Complex diffractive surface patterns on metals by UV-ps laser ablation.

Diffractive surface patterns with complex textures are generated on metal surfaces by picosecond UV laser ablation using an interference setup. Two diffraction gratings with variable distance and rotation angle provide a huge variety of interference patterns and thus resulting surface topographies. This variety can be further enhanced by selecting or blocking particular beams. A correlation analysis of the complex diffraction patterns generated by reflection of visible laser light at these surface topographies demonstrates that patterns with slightly differing fabrication parameters (variation of 0.5 mm in distance or 1° in rotation) can be clearly distinguished.

Hybrid micro-optical elements by laser-based fabrication of Fresnel lenses on the end face of gradient index lenses.

Fresnel lenses are fabricated directly upon the end face of gradient index (GRIN) lenses by F2-laser machining at 157 nm wavelength. The employed laser processing technique combines a mask projection configuration at 25-x demagnification with a rotation of the structured lens. The ablation characteristics of the GRIN materials require very high pulse fluences with typical values above 7 J/cm2. Topography measurements on the Fresnel lenses reveal a good contour accuracy with residual deviations from the design profile well below 100 nm. Such hybrid optical elements, combining GRIN lenses with diffractive lenses in one element, can serve as the basis for high-performance micro-optical imaging systems with diameters up to 2 mm. Examples of possible applications include imaging sensors like proximity sensors or color-corrected microscope objectives with high numerical aperture for endoscopy applications.

Insight on agglomerates of gold nanoparticles in glass based on surface plasmon resonance spectrum: study by multi-spheres T-matrix method.

The formation of a localized surface plasmon resonance (SPR) spectrum of randomly distributed gold nanoparticles in the surface layer of silicate float glass, generated and implanted by UV ArF-excimer laser irradiation of a thin gold layer sputter-coated on the glass surface, was studied by the T-matrix method, which enables particle agglomeration to be taken into account. The experimental technique used is promising for the production of submicron patterns of plasmonic nanoparticles (given by laser masks or gratings) without damage to the glass surface. Analysis of the applicability of the multi-spheres T-matrix (MSTM) method to the studied material was performed through calculations of SPR characteristics for differently arranged and structured gold nanoparticles (gold nanoparticles in solution, particles pairs, and core-shell silver-gold nanoparticles) for which either experimental data or results of the modeling by other methods are available. For the studied gold nanoparticles in glass, it was revealed that the theoretical description of their SPR spectrum requires consideration of the plasmon coupling between particles, which can be done effectively by MSTM calculations. The obtained statistical distributions over particle sizes and over interparticle distances demonstrated the saturation behavior with respect to the number of particles under consideration, which enabled us to determine the effective aggregate of particles, sufficient to form the SPR spectrum. The suggested technique for the fitting of an experimental SPR spectrum of gold nanoparticles in glass by varying the geometrical parameters of the particles aggregate in the recurring calculations of spectrum by MSTM method enabled us to determine statistical characteristics of the aggregate: the average distance between particles, average size, and size distribution of the particles. The fitting strategy of the SPR spectrum presented here can be applied to nanoparticles of any nature and in various substances, and, in principle, can be extended for particles with non-spherical shapes, like ellipsoids, rod-like and other T-matrix-solvable shapes.

Additivity of adrenaline and contractions on hormone-sensitive lipase, but not on glycogen phosphorylase, in rat muscle.

