[Bone healing capacity in patients undergoing total hip arthroplasty]. / Schopnost kostní reparace u pacientu podstupujících implantaci TEP kycelního kloubu.

PURPOSE OF THE STUDY: To identify some characteristics of bone repair capacity in elderly patients who undergo total hip arthroplasty, which requires good healing ability of bone for implant osteointegration and bone defect repair, particularly if revision arthroplasty is necessary. MATERIAL AND METHODS: In a group of 27 patients (mean age, 70 ± 7 years; range, 60 to 81 years) a coincidence of osteoarthritis and osteopenia/ osteoporosis was assessed, and mesenchymal stem cells (MSC) were isolated and their numbers, viability and proliferative capacity were evaluated. The MSC populations were examined for their behaviour on bone tissue scaffolds used in orthopaedic surgery for treatment of bone lesions. Each patient underwent bone densitometry examination before total hip arthroplasty. Bone marrow was harvested intra-operatively from the trochanteric region of the femur. From a portion of bone marrow, MSCs were isolated and cultured, and a mononuclear cell concentrate was obtained. Either whole bone marrow or a mononuclear cell concentrate was applied to selected matrices (allograft, demineralised bone matrix, porous beta-tricalcium phosphate (-TCP), pressed hydroxyapatite or calcium sulphate). The production of new collagen and extracellular mineralized matrix were first assessed in expansion medium and, when the production was low, differentiation medium was used. RESULTS: A coincidence of osteoarthritis and osteopenia/osteoporosis was found in 50% of the patients. All were women with a low body mass index and had been post-menopausal for an average of 23 years. The isolated MSCs contained a high percentage of viable cells (mean, 95%). The mesenchymal cells of patients with osteopenia, as compared with those having normal bone density, showed markedly lower numbers of fibroblastic colony forming units (CFU-F) per ml and had a lower proliferative capacity because the population doubling time during the first four passages was much longer. Of the scaffolds tested, allografts showed the most marked collagen and extracellular mineralized matrix production in expansion medium with either whole bone marrow or a monocyte concentrate; porous -TCP was the best of bone graft substitutes in collagen and extracellular mineralized matrix production by both whole bone marrow and a monocyte concentrate, but this was only in differential medium. DISCUSSION: The coincidence of ostearthritis with osteopenia/osteoporosis was found in a higher number of our patients than is reported in the literature. Also, a lower MSC proliferative capacity and a low number of CFU-F/ml in the patients with low bone density were interesting findings. Better bone regeneration would generally be achieved with higher MSC numbers and the use of growth factors for stimulation of osteoinduction and angiogenesis. Bone marrow harvesting for MSC isolation, cultivation and subsequent transplantation is currently feasible only in an experiment. A bone marrow aspirate can be applied, but it may not provide a sufficient number of MSCs. In addition to autologous bone grafts, the best collagen production was on allografts. In bone graft substitutes, the porous structure played an important role because on a non-porous material (calcium sulphate) the formation of collagen was very low. There was no difference in collagen and extracellular mineralized matrix production between whole bone marrow and monocyte concentrates. CONCLUSIONS: Elderly patients have reduced bone healing capacity also because of osteopenia/osteoporosis that occurs more often than it is generally diagnosed, including its coincidence with osteoarthritis. The mesenchymal stem cells isolated from osteopenic bone give a lower number of CFU-F/ml and have a lower proliferative capacity. Of the matrices for new bone formation, allografts showed the best results because collagen was produced already in expansion medium. Of the graft substitutes, porous -TCP was the best, but with collagen production in differential medium. The use of bone marrow aspirate is currently a method of choice in order to increase MSC numbers at the site of bone healing. The use of growth factors is an expensive treatment. To achieve the goal of reliable promotion of osteogenesis with cultured MSC transplantation and use of composite materials with pro-osteogenic and pro-angiogenic factors will still require many experimental and clinical studies.

[Allografts in the vascular surgery]. / Alografty v cévní chirurgii.

