Nanostructuring discotic molecules on ITO support.

Patterning of organic compounds on a nanometer length scale is of great interest for solar applications: defined control over the donor-acceptor interface will allow design of an optimized nano-morphology promoting exciton separation and reducing charge recombination. Herein we present an imprinting technique using anodized alumina oxide (AAO) hard templates as stamps. We show an exact pattern transfer of the AAO structures into a solution processable hexa-peri-hexabenzocoronene (HBC), a discotic small molecule with acrylate moieties which is polymerized in situ. Film fabrication is realized for a variety of nanowire dimensions on square centimeter areas. The fabrication directly on conductive glass support and control over the formation of a dense barrier layer render this approach appealing for the fabrication of fully organic nanostructured photovoltaic devices.

Regulation of aspartate-derived amino acid homeostasis in potato plants (Solanum tuberosum L.) by expression of E. coli homoserine kinase.

The availability of the carbon backbone O-phosphohomoserine (OPHS) is critical to methionine (met) and threonine (thr) synthesis. OPHS derives from homoserine and is formed by homoserine kinase (HSK). To clarify the function of HSK in cellular metabolism, the E. coli HSK ortholog thrB was expressed in potato plants targeting the EcHSK protein to chloroplasts and to the cytosol. Both approaches resulted in up to 11 times increased total HSK enzyme activity. Transgenic plants exhibited reduced homoserine levels while met and thr did not accumulate significantly. However, the precursor cysteine and upstream intermediates of met such as cystathionine and homocysteine did indicating an accelerated carbon flow towards the end products. Coincidently, plants with elevated cytosolic levels of EcHSK exhibited a reduction in transcript levels of the endogenous HSK, as well as of threonine synthase (TS), cystathionine beta-lyase (CbL), and met synthase (MS). In all plants, cystathionine gamma-synthase (CgS) expression remained relatively unchanged from wild type levels, while S-adenosylmethionine synthetase (SAMS) expression increased. Feeding studies with externally supplied homoserine fostered the synthesis of met and thr but the regulation of synthesis of both amino acids retained the wild type regulation pattern. The results indicate that excess of plastidial localised HSK activity does not influence the de novo synthesis of met and thr. However, expression of HSK in the cytosol resulted in the down-regulation of gene expression of pathway genes probably mediated via OPHS. We integrated these data in a novel working model describing the regulatory mechanism of met and thr homeostasis.

Glucocorticoid receptor binding site in the mouse alpha-amylase 2 gene mediates response to the hormone.

Amylase gene expression has been shown to be positively regulated by glucocorticoids. Previous reports have suggested that this effect is indirect. We have addressed this question in a mouse exocrine pancreas cell line, 266-6, in which basal level of expression of amylase mRNA is low but inducible by glucocorticoids. In these cells the effect of glucocorticoids is not inhibited by cycloheximide at early time points. Reporter plasmids containing 224 base pairs of mouse amylase 5'-flanking DNA are positively regulated by glucocorticoids in gene transfer experiments. Glucocorticoid receptor purified from rat liver binds to the amylase promoter from position -56 to -33 and at the start of transcription. Site-directed mutation at the upstream position (-47 to -42) eliminates response to glucocorticoids in transient gene transfer experiments. Thus, glucocorticoid regulation of the mouse amylase gene is a direct effect and is mediated via a receptor binding site in the promoter region of the gene. Inhibition of the hormone response by cycloheximide at later time points after induction suggests the additional requirement for a short-lived factor. The DNA binding domain of the glucocorticoid receptor binds to a single site in the amylase promoter as a monomer, suggesting that both receptor binding sites as well as an additional short-lived factor are required to obtain induction.

Central somatosensory conduction time in severely growth-stunted children.

To examine the effects of chronic malnutrition on central nervous system function, we used the somatosensory evoked potential to measure the central conduction time of 20 children aged 7-8 y with heights below the third percentile for their age and 20 control children in Honduras. The two groups differed significantly in socioeconomic status, achievement in Bender's neurointegrative test, and hematocrit, but not in birth weight. After median nerve stimulation, the mean central conduction time (interpeak latency between N13 and N20) for the growth-stunted group (6.19 +/- 0.52 ms) did not differ significantly from that of the control subjects (6.30 +/- 0.58 ms), suggesting appropriate myelination and fiber diameter. Somatosensory tracts may escape damage resulting from postnatal dietary deficiencies because myelination in these tracts is almost complete at birth.

Ciliary beat frequency of biopsy samples taken from normal persons and patients with various lung diseases.

Ciliary beat frequency from human tracheobronchial mucosa was measured in vitro. Biopsy samples were taken during bronchoscopy and beat frequency measured with a phase contrast microscope and a photosensitive cell. In 20 healthy volunteers the mean ciliary beat frequency in the trachea was 12.5 +/- 2.8 Hz at 37 degrees C with a wide interindividual range (4.9 to 17.4 Hz), but a good intraindividual reproducibility (+/- 7 percent). There was a slight increase in frequency down the tracheobronchial tree and a decrease at lower temperatures. With some exceptions, no disease-specific correlation in beat frequency seems to exist in different bacterial and malignant bronchopulmonary conditions. However the quantity and quality of ciliated cells brushed from the mucosa does appear to be disease-related.

