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At least 217 records · Page 12Linked to original sources
PubMed2005

Using a miniaturized circuit and an asanguineous prime to reduce neutrophil-mediated organ dysfunction following infant cardiopulmonary bypass.

BACKGROUND: Contemporary infant cardiopulmonary bypass circuits require a blood prime. Blood, especially when stored, generates an inflammatory response, and may contribute to organ dysfunction following cardiopulmonary bypass. We determined whether using a miniaturized circuit and an asanguineous prime attenuated the post-bypass inflammatory response, and improved right ventricular and pulmonary function. METHODS: Sixteen infant piglets were placed into 3 groups based on prime components: group I (fresh blood), group II (stored blood), and group III (miniaturized circuit and asanguineous prime). Piglets were placed on cardiopulmonary bypass (100 mL.kg(-1).min(-1)), cooled to 18 degrees C, and underwent continuous perfusion (50 mL.kg(-1).min(-1)) for 30 minutes. They were rewarmed and separated from bypass. Serum tumor necrosis factor-alpha, right ventricular function, and pulmonary function were measured before and 30 minutes after bypass. Neutrophil priming activity in fresh and stored donor blood was also assessed. RESULTS: Animals in group III had significantly improved cardiopulmonary function than the groups receiving blood (right ventricular cardiac index [mL.kg(-1).min(-1)]: group I [18.8 +/- 4.8], group II [21.5 +/- 6.2], and group III [81.2 +/- 11.4], p < 0.001; and pulmonary vascular resistance index [dynes.mL(-1).kg(-1)]: group I [1169 +/- 409], group II [1610 +/- 486], and group III [214 +/- 63], p = 0.03). Tumor necrosis factor-alpha (pg.mL(-1)) was lower in group III (1465 +/- 39) than in the groups receiving blood (3940 +/- 777), p = 0.002. Neutrophil priming activity (nmol.min(-1)) was also higher in stored blood (3.7 +/- 6) than in fresh blood (1.9 +/- 0.2), p = 0.02. CONCLUSIONS: We have devised a unique miniaturized circuit that allows an asanguineous prime without hemodilution in an infant swine model. The employment of this circuit attenuates the post-bypass inflammatory response and has salutary effects on cardiopulmonary function.

Animals↗
PubMed2006

Miniature neurotransmission stabilizes synaptic function via tonic suppression of local dendritic protein synthesis.

Activity deprivation in neurons induces a slow compensatory scaling up of synaptic strength, reflecting a homeostatic mechanism for stabilizing neuronal activity. Prior studies have focused on the loss of action potential (AP) driven neurotransmission in synaptic homeostasis. Here, we show that the miniature synaptic transmission that persists during AP blockade profoundly shapes the time course and mechanism of homeostatic scaling. A brief blockade of NMDA receptor (NMDAR) mediated miniature synaptic events ("minis") rapidly scales up synaptic strength, over an order of magnitude faster than with AP blockade alone. The rapid scaling induced by NMDAR mini blockade is mediated by increased synaptic expression of surface GluR1 and the transient incorporation of Ca2+-permeable AMPA receptors at synapses; both of these changes are implemented locally within dendrites and require dendritic protein synthesis. These results indicate that NMDAR signaling during miniature synaptic transmission serves to stabilize synaptic function through active suppression of dendritic protein synthesis.

Action Potentials↗
PubMed2006

Miniature intravesical urethral lengthening procedure for treatment of pediatric neurogenic urinary incontinence.

