[Show abstract][Hide abstract] ABSTRACT: Introduction:
The risk of over-treatment in low-advanced PTC stages has prompted clinicians to search for new reliable prognostic factors. The presence of BRAF mutation, the most frequent molecular event in PTC, seems to be a good candidate. However, there is still lack of randomised trials and its significance has been proved by retrospective analyses, involving a large group of patients. The question arises whether this factor is useful in smaller populations, characterised for specialised centres. Thus, the aim of the study was to evaluate the use of BRAF mutation as a potential predictive marker in PTC patients.
233 PTC subjects treated between 2004-2006, were retrospectively analysed. Stage pT1 was diagnosed in 64.8% patients and lymph node metastases in 30.9%. Median follow-up was 7.5 years. BRAFV600E mutation was assessed postoperatively in all cases.
BRAF V600E mutation was found in 54.5%. It was more frequent in patients > 45 years (p=0.0001), and associated with larger tumour size (p=0.004). Patients with tumours <= 10 mm were over-represented among BRAF negative population (p=0.03). No association between BRAF mutation and other clinicopathological factors was observed. BRAF status was associated neither with relapse nor with disease-free survival (DFS) (p=0.76). Nodal status, extrathyroidal invasion and tumour size significantly influenced DFS.
The risk of PTC recurrence is mainly related to the presence of lymph node metastases and extrathyroidal invasion, whereas no impact of BRAF V600E mutation has been demonstrated.
PLoS ONE 07/2015; 10(7):e0132821. DOI:10.1371/journal.pone.0132821 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background:
Disturbed glucose metabolism, particularly in diabetes type 2 (DM2), may result in advanced glycation end product (AGE) formation. One of the possible targets for this reaction is lipofuscin (LF), an intracytoplasmic garbage presumed to be a marker of physiologic and preterm aging of cells. The study was performed to seek for a relationship between AGE and LF in cardiocytes of the failing hearts.
Material and methods:
Archived tissue samples from 136 hearts explanted before transplantation (in 14 pts. with ischemic cardiomyopathy (CM) and DM2; 8 pts. with dilated CM and DM2; 67 non-diabetic pts. with ischemic CM; 47 non-diabetic pts. with dilated CM), 14 autopsy cases with DM2, and 20 heart donors (control group) were involved in the study. Immunohistochemical localization of AGE was applied. The coexistence of lipofuscin and AGE was studied by LF autofluorescence in AGE-positive slides.
LF granules inside AGE deposits were present in all studied groups with varying frequencies, but the differences were non-significant. LF granules joined significantly with dispersed patterns of AGE i.e. diffuse and mixed, whereas coincidence of LF and AGE forming granular pattern was rare.
We demonstrate that LF may belong to the components of the AGE deposits. The frequency of this phenomenon is dependent on the AGE dispersion grade, but neither on diabetes nor cardiomyopathy presence.
[Show abstract][Hide abstract] ABSTRACT: Non-enzymatic coupling of protein and lipid cellular structures with glucose leading to the formation of advanced glycation end products (AGE) plays a role in aging and the development of diabetic complications, but its contribution to myocardial pathology is unclear. We aimed to assess the role of heart failure on AGE formation in patients with or without diabetes mellitus type 2 (DM2).
Heart tissue specimens from 136 patients undergoing transplantation were grouped as follows: 14 cases of ischemic cardiomyopathy (ICM) and DM2, 8 cases of dilated cardiomyopathy (DCM) and DM2, 67 cases of ICM without DM2, and 47 cases of DCM without DM2. Fourteen heart samples were from the autopsies of patients with DM2 without heart disease, and 20 heart samples were from organ donors in whom the heart was wasted. AGE deposits were localized immunohistochemically counted using a semiquantitative scale and characterized by their staining pattern.