AIM: Hormone-sensitive lipase (HSL) has been proposed to regulate triacylglycerol (TG) breakdown in skeletal muscle. In muscles with different fibre type compositions the influence on HSL of two major stimuli causing TG mobilization was studied. METHODS: Incubated soleus and extensor digitorum longus (EDL) muscles from 70 g rats were stimulated by adrenaline (5.5 microm, 6 min) or contractions (200 ms tetani, 1 Hz, 1 min) in maximally effective doses or by both adrenaline and contractions. RESULTS: Hormone-sensitive lipase activity was increased significantly by adrenaline as well as contractions, and the highest activity (P < 0.05) was seen with combined stimulation [Soleus: 0.40 +/- 0.03 (SE) m-unit mg protein(-1) (basal), 0.65 +/- 0.02 (adrenaline), 0.65 +/- 0.03 (contractions), 0.78 +/- 0.03 (adrenaline and contractions); EDL: 0.18 +/- 0.01, 0.30 +/- 0.02, 0.26 +/- 0.02, 0.32 +/- 0.01]. Glycogen phosphorylase activity was always increased more by adrenaline compared with contractions [Soleus: 60 +/- 4 (a/a + b)% vs. 46 +/- 3 (P < 0.05); EDL: 60 +/- 5 vs. 39 +/- 6 (P < 0.05)]. After combined stimulation glycogen phosphorylase activity in soleus [59 +/- 3 (a/a + b)%] was identical to and in EDL [45 +/- 4 (a/a + b)%] smaller (P < 0.05) than the activity after adrenaline only. CONCLUSIONS: In slow-twitch oxidative as well as in fast-twitch glycolytic muscle HSL is activated by both adrenaline and contractions. These stimuli are partially additive indicating at least partly different mechanisms of action. Contractions may impair the enhancing effect of adrenaline on glycogen phosphorylase activity in muscle.

Effect of force development on contraction induced glucose transport in fast twitch rat muscle.

A previous study has shown that in fast twitch frog sartorius muscle contraction stimulated glucose transport depends only on stimulation frequency and not on workload. In contrast, we have recently shown that in rat slow twitch muscle stimulated to contract at constant frequency, glucose transport varies directly with force development and, in turn, metabolism. The present study was carried out to clarify whether the discrepancy between the earlier studies reflected differences in physiological behaviour between fast and slow twitch muscle. We investigated the effect of force development on glucose transport in incubated fast twitch rat flexor digitorum brevis (rich in type 2a fibres) and epitrochlearis (rich in type 2b fibres) muscle. Muscles were electrically stimulated to perform repeated tetanic contractions at 1 Hz for 10 min. Resting length was adjusted to achieve either no force or maximum force. Glucose transport (2-deoxyglucose uptake) increased when force was produced compared with when it was not (P < 0.05) in both flexor digitorum brevis (19 +/- 7 (basal), 163 +/- 14 (no force) and 242 +/- 17 (max force) nmol x g(-1) x 5 min(-1)) and epitrochlearis (60 +/- 4 (basal), 100 +/- 7 (no force) and 125 +/- 6 (max force) nmol x g(-1) x 5 min(-1)). In both muscles glucose transport increased in parallel with metabolic rate, as reflected by muscle lactate concentrations and 5' AMP-activated protein kinase activity, during contractions. In conclusion, as previously shown for rat soleus muscle, at a given stimulation frequency glucose transport varies directly with force development in rat flexor digitorum brevis and epitrochlearis muscle. Accordingly, force development enhances glucose transport in all mammalian muscle fibre types. The influence of force development probably reflects effects of enhanced 5' AMP-activated protein kinase activity resulting from reduced intra-cellular energy status and pH.

Adrenaline and glycogenolysis in skeletal muscle during exercise: a study in adrenalectomised humans.

The role of adrenaline in regulating muscle glycogenolysis and hormone-sensitive lipase (HSL) activity during exercise was examined in six adrenaline-deficient bilaterally adrenalectomised, adrenocortico-hormonal-substituted humans (Adr) and in six healthy control individuals (Con). Subjects cycled for 45 min at approximately 70% maximal pulmonary O2 uptake (VO2,max) followed by 15 min at approximately 86% VO2,max either without (-Adr and Con) or with (+Adr) adrenaline infusion that elevated plasma adrenaline levels (45 min, 4.49+/-0.69 nmol l(-1); 60 min, 12.41+/-1.80 nmol l(-1)). Muscle samples were obtained at 0, 45 and 60 min of exercise. In -Adr and Con, muscle glycogen was similar at rest (-Adr, 409+/-19 mmol (kg dry wt)(-1); Con, 453+/-24 mmol (kg dry wt)(-1)) and following exercise (-Adr, 237+/-52 mmol (kg dry wt)(-1); Con, 227+/-50 mmol (kg dry wt)(-1)). Muscle lactate, glucose-6-phosphate and glucose were similar in -Adr and Con, whereas glycogen phosphorylase (a/a + b x 100 %) and HSL (% phosphorylated) activities increased during exercise in Con only. Adrenaline infusion increased activities of phosphorylase and HSL as well as blood lactate concentrations compared with those in -Adr, but did not enhance glycogen breakdown (+Adr, glycogen following exercise: 274+/-55 mmol (kg dry wt)(-1)) in contracting muscle. The present findings demonstrate that during exercise muscle glycogenolysis can occur in the absence of adrenaline, and that adrenaline does not enhance muscle glycogenolysis in exercising adrenalectomised subjects. Although adrenaline increases the glycogen phosphorylase activity it is not essential for glycogen breakdown in contracting muscle. Finally, a novel finding is that the activity of HSL in human muscle is increased in exercising man and this is due, at least partly, to stimulation by adrenaline.

