Long-term treatment of growth hormone insensitivity syndrome with IGF-I. Results of the European Multicentre Study. The Working Group on Growth Hormone Insensitivity Syndromes.

A total of 33 patients (17 female, 16 male) with Laron syndrome (n = 31) or hGH-1 gene (n = 2, type IA deletion) from 22 centres in 12 countries were enrolled in a study conducted by Pharmacia & Upjohn, Stockholm, which was designed to test the efficacy, in terms of growth promotion and safety, of IGF-I (Igef(TM)). The patients were treated with 40-120 microg/kg IGF-I s.c. twice daily after meals. After the study ended, the patients continued to be treated on an individual basis. The results of 17 patients, who were treated for 48 months or longer were available for the present analysis. Six patients were treated for up to 72 months. When treatment started, the mean age of these patients (8 female, 9 male) was 9.1 (3.7-13.5) years and mean height was -6.5 +/- 1.3 SDS. At the end of the observation period, the mean age of the 17 patients was 14.2 (9.1-17. 7) years and mean height was -4.9 +/- 1.9 SDS. All patients showed a significant increase in growth during the final year on IGF-I, with two of them reaching the age-corresponding 3rd centile. The total gain in height (DeltaHT) was 1.7 +/- 1.2 SDS. DeltaHT SDS correlated negatively with age at onset of treatment (R(2) = -0.78, p < 0.02). BMI was 0.6 +/- 1.8 SDS at start of treatment and 1.8 +/- 1.5 SDS at the end of observation. Total DeltaHT SDS correlated positively with total DeltaBMI SDS (R(2) = 0.59, p < 0.01). Long-term treatment of patients with GHIS thus proved to be effective in promoting growth. If treatment is started at an early age, there is considerable potential for achieving height normalisation. The treatment modalities need to be optimized with respect to the growth-promoting and metabolic effects of IFG-I.     

Purification and characterization of the d-xylose isomerase gene from Escherichia coli

A DNA fragment containing both the Escherichia coli d-xylose isomerase (d-xylose ketol-isomerase, EC 5.3.1.5) gene and the d-xylulokinase (ATP: d-xylulose 5-phosphotransferase, EC 2.7.1.17) gene has been cloned on an E. coli plasmid. The d-xylose isomerase gene was separated from the d-xylulokinase gene by the construction of a new deletion plasmid, pLX7. The d-xylose isomerase gene cloned on pLX7 was found still to be an intact gene. The precise location of the d-xylose isomerase gene on the plasmid pLX7 was further determined by the construction of two more plasmids, pLX8 and pLX9. This is believed to be the first d-xylose isomerase gene that has been isolated and extensively purified from any organism. d-Xylose isomerase, the enzyme product of the d-xylose isomerase gene, is responsible for the conversion of d-xylose to d-xylulose, as well as d-glucose to d-fructose. It is widely believed that yeast cannot ferment d-xylose to ethanol primarily because of the lack of d-xylose isomerase in yeast. d-Xylose isomerase (also known as d-glucose isomerase) is also used for the commercial production of high-fructose syrups. The purification of the d-xylose isomerase gene may lead to the following industrial applications: (1) cloning and expression of the gene in yeast to make the latter organism capable of directly fermenting d-xylose to ethanol, and (2) cloning of the gene on a high-copy-number plasmid in a proper host to overproduce the enzyme, which should have a profound impact on the high-fructose syrup technology.     

A simplified procedure for organic phosphorus determination from phospholipids

We describe an improved method for the determination of organic phosphorus from phospholipids. It is the combination of a very fast mineralization step followed by the estimation of liberated phosphate by means of malachite green. This method is accurate, simple, and sensitive.     

Long-term effects of deendothelialization of rabbit aorta: in vitro synthesis of DNA, protein, and lipid

To study the long-term local effects of a single balloon catheter deendothelialization of the aorta in the rabbit, the incorporation of [3H]leucine and [3H]thymidine into protein and DNA, respectively, and [14C]acetate and [14C]mevalonate into sterols was measured in incubations of intima-media sections prepared from vessels taken 1 year following the procedure. The uptake of [3H]thymidine by the tissue was essentially the same as in the nonballooned controls, but the incorporation of [3H]leucine and [14C]acetate into tissue residue (proteins and glycoproteins) was approximately nine times and four times control values, respectively. At the same time, sections from the ballooned animals incorporated over six times the amount of radioactive acetate into nonsaponifiable lipids and cholesterol than did controls. In animals ballooned 3 months before sacrifice, when about half of the aortic luminal surface was covered with endothelium, intima-media tissue from both covered and uncovered areas showed increased uptake of labeled precursors into protein, nonsaponifiables, and cholesterol but there was no significant difference in incorporation between reendothelialized and nonendothelialized areas. The persistence of increased metabolic activity in the vessel following the loss of endothelium could be a contributing factor in the atherogenic process.     

Two- and three-dimensional image reconstructions from stained and autoradiographed histological sections

A complete system has been developed to utilize histologicalserial sections for two- and three-dimensional image reconstructions.Eighty to 120 sections are digitized using a personal computingsystem augmented with a imaging board and CCD camera. The imagefiles are transmitted to a VAX computer for processing and imagereconstruction, and the processed images are transmitted backto the personal computer for display and recording using a filmrecorder or PostScript printer. The software developed for thesystem allows serial sections to be placed into proper registrationin a 256³ array, 256 grey levels. Autoradiographs of the sectionsare obtained in the presence of appropriate standards whichare used to recalibrate grey levels to represent linearly theradioactivity of each pixel in the sections and scale the valuesto allow maximum use of the grey scale. Starting from coronallysectioned material the system has been used to analyse and reconstructrat nasal turbinates. In two dimensions horizontal and sagittalsections have been obtained while in three dimensions back-to-frontand surface-rendered images have been constructed. Useful renderingof differential metabolic activity within an organ of complexgeometry has been obtained, and there appears to be no reasonwhy the system cannot be used for any material for which serialsectioning is appropriate. Received on November 29, 1989; accepted on February 28, 1990