PURPOSE: Resistance to flow in a fluid conduit is proportional to tube length divided by the radius to the fourth power (Poiseuille's law). We report the results of a miniature intravesical urethral lengthening procedure where outlet resistance is increased by minimizing the diameter of the intravesical urethral tube. MATERIALS AND METHODS: Nine pediatric patients with preoperative intractable incontinence underwent the miniature intravesical urethral lengthening procedure along with continent catheterizable stoma (9 patients) and bladder augmentation (8). The intravesical portion of the urethral lengthening was 3 cm (traditionally 6 cm), and the urethra was tubularized around an 8Fr feeding tube (traditionally a 16Fr catheter). After the tubularized caudal portion was tunneled under the trigone the cephalad part of the urethra was placed as an onlay to the posterior bladder wall without ureteral reimplantation. RESULTS: At a mean followup of 31 months (range 10 to 47) 8 patients reported dry intervals of 3 hours or more, with minor leak per urethra only if they were overdue on the catheterization schedule. Mean postoperative abdominal leak point pressure was 71 cm H(2)O (range 28 to 116). Upper tracts were well preserved in all patients. One patient required bladder neck closure for intractable incontinence. CONCLUSIONS: The miniature intravesical urethral lengthening procedure requires minimal bladder tissue and is easy to perform. It appears to be an effective alternative in bladder neck reconstructive techniques, avoiding the need for ureteral reimplantation due to its small size, while functioning as a pop-off valve when the bladder is overly full. This procedure should be avoided in patients who lack a trigonal bar.

Adolescent↗
PubMed1995

The rise times of miniature endplate currents suggest that acetylcholine may be released over a period of time.

Models of miniature endplate currents predict 20-80% rise times of 100 microseconds or less. These predictions are substantially less than most of the rise times recorded in the literature. New measurements were made of rise times at the frog neuromuscular junction using extracellular recording. The mean 20-80% rise time was 250 microseconds. Rise times were variable; at 20 degrees C, 95% of them fell in a range from 140 to 460 microseconds. The most questionable assumption in the models is that the acetylcholine (ACh) is released instantaneously. Modifying the model, so that ACh diffuses from the vesicle through a pore, lengthens the rise time to observed levels. It has been proposed that ACh is released from the vesicle in exchange for Na+. However, the rise times of miniature endplate currents recorded in solutions in which the Na+ is replaced by sucrose are in the normal range. The Q10 for the rise of miniature endplate currents is approximately 2, which is consistent with the models and with temperature effects on pore formation in mast cells.

Acetylcholine↗
PubMedDate not supplied

Progressive retinal atrophy in miniature longhaired dachshund dogs.

A form of generalized progressive retinal atrophy unlike other previously recorded canine retinal dystrophies has been investigated in Miniature Longhaired Dachshund dogs. Segregation patterns in litters from matings involving affected individuals were consistent with simple autosomal recessive inheritance. The earliest ophthalmoscopic signs, appearing at approximately 6 months of age and coinciding in some cases with the onset of nyctalopia, included changes in the granular appearance of the tapetal fundus followed by generalized tapetal hyper-reflectivity and retinal vascular attenuation; later there was irregular loss of pigment in the non-tapetal fundus and optic atrophy. However, there was marked variation in the age of onset and progression of the disease, even within a single litter. The electroretinogram was normal in waveform and latency in four affected littermates at 10 weeks of age but by 9 months was markedly reduced in amplitude in two pups and virtually extinguished in the others. Significant histological changes at 10.5 weeks of age included thinning of the outer nuclear layer, irregularity and attenuation of the rod photoreceptor outer segments and early disorganization of the rod outer segment disc lamellae. By 25 weeks the photoreceptors were grossly degenerate with short rounded inner segments and only residual amounts of outer segment material remaining. This condition in the Miniature Longhaired Dachshund is later in onset than rod-cone dysplasia in Irish Setters but significantly earlier than progressive retinal atrophy in Tibetan Terriers and progressive rod-cone degeneration in, for example, Miniature Poodles. The condition could therefore serve as a potentially useful model for retinitis pigmentosa in man.

Animals↗
PubMed2002

Miniature specimen shear punch test for UHMWPE used in total joint replacements.

