Automated,Ultra‐Fast Laser‐Drilling of Nanometer Scale Pores and Nanopore Arrays in Aqueous Solutions

The ability to quickly and reliably fabricate nanoscale pore arrays in ultra‐thin membranes such as silicon nitride (Si_xN) is extremely important for the growing field of nanopore biosensing. Laser‐based etching of thin Si_xN membranes immersed in aqueous solutions has recently been demonstrated as a method to produce stable functional pores. Herein, the principal mechanism governing material etching and pore formation using light is investigated. It is found that the process is extremely sensitive to the relative content of Si over N atoms in the amorphous membrane, produced by chemical vapor deposition. Commonly, Si_xN membranes are made to be Si‐rich to increase their mechanical stability, which substantially reduces the material's bandgap and increases the density of Si‐dangling bonds. Hence, even minimal batch‐to‐batch variation may lead to remarkably different etch rates. It is shown that higher Si content results in orders of magnitude faster etching rates. This rate is further accelerated in an alkaline environment allowing on‐demand controlled nanopore formation in about 10 s time even at low laser radiation intensities. These results highlight that photoactivation of the Si_xN by the incident beam is critical to the chemical etching process and can be used to readily produce nanopore arrays at any specific location.     

DNA nanomechanical switches under folding kinetics control

Existing DNA nanodevices operate at equilibrium under changes in solution composition. We propose an alternative DNA switch design that can be driven and maintained out of equlibrium, under fixed chemical conditions. Moderate cooling rate after heat denaturation drives the switch to its lowest energy conformation, while rapid cooling (>100 degrees C/ms) locks the molecule in a unique alternative conformation that is retained over weeks at room temperature. This reversible process is probed using fluorescent energy transfer. DNA switches operating out of equilibrium should be more amenable to nanotechnology applications and scalable integration.     

Increased benzodiazepine receptor density in the prefrontal cortex in patients with panic disorder

There are published studies concerning a regionally changed function of GABA-benzodiazepine-receptor-complexes in anxiety disorder. These studies implicate the limbic lobe, the brainstem and the prefrontal cortex. Using 123I-Iomazenil and single photon emission tomography (SPET) we investigated the benzodiazepine receptor density in twelve patients with panic disorder who had never been treated with benzodiazepines before. Nine age- and sex-matched volunteers who were free of mental illness served as controls. Patients with panic disorder showed a significant increase of benzodiazepine receptor binding in the right supraorbital cortex and a trend to an increased uptake in the right temporal cortex. There was no correlation between receptor density and scores on Spielberger's State Trait Anxiety Inventory in the patient group. Since the findings cannot be explained by benzodiazepine exposure we hypothesize an upregulation due to functional or neuroanatomic changes (at least) in the frontotemporal cortex.     

Pex17p of Saccharomyces cerevisiae is a novel peroxin and component of the peroxisomal protein translocation machinery

The function of the human papillomavirus type 11 (HPV 11) E1--E4 spliced protein is not known. E1--E4 protein in HPV-infected tissue is detected in the cytoplasm of differentiated epithelial cells and as immunoreactive bands corresponding to potential monomers, dimers and trimers in immunoblots. The yeast two-hybrid system was employed to test for self association of the HPV 11 E1--E4 protein. To confirm the results of the yeast two-hybrid experiments, coimmunofluorescence studies of a green fluorescent fusion protein (GFP-E1--E4) and a T7 epitope-tagged E1--E4 protein were performed in C33a keratinocytes. E1--E4 protein was shown to self associate in the yeast two-hybrid system, and this result was confirmed by colocalization of GFP-E1--E4 and T7-E1(wedge)E4 proteins in keratinocytes. Analysis of E1--E4 mutants established that the C-terminus was required for self association and that sequences in the N-terminus influenced the intracellular localization of E1--E4 protein. The intracellular expression patterns of GFP-E1--E4 and GFP-E1--E4 mutants were correlated with E1--E4 binding in the yeast two-hybrid system. Those E1--E4 mutants that did not self associate in the yeast two-hybrid system were detected as diffuse cellular fluorescence when expressed as GFP fusions. In contrast, GFP-E1--E4 was detected as a perinuclear aggregate. All E1--E4 mutants capable of associating with E1--E4 in the yeast two-hybrid system were detected as aggregates when expressed as GFP fusion proteins in keratinocytes.     

