Biomedical subjects

A Prokop

Publications and source records attributed to A Prokop.

At least 55 records · Page 3Linked to original sources

PubMed2000

[Magnetic resonance tomography in the diagnosis of intra-articular tibial plateau fractures: value of fracture classification and spectrum of fracture associated soft tissue injuries].

PURPOSE: To compare magnetic resonance imaging (MRI) and X-ray tomography in assessing the type of fracture, degree of comminution and amount of articular surface depression in acute tibial condylar fractures and to describe the associated soft tissue injuries diagnosed with MRI. METHOD: 27 patients with acute tibial plateau fractures were investigated using linear X-ray tomography and MRI employing T1-weighted and proton density turbo spin echo, STIR, and T2-weighted gradient echo images. Fractures were classified according to the AO classification system. The degrees of depression and comminution were measured and soft tissue injuries were recorded. RESULTS: Fractures were classified as type B1 in 7, as B2 in 6, and as B3 in 6 cases by MRI. More complex C-type fractures were diagnosed in 8 cases. MR and X-ray grading were consistent with the exception of two B3 fractures, which were graded as B1 by X-ray tomography. X-ray tomography under-estimated the degree of comminution 63% of the patients had either meniscal tears or complete ruptures of their cruciate or collateral ligaments. Ten meniscal tears were diagnosed in 9 of 27 patients. Complete tears of the anterior cruciate ligament were seen in 4, and avulsions of the posterior cruciate ligament in 2 patients. CONCLUSION: MRI allows a detailed assessment of acute tibial plateau fractures and can replace conventional X-ray tomography. The high rate of fracture-associated soft tissue lesions makes MRI an especially valuable tool.

Adult↗

PubMed2000

Sonic wave separation of invertase from a dilute solution to generated droplets.

It has previously been shown that a droplet fractionation process, simulated by shaking a separatory funnel containing a dilute protein solution, can generate droplets richer in protein than present in the original dilute solution. In this article, we describe an alternative method that can increase the amount of protein transferred to the droplets. The new method uses ultrasonic waves, enhanced by a bubble gas stream to create the droplets. The amount of protein in these droplets increases by about 50%. In this method, the top layer of the dilute protein solution (of the solution-air interface) becomes enriched in protein when air is bubbled into the solution. This concentrating procedure is called bubble fractionation. Once the protein has passed through the initial buildup, this enriched protein layer is transferred into droplets with the aid of a vacuum above the solution at the same time that ultrasonic waves are introduced. The droplets are then carried over to a condenser and coalesced. We found that this new method provides an easier way to remove the protein-enriched top layer of the dilute solution and generates more droplets within a shorter period than the separatory funnel droplet generation method. The added air creates the bubbles and carries the droplets, and the vacuum helps remove the effluent airstream from the condenser. The maximum partition coefficient, the ratio of the protein concentration in the droplets to that in the residual solution (approx 8.5), occurred at pH 5.0.

Glycoside Hydrolases↗

PubMed2000

Effect of shear on human insulin in zinc suspension.

Human insulin in zinc suspension was used as a model protein to test the effect of shear on the settling rate of proteins, a possible inference for protein denaturation. The rate of settling was determined directly in a spectrophotometer. Shear effects are important in retaining the activity of proteins and are present in bubble, foam, and droplet protein fractionation processes. A simple test, such as that conducted here, may even be useful for monitoring changes in protein structure caused by commercial shipping of the protein. The settling rate for insulin was continuously monitored in the original bottle by spectrophotometric absorbance changes as a function of time. A settling curve was determined following each shear experiment, which included shaking the "worked" insulin solution in a vortex mixer for different lengths of time. It was determined, when comparing long shaking times with short ones, that the initial settling rate was less for the long-term shaking of the insulin samples and greater for the short-term shaking. The secondary effects of light and heat, along with shaking, apparently did not produce differences from shaking alone.

Drug Stability↗

PubMed2000

Effect of protein denaturation on void fraction in foam separation column.

Foam fractionation is a cost-effective process that uses air to extract protein from a liquid (in this case "crude" dilute egg-albumin solution). This article deals with how the void fraction (fraction of air in the aerated solution) of foam is affected by heat denaturation of the protein. A 2-mm glass tube was used to sample the foam-liquid interface fluid in a 35-mm-diameter column in order to detect small changes in void fraction and foam production, which are not easily detected directly from the bulk foam. The main control variable in this study was the protein solution preheating time. As the preheating time increased, the initial void fraction in the column decreased. The initial void fraction of the undenatured solution ranged from about 0.73 to 0.80, and the void fraction for significant preheating times of 5 min ranged from approx 0.68 to 0.72. Furthermore, the period of foam production increased from 5 to 7 min for undenatured proteins in solution to as long 15 min for 5-min preheated solutions. Side-port sampling through a small capillary tube has the potential to be used as a rapid and inexpensive way to determine the level of protein denaturation by directly determining the void fraction and then estimating the effect of denaturation from a protein denaturation calibration curve of the void fraction.

