Anesthesia for laparoscopic surgery in a patient with myotonic dystrophy (Steinert's disease): beneficial use of sugammadex, but incorrect use of pethidine: a case report.

Patients with Myotonic Dystrophy show an unpredictable response to several anesthetic drugs including opioids, neuromuscular blocking agents and especially reversal agents like neostigmine. We describe the case of a 40 year old patient with myotonic dystrophy who underwent laparoscopic cholecystectomy and ovarian cyst removal under general anesthesia. The authors suggest the use of the new reversal agent suggamadex, for reversing neuromuscular blockade caused by rocuronium, in patients suffering from neuromuscular disease and especially from Myotonic Dystrophy, because it rapidly and completely reverses any residual neuromuscular blockade, but also underline the increased susceptibility of these patients to opioids.

Erythropoietin encapsulation in chitosan nanoparticles and kinetics of drug release.

Recombinant human erythropoietin (rHu-EPO) is a glycoprotein, which is produced commercially from Chinese hamster ovary (CHO) cells. It is used for the therapy of renal anemia and chemotherapy-induced anemia in cancer patients. Recent evidence suggests that rHu-EPO exerts tissue protective effects via multiple mechanisms which include inhibition of apoptosis, promotion of angiogenesis and decreased inflammation. After intravenous (i.v.) injection, the blood concentration of rHu-EPO rapidly decreases due to proteolysis resulting in a relatively short half-life of 8.5 h, which necessitates regular dosing with intervals that do not exceed 7 days. It would be desirable to develop an encapsulated formulation providing controlled release of rHu-EPO to maintain therapeutic concentrations in plasma, and for potential tissue protective applications to maintain high local therapeutic concentrations in tissue while minimizing potential unwanted systemic effects such as polycythemia and platelet activation, both of which can predispose to intravascular thrombosis. Nanoparticle encapsulation of rHu-EPO can also allow for direct injection at sites of injury in specific tissues/organs, again minimizing systemic exposure of the drug. In this paper, we report the production of biopolymer nanoparticles by ionotropic gelation of chitosan with tripolyphosphate (TPP). The nanoparticle size distribution in aqueous solution was determined and rates of rHu-EPO release from chitosan-TPP nanoparticles were measured in PBS at 37°C. It was observed that almost 30% of the encapsulated rHu-EPO was released within the first 48 hours and thereafter a linear release profile was observed for up to 2 weeks. Total drug release over 15 days was 63% of the initial amount.

A case of recurrent benign lymphocytic (Mollaret's) meningitis and review of the literature.

Mollaret's meningitis is a rare form of benign recurrent aseptic meningitis first described in 1944. We report a case of Mollaret's meningitis due to Herpes Simplex Virus type 2 (HSV2), diagnosed with Polymerase Chain Reaction (PCR) implementation in the Cerebrospinal fluid (CSF) of the patient and treated successfully with acyclovir. To our knowledge, this is the first case of Mollaret's meningitis reported in Greece. We reviewed the literature since PCR has become widely available. Herpes Simplex Virus type 2 has been the most commonly identified causative agent of Mollaret's meningitis.

Biodegradation of petroleum hydrocarbons in an immobilized cell airlift bioreactor.

An "immobilized cell airlift bioreactor", was used for the aerobic bioremediation of simulated diesel fuel contaminated groundwater and tested with p-xylene and naphthalene in batch and continuous regimes. The innovative design of the experiments consists of two stages. At the first stage "immobilized soil bioreactor" (ISBR) was used to develop an efficient microbial consortium from the indigenous microorganisms, which exist in diesel fuel contaminated soil. The concept of ISBR relies on the entrapment of the soil particles into the pores of a semi-permeable membrane, which divides the bioreactor into two aerated and non-aerated portions. The second stage involves inoculating the "immobilized cell air lift bioreactor" with the cultivated microbial consortia of the first stage. Immobilized cell airlift bioreactor has the same configuration as ISBR except that in this bioreactor instead of soil, microorganisms were immobilized on the fibers of the membrane. The performance of a 0.83 L immobilized cell airlift bioreactor was investigated at various retention time (0.5-6 h) and concentrations of p-xylene (15, 40 and 77 mg/L) and naphthalene (8, 15 and 22 mg/L) in the continuous operation. In the batch regime, 0.9L bioreactor was operated at various biodegradation times (15-135 min) and concentrations of p-xylene (13.6, 44.9 and 67.5 mg/L) and naphthalene (1.5 and 3.8 mg/L). Under the conditions of the complete biodegradation of p-xylene and naphthalene, the obtained volumetric biodegradation rates at biomass density of 720 mg/L were 15 and 16 mg/L h, respectively.

