Biomedical subjects

J Stulík

Publications and source records attributed to J Stulík.

At least 37 records · Page 2Linked to original sources

PubMed2002

[Primary osteosynthesis of the odontoid process: a multicenter study].

PURPOSE OF THE STUDY: Direct osteosynthesis is a method of choice for the treatment of odontoid process fractures. It is based on insertion, from the anterior approach, of one or two screws from the C2 body into the apex of the odontoid across the fracture line. The tensile action of screws results in compression of fragments and stabilization of the fracture. The aim of the study was to evaluate a group of patients treated by this method and to compare our results with those reported in the foreign literature. MATERIAL: A total of 99 patients were treated by direct osteosynthesis of the odontoid in the departments involved in the study between 1994 and 2001. METHODS: Patients indicated for this surgery were those with fractures of type II according to Anderson and D'Alonzo and those with type III fractures but only when the fracture line went across the articulation surfaces of C1-C2, when closed reduction was not possible or the patients were not indicated for halo fixation. Direct osteosynthesis was not applied to fractures with comminution at the base of the odontoid, irreducible fractures, odontoid fractures combined with dislocated fractures of the atlas or pathological fractures. Severe kyphosis of the cervical spine or a large thoracic cage was also regarded as a contraindication. RESULTS: All the 99 patients were followed up from 3 up to 102 months, with an average of 28.5 months; only in seven patients, the follow-up period was shorter than 6 months. The most frequent subjective complaint was a painful operation wound. This usually resolved within two weeks of surgery. Except for four patients, alle were satisfied with the outcome. Type II fractures were diagnosed in 84 and type III fractures in 15 cases. A total of 174 screws were inserted into the odontoid processes of 99 patients. A single screw was used in 25 and two screws in 73 patients. In one case, three screws had to be inserted. Screw lenght ranged from 36 to 44 mm, diameter was 40.9 mm. Three months after surgery, X-ray examination, both in flexion and extension, did not reveal any instability in any of the patients. No morphological change in the C2-C3 intervertebral space was observed Of 92 (92.9%) paitents under longterm follow-up, 84 (91.3%) showed complete healing of the fracture, three died and five patients eventually developed pseudoarthrosis, which was due to a broken screw in three of them. This condition was treated by dorsal fixation of C1-C2 according to Magerl or by one of the dorsal cerclage techniques. The group was free of any perioperative complications related to the anterior approach or injury to nerve structures by screws. DISCUSSION: The most frequent subjective complaint was a painful operation wound. Treatment of odontoid fractures varies according to the type of injury, bone quality and also practice at each department. Type II injuries are highly unstable and, because of the small fracture surface, their healing ability is much lower than in type III fractures. Previously, most of the patients with odontoid injuries were treated conservatively by immobilization in a plaster cast or a brace or, later, by a halo device. In the long term, however, they showed a high proportion of pseudoarthroses (10 to 100%). Direct osteosynthesis of the odontoid by screws permits the maintenance of rotation of the C1-C2 mobile segment. We followed the scheme of indications used abroad but did not perform osteosynthesis to correct pseudoarthrosis. The number of osteosyntheses healed (91.3%) was also in agreement with the literature data. Earlier, we used two screws for all types of fractures. Recently, we have preferred insertion of a single screw in type II and III fractures in narrow odontoids. In the later, there is no danger of rotational dislocation during screw insertion; to insert one screw from the centre of the C2 base is easy and speeds up the procedure. However, in displaced type II and type II T fractures, two screws are a necessity. Similarly to other authors, we recorded a slight limitation of cervical spine rotation in patients at long-term follow-up, particularly in elderly subjects with advanced osteochondrosis. No complications leading to deterioration of the patient's state were recorded. CONCLUSIONS: Direct osteosynthesis is a method of choice for most of the type II and indicated cases of type III fractures of the odontoid process of the axis. This surgical procedure facilitates restoration of anatomical conditions of the spine and its immediate stability. Consequently, patients can be readily mobilized and rehabilitated.

Adolescent↗

PubMed2002

[Replacement of the vertebral body with an expansion implant (Synex)].

