Biomedical subjects

A Urban

Publications and source records attributed to A Urban.

At least 19 recordsLinked to original sources

PubMed2003

Optimisation of expression and purification of the recombinant Yol066 (Rib2) protein from Saccharomyces cerevisiae.

Yeast protein Yol066 (encoded by YOL066 ORF, also known as Rib2) possesses two distinct sequence domains: C-terminal deaminase domain and N-terminal part related to RNA:pseudouridine (psi)-synthases. The deaminase domain is implicated in the riboflavine biosynthesis, while the exact function of the RNA:Psi-synthase domain remains obscure. Here we report the optimisation of growth conditions and purification scheme for recombinant His(6)-tagged Yol066 expressed in E. coli BL21(DE3) using pET28 plasmid. Production of soluble Yol066 protein is best at low temperature (18 degrees C) and IPTG concentration (50 micro M) and Yol066 purification was achieved using metal-affinity and ion-exchange chromatography. This optimised protocol yields about 10 mg of highly purified recombinant Yol066 from 3 l of E. coli culture.

Chromatography, Ion Exchange↗

PubMed2003

Comorbidity of parkinsonism and schizophrenia in a patient treated with clozapine.

Clozapine is the least likely anti-psychotic to induce extrapyramidal symptoms (EPS). We present a surprising case of a woman schizophrenic patient treated with clozapine suffering from EPS. Single photon emission computed tomography (SPECT) revealed a low density of presynaptic dopamine transporters in our patient's brain. A comorbid diagnosis of Parkinson's disease in schizophrenia was confirmed in this way. This helped us to find a proper therapeutic strategy for our patient.

Antipsychotic Agents↗

PubMed2001

DsbA and DsbC affect extracellular enzyme formation in Pseudomonas aeruginosa.

DsbA and DsbC proteins involved in the periplasmic formation of disulfide bonds in Pseudomonas aeruginosa were identified and shown to play an important role for the formation of extracellular enzymes. Mutants deficient in either dsbA or dsbC or both genes were constructed, and extracellular elastase, alkaline phosphatase, and lipase activities were determined. The dsbA mutant no longer produced these enzymes, whereas the lipase activity was doubled in the dsbC mutant. Also, extracellar lipase production was severely reduced in a P. aeruginosa dsbA mutant in which an inactive DsbA variant carrying the mutation C34S was expressed. Even when the lipase gene lipA was constitutively expressed in trans in a lipA dsbA double mutant, lipase activity in cell extracts and culture supernatants was still reduced to about 25%. Interestingly, the presence of dithiothreitol in the growth medium completely inhibited the formation of extracellular lipase whereas the addition of dithiothreitol to a cell-free culture supernatant did not affect lipase activity. We conclude that the correct formation of the disulfide bond catalyzed in vivo by DsbA is necessary to stabilize periplasmic lipase. Such a stabilization is the prerequisite for efficient secretion using the type II pathway.

Alkaline Phosphatase↗

PubMed2001

[Photodynamics in gynaecology].

Photodynamic procedures were intended for early detection and therapy of neoplastic diseases. Fluorescence diagnosis is based on the detection of the fluorescence of exo- and endogenous photosensitizers selectively accumulated in pathological tissues. Photodynamic therapy is based on the selective destruction of neoplastic tissue resulting from interaction between photosensitizer, light and oxygen. The main fields of the clinical use of photodynamics are pulmonology, gastroneterology, urology and dermatology. The article presents the theoretical principles of fluorescence diagnosis and photodynamic therapy followed by the review of up-to-date literature concerning the use of photodynamics in diseases of the genital tract in women.

Female↗

PubMed2000

Rapid gene inactivation in Pseudomonas aeruginosa.

A rapid and efficient method to inactivate genes in Pseudomonas aeruginosa has been developed. It is based on pKnockout vectors which carry either a gentamicin or a streptomycin/spectinomycin resistance cassette allowing for selection in P. aeruginosa where these vectors do not replicate. A PCR fragment of the gene of interest carrying 5'- and 3'-truncations is cloned into a pKnockout vector, mobilized into P. aeruginosa, and subsequently integrated into the chromosomal copy of the target gene. The orientation of the fragment determines whether (i) the target gene is disrupted without blocking the transcription of downstream genes or (ii) the insertion exerts a polar effect thereby leading to inactivation of a whole operon.