Effect of stimulation frequency on contraction-induced glucose transport in rat skeletal muscle.

Previous studies have indicated that frequency of stimulation is a major determinant of glucose transport in contracting muscle. We have now studied whether this is so also when total force development or metabolic rate is kept constant. Incubated soleus muscles were electrically stimulated to perform repeated tetanic contractions at four different frequencies (0.25, 0.5, 1, and 2 Hz) for 10 min. Resting length was adjusted to achieve identical total force development or metabolic rate (glycogen depletion and lactate accumulation). Overall, at constant total force development, glucose transport (2-deoxyglucose uptake) increased with stimulation frequency (P < 0.05; basal: 25 +/- 2, 0.25 Hz: 50 +/- 4, 0.5 Hz: 50 +/- 3, 1 Hz: 81 +/- 5, 2 Hz: 79 +/- 3 nmol. g(-1). 5 min(-1)). However, glucose transport was identical (P > 0.05) at the two lower (0.25 and 0.5 Hz) as well as at the two higher (1 and 2 Hz) frequencies. Glycogen decreased (P < 0.05; basal: 19 +/- 1, 0.25 Hz: 13 +/- 1, 0.5 Hz: 12 +/- 2, 1 Hz: 7 +/- 1, 2 Hz: 7 +/- 1 mmol/kg) and 5'-AMP-activated protein kinase (AMPK) activity increased (P < 0. 05; basal: 1.7 +/- 0.4, 0.25 Hz: 32.4 +/- 7.0, 0.5 Hz: 36.5 +/- 2.1, 1 Hz: 63.4 +/- 8.0, 2 Hz: 67.0 +/- 13.4 pmol. mg(-1). min(-1)) when glucose transport increased. Experiments with constant metabolic rate were carried out in soleus, flexor digitorum brevis, and epitrochlearis muscles. In all muscles, glucose transport was identical at 0.5 and 2 Hz (P > 0.05); also, AMPK activity did not increase with stimulation frequency. In conclusion, muscle glucose transport increases with stimulation frequency but only in the face of energy depletion and increase in AMPK activity. This indicates that contraction-induced glucose transport is elicited by metabolic demands rather than by events occurring early during the excitation-contraction coupling.

Stimulation of hormone-sensitive lipase activity by contractions in rat skeletal muscle.

