Restoration of the CCAAT box or insertion of the CACCC motif activates [corrected] delta-globin gene expression

Hemoglobin A2 (HbA2), which contains delta-globin as its non-alpha-globin, represents a minor fraction of the Hb found in normal adults. It has been shown recently that HbA2 is as potent as HbF in inhibiting intracellular deoxy-HbS polymerization, and its expression is therefore relevant to sickle cell disease treatment strategies. To elucidate the mechanisms responsible for the low-level expression of the delta-globin gene in adult erythroid cells, we first compared promoter sequences and found that the delta-globin gene differs from the beta-globin gene in the absence of an erythroid Krüppel-like factor (EKLF) binding site, the alteration of the CCAAT box to CCAAC, and the presence of a GATA-1 binding site. Second, serial deletions of the human delta-globin promoter sequence fused to a luciferase (LUC) reporter gene were transfected into K562 cells. We identified both positive and negative regulatory regions in the 5' flanking sequence. Furthermore, a plasmid containing a single base pair (bp) mutation in the CCAAC box of the delta promoter, restoring the CCAAT box, caused a 5.6-fold and 2.4-fold (P < .05) increase of LUC activity in transfected K562 cells and MEL cells, respectively, in comparison to the wild-type delta promoter. A set of substitutions that create an EKLF binding site centered at -85 bp increased the expression by 26.8-fold and 6.5-fold (P < .05) in K562 and MEL cells, respectively. These results clearly demonstrate that the restoration of either an EKLF binding site or the CCAAT box can increase delta-globin gene expression, with potential future clinical benefit.     

Morphology of early retinal lesions after experimental venous occlusion

PURPOSE: Study of early hystological lesions after experimental branch venous occlusion (BVO) in miniature pigs. MATERIALS AND METHODS: The retina was taken 1, 2, 4, 6 and 8 hours after BVO with green argon laser for examination with conventional and electronic microscopy. RESULTS: 1 hour after occlusion an extracellular focal oedema is observable in the ganglion cells optic nerve fibers and cells of inner plexiform layer; 2 hours after occlusion there is also an extracellular oedema of the external plexiform layer as intracellular oedema of the inner nuclear layer, 4 hours after occlusion the extracellular oedema is more diffuse especially in the ganglion cells, inner nuclear and optic nerve fibers layers. Cytoplasmic vacuolisation, nuclear pycnosis and ruptured cell membrane layers are observed; 8 hours after occlusion both intra and extracellular oedema are observable throughout the inner retina. CONCLUSIONS: After BVO histological lesions are found within one hour only following the occlusion, apoptosis and cell necrosis are present as soon as 4 hours after experimental BVO.     

Ultrastructure, immunofluorescence, western blot, and PCR analysis of eight isolates of Encephalitozoon (Septata) intestinalis established in culture from sputum and urine samples and duodenal aspirates of five patients with AIDS

ONO-5046 is a potent, specific and intravenously active inhibitor of neutrophil elastase. To examine the role of elastase in glomerulonephritis, we tested the effects of ONO-5046 on nephrotoxic serum (NTS) nephritis in a rat model of the disease in humans. Rats were administered ONO-5046 or phosphate-buffered saline (PBS) intraperitoneally 24 hours prior to injection of NTS, and they were then given equal doses of ONO-5046 or PBS three hours and 1, 2, 3, 4, 5 and 6 days later. Compared with the control groups, ONO-5046 significantly reduced proteinuria and hematuria, and suppressed the formation of crescentic glomeruli in a dose-dependent manner. Our results suggest that neutrophil elastase participates in NTS nephritis by degrading glomerular basement membrane proteins, and that the elastase inhibitor, ONO-5046, suppresses crescentic formation and glomerular injury caused by elastase.     

Biologically erodable microspheres as potential oral drug delivery systems

Biologically adhesive delivery systems offer important advantages over conventional drug delivery systems. Here we show that engineered polymer microspheres made of biologically erodable polymers, which display strong adhesive interactions with gastrointestinal mucus and cellular linings, can traverse both the mucosal absorptive epithelium and the follicle-associated epithelium covering the lymphoid tissue of Peyer's patches. The polymers maintain contact with intestinal epithelium for extended periods of time and actually penetrate it, through and between cells. Thus, once loaded with compounds of pharmacological interest, the microspheres could be developed as delivery systems to transfer biologically active molecules to the circulation. We show that these microspheres increase the absorption of three model substances of widely different molecular size: dicumarol, insulin and plasmid DNA.     

