Similar carbohydrate but enhanced lactate utilization during exercise after 9 wk of acclimatization to 5,620 m

We hypothesized that reliance on lactate as a means of energy distribution is higher after a prolonged period of acclimatization (9 wk) than it is at sea level due to a higher lactate Ra and disposal from active skeletal muscle. To evaluate this hypothesis, six Danish lowlanders (25 +/- 2 yr) were studied at rest and during 20 min of bicycle exercise at 146 W at sea level (SL) and after 9 wk of acclimatization to 5,260 m (Alt). Whole body glucose Ra was similar at SL and Alt at rest and during exercise. Lactate Ra was also similar for the two conditions at rest; however, during exercise, lactate Ra was substantially lower at SL (65 micro mol. min(-1). kg body wt(-1)) than it was at Alt (150 micro mol. min(-1). kg body wt(-1)) at the same exercise intensity. During exercise, net lactate release was approximately 6-fold at Alt compared with SL, and related to this, tracer-calculated leg lactate uptake and release were both 3- or 4-fold higher at Alt compared with SL. The contribution of the two legs to glucose disposal was similar at SL and Alt; however, the contribution of the two legs to lactate Ra was significantly lower at rest and during exercise at SL (27 and 81%) than it was at Alt (45 and 123%). In conclusion, at rest and during exercise at the same absolute workload, CHO and blood glucose utilization were similar at SL and at Alt. Leg net lactate release was severalfold higher, and the contribution of leg lactate release to whole body lactate Ra was higher at Alt compared with SL. During exercise, the relative contribution of lactate oxidation to whole body CHO oxidation was substantially higher at Alt compared with SL as a result of increased uptake and subsequent oxidation of lactate by the active skeletal muscles.     

Differentiation and growth of kype skeletal tissues in anadromous male Atlantic salmon (Salmo salar)

The re-initiation of bone development in adult starving Atlantic salmon (Salmo salar) during their energetically expensive upstream migration is remarkable and deserves closer examination. Dramatic alterations of the skull bones and teeth, most prominently, the development of a kype in males, are widely known but little studied or understood. We describe the microstructure and the cellular processes involved in the formation of the skeletal tissues of the kype. Fresh bone material, obtained from animals migrating upstream was subjected to radiological, histological or histochemical analysis. We show that the kype is, in part, composed of rapidly growing skeletal needles arising at the tip of the dentary. Proximally, the needles anastomose into a spongiosa-like meshwork which retains connective tissue inside bone marrow spaces. Ventrally, the needles blend into Sharpey fiber bone. Skeletal needles and Sharpey fiber bone can be distinguished from the compact bone of the dentary by radiography. Rapid formation of the skeleton of the kype is demonstrated by the presence of numerous osteoblasts, a broad distal osteoid zone, and the appearance of proteoglycans at the growth zone. The mode of bone formation in anadromous males can be described as 'making bone as fast as possible and with as little material as possible'. Unlike the normal compact bone of the dentary, the new skeletal tissue contains chondrocytes and cartilaginous extracellular matrix. Formation of the skeleton of the kype resembles antler development in deer (a form of regeneration), or hyperostotic bone formation in other teleost fishes, rather than periosteal bone growth. The type of boneformation may be understandable in the light of the animals' starvation and the energetic costs of upstream migration. However, the structured and regulated mode of bone formation suggests that the skeleton of the kype has functional relevance and is not a by-product of hormonal alterations or change of habitat.     

Targeting glioblastoma multiforme with an IL-13/diphtheria toxin fusion protein in vitro and in vivo in nude mice

Fusion proteins composed of tumor binding agents and potent catalytic toxins show promise for intracranial therapy of brain cancer and an advantage over systemic therapy. Glioblastoma multiforme (GBM) is the most common form of brain cancer and overexpresses IL-13R. Thus, we developed an interleukin-13 receptor targeting fusion protein, DT(390)IL13, composed of human interleukin-13 and the first 389 amino acids of diphtheria toxin. To measure its ability to inhibit GBM, DT(390)IL13 was tested in vitro and found to inhibit selectively the U373 MG GBM cell line with an IC(50) around 12 pmol/l. Cytotoxicity was neutralized by anti-human-interleukin-13 antibody, but not by control antibodies. In vivo, small U373 MG glioblastoma xenografts in nude mice completely regressed in most animals after five intratumoral injections of 1 microg of DT(390)IL13 q.o.d., but not by the control fusion protein DT(390)IL-2. DT(390)IL13 was also tested against primary explant GBM cells of a patient's excised tumor and the IC(50) was similar to that measured for U373 MG. Further studies showed a therapeutic window for DT(390)IL13 of 1-30 microg/injection and histology studies and enzyme measurements showed that the maximum tolerated dose of DT(390)IL13 had little effect on kidney, liver, spleen, lung and heart in non-tumor-bearing immunocompetent mice. Together, these data suggest that DT(390)IL13 may provide an important, alternative therapy for brain cancer.     

Identification and characterization of two novel low-molecular-weight dual specificity phosphatases

We have cloned and characterized two novel human low molecular weight dual specificity phosphatases (LMW-DSPs). Both genes are expressed exclusively in the testis, but are not altered in any of several disease states examined. Transfection into COS cells indicates that both proteins are expressed in the nucleus and the cytoplasm. Both proteins are able to dephosphorylate the phosphotyrosine analog pNPP in vitro and can be inhibited by sodium orthovanadate. In vitro experiments also demonstrate that both DSPs can dephosphorylate single and diphosphorylated synthetic MAPK peptides, with preference for the phosphotyrosine and diphosphorylated forms over phosphothreonine. However, when co-transfected with MAPKs into COS cells, the novel DSPs exhibited no detectable in vivo activity against MAPKs under our conditions. Our data suggest that these novel LMW-DSPs might belong to a new subclass of testis-specific proteins that act independently of the MAPK signal transduction cascade and do not depend on N-terminal docking regions for substrate binding.