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Hyperlipidemia, hyperlipemia, and hepatic lipidosis in American miniature horses: 23 cases (1990-1994).

The medical records of 23 American Miniature Horses with hyperlipidemia, hyperlipemia, or hepatic lipidosis were reviewed. The most common clinical signs were anorexia and lethargy. The mean duration of clinical signs was 2.4 days. A primary disease was identified in 19 cases. Enterocolitis was the most common primary disease (n = 10). Intentional feed restriction, as part of treatment for colic, resulted in hyperlipemia in 2 horses and hyperlipidemia in 1. Four horses had primary hyperlipemia, 3 of which had signs of hepatoencephalopathy secondary to hepatic lipidosis. Dextrose, heparin, and insulin were the most common treatments. The overall survival was 61% (14/23). All horses with peak serum triglyceride concentrations > 1,200 mg/dl died or were euthanatized, whereas all but 1 with peak serum triglyceride concentrations < 1,200 mg/dl survived. These findings suggest that when American Miniature Horses, like other ponies and donkeys, are in a negative energy balance, they can rapidly develop hyperlipidemia or hyperlipemia. Early detection and treatment may improve survival.

Animal Feed↗

Effects of tumor necrosis factor blockade on interleukin 6, lactate, thromboxane, and prostacyclin responses in miniature horses given endotoxin.

A monoclonal antibody (MAB) against equine tumor necrosis factor-alpha (Eq TNF) was used to investigate the role of TNF in cytokine, eicosanoid, and metabolic responses of Miniature Horses given endotoxin. Plasma concentrations of interleukin 6 (IL-6), lactate, thromboxane A2 metabolite, and prostacyclin metabolite (6-keto-PGF1 alpha) were measured in 10 Miniature Horses given 0.25 microgram of lipopolysaccharide (LPS; Escherichia coli O55:B5)/kg of body weight. Five horses were given Eq TNF MAB and 5 were given isotype-matched MAB as control. All horses were given 1.86 mg of antibody/kg by IV infusion, 5 minutes before LPS was given IV. Blood samples were taken 20 minutes before and at multiple intervals for 24 hours after LPS was given. Interleukin 6 bioactivity in plasma was measured, using IL-6-dependent cell line (B9). Eicosanoid activities were assessed by enzyme immunoassay, and plasma lactate concentration was determined enzymatically. Data were analyzed by ANOVA and Tukey's honest significant difference test for significant (P < 0.05) effect of treatment. Horses given Eq TNF MAB had significantly (P < 0.050) lower peak mean +/- SEM IL-6 (59 +/- 29 U/ml), lactate (16 +/- 2.00 mg/dl), and 6-keto-PGF1 alpha (254 +/- 79 pg/ml) values then did horses given control MAB (880 +/- 375 U/ml for IL-6; 26 +/- 0.04 mg/dl for lactate; and 985 +/- 290 pg/ml for 6-keto-PGF1 alpha). There was no effect of anti-TNF treatment on LPS-induced thromboxane A2 metabolite production. Tumor necrosis factor mediated IL-6, lactate, and prostacyclin responses, without affecting thromboxane production in horses given LPS.

6-Ketoprostaglandin F1 alpha↗

Implantable volume monitor and miniaturized stimulator dedicated to bladder control.

This paper describes a new miniaturized implantable bladder controller that is composed of 4 main parts: a volume monitoring device based on the tomography approach, a fully programmable miniaturized central processor and stimulator, a bidirectional data and power link, and an external controller. The proposed system is intended to restore both normal bladder functions (retention and voiding) to spinal cord injured patients. The system contains a mixed-signal (analog/digital) feedback loop to command the bladder functions through neuromuscular stimulation techniques. The implantable circuitry is powered by a single encoded radiofrequency carried and may have up to 8 independently controlled monopolar (4 bipolar) channels. The microstimulator is able to generate a wide range of stimulation patterns, including selective stimulation waveforms. In addition, an optical link transmits the state of the implant and volume monitoring results to the external controller.

Computer Simulation↗

Long term light-induced retinal degeneration in the miniature pig.