Positive staining was present in all samples from both cardiomyopathy groups with DM2, in 71% of healthy hearts from the DM2 subjects, in 51% of ICM non-diabetic hearts, and in 38% of DCM non-diabetic hearts, and in only 15% of the organ donors. Mixed-diffuse and granular AGE patterns were characteristic for DM2, while a diffuse pattern was more frequently observed in heart failure patients without diabetes. The semiquantitative results supported increased AGE accumulation in patients with DM2 and/or cardiomyopathy.
The amount of AGE in cardiomyocytes increases significantly in both diabetes and heart failure, with a staining pattern typical for each condition.
Annals of transplantation: quarterly of the Polish Transplantation Society 06/2012; 17(2):53-61. · 1.26 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Antiapoptotic as well as replacement and proliferative mechanisms take place in the myocardium in dilated cardiomyopathy (DCM) and ischemic heart disease (IHD). We sought to estimate antiapoptotic, proliferative and replacement activities in cardiomyopathies.
The study groups included seven hearts with DCM and eight with IHD, which had been explanted at the time of transplantation. The comparator group consisted of cases of myocardial hypertrophy and the control group, donor fragments.
Antiapoptotic and proliferative responses were determined immunohistochemically as Bcl-2 and Ki67 expression by semiquantitative assessment of the intensity of staining. We also measured and statistically analyzed the integrative morphometric measurements of the fraction of fibrosis area, the nucleosarcoplasmic ratio, and cardiocyte diameter.
No Bcl-2 expression was observed in the controls. The strongest reaction was seen in the DCM group, then in the IHD, and in the comparator group of myocardial hypertrophy. Proliferative activity was seen only in endocardial and interstitial fibroblasts in DCM and IHD cases. The cardiocyte diameter showed no statistical association between myocardial hypertrophy and IHD, or IHD and DCM, whereas the nucleosarcoplasmic ratios were significantly different from control groups for all comparisons. Myocardial fibrosis showed the highest values in DCM and IHD. Discriminant analysis showed the value of interstitial fibrosis and cardiocyte diameter to categorize the analyzed groups.
Antiapoptotic Bcl-2 activity seemed to play an important role in cardiocyte preservation, while proliferative activity was resticted to interstitial connective tissue cells as a replacement process. Myocardial Bcl-2 expression, the extent of myocardial fibrosis, and cardiocyte diameter may serve as additional diagnostic tools to differentiate cardiomyopathies.
[Show abstract][Hide abstract] ABSTRACT: Morphometric studies based on the measurement of cardiocyte nuclei have focused on progressive hypertrophy rather than shape, which is a deciding factor for the diagnosis of hypertrophy in myocardial diseases. The aim of this research was to demonstrate how the digital morphology of cardiocyte nuclei change correlated with the type of myocardial pathology.
The study groups encompassed 7 hearts with dilated cardiomyopathy (DCM) and 8 hearts with ischemic heart disease (IHD) which were explanted. A comparative group consisting of myocardial hypertrophy was contrasted with a control group of donor heart fragments. Cardiocyte nuclei were evaluated morphometrically on histologic slides. We calculated the nuclear area, length, breadth, perimeter, roundness, elongation, fullness factors, and nuclear chromatin mean gray level. The results were subjected to discriminant analysis.
All karyometric measurements analyzed by backward discriminant analysis showed only 2 powerful factors: nuclear breadth and chromatin mean gray level. The Mahalanobis distance showed the proximity of control and hypertrophy groups, whereas differences between IHD and DCM were nonsignificant.
The lack of karyometric differences between IHD and DCM suggested a common morphologic response for long-lasting progressive injury. The main morphologic differences were dependent on nuclear chromatin activity/stainability and nuclear breadth, suggesting darker and thinned nuclei in normal and adaptative stages and irregular brighter nuclei in cardiomyopathies.
[Show abstract][Hide abstract] ABSTRACT: Hyperglycemia intensifies nonenzymatic glucose coupling to tissues, resulting in myocardial stiffness and formation of advanced glycation end products (AGE). The aim of this study was to assess seeking AGE in the myocardium from patients with type 2 diabetes (DM2) subjected to orthotopic heart transplantation (OHT), seeking to establish whether AGE play a role in the development of cardiomyopathies leading to OHT.
