A prospective study of the association between smoking and later alcohol drinking in the general population

Aims To address the possible prospective association between smoking habits and risk of later heavy drinking in the adult population. Design Pooled population‐based long‐term cohort studies with repeated assessments of smoking and alcohol habits. Setting Copenhagen, Denmark. Participants A total of 14 130 non‐ to moderate drinkers at baseline, who attended re‐examination. Measurements Among the non‐ to moderate drinkers we addressed the relation between smoking habits at first examination and the risk of becoming a heavy and excessive drinker at follow‐up. Findings Level of tobacco consumption at first examination predicted an increased risk of becoming a heavy and excessive drinker in a dose‐dependent manner. Men who smoked more than 25 g of tobacco per day had adjusted odds ratios of 2.12 (95% confidence interval (CI): 1.44–3.11) and 3.95 (95% CI: 1.93–8.95) for becoming heavy and excessive drinkers, compared to participants who had never smoked. Equivalent estimates among women were 1.76 (95% CI: 1.02–3.04) and 2.21 (95% CI: 1.00–4.58), respectively. Conclusions This study suggests that tobacco use is associated quantitatively with later risk of heavier drinking.     

68Ga-labeling and in vivo evaluation of a uPAR binding DOTA- and NODAGA-conjugated peptide for PET imaging of invasive cancers

IntroductionThe urokinase-type plasminogen activator receptor (uPAR) is a well-established biomarker for tumor aggressiveness and metastatic potential. DOTA-AE105 and DOTA-AE105-NH₂ labeled with ⁶⁴Cu have previously been demonstrated to be able to noninvasively monitor uPAR expression using positron emission tomography (PET) in human cancer xenograft mice models. Here we introduce ⁶⁸Ga-DOTA-AE105-NH₂ and ⁶⁸Ga-NODAGA-AE105-NH₂ and evaluate their imaging properties using small-animal PET.MethodsSynthesis of DOTA-AE105-NH₂ and NODAGA-AE105-NH₂ was based on solid-phase peptide synthesis protocols using the Fmoc strategy. ⁶⁸GaCl₃ was eluted from a ⁶⁸Ge/⁶⁸Ga generator. The eluate was either concentrated on a cation-exchange column or fractionated and used directly for labeling. For in vitro characterization of both tracers, partition coefficient, buffer and plasma stability, uPAR binding affinity and cell uptake were determined. To characterize the in vivo properties, dynamic microPET imaging was carried out in nude mice bearing human glioma U87MG tumor xenograft.ResultsIn vitro experiments revealed uPAR binding affinities in the lower nM range for both conjugated peptides and identical to AE105. Labeling of DOTA-AE105-NH₂ and NODAGA-AE105-NH₂ with ⁶⁸Ga was done at 95°C and room temperature, respectively. The highest radiochemical yield and purity were obtained using fractionated elution, whereas a negative effect of acetone on labeling efficiency for NODAGA-AE105-NH₂ was observed. Good stability in phosphate-buffered saline and mouse plasma was observed. High cell uptake was found for both tracers in U87MG tumor cells. Dynamic microPET imaging demonstrated good tumor-to-background ratio for both tracers. Tumor uptake was 2.1% ID/g and 1.3% ID/g 30 min postinjection and 2.0% ID/g and 1.1% ID/g 60 min postinjection for ⁶⁸Ga-NODAGA-AE105-NH₂ and ⁶⁸Ga-DOTA-AE105-NH₂, respectively. A significantly higher tumor-to-muscle ratio (P<.05) was found for ⁶⁸Ga-NODAGA-AE105-NH₂ 60 min postinjection.ConclusionsThe use of ⁶⁸Ga-DOTA-AE105-NH₂ and ⁶⁸Ga-NODAGA-AE105-NH₂ as the first gallium-68 labeled uPAR radiotracers for noninvasive PET imaging is reported, which combine versatility with good imaging properties. These new tracers thus constitute an interesting alternative to the ⁶⁴Cu-labeled version (⁶⁴Cu-DOTA-AE105 and 64Cu-DOTA-AE105-NH₂) for detecting uPAR expression in tumor tissue. In our hands, the fractionated elution approach was superior for labeling of peptides, and ⁶⁸Ga-NODAGA-AE105-NH₂ is the favored tracer as it provides the highest tumor-to-background ratio.     

Calphostin C is an inhibitor of contraction, but not insulin-stimulated glucose transport, in skeletal muscle.

Wortmannin selectively impairs insulin-stimulated glucose transport in skeletal muscle. To search for an inhibitor specific for contraction-stimulated glucose transport, we screened a number of calmodulin and PKC inhibitors for their ability to impair contraction- and insulin-stimulated 2-deoxyglucose uptake in incubated rat soleus muscles. In concentrations that did not reduce contraction-induced force output, among calmodulin inhibitors W-7 inhibited both contraction- and insulin-stimulated glucose transport by up to 50% (P < 0.05), while Calmidazolium impaired only insulin-stimulated glucose transport (P < 0.05), and Trifluoperazine and Phenoxybenzamine did not influence glucose transport. In concentrations that did not reduce force generation, among PKC inhibitors Calphostin C specifically inhibited contraction-stimulated glucose transport (P < 0.05), whereas insulin-stimulated transport was impaired by Rottlerin and Bisindolylmaleimide I (P < 0.05), and both contraction- and insulin-stimulated glucose transport were inhibited by RO-31-8220 (P < 0.05). Calphostin C did not reduce contraction-induced increase in AMP-activated protein kinase (AMPK) activity. In conclusion, we have identified specific inhibitors of both contraction- and insulin-stimulated glucose transport. Both calmodulin and different isoenzymes of the PKC family may be involved in contraction- and insulin-stimulated glucose transport. Calphostin C does not influence glucose transport during contractions via stimulation of AMPK. Calphostin C may be used to unravel signal transduction in contraction-stimulated glucose transport.     

