An eye movement recording method operating with a closed eye

An eye movement recorder, based on an optoelectronic method, can be used in closed or open eye situations. Three photodiodes are used as proximity sensors. This eye tracker is well adapted for the studies of eye movement in after-image phenomena.     

Facilitated giga-seal formation with a just originated glass surface

A simple technique of tip preparation in patch pipettes is described, which facilitates giga-seal formation. The pipettes were fabricated from thick-walled borosilicate glass tubing (external diameter 2.0 mm, internal diameter 0.5 mm) and the tips could be repeatedly broken in the bath. The pipette resistance correspondingly fell in steps of 3–20 M from about 80 M to about 2 M (double concentrated Tyrode). Scanning electron microscopy showed that the tip obtained after breaking was fairly plain. These broken tips were especially appropriate for patch-clamping. In cardiac myocytes in 11 out of 26 patches with Na⁺ channel activity, giga-seals developed spontaneously, i.e. without suction. In these patches the amplitude of the mean current with depolarizing pulses to –40 mV was significantly higher in comparison with patches formed under negative pressure. It is concluded that spontaneously sealed patches are most likely of planar configuration and the Na⁺ channel activity exceeds that in suction-induced patches.     

A single unit microelectrode with a glass seal and a microhole

Summary A method is presented for the construction, calibration and utilization of a single unit ion-selective microelectrode of the internal capillary configuration. The microelectrode is ideally suited for in situ applications. The microelectrode capacity can be made as small as 10 nanoliters. It is superior to previously constructed microelectrode cells in that it combines the ion-selective and the reference electrodes into one physical unit.     

Performance evaluation of a robot-assisted catheter operating system with haptic feedback

In this paper, a novel robot-assisted catheter operating system (RCOS) has been proposed as a method to reduce physical stress and X-ray exposure time to physicians during endovascular procedures. The unique design of this system allows the physician to apply conventional bedside catheterization skills (advance, retreat and rotate) to an input catheter, which is placed at the master side to control another patient catheter placed at the slave side. For this purpose, a magnetorheological (MR) fluids-based master haptic interface has been developed to measure the axial and radial motions of an input catheter, as well as to provide the haptic feedback to the physician during the operation. In order to achieve a quick response of the haptic force in the master haptic interface, a hall sensor-based closed-loop control strategy is employed. In slave side, a catheter manipulator is presented to deliver the patient catheter, according to position commands received from the master haptic interface. The contact forces between the patient catheter and blood vessel system can be measured by designed force sensor unit of catheter manipulator. Four levels of haptic force are provided to make the operator aware of the resistance encountered by the patient catheter during the insertion procedure. The catheter manipulator was evaluated for precision positioning. The time lag from the sensed motion to replicated motion is tested. To verify the efficacy of the proposed haptic feedback method, the evaluation experiments in vitro are carried out. The results demonstrate that the proposed system has the ability to enable decreasing the contact forces between the catheter and vasculature.     

Communications for a Picture Archiving Communications System with a parallel operating image data base

In cooperation with BAZIS (Support Group Hospital Information Systems at Leiden, The Netherlands), we started the design of a Picture Archiving and Communications System (PACS) a few years ago that contains a centralized image data base. By means of system modeling and measuring some performance parameters, we have discovered that such a system cannot provide the required response times. This is mainly because of equipment communication interfaces that are not fast enough and have a throughput that does not generally exceed 16 Mbits/second. By providing a parallel operating image buffer system, the problems related to the limited speed of the communication interfaces can be overcome. Moreover, by splitting the network into a number of subnets connected to each other by means of “bridges,” the communications load can be locally reduced, improving in this manner the response times. Our PACS contains a supervisor and one relational data base that relates patient data with image locations. A significant part of the report is concerned with the design of the high speed American College of Radiology-National Electronic Manufacturing Association transmission control protocol/internet protocol (ACR-NEMA TCP/IP) communications interface. The use of such an interface eliminates the need for a Network Interface Unit, reducing in this manner communication delays and complexity. We have developed and have in operation for experimental purposes a small-scale PACS, consisting of a number of workstations, a relational data base and image buffers, all connected to a network that uses the ACR-NEMA TCP/IP protocols. This experimental PACS is used to provide input data to a PACS performance model, to validate this model, and to investigate alternative system configurations.     

