Evidence that leptin contributes to intestinal cholesterol absorption in obese (ob/ob) mice and wild-type mice

In the present study, the effect of leptin on intestinal cholesterol absorption was investigated in C57 BL/6 OlaHsd Lep^ob/Lep^ob obese (ob/ob) mice and lean C57 BL/6 (wild-type) mice. Animals were treated either with or without recombinant leptin for 2 wk. Cholesterol absorption was measured by the constant isotope feeding method and indirectly by the ratio of campesterol to cholesterol in serum. In ob/ob mice, cholesterol absorption was significantly higher compared to wild-type mice [83.4±2.3% (SD) vs. 77.6±1.5%, P<0.01]. Treatment with leptin significantly reduced cholesterol absorption in both ob/ob and wild-type mice by 8.5 (P<0.001) and 5.2% (P<0.05), respectively. Serum concentrations of campesterol and the ratio of campesterol to cholesterol in ob/ob mice were significantly higher compared to wild-type mice (2.2±0.3 mg/dL vs. 1.2±0.3 mg/dL, P<0.001; and 36.8±2.8 μg/mg vs. 28.0±3.3 μg/mg, P<0.001). After treatment of ob/ob mice with leptin, concentrations of campesterol and its ratio to cholesterol were significantly lower (2.2±0.3 mg/dL vs. 1.0±0.2 μg/mg, P<0.001; and 36.8±2.8 μg/mg vs. 13.2±2.2 μg/mg, P<0.001, respectively). In wild-type mice, the ratio of campesterol to cholesterol in serum was also significantly lower after treatment with leptin (28.0±3.3 μg/mg vs. 22.6±5.0 μg/mg, P<0.05). A significant positive correlation (r=0.701, P<0.01) between cholesterol absorption and the ratio of campesterol to cholesterol, in serum was found. It is concluded that leptin contributes to intestinal cholesterol absorption in ob/ob mice and lean wild-type mice.     

Colony-blot assay with anti-p60 antibodies as a method for quick identification of Listeria in food

The present study evaluated the ability to isolate Listeria from foods, using shortened procedure of sample enrichment followed by immunomagnetic separation or filtration methods, and serological identification of isolated bacteria by colony-blot and Western blot methods with anti-p60 antibodies. By these rapid methods, identification of Listeria was achieved in much shorter time (40-48 h) than with standard cultivation and biochemical identification procedures. The rapid methods used are easy to perform and, what is most important, their specificity is very high and fulfills the expectations. The possibility to select Listeria colonies growing on non-selective media by blotting with anti-p60 antiserum seems to be particularly valuable in examination of food samples containing/not too many Listeria (1-10 CFU/25 g). However, the blot method using anti-PepD mAb specific to unique region of L. monocytogenes p60 is necessary to distinguish L. monocytogenes from other Listeria species.     

Wavelet decomposition of cardiovascular signals for baroreceptor function tests in pigs

In this paper, the discrete wavelet transform (DWT) was applied to analyze the fluctuations in RR interval and systolic arterial pressure (SAP) recorded from eight alpha-chloralose anesthetized pigs. Our aim was to characterize the autonomic modulation before and after cardiac autonomic blockade and during baroreflex function tests. The instantaneous power of decomposed low-frequency (LF) and high-frequency (HF) components was used for a time-variant spectral analysis. Our results suggested that transient events and changes in autonomic modulation were detected with high temporal resolution. A nonlinear relationship between RR interval and SAP during pharmacologically induced changes in blood pressure was found, when the superimposed effect of respiratory sinus arrhythmia was removed. In addition, the baroslopes were nearly linear when both the LF and HF components were removed using DWT decomposition.     

Closed-loop phase behaviour in block copolymers

Closed-loop phase diagrams are known in systems with specific intermolecular interactions. In weakly interacting systems, however, such behaviour has never been observed. Here, diblock copolymers formed from polystyrene covalently linked to poly (n-pentylmethacrylate), P(S-b-nPMA), which have only weak segmental interactions, are shown to exhibit a closed-loop phase behaviour over a narrow range of molecular weight. The endothermic transitions from the disordered to ordered and back to the disordered state, as a function of increasing temperature, are dominantly entropic in origin. The morphology and rheological properties of P(S-b-nPMA) undergo characteristic changes at the transitions. Whereas the disorder-to-order transition temperature increases with decreasing molecular weight, the order-to-disorder transition temperature decreases. At a limiting molecular weight, the closed-loop vanishes and no ordering occurs. These findings provide quantitative insight into an elusive transition in weakly interacting multicomponent systems.     