Male and female transsexualism: The danish experience with 37 patients

Since the first sex-reassignment operation in Denmark at the Rigshospitalet in 1951, a total of 37 patients, 29 males and 8 females, have had sexmodifying surgery and a change in legal status. In our experience a basic insecure gender identity is a predominant trait in transsexuals, dating back to earliest childhood. This insecurity and a concomitant anxiety are overcome differently by the two transsexual sexes. In male transsexualism, the most outstanding characteristic is a narcissistic withdrawal to a condition marked by submission and pseudofemininity. Anxiety and insecurity are basic to the gender dysphoria but are subdued by means of fantasy escape and gratification in aestheticized ego-ideals with suppression of aggressive and sexual feelings. This results in the often observed pseudofemininity in the male transsexual. A core group of transsexual males are marked by a persistent pseudofeminine narcissism. They have stable ego strength, are agenital in sexual attitude, and have an intact sense of reality. This group is expected to remain so after sex reassignment. The transsexual female assumes a narcissistic, phallic attitude displaying outer activities and caricatured masculine manners in an attempt to subdue her insecurity. Examples are given of the characteristic splitting of these persons' phenomenological egoexperiences and how different their reality testing is from that of psychotic persons with a desire for sex change. Transsexual females are much more sexually active than transsexual males. We find a closer connection between female homosexuality and transsexualism than between male homosexuality and transsexualism.     

Mechanisms associated with hypoxia- and contraction-mediated glucose transport in muscle are fibre-dependent

The purpose of this study was to examine the effects of hypoxia and muscle contractions on rates of 2-deoxyglucose (2-DG) transport in red and white portions of the gastrocnemius muscle of the rat. 2-DG transport was measured during the last 10 min of a 60-min hindlimb perfusion in male Wistar rats (≈ 300 g), with or without muscle contractions of one limb. The medium was gassed with either 95% oxygen and 5% carbon dioxide or 95% nitrogen and 5% carbon dioxide to achieve normal or hypoxic conditions, respectively. Muscle contractions began after 30 min of perfusion and consisted of isometric muscle actions (200-ms trains, 100 Hz; one train per second) for two sets of 5 min, with 1-min rest between sets. 2-DG transport in white gastrocnemius was higher (P < 0.05) than basal during hypoxia (4.8-fold) and following contractions using oxygenated or hypoxic medium (4.6-fold and 5.4-fold, respectively; n=6 for each group). 2-DG transport was not different (P > 0.05) between these stimulated conditions. Similarly, 2-DG transport in red gastrocnemius was 5.1- and 4.8-fold higher (P < 0.05) than basal during hypoxia and following contractions in oxygenated medium, respectively. However, 2-DG transport following contractions during hypoxic conditions in red gastrocnemius was, unlike white gastrocnemius, higher (8.9-fold over basal; P < 0.05) than in all other conditions. These results suggest that mechanisms associated with hypoxia- and muscle contraction-mediated glucose transport are fibre type-dependent, with additive effects of the two stimuli in fast-twitch, oxidative fibres.     

Imaging of insulin signaling in skeletal muscle of living mice shows major role of T-tubules

Insulin stimulates glucose transport in skeletal muscle by glucose transporter GLUT4 translocation to sarcolemma and membrane invaginations, the t-tubules. Although muscle glucose uptake plays a key role in insulin resistance and type 2 diabetes, the dynamics of GLUT4 translocation and the signaling involved are not well described. We have now developed a confocal imaging technique to follow trafficking of green fluorescent protein-labeled proteins in living muscle fibers in situ in anesthetized mice. Using this technique, by imaging the dynamics of GLUT4 translocation and phosphatidylinositol 3,4,5 P(3) (PIP(3)) production in response to insulin, here, for the first time, we delineate the temporal and spatial distribution of these processes in a living animal. We find a 10-min delay of maximal GLUT4 recruitment and translocation to t-tubules compared with sarcolemma. Time-lapse imaging of a fluorescent dye after intravenous injection shows that this delay is similar to the time needed for insulin diffusion into the t-tubule system. Correspondingly, immunostaining of muscle fibers shows that insulin receptors are present throughout the t-tubule system. Finally, PIP(3) production, an early event in insulin signaling, progresses slowly along the t-tubules with a 10-min delay between maximal PIP(3) production at sarcolemma compared with deep t-tubules following the appearance of dye-labeled insulin. Our findings in living mice indicate a major role of the t-tubules in insulin signaling in skeletal muscle and show a diffusion-associated delay in insulin action between sarcolemma and inner t-tubules.