The effect of epidural injection of betamethasone or bupivacaine in a rat model of lumbar radiculopathy

STUDY DESIGN: The effect of epidural injection of betamethasone or bupivacaine was investigated in an animal model of lumbar radiculopathy. OBJECTIVE: To investigate the effects of an epidural steroid (betamethasone) or a local anesthetic (bupivacaine) in an animal model of radiculopathy produced by nerve root irritation. SUMMARY OF BACKGROUND DATA: Epidural injections are commonly used for the treatment of low back pain and sciatica. However, efficacy remains controversial, and there is a paucity of basic information to support clinical use or the injections. METHODS: Fifty-one rats were used. The left L4 and L5 nerve roots were loosely ligated with chromic gut, and either betamethasone, bupivacaine, betamethasone in combination with bupivacaine, or saline was injected using an epidurally placed catheter. The effects of epidural injection were evaluated using response to noxious stimuli and immunohistochemical methods. RESULTS: In betamethasone-treated rats (either alone or in combination with bupivacaine), thermal hyperalgesia was significantly less (P < 0.010 after surgery than that in saline- or bupivacaine-treated groups, in which the hyperalgesia was maximum at 2-3 postoperative weeks before resolving 5 weeks after surgery. Immunohistochemical analysis did not correlate with these results. CONCLUSIONS: Epidural steroid injection has a significant effect on the thermal hyperalgesia produced in a model of radiculopathy, which may provide clinical support for advocates of epidural steroids.     

Axonal and dendritic transport in Purkinje cells of cerebellar slice cultures studied by microinjection of horseradish peroxidase

BACKGROUND: The pathophysiological mechanisms in non-ulcer dyspepsia are incompletely understood. AIMS: To compare gastric motor and sensory functions in Helicobacter pylori positive or negative patients with non-ulcer dyspepsia. PATIENTS: Seventeen patients with non-ulcer dyspepsia and 16 asymptomatic controls. METHODS: The following were evaluated: gastrointestinal symptoms; gastric emptying and orocaecal transit of solids; abdominal vagal function; gastric compliance; fasting and postprandial gastric tone and phasic contractions; symptoms during ingestion of cold water and during the distension of an intragastric bag; and somatic sensitivity and personality profile (Minnesota Multiphasic Personality Inventory, MMPI). RESULTS: Gastric accommodation was reduced in H pylori negative dyspeptics relative to controls; the degree of accommodation was unrelated to H pylori status in dyspeptics. Increased postprandial gastric sensation was more frequent among H pylori positive patients (4/5 H pylori positive versus 4/12 H pylori negative patients). Intragastric meal distribution and orocaecal transit were normal; gastric emptying at four hours was abnormal in 4/17 patients. Vagal dysfunction was rare. Eight of 17 patients had somatisation or depression on MMPI. CONCLUSION: Impaired gastric accommodation is frequent in non-ulcer dyspepsia and seems to be unrelated to vagal efferent dysfunction. H pylori infection does not seem to influence gastric accommodation, but is associated with heightened sensitivity in dyspeptics. Therapeutic approaches that restore normal postprandial accommodation and gastric sensitivity should be tested in non-ulcer dyspepsia.     

Immunohistochemical studies on neurofilamentous hypertrophy in degenerating retinal terminals of the olivary pretectal nucleus in the rat

Following section of the optic nerve, degenerating retinal terminals reveal an accumulation of neurofilaments (neurofilamentous hypertrophy) as demonstrated by silver impregnation techniques or electron microscopy. The present study examined degenerating retinal terminals by means of immunohistochemistry and antibodies specific for the triplet of neurofilament proteins of low (NF-L), medium (NF-M), and high (NF-H) molecular weight class. Following unilateral optic nerve section in the rat and survival of 1, 2, 4, 8, and 21 days, brains were perfused with aldehyde fixative, sliced on a vibratome and stained for neurofilaments by using the peroxidase-antiperoxidase technique. Other brains were frozen, cut in the native state, and slide-mounted sections were fixed by acetone. Side comparisons in visual pathways were made in frontal sections, taking advantage of the near complete crossing of retinal fibers in the rat. Anterograde degeneration of axons occurred in the optic tract and branchium colliculi. Changes of terminals were investigated in the olivary pretectal nucleus, which contains a dense aggregation of retinal terminals in the core region. The optic tract and branchium colliculi showed a reduction in immunostaining for neurofilament proteins following axotomy. Within the core region of the olivary pretectal nucleus, strong increases of immunoreactivity of NF-L and NF-M were detected beginning at 2 days postlesion and persisting at 8 days. No changes in NF-H proteins were found in the terminal regions with three different antibody probes. The increase in immunostaining reflects the accumulation of neurofilament proteins in the degenerating retinal terminals, i.e., neurofilamentous hypertrophy.(ABSTRACT TRUNCATED AT 250 WORDS)     

