Preparation and quality control of [166Ho]-macroaggregates for radiosynoviorthesis.

(166)Ho is a beta-emitter (E(_max) = 1.84 MeV, T1/2 = 26.7 hours) with a mean penetration in the soft tissue of 2.2 mm and gamma-photons 81keV and 1380keV. In this paper, we present our experiences with preparation of (166)Ho-macroaggregates ((166)Ho-MA) for radiosynoviorhesis. Ho-MA were prepared by reacting the aqueous solution of holmium nitrate pentahydrate with sodium borohydride solution in 0.2 M NaOH. After centrifugation, washing, and drying at 105 degrees C, final fineness of 1-15 microm was attained using a homogenizer working at 70,000 rpm. Neutron activation was carried out in screwed titanium ampules in a nuclear reactor at the neutron flux 10E12-10E14 neutrons.cm-24.s-1. The irradiated samples were allowed to cool 24 hours before further manipulation. (166)Ho-MA were clinically tested on 20 patients suffering from gonarthrosis (15), rheumatoid arthritis (5), psoriatic arthritis (1), and gout arthropathy (1). (166)Ho-MA are currently produced under GMP and production facility allows producing radioactivity for 6 patients within one production run.

Our first clinical experience with radiosynoviorthesis by means of (166)Ho-holmium-boro-macroaggregates.

BACKGROUND: In this paper, we evaluate the therapeutic and adverse effects of the application of 166-holmium-boro-macroaggregates (HMBA) in radiosynovectomy (RSO) of the knees. We assessed the efficacy and safety of (166)Ho-HBMA in a prospective clinical trial in patients suffering from chronic synovitis. MATERIAL AND METHODS: An effective component of radiopharmaceutical (166)Ho-boro-macroaggregates is radionuclide (166)Ho which has both beta-emission and gamma-emission. The physical half-life time of 166 Ho is 26.8 hours. After application of the radiopharmaceutical into a joint cavity, the effect of beta-emission causes radiation necrosis of pathologically changed (inflamed) synovial membrane. From 15th April 2005, we have started RSO of knees by means of new radiopharmaceutical (166)Ho-boro-macroaggregates in patients with gonarthrosis, rheumatoid arthritis, chronic synovitis, psoriatic arthritis, gout arthropathy. Seventeen intra-articular injections were performed in fifteen patients receiving a mean activity of 972 MBq (range: 904-1,057 MBq) (166)Ho-HMBA. The patients were hospitalized for three days. Side effects were evaluated during hospital stay and after 6-8 weeks. Static scintigraphy of knee joints and measurements of blood radioactivity were performed. Therapeutic effects were evaluated after 6-8 weeks. RESULTS: In 2 hours and 2 days after application, we proved, by means of knee and inguinal scintigraphy, only insignificant radiopharmaceutical leakage from the joint cavity to the inguinal lymph nodes in four patients. In treated patients, no serious adverse effects occurred. Nine patients were without complaints; 4 patients had slight knee exsudation and 2 patients had great exsudation. Therapeutic effects after 6-8 weeks were as follows: 2 patients were without pain, 9 with lower pain, 3 with the same pain and 1 patient with increased pain. Joint motion was improved in 7 patients, remained the same in 7 patients and was impaired in 1 patient. Analgesics consumption was lower in 5 patients, the same in 9 patients and greater in 1 patient. Knee exsudation was absent in 2 patients, lower in 4 patients, the same in 6 patients and greater in 3 patients. CONCLUSIONS: We proved only insignificant radiopharmaceutical leakage from the joint cavity to the inguinal lymph nodes. Six patients had early slight or great radiation synovitis. The possible cause could be rather high applicated activity. One can take into consideration its reduction. Therapeutic effects can be precisely evaluated after a longer time interval than was possible for us (6-8 weeks after RSO). (166)Ho-boro-macroaggregates can extend the scale of clinically used radiopharmaceuticals for RSO. This paper is presented in the scope of the first stage of clinical evaluation of synovectomy application of holmium-boro-macroaggregates.

[Dealing with Rhizarthrosis by Means of Arthrodesis with Finger Micro-plate Osteosynthesis.]. / Resení rizartrózy s pouzitím prstové dlahy z miniinstrumentária AO V.

The authors of the article describe possibilities of dealing with thumb CMC joint degenerative changes, concentrating on arthrodesis. They compare individual operations and refer about fixation of arthrodesis with micro plate osteosynthesis which enables early functional treatment. They perform the arthrodesis in patients with radiologically proved arthrosis, in most cases of stage III, who do not respond to conservative therapy. They state both short-term and long-term results in their own group of 18 operations (all of the operated patients state improvement of thumb function and relief of pain following the operation) and analyse reasons for failure of the technique and for the necessity of revision in 3 of them. Key words: carpo-metacarpal I. joint, arthrodesis, osteoarthritis of trapezio-metacarpal joint.

