible for the farnesylation of oncogenic p21 Ras proteins which are found in nearly 30% of all human cancers and necessary for cellular development and growth. The recent discovery and progression to phase II clinical trials of trihalobenzocycloheptapyridine Sch-66336 as a potent inhibitor of FPT with oral, in vivo efficacy in mice have spawned extensive structure-activity relationship studies (SAR) of this class of compounds. Of the many trihalobenzocycloheptapyridine analogues prepared, we have identified several which inhibit FPT and cellular proliferation at single-digit nanomolar concentrations and which have good pharmacokinetic properties in mice.     

Adherence and morphology of guinea pig alveolar macrophages: effect of N-formyl methionyl peptides.

N-Formyl methionyl phenylalanine increased alveolar macrophage adherence and diameter and induced morphological changes associated with activation. N-Formyl methionyl phenylalanine may be useful in understanding macrophage activation and bacteriolytic function.     

Transgenic Chinese hamster V79 cell lines which exhibit variable levels of gpt mutagenesis

The Escherichia coli gpt gene coding for xanthine-guanine phosphoribosyl transferase has been stably transfected into HPRT- Chinese hamster V79 cells. Several gpt- cell lines have been established, which retain the sequence(s) even after long-term culture without selection for gpt. Each cell line exhibits a characteristic spontaneous mutation frequency (10(-5) to 10(-2)) in 6-thioguanine (6TG) selection. While spontaneous mutagenesis to gpt- occurs rather frequently for most cell lines, it cannot be correlated with either the number of plasmid integration sites or deletion of the plasmid sequence(s). One transgenic cell line (g12), which continuously maintains a low spontaneous mutation frequency (approximately 3 x 10(-5)), was used in comparative mutagenesis studies with wild-type V79 cells (gpt vs. hprt). Alkylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and beta-propiolactone (BPL) are shown to be equally toxic and mutagenic in both g12 and V79 cells. UV and X-rays are also equally toxic to both cell lines. The gpt locus of the g12 transfectants, however, is two to three times more sensitive to UV and 2.5-4.5 times more sensitive to X-ray mutagenesis than the endogenous hprt of wild-type V79 cells. The data presented here suggests that g12 cells may be useful to study mammalian mutagenesis by agents which yield limited response at the hprt locus. Future studies with these transgenic cells and other transgenic lines are planned to compare the mutability and repair of the same gene (gpt) at different integration sites in mammalian cells.     

Functional analysis of the effects of a fully humanized anti-CD4 antibody on resting and activated human T cells.

CD44, a cell adhesion molecule, exists in multiple isoforms that are generated by RNA alternative splicing. CD44 isoforms containing exon V6 (CD44 V6) have been associated with tumorigenesis and metastasis. We investigated the association between human B-cell activation and CD44 V6 isoform expression by analysing its expression in resting and mitogenically stimulated B cells. Results showed that resting B cells expressed the CD44 H (no variable exon) isoform alone. Activation of B cells [phorbol myristate acetate (PMA), surface immunoglobulin cross-linking alone or in the presence of interleukin-2 (IL-2)] induced CD44E (variable exon V8-10), R2 (VIO) and CD44 isoforms containing exons V6 and/or V7 (CD44 V6/V7). Epstein-Barr virus (EBV) infection of B cells, an alternative method of B-cell activation, induced the expression of CD44 E and R2 but not CD44 V6/V7. These results indicate that CD44 V6/V7 expression depends on the mode of activation. CD44 isoform expression was also investigated in a panel of EBV-negative and EBV-positive Burkitt's lymphoma (BL) B-cell lines. EBV-negative BL cells did not express CD44. In contrast, EBV-positive BL cells expressed CD44 H, R2 and E but not CD44 V6/V7 isoforms, suggesting an association between EBV infection and CD44 isoform induction. To determine directly the role of EBV in CD44 isoform induction, an EBV-negative BL cell line, BL30 (negative for all isoforms of CD44), BL30 infected in vitro with the EBNA-2-defective P3HR1 (BL30/P3HR1), and the wild-type B95-8 strain of EBV (BL30/B95-8) were examined. The parental BL30 cells infected with the wild-type EBV strain, but not with the P3HR-1 strain, expressed CD44 H, R2 and E isoforms, as seen in EBV-immortalized B cells. These studies suggest that (1) alternative splicing of CD44 isoforms is differentially regulated depending on the mode and state of cell activation, and that (2) the CD44 V6/V7 isoforms may represent B-cell activation antigens that are induced by mitogenic stimulation but not following EBV infection.     

Hemodynamic effects of elevation and stabilization of the heart during off-pump coronary surgery.

BACKGROUND: Coronary artery bypass grafting (CABG) surgery is now being performed without the use of cardiopulmonary bypass (CPB). To achieve complete myocardial revascularization off-CPB, a technique has been developed to expose target coronary arteries while hemodynamics are maintained. METHODS: Complete myocardial revascularization was performed in 18 consecutive patients. Exposure of target coronary arteries was achieved by a "single-suture" technique, placed in the oblique sinus of the pericardium. Traction on the suture elevates and rotates the heart, thereby exposing all target coronary arteries. Cardiac index (CI) and intracardiac pressures were measured with a Swan-Ganz catheter during the different phases of the operation. RESULTS: All patients were successfully operated on without CPB. There were no postoperative complications or deaths. There were no major hemodynamic changes during the different stages of the operation; in other words, CI was unchanged during elevation of the heart and snaring of the main coronary branches. Pulmonary artery wedge pressure (PAWP) increased markedly during occlusion and stabilization of the circumflex coronary artery (p < 0.05). A marked increase in CI and cardiac output (CO) from baseline values was also recorded before chest closure (p < 0.05). CONCLUSION: Complete myocardial revascularization can be achieved safely without CPB. The single-suture technique allows for exposure of all target coronary arteries without hemodynamic compromise.