Single-plant sourcing decisions with a multidimensional knapsack model

This paper addresses sourcing decisions and how those decisions can affect the management of a company’s assets. The study begins with a single-plant insourcing problem in which the facility chooses, from a list of parts, which parts to bring in-house. This model is extended to include an outsourcing element where, in addition to the decision of which parts to insource, parts currently in-house are considered for outsourcing. The selection is based on maximizing the value of the selected parts while remaining within the plant’s capacity. This is defined as the sourcing problem and modeled as a multidimensional knapsack problem (MKP). Characteristics of the sourcing problem are identified and compared to standard MKP test problems, then utilized to provide guidelines for generating industry-motivated data sets. Experiences with modeling and solving the sourcing problem for a sheet metal fabrication facility are reported.     

A space allocation algorithm for assembly line components

A component stockout during the assembly process is one of the most undesirable events that can occur since the resulting line stoppage is associated with extremely high costs. In this paper we address the problem of allocating space to components along the line, subject to practical constraints. The objective is to maximize the line fill-rate; namely, the probability of no line stoppages due to a lack of components between consecutive replenishments. A model for calculating the line fill-rate is presented. This model is then incorporated into a design algorithm that determines the space allocation. A large experiment for moderate-sized and large-scale problems is performed to evaluate the algorithm performance. Our experiment indicates this performance to be excellent, producing optimal solutions on moderate-sized problems in most cases and solutions that are better than three intuitive rules for most of the large-scale problems considered.     

Potential roles of nitrate and live yeast culture in suppressing methane emission and influencing ruminal fermentation,digestibility, and milk production in lactating Jersey cows

Concern over the carbon footprint of the dairy industry has led to various dietary approaches to mitigate enteric CH₄ production. One approach is feeding the electron acceptor NO₃^?, thus outcompeting methanogens for aqueous H₂. We hypothesized that a live yeast culture (LYC; Saccharomyces cerevisiae from Yea-Sacc 1026, Alltech Inc., Nicholasville, KY) would stimulate the complete reduction of NO₃^? to NH₃ by selenomonads, thus decreasing the quantity of CH₄ emissions per unit of energy-corrected milk production while decreasing blood methemoglobin concentration resulting from the absorbed intermediate, NO₂^?. Twelve lactating Jersey cows (8 multiparous and noncannulated; 4 primiparous and ruminally cannulated) were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Cattle were fed diets containing 1.5% NO₃^? (from calcium ammonium nitrate) or an isonitrogenous control diet (containing additional urea) and given a top-dress of ground corn without or with LYC, with the fourth week used for data collection. Noncannulated cows were spot measured for CH₄ emission by mouth using GreenFeed (C-Lock Inc., Rapid City, SD). The main effect of NO₃^? decreased CH₄ by 17% but decreased dry matter intake by 10% (from 19.8 to 17.8 kg/d) such that CH₄:dry matter intake numerically decreased by 8% and CH₄:milk net energy for lactation production was unaffected by treatment. Milk and milk fat production were not affected, but NO₃^? decreased milk protein from 758 to 689 g/d. Ruminal pH decreased more sharply after feeding for cows fed diets without NO₃^?. Acetate:propionate was greater for cows fed NO₃^?, particularly when combined with LYC (interaction effect). Blood methemoglobin was higher for cattle fed NO₃^? than for those fed the control diet but was low for both treatments (1.5 vs. 0.5%, respectively; only one measurement exceeded 5%), indicating minimal risk for NO₂^? accumulation at our feeding level of NO₃^?. Although neither apparent organic matter nor neutral detergent fiber digestibilities were affected, apparent N digestibility had an interaction for NO₃^? × LYC such that apparent N digestibility was numerically lowest for diets containing both NO₃^? and LYC compared with the other 3 diets. Under the conditions of this study, NO₃^? mitigated ruminal methanogenesis but also depressed dry matter intake and milk protein yield. Based on the fact that few interactions were detected, LYC had a minimal role in attenuating negative cow responses to NO₃^? supplementation.