[Ultrasonographic Examination of the Rotator Cuff of the Shoulder: Our Eight-year Experience.]. / Ultrasonografické vysetrení rotátorové manzety ramenního kloubu - nase osmileté zkusenosti.

The authors evaluate their experience with ultrasonographic (USG) examinations of the shoulder they performed in the period between 1992 and 1999. They concentrate mainly on the diagnosis of the lesion of the rotator cuff. They describe methodology of the examination, the used examination positions, instruments and present some pathological findings. They also present possibilities of these diagnostic methods and the indications where the efficiency of its use is limited or zero.They emphasize the necessity of a perfect mastering of the examination technique, a simultaneous clinical examination, x-ray examination and special x-ray projections, if need be. In total they performed 1000 examinations and processed the total number of 762 examinations. Of this number they performed 499 examinations for a suspected rupture of the rotator cuff. They compare the results with per-operative arthroscopic and open revision findings. In this way they processed a group of 78 patients operated on shoulders with preoperative USG. Key words: ultrasonography of shoulder, examination technique, normal and pathological findins.

Genetic analysis of the rat hypodactylous mutation.

Autosomal recessive rat hypodactylous mutation Hd leads in homozygous condition to reductive changes of the digital arch of all feet. There is a variable preaxial reduction of the number of fingers in both sexes. Moreover, homozygous males are sterile. Testes of homozygous Hd/Hd and +/Hd adult rats were examined in the light and electron microscopes. Spatial organization of stages of the spermatogenetic cycle was not confirmed in Hd/Hd testes comparing with +/Hd males. Significant decrease in the number of germ cells in seminiferous tubules of Hd/Hd testes was accompanied with loosening and vacuolization of the seminiferous epithelium. The assignment of the Hd locus to RNO10 excluded the suspected homology between rat and mouse Hd mutations. More precise mapping using microsatellite markers revealed close linkage of the Hd locus with the D1OMit8 marker defining Syb2 gene coding for synaptobrevin 2. A chromosomal segment of RNO10 carrying Hd and Syb genes is being incrossed onto BN and SHR inbred strains in order to examine the modifying influences of different genetic backgrounds.

Recombinant inbred and congenic strains of the rat for genetic analysis of limb morphogenesis.

Recombinant inbred (RI) and congenic strains carrying the polydactyly-luxate syndrome (PLS) provide an experimental model for the analysis of polygenic control of limb development. PLS is determined by a major gene Lx whose phenotypic expression is strongly influenced by the genetic background upon which it operates. The morphometric analysis of the skeleton of front and hind legs has been carried out. The morphotypes of PLS in RI strains exhibit a continuous variability and transgressive variation compared to BN.Lx and SHR.Lx morphotypes, which strongly indicates the polygenic effects on PLS manifestation. Quantitative trait loci (QTL) were searched for through correlation of genetic markers and morphometric traits. The association analysis revealed statistically significant correlations (P < 0.0003) of morphometric traits with two markers on chromosome 4 (Il6 and A2m) associated with the number of front feet and hind feet phalanges, respectively, one marker on chromosome 7 (D7Mit17) associated with the tibia length, and the somatostatin gene on chromosome 11 associated with the number of front feet phalanges. In addition, suggestive associations of morphometric traits with markers on further nine chromosomes have been found (correlation coefficients ranging from 0.5 to 0.6). The verification of all these findings is in progress by means of double congenic strains which, in addition to the Lx gene, carry differential chromosome segments with putative modifiers.

Contribution to the morphometry of limb bud structures in the normodactylous and polydactylous rat. I. Apical ectodermal ridge.

The height of the apical ectodermal ridge on limb buds of the embryo laboratory rat was studied in the polydactyly-luxate syndrome and compared with the controls. The following findings were obtained: (a) On the 14th embryonal day, prior to the development of the anlage of mesenchymal condensates, the AER is higher in polydactylous animals as compared with the controls. (b) On the 15th, 16th and 17th embryonal day the height of the AER in the praeaxial region of the polydactylous limb bud largely predominates over the controls. A comparison of the height of the AER above digital rays and interdigital grooves of polydactylous and normodactylous animals does not thus exhibit any marked differences. This fact is attributed to the existence of more powerful induction processes of the underlying mesenchymal component where rudiments of supernumerary digital rays are formed.

Contribution to the morphometry of limb bud structures in the normodactylous and polydactylous rat. II. Density of filopodia in the subectodermal zone.

