The effects of orlistat on body weight and glycaemic control in overweight patients with type 2 diabetes: a randomized, placebo-controlled trial.

AIM: To assess the long-term effects of orlistat on body weight, glycaemic control and cardiovascular risk factors in overweight patients with type 2 diabetes. METHODS: This was a multicentre, randomized, placebo-controlled study with a 4-week placebo plus diet lead-in period and a 48-week, double-blind treatment period. Overweight or obese adults [body mass index (BMI) >or= 28 kg/m2] with HbA1c of 6.5-11% and clinical type 2 diabetes were randomized to orlistat (120 mg t.i.d. n = 189) or placebo (n = 180) in conjunction with a low-calorie diet. Patients had either received sulphonylurea therapy for at least 2 months before the study or were not receiving any antidiabetic medication (the majority of which were drug-naïve). RESULTS: After 1 year, patients in the orlistat group lost significantly more weight than patients in the placebo group (-5.4% vs. -3.6%; p = 0.006). Moreover, significantly more patients achieved weight loss of >or= 5% with orlistat compared with placebo (51.3% vs. 31.6%; p = 0.0001). Patients treated with orlistat also had significantly greater improvements than placebo-treated patients in HbA1c (-0.9% vs. -0.4%; p < 0.001), fasting glucose (-1.6 vs.-0.7 mmol/l; p = 0.004) and post-prandial glucose (-1.8 vs. -0.5 mmol/l; p = 0.003). In addition, orlistat-treated patients had a significantly greater reduction in LDL cholesterol compared with placebo. Overall, orlistat had a similar safety profile to placebo, with the exception of a higher incidence of generally mild and transient gastrointestinal events known to be associated with the mode of action of orlistat. CONCLUSIONS: Treatment with orlistat plus diet resulted in significant weight loss, improved glycaemic control and cardiovascular risk factor profile in overweight patients with type 2 diabetes.

Mutations in the hepatocyte nuclear factor-1alpha gene in MODY and early-onset NIDDM: evidence for a mutational hotspot in exon 4.

We have recently shown that mutations in the gene encoding the transcription factor hepatocyte nuclear factor (HNF)-1alpha are the cause of one form of maturity-onset diabetes of the young (MODY3). Here, we report the exon-intron organization and partial sequence of the human HNF-1alpha gene. In addition, we have screened the ten exons and flanking introns of this gene for mutations in a group of 25 unrelated white subjects from Germany who presented with NIDDM before 35 years of age and had a first-degree relative with NIDDM. Mutations were identified in nine of these individuals, suggesting that mutations in the HNF-1alpha gene are a common cause of diabetes in German subjects with early-onset NIDDM and a family history of diabetes. Thus, screening for mutations in this gene may be indicated in subjects with early-onset NIDDM. Interestingly, three of the nine mutations occurred at the same site in exon 4 with insertion of a C in a polyC tract, centered around codon 290 (designated Pro291fsinsC), thereby resulting in a frameshift during translation and premature termination. Analyses of linked DNA polymorphisms in the HNF-1alpha gene indicated that the Pro291fsinsC mutation was present on a different haplotype in each subject, implying that the polyC tract represents a mutational hot spot. We have also identified the mutation in the HNF-1alpha gene in the Jutland pedigree, one of the original MODY pedigrees reported in the literature, as being a T-->G substitution in codon 241, resulting in the replacement of a conserved Cys by Gly (C241G). The information on the sequence of the HNF-1alpha gene and its promoter region will facilitate the search for mutations in other subjects and studies of the role of the gene in determining normal beta-cell functions.

Identification and characterization of a novel hsc70-like gene in the human lung tumor cell line HS24.

We determined the cDNA sequence and analyzed the genomic structure of a novel human gene designated HS24/p52, which shows significant similarity to the ATP-binding domain of stress-70 proteins in the human lung tumor cell line HS24. The 2,203-nucleotide-long cDNA sequence is divided into an incomplete 10-nucleotide 5' nontranslated region, a 1,425-nucleotide open reading frame which codes for 474 amino acids and a 768-nucleotide 3' nontranslated region. The first 404 of the deduced 474 amino acids resemble the amino-terminal regions of Hsp70 proteins from different species. Furthermore, single amino acid and short amino acid stretches, which are thought to be essential for the ATPase mechanism and ATP-binding activity in Hsp70 proteins, are conserved in this sequence, too. The carboxy-terminal 70 amino acids exhibit no significant similarity to hsp70 nor to any other known protein sequences. The HS24/p52 gene contains at least five introns, which differ significantly from hsc70 genes with regard to their size and location within the coding sequences. The total size of this gene is more than 15 kbp. Polymerase chain reaction (PCR) experiments showed that this gene is expressed in different human cell lines and tissues and it also seems to be highly conserved between human and mouse.