Conjugation, Genetic↗

PubMed2000

[Eye complications in 458 children with rheumatoid arthritis].

BACKGROUND: Besides arthritis or other systemic manifestations, endogenous inflammation (most commonly uveitis) can be a symptom of rheumatoid arthritis in children. The uveitis of children shows certain differences compared to that in adults. This study will show these differences and their incidences. PATIENTS AND METHODS: Retrospective analysis of the histories of 458 children that were admitted to the pediatric clinic due to rheumatism related diseases. An analysis of the general and pediatric data, laboratory results and therapy were combined with the ophthalmologic data. RESULTS: 45 (9.8%) of the 458 patients suffered from uveitis at least once during the studied time period between 1992 and 1996. Most common complications of the therapy were clouding of the lens up to full cataract (in both eyes in eight out of nine patients). Seven patients underwent cataract operation. Further complications are synechiae, band shaped keratopathy, secondary glaucoma and retinal diseases. CONCLUSION: Since the pediatric uveitis usually does not take an acute course and since children rarely can articulate themselves precisely, a regular ophthalmologic examination of pediatric rheumatology patients is necessary, as well as every uveitis in children should warrant pediatric-rheumatologic diagnostics. The impact of uveitis is much more severe for children when compared to adults, since it not only endangers the vision but the whole development of the child.

Age Factors↗

PubMed2000

[Use of photodynamic methods in diagnosis and therapy of epithelial vulvar diseases].

Therapy of squamous vulvar diseases creates main problems in modern gynecology. Except some useful and tried, but sometimes not optimal, therapeutical methods, medicine still discovers new diagnostic and therapeutical procedures. Authors describe new trends in photodynamic diagnostics and treatment (PDD and PDT) of some vulvar epithelial diseases: condylomata acuminata and squamous cell vulvar cancer.

Aminolevulinic Acid↗

PubMed1999

Palliative radiotherapy for recurrent and metastatic malignant melanoma: prognostic factors for tumor response and long-term outcome: a 20-year experience.

PURPOSE: Radiotherapy is used as a "last resort" for patients with advanced cutaneous malignant melanoma. We have analyzed our 20-year clinical experience with respect to different endpoints and prognostic factors in patients with locally advanced, recurrent, or metastatic malignant melanoma. METHODS: From 1977 to 1995, 2,917 consecutive patients were entered in the melanoma registry of our hospital. Radiotherapy was indicated in 121 patients (56 females, 65 males) for palliative reasons in advanced malignant melanoma stages UICC IIB/III/IV. The histology of the primary lesion was nodular in 51 patients, superficial spreading in 35, acral-lentiginous in 8, and lentigo maligna melanoma in 4 patients. Eleven patients had primary or recurrent lesions which were either not eligible for surgery or had residual disease (R2) after resection of a primary or recurrent lesion (UICC IIB); 57 patients had lymph node (n = 33) or in-transit metastases (n = 24) (UICC III), and 53 had distant organ metastases (7 M1a; 46 M1b) (UICC IV). Time from first diagnosis to on-study radiotherapy averaged 19 (median: 18; range: 3-186) months. In most cases, conventional RT was applied with 2-6 Gy single fractions up to a median total radiation dose of 48 (mean: 45; range: 20-66) Gy. RESULTS: At 3 months follow-up, complete response (CR) was achieved in 7 (64%) and overall response [complete (CR) and partial response (PR)] in all (100%) UICC IIB patients, in 25 (44%) and 44 (77%) of 57 UICC III patients, and in 9 (17%) and 26 (49%) of 53 UICC IV patients. Tumor progression during radiotherapy occurred in 25 (21%) patients. Patients with CR survived longer (median: 40 months) than those without CR (median 10 months) (p < 0.01). At last follow-up (Dec 31, 1996), 26 patients were still alive: 6 (55%) UICC IIB, 17 (30%) UICC III, and 3 (6%) UICC IV patients (p < 0.01). Univariate analysis revealed the following prognostic factors for complete response and long-term survival: UICC stage (p < 0.001), primary location in the head and neck region, total radiation dose above 40 Gy (all p < 0.05), while age, gender, and histology had no impact. In multivariate analysis, UICC stage was the only independent prognostic factor (p < 0.001). CONCLUSION: External beam radiotherapy can provide long-term local control and effective palliation in malignant melanoma UICC stages IIB-IV. The current UICC staging system is an excellent prognostic factor for initial and long-term tumor response in metastatic melanoma. Therefore, prospective randomized trials using external radiotherapy with or without adjuvant therapy for advanced malignant melanoma are justified.