Nucleic acid technology screening of Australian blood donors for hepatitis C and human immunodeficiency virus-1 RNA: comparison of two high-throughput testing strategies.

BACKGROUND AND OBJECTIVES: The aim of this study was to compare the performance of two high-throughput strategies for hepatitis C virus (HCV) and human immunodeficiency virus-1 (HIV-1) RNA nucleic acid technology (NAT) screening in a volunteer blood-donor population. MATERIALS AND METHODS: The semiautomated Chiron Procleix HIV-1/HCV transcription mediated amplification (TMA) assay was used to screen 1 439 765 donations in two different testing configurations. Three sites (termed PDT sites) performed a mixture of individual donation (ID) and minipool (MP) testing, where 1 113 288 donations were screened as pools of 24 and an additional 32 003 donations were screened in ID format. A further two sites (termed SDT sites) screened 294 474 donations exclusively in ID format. RESULTS: A significantly higher proportion of initial NAT reactives that failed to react on follow-up testing [termed non-repeatably reactive (NRR)] was observed for ID testing at SDT sites than at PDT sites (0.082 vs. 0.047%: P < 0.01). Within the PDT sites, however, there was no significant difference between the NRR rate for MP or ID samples (0.037 vs. 0.047%; not significant). There was a significant difference in failed run rates between PDT and SDT sites (P < 0.01), with PDT sites having a higher run failure rate owing to non-amplification of the internal control. The PDT sites also had a significantly higher overall invalid sample rate. However, the invalid sample rate, specifically caused by known equipment failure, was significantly higher in the SDT sites, possibly attributable to greater usage (P < 0.0001). Four HCV NAT-positive/antibody-negative samples were identified in the course of the study. CONCLUSIONS: The comparison of the performance of PDT with SDT sites identified only minor differences that did not adversely impact on the timely release of blood products. Although both ID and MP strategies showed excellent specificity, irrespective of site configuration, the significantly increased NRR rate, observed exclusively for ID testing performed at SDT sites, indicates a potential for contamination that may limit the number of samples that can optimally be processed using ID testing. The performance data for ID testing in particular should serve as a useful benchmark for evaluating candidate NAT systems that are fully automated.

Effect of particle-particle shearing on the bioleaching of sulfide minerals.

The biological leaching of sulfide minerals, used for the production of gold, copper, zinc, cobalt, and other metals, is very often carried out in slurry bioreactors, where the shearing between sulfide particles is intensive. In order to be able to improve the efficiency of the bioleaching, it is of significant importance to know the effect of particle shearing on the rate of leaching. The recently proposed concept of ore immobilization allowed us to study the effect of particle shearing on the rate of sulfide (pyrite) leaching by Thiobacillus ferrooxidans. Using this concept, we designed two very similar bioreactors, the main difference between which was the presence and absence of particle-particle shearing. It was shown that when the oxygen mass transfer was not the rate-limiting step, the rate of bioleaching in the frictionless bioreactor was 2.5 times higher than that in a bioreactor with particle friction (shearing). The concentration of free suspended cells in the frictionless bioreactor was by orders of magnitude lower than that in the frictional bioreactor, which showed that particle friction strongly reduces the microbial attachment to sulfide surface, which, in turn, reduces the rate of bioleaching. Surprisingly, it was found that formation of a layer of insoluble iron salts on the surface of sulfide particles is much slower under shearless conditions than in the presence of particle-particle shearing. This was explained by the effect of particle friction on liquid-solid mass transfer rate. The results of this study show that reduction of the particle friction during bioleaching of sulfide minerals can bring important advantages not only by increasing significantly the bioleaching rate, but also by increasing the rate of gas-liquid oxygen mass transfer, reducing the formation of iron precipitates and reducing the energy consumption. One of the efficient methods for reduction of particle friction is ore immobilization in a porous matrix.