The use of allografts represents one of the therapeutic options in the treatment of vascular prosthetic infection. Close cooperation with a transplant center is unconditional for successful graft harvest and storage. Although the transplantation programme in the Czech Republic is handled in several centers across the country the use of vascular allografts is still exceptional. During multiorgan harvest procedures arterial or venous grafts have been removed and stored in antibiotic solution at 4 degrees C till implantation. Cardioplegic and short-term organ preserving solution Custodiol was used in our cohort. At the Department of Vascular Surgery of the Na Homolce Hospital in Prague fresh arterial allografts were used in 14 patients in the years 2001-2004. In all, absence of usable autologous graft was the common denominator. In ten cases previous synthetic vascular grafts were infected. Of these, critical limb ischaemia and imminent amputation due to the poor outflow tract occurred in three patients. Once, false femoral artery aneurysm infected with methicilin-resistant Staphylococcus aureus (MRSA) led to several hemorrhagic episodes. There was no early mortality but one early occlusion with subsequent major amputation. In 11 patients (79%) the procedure was uneventful at the short-term follow-up. All patients were treated with cyclosporine.

Changes in cholinesterase activities after daunorubicin administration to rabbits.

1. Acetyl- and butyrylcholinesterase (AChE, BuChE) activities were studied in rabbits with experimentally induced daunorubicin cardiomyopathy. 2. A significant decrease of the plasma BuChE in the daunorubicin group was observed. 3. In the daunorubicin group, AChE activity in the heart was significantly decreased only in the interventricular septum. 4. BuChE activity was significantly decreased in the cardiac septum and ventricles and in the liver following daunorubicin treatment. 5. Changes in cholinesterase activities are probably caused by an effect of daunorubicin oon protein synthesis during the development of certain types of cardiomyopathy.

Increasing quality of anatomical education by introduction of digital imaging into documentation of topographic dissections.

Knowledge of topographic anatomy is essential prerequisite for any physician to be successful in professional career. That is why we pay special attention to topographic dissections completing course of normal human anatomy at our department. Dissection procedure is subjected to relatively rigid rules demanding removal of some superficial structures during preparation of deeper layers. Thus some important structures get lost for further learning and also for final evaluation of students knowledge and manual skills. Therefore we decided to introduce digital imaging into the recording of course of preparation, the topographic relationships and unique variations. Digital acquisition of pictures through still digital camera makes possible to bypass the wet photographic process and provides instant graphic inputs for further computer processing. The image files can be further edited, printed, archived on various media and then may be used for instructional, test, evaluative, and publication purposes. Moreover, own original pictorial material is feasible to be presented at anatomy lectures. Practical results of this new approach, besides higher attractiveness and motivation impact, allow us to conclude that systematic digital acquisition of dissection pictures combined with computer processing fairly increases quality of anatomical education.

Influencing of spatial memory in rats by DSP-4 and mescaline.

Behavioural effects of two experimental neurotoxins, mescaline and DSP-4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine), on retention of spatial orientation were studied in the T-maze. The stereotaxic administration of both neurotoxins into the selected brain structures was chosen to reveal this effect. The intensity and time course of the neurotoxic effect were dependent on the brain area administered. Nevertheless, the lengthening of the latencies in reaching the goal was generally more marked after mescaline in comparison with DSP-4.

Activity of alkaline phosphatase in the major salivary glands of mice at various ages of postnatal life, and during pregnancy and lactation.

Activity of alkaline phosphatase in the major salivary glands of male and female mice at various ages of postnatal life, and in females during lactation was studied histochemically. Enzyme activity was not detected on the day of birth, but was found in the terminal tubules of all major salivary glands during the first postnatal week. Alkaline phosphatase activity was increasing gradually with age and a definitive enzymatic pattern was observed by the age of 6 weeks. No difference in enzyme activity was found among the major salivary glands of young adult and old animals. The parenchyma of fully differentiated submandibular glands showed clear sexually dimorphic patterns of alkaline phosphatase activity. During pregnancy, a significant increase of alkaline phosphatase activity was detected in submandibular gland. From gestation day 15 to the end of pregnancy, enzymic pattern of granular convoluted tubules of pregnant females was the same as in the adult males. Histochemical masculinization of the submandibular gland during pregnancy suggests that besides androgens also progesterone exerts masculinization of the murine submandibular salivary gland.

Toxicities of O-alkyl S-(2-dialkylaminoethyl) methyl phosphonothiolates (V-compounds).