Despite the critical role that shear is hypothesized to play in the damage modes that limit the performance of total hip and knee replacements, the shear behavior of ultra-high molecular weight polyethylene (UHMWPE) remains poorly understood, especially after oxidative degradation or radiation crosslinking. In the present study, we developed the miniature specimen (0.5 mm thickness x 6.4mm diameter) shear punch test to evaluate the shear behavior of UHMWPE used in total joint replacement components. We investigated the shear punch behavior of virgin and crosslinked stock materials, as well as of UHMWPE from tibial implants that were gamma-irradiated in air and shelf aged for up to 8.5 years. Finite element analysis, scanning electron microscopy, and interrupted testing were conducted to aid in the interpretation of the shear punch load-displacement curves. The shear punch load-displacement curves exhibited similar distinctive features. Following toe-in, the load-displacement curves were typically bilinear, and characterized by an initial stiffness, a transition load, a hardening stiffness, and a peak load. The finite element analysis established that the initial stiffness was proportional to the elastic modulus of the UHMWPE, and the transition load of the bilinear curve reflected the development of a plastically deforming zone traversing through the thickness of the sample. Based on our observations, we propose two interpretations of the peak load during the shear punch test: one theory is based on the initiation of crystalline plasticity, the other based on the transition from shear to tension during the tests. Due to the miniature specimen size, the shear punch test offers several potential advantages over bulk test methods, including the capability to directly measure shear behavior, and quite possibly infer ultimate uniaxial behavior as well, from shelf aged and retrieved UHMWPE components. Thus, the shear punch test represents an effective and complementary new tool in the armamentarium of miniature specimen mechanical testing methods for UHMWPE used in total joint replacement components.

Arthroplasty, Replacement↗
PubMed1984

Two types of miniature endplate potentials in Xenopus nerve-muscle cultures.

The frequency distribution of miniature endplate potential (mepp) amplitudes was examined in Xenopus nerve-muscle cultures. In the early days of nerve-muscle co-culture (1-2 days) the mepp amplitude histogram was skewed in the majority of muscle cells and had a peak approximately at 0.6 mV. Innervated muscle cells in older cultures (3-4 days), however, often had two peaks. The distribution at the smaller amplitude was skewed and the peak was approximately at 0.6 mV, whereas the peak at the larger amplitude was more-or-less symmetrical and about 18 times greater than the small one. The time-to-peak value of mepps in the peak at the smaller amplitude was slightly smaller than those in the peak at the larger amplitude. Small mepps were not due to electrotonic spread of mepps generated at distant sites nor due to focal development of acetylcholinesterase. Two peaks were also observed in the amplitude histogram of extracellularly recorded miniature endplate currents. Therefore, these two types of mepps must be generated practically at the same site. Mepps in the peak at the smaller amplitude had similar properties as sub-miniature endplate potentials (s-mepps) described in the adult animal. The second symmetrical peak in older cultures may correspond to classical mepps (c-mepps). The age-dependent shift from the amplitude distribution with one peak to that with two peaks has also been reported in developing and regenerating neuromuscular junctions. Therefore, I suggest that during development in culture, mepps appear at first as s-mepps and subsequently c-mepps develop, forming the second peak in the amplitude histogram.

Age Factors↗
PubMed2000

Vesicle number does not predict postsynaptic measures of miniature synaptic activity frequency in cultured cortical neurons.

We tested the hypothesis that heterogeneity in the frequency of miniature synaptic activity reflects differences in the number of vesicles present in presynaptic terminals. Using imaging techniques, we measured dendritic miniature synaptic calcium transients attributed to the spontaneous release of single transmitter quanta. Following imaging, the identified neurons were processed for serial transmission electron microscopy. At sites of quantal Ca(2+) transients mediated by N-methyl-D-aspartate receptors, we confirmed the presence of excitatory synapses and measured the total number of vesicles and the number of docked vesicles. We observed no correlation between the frequency of spontaneous miniature activity and either the total vesicle number or the number of docked vesicles. We conclude that the presynaptic vesicle complement as measured by ultrastructural analysis does not necessarily determine the frequency of spontaneous activity at synapses mediated by N-methyl-D-aspartate receptors.

Animals↗
PubMed2003

Cryosurgery in long bones with new miniature cryoprobe: an experimental in vivo study of the cryosurgical temperature field in sheep.