PURPOSE OF THE STUDY: This paper describes replacement of the vertebral body with the expansion implant Synex. Usually, autologous bone graft is used to replace the vertebral body. In patients with bone cancer or multiple injuries to the spine, cement filling is preferred whereas, in other indicated cases, implants are inserted, of which Harms' titanium cage has been the most common one. However, this needs filling with a large amount of bone tissue and it is often difficult to adjust its size into the space available. Telescopic devices, on the other hand, are easier to implant and their application requires only a minimum amount of autologous bone tissue. MATERIAL: In the period from January 2000 to June 2001, we used telescopic implants Synex to replace vertebral bodies in 34 patients. Indications for treatment were: vertebral fractures in 14, post-traumatic kyphosis in six, vertebral metastatic tumours in eight and a primary tumour in six patients. METHODS: In 25 cases, the vertebral body replacement was completed by posterior stabilization using internal fixation and, in nine cases, by anterior stabilization with a Ventrofix fixator. In 32 patients, the implant was inserted from the anterior approach and, in two, from the posterior approach following complete spondylectomy. RESULTS: The L1 vertebra was replaced most frequently (nine patients), then T 12 (seven patients) and L2 (six patients). For treatment of fresh fractures, the Synex implant was used in 14 cases. Of these one was inserted from the posterior approach in the L1 region where trauma had caused severe injury to the spinal cord. In spinal tumours. Synex was used in 14 patients, i.e., in six with diagnosed plasmacytoma, in two with metastatic dissemination from prostate carcinoma, in four with vertebral metastases from breast cancer and in two patients with non-differentiated metastases. The anterior approach was performed by conventional thoracotomy or combined thoracotomy and lumbotomy in 20 patients and a less invasive retroperitoneal approach was used in 12 patients. One patient died of multiple metastases at 7 months after surgery and one patient had relapse of a local tumour resulting in paraparesis that required a repeat decompression of the spinal canal. The operation took 1 h and 50 min when the anterior approach and anterior stabilization with a Ventrofix fixator were used; the operation lasted from 3 h 20 min to 6 h 10 min when complementary posterior stabilization was involved. The patients were followed up for 2 to 24 months. No failure of the implant in terms of migration, change in position or penetration into adjacent vertebral bodies occurred. DISCUSSION: The replacement of a vertebral body has conventionally been performed with the use of a massive bone graft. However, collection of an autologous bone graft large enough to suit this purpose is not always possible. Complications at the donor site have been described. A homologous bone graft carries a risk of disease transmission and the reconstruction ability of a massive graft has not been confirmed for certain. Cement filling augmented with Kirschner's wires is usually used in cancer patients. Titanium cages require application of a large amount of spongiose bone tissue into their interior. Consequently, bone in the centre fails to remodel. A sharp edge of the mesh may induce migration of the cage towards the vertebral body and failure of the implant. Mechanical failure and collapse of cages have also been described. Telescopic cylindrical implants, on the other hand, need only a small amount of spongiose bone tissue to fill. They can be adapted directly to the implantation site by means of a special distractor and, therefore, before adjusting its final length, the exact position and orientation of the implant can be achieved in the space prepared. This facilitates close contact with the endplates of adjacent vertebral bodies and the development of osteointegration. The use of telescopic implants enabled us to avoid the force that is often necessary to apply during insertion of Harms' cages in the patients whose spines had already been stabilized with posterior fixation or to avoid the need of a triple surgical procedure in order to achieve better stability of the implant. In two patients, Synex was inserted from a non-standard posterior approach. Indications for Synex implantation should be evaluated in view of disease prognosis in each patient. If only limited survival is expected, cement filling with K-wires should be preferred. CONCLUSIONS: Synex is a sophisticated implant to replace severely damaged vertebral bodies regardless of the nature of lesion. Its application required additional stabilization by either posterior or anterior fixation (internal transpedicular fixator and Ventrofix or Kaneda, respectively). Its use is indicated in post-traumatic defects of vertebrae in acute or poorly healed scervical.

Bone Transplantation↗

PubMed2002

[Combined method of treating dislocated fractures of the calcaneus].