Adult↗

PubMed1999

Distribution and developmental changes in metabotropic glutamate receptor messenger RNA expression in the rat lumbar spinal cord.

Using in situ hybridisation, the regional distribution of primary transcripts and splice variants of all metabotropic glutamate receptor subtypes (mGluR) currently known to be expressed in the spinal cord have been studied in the lumbar enlargement of the rat spinal cord. In adult animals, the messenger RNA of the mGluR subtypes 1, 5, 3, 4 and 7 were differentially expressed. The transcripts of mGluR1 and 5 were most abundant with mGluR5 messenger RNA being concentrated in the superficial dorsal horn. In contrast, the mGluR2 transcript was not detectable with the sensitivity of the method. Secondly, age related changes (postnatal days 1, 7, 12, 21) in the postnatal expression of mGluR1-5 and 7 transcripts have been investigated. mGluR1 and 7 messenger RNA showed a general decrease in spinal expression from postnatal day 1 to day 21. Quantitative densitometry showed high mGluR3 and 5 messenger RNA levels especially in the superficial dorsal horn at birth, however these levels decreased with age. In addition to changes in density, the regional distribution of mGluR3 messenger RNA was altered with postnatal development. Up to postnatal day 12, mGluR3 messenger RNA expression was almost exclusively restricted to the spinal grey matter, but with postnatal day 21 a strong additional expression in the white matter occurred. Distribution of mGluR4 messenger RNA showed little change in the dorsal horn, however motoneuronal expression emerged during development. These changes may suggest different roles for mGluRs in the maturation of spinal transmission of the rat nervous system.

Aging↗

PubMed1999

[Locally recurrent and metastatic malignant melanoma. Long-term results and prognostic factors after percutaneous radiotherapy].

PURPOSE: Radiotherapy (RT) is used as last resort for patients with advanced cutaneous malignant melanoma (MM). Herein our 20-year clinical experience is presented analyzing different endpoints and prognostic factors in patients with locally advanced, recurrent or metastatic MM. PATIENTS AND METHODS: From 1977 to 1995, 2,917 consecutive patients were entered in the MM registry of our university hospital. RT was indicated in 121 patients (56 females, 65 males) for palliation in locally advanced recurrent and metastatic MM stages UICC IIB to IV. At the time of RT initiation, 11 patients had primary or recurrent lesions which were either not eligible for surgery or had residual disease (R2) after resection of a primary or recurrent MM lesion (UICC IIB); 57 patients had lymph node (n = 33) or in-transit metastases (n = 24) (UICC III), and 53 had distant organ metastases (7 M1a, 46 M1b) (UICC IV). The time from first diagnosis to on-study RT averaged overall 19 months (median: 18; range: 3 to 186 months). In 77 patients conventional RT and in 44 patients hypofractionted RT was applied with 2 to 6 Gy fractions up to a mean total RT dose of 45 (median: 48; range: 20 to 66) Gy. RESULTS: At 3 months follow-up, complete response (CR) was achieved in 7 (64%), overall response (CR + PR) in all (100%) UICC IIB patients, in 25 (44%) and 44 (77%) of 57 UICC III patients, and in 9 (17%) and 26 (49%) of 53 UICC IV patients. Tumor progression during RT occurred in 25 (21%) patients. Patients with CR survived longer (median: 40 months) than those without CR (median 10 months) (p < 0.01). At the time of evaluation and last FU (December 31, 1996), 26 patients were still alive: 6 (55%) stage UICC IIB, 17 (30%) stage UICC III, and 3 (6%) stage UICC IV patients (p < 0.01). Univariate analysis revealed following prognostic factors for CR and long-term survival: UICC stage (p < 0.001), primary location in the head and neck, total RT dose > 40 Gy (all p < 0.05), while age, gender and primary histological subtype had no impact. In multivariate analysis, UICC stage was the only independent favorable prognostic factor for achievement of CR and long-term survival (p < 0.001). CONCLUSION: External RT provides effective palliation in advanced UICC stages. The UICC staging system is a good predictor of initial and long-term tumor response in metastatic MM. Prospective randomized trials using RT with or without adjuvant therapy for advanced MM are justified.