The 2 studied groups consisted of 11 hearts explanted from patients with ischemic cardiomyopathy+DM2 (ICM+DM2, 55 +/- 6.5 years) and 8 from dilated cardiomyopathy+DM2 (DCM+DM2, 49.6 +/- 4.5 years). Comparative subgroups were composed of nondiabetic explanted hearts, 41 with ICM (52.8 +/- 5.8 years) and 41 with DCM (52.7 +/- 4.2 years). All patients were males.
We examined immunohistochemical localization of AGE using a semiquantitative scale of reaction intensity in cardiomyocytes, fibroblasts, capillaries, arterioles, and arteries. Additionally, we calculated the scores for cardiocytes (AGE(Cardiocyte)) and all left ventricular components (AGE(LV)).
The cytoplasmic AGE deposits in cardiomyocytes were predominantly diffuse-granular in DM2 groups, whereas nondiabetic groups showed a lack of a reaction or a diffuse pattern. There were no differences in the reaction intensity between the 2 studied groups, or 2 comparative groups. All myocardial constituents showed higher AGE intensity in DM2 than nondiabetic groups. Only in the ICM+DM2 group did the DM2 duration correlate with AGE staining in selected myocardial layers and with AGE(Cardiocyte) and AGE(LV).
The presence of AGE in the hearts of patients requiring transplantation was related to the duration of DM2. The deposition of AGE in left ventricular myocardium was enhanced by DM2 particularly in patients with ICM.
[Show abstract][Hide abstract] ABSTRACT: Cardiocyte hypertrophy is accompanied by polyploidy, seen as a decrease in chromatin density in the enlarged nucleus. Repeated biopsies of a transplanted heart offer the possibility of a dynamic evaluation of these phenomena. The aim of this work was an evaluation of cardiocyte nuclear chromatin density in transplanted hearts during long-term follow-up.
The material encompassed myocardial biopsy specimens taken during the first week, first month, and then on an annual basis up to 10 years after surgery. Only biopsy specimens with no rejection were considered (grade "0" International Society for Heart and Lung Transplantation [ISHLT] 122 biopsy specimens). The control group consisted of 7 donor heart specimens. We evaluated the optical density-mean gray level-of cardiomyocyte nuclear chromatin. We determined correlations of this index with the nuclear area, and with left ventricle ultrasound measurements, using correlation analysis.
The chromatin mean gray level decreased with time, correlating positively with interventricular septum thickness, left ventricle posterior wall diameter, and left ventricular mass. Analysis of individual periods showed a significant positive correlation of the mean grey level with the cardiocyte nuclear surface in year 3, 4, and 9 after transplantation, thereby suggesting the occurrence of polyploidy at those times. The significant negative correlation of these values (1 week and 1 year) indicated normalization of early cardiocyte hypertrophy.
With the passage of time chromatin condenses, leading to pyknosis. The activity of cardiocyte chromatin correlated with left ventricular hypertrophy. Compensatory cardiomyocyte polyploidy is a periodical phenomenon.
[Show abstract][Hide abstract] ABSTRACT: Morphometric publications based on the measurement of cardiocyte nuclei indicated their progressive hypertrophy ignoring, however, their shape, which is a deciding factor for the microscopic-based diagnosis of hypertrophy. We sought was to demonstrate how the shapes of cardiocyte nuclei change over time and correlate them with the thickness of the interventricular septum, (IVS) the biopsy site.
We evaluated myocardial biopsies taken in the first week, first month, and then annually until posttransplant year 10. Only biopsies with no rejection were considered: grade "0" ISHLT (122 biopsies). The control group encompassed fragments from seven donor hearts.