PURPOSE OF THE STUDY: The treatment of dislocated intraarticular fractures of calcaneus is still an unsolved chapter of traumatology. Our own poor long-term results of a purely conservative procedure has led us to develop a combined method of a direct and indirect reduction of calcaneus with the subsequent stabilization which we have been using since 1994. The basic principle of the method consists in the combination of a direct reduction of dislocated fragments of articular surfaces and indirect of calcaneus as a whole with the subsequent transfixation of calcaneus by K-wires. MATERIAL: In the period of 1994-2001 we have treated in total 261 patients with 302 fractures of calcaneus, of which 218 were men (83.5%) and 43 women (16.5%) in the age range from 13 to 82 years (average age 45.1 years). By our own surgical method we treated 213 patients with 248 fractures, of which 180 were men (84.5%) and 33 women (15.5%) in the age range of 13-79 years (average age 44.8 years). Conservatively handled were 54 fracture in 48 patients. In the long-term followed up group we included 160 patients with 187 fractures operated on by identical method observing a minimum two-year interval after the operation. Of them 134 were men (83.8%) and 26 women (16.2%) in the age range of 15-75 years (average age 44.3 years). Of 248 operated on fractures of calcaneus 159 were joint-depression type (64.1%), 61 tongue type (24.6%) and 28 comminuted (11.3%) types of fractures. The time interval between the injury and operation ranged between 6 hours and 3 weeks (average 18.5 hours), however most patients we operated on on the day of injury. METHODS: The basic principle of the method was a combination of a direct reduction of dislocated fragments of articular surfaces and indirect reduction of the calcaneus as a whole with a subsequent transfixation of the calcaneus by K-wires. After draping and setting of the image intensifier we proceeded to the actual surgery and divided the whole procedure of reduction and subsequent stabilization into 4 phases: Phase I--traction Phase II--elevation Phase III--compression Phase IV--transfixation In the first traction phase we inserted K-wire transversely through the distal-posterior edge of the calcaneus and connected it to the U-handle in order to achieve a proper effect of the traction. By traction distally along the long axis of the limb we restored the height and length of the calcaneus. Simultaneous pendular movements in eversion/inversion direction released the fragments and considerably facilitated the subsequent reduction. In the second, elevation phase the procedure differed according to individual types of fracture. In joint-depression type we inserted a blunt Steinmann pin or better a curved transpedicular elevator under the dislocated posterior articular surface through skin plantar stab incision and an ever-present primary transverse line of the fracture. In the tongue type of fracture we first introduced in the second phase a thick K-wire (3 mm) along the long axis of the tongue fragment and reduce the fragment by leverage elevation with a simultaneous continuous U-handle traction. After obtaining the desirable position we fixed the fragment by a K-wire vertically introduced from the upper part of calcaneal tuberosity (anteriorly and laterally from the origin of the Achilles tendon) towards the planta. If the medial part of the posterior articular surface remained dislocated we finished its reduction by means of elevator as in the preceding type of the fracture. The third and fourth phases of the surgical procedure was again identical for both types of fractures. In the third, compression phase we performed manually lateral compression of fragments under permanent traction. In the fourth, transfixation phase we first transfixed the reduced position of fragments formed by the primary longitudinal and transverse line. Under continuous radiograph checking in the lateral projection we inserted K-wires gradually from the lateral aspect, about 1.5 cm beneath the lateral malleolus into sustentacular fragment. Additional, mostly 2 K-wires we inserted under continuous traction and counter-traction along the long axis of the calcaneus and as the last step we drilled K-wires from the plantar side from calcaneal tuberosity into the fragments of the posterior articular surface. RESULTS: In the group of long-term followed up patients we evaluated the Creighton-Nebraska Health Foundation score in 160 patients who were on average 43.4 months after operation. The obtained values of the score ranged between 63-100 points with the average of 83.9 points. In 27 patients the result was very good (16.9%), in 89 patients good (55.6%), in 24 patients fair (15.0%) and in 20 patients the result was poor (12.5%). DISCUSSION: The first results of our method of 1998 have proved that it is necessary to combine the basic procedures of direct reduction of joint fragments, i.e. restoration of the calcaneus as a whole, namely in a strictly recommended sequence with a subsequent transfixation by K-wires. We do not use surgical procedures with open reduction and internal fixation (ORIF) differing mutually by the chosen surgical approach and the type of internal fixation used. As compared to ORIF our method has a significantly broader indication range. It can be used for the operation of patients regardless of the age, presence of associated diseases (diabetes, vascular affection) or local affection (a marked oedema, haematoma, non-infected skin blisters). The comparison of the general outcomes of the treatment in our group of patients, 72.5% of excellent and good results and only 12.5 of poor results, corresponds with the values of equally extensive foreign group of patients. In addition, as concerns the number of the incidence of deep infects in closed fractures (0.8%) the values in our group are in comparison several times lower and amputation of the limb was not necessary in any of the cases. It should be also noted that our group of patients has only a minimal indication limitation and operated on are also risk patients who never get in "filtered" groups treated by open method. Therefore it may be stated that the general results are in case of our method markedly better. CONCLUSION: The proposed method requires neither a specialist nor any expensive technical equipment and as a result it may be used both at orthopaedic and surgical departments and due to its undemanding nature and short hospitalization it is also very acceptable from the economic viewpoint.