Because the enzymic regulation of muscle triglyceride breakdown is poorly understood we studied whether neutral lipase in skeletal muscle is activated by contractions. Incubated soleus muscles from 70 g rats were electrically stimulated for 60 min. Neutral lipase activity against triacylglycerol increased after 1 and 5 min of contractions [0.36 +/- 0.02 (basal) versus 0.49 +/- 0.05 (1 min) and 0.54 +/- 0.05 (5 min) m-unit.mg of protein(-1), means +/- S.E.M., P < 0.05]. After 10 min the neutral lipase activity (0.40 +/- 0.05 m-unit.mg of protein(-1)) had decreased to basal values (P > 0.05). The contraction-mediated increase in lipase activity was increased by approximately 110% when muscle was stimulated in the presence of okadaic acid. Conversely, treatment of muscle homogenate with alkaline phosphatase completely reversed the contraction-mediated lipase activation. Lipase activity did not change during contractions when analysed in the presence of anti-hormone-sensitive-lipase (HSL) antibody [0.17 +/- 0.02 (basal) versus 0.21 +/- 0.02 (5 min) m-unit.mg of protein(-1), P > 0.05]. Furthermore, immunoprecipitation with affinity-purified anti-HSL antibody reduced muscle-HSL protein concentration by 81+/-4% and caused similar reductions in lipase activity against triacylglycerol and in the contraction-induced increase in this activity. Neither prior sympathectomy [0.33+/- 0.02 (basal) versus 0.53 +/- 0.06 (5 min) m-unit.mg of protein(-1), P < 0.05] nor propranolol impaired the lipase response to contractions. Glycogen phosphorylase activity in the absence of AMP increased after 1 min [27.3 +/- 3.1 versus 8.9 +/- 1.8% (activity without AMP/total activity with AMP), P < 0.05] and returned to basal levels after 5 min. In conclusion, skeletal-muscle-immunoreactive HSL is transiently stimulated by contractions and the mechanism probably involves phosphorylation. The time course of HSL activation is similar to that of glycogen phosphorylase. Apparently, the two enzymes are regulated in parallel by contraction-induced as well as hormonal mechanisms, allowing simultaneous recruitment of all major extra- and intra-muscular energy stores.

Dissociation of AMP-activated protein kinase activation and glucose transport in contracting slow-twitch muscle.

5'AMP-activated protein kinase (AMPK) has been suggested to be a key regulatory protein in exercise signaling of muscle glucose transport. To test this hypothesis, we investigated whether muscle glycogen levels affect AMPK activation and glucose transport stimulation similarly during contractions. Rats were preconditioned by a combination of swimming exercise and diet to obtain a glycogen-supercompensated group (high muscle glycogen content [HG]) with approximately 3-fold higher muscle glycogen levels than a glycogen-depleted group (low muscle glycogen content [LG]). In perfused fast-twitch muscles, contractions induced significant increases in AMPK activity and glucose transport and decreases in acetyl-CoA carboxylase (ACC) activity in both HG and LG groups. Contraction-induced glucose transport was nearly 2-fold (P < 0.05) and AMPK activation was 3-fold (P < 0.05) higher in the LG group compared with the HG group, whereas ACC deactivation was not different between groups. Thus, there was a significant positive correlation between AMPK activity and glucose transport in contracting fast-twitch muscles (r = 0.80, P < 0.01). However, in slow-twitch muscles with HG, glucose transport was increased 6-fold (P < 0.05) during contractions, whereas AMPK activity did not increase. In contracting slow-twitch muscles with LG, the increase in AMPK activity (315%) and the decrease in ACC activity (54 vs. 34% at 0.2 mmol/l citrate, LG vs. HG) was higher (P < 0.05) compared with HG muscles, whereas the increase in glucose transport was identical in HG and LG. In conclusion, in slow-twitch muscles, high glycogen levels inhibit contraction-induced AMPK activation without affecting glucose transport. This observation suggests that AMPK activation is not an essential signaling step in glucose transport stimulation in skeletal muscle.

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.

Effect of tension on contraction-induced glucose transport in rat skeletal muscle.

We questioned the general view that contraction-induced muscle glucose transport only depends on stimulation frequency and not on workload. Incubated soleus muscles were electrically stimulated at a given pattern for 5 min. Resting length was adjusted to achieve either no force (0% P), maximum force (100% P), or 50% of maximum force (50% P). Glucose transport (2-deoxy-D-glucose uptake) increased directly with force development (P < 0.05) [27 +/- 2 (basal), 45 +/- 2 (0% P), 68 +/- 3 (50% P), and 94 +/- 3 (100% P) nmol. g(-1). 5 min(-1)]. Glycogen decreased at 0% P but did not change further with force development (P > 0.05). Lactate, AMP, and IMP concentrations were higher (P < 0.05) and ATP concentrations lower (P < 0.05) when force was produced than when it was not. 5'-AMP-activated protein kinase (AMPK) activity increased directly with force [20 +/- 2 (basal), 60 +/- 11 (0% P), 91 +/- 12 (50% P), and 109 +/- 12 (100% P) pmol. mg(-1). min(-1)]. Passive stretch (approximately 86% P) doubled glucose transport without altering metabolism. In conclusion, contraction-induced muscle glucose transport varies directly with force development and is not solely determined by stimulation frequency. AMPK activity is probably an essential determinant of contraction-induced glucose transport. In contrast, glycogen concentrations per se do not play a major role. Finally, passive stretch per se increases glucose transport in muscle.

