Efficient expression of CFTR function with adeno-associated virus vectors that carry shortened CFTR genes

Adeno-associated virus (AAV)-based vectors have been shown to be effective in transferring the cystic fibrosis gene (CFTR) into airway epithelial cells in animal models and in patients. However, the level of CFTR gene expression has been low because the vector cannot accommodate the CFTR gene together with a promoter. In this study, we described a strategy to reduce the size of the CFTR cDNA to allow the incorporation of an effective promoter with the CFTR gene into AAV vectors. We engineered and tested 20 CFTR mini-genes containing deletions that were targeted to regions that may contain nonessential sequences. Functional analyses showed that four of the shortened CFTRs (one with combined deletions) retained the function and the characteristics of a wild-type CFTR, as measured by open probability, time voltage dependence, and regulation by cAMP. By using an AAV vector with a P5 promoter, we transduced these short forms of CFTR genes into target cells and demonstrated high levels of CFTR expression. We also demonstrated that smaller AAV/CFTR vectors with a P5 promoter expressed the CFTR gene more efficiently than larger vectors or a vector in which CFTR gene was expressed from the AAV inverted terminal repeat sequence. The CFTR mini-gene with combined deletions was packaged into AAV virions more efficiently, generated higher titers of transducing virions, and more effectively transferred CFTR function into target cells. These new vectors should circumvent the limitations of AAV vector for CFTR expression. Our strategy also may be applicable to other genes, the sizes of which exceed the packaging limit of an AAV vector.     

Nickel-titanium versus stainless-steel finger spreaders in curved canals

Fibro-osseous lesions of the sinonasal region are relatively frequent, but those strictly confined to the nasal cavity are rare. We report an atypical fibro-osseous lesion in the nasal cavity and describe its radiological features. The differential diagnosis is discussed.     

The mammalian homologue of mago nashi encodes a serum-inducible protein

OBJECTIVE: We prospectively compared the ability of two techniques--bone scintigraphy with single-photon emission computed tomography (SPECT) of the chest and CT of the chest--to reveal potential osteosarcoma metastases of the lung. SUBJECTS AND METHODS: Our study included 27 patients with osteosarcoma who prospectively underwent both bone scintigraphy with SPECT of the chest and CT of the chest. The imaging results were compared with outcome or pathologic analysis of any lung lesions found. RESULTS: Eight (30%) of the 27 patients had pulmonary metastases. Four of these eight patients had positive results on both CT studies and bone SPECT studies, with additional lesions detected with bone SPECT in two of these four patients. The other four patients with pulmonary metastases had positive results on CT studies, whereas the results of bone SPECT studies remained negative. The results of bone SPECT studies were negative in the 19 patients without pulmonary metastases. CT, however, showed abnormalities in seven (37%) of the 19 patients, which were eventually attributed to benign conditions. CONCLUSION: Negative results on a bone SPECT study of the chest should not be used to exclude the possibility of lung metastases. However, if the results are positive, a bone SPECT study can be used to confirm abnormalities seen on CT scans and may also reveal subtle lesions missed on CT scans.     

Minor histocompatibility antigens as targets for T-cell therapy after bone marrow transplantation

BACKGROUND: Topical antimicrobials have been considered for treatment of secondarily infected wounds because of the potential for reduced risk of adverse effects and greater patient convenience. We compared mupirocin cream with oral cephalexin in the treatment of wounds such as small lacerations, abrasions, or sutured wounds. METHODS: In 2 identical randomized double-blind studies, 706 patients with secondarily infected wounds (small lacerations, abrasions, or sutured wounds) received either mupirocin cream topically 3 times daily or cephalexin orally 4 times daily for 10 days. RESULTS: Clinical success at follow-up was equivalent in the two groups: 95.1% and 95.3% in the mupirocin cream and the cephalexin groups, respectively (95% confidence interval [CI], -4.0% to 3.6%; P = .89). The intention-to-treat success rate was 83% in both groups. Bacteriologic success at follow-up was also comparable: 96.9% in the mupirocin cream and 98.9% in the cephalexin groups (95% CI, -6.0% to 2.0%; P = .22). The occurrence of adverse experiences related to study treatment was similar for the 2 groups, with fewer patients in the mupirocin cream group reporting diarrhea (1.1% vs 2.3% for cephalexin). CONCLUSIONS: Mupirocin cream applied topically 3 times daily is as effective as oral cephalexin given 4 times daily for the treatment of secondarily infected wounds and was well tolerated.     

Villous papilloma of the rectum presenting as infective gastro-enteritis

Three elderly patients have recently been admitted as emergencies to the Infection Unit of the Aberdeen Hospitals as presumed cases of infective gastro-enteritis. Investigation revealed the presence of a villous papilloma of the rectum in each case.     

Hypoxic contraction of isolated canine coronary artery. Mediation by potassium-dependent exocytosis of norepinephrine

Effects of hypoxia on arterial tone, efflux of potassium, and efflux of norepinephrine were monitored for isolated canine coronary arteries labeled with radioactive potassium (42K) and norepinephrine (3HNE). Hypoxia elicited transient relaxation and subsequent sustained contraction accompanied by marked increases in the effluxes of 42K and 3HNE. After sympathetic nerve injury with 6-hydroxydopamine or cold storage, arteries responded to hypoxia with sustained relaxation. Sustained relaxation occurred also after pretreatment with L-propranolol, but not with D-propranolol or phentolamine. Inhibition of hypoxic contraction by L-propranolol did not alter 42K or 3HNE efflux. Colchicine, an inhibitor of the exocytosis of NE, suppressed hypoxic 3HNE efflux and contraction, but not 42K efflux. Proadifen inhibited 42K and 3HNE efflux as well as contraction. During proadifen-inhibited 42K efflux, exogenous K+ augmented overflow of 3HNE, indicating that proadifen relaxed the hypoxic artery primarily by inhibiting K+-dependent exocytosis of NE. The ratio of NE to dopamine beta-hydroxylase activity was similar in effluents from oxygenated arteries exposed to elevated K+ concentrations and in effluents from hypoxic arteries. Thus, hypoxia evoked exocytotic release of norepinephrine which promoted contraction by a beta-adrenergic mechanism.     

Morphology of peripheral airways in current smokers and ex-smokers

To investigate the effect of smoking status on pulmonary function and pathologic changes in the peripheral airways, we studied 97 patients who underwent thoracotomy for coin lesions. The patients were divided into 4 groups: nonsmokers (n = 9), current smokers (n = 51), and those who had ceased smoking for less than (n = 18) or more than (n = 19) 2 yr prior to surgery. We found that current smokers had evidence of air-flow obstruction with abnormal lung volumes when compared with nonsmokers. Ex-smokers had lung volumes similar to those of nonsmokers, but showed evidence of obstruction, with the FEV1/FVC between the values found for nonsmokers and current smokers. Examination of the small airways showed that the membranous bronchioles of current smokers and ex-smokers displayed only increased goblet cell metaplasia when compared with those in nonsmokers; the respiratory bronchioles of current and ex-smokers showed increases in intraluminal and airway wall inflammatory cells, wall fibrosis, and pigment deposition. We conclude that patients who currently smoke cigarettes have reduced lung function that is associated with abnormalities of airway structure. Although those who have stopped smoking have function that is closer to the nonsmoking group, there is no apparent difference in structural change between current and ex-smokers.