Adolescent↗

PubMed2002

[Use of bioceramics in the treatment of fractures of the thoraco-lumbar spine].

PURPOSE OF THE STUDY: The primary reduction and stabilization of all types of injury to the thoracolumbar spine is currently performed from the posterior approach by an internal, transpedicular fixator. The exceptions are type A fractures, according to the AO classification, that can primarily be treated from the anterior approach. The aim of the study was to assess the effect of BAS-O bioceramic granules, inserted by transpedicular approach, on the development of post-operative kyphosis of the segments injured. MATERIAL: Between 1997 and 2001 we treated anterior spinal column fractures, using bioceramic granules in combination with an internal fixator, in 53 patients (40 men, 13 women: age 26 to 69 years; average, 42.3 years) at the Department of Orthopedics and Traumatology of the Third Faculty of Medicine and FNKV in Prague. However, only 42 patients (33 men, 9 women; age 28 to 67 years; average, 41.6 years) who had had the metal fixator removed more than six months previously were included in the study. The fixator was removed in the range of 10 to 24 months after the primary operation. The sample was divided into two groups; one with bioceramic material inserted in the body of the damaged vertebra (20 patients) and the other bioceramic granules implanted in both the vertebral body and the intervertebral space (22 patients). METHODS: In all the patients, the standard procedure included transpedicular application of Schanz's screws and an USS FS fixator (Synthes). If necessary, distraction of the segment injured and the correction of lordosis were carried out. Further transpedicular procedures to treat the anterior spinal column were as follows: If the vertebral body was injured without destruction to the intervertebral disc, a bent elevator was inserted through the vertebral foramen and the intervertebral joints were reduced. At the same time, a cavity for application of bioceramic granules was created. These were inserted, using a funnel and a pusher, in the anterior part of the injured body. If the fracture involved a destroyed intervertebral disc, the disc was removed, the vertebral end plate of the adjacent vertebra was perforated and bioceramic granules were inserted in both the vertebral body and intervertebral space. Subsequently, spongeous bone grafts were harvested from the ilium and massively applied to the previously decorticated transverse and articular processes. RESULTS: In the group of patients who had bioceramic granules inserted in only the vertebral body, the kyphotic angle was on average 10.36 degrees after injury and 2.86 degrees after surgery; therefore, a correction by 13.22 degrees was achieved. At 3 and 6 months after surgery, the kyphotic angle was 2.71 degrees and 2.68 degrees, respectively; at 3 and 6 months after fixator removal, it was 0.67 degree in both instances. In the group of patients with bioceramic granules implanted in both the vertebral body and intervertebral space, the kyphotic angle was on average 9.16 degrees after injury and 4.26 degrees after, surgery; therefore, a correction by 13.43 degrees was achieved. At 3 and 6 months after surgery, the kyphotic angle was -4.11 degrees and -4.00 degrees, respectively; at 3 and 6 months after fixator removal, it was 2.38 degrees and 2.44 degrees, respectively. DISCUSSION: Our results revealed differences between the patients who had bioceramic granules inserted in only the vertebral body and those who had them also in the intervertebral space. At 6 months after surgery, the first group showed the loss of correction per two segments to be 3.53 degrees on the average, whereas the second group had a loss of 6.70 degrees, i.e., twice as high. This may be explained by a more serious damage to the intervertebral disc in the latter group. Only small differences between the groups were found in the kyphotic angle at both 6 months after surgery and 3 to 6 months after fixator removal. This implied that, in both groups, the loss of correction occurred in the period up to 3 months after removal of the fixator. CONCLUSIONS: Bioceramic granules provide material for replacement of osseous tissue in the body of the vertebra as well as conditions necessary for bone restructuring. The loss of correction per segment is lower by about half in patients treated with bioceramic granules than in those who received a spongeous bone grafts.