Biosynthesis of novel thermoplastic polythioesters by engineered Escherichia coli

The development of non-petrochemical sources for the plastics industry continues to progress as large multinationals focus on renewable resources to replace fossil carbon. Many bacteria are known to accumulate polyoxoesters as water-insoluble granules in the cytoplasm. The thermoplastic and/or elastomeric behaviour of these biodegradable polymers holds promise for the development of various technological applications. Here, we report the synthesis and characterization of microbial polythioesters (PTEs), a novel class of biopolymers of general technological relevance. Biosynthesis of PTE homopolymers was achieved using a recombinant strain of Escherichia coli that expressed a non-natural pathway consisting of a butyrate kinase, a phosphotransbutyrylase, and a PHA synthase. Different homopolymers were produced, consisting of either 3-mercaptopropionate, 3-mercaptobutyrate, or 3-mercaptovalerate repeating units, if the respective mercaptoalkanoic acids were provided as precursor substrates to the fermentative process. The PTEs contributed up to 30% (w/w) of the cellular dry weight and were identified as hydrophobic inclusions in the cytoplasm. The chemical and stereochemical homogeneity of the purified PTEs were identified by different methods, and the estimated physical properties were compared to the oxypolyester equivalents, revealing low crystalline order and, for the poly(3-mercaptopropionate) improved thermal stability. The ability to produce PTEs through a biosynthetic route opens up new avenues in the field of biomaterials.     

Capacitive micromachined ultrasonic transducers: next-generation arrays for acoustic imaging?

Piezoelectric materials have dominated the ultrasonic transducer technology. Recently, capacitive micromachined ultrasonic transducers (CMUTs) have emerged as an alternative technology offering advantages such as wide bandwidth, ease of fabricating large arrays, and potential for integration with electronics. The aim of this paper is to demonstrate the viability of CMUTs for ultrasound imaging. We present the first pulse-echo phased array B-scan sector images using a 128-element, one-dimensional (1-D) linear CMUT array. We fabricated 64- and 128-element 1-D CMUT arrays with 100% yield and uniform element response across the arrays. These arrays have been operated in immersion with no failure or degradation in performance over the time. For imaging experiments, we built a resolution test phantom roughly mimicking the attenuation properties of soft tissue. We used a PC-based experimental system, including custom-designed electronic circuits to acquire the complete set of 128 x 128 RF A-scans from all transmit-receive element combinations. We obtained the pulse-echo frequency response by analyzing the echo signals from wire targets. These echo signals presented an 80% fractional bandwidth around 3 MHz, including the effect of attenuation in the propagating medium. We reconstructed the B-scan images with a sector angle of 90 degrees and an image depth of 210 mm through offline processing by using RF beamforming and synthetic phased array approaches. The measured 6-dB lateral and axial resolutions at 135 mm depth were 0.0144 radians and 0.3 mm, respectively. The electronic noise floor of the image was more than 50 dB below the maximum mainlobe magnitude. We also performed preliminary investigations on the effects of crosstalk among array elements on the image quality. In the near field, some artifacts were observable extending out from the array to a depth of 2 cm. A tail also was observed in the point spread function (PSF) in the axial direction, indicating the existence of crosstalk. The relative amplitude of this tail with respect to the mainlobe was less than -20 dB.     

