We studied 24-h profiles of circulating leptin levels using a sensitive and specific RIA in lean controls and obese subjects with or without non-insulin-dependent diabetes mellitus (NIDDM) during normal routine activity. Serum leptin levels were significantly higher in obese (41.7 Ϯ 9.0 ng/ml; n ϭ 11) and obese NIDDM (30.8 Ϯ 6.7; n ϭ 9) subjects compared with those in lean controls (12.0 Ϯ 4.4, n ϭ 6). In all the three groups, serum leptin levels were highest between midnight and early morning hours and lowest around noon to midafternoon. The nocturnal rise in leptin levels was significant when data were analyzed by ANOVA (lean: F ϭ 3.17, P Ͻ 0.0001, n ϭ 4; obese: F ϭ 2.02, P Ͻ 0.005, n ϭ 11; and obese NIDDM: F ϭ 4.9, P Ͻ 0.0001, n ϭ 5). The average circadian amplitude between acrophase and nadir was 75.6% in lean, 51.7%, in obese and 60.7% in obese NIDDM groups, respectively. No significant correlations ( P Ͼ 0.05) were observed between circulating levels of leptin and either insulin or glucose levels in any of the 20 subjects studied for 24-h profiles. The nocturnal rise in leptin observed in the present study resembles those reported for prolactin, thyroid-stimulating hormone, and free fatty acids. We speculate that the nocturnal rise in leptin could have an effect in suppressing appetite during the night while sleeping. ( J.

Vectors based on human adenovirus (Ad) and adenovirus (NAB). Approximately two of three patients demonassociated virus (AAV) are being evaluated for human strated CD4 + T cells that proliferated to Ad antigens of gene therapy. The response of the host to the vector, in which most were of the TH 1 subset, based on cytokine terms of antigen-specific immunity, will play a substantial secretion. A substantially different pattern of immune role in clinical outcome. We have surveyed cohorts of norresponses was observed to AAV2. Although virtually all mal subjects and cystic fibrosis patients for pre-existing patients had Ig to AAV2, most of these antibodies were immunity to these viruses, caused by naturally acquired not neutralizing (32% NAB) and only 5% of patients had infections. A number of humoral and cellular assays to peripheral blood lymphocytes that proliferated in response adenovirus serotype 5 (Ad5) and adeno-associated virus to AAV2 antigens. These studies demonstrate marked hetserotype 2 (AAV2) were performed from serum and peripherogeneity in pre-existing immunity to Ad5 and AAV2 in eral blood mononuclear cells. Virtually all subjects had Ig human populations. The impact of these findings on outto Ad5 although only 55% of these antibodies neutralized come following gene therapy will require further study.

Obese (ob) gene expression in abdominal subcutaneous adipocytes from lean and obese humans was examined. The full coding region of the ob gene was isolated from a human adipocyte cDNA library. Translation of the insert confirmed the reported amino acid sequence. There was no difference in the sequence of an reverse transcription PCR product of the coding region from five lean and five obese subjects. The nonsense mutation in the ob mouse which results in the conversion of arginine 105 to a stop codon was not present in human obesity. In all 10 human cDNAs, arginine 105 was encoded by CGG, consequently two nucleotide substitutions would be required to result in a stop codon. To compare the amount of ob gene expression in lean and obese individuals, radiolabed primer was used in the PCR reaction with (Iactin as a control. There was 72% more ob gene expression (P < 0.01) in eight obese subjects (body mass index, BMI = 42.8±2.7) compared to eight lean controls (BMI = 22.4±0.8). Regression analysis indicated a positive correlation between BMI and the amount of ob message (P < 0.005). There was no difference in the amount of ,B-actin expression in the two groups. These results provide evidence that ob gene expression is increased in human obesity; furthermore, the mutations present in the mouse ob gene were not detected in the human mRNA population. (J. Clin. Invest. 1995. 95:2986-2988

Ornithine transcarbamylase deficiency (OTCD) is an inborn error of urea synthesis that has been considered as a model for liver-directed gene therapy. Current treatment has failed to avert a high mortality or morbidity from hyperammonemic coma. Restoration of enzyme activity in the liver should suffice to normalize metabolism. An E1- and E4-deleted vector based on adenovirus type 5 and containing human OTC cDNA was infused into the right hepatic artery in adults with partial OTCD. Six cohorts of three or four subjects received 1/2 log-increasing doses of vector from 2 x 10(9) to 6 x 10(11) particles/kg. This paper describes the experience in all but the last subject, who experienced lethal complications. Adverse effects included a flu-like episode and a transient rise in temperature, hepatic transaminases, thrombocytopenia, and hypophosphatemia. Humoral responses to the vector were seen in all research subjects and a proliferative cellular response to the vector developed in apparently naive subjects. In situ hybridization studies showed transgene expression in hepatocytes of 7 of 17 subjects. Three of 11 subjects with symptoms related to OTCD showed modest increases in urea cycle metabolic activity that were not statistically significant. The low levels of gene transfer detected in this trial suggest that at the doses tested, significant metabolic correction did not occur.