Adult↗

PubMed2001

Proteome study of colorectal carcinogenesis.

Development of cancer is a complex process involving multiple changes in gene expression. To unravel these alterations, a proteome approach aimed at the identification of qualitative and quantitative changes in protein composition, including their post-translational modifications, attracts great attention. Our study was focused on the identification of proteins whose amount is altered in the course of malignant transformation of colon mucosa. Proteins extracted from tissue specimens or cell lysates were separated by two-dimensional gel electrophoresis (2-DE). Comparative analyses of 2-DE protein patterns were done using computerized image analysis. Selected proteins exhibiting statistically significant abundance alterations comparing healthy and diseased tissues were identified by mass spectrometry. Globally, we have found 57 proteins that exhibited either a significant decrease or increase in amount in pathological tissues, and 18 of these were annotated by mass spectrometry. The alterations in the expression of nine proteins were common for both precancerous and neoplastic tissues suggesting their role in colon tumorigenesis. The epithelial origin of all identified spots was checked in two cell lines Caco-2 and DLD-1 originating from well-differentiated and poorly differentiated colon carcinoma, respectively.

Adenocarcinoma↗

PubMed2001

Construction of a Francisella tularensis two-dimensional electrophoresis protein database.

We have started the construction of a two-dimensional database of the proteome of Francisella tularensis, a bacterium that is responsible for the highly pathogenic disease tularemia. The genome of this intracellular pathogen is not completely sequenced yet and, currently, information about only 66 proteins is available from NCBI database. We have analyzed the F. tularensis live vaccine strain by two-dimensional gel electrophoresis with immobilized pH 3-10 gradient in the first dimension and 9-16% gradient or tricine SDS-PAGE in the second dimension. In both cases about 2000 spots were detected. Furthermore, we compared the protein pattern of the nonvirulent F. tularensis live vaccine strain with protein profiles of two wild type clinical isolates and more than 50 differentially expressed proteins were counted. The separated proteins are going to be identified by peptide mass fingerprinting. However, due to the lack of complete genome sequence data only eight proteins were unambiguously identified. Among them, acid phosphatase and the most basic isoform of a hypothetical 23 kDa protein are characteristic only for virulent strains.

Bacterial Proteins↗

PubMed2001

[Tension cerclage using scews--a forgotten technique?].

Authors deal with the screw-wire technique described by Brunner and B. G. Weber in 1981. The principle of the technique consists in passing a wire loop around the heads of divergently driven screws. After tightening the screws the wire loop is put under tension which results in interfragmental compression. Mostly used for this purpose are 4.5 mm cortical screws but, if need be, any screw from a standard or small set of instruments is applicable. This type of fixation they use mainly for acetabulum fractures, diastasis of symphysis or for the stabilization of knee osteotomies.

Acetabulum↗

PubMed2001

[Treatment of thoracolumbar spinal fractures using internal fixators (evaluation of 120 cases)].

