AMP-activated protein kinase (AMPK) is activated in muscle of subjects with type 2 diabetes during exercise.

Insulin-stimulated GLUT4 translocation is impaired in people with type 2 diabetes. In contrast, exercise results in a normal increase in GLUT4 translocation and glucose uptake in these patients. Several groups have recently hypothesized that exercise increases glucose uptake via an insulin-independent mechanism mediated by the activation of AMP-activated protein kinase (AMPK). If this hypothesis is correct, people with type 2 diabetes should have normal AMPK activation in response to exercise. Seven subjects with type 2 diabetes and eight matched control subjects exercised on a cycle ergometer for 45 min at 70% of maximum workload. Biopsies of vastus lateralis muscle were taken before exercise, after 20 and 45 min of exercise, and at 30 min postexercise. Blood glucose concentrations decreased from 7.6 to 4.77 mmol/l with 45 min of exercise in the diabetic group and did not change in the control group. Exercise significantly increased AMPK alpha2 activity 2.7-fold over basal at 20 min in both groups and remained elevated throughout the protocol, but there was no effect of exercise on AMPK alpha1 activity. Subjects with type 2 diabetes had similar protein expression of AMPK alpha1, alpha2, and beta1 in muscle compared with control subjects. AMPK alpha2 was shown to represent approximately two-thirds of the total alpha mRNA in the muscle from both groups. In conclusion, people with type 2 diabetes have normal exercise-induced AMPK alpha2 activity and normal expression of the alpha1, alpha2 and beta1 isoforms. Pharmacological activation of AMPK may be an attractive target for the treatment of type 2 diabetes.

Phytochrome types in Picea and Pinus. Expression patterns of PHYA-Related types.

Knowledge of the genes in gymnosperms encoding the apoproteins of the plant photoreceptor phytochrome is currently scanty as for gymnosperm nuclear protein coding sequences in general. Here we report two complete cDNA-derived sequences which code for two different types of gymnosperm phytochrome. One sequence stems from Norway spruce (Picea abies) and the other from Scots pine (Pinus sylvestris). More detailed studies have shown that both types of phytochrome gene are present in Norway spruce. From phylogenetic analyses, these types appear to branch off from progenitors that are also the common ancestors of the angiosperm PHYA/PHYC and PHYB/PHYD/PHYE lineages. Partial phytochrome sequences of other gymnosperms cluster with either the one type or the other of the gymnosperm phytochrome genes characterized here. Southern blot analysis of Picea DNA using probes derived from the full-length Picea gene indicated a family of at least five members. Whether they code for new types may be doubted since only two phylogenetic clusters were found. Studies using RNA-PCR of Picea RNA extracted from either light- or dark-grown seedlings indicated that the steady-state levels of the transcripts of two PHYA/C-related genes were hardly affected by light.

T-DNA presence and opine production in tumors of Picea abies (L.) Karst induced by Agrobacterium tumefaciens A281.

The hypervirulent Agrobacterium tumefaciens strain A281 formed frequent tumors (31%) on Picea abies (Norway spruce), an economically important tree species in Swedish forests. Three-month-old seedlings were inoculated and tumors were established that grew hormone-independently in culture. Tumors contained agropine and mannopine/mannopinic acid as determined by acid pH paper electrophoresis. In addition, DNA hybridization studies showed that the DNA from these tumor lines contained sequences homologous to Ti plasmid T-DNA, whereas wild-type spruce seedling DNA did not. These results suggest that Agrobacterium vectors can be used for gene transfer into this important forest species.

Within-population variation in susceptibility to Agrobacterium tumefaciens A281 in Picea abies (L.) Karst.

The purpose of this study was to investigate the genetic variation of susceptibility to Agrobacterium within a Picea abies population. Tumor formation was studied in 16 open-pollinated families belonging to a central Swedish population of Picea abies. Strain A281 of Agrobacterium tumefaciens was used to infect 3-monthold seedlings in a five-block greenhouse experiment. The analysis of variance showed strong significance for the between-family variation of tumor-formation percentages, varying from 28% to 73%. The most susceptible material will be suitable for experiments on the production of transgenic plants in vitro using disarmed Agrobacterium strains.

Inheritance of autumn frost hardiness in Pinus sylvestris L. seedlings.

Inheritance of frost hardiness was analysed making use of a 12×12 incomplete factorial mating design. Owing to space limitations only 59 families could be tested in four experiments. To link the four experiments, some families were common to two or more experiments. The seedlings were grown in climate chambers under conditions inducing autumn hardening. The plants were exposed to a freezing temperature of -10 °C for three hours at night lengths of 11-13 h. A statistical model was developed for analyses of variance of our data. The genetic variation and the variation due to the cultivation regimes during autumn hardening were of the same magnitude. The additive effects were the most important ones for induction of frost damage. No interaction following long-distance crossing was noted. Mixed model equations were used for ranking of the parents. The results obtained support a polygenic inheritance of frost hardiness. The large within-population variation offers good opportunities for hardiness breeding.

Inheritance of bud-set and bud-flushing in Picea abies (L.) Karst.

Diallel crosses were made between clones of a French and a Swedish provenance of Picea abies. The former is characterized by a long critical night length for bud-set and a late flushing of the buds; the latter by a short critical night length for bud-set and an early flushing of the buds. The F1 hybrid seedlings and their French and Swedish intraprovenance half-sibs were tested over three growth periods in the phytotron at the College of Forestry, Stockholm. In comparison with intraprovenance half-sibs, the hybrid progenies gave, on average, an intermediate response for the photoperiodic control over budset and for the temperature requirements for bud-flushing. This indicates the prevalence of additive action of multiple factors in the determination of the photoperiodic and temperature response. However, individual interprovenance hybrid progenies revealed a range of responses, and in certain combinations the response of these hybrids and their intraprovenance half-sibs coincided. By selection of suitable parents, interprovenance crosses can be used to produce hybrids with desired photoperiodic characteristics and temperature requirements.

Reduction of myocardial reinfarction by the combined treatment with clofibrate and nicotinic acid.

In an ongoing study 558 consecutive survivors of myocardial infarction below 70 years, mean age 59 years, were randomly allocated 4 months after the acute episode into a control group or a chemotherapy group from December 1972 to April 1976. Both groups were given moderate advice about diet and the chemotherapy group was prescribed clofibrate, 1 g twice daily, and nicotinic acid 1 g three times daily. Serum cholesterol and triglycerides were lowered around 15-20% and 30% respectively in the chemotherapy group while only insignificant reductions were observed in the control group. Until December 1976 total mortality and mortality from IHD has been the same in the two groups. The number of non-fatal myocardial infarctions has been 38 in the control and 19 in the chemotherapy group, a statistically significant reduction (P less than 0.01).