Biotechnology↗

PubMed2000

[Results of kidney transplantation in Krakow in 1992-2000].

The aim of the study was an analysis of renal transplantation results in the Krakow Transplant Center during 1992-2000. The analysis concerned 94 cadaveric transplant recipients. The study group included 31 females aged 23 to 61 years (mean 40.4 years) and 63 males aged 16 to 60 years (mean 41.8 years). The time of pre-transplant renal replacement therapy ranged from 4 to 120 months (mean 32 months). The mean time of total ischaemia was 22 hours 20 minutes. The majority of the recipients had three identical antigens out of six typed. Most of the recipients were treated with three immunosuppressive drugs including: Cyclosporine A, Azathioprine and steroids. Immediately after kidney transplantation 25.6% of the patients had urine output and did not require dialysis. Acute renal failure (ARF) of the graft was observed in 73.2% recipients. The average number of hemodialysis sessions in patients presenting ARF was 10. Acute rejection was diagnosed in 41.5% of the patients. The most frequent complications were: CMV (cytomegalovirus) infection, UTI (urinary tract infection) and policytemia. In the study group 1-year survival rate of the patients was 97.8% and 1-year graft survival was 93.61%. The 5-year survival rates both in the patients and the grafts were very satisfactory (96.96% and 87.7% respectively).

Acute Kidney Injury↗

PubMed1999

[Value of MRI in diagnosis of occult fractures].

The term "occult fracture" defines the lack of visible fracture signs in conventional radiography although a fracture exists. The diagnosis is made in the sequel or through further diagnostic procedures. The use of scintigraphy and CT examinations is limited by their moderate sensitivity and specificity. In contrast, MRI is a diagnostic tool that allows early diagnosis of a fracture and of accompanying chondral or ligament damage. We report on 23 patients with traumatic fractures at 5 different anatomical sites, all of which were negative by x-ray, but the diagnosis could subsequently be made with MRI.

Adolescent↗

PubMed1999

[Are there guidelines for treatment of metacarpal fractures? Personal results and literature analysis of the last 12 years].

As the treatment of metacarpal fractures is today still a controversial subject, we conducted an analysis of the literature in order to present the different therapy guidelines, indications, and their results. The data from the follow-up of 1602 patients was taken from literature which dated from 1 January 1984 to 31 March 1996. A total of 522 patients who underwent surgery received K-wires, screws or external minifixateur. The conservative approaches ranged from immobilization to various methods of mobilization with different aids or without fixation of the fracture. The mobilization in a brace provided good to excellent results in 95% of the cases; however, the failure rate of therapy was 23% because of local bruises and skin necrosis (3%). The mobilization with handcast, tape etc. attained good to excellent results in 94% of the cases. Here, no complications occurred. Both the immobilization treatment and the surgery provided good to excellent results in 85% of the cases. The reposition of fractures of the fifth metacarpal was successful in only 15% of the cases. Fractures with dislocations below 30 degrees, a shortening of less than 5 mm, no rotational displacement or that below 10 degrees, no articular incongruency, and no relevant soft tissue trauma do not need surgery according to our results and should be treated with early mobilization as suggested by the survey. Beyond these limits a primary surgical therapy is justified. The immobilization of metacarpal fractures over a period of more than 3-4 weeks is not necessary.

Adult↗

PubMed1999

Integrating bits and pieces: synapse structure and formation in Drosophila embryos.

During the development of the nervous system, numerous neurons connect to form complex networks. In order to build a functional network each neuron has to establish contacts with appropriate target cells, and at these contacts synapses of the right quality and strength have to be formed. Gaining insight into the mechanisms underlying this complex development is an important step towards a better understanding of how the nervous system is formed and behaviour generated. One model system in which synapse formation can be studied at the morphological, physiological and molecular level is that of the fruitfly Drosophila, and insights gained from Drosophila embryos are reviewed here. The first part of this review deals with the neuromuscular junction as the best-known synaptic contact in Drosophila. It describes: (1) its structure, (2) mechanisms underlying the formation of the neuromuscular cell junction and the arborisation of the presynaptic terminal, and (3) our present understanding of signal-dependent and -independent processes during synapse formation at the neuromuscular junction. The last part of this review deals with the question of how particular neurons can adopt specific synaptic properties, stating as an example the development of the neural lineage of NB7-3, which gives rise to two serotonergic neurons.

