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PubMed2003

The use of a new miniature cryoprobe for ablation of bone tissue: in vivo assessment of the probe and application of the method to bone in a sheep model.

BACKGROUND: So far, modern miniature cryoprobes were used for local destruction of soft tissue tumours without damaging the adjacent healthy tissue. In this study, cryoablation methodology was applied to bone and the cooling capacity of the probe was examined in vitro and in vivo. METHOD: Freezing was performed by cooling one or two probes (diameter 3.2 mm) to -180 degrees C with liquid nitrogen. The cooling capacity of the probes was determined optically and thermally against a homogeneous reference gel, followed by in vivo measurements on femoral and tibial sheep bone followed by histological examination. RESULTS: Thanks to the synergistic effect, the simultaneous use of 2 probes produced an almost spherical expansion of cold in the homogenous gelatin. During the in vivo freezes, the temperature curves showed a more moderate trend. Nevertheless, due to the synergistic effect, temperatures below -50 degrees C could be reached at a distance of 1 cm from the probe. No local or systemic intraoperative complications were observed. Histological examination revealed cell necrosis up into the -10 degrees C isotherm. CONCLUSIONS: Adequate tissue cooling of the bone matrix can be achieved with in vivo freezes by means of one or more miniature cryoprobes. Therefore, this probe could provide an alternative to or supplement surgical resection of pathological bone processes.

Animals↗

PubMed2006

Miniature bioreactors: current practices and future opportunities.

This review focuses on the emerging field of miniature bioreactors (MBRs), and examines the way in which they are used to speed up many areas of bioprocessing. MBRs aim to achieve this acceleration as a result of their inherent high-throughput capability, which results from their ability to perform many cell cultivations in parallel. There are several applications for MBRs, ranging from media development and strain improvement to process optimisation. The potential of MBRs for use in these applications will be explained in detail in this review. MBRs are currently based on several existing bioreactor platforms such as shaken devices, stirred-tank reactors and bubble columns. This review will present the advantages and disadvantages of each design together with an appraisal of prototype and commercialised devices developed for parallel operation. Finally we will discuss how MBRs can be used in conjunction with automated robotic systems and other miniature process units to deliver a fully-integrated, high-throughput (HT) solution for cell cultivation process development.

Journal Article↗

PubMed2024

Adaptive or non-adaptive? Cranial evolution in a radiation of miniaturized day geckos.

Lygodactylus geckos represent a well-documented radiation of miniaturized lizards with diverse life-history traits that are widely distributed in Africa, Madagascar, and South America. The group has diversified into numerous species with high levels of morphological similarity. The evolutionary processes underlying such diversification remain enigmatic, because species live in different ecological biomes, ecoregions and microhabitats, while suggesting strikingly high levels of homoplasy. To underscore this evolutionary pattern, here we explore the shape variation of skull elements (i.e., cranium, jaw and inner ear) using 3D geometric morphometrics and phylogenetic comparative methods on computed tomography scans (CT-scan) of a sample encompassing almost all recognized taxa within Lygodactylus. The results of this work show that skull and inner ear shape variation is low (i.e., there is high overlapping on the morphospace) across geographic regions, macrohabitats and lifestyles, implying extensive homoplasy. Furthermore, we also found a strong influence of allometry shaping cranial variation both at intra and interspecific levels, suggesting a major constraint underlying skull architecture, probably as a consequence of its miniaturization. The remaining variation that is not allometric is independent of phylogeny and ecological adaptation and can probably be interpreted as the result of intrinsic developmental plasticity. This, in turn, supports the interpretation that speciation in this group is largely concordant with a non-adaptive hypothesis, which results mainly from vicariant processes.

Animals↗

PubMed2005

The miniature pig as an animal model in biomedical research.

Crucial prerequisites for the development of safe preclinical protocols in biomedical research are suitable animal models that would allow for human-related validation of valuable research information gathered from experimentation with lower mammals. In this sense, the miniature pig, sharing many physiological similarities with humans, offers several breeding and handling advantages (when compared to non-human primates), making it an optimal species for preclinical experimentation. The present review offers several examples taken from current research in the hope of convincing the reader that the porcine animal model has gained massively in importance in biomedical research during the last few years. The adduced examples are taken from the following fields of investigation: (a) the physiology of reproduction, where pig oocytes are being used to study chromosomal abnormalities (aneuploidy) in the adult human oocyte; (b) the generation of suitable organs for xenotransplantation using transgene expression in pig tissues; (c) the skin physiology and the treatment of skin defects using cell therapy-based approaches that take advantage of similarities between pig and human epidermis; and (d) neurotransplantation using porcine neural stem cells grafted into inbred miniature pigs as an alternative model to non-human primates xenografted with human cells.