In developing a model of slow light-induced retinal degeneration, ten miniature pigs were submitted to constant lighting for a period ranging from one to three months. Post-lighting survival time ranged from zero to two months. Control and illuminated animals were examined for pupillary reflex, underwent fundus examination and an electroretinogram. After euthanasia, retinas were processed for histology with measure of outer nuclear layer thickness. All animals illuminated one or more months had pupillar reflex alteration. Mean outer nuclear thickness was 24.12 microns in the control and ranged from 18.36 to 21.45 microns in illuminated animals (mean reduction 20%). Despite the pigmentation of miniature pigs, consistent results were obtained in the absence of pharmacologic pupil dilation.

Animals↗

Stable properties of spontaneous EPSCs and miniature retinal EPSCs during the development of ON/OFF sublamination in the ferret lateral geniculate nucleus.

Retinal projections to the lateral geniculate nucleus (LGN) in ferrets progressively segregate into eye-specific laminae and subsequently into sublaminae that receive inputs from either ON-center or OFF-center afferents. To study the development of synaptic efficacy during a period of activity-dependent growth and reorganization in the CNS, we recorded spontaneous EPSCs (sEPSCs) from cells of the LGN during ON/OFF sublamination. We also examined retinal inputs specifically by stimulating the optic tract in the presence of strontium and recording evoked miniature EPSCs (emEPSCs). The rise times, areas, half-widths, and decay times of sEPSCs and emEPSCs and interevent intervals of sEPSCs recorded at the beginning of ON/OFF sublamination were not different from those recorded after its completion. Typically EPSC areas were small (10-20 fC) but varied greatly both within and between neurons. The frequency of sEPSCs was also quite variable, ranging from 0.2 to 5 Hz. sEPSCs were equivalent to miniature EPSCs recorded in the presence of tetrodotoxin, and both sEPSCs and emEPSCs were CNQX-sensitive. No difference was observed between sEPSCs recorded at room temperature and those recorded at 34 degreesC, and strontium could be substituted for calcium with no effect on sEPSC shape. These data argue for a remarkable stability in the components of at least AMPA-mediated synaptic transmission during a period of major synaptic rearrangement in the LGN.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

Effect of zolpidem on miniature IPSCs and occupancy of postsynaptic GABAA receptors in central synapses.

GABAA-mediated miniature IPSCs (mIPSCs) were recorded from layer V pyramidal neurons of the visual cortex using whole-cell patch-clamp recording in rat brain slices. At room temperature, the benzodiazepine site agonist zolpidem enhanced both the amplitude (to 138 +/- 26% of control value at 10 microM) and the duration (163 +/- 14%) of mIPSCs. The enhancement of mIPSC amplitude was not caused by an increase of the single-channel conductance of the postsynaptic receptors, as determined by peak-scaled non-stationary fluctuation analysis of mIPSCs. The effect of zolpidem on fast, synaptic-like (1 msec duration) applications of GABA to outside-out patches was also investigated. The EC50 for fast GABA applications was 310 microM. In patches, zolpidem enhanced the amplitude of currents elicited by subsaturating GABA applications (100-300 microM) but not by saturating applications (10 mM). The increase of mIPSC amplitude by zolpidem provides evidence that the GABAA receptors are not saturated during miniature synaptic transmission in the recorded cells. By comparing the facilitation induced by 1 microM zolpidem on outside-out patches and mIPSCs, we estimated the concentration of GABA seen by the postsynaptic GABAA receptors to be approximately 300 microM after single vesicle release. We have estimated a similar degree of receptor occupancy at room and physiological temperature. However, at 35 degreesC, zolpidem did not enhance the amplitude of mIPSCs or of subsaturating GABA applications on patches, implying that, in these neurons, zolpidem cannot be used to probe the degree of receptor occupancy at physiological temperature.

Algorithms↗

Surgical preparation of miniature swine for atherosclerosis research.

A 2-stage surgical procedure for chronic intestinal, vascular, and lymphatic sampling in miniature swine is described. Cannulas were placed in the jejunum and ileum, followed by a 2nd implantation of indwelling catheters in the portal and hepatic veins, caudal vena cava, abdominal aorta, and cisterna chyli. The techniques used proved that miniature swine can be surgically adapted to meet experimental requirements.

Animals↗

Fundus and histopathological study of radial optic neurotomy in the normal miniature pig eye.