PURPOSE OF THE STUDY: A group of 120 patients was evaluated with acute injury of Th-L spine who were operated on from posterior approach with the use of Dick AO fixator or USS fixator in the period of 1991-1996. MATERIAL: In the period of 1991-1996, 128 patients were operated on with the acute injury of Th-L spine. Evaluated were 120 patients: 81 men (67.5%) of average age 40.1 years (age range, 16-74 years) and 39 women (32.5%) of average age 35.4 years (age range, 15-66 years). Posterior stabilization using the internal fixator was combined with posterolateral or posterior interarticular fusion. In comminuted fractures of the vertebral body transpedicular cancellous bone grafting was performed and starting for 1995 also intercorporal transpedicular fusion. METHODS: The evaluation covered location and type of fractures, injury of other levels of the spine, other associated injuries, neurological symptomatology prior to the surgery and 3 months after the surgery, duration of surgery and of image intensifier exposure, peroperative and early postoperative complications, radiograph evaluation of the degree of reduction, the loss of correction after 12 months and after the hardware removal. RESULTS: Evaluated were 120 patients. Average interval after the surgery at the time of the evaluation was 43.7 months (range, 24-93 months). The injury was caused in 82 cases (68.3%) by a fall from height, in 26 cases (21.7%) by a car accident and in 12 cases (10%) by another cause. Associated injuries included: calcaneus fracture 18 times, the fracture of ankle 4 times, pelvis fracture 4 times, other fractures 18 times. In 89 cases the injury involved only one vertebra, i.e. one level, 31 cases were multi-level injuries. The angle of the kyphosis of vertebral body was on average 21.8 degrees. Patients with neurological symptoms were on average operated on 2-12 hours after the admission, other patients were operated on in the interval between 5 hours and 7 days. Bilateral transpedicular cancellous bone grafting was performed in 106 affected vertebrae, the canal was revised in 27 injured vertebrae. The surgery took on average 172 minutes (range, 62-430 minutes). Image intensifier exposure lasted on average 2.4 minutes. Average blood loss was peroperatively estimated at 725 ml, postoperatively on average at 815 ml. Postoperative complications: healing of the wound per secundam 6 times, thromboembolic complication in 9 cases--never fatal, pneumonia 3 times, uroinfection 5 times. The total number of these complications was about 8%. Vertebral body was completely restored in 72 cases, incompletely in 34 cases (up to 5 degrees deficit). The correction of kyphosis achieved was on average 6.5 degrees. After 12 months the correction loss with the fixator in place was on average 2.5 degrees. After the removal of the fixator the average correction loss was 5.5 degrees. In 2 cases there occurred a significant re-dislocation which required revision surgery. Transpedicular screws broke 6 times in the total of 4 patients, fixator loosened in 2 cases. DISCUSSION: Surgical treatment of thoracolumbar spine is indicated in unstable injuries and all injuries with neurological lesion and a proved compression of spinal canal. Decompression of the spinal canal is often achieved by reduction, lordotization and distraction. In other cases revision of the canal is indicated. Stabilization is performed by means of a transpedicular internal fixator. Part of the surgery is posterior fusion. The defect in the reduced body is treated by transpedicular cancellous bone grafting. The affected inervertebral disc must be treated by intersomatic fusion. Anterior operation is indicated as a complementary one. CONCLUSIONS: 1. Absolute majority of injuries of thoracolumbar spine may be treated from the posterior approach. In this it is necessary to restore the stability of the anterior column. An indispensable part of the operation is a flawless posterior fusion. 2. Some types of injuries require a complementary anterior approach in the first or second step. This applies mainly to the injuries with a defect of anterior column evaluated on the basis of classification after Gaines. 3. A primary isolated anterior approach to the treatment of the injury of Th-L spine is not in our view suitable as a routine.

Adolescent↗

PubMed2000

Differential expression of the Ca2+ binding S100A6 protein in normal, preneoplastic and neoplastic colon mucosa.

The expression of calcium-binding protein S100A6 was investigated in normal colon tissue, in colon adenomas and in colorectal carcinomas. Using an immunoblotting approach we detected four S100A6 variants with Mwt of 10 kDa and pI of 5.05 (isoform I), 5.15 (isoform II), 5.23 (isoform III) and 5.32 (isoform IV) that were differentially expressed in the analysed samples. The quantitative examination of S100A6 variant expression in 25 pairs of colorectal carcinoma and matched control mucosa proved a statistically significant increased abundance of S100A6 isoforms I (P = 0.004) and III (P = 0.025) in malignant tissue, and conversely, an increased level of S100A6 isoform IV in healthy tissue (P = 0.022). The expression of isoforms I and III and the loss of isoform IV were also observed in colon cancer cell lines. In addition, the immunohistochemical study of 16 primary colorectal carcinomas revealed both in the non-paired Student t-test and in the Mann Whitney test the statistically significant accumulation of S100A6 protein (P < 0.001) in the invasive margin of the tumour. The immunohistochemical analysis of S100A6 protein in polyps differing in clinical severity gave a strong staining that was maximal in dysplastic lesions. Thus, our results indicate a possible, statistically significant correlation (non-paired Student t-test P = 0.036) between S100A6 expression and colon carcinoma progression.

Adenoma↗

PubMed2000

Protein changes in HL60 leukemia cells associated with 5-aminolevulinic acid-based photodynamic therapy. Early effects on endoplasmic reticulum chaperones.