Aneuploidy↗

PubMed2003

Drosophila miniature and dusky encode ZP proteins required for cytoskeletal reorganisation during wing morphogenesis.

We have characterised the function of two Drosophila genes, miniature and dusky, that are required for the morphological reorganisation of the apical membrane during wing epidermis differentiation. These genes encode transmembrane proteins containing a ZP (zona pellucida) domain and are homologous to several vertebrate and invertebrate apical matrix components. miniature and dusky are only expressed in tissues secreting a cuticle, and the Min protein localises to the apical membrane during the early stages of cuticle formation. We propose that Min and Dusky form a novel subfamily within the ZP domain proteins and are specifically involved in the interactions between the apical membrane, the cytoskeleton and the forming cuticle.

Amino Acid Sequence↗

PubMed2004

Passive immunoneutralization of endogenous inhibin increases ovulation rate in miniature Shiba goats.

We reported previously that passive immunization against inhibin enhances follicular growth and increases the ovulation rate. However, the ovulation rate was not comparable to the number of follicles. Therefore, the aim of this study was to attempt to increase the ovulation rate by increasing the interval between inhibin immunization and PGF2alpha injection. Five miniature Shiba goats were treated with 10 ml inhibin antiserum (inhibin-AS) developed against [Tyro30]-inhibin alpha (1-30). A control group (n=5) was treated with normal goat serum. All animals were injected intramuscularly with 125 microg PGF2alpha 72 h after treatment to induce estrus and ovulation. Blood samples were collected for hormonal assay and the ovulation rate was determined by laparotomy. In contrast to the control group, there was a significant increase in plasma concentrations of FSH in the immunized group. After luteolysis, plasma concentrations of estradiol-17beta increased markedly to a preovulatory peak about 2 folds higher (P<0.01) than that of controls. In addition, the ovulation rate was greater in the immunized group (14.4 +/- 2.2) than in the control group (2.2 +/- 0.6), and the mean number of follicles > or = 4 mm in diameter was 10.0 +/- 0.8 in the inhibin-AS group compared with 2.4 +/- 0.3 in control group. The present results demonstrate that immunoneutralization of endogenous inhibin increased FSH secretions in miniature shiba goats. The increased FSH secretion enhanced follicular growth and increased the ovulation rate. Additionally, increasing the interval between inhibin-AS and PGF2alpha injections (to 72 h) resulted in a greater ovulation rate compared with the previous protocol (48 h). Therefore, inhibin-AS treatment proved to be an effective alternative to exogenous gonadotropin methods for induction of superovulation in goats.

Animals↗

PubMed2000

Cerebello-olivary and lateral (accessory) cuneate degeneration in a juvenile American Miniature horse.

A 12-month-old American Miniature horse colt was presented to the Virginia Tech Veterinary Teaching Hospital with a 7-month history of progressive ataxia. Physical examination revealed a head intention tremor, base-wide stance, and ataxia. Necropsy findings were confined to the brain. There were bilateral areas of liquefactive necrosis and cavitation corresponding to the dorsal accessory olivary and lateral (accessory) cuneate nuclei. Cerebellar folia of the dorsal vermis were thin. Microscopically, the cerebellar cortex was characterized by patchy areas of Purkinje cell loss with associated variable thinning of the molecular and granule cell layers and astrogliosis. Dorsal accessory olivary and lateral cuneate nuclei were cavitated and had mild glial response around their periphery. Additionally, a focus of necrosis and neuropil vacuolization was found in the right putamen. These findings indicate the presence of a neurodegenerative disorder centered, but not confined to, the cerebellum and its connections in this American Miniature horse colt.

Animals↗

PubMed2004

Rapid-scanning forward-imaging miniature endoscope for real-time optical coherence tomography.

We developed a miniature endoscope that is capable of rapid lateral scanning and is suitable for real-time forward-imaging optical coherence tomography (OCT). The endoscope has an outer diameter of 2.4 mm, consisting of a miniature tubular lead zirconate titanate (PZT) actuator, a single-mode fiber-optic cantilever, and a graded-index lens. Rapid lateral scanning at 2.8 kHz is achieved when the fiber-optic cantilever is resonated with the PZT actuator. This allows OCT imaging to be performed by fast lateral beam scanning followed by slow depth scanning, which is different from the conventional OCT imaging sequence. Real-time OCT imaging with the endoscope operated in the new image acquisition sequence at 6 frames/s is demonstrated.