OBJECTIVE: To demonstrate the fundus and histopathological changes in the normal miniature pig eye after radial optic neurotomy. METHODS: Ophthalmoscopic examinations, fundus photography, and fluorescein angiography were performed on 12 eyes that underwent radial optic neurotomy, 5 normal eyes, and 7 eyes that underwent vitrectomy (from 12 pigs in total) preoperatively and 1, 7, 14, 30, and 90 days postoperatively. These eyes were enucleated 120 days postoperatively. Sixteen eyes (of 8 pigs in total) that underwent radial optic neurotomy were enucleated 1, 3, 7, and 48 days (2 pigs each time) postoperatively. The enucleated eye sections were stained with hematoxylin-eosin, Masson trichrome, and Luxol fast blue. RESULTS: The retina radial to the site of radial optic neurotomy darkened gradually with the increasing curvature of the major retinal arteries. The filling time intervals from the retinal artery to the retinal vein were prolonged. At the incision site, there was a loss of nerve fibers, which were subsequently replaced by collagenous tissue. No anastomotic vessels formed by the end of the study. CONCLUSIONS: Postoperatively, the retinal circulation seemed somewhat sluggish compared with that seen preoperatively. Segmental retinal nerve atrophy eventually formed. The procedure itself may not be the sole factor for the formation of shunt vessels. CLINICAL RELEVANCE: To date, radial optic neurotomy is controversial and persuasive animal studies are lacking. In combining fluorescein angiography with histopathological examination, this study may be somewhat helpful.

Animals↗

Ontogeny of histochemical fiber types and muscle function in the masseter muscle of miniature swine.

In this study of masticatory maturation, the ontogeny of the histochemical fiber type composition of musculus masseter is examined in the omnivorous miniature swine (Sus scrofa). Fiber type characteristics are interpreted by comparison with electromyography (EMG) recorded during feeding behavior. Similar to locomotion studies, the results suggest a correspondence between the composition and arrangement of motor units and their recruitment pattern. Serial sections of masseter muscles from 10 minipigs, ranging from 2 weeks to slightly over 1 year of age, were stained for myosin adenosine triphosphatase (mATPase) activity to distinguish slow-twitch from fast-twitch fibers, and for nicotinamide adenosine dehydrogenase-tetrazolium reductase to assess the aerobic capacity of the same fibers. Although maintaining a uniformly high aerobic capacity throughout ontogeny and in adult animals, a transition is observed in the relative proportions of fast- and slow-twitch fibers. The primarily fast-twitch neonatal pig masseter eventually comprises approximately 25-30% slow-twitch fibers in adults, with a higher predominance of slow fibers in the deep (vs. superficial) and anterior (vs. posterior) regions of the muscle. Furthermore, while individual fibers of adult masseters generally stain for either alkaline- or acid-stable mATPase activity, a substantial proportion of cells in developing animals exhibits the presence of both isozymes. EMG results indicate functional heterogeneity within the masseter of adult pigs. During chewing, when pig chow is replaced by cracked corn, EMG activity in the deep portion of the muscle either decreases or increases slightly. In the superficial portion, however, muscle amplitudes become dramatically higher for corn, surpassing levels generated for chewing the less obdurate chow. These results are consistent with a behavioral transition from neonatal suckling to sustained mastication of foods of more complex textures eaten by adult pigs. The relationship between these fiber type and EMG results for pig masseter corresponds to those pertaining to motor unit recruitment in the extensor muscles of locomotion. Implications of this work for the evolutionary morphology of mastication also are discussed.

Animals↗

Proteoglycan and proteome profiling of central human pulmonary fibrotic tissue utilizing miniaturized sample preparation: a feasibility study.

The objective of this study was to isolate fibrotic cells from human lung biopsies taken from different central pulmonary locations. A comparison was made of cell morphology, proteoglycan- and protein-expression in mesenchymal cell cultures obtained from human bronchial biopsies from patients with asthmatic-like disorders. We isolated viable cells from 10 out of the 12 biopsies. The fibroblast-like cells were positive for the biomarker a-smooth muscle actin, indicating that the cells were in an activated state. Two different types of fibroblast-like cells were observed from human pulmonary connective tissue; one of contractile type with lamellipodia that facilitate migration and a second cell type with an increased cell size, which most likely is of a synthetic phenotype. This is the first evidence of alterations in the proteoglycan expression pattern of versican, perlecan, biglycan and decorin which can be linked to the pathophysiological state of asthmatics proven by a combination of solid-phase extraction by reversed phase and by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Protein expression analysis using two-dimensional electrophoresis was interfaced to miniaturized sample preparation techniques using microcapillary extraction. Four protein groups were identified; cytoskeletal, adhesion, scavenger and metabolic proteins. These patient's proteomes showed a high degree of heterogeneity between patients but larger homogeneity within biopsies derived from different locations of the same patient.