Cardiomyocyte nuclei were evaluated morphometrically. We calculated the length, breadth, perimeter, roundness, elongation, and fullness factors for correlation with the IVS thickness, and selected indices. The relationships between karyometry and IVS thickness (measured by ultrasound) as well as time were calculated by Spearman's correlation test.
Among the examined indices, only nuclear length did not correlate significantly with follow-up time. Among the remaining indices, the strongest correlations with time were observed with regard to breadth (r = 0.214), perimeter (r = 0.150), roundness (r = -0.06) and fullness (r = 0.06), and finally elongation (r = 0.02). The decreasing thickness of the interventricular septum (r = -0.31) showed a weak correlation only with the cardiocyte nuclear length (r = -0.05).
Graft aging imitates hypertrophy inasmuch as cardiocyte nuclei become wider despite the decreased thickness of the interventricular septum. Therefore, karyometric measurements do not reflect myocardial morphology.
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to assess the effects of early thymoglobulin administration on cardiocyte nuclear status in orthotopic heart transplant (OHT) recipients.
We investigated endomyocardial biopsies (EMBs) from 31 OHT recipients and 10 control cases. OHT patients were divided into the standard group who were treated without thymoglobuline; an ATG group who received thymoglobulin electively, and a standard+ATG group who were administered thymoglobulin upon a drop in renal function. We evaluated only EMBs obtained at 1 and 4 weeks after OHT showing no significant rejection (ISHLT grades 0 to 1B). The morphometric studies were performed using a computerized, automated Quantimet image analysis system. Overall, 1750 cardiocyte nuclei were quantitated for area, length, breadth, perimeter, chromatin median grey level, and fullness factor. Statistical analysis was performed using the Mann- Whitney U test, the Wilcoxon test, and discriminant analysis.
All OHT groups showed significantly higher values (indicating nucleus enlargement) than the control group. All factors suggesting myocardial hypertrophy were significantly higher in the standard group; however, they decreased significantly with time. In contrast, the nuclear geometric parameters were significantly lower and stable throughout the study in the ATG group. The results of the standard+ATG group were intermediate, and their normalization as incomplete at the week 4 examination. Discriminant analysis revealed the closest Mahalanobis distance between control and ATG groups both at and weeks 1 and 4 after OHT.
Thymoglobulin administered early after surgery protected cardiocyte hypertrophy in heart transplant recipients, mitigating graft ischemic damage.
[Show abstract][Hide abstract] ABSTRACT: Remodeling taking place in transplanted myocardium leads to a change in the number of cardiocytes. Ultrasound measurements and biopsy evaluation should reflect their loss and compensation. We sought to evaluate the morphology of the transplanted heart upon long-term follow-up.
Myocardial biopsies were obtained in the first week, first month, and then annually for 10 years from transplantation that did not show rejection (grade "0" ISHLT, 122 biopsies). The control group encompassed 7 donor heart fragments. Proliferation in biopsies was evaluated with Ki67 (M7240, DAKO), cardiocyte hypertrophy by measuring their diameter, the surface area of the nuclei, nuclear-sarcoplasmic index, and stromal fibrosis evaluated as the surface area fraction. Ultrasound measurements included diastolic thickness of the interventricular septum, posterior wall of the left ventricle, and left ventricular mass. The correlation of measurements with time from transplantation was evaluated using Spearman's test.
A positive Ki67 reaction was observed in fibroblasts and endothelial cells. The increased cardiocyte nuclear area correlated with the time elapsed since transplantation (r = 0.2; P < .05) with a simultaneous decrease in cardiocyte thickness (r = -0.3; P < .05), without changes in the nuclear-cytoplasmic index (r = 0.02; P > .05). Stromal fibrosis also increased (r = 0.1; P < .05). Ultrasound measurements of the left ventricle showed a decreased tendency with the passage of time (r = -0.2 to -0.3; P < .05).
A transplanted heart does not undergo hypertrophy but rather fibrous atrophy with apparent compensatory hypertrophy of the cardiomyocytes.