Toxicities expressed as LD50 values of O-alkyl S-(2-dialkylaminoethyl) methyl phosphonothiolates in different species and different routes of administration were determined. Rats were more sensitive to these compounds than mice, rabbits and guinea-pigs were more sensitive than mice and rats. The most sensitive to these compounds were dogs. LD50 values varied in a wide range, however, a decrease of toxicity was observed in the following rank: i.v., i.m., s.c., i.p., p.o. and p.c., respectively. Depending on the route of administration, LD50 values varied from 5 g/kg (i.m., dogs) through tens of microgram/kg for the most part of compounds and routes of administration to hundreds, rarely thousands of g/kg (some chemicals, p.c. administration).

[Development of dipeptidyl peptidase IV (DPP IV) activity in postnatal differentiation of the submandibular gland in mice]. / Vývoj aktivity dipeptidyl peptidázy IV (DPP IV) v prubehu postnatální diferenciace submandibulární zlázy mysi.

The localization of a serine exopeptidase--DPP IV--in the male and female submandibular gland of the mouse during gland postnatal development was studied histochemically in the light microscopic level. The present results suggest that localization of DPP IV is closely related to the postnatal differentiation and maturation of acini and male granular convoluted tubules. Remarkable sex differences of DPP IV activity were detected in the submandibular gland of pubescent and adult animals. The possible physiological role of DPP IV in major salivary glands is also discussed in the paper.

[Histochemical activity of dipeptidyl peptidase IV (DPP IV) in intracerebral transplantation of the submandibular gland in mice]. / Histochemická aktivita dipeptidyl peptidázy IV (DPP IV) v intracerebrálních transplantátech submandibulární zlázy mysi.

Histochemical activity of DPP IV in the intracerebral homotransplants of submandibular gland of the newborn mouse was investigated in the course of a 5-month period after transplantation. Eight weeks after grafting, the enzyme pattern in the parenchyma of transplants was already comparable with the enzyme pattern of a fully mature mouse submandibular gland in situ. At this time, DPP IV was active in the apicolateral cell membranes of numerous acini of the gland transplants. Enzyme activity in apical cell cytoplasm of granular convoluted tubules was found only in transplants located in the brain of male recipients.

[Findings of massive pneumatization of the middle nasal turbinate in a collection of skulls from the 13th-18th centuries]. / Nálezy masivní pneumatizace strední skorepy nosní v souboru lebek z 13.-18. století.

Ethmoid cells display a considerable variability due to their extensive development. Pneumatization of middle turbinate intermediary one or several ethmoid cells is referred to "concha bullosa", which belongs to the frequent and clinically significant variations of bony nasal cavity. The collection of 309 (176 male and 133 female) adult dry skulls pertaining to the "Broumov Ossuary" (13th-18th centuries) was examined for the presence of concha bullosa. The appearance of the middle turbinate was evaluated by inspection. Only noticeably enlarged middle turbinates were rated as positive finding of concha bullosa. All positive findings of concha bullosa were also verified by their probing; coronal CT examination of the nasal cavity and paranasal sinuses was performed in seven of them. Concha bullosa was diagnosed in 160 skulls (51.77%).

[An interesting paleopathologic finding of healed multiple war injuries of a skull]. / Zajímavý paleopatologický nález vícecetného zhojeného válecného poranení lebky.

The presentation describes an interesting finding of healed multiple war cutting wounds in adult male skull. The examined dry skull belong to the extensive osteologic collection which comes from the abolished "Broumov Ossuary". The set encompasses skeletal remains of the population from the 13th to 18th centuries.

Cryopreserved semilunar heart valve allografts: leaflet surface damage in scanning electron microscopy.