Poly(glutamic acid) for biomedical applications.

Paclitaxel is a widely used anti-cancer agent. Conjugates of paclitaxel with poly(glutamic acid) have shown great promise in preclinical trials, and clinical trials are now underway. Preclinical data suggest that more paclitaxel is preferentially delivered to tumor sites vs. nonconjugated paclitaxel. When poly(glutamic acid) is conjugated to other families of cancer drugs, similar improvements in effectiveness and reduced toxicity are observed. Optimization of poly(glutamic acid) for use in drug delivery applications is a key step in making this technology viable.

Hydrodynamic and mass transfer characteristics of three-phase gaslift bioreactor systems.

This review focuses on the hydrodynamic and mass transfer characteristics of various three-phase, gaslift fluidized bioreactors. The factors affecting the mixing and volumetric mass transfer coefficient (k(L)a), such as liquid properties, solid particle properties, liquid circulation velocity, superficial gas velocity, bioreactor geometry, are reviewed and discussed. Measurement methods, modeling and empirical correlations are reviewed and compared. To the authors' knowledge, there is no 'generalized' correlation to calculate the volumetric mass transfer coefficient, instead, only 'type-specific' correlations are available in the literature. This is due to the difficulty in modeling the gaslift bioreactor, caused by the variation in geometry, fluid dynamics, and phase interactions. The most important design parameters reported in the literature are: gas hold-up, liquid circulation velocity, 'true' superficial gas velocity, mixing, shear rate, aeration rate and volumetric mass transfer coefficient, k(L)a.

Expression of single chain antibodies (ScFvs) for c-myc oncoprotein in recombinant Escherichia coli membranes by using the ice-nucleation protein of Pseudomonas syringae.

The ice nucleation protein (INP) is a glycosyl phosphatidylinositol anchored outer membrane protein found in certain Gram-negative bacteria. In this study, the INP from Pseudomonas syringae was applied as a fusion partner with the single chain antibody fragment (ScFv) against the human oncoprotein c-myc. Two new plasmids pNinaZ-myc and pNinaZScFv-BsaA1 were constructed and cloned into Escherichia coli JM109. The expression of the fusion protein was successfully demonstrated in the cloned cells. The fusion proteins had no effect on the viability of the host cells. Ice nucleation activity measurements and flow cytometry studies were followed to investigate the membrane expression of the fusion protein.

A new kinetic model of protein adsorption on suspended anion-exchange resin particles.

The kinetics of adsorption of bovine serum albumin on an anion-exchange resin were measured in a batch system using a flow cell and ultraviolet absorbance, as a function of initial liquid-phase protein concentration and solid-to-liquid phase ratio. A new mathematical model for adsorption kinetics is presented that fits the experimental data to give a highly linear relationship with time, following a short transient period. Numerical integration of the differential form of the new composite nonlinear (CNL) kinetic model, containing three independent parameters, is shown to describe the dynamics of batch adsorption much better than alternative lumped parameter models. Although the new model is phenomenological rather than mechanistic, its principal parameter is shown to be a direct linear function of a physically measurable quantity. This study demonstrates that the model can accurately simulate protein concentration-time profiles using parameter estimates derived from correlations over a wide range of initial protein concentrations and phase ratios. The new CNL model is shown to be considerably superior to the Langmuir and solid-film linear kinetic models in this regard, having the additional advantage that an equilibrium isotherm for the system is not required.

Measurement of Gd-DTPA diffusion through PVA hydrogel using a novel magnetic resonance imaging method.