Second free flaps in head and neck reconstruction

Over the past decade, free-tissue transfer has greatly improved the quality of oncology-related head and neck reconstruction. As this technique has developed, second free flaps have been performed for aesthetic improvement of the reconstructed site. This study evaluated the indications for and the success of second free flaps. Medical files for patients who underwent second free flaps for head and neck reconstruction at the University of Texas M.D. Anderson Cancer Center, from May 1, 1988 to November 30, 1996, were reviewed. The flaps were classified as being either immediate (done within 72 hr) or delayed (done within 2 years) reconstructions. Indications, risk factors, recipient vessels, outcome, and complications were analyzed. Of the 28 patients included in this study, 12 had immediate (nine as salvage after primary free flap failure, and three for reconstruction of a soft-tissue defect), and 16 had delayed second free flaps (two for reconstruction of a defect resulting from excision of recurrent tumors, and 14 for aesthetic improvement). Reconstruction sites included the oral cavity in 18 patients; the midface in six; the skull base in two; and the scalp in two. The success rate for the second free flaps was 96 percent. Five patients had significant wound complications. In a substantial number of cases, identical recipient vessels were used for both the first and second free flaps. The authors conclude that second free flaps can play an important role in salvaging or improving head and neck reconstruction in selected patients. In many cases, the same recipient vessels can be used for both the first and second flaps.     

Hazardous materials incidents: the Washington Poison Center experience and approach to exposure assessment

Hazardous materials incidents include gas and vapor releases, spills, explosions, and fires. Incidents involving human exposure are challenging for most health care providers because of the vast number of potential chemicals involved, frequently incomplete incident information, and limited experience in exposure assessment. To facilitate improved evaluation and treatment of patients with chemical exposures, the Washington Poison Center established the Hazardous Materials Exposure Information Service in 1994. During the first 33 months of operation, this service has provided information on 70 incidents, involving a total of 1120 exposed individuals, including 501 patients treated in medical facilities. This paper reviews these incidents, the process used to collect information from the incidental scene, and selected techniques for evaluating the extent of individual chemical exposure.     

Genetic polymorphism of CCR5 gene and HIV disease: the heterozygous (CCR5/delta ccr5) genotype is neither essential nor sufficient for protection against disease progression. Swiss HIV Cohort

Homozygous (delta ccr5/delta ccr5) and heterozygous (CCR5/delta ccr5) deletions in the beta-chemokine receptor 5 (CCR5) gene, which encodes for the major co-receptor for macrophage-tropic HIV-1 entry, have been implicated in resistance to HIV infection and in protection against disease progression, respectively. The CCR5/delta ccr5 genotype was found more frequently in long-term nonprogressors (LTNP) (31.0%) than in progressors (10.6%, p < 0.0001), in agreement with previous studies. Kaplan-Meier survival analyses showed that a slower progression of disease, i.e. higher proportion of subjects with CD4+ T cell counts > 500/microl (p = 0.0006) and a trend toward a slower progression to AIDS (p = 0.077), was associated with the CCR5/delta ccr5 genotype. However, when LTNP were analyzed separately, no significant differences in CD4+ T cell counts (p = 0.12) and viremia levels (p = 0.65) were observed between the wild-type (69% of LTNP) and the heterozygous (31.0%) genotypes. Therefore, there are other factors which play a major role in determining the status of nonprogression in the majority of LTNP. Furthermore, there was no evidence that the CCR5/delta ccr5 genotype was associated with different rates of disease progression in the group of progressors. Taken together, these results indicate that the CCR5/delta ccr5 genotype is neither essential nor sufficient for protection against the progression of HIV disease.     

Pancreatitis and agenesis of the dorsal pancreas

We report a case of agenesis of the dorsal pancreas, complicated by pancreatitis and diabetes mellitus. A 39-year-old woman was referred for evaluation of a chronic pancreatitis. Abdominal spiral CT and ERP and MRCP demonstrated agenesis of the dorsal pancreas. The pathogenesis, clinical features and diagnosis of this very rarely reported disease are discussed.     

Identification of rheological and structural characteristics of foamable poly(ethylene terephthalate) by reactive extrusion

The reactivity and efficiency of five low molecular weight multifunctional anhydride and epoxy compounds as chemical modifiers of a bottle grade poly(ethylene terephthalate) (PET) resin were evaluated by reactive extrusion under controlled conditions. The two dianhydrides and the three epoxy compounds were used at concentrations based on stoichiometry derived from the measured carboxyl and hydroxyl end group contents of the base resin. Measures of melt viscosity, melt strength, intrinsic viscosity and carboxyl group content were used as criteria of the extent of the modification. Correlations of die pressure with extrudate swell during extrusion, and melt flow index (MFI) with melt strength by off‐line testing of the extrudates permitted the ranking of the modifiers according to their chain‐extending/branching efficiency. For some systems molecular weight increases (related to die pressure/MFI/intrinsic viscosity) accompanied by broadening of the molecular weight distribution (related to die swell/melt strength) were considered excessive. Extrusion foaming experiments with one particular dianhydride modifier that increased the intrinsic viscosity of the resin from 0.71 to 0.9 dl g^?1 indicate that production of low‐density foams by a process involving one‐step reactive modification/gas injection foaming is feasible, at conditions not significantly different from those employed in the simple reactive modification of the PET resin. The rheological and structural parameters determined in this work may be used as criteria to specify PET foamable compositions in terms of types and concentrations of modifiers. Copyright © 2004 Society of Chemical Industry