OBJECTIVE: Allograft heart valves (AHV), biological valves of human origin, offer potential advantages over conventional xenografts in terms of superior hemodynamics and, perhaps, better durability. The most important factors for long-term AHV clinical performance are the processing and cryopreservation methods. The aim of this study was to evaluate the impact of current processing protocol on valve tissue morphology, mainly to address the effect of successive processing steps on the leaflet surface structure. For the detection of fine changes in endothelial covering and underlying layers, our own modification of the scanning electron microscopy (SEM) technique was utilized. MATERIAL AND METHODS: The study was based on an investigation of 20 AHV (40 specimens). Fourteen valves came from heart-beating donors (multiorgan harvesting) when the heart could not be transplanted for any reason (donor criteria, availability of recipient and/or logistics). Six were obtained at the time of routine postmortems--non heart-beating donors (NHBD). All specimens were initially fixed in Baker's solution. Tissue samples were dissected, dried with hexamethyldisilazane (HMDS), gold-coated, studied and photographed by SEM (Tesla BS 301). In order to define the integrity of the endothelium, subendothelial layers and the quality of the surface under SEM, a special six-level score system was introduced: 1-intact endothelium, 2-confluent endothelium with structural inhomogeneity, 3-disruption of intercellular contacts, 4-separation of endothelial cells, 5-complete loss of endothelium, 6-damage of subendothelial layers). AHV samples were divided into 4 groups for comparison. One aortic AHV "fresh" control sample obtained from a heart-beating donor was evaluated without any processing and was compared with (i) tissue from AHV obtained from NHBD with warm ischemia of 12 and 48 hours, (ii) samples stored at +4 degrees C in saline for 24 h, (iii) antibiotic-treated tissue for 24 h at 37 degrees C and finally with (iv) cryopreserved valves stored in liquid nitrogen (-196 degrees C) for 6-38 months. RESULTS: Our alternative for drying samples by the HMDS method proved to be suitable for thin membranes of human semilunar valves. We were able to detect early changes in the endothelium after harvesting and denudation of the endothelial covering during preservation with and without freezing. The surface of the AHV samples revealed the typical features and score system determined endothelial cell damage. Control "fresh" sample: score 2, (i) NHBD samples with warm ischemia of 12 h: score 3-4, with warm ischemia of 48 h: score 4-5, (ii) samples stored at +4 degrees C in saline for 48 h: score 5-6, (iii) antibiotic-treated tissue for 24 h at 37 degrees C: score 5, (iv) cryopreserved valves stored in liquid nitrogen for 6-38 months: score 5-6. CONCLUSION: SEM (using HMDS drying) together with other methods may be helpful for the morphological control of processing, cryopreservation and liquid nitrogen storage of AHV. Severe AHV leaflet endothelial destruction was proven on AHV grafts. These changes arose already in the initial steps of tissue processing, just after the donor heart harvesting and then at the time of antibiotic valve graft treatment. These results are considered as the starting point for the development of a better preservation protocol.

Allograft semilunar cardiac valves processing and cryopreservation - morphology in scanning electron microscope.

OBJECTIVE: The most important factors of long term clinical performance of biological heart valve prostheses are methods of processing and cryopreservation. That is why we decided to evaluate the impact of current Allograft Heart Valves (AHV) Bank protocol on valve tissue morphology. Scanning electron microscope (SEM) is a valuable tool for investigation of biological surfaces. In case of cardiac valves it is especially suitable for detection of fine changes in endothelial covering and underlying layers. MATERIAL AND METHODS: "Fresh" aortic and pulmonary AHV samples, harvested from "heart-beating" cadaveric donors, were compared with (1) tissue from AHV obtained from non heart-beating donors, (2) samples stored in 4 degrees C saline for 24 h, (3) antibiotic treated tissue for 24 h at 37 degrees C and finally (4) cryopreserved valves, stored in liquid nitrogen (-196 degrees C) for 6-38 months. All samples were dissected, dried with hexamethyldisilazane (HMDS), gold coated, studied and photographed by SEM (Tesla BS 301). RESULTS: Our alternative method of drying samples by the HMDS method proved to be suitable for thin membranes of human semilunar valves. We were able to detect early changes in the endothelium after harvesting, and denudation of the endothelial covering during preservation with and without freezing. CONCLUSION: SEM (using HMDS drying) along with other methods may be helpful for the morphological control of processing, cryopreservation and liquid nitrogen storage of AHV. According to the current findings we have to avoid washing of AHV in saline after harvesting.

Role of interaction of the tanycytes of the third ventricle of the brain with the vascular bed endothelium in enzymatic activation of the parkinsonism-inducing proneurotoxin MPTP in rat brain.