Histomorphological, histomorphometrical and biomechanical analysis of ceramic bone substitutes in a weight-bearing animal model

It was the purpose of this investigation to prove the biomechanical properties, the osteoconductive capacity and the degradation rate of tricalcium phosphate ( TCP), a neutralized glass ceramics (GB9N) and a composite material (GB9N+copolymers). In a weight-bearing animal model six substitutes each were implanted in the medial tibial head of the right lower leg of adult Merino-sheep in a standardized surgical technique. After nine months the implants were harvested and prepared for histomorphological and histomorphometrical investigations (undecalcified Masson Goldner staining). For additional biomechanical testing of the specimens, non-operated bone blocks from the contralateral tibia as well as native implants served as controls. No significant differences for the maximum fracture load as well as for the yield strength were detected between harvested specimens and bone blocks from the contralateral tibia. However there were marked differences to ceramics that were not implanted. All substitutes showed osteoconduction, leading to a continuous ingrowth of new formed bone. However in the composite material soft tissue could be identified within the scaffold and there were signs of ongoing bone remodeling, nine months after implantation. The bone per tissue volume of -TCP in conjunction to new bone (=percentage of trabecular bone volume plus percentage of residual substitute) was higher than for GB9N and the composite material. Nine months after implantation the percentage of residual -TCP was 48%, it was 32% for GB9N and 28% for the composite.The intention of further studies should be to accelerate the degradation rates of substitutes and to improve biomechanical properties of implants by either modifying the chemical composition or combining materials with agents as, e.g. growth factors.     

Characterization of silica-based monoliths with bimodal pore size distribution

Band dispersion was studied and the retention thermodynamics addressed for insulin and angiotensin II on C18 silica monoliths with a bimodal pore size distribution, covering linear mobile-phase velocities up to 1 cm/s and different temperatures. These data suggest that the influence of average column pressure on retention (between 0 and 10 MPa) is not negligible. Plate height curves were interpreted with the van Deemter equation by assuming an independent contribution from mechanical and non-mechanical dispersion mechanisms. This analysis revealed diffusion-limited mass transfer in the mesoporous silica skeleton which, in turn, allowed us to calculate an equivalent dispersion particle diameter (d(disp) = 3 microm) using the C-term parameter of the van Deemter equation. The resulting superposition of reduced plate height curves for monolithic and particulate beds confirmed that this view presents an adequate analogy. The macroporous interskeleton network responsible for the hydraulic permeability of a monolith was translated to the interparticle pore space of particulate beds, and an equivalent permeability particle diameter (d(perm) = 15 microm) was obtained by scaling based on the Kozeny-Carman equation.     

Off-axis measurements of atmospheric trace gases by use of an airborne ultraviolet-visible spectrometer

An airborne UV-visible spectrometer, the Gas Analyzer Spectrometer Correlating Optical Differences, airborne version (GASCOD/A4pi) was successfully operated during the Airborne Polar Experiment, Geophysica Aircraft in Antarctica airborne campaign from Ushuaia (54 degrees 49' S, 68 degrees 18' W), Argentina in southern spring 1999. The instrument measured scattered solar radiation through three optical windows with a narrow field of view (FOV), one from the zenith, two from the horizontal, as well as actinic fluxes through 2pi FOV radiometric heads. Only a few airborne measurements of scattered solar radiation at different angles from the zenith are available in the literature. With our configuration we attempted to obtain the average line-of-sight concentrations of detectable trace gases. The retrieval method, based on differential optical absorption spectroscopy, is described and results for ozone are shown and compared with measurements from an in situ instrument as the first method of validation.     

All-solid-state tunable continuous-wave ultraviolet source with high spectral purity and frequency stability

We present a novel approach for the generation of higly frequency-stable, widely tunable, single-frequency cw UV light that is suitable for high-resolution spectroscopy. Sum-frequency generation (SFG) of two solid-state sources with a single cavity resonant for both fundamental waves is employed. Using a highly stable, narrow-linewidth frequency-doubled cw Nd:YAG laser as a master laser and slaving to it the SFG cavity and the other fundamental wave from a Ti:sapphire laser, we generate UV radiation of 33-mW output power around 313 nm. Alternatively, we use a diode laser instead of the Ti:sapphire laser and produce an output power of 2.1 mW at 313 nm. With both setups we obtain a continuous tunability of >15 GHz, short-term frequency fluctuations in the submegahertz range, a long-term frequency drift below 100 MHz/h, and stable operation for several hours. The theory of optimized doubly resonant SFG is also given.