AIM: The aim of this in vivo study was to determine whether new miniature cryoprobes provide adequate tissue cooling in long bones by measuring the field of temperature under various conditions. METHODS: Freezings were performed in femoral and tibial bones of 10 sheep under general anaesthesia. Applying one cryoprobe, temperatures of -75 degrees C resp. -51 degrees C could be reached within 0.75 cm resp. 1.00 cm of the probe. RESULTS: Histological examinations revealed compact bone and marrow necroses along the isotherm in all 10 sheep. Using two cryoprobes simultaneously, a mean temperature decrease to -71 degrees C between the two cryoprobes was achieved thanks to the synergistic freezing effect. Looking at consecutive freezes, it was apparent that with similar cryoprobe end temperatures, the temperature dropped faster if the number of freezing cycles was increased. CONCLUSION: In conclusion, it was seen that despite its small diameter, the new miniature cryoprobe delivers adequate in vivo tissue cooling in long tubular bones. Employing the synergistic freezing effect by using two or more cryoprobes simultaneously, efficient in vivo freezing of larger bone segments is also possible. Thus, cryosurgery with the new miniature probes can provide a valuable complement to conventional resection of long tubular bones, and offers a viable alternative to surgical treatment of neoplastic diseases of the skeletal system.

Animals↗
PubMed1989

Histological evaluation of the effect of a miniature carbon dioxide laser on oral mucosa.

Carbon dioxide lasers emit infra-red radiation at a wavelength of 10600 nm, making them suitable for tissue vaporisation, incision and vessel coagulation. A miniature CO2 laser (length 24 cm), with an output of 4 Watts focused on to a spot of 0.2 mm diameter at a distance of 2.5 mm from the nozzle tip, has been developed. This hand-held, water-cooled device operates off a 12 V D.C. source and provides good access to most areas of the mouth. In order to evaluate the use of the miniature laser, a series of lesions were produced on the lateral margins of sheep's tongues, and histological examination of tissues carried out immediately after treatment and at 1 h; 1, 2, 4, 7, 14, 21, and 28 day intervals. Wounds showed clean cut margins with even removal of epithelium and superficial connective tissue. Damage appeared to be restricted to a 100 microns deep zone in the lamina propria. Epithelial regeneration had commenced at between 2 and 4 days, and re-covered the surface by 14 days. A residue of chronic inflammatory cells was present within the stroma at 21 days, but this had resolved by 28 days. The miniature carbon dioxide laser is effective in vaporising oral mucosa and also achieving haemostasis by vessel coagulation; its manoeuvrability and relatively low cost warrant further clinical evaluation for the treatment of mucosal lesions, such as leukoplakia.

Animals↗
PubMed1997

A miniaturized arrayed assay format for detecting small molecule-protein interactions in cells.

BACKGROUND: Two complementary approaches to studying the cellular function of proteins involve alteration of function either by mutating protein-encoding genes or by binding a small molecule to the protein. A mutagen can generate millions of genetic mutations; correspondingly, split-pool synthesis can generate millions of unique ligands attached to individual beads. Genetic screening of mutations is relatively straightforward but, in contrast, split-pool synthesis presents a challenge to current methods of screening for compounds that alter protein function. The methods used to screen natural products are not feasible for large libraries composed of covalently immobilized compounds on synthesis beads. The sheer number of compounds synthesized by split-pool synthesis, and the small quantity of individual compound attached to each bead require assay miniaturization for efficient screening. RESULTS: We present a miniaturized cell-based technique for the screening of ligands prepared by split-pool synthesis. Spatially defined droplets with uniform volumes of approximately 50-150 nanoliters (depending on well dimensions) are arrayed on plastic devices prepared using a combination of photolithography and polymer molding. Using this microtechnology, approximately 6,500 assays using either yeast cells or mammalian tissue culture can be performed within the dimensions of a standard 10 cm petri dish. We demonstrate that the biological effect of a small molecule prepared by split-pool synthesis can be detected in this format following its photorelease from a bead. CONCLUSIONS: The miniaturized format described here allows uniformly sized nanodroplets to be arrayed on plastic devices. The design is amenable to a large number of biological assays and the spatially arrayed format ensures uniform and controlled ligand concentrations and should facilitate automation of assays. The screening method presented here provides an efficient means of rapidly screening large numbers of ligands made by split-pool synthesis in both yeast and mammalian cells.