Adult↗

PubMed1999

[Long term results following radiation therapy of locally recurrent and metastatic malignant melanoma].

The 20-year radiotherapy (RT) experience in patients with locally advanced, recurrent or metastatic malignant melanoma (MM) is analyzed with respect to different endpoints and prognostic factors. From 1977 to 1995, 2917 consecutive patients were entered in our MM registry. RT was indicated in 121 patients (56 females, 65 males) for palliation in advanced MM stages. The histology of the primary lesion was nodular in 51, superficial spreading in 35, acral-lentiginous in 8 and lentigo maligna in 4 patients); 22 were missing or could not be reclassified. Eleven patients had primary or recurrent lesions which were ineligible for surgery or had residual disease (R2) after resection of a primary or recurrent lesion (UICC IIB); 57 patients had lymph node (33) or in-transit metastases (24) (UICC III), 53 had distant organ metastases (7 M1a; 46 M1b) (UICC IV). Time from first diagnosis to on-study RT averaged 19 (median: 18; range: 3-186) months. In most cases conventional RT was applied (2-6 Gy single fractions) up to a mean total RT dose of 45 (median: 48; range: 20-66) Gy. At 3 months follow-up (FU), complete response (CR) was achieved in 7 (64%) and overall response (CR+PR) in all (100%) UICC IIB patients, in 25 (44%)/44 (77%) of 57 UICC III patients, and in 9 (17%)/26 (49%) of 53 UICC IV patients. Progression during RT occurred in 25 (21%) patients. Patients with CR survived longer (median: 40 months) than those without CR (median: 10 months) (p<0. 01). At last FU, 26 patients were alive: 6 (55%) UICC IIB, 17 (30%) UICC III, and 3 (6%) UICC IV patients (p<0.01). In univariate analysis following favorable prognostic factors for CR and long-term survival were identified: low UICC stage (p<0.001), primary site head and neck and total dose >40 Gy (all p<0).

Adult↗

PubMed1999

[Results of six-years-term perinatal care of diabetic woman in Silesia district].

Authors had retrospectively analyzed basic exponents of perinatal care of 256 pregnant diabetic woman in years 1993-1998 hospitalized and delivered in Clinic of Obstetrics and Gynaecology in the town of Bytom. Our results were very similar to the other of Referent Centres Of Technical Methods Bank Used in Diagnostic and Therapy of Diabetes: cesarean sections percentage 51.56%, 4.7% of undelivered pregnancies, 2.34% of new-born delivered with multiple congenital defaults, 17.2% of macrosomies and 39.8% pre-term deliveries were noted. Disadvantage perinatal effects in diabetic women might be reduced by very early diagnosis of diabetic and intensive biophysical and biochemical fetal care, which is strongly suggests by the authors.

Catchment Area, Health↗

PubMed1999

[Analysis of indications for cesarean section in pregnancy complicated by diabetes mellitus].

The authors analysed a way of pregnancy ending and indications to cesarean section in group 256 diabetic pregnant women. We estimated that percentage for cesarean sections was 51.56%. We noticed downward tendency of elective indications to cesarean section (the most common was gestosis) and upward tendency of intrapartum indications (the most common was fetal distress syndrome). The percentage of cesarean sections in diabetic pregnant women group in our study is similar to results from the other centres.