Computer Systems↗

PubMed2005

Mode transformer for miniaturized optical circuits.

A novel mode transformer was fabricated that transforms a modal area by a factor of 100. Using the mode transformer improves the efficiency of mode transformation by an order of magnitude compared with that when no mode transformer is used. With this mode transformer, input-output coupling of miniaturized, on-chip integrated optical circuits to external optical fibers is achieved with low loss. The mode transformer's design, fabricated in silicon, is scalable to virtually any waveguide size, facilitating continuous miniaturization in silicon optoelectronics.

Journal Article↗

PubMed2024

High-throughput DNA extraction and cost-effective miniaturized metagenome and amplicon library preparation of soil samples for DNA sequencing.

Reductions in sequencing costs have enabled widespread use of shotgun metagenomics and amplicon sequencing, which have drastically improved our understanding of the microbial world. However, large sequencing projects are now hampered by the cost of library preparation and low sample throughput, comparatively to the actual sequencing costs. Here, we benchmarked three high-throughput DNA extraction methods: ZymoBIOMICS™ 96 MagBead DNA Kit, MP BiomedicalsTM FastDNATM-96 Soil Microbe DNA Kit, and DNeasy® 96 PowerSoil® Pro QIAcube® HT Kit. The DNA extractions were evaluated based on length, quality, quantity, and the observed microbial community across five diverse soil types. DNA extraction of all soil types was successful for all kits, however DNeasy® 96 PowerSoil® Pro QIAcube® HT Kit excelled across all performance parameters. We further used the nanoliter dispensing system I.DOT One to miniaturize Illumina amplicon and metagenomic library preparation volumes by a factor of 5 and 10, respectively, with no significant impact on the observed microbial communities. With these protocols, DNA extraction, metagenomic, or amplicon library preparation for one 96-well plate are approx. 3, 5, and 6 hours, respectively. Furthermore, the miniaturization of amplicon and metagenome library preparation reduces the chemical and plastic costs from 5.0 to 3.6 and 59 to 7.3 USD pr. sample. This enhanced efficiency and cost-effectiveness will enable researchers to undertake studies with greater sample sizes and diversity, thereby providing a richer, more detailed view of microbial communities and their dynamics.

Metagenome↗

PubMed2004

Miniaturized multiplex label-free electronic chip for rapid nucleic acid analysis based on carbon nanotube nanoelectrode arrays.

BACKGROUND: Reducing cost and time is the major concern in clinical diagnostics, particularly in molecular diagnostics. Miniaturization technologies have been recognized as promising solutions to provide low-cost microchips for diagnostics. With the recent advancement in nanotechnologies, it is possible to further improve detection sensitivity and simplify sample preparation by incorporating nanoscale elements in diagnostics devices. A fusion of micro- and nanotechnologies with biology has great potential for the development of low-cost disposable chips for rapid molecular analysis that can be carried out with simple handheld devices. APPROACH: Vertically aligned multiwalled carbon nanotubes (MWNTs) are fabricated on predeposited microelectrode pads and encapsulated in SiO2 dielectrics with only the very end exposed at the surface to form an inlaid nanoelectrode array (NEA). The NEA is used to collect the electrochemical signal associated with the target molecules binding to the probe molecules, which are covalently attached to the end of the MWNTs. CONTENT: A 3 x 3 microelectrode array is presented to demonstrate the miniaturization and multiplexing capability. A randomly distributed MWNT NEA is fabricated on each microelectrode pad. Selective functionalization of the MWNT end with a specific oligonucleotide probe and passivation of the SiO2 surface with ethylene glycol moieties are discussed. Ru(bpy)2+ -mediator-amplified guanine oxidation is used to directly measure the electrochemical signal associated with target molecules. SUMMARY: The discussed MWNT NEAs have ultrahigh sensitivity in direct electrochemical detection of guanine bases in the nucleic acid target. Fewer than approximately 1000 target nucleic acid molecules can be measured with a single microelectrode pad of approximately 20 x 20 microm2, which approaches the detection limit of laser scanners in fluorescence-based DNA microarray techniques. MWNT NEAs can be easily integrated with microelectronic circuitry and microfluidics for development of a fully automated system for rapid molecular analysis with minimum cost.