[Differential therapy with oral antidiabetic drugs]. / Differentialtherapie mit oralen Antidiabetika.

The more than 3 million type II diabetics in Germany constitute a true therapeutic challenge. Type II diabetes mellitus is part of the so-called metabolic syndrome characterized by the problem of insulin resistance/hyperinsulinemia. Treatment of type II diabetes aims at reducing insulin resistance. Oral antidiabetic management must be based on diabetic diet, in conjunction--if needed--with monotherapy with acarbose or metformin. Only after exhausting these principles of management, acarbose or metformin may be combined with sulfonylurea. Primary monotherapy with insulinotropically acting sulfonylureas is, in most cases, no longer appropriate as we are learning more about the pathophysiology of metabolic syndrome.

The calcium-binding protein MRP-8 is produced by human pulmonary tumor cells.

A calcium-binding protein was isolated from serum-free culture medium of human squamous carcinoma cells (HS 24). N-Terminal sequencing of the protein yielded 30 amino acids which were identical to the N-terminus of cystic fibrosis antigen. Northern blot analysis with an oligonucleotide derived from the N-terminus resulted in the detection of a transcript of approximately 600 bases. Screening of a HS 24-cDNA library with the same oligonucleotide led to the isolation of a 381-bp-cDNA encoding a protein of 93 amino acids. The corresponding protein has been identified as the calcium-binding protein MRP-8 usually found in Macrophages.

Optical fringe reduction technique for FM laser spectroscopy.

A simple and effective fringe reduction technique for FM 200 250 laser spectroscopy is discussed. A factor of 20 fringe reduction was observed in two-tone FM spectroscopy of water vapor. Theoretical simulations are compared to the exere perimental results.

Secretion of antileucoprotease from a human lung tumor cell line.

Two human tumor cell lines were analyzed for the production of human antileucoprotease (ALP). One of them, a human squamous lung carcinoma cell line (HS-24) synthesized, as confirmed by Western blot analysis, high amounts of ALP in serum-free medium. The supernatant inhibited elastase, chymotrypsin and trypsin. Northern blot analysis with an 18-mer radiolabelled oligonucleotide, derived from an ALP specific cDNA clone, revealed a specific mRNA of about 700-800 nucleotides in HS-24 tumor cells. In contrast, a secondary human lung tumor cell line (SB-3), derived from the adrenal cortex, did not synthesize ALP when assayed under identical conditions. The supernatant inhibited only trypsin and chymotrypsin.

S-phase cell detection with a monoclonal antibody.

The use of a newly developed monoclonal antibody against Bromodeoxyuridine is demonstrated in a study with V79 Chinese hamster cells. It can replace radioactive material in cell cycle analysis studies in the way that BrdU incorporated into the DNA during S-phase is detected by immunofluorescence. By this technique the proportion of actively synthesizing S-phase cells can be determined. The proportion of such cells is a good measure with regard to the quality of a culture and/or medium in question. The method is fast and offers within 3-5 hours the possibility to analyse growth conditions in monolayers and suspension cultures.

Single-tone and two-tone AM-FM spectral calculations for tunable diode laser absorption spectroscopy.

A generalized theory for optical heterodyne spectroscopy with phase modulated laser radiation is used which allows the calculation of signal line shapes for frequency modulation spectroscopy of Lorentzian gas absorption lines. In particular, synthetic spectral line shapes for both single-tone and two-tone modulation of leadsalt diode lasers are presented in which the contributions from both amplitude and frequency modulations are included.

[Long-term therapy with human insulin. Clinical experiences]. / Langzeittherapie mit Humaninsulin. Klinische Erfahrungen.

During in-patient admission after metabolic stabilisation insulin treatment of 32 type-1-diabetics was changed from bovine insulin (Depot-Insulin Hoechst CR) to semisynthetic human insulin of the intermediary type (Depot-H-Insulin Hoechst). Out-patient follow-up over a period of 12 months showed constant satisfactory mean day blood sugar values and morning postprandial blood sugar. 14 patients, however, showed significantly increased fasting blood sugar values which could be adjusted after change of evening insulin injections to the longer acting NPH insulin without normal insulin admixture (Basal-H-insulin Hoechst). In addition, a slight but not significant reduction of daily insulin requirements could be noted. There was no change of the relation between morning and evening insulin dosage. Hypoglycaemias did not occur significantly more frequently and hypoglycaemic symptoms showed no alterations.

Continuous subcutaneous insulin infusion therapy (CSII) influences cardiovascular responses to graded exercise in patients with autonomic diabetic neuropathy of the cardiovascular system (ADNCS).