Indirect RT-PCR in-situ hybridization: a novel non-radioactive method for detecting glucose-dependent insulinotropic peptide.

To establish indirect in-situ PCR for the detection of intestinal peptide hormones, rat intestine and a murine intestinal tumor cell line, STC 1, were used. The results exhibited intensive staining of GIP-producing K-cells. Paraformaldehyde-fixed cryostat sections yielded the best results in signal to background ratio with RT-PCR in-situ hybridization. Moreover, it was possible to elevate the positive staining signal and to reduce background staining. Digoxigenin-labeled in-situ hybridization served as a control for specificity and sensitivity of GIP (glucose-dependent insulinotropic peptide) mRNA expression on cryostat as well as paraffin sections. In conclusion, this RT-PCR in-situ hybridization protocol proves to be a specific, sensitive and reliable non-radioactive technique for the detection of intestinal peptide hormone mRNA, especially in tissues or tumor cells where the application of ISH is limited.

Early malnutrition followed by nutritional restoration lowers the conduction velocity and excitability of the corticospinal tract.

The physiological sequelae of undernutrition were investigated in rats that were undernourished from day 1-21 and subsequently free-fed to 75 days of age. Population responses were recorded in the corticospinal tract following surface stimulation of the motor cortex, which activates corticospinal cells directly, and also indirectly via cortical synapses. The conduction velocity of the fastest corticospinal fibers in 15 malnourished rats was 16.9 m/s, significantly slower (P < 0.001) than the 20.0 m/s observed in 26 controls. In addition, the excitability of corticospinal neurons to direct stimulation was reduced as much as 67% in malnourished rats, while no effect on synaptic activation was observed. Our findings suggest that early malnutrition reduces the number of large fibers in the adult corticospinal tract. These results are discussed with respect to known morphological and behavioral effects of malnutrition in rats and their relevance to humans.

"Human pancreas-specific protein" (procarboxypeptidase B): a valuable marker in pancreatitis?

Human pancreas-specific protein (PASP) has been characterized previously as a serum marker for pancreatitis. It was then identified as pancreatic procarboxypeptidase B (PCB). The aim of the present study was to verify the usefulness of PASP (PCB) as a serum marker in patients with acute (n = 20) and chronic (n = 12) pancreatitis and in those following endoscopic retrograde cholangiopancreaticography (ERCP) (n = 44). Serum PASP values were analyzed by radioimmunoassay, with a range of normal values between 15 and 111 ng/ml. Between April 1992 and September 1992, 20 subjects (19-86 years of age) with acute pancreatitis (alcoholic, 8; biliary, 8; other, 4) were studied. We found edematous pancreatitis in 17 cases and severe hemorrhagic pancreatitis in three cases. At admission, peak levels of PASP (average value, 1,976 +/- 329 ng/ml), pancreatic isoamylase (942 +/- 151 U/L) and lipase (2,946 +/- 534 U/L) were detected in 15 of 20, 16 of 20, and 12 of 20 cases, respectively. The etiology of the pancreatitis had no influence on the PASP values. Furthermore, 10 patients with alcoholic and two patients with nonalcoholic chronic pancreatitis (29-67 years of age) were studied. The average peak level of PASP was 1,229 +/- 434 ng/ml. In this group, PASP paralleled the time course of amylase and lipase. Maximal PASP, amylase, and lipase levels were found in 11 of 12, nine of 12, and five of 12 patients, respectively, on the day of admission. ERCP was performed in 44 patients (36-87 years of age), demonstrating common bile duct stones in 16 and bile or pancreatic ductal tumors in 15 cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Brachial and digital artery pulse pressures in hypertensive and normotensive subjects.