Animals↗

PubMed1999

even-skipped determines the dorsal growth of motor axons in Drosophila.

Axon pathfinding and target choice are governed by cell type-specific responses to external cues. Here, we show that in the Drosophila embryo, motorneurons with targets in the dorsal muscle field express the homeobox gene even-skipped and that this expression is necessary and sufficient to direct motor axons into the dorsal muscle field. Previously, it was shown that motorneurons projecting to ventral targets express the LIM homeobox gene islet, which is sufficient to direct axons to the ventral muscle field. Thus, even-skipped complements the function of islet, and together these two genes constitute a bimodal switch regulating axonal growth and directing motor axons to ventral or to dorsal regions of the muscle field.

Animals↗

PubMed1999

Treatment of complete acromioclavicular dislocation: present indications and surgical technique with biodegradable cords.

We report a retrospective study of 48 patients with complete acromioclavicular dislocation (Tossy III). All patients (38 male; 10 female) with an average age of 33.4 years underwent surgery including PDS-augmentation. More than half of the injuries were caused by sport accidents. There were no complications during surgery. 87% of the patients were free of complaints and subjectively very satisfied with the surgical results. By radiological examination we diagnosed a subluxation of the clavicula in 25% of the cases and arthrosis in 17% of the cases. Assessment of subjective complaints, the clinical examination, and the radiological diagnostic according to the Taft Score (0-12 points) resulted in an average value of 10.2 points. The surgical intervention using PDS-cord augmentation in cases of complete acromioclavicular separation is a safe and economic method with a low complication rate. Advantages are possible early-functional treatment, no risk of movement of implants, and avoidance of metal removal.

Absorbable Implants↗

PubMed1999

[The MR tomographic imaging of uncomplicated secondary fracture healing exemplified by the distal radius fracture].

PURPOSE: To investigate the normal sequential MR-pattern of uncomplicated fracture healing within the first 6 weeks. METHODS: In 8 patients with distal radius fracture, 4 by 4 with and without intraarticular fracture, MR examinations were performed 3-4 days after the onset of fracture and after 1, 2, 3, 4, 5 and 6 weeks using the following techniques: T1-weighted spin echo, T2-weighted spin echo, T2 proton density weighted, STIR (short inversion time inversion recovery), T2-weighted turbo spin-echo and fast field echo (FFE, gradient echo). Sequential examinations were analysed regarding 1) the appearance of fracture line, 2) development of bone marrow signal, and 3) signal changes in surrounding soft tissue. Maximum contrast of bone marrow and soft tissue was measured. RESULTS: The fracture line in bone marrow was best detectable in FFE- and T2-weighted spin ech images during the first week and in T2-weighted fast spin echo and proton density-weighted images from the 2nd to 4th week. Beginning in the 5th-6th week an increase of signal intensity in the fracture gap was typical in all sequences except for T1-weighted spin echo. The contrast maximum of bone marrow and soft tissue, best detectable in STIR- and T2-weighted spin echo images, occurred in the 1st-2nd week. CONCLUSIONS: MRI shows a typical time-depending pattern of the fracture line, surrounding bone marrow and soft tissue in normal fracture healing. The degree of injury influences the development of signal changes.

Adult↗

PubMed1999

A role for PS integrins in morphological growth and synaptic function at the postembryonic neuromuscular junction of Drosophila.

A family of three position-specific (PS) integrins are expressed at the Drosophila neuromuscular junction (NMJ): a beta subunit ((betaPS), expressed in both presynaptic and postsynaptic membranes, and two alpha subunits (alphaPS1, alphaPS2), expressed at least in the postsynaptic membrane. PS integrins appear at postembryonic NMJs coincident with the onset of rapid morphological growth and terminal type-specific differentiation, and are restricted to type I synaptic boutons, which mediate fast, excitatory glutamatergic transmission. We show that two distinctive hypomorphic mutant alleles of the beta subunit gene myospheroid (mys(b9) and mys(ts1)), differentially affect betaPS protein expression at the synapse to produce distinctive alterations in NMJ branching, bouton formation, synaptic architecture and the specificity of synapse formation on target cells. The mys(b9) mutation alters betaPS localization to cause a striking reduction in NMJ branching, bouton size/number and the formation of aberrant 'mini-boutons', which may represent a developmentally arrested state. The mys(ts1) mutation strongly reduces betaPS expression to cause the opposite phenotype of excessive synaptic sprouting and morphological growth. NMJ function in these mutant conditions is altered in line with the severity of the morphological aberrations. Consistent with these mutant phenotypes, transgenic overexpression of the betaPS protein with a heat-shock construct or tissue-specific GAL4 drivers causes a reduction in synaptic branching and bouton number. We conclude that betaPS integrin at the postembryonic NMJ is a critical determinant of morphological growth and synaptic specificity. These data provide the first genetic evidence for a functional role of integrins at the postembryonic synapse.