Animals↗
PubMed2001

Miniaturized HTS technologies - uHTS.

The transition from slow, manual, low-throughput screening to industrialized robotic ultra-high throughput screening (uHTS) in the past few years has made it possible to screen hundreds of thousands of chemical entities against a biological target in a short time-frame. The need to minimize the cost of screening has been addressed primarily by reducing the volume of sample to be screened. This, in turn, has resulted in the miniaturization of HTS technology as a whole. Miniaturization requires new technologies and strategies for compound handling, assay development, assay adaptation, liquid handling and automation in addition to refinement of the technologies used for detection systems and data management. This review summarizes current trends in the field of uHTS and illustrates the technological developments that are necessary to enable the routine application of miniaturized uHTS systems within an industrial environment.

Journal Article↗
PubMed2002

Analytical performance of a miniature cylindrical ion trap mass spectrometer.

The analytical performance of a fieldable cylindrical ion trap (CIT)-based miniature mass spectrometer is described. A detailed description of the instrument itself is to be found in the immediately preceding paper (Patterson, G. E.; Guymon, A. J.; Riter, L S.; Everly, M.; Griep-Raming, J.; Laughlin, B. C.; Ouyang, Z.; Cooks, R. G., Miniature Cylindrical Ion Trap Mass Spectrometer, Anal. Chem. 2002, 24, 6145-6153). Applications employing the MS/MS and MSn capabilities of the miniature instrument and analytical performance criteria are given here. The limit of detection for methyl salicylate, introduced as the pure vapor, is estimated as 1 pg. The resolution, R = m/delta m, where delta m, measured as full width at half-maximum, is estimated as 100. Monitoring of organic compounds in air is performed using a permeation membrane introduction device coupled to the mass spectrometer. Water monitoring is performed using an external membrane introduction mass spectrometry (MIMS) system, with acetophenone and toluene serving as model compounds. Data are given for chemical warfare agent simulants, methyl salicylate, and dimethyl methyl phosphonate (DMMP) in air. On-line detection of menthol vapor emitted from a cough drop is reported. Methyl salicylate in air gives a recognizable mass spectrum at 400 ppb in the ambient system, while use of a heated membrane brings the detection limit down to 10 ppb.

Journal Article↗
PubMed2003

Pulsed-ionization miniature ion mobility spectrometer.

We have demonstrated a miniature ion mobility spectrometer (IMS) that employs single pulses of corona discharge ionization. IMS spectra of both positive and negative ions generated from ambient air were measured as a function of drift field under various ionization conditions. Ion mobility spectra were studied with various pulse widths for both positive and negative ions, giving insights into mechanisms and kinetics of corona discharge ionization used in the miniature IMS. A combination of a pulsed potential with a steady dc bias was used to generate ions in the miniature IMS. There was a threshold dc potential for ion generation for a given pulse height. The dc ionization threshold was found to decrease linearly with increasing pulse height.

Journal Article↗
PubMed1998

A miniature analytical instrument for nucleic acids based on micromachined silicon reaction chambers.

In this paper, we describe a miniature analytical thermal cycling instrument (MATCI) to amplify and detect DNA via the polymerase chain reaction in real-time. The MATCI is an integrated, miniaturized analytical system that uses silicon-based, high-efficiency reaction chambers with integrated heaters and simple, inexpensive electronics to precisely control the reaction temperatures. Optical windows in the silicon and solid-state, diode-based detection components are employed to perform real-time fluorescence monitoring of product DNA production. The entire system fits into a briefcase and runs on rechargeable batteries. The applications of this miniaturized nucleic acid analysis system include clinical, research, environmental, and agricultural analyses as well as others which require rapid, portable, and accurate analysis of biological samples for nucleic acids. This paper describes the MATCI and presents results from ultrafast thermal cycling and real-time PCR detection. Examples include human genes and pathogenic viruses and bacteria.