Multifunctional magnetic mesoporous bioactive glass scaffolds with a hierarchical pore structure

Hyperthermia and local drug delivery have been proposed as potential therapeutic approaches for bone defects resulting from malignant bone tumors. The development of bioactive materials with magnetic and drug delivery properties may potentially meet this target. The aim of this study was to develop a multifunctional mesoporous bioactive glass (MBG) scaffold system for both hyperthermic and local drug delivery applications. To this end iron (Fe)-containing MBG (Fe-MBG) scaffolds with a hierarchical large pores structure (300–500 μm) and fingerprint-like mesopores (4.5 nm) have been prepared. The effects of Fe on the mesopore structure and physiochemical, magnetic, drug delivery and biological properties of MBG scaffolds have been systematically investigated. The results show that the morphology of the mesopores varied from straight channels to curved fingerprint-like channels after incorporation of Fe into MBG scaffolds. The magnetism of MBG scaffolds can be tailored by controlling the Fe content. Furthermore, the incorporation of Fe into mesoporous MBG glass scaffolds enhanced the mitochondrial activity and the expression of bone-related genes (ALP and OCN) in human bone marrow mesenchymal stem cells (BMSC) attached to the scaffolds. The Fe-MBG scaffolds obtained also possessed high specific surface areas and demonstrated sustained drug delivery. Thus Fe-MBG scaffolds are magnetic, degradable and bioactive. The multifunctionality of Fe-MBG scaffolds suggests that there is great potential for their use in the treatment and regeneration of large-bone defects caused by malignant bone tumors through a combination of hyperthermia, local drug delivery and osteoconductivity.     

Temporally regulated assembly of a dynamic signaling complex associated with the activated TCR

TCR triggering promotes multiple tyrosine kinase-dependent interactions involving proteins with one or more protein binding modules. Reported interactions mostly exceed the binding potential of these proteins. A solution to this paradox is the temporally regulated recruitment of alternative ligands. We have tested this hypothesis by analyzing the time course of protein/protein interactions triggered by TCR engagement. We show that a short-lived and dynamic multimolecular complex is assembled on tyrosine-phosphorylated CD3zeta. Specific components of this complex are recruited and shed in a temporal sequence distinct for each of the proteins analyzed. The temporally regulated assembly of a higher order structure at the activated TCR is likely to be crucial in achieving both signal longevity and signal specificity.     

Development of a miniaturepH glass electrode with field-effect-transistor amplifier for biomedical applications

A miniaturepH glass electrode for possible clinical and biomedical applications was constructed and evaluated in referencepH buffer solutions at 36°C. A field-effect-transistor (f.e.t.) amplifier was integrated into the system, and its effectiveness was investigated. The average e.m.f. efficiency of the developed electrode was 0·87±0·11V, and a minimum length of 7 mm was required for a goodpH response. Temperature dependence of the electrode was ?1·51 mV/^oK at apH of 7. Evaluation of the stability of the electrode showed a 1% drift over a 7 h operational time. ThepH and temperature hysteresis effects showed 0·5% and 1·0% deviations, respectively. The response time was within 4 s for 99% response. These electrode characteristics were also investigated in blood plasma and discussed.     

Outbreak of infection with a multiresistant Klebsiella pneumoniae strain associated with contaminated roll boards in operating rooms

An outbreak with a multiresistant Klebsiella pneumoniae (MRKP) strain among seven patients admitted to the adult intensive care unit (ICU) of a regional teaching hospital in The Netherlands was investigated. Epidemiologic investigations revealed a short delay between an operation and the acquisition of the MRKP strain. A case-control study comprising 7 cases and 14 controls was conducted to identify the risk factors associated with the acquisition of the MRKP strain. An operation at each of two operation rooms was strongly associated with the acquisition of the MRKP strain: odds ratio of 36 (95% confidence interval, 2.7 to 481.2; P=0.003, Fisher exact two-tailed test). Cultures of environmental specimens of the operation rooms revealed contamination of the roll boards used to transport patients from the bed to the operation table with the MRKP strains. Molecular genotyping of the isolates revealed clonal similarity between the isolates of the seven cases, isolates from environmental specimen cultures, and in addition, an MRKP isolate from a re-patriated ICU patient from earlier that year. The outbreak ended after cleaning and replacement of the roll boards in the operation rooms and implementation of additional barrier precautions for colonized or infected patients. It was concluded that two operation rooms played a significant role in the transmission of an MRKP strain between ICU patients during the presented outbreak.     

The interface of various glasses and glass ceramics with a bony implantation bed

Basing on histomorphological evaluations and morphometrical quantifications in a standardized model experiment, a comparison is made between the reactions of skeletal tissues to various glasses, glass-ceramics and enamels. On the surface of these so-called reactive biomaterials either a direct bonding to mineralized bone or also different amounts of osteoid or chondroid tissue formation can be observed, depending on the composition of the material. It is shown that the solubility of the glasses cannot directly be related to the reactivity and the resulting bone bonding; bone binds only to glasses with a controlled release of constituents and which exhibit a seam of extracellular matrix on their surface, in which normal primary mineralization can occur; the release of constituents such as Al2O3, Ta2O5, ZrO2, or phosphates from the material can inhibit this normal mineralization and the transformation of chondroid tissue to bone; if connective tissue instead of bone is present at the interface (either primarily or after bone remodelling), the dissolution (or corrosion) of the material may be no longer controllable, and the tissue reacts with a continuous inflammatory response to the corrosion products.     