Alleles↗

PubMed1999

Partitioning invertase between a dilute water solution and generated droplets.

Water droplets or mist occur naturally in the air at seashores. These water droplets carry inorganic and organic substances from the sea to the land via the air, creating fertile land in sandy coastal areas (1). The same phenomenon occurs in an air-fluidized bed bioreactor (2). In an air-fluidized bed reactor, proteins can be transferred from the bioreactor semisolid bulk phase to an enriched droplet phase. This protein transfer process (droplet fractionation) can be experimentally simulated by shaking a separatory funnel containing a dilute solution of a given protein, which can be an enzyme like invertase. The created droplets become richer in invertase (protein) than that of the original dilute solution. The droplets can then be coalesced by trapping them and recovering the concentrated protein in the new liquid phase. Typically, in such a droplet fractionation process a collected enzyme can be degraded in its ability to catalyze a chemical reaction. In this article, we explore whether the initial solution pH control variable can be adjusted to minimize the decrease of enzyme activity in this process. The protein droplet recovery problem is one in which the recovered amount of desired protein (enzyme) in the droplet is maximized, subject to the minimization of the enzyme activity loss. The partition coefficient, which is the ratio between the protein concentration in the droplets and the residual solution, is maximized at approx 4.8 and occurs at pH 3.0. Here, the partition coefficient for invertase decreases as the initial solution pH increases, between pH 3.0 and 8.0. Interestingly, the initial solution surface tension seems to be inversely proportional to the partition coefficient. The partition coefficient reaches a maximum value at a surface tension value of approx 63 mN/m at pH 3.0. The enzymatic activity of the initial, the residual, and the droplet solutions all decrease as the bulk solution pH increases. A decrease of enzymatic activity was observed in the residual bulk solution when compared with that in the initial bulk solution at all pH levels. Also, up to 90% of the invertase activity was lost in the droplets when compared to the initial bulk solution.

Journal Article↗

PubMed1999

Preserving the activity of cellulase in a batch foam fractionation process.

Foam fractionation is one of the low operating-cost techniques for removing proteins from a dilute solution. The initial bulk solution pH and air superficial velocity play an important role in the foam-fractionation process. Denaturation of proteins (enzymes) can occur, however, during the foam-fractionation process from the shear forces resulting from bursting air bubbles. At the extreme bulk solution pHs (lower than 3.0 and higher than 10.0), the enzymatic activity of cellulase in the foamate phase drops significantly. Within these two pH bounds an increase in the air superficial velocity, V0, and a decrease in the bulk solution pH leads to a decrease in the separation ratio (SR), defined as the ratio of the protein concentration in the foamate to the protein concentration in the residue. On the other hand, an increase in V0 provides a higher foamate-protein recovery. The process efficiency is defined as the product of foamate-protein recovery times the SR times the cellulase activity. The optimal operating condition of the cellulase foam-fractionation process is taken into account at the maximum value of the process efficiency. In this study, that optimal condition is at an air superficial velocity of 32 cm/min and a bulk-solution pH of 10.0. At this condition, the recovered foamate is about 80% of the original protein mass, the SR is about 12, and the enzymatic activity is about 60% of the original cellulase activity.

Journal Article↗

PubMed1998

The kakapo mutation affects terminal arborization and central dendritic sprouting of Drosophila motorneurons.

The lethal mutation l(2)CA4 causes specific defects in local growth of neuronal processes. We uncovered four alleles of l(2)CA4 and mapped it to bands 50A-C on the polytene chromosomes and found it to be allelic to kakapo (. Genetics. 146:275- 285). In embryos carrying our kakapo mutant alleles, motorneurons form correct nerve branches, showing that long distance growth of neuronal processes is unaffected. However, neuromuscular junctions (NMJs) fail to form normal local arbors on their target muscles and are significantly reduced in size. In agreement with this finding, antibodies against kakapo (Gregory and Brown. 1998. J. Cell Biol. 143:1271-1282) detect a specific epitope at all or most Drosophila NMJs. Within the central nervous system of kakapo mutant embryos, neuronal dendrites of the RP3 motorneuron form at correct positions, but are significantly reduced in size. At the subcellular level we demonstrate two phenotypes potentially responsible for the defects in neuronal branching: first, transmembrane proteins, which can play important roles in neuronal growth regulation, are incorrectly localized along neuronal processes. Second, microtubules play an important role in neuronal growth, and kakapo appears to be required for their organization in certain ectodermal cells: On the one hand, kakapo mutant embryos exhibit impaired microtubule organization within epidermal cells leading to detachment of muscles from the cuticle. On the other, a specific type of sensory neuron (scolopidial neurons) shows defects in microtubule organization and detaches from its support cells.