Expression of hormone-sensitive lipase and its regulation by adrenaline in skeletal muscle.

The enzymic regulation of triacylglycerol breakdown in skeletal muscle is poorly understood. Western blotting of muscle fibres isolated by collagenase treatment or after freeze-drying demonstrated the presence of immunoreactive hormone-sensitive lipase (HSL), with the concentrations in soleus and diaphragm being more than four times the concentrations in extensor digitorum longus and epitrochlearis muscles. Neutral lipase activity determined under conditions optimal for HSL varied directly with immunoreactivity. Expressed relative to triacylglycerol content, neutral lipase activity in soleus muscle was about 10 times that in epididymal adipose tissue. In incubated soleus muscle, both neutral lipase activity against triacylglycerol (but not against a diacylglycerol analogue) and glycogen phosphorylase activity increased in response to adrenaline (epinephrine). The lipase activation was completely inhibited by anti-HSL antibody and by propranolol. The effect of adrenaline could be mimicked by incubation of crude supernatant from control muscle with the catalytic subunit of cAMP-dependent protein kinase, while no effect of the kinase subunit was seen with supernatant from adrenaline-treated muscle. The results indicate that HSL is present in skeletal muscle and is stimulated by adrenaline via beta-adrenergic activation of cAMP-dependent protein kinase. The concentration of HSL is higher in oxidative than in glycolytic muscle, and the enzyme is activated in parallel with glycogen phosphorylase.

Hormone-sensitive lipase (HSL) expression and regulation in skeletal muscle.

Because the enzymatic regulation of muscle triglyceride metabolism is poorly understood we explored the character and activation of neutral lipase in muscle. Western blotting of isolated rat muscle fibers demonstrated expression of hormone-sensitive lipase (HSL). In incubated soleus muscle epinephrine increased neutral lipase activity by beta-adrenergic mechanisms involving cyclic AMP-dependent protein kinase (PKA). The increase was paralleled by an increase in glycogen phosphorylase activity and could be abolished by antiserum against HSL. Electrical stimulation caused a transient increase in activity of both neutral lipase and glycogen phosphorylase. The increase in lipase activity during contractions was not influenced by sympathectomy or propranolol. Training diminished the epinephrine induced lipase activation in muscle but enhanced the activation as well as the overall concentration of lipase in adipose tissue. In agreement with the in vitro findings, in adrenalectomized patients an increase in muscle neutral lipase activity was found at the end of prolonged exercise only if epinephrine was infused. In accordance with feedforward regulation of substrate mobilization in exercise, our studies have shown that HSL is present in skeletal muscle cells and is stimulated in parallel with glycogen phosphorylase by both epinephrine and contractions. HSL adapts differently to training in muscle compared with adipose tissue.

Optimization of a beam delivery system for a short-pulse KrF laser used for material ablation.

We present an optical arrangement for spatial homogenization of an UV beam carrying a short pulse (500 fs) to be used for material ablation. Conventional cylindrical fly's eye lens homogenizers (CFELH's) introduce unwanted interference effects into a beam caused by the high spatial coherence of short pulses. To prevent the disturbing effect of these intensity modulations, one can couple a low-loss distributed delay device to the CFELH. With the new design an intensity nonuniformity of <+/-5% rms can be obtained. High-resolution images of the beam profile show complete removal of the interference modulation. The pulse duration after homogenization is 12.5 ps. We performed preliminary ablation experiments in polyimide samples both by direct irradiation and by mask imaging. Uniformity and edge quality of the results are more than satisfactory, and the undesirable structure caused by interference is completely removed.