[Show abstract][Hide abstract] ABSTRACT: Apoptotic mechanisms take place in cardiocyte death during acute heart graft rejection and remodeling by the mitochondrial pathway. This process is suppressed by Bcl-2 protein. Besides that, knowledge about cardiocyte antiapoptotic responses after heart transplantation is scanty. We sought to estimate Bcl-2 expression in the absence of rejection.
The study group included endomyocardial biopsies taken at 1 week, 1 month, 1 through to 10 years after heart transplant, which showed rejection grade "0"; the control group were donor heart fragments.
Bcl-2 expression was determined immunohistochemically by NP030 antibody (DAKO) and Envision-DAB. The intensity of staining was assessed semiquantitatively.
No Bcl-2 expression was seen in the controls; in the posttransplant groups, the significantly strongest sarcoplasmic reaction was observed at 1 week after heart transplant. Thereafter, the reaction decreased, and was weakest in the 3- and 5-year groups. From this time, Bcl-2 expression increased albeit without statistical significance. The intensity of the reaction showed no correlation with the time elapsed from heart transplant (Spearman r = 0.05; P > .05).
The expression of antiapoptotic Bcl-2 protein occurs during the entire posttransplant period, being probably a preservative and adaptative response.
[Show abstract][Hide abstract] ABSTRACT: Distribution and staining of the nuclear chromatin are sensitive indicators of changes in physiology and pathology of cells. However, their use in heart transplant recipients is rare. The aim of this study was to compare cardiomyocyte status in heart transplant recipients suffering from moderate acute cellular rejection and subjects without signs of active cellular rejection.
One hundred twenty-nine endomyocardial biopsy samples from 43 heart transplant recipients (no later than 6 months after surgery) were evaluated. Overall, 3235 cardiomyocytic nuclei were analyzed using the Quantimet image analysis system to assess the mean gray level: 1584 nuclei were found in biopsies without signs of rejection or with nonsignificant rejection (ISHLT grades 0, 1A, and 1B), whereas the remaining 1651 nuclei were measured from biopsies showing ISHLT grade 3A (significant rejection). Additionally, the chromatin distribution was assessed in all eligible nuclei.
The mean gray level was markedly increased in nuclei from biopsy samples with significant rejection (182.6 vs 112.5, P < .001, Mann-Whitney U test). Moreover, the analysis of chromatin distribution revealed significantly more frequent chromatin marginalization and irregular distribution in rejecting subjects (P < .001, chi-square test).
Inflammatory stimulation of cardiomyocytes during acute cellular rejection of the transplanted heart influences the chromatin distribution in the nuclei, which may have an additional value, when assessing the severity of rejection.
[Show abstract][Hide abstract] ABSTRACT: Histology of the soft palate and uvula in snorers and patients with OSA syndrome has been a subject of investigation of many authors. In majority of specimens the hypertrophy of the salivary glands as well as congestion and dilation of the thin-walled vessels were observed. Some of the samples presented atrophy of the muscle bundles. Also, the inflammatory changes as lymphocytic infiltrations were proven. In each case adults served as a control group. In this study the histologic analysis of the soft palate and uvula samples from patients suffering from snoring and OSA syndrome was performed. The uvula samples from the newborn who died on the first day of their lives were chosen as a control group. The choice of such a group excluded the influence of vibration force on the soft palate structure. Comparisons among the groups were made by Fisher exact two-tailed test. The muscular atrophy was observed only in patients with airway disturbances. No case of neonatal tissue sample with this pathology was found. Less dilation and congestion of the blood vessels were observed in the newborn group. Significantly more frequent superficial salivary glands localized between the muscle bundles and epithelium were found in the OSA and snoring patients. Our results showed distinct differences between the tissues of the patients with airway disturbances and the control group. These differences may be caused by the influence of the vibration on the soft palate and uvula, but on the other hand they may be the reason for excessive flaccidity of these structures and disturbances occurring during sleep.
Otolaryngologia polska. The Polish otolaryngology 02/2005; 59(1):13-9.