Alleles↗

PubMed1998

Absence of PS integrins or laminin A affects extracellular adhesion, but not intracellular assembly, of hemiadherens and neuromuscular junctions in Drosophila embryos.

We have examined the role of integrins in the formation of the cell junctions that connect muscles to epidermis (muscle attachments) and muscles to neurons (neuromuscular junctions). To this end we have analyzed muscle attachments and neuromuscular junctions ultrastructurally in single or double mutant Drosophila embryos lacking PS1 integrin (alphaPS1betaPS), PS2 integrin (alphaPS2betaPS), and/or their potential extracellular ligand laminin A. At the muscle attachments PS integrins are essential for the adhesion of hemiadherens junctions (HAJs) to extracellular matrix, but not for their intracellular link to the cytoskeleton. The PS2 integrin is only expressed in the muscles, but it is essential for the adhesion of muscle and epidermal HAJs to electron dense extracellular matrix. It is also required for adhesion of muscle HAJs to a less electron dense form of extracellular matrix, the basement membrane. The PS1 integrin is expressed in epidermal cells and can mediate adhesion of the epidermal HAJs to the basement membrane. The ligands involved in adhesion mediated by both PS integrins seem distinct because adhesion mediated by PS1 appears to require the extracellular matrix component laminin A, while adhesion mediated by PS2 integrin does not. At neuromuscular junctions the formation of functional synapses occurs normally in embryos lacking PS integrins and/or laminin A, but the extent of contact between neuronal and muscle surfaces is altered significantly. We suggest that neuromuscular contact in part requires basement membrane adhesion to the general muscle surface, and this form of adhesion is completely abolished in the absence of laminin A.

Animals↗

PubMed1998

The muscleblind gene participates in the organization of Z-bands and epidermal attachments of Drosophila muscles and is regulated by Dmef2.

We report the embryonic phenotype of muscleblind (mbl), a recently described Drosophila gene involved in terminal differentiation of adult ommatidia. mbl is a nuclear protein expressed late in the embryo in pharyngeal, visceral, and somatic muscles, the ventral nerve cord, and the larval photoreceptor system. All three mbl alleles studied exhibit a lethal phenotype and die as stage 17 embryos or first instar larvae. These larvae are partially paralyzed, show a characteristically contracted abdomen, and lack striation of muscles. Our analysis of the somatic musculature shows that the pattern of muscles is established correctly, and they form morphologically normal synapses. Ultrastructural analysis, however, reveals two defects in the terminal differentiation of the muscles: inability to differentiate Z-bands in the sarcomeric apparatus and reduction of extracellular tendon matrix at attachment sites to the epidermis. Failure to differentiate both structures could explain the partial paralysis and contracted abdomen phenotype. Analysis of mbl expression in embryos that are either mutant for Dmef2 or ectopically express Dmef2 places mbl downstream of Dmef2 function in the myogenic differentiation program. mbl, therefore, may act as a critical element in the execution of two Dmef2-dependent processes in the terminal differentiation of muscles.

Animals↗

PubMed1998

ROP, the Drosophila Sec1 homolog, interacts with syntaxin and regulates neurotransmitter release in a dosage-dependent manner.

The Sec1 family of proteins is thought to function in both non-neuronal and neuronal secretion, although the precise role of this protein family has not been defined. Here, we study the function of ROP, the Drosophila Sec1 homolog, in neurotransmitter release. Electrophysiological analyses of transgenic lines overexpressing ROP and syntaxin, a presynaptic membrane protein, indicate that ROP interacts with syntaxin in vivo. Characterization of four point mutations in ROP shows that they fall into two phenotypic classes. Two mutations cause a dramatic reduction in both evoked and spontaneous neurotransmitter release. In contrast, the other two mutations reveal an increase in evoked neurotransmission. Our data further show that neurotransmission is highly sensitive to the levels of ROP function. Studies on heterozygote animals indicate that half the amount of wild-type ROP results in a dramatic decrease in evoked and spontaneous exocytosis. Taken together, these results suggest that ROP interacts with syntaxin in vivo and is a rate-limiting regulator of exocytosis that performs both positive and inhibitory functions in neurotransmission.

Animals↗