Using two-dimensional electrophoresis we investigated the effect of 5-aminolevulinic acid (ALA)-based photodynamic therapy (PDT; induction with 1 mM ALA for 4 h followed by blue light dose of 18 J/cm2) on the protein expression in HL60 leukemia cells. ALA-PDT resulted in extensive qualitative and quantitative changes in the protein pattern of HL60 cell lysates. Of more than 1350 protein spots recognized on the protein maps of ALA-induced cells, seven proteins were enhanced and 17 suppressed following irradiation. Three of these, calreticulin precursor, p58 microsomal protein (ERp57) and protein disulfide isomerase (p55) have been identified by matrix-assisted laser desorption and ionization-mass spectrometry and the pI/molecular weight parameters of the affected proteins were estimated by computer analysis. The findings suggest participation of endoplasmic reticulum Ca(2+)-binding chaperones and/or Ca2+ signaling in ALA-PDT mediated cytotoxicity.

Aminolevulinic Acid↗

PubMed2000

Electrophoretic characterization of a gamma-1-heavy chain disease.

This report describes a new case of gamma-1-heavy chain disease found in a woman with malignant lymphoproliferative disease. The patient's serum and urine containing gamma-1-heavy chains were analyzed using different electrophoretic approaches, especially two-dimensional electrophoresis and immunoblotting analysis. In a serum sample, five sets of gamma-1-heavy chain spots differing in molecular weight with acidic pI values and one set of more basic gamma-1-heavy chain spots were found. The major group of spots exhibited molecular weight in the range from 29 to 39 kDa. Examination of urine sample proved the presence of the more basic set of gamma-1-heavy chain spots and two acidic groups, including 29 to 39 kDa set.

Aged↗

PubMed1999

Protein abundance alterations in matched sets of macroscopically normal colon mucosa and colorectal carcinoma.

Our current results, aimed at the detection of protein abundance alterations that could be associated with the process of colon tumorigenesis, are summarized. The matched sets of macroscopically normal colon mucosa and colorectal carcinoma were examined by a one- or two-dimensional electrophoretic approach and proteins were identified using immunoblotting or mass spectrometry. The following results were observed: The levels of liver fatty acid-binding protein, actin-binding protein/smooth muscle protein 22-alpha and cyclooxygenase 2 were downregulated in colorectal carcinoma compared to normal colon mucosa. Conversely, the expression of a novel variant of heat shock protein70 and several members of the S100 protein family of calcium-binding proteins (two isoforms of S100A9, S100A8, S100A11 and S100A6) were upregulated in transformed colon mucosa. Despite the variations of the levels of expression of given protein among analyzed samples, all quantitative changes were found to be statistically significant (Mann-Whitney test assuming p < or = 0.05). We conclude that the proteomic approach is useful for the study of complex biological events underlying the process of colorectal tumorigenesis.

Colon↗

PubMed1999

Production of stress-inducible form of heat-shock protein 70 in mouse peritoneal adherent cells after in vivo infection by Francisella tularensis.

Heat-shock proteins (hsp) are ubiquitously produced molecules which participate in the protection of cells from environmental perturbation. Moreover, the members of the heat-shock protein 60 (hsp60) and 70 (hsp70) families play an important role in pathogen-host interactions. We studied in vivo production of the 70-kDa heat-shock proteins in the extract of peritoneal exudate cells (PEC) from mice injected intraperitoneally with an attenuated vaccine strain (LVS) of Francisella tularensis. We found a differential production of a highly stress-inducible member of the hsp70 family, designated hsp72, in three inbred strains of mice exhibiting either resistance or susceptibility to F. tularensis LVS infection. Whereas in tularemia-resistant mice hsp72 was even expressed in PEC without injection of bacteria and its production further increased on day 3 and slowly declined on days 5 and 7 after injection, in susceptible mice hsp72 production was highly inducible and restricted only to day 3 after in vivo infection. Further analysis of hsp72 expression revealed intracellular hsp72 accumulation and its preferential production by peritoneal adherent cells.

Animals↗

PubMed1998

Natural resistance to intracellular parasites: a study by two-dimensional gel electrophoresis coupled with multivariate analysis.