[Show abstract][Hide abstract] ABSTRACT: The purpose of this study was to characterize normal epithelium, precancerous lesions and laryngeal squamous cell carcinoma (SCC) with the use of nuclear morphometry. The relationship between the morphometric parameters of the cell nuclei and the pathological lesions was analyzed. In addition, a morphometric comparison was performed among normal epithelium, precancerous lesions, and cancer of the larynx.
Our research involved 12 patients after total laryngectomy. All the postoperative histological specimens consisted of laryngeal SCC, precancerous lesions (dysplasia), keratosis with mild dysplasia, and normal epithelium. The cell nuclei of the SCC, dysplasia, keratosis, and the basal and superficial layers of the normal epithelium were measured. The nuclear parameters included area, equivalent diameter, length, breadth, perimeter, mean and integrated grey levels, convex area and convex perimeter, roundness, aspect ratio and fullness ratio. For each case, 500 nuclei were studied using a Polyvar Reicher microscope connected to a Quantimet 500 computer system.
The relationship between karyometrical parameters and pathological lesions was found to be statistically significant. We observed the largest nuclei in keratosis lesions and the smallest in basal layers of normal epithelium. In some cases there were no significant differences between dysplasia nuclei and the superficial layers of normal epithelium.
Karyometrical parameters correlate with morphometrical lesions in the larynx and may be useful in the prediction of laryngeal neoplasm.
Medical science monitor: international medical journal of experimental and clinical research 07/2004; 10(6):CR241-5. · 1.43 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The authors assessed morphometrically the nuclei shape of the normal mucosa, precancerous lesions (dysplasia, keratosis) and laryngeal cancer in the postoperative samples of 12 patients treated in the Laryngology Department Silesian Medical Academy in Zabrze. Five slides from the same patient, each 4 um thick were assessed. The histologic samples were measured by system Quantimet 500 + Color Option with magnification 2100x. Roundness, aspect ratio and fullness ratio were evaluated. The relationship among the morphologic lesions using shape descriptors was assessed. In some cases there were no significant differences between the nuclei of mild dysplasia cells and the cells of the normal epithelium. The results indicate significant karyometric shape differences of the normal mucosa, precancerous lesions and cancer of the larynx.
Otolaryngologia polska. The Polish otolaryngology 02/2003; 57(6):855-9.
[Show abstract][Hide abstract] ABSTRACT: The aim of the study was the characterization of calcification in the leaflets of a cryopreserved and alive heart valve depending on the diagnosed pathologic process. Sheep antibiotic sterilised and cryopreserved biological valves were implanted in tricuspid position in young sheeps for one year period. After this time the valves removed and studied morphologically. The control group consisted of 7 intact valves, the comparative group, so called group of valves after the processing antibiotic sterilization and cryopreservation consisted of 7 valves after mentioned procedures. Histological investigations were based on paraffin sections, calcium deposits were stained von Kossa technique. The measured values included integrative parameters as: 1. area fraction, 2. number of calcifications per area, 3. anisotropy. Conclusions: 1. A process of initial processing, sterilization and cryopreservation of biological valve increases a number of microcalcifications. 2. Cryopreserved biological valves explanted after one-year implantation into an animal in a tricuspid position possess fine calcifications and calcification foci. A number and size of fine calcifications decreases together with an intensification of degeneration and regressive processes of the connective tissue, especially in hyalinization. Hyalinization of the biological valve tissue seems to be favorable for a valve durability and as a pathological process decreasing calcification. 3. Mathematic analysis of morphometric features defining density and structure of calcifications indicate similarities among cryopreservation and initial processing groups, hyalinization, inflammation, whereas in a group of calcification foci, the similarity can be noticed between inflammation and hyalinization group.