Actins↗

Temporomandibular joint in miniature pigs: anatomy, cell replication, and relation to loading.

The mechanical environment is a regulator of growth and adaptation of the musculoskeletal system, including joints. Although pigs (Sus scrofa) are used frequently as models for temporomandibular joint (TMJ) dysfunction, no systematic description of microanatomy exists for this species. We injected the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) into 10- to 11-month-old miniature pigs that were undergoing measurements of TMJ bone strain. Ten hr later, the animals were sacrificed and their heads were perfused. Histological sections were used to map the distribution of replicating cells. Additional observations were made on gross dissections of jaw joints obtained from an abattoir. The pig TMJ is better supported than that of humans laterally and medially, but more vulnerable posteriorly. The posterior attachment area of the intra-articular disc is fibro-fatty rather than vascular, as in humans. Cartilage lines the articular eminence as well as the condylar surface. At the posterosuperior region of the condyle, the cartilage ends abruptly and is replaced by an invaginating, actively replicating periosteum. Almost all of the BrdU-labeled cells resided in the prechondroblastic zones. The condyle had more replicating cells than did the eminence (P < 0.02), but lateral and medial locations did not differ in either element. In sagittal sections, the condyle had more replicating cells posteriorly (P < 0.001), but no A-P differences were seen in the eminence. Comparisons of these data with data on bone strain indicate that increased loading is negatively associated with cell replication.

Animals↗

Morphology of the enamel organ in the miniature swine.

In recent years, the dentition of the pig has been increasingly used as a model for the study of amelogenesis. Indeed, much of our current knowledge on enamel formation derives from biochemical and physicochemical analyses of the organic and inorganic components, respectively, of porcine enamel. As an extension of this previous work, and as the first step in our attempt to correlate known enamel matrix and mineral changes with adjacent enamel organ morphology, the present study was undertaken to provide a description of the morphological events occurring in the enamel organ during porcine amelogenesis. Two-week-old miniature swine (minipigs) were fixed by vascular perfusion with glutaraldehyde, the deciduous teeth present at this age were embedded in Epon resin and sectioned, and the cells of the enamel organ at each of the various developmental stages of amelogenesis were examined by light and transmission electron microscopy. In many respects, the morphology of the porcine enamel organ was similar to that previously described in other mammalian species. On the other hand, several particularities were noted and these are discussed in the context of available data correlating cell ultrastructure with putative function during enamel formation.

Ameloblasts↗

Miniaturized pressure-guide-wire: evaluation in vitro and in isolated hearts.

UNLABELLED: The present study focused on the accuracy and reproducability of measurements performed with a miniaturized pressure guidewire system (PGWS) under standardized in vitro conditions and in an isolated pig heart model. We used a pressure guide wire system provided by Pressure Guide, Radi Medical Systems, Uppsala, Sweden. Pressures were recorded by a fiber optic pressure sensor located laterally and 3 cm from the distal end. The basic principle is that the element modulates an optical reflection by pressure induced elastic movements. The light source is an emitting diode in the control unit. The PGWS replaces a standard 0.018 in. guidewire. The PGWS was introduced through a Y-adapter in a recirculating perfusion model, consisting of tubing with internal diameters from 1.00 to 4.00 mm. Different perfusion pressures (range 20-140 mm Hg) were generated by a pressure controlled roller pump. RESULTS: Different lumen sizes from 1 to 4 mm had no influence on the measurements. a) In a range from 20 to 140 mm Hg no significant differences were found between the perfusion pressure and the pressures obtained by the PGWS, resulting in an r-value of 0.99 and an equation of y = 1.04x + 1.9. b) In pressure measurements proximal and distal of an artificial stenoses resulting in gradients from 0 to 100 mm Hg, the PGWS correctly identified the pressure difference with an equally high accuracy, 1, 2, and 3 cm behind the stenoses.(ABSTRACT TRUNCATED AT 250 WORDS)

Angioplasty, Balloon, Coronary↗

Grapefruit pectin inhibits hypercholesterolemia and atherosclerosis in miniature swine.