Cesarean Section↗

PubMed1998

[Results of five-year term perinatal care of a diabetic woman].

Authors had retrospectively analysed basic exponents of perinatal care of 199 pregnant diabetic woman in years 1993-1997 hospitalised and delivered in The First Clinic of Obstetrics and Gynaecology in the town of Bytom. Our results were very similar to the other Referent Centres Of Technical Methods Bank Used in Diagnostic and Therapy of Diabetes: average gestation age 38 +/- 1 hbd, caesars sections percentage 53.4%, 5.5% of undelivered pregnancies, 0.5% of new-born delivered with multiple congenital defaults, 16.1% of macrosomies and 28.1% pre-term deliveries were noted. Disadvantage perinatal effects in diabetic women might be reduced very early diagnosis of diabetic and intensive biophysical and biochemical fatal care, which is strongly suggests by the authors.

Female↗

PubMed1997

A rapid and efficient method for site-directed mutagenesis using one-step overlap extension PCR.

A rapid method is described to efficiently perform site-directed mutagenesis based on overlap extension polymerase chain reaction (OE-PCR). Two template DNA molecules in different orientations relative to only one universal primer were amplified in parallel. By choosing a high dilution of mutagenic primers it was possible to run an overlap extension PCR in only one reaction without purification of intermediate products. This method which we have named one-step overlap extension PCR (OOE-PCR) can in principle be applied to every DNA fragment which can be cloned into a multiple cloning site of any common cloning vector.

Animals↗

PubMed1997

Bronchiolitis obliterans organizing pneumonia associated with acute Mycoplasma pneumoniae infection.

A patient with bronchiolitis obliterans organizing pneumonia (BOOP) associated with acute Mycoplasma pneumoniae infection is described. Although mycoplasmas have occasionally been associated with bronchiolitis obliterans, to the best of our knowledge, this is the first well-documented case of BOOP associated with M. pneumoniae infection. The diagnosis of BOOP was made by open-lung biopsy after fiberoptic transbronchial biopsy proved nondiagnostic. Corticosteroid therapy was administered and a dramatic improvement was observed; the patient remained without complaints during a 3-year follow-up period.

Acute Disease↗

PubMed1997

[Evaluation of the dynamics of in-group strain during long-term isolation].

Evaluation of the dynamics of in-group strain was conducted with conjunction with the 90-day modelling of space flight (ECOPSY-95). Baseline were the data of sociometric testing and analysis of video records of movements and behavior of the main and visiting crews (three members in each). Global assessment of the in-group strain was made on integration of experimental data using the Czech procedure of interdisciplinary approach to description of a social system, and the theory of fuzzy aggregate. The procedure allows to visualize interpersonal relations as social maps. Reaction to the psychoemotional strain stemmed from long stay in isolation was manifested by gradual build-up of interpersonal strain, redistribution of the functional/role structure of the crew, destabilization of in-group interactions and its split.

Adult↗

PubMed1996

Fine-needle aspiration biopsy of metaplastic carcinoma of the breast: report of a case with abundant myxoid ground substance.

Metaplastic carcinoma (MC) is an uncommon neoplasm of the breast. There are several variants of MC depending on the dominant histologic pattern. The components include over infiltrating ductal carcinoma, extensive squamous differentiation and spindle cell proliferation with or without chondroid or asseous heterologous elements. In FNA smears, only 57% of cases show both ductal carcinoma and metaplastic component. Thus, in almost one half of the cases, the diagnosis is not possible by FNA. Often it is difficult to define the epithelial or sarcomatous character of malignant cells. We describe a case of metaplastic carcinoma of the breast studied by fine-needle aspiration cytology in which myxoid ground substance was the dominant feature in the cytology smears. The rest of the material was composed of scanty isolated atypical cells with large and irregular nuclei. It is important to bear in mind the diagnosis of MC and make a careful search for atypical cells when the cytological smears are mainly composed of myxoid ground substance.

Biopsy, Needle↗