Contractile tension and beating rates of self-exciting monolayers and 3D-tissue constructs of neonatal rat cardiomyocytes.

The CellDrum technology (The term 'CellDrum technology' includes a couple of slightly different technological setups for measuring lateral mechanical tension in various types of cell monolayers or 3D-tissue constructs) was designed to quantify the contraction rate and mechanical tension of self-exciting cardiac myocytes. Cells were grown either within flexible, circular collagen gels or as monolayer on top of respective 1-mum thin silicone membranes. Membrane and cells were bulged outwards by air pressure. This biaxial strain distribution is rather similar the beating, blood-filled heart. The setup allowed presetting the mechanical residual stress level externally by adjusting the centre deflection, thus, mimicking hypertension in vitro. Tension was measured as oscillating differential pressure change between chamber and environment. A 0.5-mm thick collagen-cardiac myocyte tissue construct induced after 2 days of culturing (initial cell density 2 x 10(4) cells/ml), a mechanical tension of 1.62 +/- 0.17 microN/mm(2). Mechanical load is an important growth regulator in the developing heart, and the orientation and alignment of cardiomyocytes is stress sensitive. Therefore, it was necessary to develop the CellDrum technology with its biaxial stress-strain distribution and defined mechanical boundary conditions. Cells were exposed to strain in two directions, radially and circumferentially, which is similar to biaxial loading in real heart tissues. Thus, from a biomechanical point of view, the system is preferable to previous setups based on uniaxial stretching.

Laser spectroscopy of niobium fission fragments: first use of optical pumping in an ion beam cooler buncher.

A new method of optical pumping in an ion beam cooler buncher has been developed to selectively enhance ionic metastable state populations. The technique permits the study of elements previously inaccessible to laser spectroscopy and has been applied here to the study of Nb. Model independent mean-square charge radii and nuclear moments have been studied for ;{90,90 m,91,91 m,92,93,99,101,103}Nb to cover the region of the N=50 shell closure and N approximately 60 sudden onset of deformation. The increase in mean-square charge radius is observed to be less than that for Y, with a substantial degree of beta softness observed before and after N=60.

The fixation strength of hydroxyapatite-coated Schanz screws and standard stainless steel Schanz screws in lower extremity lengthening : a comparison based on a new torque value index: the fixation index.

INTRODUCTION: Lengthening procedures are often complicated by loosening of pins. It has been reported that coating with hydroxyapatite improves fixation and reduces the rate of pin-track infection. MATERIALS AND METHODS: We compared 47 hydroxyapatite-coated Schanz screws (HA screws) in 12 monolateral fixators mounted at the University Hospital Hamburg-Eppendorf with 45 standard stainless steel screws in 9 monolateral fixators mounted at the St Josefs-Hospital Wiesbaden by measuring the insertion and extraction torque values. The average implantation period was 7 months for the hydroxyapatite-coated screws and 5.4 months for the uncoated screws. We established the quotient of the maximum extraction torque over insertion torque which shows the change in the fixation strength with respect to time, the fixation index. It eliminates the influence of the varying pin-bone contact. RESULTS: There was no significant difference in the rate of infection. In the Schanz screws without signs of infection the index was 1.92 for the HA screws and 0.76 for the stainless steel screws (P = 0.0002) giving evidence of the improvement of the fixation by the coating. CONCLUSION: HA coating resulted in improved fixation of Schanz screws in bone and may be useful in prolonged external fixation of the lower leg. The fixation index proved to be a simple tool for the evaluation of the fixation strength of Schanz screws.

Macular circulation in patients with diabetes mellitus with and without arterial hypertension.

BACKGROUND: Previous fluorescein angiographic studies have shown alterations in the macular microcirculation in patients with diabetes mellitus and arterial hypertension. In both diseases capillary blood velocity was reduced and capillary density decreased. These changes were more pronounced in diabetic patients. We have examined the influence of arterial hypertension in combination with diabetes mellitus. METHODS: 62 patients with diabetes mellitus and arterial hypertension (group 1) were matched with patients with diabetes mellitus but without arterial hypertension (group 2, match criteria: ETDRS stage of retinopathy). In all subjects fluorescein angiograms were performed with a scanning laser ophthalmoscope. Macular capillary blood velocity (CBV), perifoveal intercapillary area (PIA), the coefficient of variation of both parameters, the area of the foveal avascular zone (FAZ), and the arteriovenous passage time (AVP) were assessed by digital image analysis. RESULTS: Systolic and diastolic blood pressures were significantly increased in the patients with arterial hypertension (systolic p=0.0008; diastolic p=0.03). Neither dynamic measures (AVP: 1.64 (0.49) seconds (group 1), 1.72 (0.58) seconds (group 2); CBV: 1.98 (0.39) mm/s (group 1), 2.09 (0.43) mm/s (group 2)) nor morphological measures (PIA: 7985 (3137) microm(2) (group 1), 8338 (3376) microm(2) (group 2); FAZ: 0.319 (0.206) mm(2) (group 1), 0.363 (0.237) mm(2) (group 2)) were significantly different between the two groups of diabetic patients. CONCLUSION: Arterial hypertension did not result in more severe macular capillary dropout than diabetes without hypertension. This might be explained by the fact that most of the patients were being treated with antihypertensive drugs.

