Occurrence of two histopathologically different malignancies.

BACKGROUND: Synchronous primary malignancies are defined as two or more malignancies without finding a subordinate relationship in different organs of the patient. Reports of patients with primary malignancies with hepatocellular carcinoma are rare. CASE: We describe the course of the disease in a 71-year-old patient with a synchronous occurrence of two histopathologically different malignancies. CONCLUSION: Consistent screening can detect individual tumors early. Understanding the clinicopathological properties and prognostic factors in synchronous malignancies is important to facilitate the treatment.

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.

[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.

[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.

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.

[Preservation of venous grafts]. / Príspevek ke konzervaci zilních stepu.

The objective of the work was to find an optimal preservation medium for short-term preservation of venous grafts which could be subsequently used to line metal stents. The external jugular vein of dogs (n = 15) was removed surgically, divided into portions and immersed into preservation media. For hypothermic preservation (+4 degrees C) solutions of Optisol (Chiron, USA), University of Wisconsin (Baxter, USA), Eurocollins (Fresenius, GFR) and saline (Bieffe Medital, Italy) were used. For normothermic preservation (+37 degrees C) in an atmosphere with 5% CO2 Dulbecc's medium for tissue cultures (Sigma, USA) was used. During hypothermic preservation the specimens were kept for 24 hours, 3 and 7 days, during normothermic preservation in Dulbecc s medium also for 24 hours, 3 and 7 days. The specimens were evaluated by light microscopy and raster electron microscopy. The results revealed that minimal changes on the endothelia of venous grafts occurred during normothermic preservation in Dulbecc's medium where after 7 days the endothelium did not become detached and the vitality of cells did not change. During hypothermic preservation the solution of Wisconsin University proved most suitable. By addition of 5% human albumin to this solution it proved possible moreover to reduce pyknosis of the endothelial cell nuclei. The specimens kept in saline displayed deformities of the nuclei, oedema and loss of endothelia incl. incipient oedema of the cellular wall already after 24 hours of hypothermic preservation. The authors consider the use of this solution unsuitable for preservation.

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.

Pathomorphological changes in microcirculation of pancreas during experimental acute pancreatitis.

BACKGROUND/AIMS: Severe acute pancreatitis is prevalent in Eastern Bohemia (a part of the Czech Republic) and remains a very difficult problem to manage. Recent studies in treatment there are quite frequent but a direct view into the pancreas during its inflammatory process is very rare. Only information about a normal pancreatic microvascular bed appears to be available. This study was designed to explore pathomorphological changes in pancreatic microcirculation at the start and during the development of acute pancreatitis. METHODOLOGY: A group of 50 laboratory white rats was studied. The acute pancreatitis was induced by the modified method of Siech et al. The method of clamping of biliopancreatic duct and stimulation of external secretory tissue by a cholecystokinin and secretin and oral (orogastric tube) ethanol administration was performed. The pancreatic microvascular patterns were observed by using histochemical and corrosion casts methods. RESULTS: The study of the corrosion casts of pancreatic microcirculation in the scanning electron microscope and histochemical studies demonstrated the visible reduction of the pancreatic microvascular bed 18 hours after induction of acute pancreatitis. The microvascular bed is not fully destroyed until 48 hours of acute pancreatitis. CONCLUSIONS: The model of acute pancreatitis using postoperative application of ethanol to the digestive tract after stimulation of pancreas by cholecystokinine and secretin in the rats seems to be real and useful. The study of the corrosion casts of microcirculation in the scanning electron microscope and histochemical studies demonstrated the visible reduction of the pancreatic microvascular bed 18 hours after induction of acute pancreatitis. The microvascular bed is not fully destroyed until 48 hours of running acute pancreatitis, as some "islets" of the vital tissue still have undestroyed microvessels at this time. Despite the above-mentioned serious changes, restricted pancreatic microcirculation enables blood and medicament distribution to the still intact pancreatic tissue.

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.

[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.

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.

Histochemical localization of succinate dehydrogenase in the mouse submandibular gland during postnatal development.

The activity of succinate dehydrogenase (SDH) was examined histochemically in the parenchyma of murine submandibular gland (SMG) during postnatal development on light-microscopical level. In this study, SDH appears as a good marker of differentiating and mature striated ducts of both sexes and convoluted granular tubules of males. Enzyme activity in glandular parenchyma increased from the 1st day to the 7th week of postnatal life. Definitive enzymatic pattern was seen in the gland aged 7 weeks. Histochemical differences between male and female SMG of mouse were noted.

Histochemical localization of succinate dehydrogenase in intracerebral mouse submandibular gland transplants.

Histochemical localization of succinate dehydrogenase in developing intracerebral auto- and homotransplants of the mouse submandibular gland was investigated in the course of a 5-month period after transplantation. Eight weeks after grafting, the enzyme pattern in the parenchyma of homotransplants was comparable with the histochemical picture of a fully mature submandibular gland in situ. At this time, numerous acini showed a weak activity, very numerous striated ducts a strong activity, and less frequent developing convoluted granular tubules slightly weaker activity, than in the striated ducts. Beginning histochemical differentiation of convoluted granular tubules was noted only in homotransplants, located in the brain of male recipients. On the other hand, only a weak activity in the cytoplasm of non-differentiated duct--like structures of some autotransplants was seen. Homotransplants of non-differentiated submandibular gland of newborn donors were found to be a more suitable transplantation object capable of postnatal development of gland parenchyma than autografts of fully differentiated gland of adult animals that did not enter the cytodifferentiation stage.

[Normal and experimental microcirculation in the liver (study of corrosion preparations using scanning electron microscopy)]. / Mikrocirkulace jater v norme a v experimentu. (Studium korozívních preparátu ve skanovacím elektronovém mikroskopu).

Liver vascular bed of rat is reported to be studied by scanning electron microscopy of corrosive sections. These were prepared with commercially produced methylmetacrylate monomer which was prepolymerized and injected into the prewashed and fixed vascular bed. The injections have been provided through the left cardial ventricle or directly into the liver vessels (hepatic artery, portal vein or hepatis veines). Samples from injected liver were preleved with subsequent corrosion in 40% KOH. The obtained corrosion casts were sectioned using the operational microscope and modified to be observed by scanning electron microscope Tesla BS 300. Normal liver rat pattern has been studied by scanning electron microscopy on the level of angioarchitectonics of vascular bed. The interrelation between branches of hepatic artery and portal vein was documented including their connection with evacuative venous system. In the second part of work the same method was used to study changes of configuration of liver vascular bed in rat through the early and later phases of liver regeneration after partial hepatectomy.

[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.