Polyvinyl alcohol-cryogel (PVA-C) is a hydrogel that is an excellent tissue mimic. In order to characterize mass transfer in this material, as well as to demonstrate in principle the ability to noninvasively measure solute diffusion in tissue, we measured the diffusion coefficient of the magnetic resonance (MR) contrast agent gadolinium diethylene triaminopentaacetic acid (Gd-DTPA) through PVA-C using a clinical MR imager. The method involved filling thick-walled rectangular PVA-C "cups" with known concentrations of Gd-DTPA solutions. Then by using a fast inversion recovery spin echo MR imaging protocol, a signal "null" contour was created in the MR image that corresponded to a second, known concentration of Gd-DTPA. By collecting a series of MR images through the PVA-C wall as a function of time, the displacement of this second known isoconcentration contour could be tracked. Application of Fick's second law of diffusion yielded the diffusion coefficient. Seven separate experiments were performed using various combinations of initial concentrations of Gd-DTPA within the PVA-C cups (3.2, 25.6, or 125 mM) and tracked isoconcentrations contours (0.096, 0.182, or 0.435 mM Gd-DTPA). The experimental results and the predictions of Fick's law were in excellent agreement. The diffusivity of Gd-DTPA through 10% PVA hydrogel was found to be (2.6 +/- 0.04) x 10(-10) m(2)/s (mean +/- s.e.m.). Separate permeability studies showed that the diffusion coefficient of Gd-DTPA through this hydrogel did not change with an applied pressure of up to 7.1 kPa. Accurate measurements could be made within 30 min if suitable Gd-DTPA concentrations were selected. Due to the excellent repeatability and fast data acquisition time, this technique is very promising for future in vivo studies of species transport in tissue.

Bubble rise velocities and drag coefficients in non-Newtonian polysaccharide solutions.

Microbially produced polysaccharides have properties which are extremely useful in different applications. Polysaccharide producing fermentations start with liquid broths having Newtonian rheology and end as highly viscous non-Newtonian solutions. Since aerobic microorganisms are used to produce these polysaccharides, it is of great importance to know the mass transfer rate of oxygen from a rising air bubble to the liquid phase, where the microorganisms need the oxygen to grow. One of the most important parameters determining the oxygen transfer rate is the terminal rise velocity of air bubble. The dynamics of the rise of air bubbles in the aqueous solutions of different, mostly microbially produced polysaccharides was studied in this work. Solutions with a wide variety of polysaccharide concentrations and rheological properties were studied. The bubble sizes varied between 0.01 mm3 and 10 cm3. The terminal rise velocities as a function of air bubble volume were studied for 21 different polysaccharide solutions with different rheological properties. It was found that the terminal velocities reached a plateau at higher bubble volumes, and the value of the plateau was nearly constant, between 23 and 27 cm/s, for all solutions studied. The data were analyzed to produce the functional relationship between the drag coefficient and Reynolds number (drag curves). It was found out that all the experimental data obtained from 21 polysaccharide solutions (431 experimental points), can be represented by a new single drag curve. At low values of Reynolds numbers, below 1.0, this curve could be described by the modofoed Hadamard-Rybczynski model, while at Re > 60 the drag coefficient was a constant, equal to 0.95. The latter finding is similar to that observed for bubble rise in Newtonian liquids which was explained on the basis of the "solid bubble" approach.

Determination of the volumetric mass transfer coefficient (k(L)a) using the dynamic "gas out-gas in" method: Analysis of errors caused by dissolved oxygen probes.

There are many dynamic methods for measuring the volumetric mass transfer coefficient. The "gas out-gas in" method can directly determine the volumetric mass transfer coefficient in a bioreactor system and provide estimates of the volumetric microbial oxygen uptake rate and the average oxygen saturation concentration at the gas-liquid interface. The errors on these parameters are large if the dissolved oxygen probe response time is not considered. For reliable measurements, deconvolution of the oxygen probe measurements must be made. (c) 1995 John Wiley & Sons, Inc.

Kinetics of sterilization of Lactobacillus brevis cells by the application of high voltage pulses.