Algorithms↗

PubMed1994

Usefulness of miniature flexible fiberoptic bronchoscopy in children.

A miniature flexible fiberoptic bronchoscope (FFB) (Olympus BF-N20) (2.2 mm diameter) was applied to 53 children (20 female subjects) ranging in age from 3 months to 15 years (mean, 4.19 years). Most common indications for bronchoscopy included stridor or weak cry and persistent wheezing or cough unresponsive to inhaled bronchodilators, chest physiotherapy, steroids, and antimicrobial agents. There were no complications. In 38 children (71.6 percent) it was diagnostically useful, particularly for the investigations of upper airway obstruction (66 percent). In 22 children (41.5 percent) it provided guidance for surgical interventions. The instrument was particularly useful during its application in infants with severe upper airway obstruction who otherwise would require open rigid-tube bronchoscopy in the operating room. It was of limited value when excessive bronchial secretions obstructed the view of the working field for which a bronchoscope with a built-in suction channel was needed. It is concluded that this miniature FFB is a useful diagnostic tool in infants and children particularly for obstructed upper airways but has limited applications in children with peripheral airway disease. The 2.2-mm bronchoscope may have its greatest advantage in preterm neonates and intubated infants, where the small glottic or endotracheal tube size renders the 3.5-mm bronchoscope useless.

Adolescent↗

PubMed1976

A fluidic-controlled, miniature respirator with a new positive airway pressure device.

A simple and miniature respirator was developed providing controlled ventilation and application of a positive airway pressure. The positive airway pressure is achieved by an airflow through a canula into the airways. During expiration the direction of this insufflated air-flow is changed and the dynamic pressure necessary for it manifests itself as a positive airway pressure. Mechanical ventilation functions according to the principle of the Ayre-T-Piece: flow direction and, therefore ventilation, is controlled by occlusion of the expiratory limb. Occlusion is brought about by inflating a balloon. The inflation of the balloon is controlled by fluidic-circuits. They allow setting of frequency, duration of inspiration and pressure limit. As the above mentioned system for creating a positive airway pressure is integrated, mechanical ventilation with PEEP is achieved. Tests on a lung model were performed to establish the variables which influence the amount of positive pressure produced by an air-flow through a canula into a tube. Results indicate that the positive pressure obtained is inversely correlated to the diameter of tube and canula and varies with the angle of inflow (Tab. I, II, Fig. 4). The pressure-rise-velocity depicted in Fig. 5 suggests that sighing is possible in an open system. On an anencephalic newborn pressure-flow-ratios were established (Fig. 6). Pressure-time curves during spontaneous breathing, CPAP assisted breathing and sighing combined with CPAP were recorded in a 3 month old baby (Fig. 7, 8, 9). The discussion states the technical performance of the respirator (frequency 100/min-1/3 min., tidal volume 5-100 ml, controlled ventilation with pressure limitation, application of PEEP) and additional advantages (no apparatus deadspace, disposable i.e. sterile air supply-system, the miniature dimensions 10 x 10 x 5 cm, versatility and simple therefore technically reliable construction).

Humans↗

PubMed2004

Durability and validity of a remote, miniaturized pressure sensor in an animal model of abdominal aortic aneurysm.

PURPOSE: To investigate whether a remote, miniaturized pressure sensor could maintain calibration and function through organized thrombus over an extended period in a porcine model of abdominal aortic aneurysm (AAA). METHODS: Six adult pigs had an AAA surgically created and excluded. A sensor zeroed to atmospheric pressure was placed within the aneurysm sac and another within the suprarenal aorta of each animal. Pressure measurements were taken at the initial operation and then on a weekly basis over 2 months. The aortic sensors were correlated to an intra-arterial pressure catheter at the initial operation and at the time of sacrifice. Back-table sensor correlation with atmospheric pressure was done at the time of explantation. RESULTS: Three animals died during the follow-up period. Five animals were available for 6-week follow-up, of which 3 survived for the complete 8-week protocol. Two of the surviving animals had an intra-aortic sensor. All 5 aneurysm sac sensors functioned throughout the experimental period. At the time of sacrifice, the sacs contained a large amount of organized thrombus in which the sac sensors were deeply embedded. The 3 aortic sensors also functioned throughout the course of the experiments. The pressures correlated within 5 mmHg to the catheter-based measurements taken at the initial operation and at the time of sacrifice. Comparison to atmospheric pressure revealed no calibration offset in any sensor. CONCLUSIONS: This chronic implantation study demonstrates the durability of a remote, miniaturized pressure sensor within a surgically created aneurysm sac as well as the suprarenal aorta of a porcine AAA model. There was no calibration offset in any of the sensors, and they remained valid at explantation. We believe that this is further evidence of the potential applicability of this sensor for clinical use.