In hindlimb buds of normodactylous and polydactylous embryos in the stage of the 16th and 17th embryonal day the mesenchymal region, closely adjoining the site below the AER, was investigated. This space is called the subridge zone and is filled with a large amount of variously formed processes of mesenchymal cells, chiefly with filopodia. With the use of the morphometric point-couting method it was found that in normodactylous limb buds the density of filopodia in the given area of the subridge zone was 2.15% as compared with the 6.48% representation of filopodia in the same zone of polydactylous animals. Numerous filopodia localized right under the AER established connection between the mesenchyma and the intact basal membrane, and their higher density is undoubtedly related to the existence of the maintenance factor.

Patterns of physiological cell death and mitoses in the apical ectodermal ridge in normodactylous and polydactylous rat limb buds. A quantitative evaluation.

Limb buds in normally developing foetal rats and rats with the polydactyly-luxate syndrome at day 14 and 15 of intrauterine growth were studied at the optical and electron microscopic levels. Four sets of limb buds were collected and evaluated: 14th e.d.N., 14th e.d.P., 15th e.d.N., and 15th e.d.P. (e.d. embryonic day, N - normodactylous, P - polydactylous); they were studied for apical ectodermal ridge development with reference to the rates of cell death and mitosis. Both phenomena were evaluated quantitatively. A significant difference was found in the rate of physiological cell death, which was far lower in polydactylous (4.33 and 28.73) as compared with normodactylous individuals (62.40 and 82.11 cell deaths per 300 cells in the ridge on days 14 and 15). The difference found documents a considerable retardation in cell death rate in the polydactylous apical ectodermal ridge epithelium. Physiological cell death is associated with a process called apoptosis, which is characterized by the presence of "apoptotic bodies" in the cytoplasm of epithelial cells and in the intercellular space. The total number of mitoses in the ridge epithelium was relatively low (4.63 - 6.33 per 300 cells) and the differences between norm and polydactyly were not statistically significant.

Ciliary beat frequency of biopsy samples taken from normal persons and patients with various lung diseases.

Ciliary beat frequency from human tracheobronchial mucosa was measured in vitro. Biopsy samples were taken during bronchoscopy and beat frequency measured with a phase contrast microscope and a photosensitive cell. In 20 healthy volunteers the mean ciliary beat frequency in the trachea was 12.5 +/- 2.8 Hz at 37 degrees C with a wide interindividual range (4.9 to 17.4 Hz), but a good intraindividual reproducibility (+/- 7 percent). There was a slight increase in frequency down the tracheobronchial tree and a decrease at lower temperatures. With some exceptions, no disease-specific correlation in beat frequency seems to exist in different bacterial and malignant bronchopulmonary conditions. However the quantity and quality of ciliated cells brushed from the mucosa does appear to be disease-related.

[Method of photometric measurement of ciliary beat frequency from human tracheobronchial epithelium (author's transl)]. / Eine Methode zur Messung der Ziliarfrequenz aus Bürstenbiopsien der menschlichen Bronchialschleimhaut.

A method of photometric measurement of ciliary beat frequency is described. Brush biopsies of human bronchial epithelium were obtained. Twenty healthy persons were investigated, sampling brush biopsies from trachea, main bronchus and segmental bronchus. Ciliary beat frequency was measured over a period of time at 37 degrees C as well as under increasing temperature from 20 degrees C to 40 degrees C. Mean ciliary beat frequency in the trachea is 12.5 Hz +/- 2.8 at 37 degrees C, slightly increasing ciliary beat frequency towards the periphery of the bronchial tree. Ciliary beat frequency found at 20 degrees C is almost exactly half the frequency found at 37 degrees C. When rising the temperature from 30 degrees C to 40 degrees C, ciliary beat frequency increases nearly in a linear line. A well reproducible method is used to measure ciliary beat frequency in vitro under different conditions.

Influence of reproterol on ciliary beat frequency of human bronchial epithelium in vitro.

With a recently developed method it is possible to measure ciliary beat frequency of human bronchial epithelium in vitro. By means of brush biopsy, mucosa is taken from a region of the tracheo-bronchial tree and is then put into nutrient solution. Action of the cilia can be observed under a phase contrast microscope and is transferred by a photocell onto an oscillograph. The study was performed in 15 patients; mucosal samples were taken from both the trachea and the main bronchus. It proved that the beat frequency of cilia obtained from both the trachea and the main bronchus was significantly increased by 7-[3-(beta, 3,5-trihydroxyphenethylamino)-propyl]-theophylline (reproterol, Bronchospasmin), as compared to those samples without reproterol.