OBJECTIVES AND PATIENTS: To determine whether or not the pulsatile component of blood pressure (BP) measured centrally and peripherally allows a separation between hypertensive and normal subjects, as well as within hypertensive and normal subjects, as well as within hypertensive patients. We tested the hypothesis that the difference in central and peripheral pulse pressures is increased in hypertensive, compared to normotensive persons, and that this component is influenced by genetic variance. We studied 46 hypertensive patients and 56 age-matched normal subjects, as well as 10 hypertensive families with 74 members of the same age range. DESIGN: Pulse pressure was measured at the brachial artery and the digital artery in the standing and supine position. The difference in the pulse pressures between these sites was calculated. Further, digital volume-pulse amplitude and stroke volume measurements were determined with impedance plethysmography. RESULTS: The differences between central and peripheral pulse pressures were similar in hypertensive patients compared to normal subjects, regardless of posture. However, in the standing position the frequency distribution of this variable in hypertensive patients was bimodal and split into two significantly different distributions (P < 0.05) with peaks at -24 mm Hg and -1 mm Hg, compared to a single peak at -11 mm Hg in normal subjects. Furthermore, these two subgroups of hypertensive patients differed in their brachial systolic BP (127 +/- 10 vs 134 +/- 12 mm Hg; P < 0.05), their brachial pulse pressures (32 +/- 8 vs 42 +/- 8 mm Hg; P < 0.05), and in their peripheral compliance (1.59 +/- 0.92 vs 2.21 +/- 1.00 microliter/mm Hg per 100 ml tissue; P < 0.05). The frequency distribution of pulse pressure differences was also bimodal in members of hypertensive families, even though most (46 out of 74) were normotensive. CONCLUSION: The difference between the digital and brachial pulsatile component may be a useful intermediary phenotype in essential hypertension. Furthermore, the nonuniform decreases in arterial compliance exhibited by our patients may be of pathogenic significance.

Management of immediate occlusion after carotid-reconstruction.

In spite of clear cut indications for surgery and the use of standardized techniques, operative failures are still to be found in carotid surgery, that is, the occurrence or aggravation of neurologic symptoms in connection with surgical reconstruction. These can present when the patient regains consciousness or develop from hours to days postoperatively. Within 3 1/2 years 93 carotid artery operations were performed in 79 patients. The purpose of our presentation is to describe 5 patients who developed neurologic deficits after a successful carotid reconstructive procedure within 18 hours to 10 days postoperatively. These patients were immediately returned to the operation room and reoperated upon. In 4 of 5 cases we achieved a complete recovery. Even though our patient content is small we want to point out that in patients who develop neurological deficits postoperatively after successful surgical procedures an immediate reoperation is mandatory, and may be completely successful. In almost all patients with recurrent neurological deficits after primarily successful reconstructions we carried out control angiograms. We want to emphasize that these procedures are not obligatory since the cause for reoperation is nearly always technical and will require operative revision.

Direct imaging of aligned neurofilament networks assembled using in situ dialysis in microchannels.

We report a technique to produce aligned neurofilament networks for direct imaging and diffraction studies using in situ dialysis in a microfluidic device. The alignment is achieved by assembling neurofilaments from protein subunits confined within microchannels. Resulting network structure was probed by polarized optical microscopy and atomic force microscopy, which confirmed a high degree of protein alignment inside the microchannels. This technique can be expanded to facilitate structural studies of a wide range of filamentous proteins and their hierarchical assemblies under varying assembly conditions.

Effect of sulfur availability on the integrity of amino acid biosynthesis in plants.

Amino acid levels in plants are regulated by a complex interplay of regulatory circuits at the level of enzyme activities and gene expression. Despite the diversity of precursors involved in amino acid biosynthesis as providing the carbon backbones, the amino groups and, for the amino acids methionine and cysteine, the sulfhydryl group and despite the involvement of amino acids as substrates in various downstream metabolic processes, the plant usually manages to provide relatively constant levels of all amino acids. Here we collate data on how amino acid homeostasis is shifted upon depletion of one of the major biosynthetic constituents, i.e., sulfur. Arabidopsis thaliana seedlings exposed to sulfate starvation respond with a set of adaptation processes to achieve a new balance of amino acid metabolism. First, metabolites containing reduced sulfur (cysteine, glutathione, S-adenosylmethionine) are reduced leading to a number of downstream effects. Second, the relative excess accumulation of N over S triggers processes to dump nitrogen in asparagine, glutamine and further N-rich compounds like ureides. Third, the depletion of glutathione affects the redox and stress response system of the glutathione-ascorbate cycle. Thus, biosynthesis of aromatic compounds is triggered to compensate for this loss, leading to an increased flux and accumulation of aromatic amino acids, especially tryptophan. Despite sulfate starvation, the homeostasis is kept, though shifted to a new state. This adaptation process keeps the plant viable even under an adverse nutritional status.

Expression analysis of a sucrose synthase gene from sugar beet (Beta vulgaris L.).

To investigate the expression pattern of sucrose synthase, a cDNA from tap roots of sugar beet (Beta vulgaris L.) was isolated using a heterologous sucrose synthase cDNA from potato. The 2762 bp long cDNA clone designated SBSS 1 encodes for a 822 amino acid polypeptide of a predicted molecular mass of 93.7 kDa. The deduced amino acid sequence of sugar beet sucrose synthase has homologies of 65-70% when compared to predicted amino acid sequences of sucrose synthases from other species. RNA blot analysis shows that SBSS1 is expressed most predominant in tap root under normal growth conditions. Cold treatment and anaerobiosis lead to an increase in the steady-state levels of SBSS 1 mRNA in leaf and root tissue. In tap root slices, sugars in various concentrations had no influence on the SBSS 1 transcript level. On the other hand, wounding resulted in a decreased transcript level.