Novel genetic instability associated with a developmental regulated glycosyltransferase locus in Chinese hamster ovary cells

LEC10 is a dominant glycosylation mutant of Chinese hamster ovary (CHO) cells that expresses a developmentally regulated glycosyltransferase (GlcNAc-TIII) not detectable in parental CHO cells. Several mutagens were found to increase the frequency of LEC10 mutants up to 10-fold over the spontaneous frequency of 10^–7, while 5azaC treatment had no effect. Revenants were obtained at high frequency (10^–4) and were found to belong to two classes. Three independent revertants gave rise to new LEC10 mutants at high frequency (10^–4) while seven others gave new LEC10 mutants at the low frequency typical of unmutagenized parental CHO cells. No evidence of a general mutator phenotype was found in the revenant lines with a high rereversion frequency. The combined data suggest a novel form of genomic instability at the LEC10 locus in CHO cells. Genetic events that affect the expression of developmentally regulated glycosyltransferases may be identified by further studies of LEC10 and other dominant CHO glycosylation mutants.     

A developmentally regulated heparan sulfate epitope defines a subpopulation with increased blood potential during mesodermal differentiation

Heparan sulfate (HS) is a mandatory coreceptor for many growth factors and morphogens involved in embryonic development; its bioactivity is dictated by complex sulfation motifs embedded within the polymer chain. Using a panel of HS-specific antibodies we have identified a unique HS epitope recognized by antibody HS4C3 that is selectively expressed during differentiation of embryonic stem (ES) cells along the mesodermal lineage to the hemangioblast stage. The appearance of this high-affinity HS4C3-binding (HS4C3(high)) epitope is transient; the epitope is specifically expressed within the emerging Brachyury(+) (Bry(+)) population and marks those cells that will become fetal liver kinase 1 (Flk1)(+). Fluorescence-activated cell sorting (FACS) separation and colony forming assays revealed that HS4C3(high)/Flk1(+) cells have a dramatically increased potential to form both blast and endothelial colonies, both of which depend upon the HS-binding growth factor vascular endothelial growth factor. Critically, expression of this HS epitope is tightly regulated, disappearing from the cell surface as the resultant hematopoietic lineages mature, in a similar manner to protein markers Bry and Flk1. In vivo studies showed a remarkable correlation with in vitro findings, with expression of HS4C3-binding epitopes restricted to newly formed mesodermal tissues during gastrulation. We believe this is the first time a defined HS epitope has been implicated in a specific developmental pathway and that this provides, in addition, a novel enrichment technique for the isolation of hemangioblasts from mixed differentiated ES cell cultures.     

Bone bonding in bioactive glass ceramics combined with a new synthesized agent TAK-778

We studied the stimulatory effects of TAK-778, a new synthetic 3-benzothiepin derivative that promotes osteoblast differentiation, in the bonding of bone to bioactive glass ceramic implants in rabbit tibiae. Smooth-surfaced, rectangular plates (15 x 10 x 2 mm) made of apatite-wollastonite-containing glass ceramic were implanted bilaterally into the proximal metaphyses of rabbit tibiae. Sustained-release microcapsules containing TAK-778 were packed into the medullary cavity in one limb and untreated microcapsules were packed into the contralateral limb to serve as a paired control. At 4, 8, and 16 weeks after implantation, bonding at the bone/implant interfaces was evaluated using a detaching test and histological examination of undecalcified specimens. The tensile failure load increased during weeks 4 to 16 in both groups; the tensile failure load in the TAK-778-treated group was significantly greater than that in the control group at each interval after implantation. Histologically, the TAK-778-treated specimens showed greater active new bone formation mainly in the medullary cavity and more extensive bonding between the implant and bone than the untreated specimens. The results of this study suggest that adding the bone formation-promoting TAK-778 to bioactive glass ceramic implants may significantly accelerate bone apposition to the implants and improve the bonding process at the interface. This would help to establish earlier and stronger bonding of orthopedic ceramic implants to the surrounding bone tissue.     

Bioactive glass as a drug delivery system of tetracycline and tetracycline associated with beta-cyclodextrin

The aim of this study was to evaluate the physical-chemical properties, in vivo biocompatibility and antimicrobial activity of bioactive glasses (BG) used as a controlled release device for tetracycline hydrochloride and an inclusion complex formed by tetracycline and beta-cyclodextrin at 1:1 molar ratio. The BG as well as their compounds loaded with tetracycline (BT) and tetracycline:beta-cyclodextrin (BTC) were characterized by FTIR spectroscopy, X-ray powder diffraction, differential scanning calorimetry and by scanning electron microscopy and energy dispersive spectroscopy. The in vivo test was carried out with female mice split into three groups treated with bioactive glass either without drugs, or associated with tetracycline, or with tetracycline:beta-cyclodextrin by subcutaneous implantation. The histological examination of tissue at the site of implantation showed moderate inflammatory reactions in all groups after 72 h. The bacterial effect was tested on A. actinomycetemcomitans suspended in BHI broth, with or without bioactive particles. A considerable bacteriostatic activity was found with BT and BTC glasses, as compared to plain glass. The presence of cyclodextrin was important to slow down the release of tetracycline for a long period of time and it was verified that the presence of tetracycline or its inclusion complex, tetracycline:beta-cyclodextrin, did not affect the bioactivity of the glass.