Annals of transplantation: quarterly of the Polish Transplantation Society 02/2003; 8(1):45-54. · 1.26 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The aim of the study was an evaluation of viability and damage of biological cells sterilized with antibiotics and cryopreserved a year after the implantation. Sheep antibiotic sterilised and cryopreserved biological valves were implanted in tricuspid position in young sheeps for one-year period. After this time the valves removed and studied morphologically. The control group consisted of 7 intact valves, the comparative group, so called group of valves after the processing antibiotic sterilization and cryopreservation consisted of 7 valves after mentioned procedures. Histologic investigations were based on paraffin sections of formalin-fixed valve cusps, stained with H&E and Masson trichrome. Valve viability was assessed using intravital staining with fluoresceine diacetate, whereas damaged cells were visualized by intravital staining with neutral red. Additionally, the ultrastructural studies were performed. The viability and ultrastructural results were compared with the pathologic process in the valve. Conclusions: 1. preliminary preparation with antibiotic sterilisation and cryopresrevation induces valve leaflet oedema and degenerative ultrastructural processes, colliquative cell necrosis and apoptotic morphology of a part of valve cells; 2. cryopreserved and antibiotic sterilised pulmonary valve after one-year implantation behaves lining cells, but ultrastructural changes indicates many degenerative phenomena in smaller degree than after preparation, sterilization and cryopreservation. Histopathologically the degenerative changes were prevalent.
Annals of transplantation: quarterly of the Polish Transplantation Society 02/2003; 8(1):70-8. · 1.26 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The aim of the study was morphometric and mathematic analysis of calcification profiles present in the leaflets of a cryopreserved and alive heart valve depending on the diagnosed pathologic process. Sheep antibiotic sterilised and cryopreserved biological valves were implanted in tricuspid position in young sheeps for one-year period. After this time the valves removed and studied morphologically. The control group consisted of 7 intact valves, the comparative group, so called group of valves after the processing antibiotic sterilization and cryopreservation consisted of 7 valves after mentioned procedures. Histologic investigations were based on paraffin sections of formalin-fixed valve cusps, stained with H&E and Masson trichrome, calcium deposits were stained von Kossa technique. The measured values included: 1. area and equivalent diameter, 2. length, 3. breadth, 4. perimeter, 5. elongation, 6. roundness, 7. fullness coefficient. Conclusions: 1. A process of initial processing and cryopreservation of biological valve increases a dimension and disturbs a shape of microcalcifications. 2. Cryopreserved biological valves explanted after one-year implantation into an animal in a tricuspid position possess microcalcifications and calcification foci. The size of microcalcifications decreases together with an intensification of degenerative processes of the connective tissue, especially in hyalinization. Hyalinization of the biological valve tissue seems to be favorable for a valve durability and as a pathological process decreasing calcification. 3. Mathematic analysis of morphometric features defining differences in size and shape of each calcification indicate morphologic and morphometric autonomy of calcifications, characteristic for the analyzed group of valve pathologic changes.
Annals of transplantation: quarterly of the Polish Transplantation Society 02/2003; 8(1):55-69. · 1.26 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The assessing of heart valve viability is critical for the preparing of viable and implantable biological value. Non-viable valves should be cross-linked with various agents whereas viable tissues are suitable for cryoprotection. Cell viability assessment should be rapid, simple, easy and low cost.
A simple method, simultaneous fluorescent staining of both viable and damaged valve cells was described. The material was consisted of 2 groups: a) with ischemic time below 3 hours (5 swine valves), and b) with ischemic time over 3 hours (5 swine valves). The supravital staining of leaflet fragments was conducted with single solution of fluorescein diacetate (living cells staining) and propodium iodide (damaged cells staining).
The simultaneous staining predominantly showed green fluorescence typical for living cells and few red damaged cells in the short ischemic time group, whereas the second group showed the predominance of the red stained damaged cells.
The presented one-step supravital staining method is rapid and simple it allows differentiation of the viability of the valve cells, so it may be of value for the routine evaluation of cadaveric valve homografts.
Medical science monitor: international medical journal of experimental and clinical research 01/2001; 7(3):461-3. · 1.43 Impact Factor