Animals↗

PubMed2005

Glycoconjugate histochemistry of the secretory epithelium lining the seminal vesicles of the miniature pig.

The present study thoroughly examined the localization and characterization of glycoconjugates in the secretory epithelium lining the seminal vesicles of the miniature pig, employing light and electron microscopic histochemical procedures, including lectin methods. The present results showed the epithelial cells and luminal secretions to contain glycoconjugates with abundant neutral saccharides and a small amount of acidic saccharides, containing varying types of terminal sugar residues. At ultrastructural levels, the free surface coat of the plasma membrane was rich in alpha-D-Man, alpha-D-Glc, beta-D-Gal, GlcNAc, and sialic acid. The flocculent contents of the secretory vesicles indicated the localization of alpha-D-Man, alpha-D-Glc, alpha-L-Fuc, beta-D-Gal, GlcNAc, and sialic acid; such sugar residues were also seen in the elements of the Golgi apparatus. The present results have characterized the seminal vesicles of the miniature pig as having a high secretory activity and copiously producing glycoconjugates with various sugar residues. Such glycoconjugates appear to be indispensable substances for porcine reproduction, possibly influencing the fertilizing capacity of spermatozoa within the female genital tract.

Animals↗

PubMed2004

A miniaturized FIA system for the determination of residual chlorine in environmental water samples.

The present study was aimed at developing a portable miniaturized FIA system with a two-electrode based amperometric detector for on-site monitoring of residual chlorine in environmental samples. The electrode potential control and current data collection were conducted with a home-designed potentiostat with a size of 150 x 100 x 40 mm and the weight of 350 g. It was confirmed that the amperometric response increased in proportion to the chlorine concentration up to 5 microg/ml, and the detection limit for chlorine was 0.05 microg/ml. Besides a drastic reduction in the size of the analytical system, such miniaturization achieved the advantages of increased speed, minimal mobile-phase consumption, and high sensitivity. The proposed method was successfully applied to the determination of residual chlorine in samples of pool water and tap water. The results were well agreed with those obtained by the N,N-diethyl-p-phenylenediamine (DPD) photometric method.

Journal Article↗

PubMed2000

Fluorescence polarization is a useful technology for reagent reduction in assay miniaturization.

The use of fluorescence polarization (FP) has increased significantly in the development of sensitive and robust assays for high throughput screening of chemical compound libraries during the past few years. In this study, we show that FP is a useful assay miniaturization technology for reagent reduction during high throughput screening. We developed and optimized several FP assays for binding to estrogen receptor alpha and two protein kinases with an assay volume of 100 microl. Without any re-optimization, a consistent signal window was maintained in 384- or 1536-well format when the assay volume varied from 2.5-100 microl at constant concentrations of all assay components. In contrast, the signal window decreased with decreasing assay volume at constant reagent concentration in the protein kinase C scintillation proximity assay (SPA) and prompt fluorescence assay. In addition, the effect of evaporation on the signal window was minimal for the FP assays. Our study suggests that FP is superior to SPA and prompt fluorescence in terms of reagent reduction in the miniaturized assay format.

Biochemistry↗

PubMed2003

Lobar holoprosencephaly in a Miniature Schnauzer with hypodipsic hypernatremia.

A 9-month-old male Miniature Schnauzer was examined because of a lifelong history of behavioral abnormalities, including hypodipsia. Diagnostic evaluation revealed marked hypernatremia and a single forebrain ventricle. The behavioral abnormalities did not resolve with correction of the hypernatremia, and the dog was euthanatized. At necropsy, midline forebrain structures were absent or reduced in size, and normally paired forebrain structures were incompletely separated. Findings were diagnostic for holoprosencephaly, a potentially genetic disorder and the likely cause of the hypodipsia. Similar evaluation of affected Miniature Schnauzer dogs may reveal whether holoprosencephaly routinely underlies the thirst deficiency that may be seen in dogs of this breed.

Animals↗