Immunfluorescence study of neuropeptides in identified neurons of the rat auditory superior olivary complex.

The present study was conducted to investigate the distribution and immunohistochemical characteristics of ascending and descending projection neurons of the rat superior olivary complex (SOC), a group of interrelated brainstem nuclei. Ascending neurons were identified by injection of cholera toxin B subunit (CTB) into the central nucleus of the inferior colliculus (IC), descending neurons were labeled by application of Fluoro-Gold (FG) into the scala tympani of the cochlea, ipsilaterally to the IC injection. In accordance with the literature, we observed neurons innervating the IC located in the lateral superior olivary nucleus (LSO) and dorsal periolivary groups (DPO) on both sides, in the superior paraolivary nucleus (SPO) predominantly ipsilateral, as well as in the ipsilateral medial superior olivary nucleus (MSO) and the medial nucleus of the trapezoid body (MNTB). Cochlear projection neurons were found predominantly in the ipsilateral LSO as well as in the bilateral SPO, DPO, MSO and MNTB. In addition, a considerable population of neurons in the ipsilateral LSO and SPO were identified as being both ascending and descending. To further characterize these double-projecting neurons, brainstem sections were incubated in antisera directed against different neuroactive substances. The majority of ascending/descending cells in the LSO contained calcitonin gene-related peptide, but not substance P (SP), met-enkephalin (ENK) or tyrosine hydroxylase (TH). Some of these neurons apparently were contacted by ENK- or SP-immunoreactive fibers and terminals. In addition, we found TH-immunoreactive neurons in the lateral MNTB region. These neurons, which were labeled upon tracer injection into the cochlea (but not upon IC injection), probably belong to the C1 catecholaminergic cell group and may represent a division of the uncrossed olivocochlear bundle. The present results reveal the existence of a previously unknown subpopulation of SOC neurons that project to both the cochlea and the inferior colliculus. Their CGRP immunoreactivity and their uncrossed projection pattern provide evidence that they may belong to the cholinergic, putatively excitatory cell group.

Evidence against specific binding of salicylic acid to plant catalase.

It was demonstrated that salicylic acid (SA) not only binds to catalase from differentiated higher plants and plant cell suspension cultures but also to those of fungi and animals. SA bound specifically to iron-containing enzymes, such as catalase, aconitase, lipoxidase and peroxidase, while not to iron-free plant enzymes. On the grounds of these experiments, the claim is further challenged that SA is a signalling compound and second messenger in plants that activates plant defense-related genes through elevated H2O2 levels by specifically inhibiting catalase activity. SA may just function as a phytoalexin.

Partial Purification and Properties of (S)-Norlaudanosoline Synthase from Eschscholtzia tenuifolia Cell Cultures.

A new enzyme, (S)-norlaudanosoline synthase, which catalyses the synthesis of (S)-norlaudanosoline from dopamine and 3,4-dihydroxyphenylacetaldehyde was isolated from the soluble protein extract of ESCHSCHOLTZIA TENUIFOLIA cell suspension cultures and purified approximately 40-fold. The apparent molecular weight of the enzyme is 15 500 Dalton. The pH optimum is 7.8, temperature optimum 40 degrees C, apparent K (M) values for dopamine and dihydroxyphenyl-acetaldehyde are 1.5 mM and 0.7 mM respectively. The synthase shows high substrate specificity in that only the phenylacetaldehydes are transformed but not the phenylpyruvates. No apparent cofactor requirement could be demonstrated. By means of isoelectric focusing and disc-gel electrophoresis evidence was obtained for the existence of four norlaudanosoline synthase isoenzymes, none of which catalyses the reaction of dopamine with 3,4-dihydroxyphenylpyruvate. These enzymes are responsible for the synthesis of (S)-norlaudanosoline, the key intermediate in the formation of isoquinoline alkaloids occurring in the plant kingdom.