Actins↗
PubMed2005

Relationship between folding and function in a sequence-specific miniature DNA-binding protein.

Previously, we have described a miniature protein-based approach to the design of molecules that bind DNA or protein surfaces with high affinity and specificity. In this approach, the small, well-folded protein avian pancreatic polypeptide acts as a scaffold to present and stabilize an alpha-helical or PPII-helical recognition epitope. The first miniature protein designed in this way, a molecule called p007, presents the alpha-helical recognition epitope found on the bZIP protein GCN4 and binds DNA with nanomolar affinity and exceptional specificity. In this work we use alanine-scanning mutagenesis to explore the contributions of 29 p007 residues to DNA affinity, specificity, and secondary structure. Virtually every residue within the p007 alpha-helix, and most residues within the p007 PPII helix, contribute to both DNA affinity and specificity. These residues include those introduced to make specific and nonspecific DNA contacts, as well as those that complete the miniature protein core. Moreover, there exists a direct correlation between the affinity of a p007 variant for specific DNA and the ability of that variant to select for specific DNA over nonspecific DNA. Although we observe no correlation between alpha-helicity and affinity, we observe a limited correlation between alpha-helicity and sequence specificity that emphasizes the role of coupled binding/folding in the function of p007. Our results imply that formation of a highly evolved set of protein.DNA contacts in the context of a well-packed hydrophobic core, and not the extent of intrinsic alpha-helical structure, is the primary determinant of p007 function.

Amino Acid Sequence↗
PubMed2003

Miniaturized chemical multiplexed sensor array.

Miniaturized tin oxide semiconductor sensors are fabricated directly by site-specific dip-pen nanopatterning using precursor inks derived from the sol-gel method. The good flow characteristics and strong affinity of the sols to measurement electrodes enable intimate contact. The measurable, reproducible, and proportionate changes in the resistance of the sensors when exposed to trace quantities of oxidative and reducing gases constitute the basis for such sensors. These sensors show rapid response and ultrafast recovery for the detection of nitrogen dioxide and acetic acid. Furthermore, an array of eight miniaturized sensors is created by doping the pristine tin-based sol ink with different metal ions; the different responses of each sensor to certain gases constitute a reference response spectrum that can be used to recognize the gas. Such recognition ability, instant response and rapid recovery, compact size, and integration with the current microelectronics platform make the miniaturized sensor array a significant development for the on-site and real-time detection of life-threatening gases.

Journal Article↗
PubMed1992

Rapid-time-course miniature and evoked excitatory currents at cerebellar synapses in situ.

Neurotransmission from mossy fibre terminals onto cerebellar granule cells is almost certainly mediated by L-glutamate. By taking advantage of the small soma size, limited number of processes and short dendrite length of granule cells, we have obtained high-resolution recordings of spontaneous miniature excitatory postsynaptic currents (m.e.p.s.cs) and evoked currents in thin cerebellar slices. Miniature currents have a similar time-course and pharmacology to evoked currents and consist of an exceptionally fast non-NMDA (N-methyl-D-aspartate) component (measured rise-time, 200 microseconds; estimated pre-filtered rise-time less than 100 microseconds; decay time constant, tau = 1.0 ms), followed by 50 pS NMDA channel openings that are directly resolvable. We could find no evidence for the recent proposal that miniature currents in granule cells are mediated solely by NMDA channels with a novel time course. The non-NMDA receptor component of m.e.p.s.cs has a skewed amplitude distribution, which suggests potential complications for quantal analysis. The difference in time course between the m.e.p.s.cs reported here and other synaptic currents in the brain could reflect differences in synaptic function or electrotonic filtering; the relative contribution of these possibilities has yet to be established.

Animals↗