Natural resistance to Mycobacterium bovis bacillus Calmette-Guérin (BCG) is determined by the Bcg gene (Nramp1), which is exclusively expressed by mature macrophages. The Nramp1 gene is a dominant autosomal gene that has two allelic forms; r confers resistance and s confers susceptibility to infection with intracellular pathogen. Although the wide range of pleiotropic immunological effects of the Nramp1 gene has been described, the exact mechanism of its action remains elusive. In this study we searched for differentially expressed proteins that might provide clues in the studies on Nramp1 gene function. We performed two-dimensional gel electrophoresis of cellular proteins prepared from a B10R macrophage line derived from mice carrying the r allele of the Nramp1 gene, B10S macrophages carrying the s allele, and B10R-Rb macrophages transfected with Nramp1-ribozyme. The classification of protein patterns and selection of distinct proteins characteristic of r or s allele-carrying macrophages was performed using the principal component analysis. We found differential expression of four proteins with the following isoelectric point/molecular weight (pI/Mr) in B10R macrophages compared to B10S and B10R-Rb macrophages: 6.6/25, 7.0/22, 9.1/31.5, and 5.3/8.5. The protein 7.0/22 has been identified as Mn-superoxide dismutase and the best candidate for protein p6.6/25 seems to be Bcl-2 according to the immunoblot analysis. When the splenic macrophages carrying the r or s allele were analyzed, the changes in relative abundance for proteins 6.6/25 and p7.0/22 were satisfactorily reproduced. Overall, the two identified proteins are important in the regulation of intracellular redox balance and the regulation of apoptosis in macrophages, respectively. Our findings may suggest their possible biological role in the innate immunity against intracellular pathogens.

Animals↗

PubMed1998

[Our method of treatment of dislocated fractures of the heel bone (preliminary report)].

The authors present first experience with their method of treatment of displaced fractures of calcaneus. The method is based on the combination of direct and indirect reposition of fragments and exact fixation with K-wires. Axial and AP X-ray and 2 plane CT scan are indispensible conditions for operation planning. The procedure can be divided into 4 phases: traction, elevation, compression, transfixation. Up till now 65 patients have been treated with this method. 20 patients with 24 fractures were evaluated according to the Creighton-Nebraska Health Foundation score in the interval 24 months after the operation. The average value of the score was 96.6 points, 16 patients were evaluated as excellent, in 2 patients the result was good.

Adolescent↗

PubMed1997

Overexpression of calcium-binding protein calgranulin B in colonic mucosal diseases.

Subtractive two-dimensional gel electrophoresis (2-DE) has been used for the study of the protein patterns of the normal colonic mucosa and the specimens collected from patients diagnosed for inflammatory bowel disease (IBD), colonic polyps and colorectal cancer. We found a 13 kDa protein that was detected in five of seven adenomas and in 13 of 15 colorectal carcinomas while it was absent or only slightly expressed in normal colonic mucosa. Furthermore, this protein occurred in all specimens collected from patients suffering from IBD and its quantity reflected the increased severity of inflammation. The combination of microsequencing and mass spectrometry led to the identification of the 13 kDa spot as calgranulin B. Our results indicate that the production of calgranulin B is unregulated in inflammatory, preneoplastic and neoplastic lesions of colonic mucosa.

Adult↗

PubMed1995

Analysis of monoclonal immunoglobulin light chains in urine using two-dimensional electrophoresis.

High-resolution two-dimensional electrophoresis was used to analyze kappa-type monoclonal immunoglobulin light chains in urine of two patients with multiple myeloma. The patients were treated, remission lasted about 2.5 years. In the period of relapse of the disease, acid isoforms of kappa chains, pI < 4,95, which had not been present in the urine of these patients before, were found in urine of both patients. Kappa-type Bence-Jones protein in urine of one patient showed unusual behavior: It failed to precipitate in the range of 40-60 degrees C or on boiling.

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PubMed1995

Electrophoretic analysis of microheterogeneity of paraproteins in a patient with IgD myeloma.

High-resolution two-dimensional polyacrylamide gel electrophoresis (2-DE) was used to analyze more precisely serum and urine specimens of a patient suffering from IgD myeloma associated with renal insufficiency. The application of 2-DE with immobilized pH gradient followed by immunoblotting revealed the presence of acidic monoclonal delta chains, hidden on 2-DE by albumin. This approach also enabled to detect two other forms of delta heavy chains expressing both reduced (45 kDa) and high (110 kDa) mol. weight. The analysis of urine specimen proved the presence of three acidic isoforms of monoclonal lambda light chains together with multiple monoclonal light chain fragments, which strongly suggests amyloidogenicity of these monoclonal light chains.

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