The authors describe the localization of monoaminooxidases oxidizing the protoxin MPTP to the active neurotoxic MPP+ ion, which induces parkinsonism by destroying the dopaminergic neurons of the nigrostriatum. Apart from some types of magnocellular neurons of the hypothalamus, enzymatic activity is localized in the endothelium of segments of the circumventricular vascular bed communicating with the enzymatically equally positive processes of the tanycytes of the third brain ventricle. The findings are briefly discussed from the aspect of the pathogenesis of experimental parkinsonism.

[Scanning electron microscopy of corrosion preparations of the vascular beds of organs in laboratory rats]. / Rastrovací elektronová mikroskopie korozívních preparátu cévního reciste orgánu laboratorního potkana.

A corrosion casting method of the vascular bed of rat's organs using injection of methyl metacrylate resin was described. The casts were photographed in scanning electron microscope Tesla BS 300. Microphotographs of vascular bed casts of selected organs demonstrate the contributions and possibilities of scanning electron microscopy for the investigation of three-dimensional architecture and topographical relations of blood microcirculation in organs.

Localization of 2-glucuronidase in the marginal sinus of rat spleen.

The study demonstrated lysosomal 2-glucuronidase activity localized in the marginal sinus epithelium of rat spleen. Immunologically, the marginal sinus is a key structure of the spleen and its enzymatic apparatus differs from that of the cells of the red pulp venous sinuses. The possible physiological significance of this finding is briefly discussed.

Changes in microvascularization of some rat organs following soman poisoning.

Vascularization of some organs of rat following soman (O-pinacolyl methylphosphonofluoridate) intoxication was studied using scanning electron microscopy. Corrosion casts were prepared with commercially available methyl metacrylate monomer which was partly polymerized, supplemented with catalyst and accelerator and injected into the prewashed and fixed vascular bed. The obtained corrosion casts were sectioned and trimmed using a stereoscopic light microscope. Microvascular casts of normal organs', bed pattern were studied under scanning electron microscope and compared with vascularization patterns after soman poisoning. Changes in the vascular bed architecture of kidneys, brain, adrenal glands and thymus were described.

Changes of some biochemical and hematological parameters following administration of daunorubicin in rabbits.

The effects of the repeated i.v. administration of daunorubicin (50 mg/m2, once weekly, max. 9 weeks) were investigated in rabbits in vivo to analyze biochemical and hematological changes. Noninvasive polygraphic records were used to evaluate the function of the heart. The administration of daunorubicin induced changes especially in levels of protein (decrease in total protein and albumin) and of some ions (decrease in calcium, magnesium and phosphorus) as well as in hematological parameters (decrease in erythrocytes, leukocytes and thrombocytes). The results obtained correlate with data on mechanisms of daunorubicin toxicity.

Autologous vein stent-graft: feasibility study.

PURPOSE: To evaluate expandable stents healed into vein wall as autologous vein stent-grafts for endoluminal grafting. MATERIALS AND METHODS: Balloon expandable stents were placed into external jugular veins of eight dogs. Stent and vein patency was followed by ultrasonography. Five weeks after stent placement, jugular veins with endothelialized stent were harvested. The autologous vein stent-grafts were divided into two groups. In group A, autologous vein stent-grafts (n = 3) were placed immediately into Baker solution for microscopic examination. In group B, autologous vein stent-grafts (n = 3) underwent mechanical manipulation; they were compressed, mounted on angioplasty balloon, pushed through a 9-F sheath and dilated. The autologous vein stent-graft endothelialization and changes after mechanical manipulation were evaluated by light and electron microscopy. RESULTS: Stent placement was successful in seven dogs. One stent migrated into the pulmonary artery. One well placed stent was damaged by compression dressing and thrombosed. At 5 weeks, gross and microscopic examinations revealed the autologous vein stent-grafts were fully covered by a 0.115- +/- 0.036-mm-thick neointimal layer. Small wall thrombus was observed in one autologous vein stent-graft. Repeated manipulations did not result in any intimal damage or stent loosening in the autologous vein stent-grafts. CONCLUSION: Expandable stents healed into a vein have potential to be used as autologous vein stent-grafts for endoluminal grafting without risk of disruption during percutaneous transcatheter introduction.