The technique of irreversible electroporation has been successfully applied to cause a lethal effect on Lactobacillus brevis cells suspended in phosphate buffer solution, Na(2)HPO(4)/NaH(2)PO(4) . H(2)O (0.845/0.186 mM) between parallel plane electrodes. Tests were carried out at different temperatures (24,45,60, and 80 degrees C) to determine if there was a synergistic effect of temperature and electric pulse treatment on the destruction of L. brevis. Experimental results indicate that the viability (log N/N(0); where N(0) and N are the number of cells survived per milliliter before and after pulse voltage application, respectively) of L. brevis decreased with electric field strength E and temperature T and treatment time t(t). The relations between log(N/N(0)) and t(t) and log(N/N(0)) and E indicate that higher field strengths are more effective than higher treatment times in causing destruction of L. brevis cells. It was also found that as the temperature of the liquid medium containing L. brevis cells increased from 24 to 60 degrees C, the death rate of L. brevis cells increased with a decrease in the total treatment time t(t) (pulse width x number of pulses applied). The application of an electric field strength E = 25 kV/cm at 60 degrees C and treatment time t(t) = 10 ms resulted in very high destruction levels of L. brevis cells (N/N(0) = 10(-9)). In comparison with existing steam sterilization technology, this new method of sterilization using relatively low temperature and short treatment time could prove to be an excellent method to minimize thermal denaturation of important nutrient components in liquid media.

Preparation of novel cyclosporin A derivatives.

The hydroxyl group on the 2-N-methyl-(R)-((E)-2-butenyl)-4-methyl-L-threonine residue of cyclosporin A was protected by acetylation, then the double bond on the same amino acid residue was oxidatively cleaved using a periodate/permanganate reagent. The resultant derivative of cyclosporin A contained a carboxylic acid group which was subsequently reacted with the nucleophiles 5-(aminoacetamido)fluorescein and poly(L-lysine), in the presence of 1-ethyl-3-[3-(dimethylamino)propyl] carbodiimide, to furnish novel cyclosporin A conjugates.

Principles and biotechnological applications of bacterial ice nucleation.

Certain aerobic, Gram-negative bacteria, including the epiphytic plant pathogen, Pseudomonas syringae, possess a membrane protein that enables them to nucleate crystallization in supercooled water. Currently, these ice-nucleating (IN) bacteria are being used in snow making and have potential applications in the production and texturing of frozen foods, and as a replacement of silver iodide in cloud seeding. A negative aspect of these IN bacteria is frost damage to plant surfaces. Thus, of the various types of biological ice nucleators, bacteria have been the subject of most research and also appear relevant to the anticipated practical uses. The intent of this review is to explain the identification and ecology of the ice-nucleating bacteria, as well as to discuss aspects of molecular biology related to ice nucleation and consider existing and potential applications of this unique phenomenon.

A novel technique for measuring solute diffusivities in entrapment matrices used in immobilization.

A novel technique has been developed for measuring effective solute diffusivities in entrapment matrices used for cell immobilization. In this technique radiotracers were used to measure effective diffusivities and equilibrium partition coefficients of the solute between the liquid and solid matrix. Ca-alginate was used in this study, because it is one of the most commonly employed matrices for the immobilization of microbial, plant and mammalian cells. The experimental apparatus consisted of a single spherical Ca-alginate bead which was attached to a rotating rod and immersed in water containing C(14)-glucose. The rotational speed of the spherical bead was controlled and resulted in excellent mixing, and negligible external film mass transfer resistance, which allowed the measurement of true effective solute diffusivity within the solid matrix. The rates of C(14)-glucose diffusion within the Ca-alginate sphere were measured using a scintillation spectrometer. A mathematical model of unsteady-state diffusion in a sphere was used with appropriate boundary conditions, and the effective diffusivity of glucose was found from the best fit of the experimental data using a computer regression analysis method. Using 2% (w/v) Ca-alginate beads in this new radiotracer technique the effective diffusivity and partition coefficient of glucose were found to be 6.62 x 10(-10) m(2)/s and 0.98, respectively. The accuracy, advantages, and simplicity of this new method for diffusivity measurements are also compared to other existing methods.