We have demonstrated previously the effect of CSII on diabetics with ADNCS. In this study 16 Type-1-diabetics with ADNCS, 20 diabetics without ADNCS and 20 non diabetic controls were examined. Mean age and mean diabetes duration were similar in all groups. Graded exercise was performed in all patients using an ergometer cycle. In patients with ADNCS resting heart rate was significantly higher. The increase in heart rate during exercise was significantly lower, maximal tolerated work load was significantly reduced, and blood pressure-heart-rate multiplication as indirect measurement of oxygen uptake was significantly reduced too. The 16 diabetics with ADNCS were divided into 2 groups: 6 patients got CSII (12 months), 10 patients continued conventional s.c. therapy. After 6 and after 12 months graded exercise was performed again. The patients on s.c. therapy showed no improvement. 3 of the 6 CSII treated patients showed a significant improvement of the above mentioned parameters. Thus, in diabetics with ADNCS, CSII might not only influence the cardiovascular reflex tests, but might also have an effect on cardiovascular responses to graded exercise.

[Combination therapy with insulin and sulfonylurea in secondary failure of sulfonylurea therapy]. / Kombinationstherapie mit Insulin und Sulfonylharnstoffen bei Sekundärversagen der Sulfonylharnstofftherapie?

Among 16 type II diabetics who were secondary failures on sulphonylurea treatment alone, eight (group A) were changed to a combined regimen of insulin and sulphonylurea (glibenclamide), the other eight (group B) received insulin alone. Patients in group A were on the combined schedule for ten days, followed by ten days of insulin alone, followed by six months of combined treatment. In the first ten days those on combined treatment in group A required on average 30% less insulin that those of group B, with a comparable metabolic state, while insulin requirements significantly rose after ten days on insulin alone. After renewed combined treatment the insulin dose could once again be reduced, but after eight weeks there was a rise in the insulin requirement, as for group B patients. During the further observation period of three months there were no significant differences in the insulin requirement between the two groups. Combination treatment with sulphonylurea and insulin in secondary failures thus, in the short term, reduces insulin requirement; but in the long term it is not significantly different from insulin treatment alone.

[Des-phe-insulin-containing intermediary insulin compared with usual commercial preparations (author's transl)]. / Des-Phe-insulinhaltiges Intermediärinsulin im Vergleich mit handelsüblichen Präparaten.

In 20 type I diabetics comparison were made between Optisulin -depot CS (25% dissolved porcine Des-Phe-insulin/insulin and 75% crystalline porcine insulin), an intermediary insulin without depot additives, and two depot-additive containing insulins (Depot Insulin Hoechst CS and insulin Leo Mixtard) during a period of hospitalization. Des-Phe-insulins are insulins in which phenylalanine has been split off from the B chain of insulin. Since mixtures of Des-Phe-insulins with crystalline insulins are stable, there is the possibility of manufacturing monospecies-intermediary insulins without depot additives. Continued surveillance of blood sugar during application of Optisulin depot CS showed good control of the diabetic metabolism when compared with the commercially available insulins. Optisulin depot CS in its action is comparable to the two commercially available depot insulins. Side effects and allergic skin changes did not occur. Thus Optisulin depot SC adds to the at present available range of Des-Phe-insulin-containing insulins.

[Are des-Phe-insulin-containing insulin combinations preferable to regular trade preparations?]. / Sind Des-Phe-Insulin-haltige Insulinmischungen handelsüblichen Präparaten vorzuziehen?

Des-phe-insulins are modified insulins in which phenylalanin is eliminated from the b-chain of the normal insulin. Preparations containing des-phe-insulin and cristalline insulin have a good stability and one can produce insulins with different profiles. 50 diabetics were treated with different des-phe-insulin containing preparations. The carbohydrate metabolism was well controlled during the application of the new preparations. There were no side effects nor skin allergies. Des-phe-insulin containing insulins present a new interesting aspect in the treatment of insulin dependent diabetes mellitus.

Efficacy of thioctic acid in the therapy of peripheral diabetic neuropathy.

The therapeutic efficacy of thioctic acid was studied in patients with peripheral diabetic neuropathy. In a double-blind study ten diabetics were treated with thioctic acid or a placebo for 21 days. In a second study ten diabetics were also treated with thioctic acid intravenously (i.v.) for 21 days. Before and on the 11th and 21st day of treatment, we examined the clinical neurological state, the vibration sense according to biothesiometry, the nerve conduction velocity, and the degree of diabetic control. In addition the patients were asked about neuropathic complaints. The therapeutic efficacy of oral or i.v. thioctic acid could not be verified by measurements of the nerve conduction velocity or the vibration sensibility. No effect of oral thioctic acid on subjective complaints was observed. However, i.v. treatment with thioctic acid resulted in a distinct improvement of subjective complaints.