We studied the effect of dietary grapefruit pectin on plasma cholesterol and the development of atherosclerosis in 18 miniature swine. Pigs were randomized to one of three diets: no added fat (I), added fat/cellulose (II), and added fat/pectin (III). Plasma cholesterol was measured monthly. Arteries were examined for atherosclerosis at the termination of the experiment. Pectin supplementation of an added fat diet resulted in a significantly lower average plasma cholesterol than did cellulose supplementation (168 mg/dl vs. 249 mg/dl, p less than 0.05). The pectin-fed pigs also developed less atherosclerosis of their aortas (1.1% vs. 7.0%, p less than 0.05) and coronary arteries (2.9% vs. 26.2% cross-sectional narrowing, p less than 0.05). Plasma cholesterol levels correlated with the severity of aortic (r = 0.836) and coronary artery (r = 0.735) atherosclerosis. We conclude that dietary grapefruit pectin supplementation inhibits hypercholesterolemia and appears to be proportionately protective against atherosclerosis.

Animals↗

A rapid and efficient method for the screening of acid phosphatase 1 in young tomato seedlings, and for the identification of root-knot nematode species using miniaturized polyacrylamide gel electrophoresis.

A relatively rapid and highly sensitive miniaturized polyacrylamide gel electrophoresis technique is described for the analysis of certain isozymes from single cotyledons of tomato seedlings and from single females of the root-know nematode (Meloidogyne spp.). Homogenates from single tomato cotyledons (7, 14, 21, and 28 days old) were electrophoresed and stained for acid phosphatase 1 (Aps 1) activity. Cotyledons from plants of all the above age groups showed good Aps 1 activity. Nondestructive screening for tomato Aps 1 is therefore feasible, using very small samples, from as young as 7-day-old tomato seedlings. This could be of important use in expediting root-knot nematode resistance (based on the Aps 1-linked resistance gene Mi) screening for breeding programs, or F1 testing for seed production purposes. In addition, the mini-polyacrylamide gel electrophoresis technique was useful for determination of the Aps 1 allelic contribution to the total enzyme activity. The system was also used to detect malate dehydrogenase and esterase isozyme activity from single adult females of the four common root-knot nematodes, Meloidogyne arenaria, M. hapla, M. incognita, and M. javanica, with equally good results, enabling species discrimination.

Acid Phosphatase↗

Miniaturized movable contactless conductivity detection cell for capillary electrophoresis.

A miniaturized capacitively coupled contactless conductivity detector (mini-C(4)D) cell has been designed which is small enough to allow it to slide along the effective capillary length inside the capillary cassette of an Agilent capiillary electrophoresis system (CE) (or other CE brand of similar construction), including the possibility of positioning it close to the point of optical detection (4 cm), or even putting two such detector cells in one cassette. The cell was tested and the performance characteristics (noise, sensitivity, and peak width) were compared with those obtained with the previously used large C(4)D cell. No significant differences were observed. The mini-C(4)D was used in simultaneous separations of common cations and anions where its advantage over a larger C(4)D cell is the ability to vary the point of detection with the mini-C(4)D cell continuously at any point along the capillary length, so that the optimum apparent selectivity can be chosen. Other applications include providing a convenient second point of detection in addition to photometric detection, such as to measure accurately the linear velocity of a zone, or to allow placement of two mini-C(4)D cells in one capillary cassette simultaneously.

Electrochemistry↗

Exploiting the benefits of miniaturization for the enhancement of DNA microarrays.

The present study demonstrates that the best way to enhance DNA microarray assays, both in terms of analysis speed and in final spot intensity, is to dissolve the available molar amount of sample in the smallest possible buffer volume and to subsequently convect this solution continuously across the surface of the array. The presently proposed shear-driven flow system is pre-eminently suited for this task, as it allows to induce strongly enhanced lateral transport rates, independently of the degree of miniaturization of the hybridization chamber. This transport enhancement method, however, only increases the hybridization rate and not the final spot intensity, as neither can any of the other transport enhancement methods already proposed in literature. A series of experiments with synthetic single-stranded (ssDNA) samples and an accompanying mass balance analysis are presented to demonstrate these points.

DNA, Single-Stranded↗