A comprehensive morphometric analysis of the internal thoracic artery with emphasis on age, gender and left-to-right specific differences.

AIMS OF THE STUDY: The aim of this analysis was the morphometric description of the internal thoracic artery (ITA) with an emphasis on age, gender and left-to-right specific differences, as well as on age and atherosclerosis related changes of the elastic skeleton. METHODS: Forty eight arteries were obtained during forensic autopsies from 32 persons who had died of non-vascular causes. The following morphometric parameters were analyzed: thickness of the intima, the medial layer and the wall, the intima-to media-ratio and the elastic skeleton parameters. RESULTS: The intima thickness increases significantly with aging (ANOVA F=34.061, p⟨0.001), as does the intima-to-media ratio (ANOVA F=10.831, p⟨0.001). With aging, there is a significant increase in the thickness of the media (F=56.519; p⟨0.001) and of the wall (F=34.094; p⟨0,001). There is a significant increase in the media thickness during the development of atherosclerosis in the ITA (ANOVA F=11.848, p⟨0.001). No significant difference was found when these data were analyzed based on the left-to-right principle or depending on gender of the patients. However, the analysis of the elastic skeleton parameters indicated that the combined effects of aging, atherosclerosis and male gender lead to the degeneration of the elastic skeleton of the ITA. CONCLUSION: The grade of atherosclerosis gradually increases with aging as shown by morphometric analysis. The increase in the medial layer thickness suggests the potential for positive remodeling of the ITA during aging and atherosclerosis. The left/right position has no influence on morphometric parameters of the ITA, while male gender affects parameters of the elastic skeleton.

Skin and sural nerve biopsies: ultrastructural findings in the first genetically confirmed cases of CADASIL in Serbia.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited vascular disorder caused by Notch3 gene mutations. The main histopathological hallmark is granular osmiophilic material (GOM) deposited in the close vicinity of vascular smooth muscle cells (VSMCs). The authors report the first 7 ultrastructurally and genetically confirmed cases of CADASIL in Serbia. Samples of skin and sural nerve were investigated by transmission electron microscopy. GOM deposits were observed around degenerated VSMCs in all the skin biopsies examined. Sural nerve biopsies revealed severe alterations of nerve fibers, endoneurial blood vessels with GOM deposits, endoneurial fibroblasts, and perineurial myofibroblasts. Total genomic DNA was extracted from peripheral blood leukocytes, and exons 2-6 of the Notch3 gene were amplified by PCR and subsequently sequenced. Four different mutations in exons 2 (Cys65Tyr), 3 (Gly89Cys and Arg90Cys), and 6 (Ala319Cys), which determine the CADASIL disease, were detected among all described patients. A novel missense mutation Gly89Cys involving exon 3 was detected. Due to the difficulties in the determination of the Notch3 mutations, these data suggest that electron microscopic analysis for GOMs in dermal vessel wall provides a rapid and reliable screening method for this disease.

[Mitochondrial myopathy, encephalopathy, lactate acidosis with stroke-like episodes syndrome (MELAS): a case report].

INTRODUCTION: Mitochondrial encephalopathy, lactacidosis and stroke-like episodes (MELAS) represent a multisystemic dysfunction due to various mutations in mitochondrial DNA. Here we report a patient with genetically confirmed MELAS. CASE OUTLINE: A patient is presented whose clinical features involved short stature, easy tendency to fatigue, recurrent seizures, progressive cognitive decline, myopathy, sensorineural deafness, diabetes mellitus as well as stroke-like episodes. The major clinical feature of migraine type headache was not present. Neuroimaging studies revealed signs of ischemic infarctions localized in the posterior regions of the brain cortex. Electron microscopy of the skeletal muscle biopsy showed subsarcolemmal accumulation of a large number of mitochondria with paracristal inclusions in the skeletal muscle cells. The diagnosis of MELAS was definitively confirmed by the detection of a specific point mutation A to G at nucleotide position 3243 of mitochondrial DNA. CONCLUSION: When a relatively young patient without common risk factors for ischemic stroke presents with signs of occipitally localized brain infarctions accompanied with multisystemic dysfunction, MELAS syndrome, it is necessary to conduct investigations in order to diagnose the disease.

Micro- and nanostructures of iridescent wing scales in purple emperor butterflies (Lepidoptera: Apatura ilia and A. iris).

Apatura ilia (Denis and Schiffermüller, 1775) and A. iris (Linnaeus, 1758) are fascinating butterflies found in the Palaearctic ecozone (excepting the north of Africa). The wings of these insects are covered with a great number of two types of scales positioned like roof tiles. Type I scales are on the surface, while type II scales are situated below them. The structural color of the type I scales is recognized only on the dorsal side of both the fore and hind wings of the males of the aforementioned species. Both types of scales are responsible for pigment color of the wings, but iridescence is observed only in the type I scales. The brilliant structural color is due to a multilayer structure. The features of the scales, their dimensions and fine structure were obtained using scanning electron microscopy. Cross sections of the scales were then analyzed by transmission electron microscopy. The scales of the "normal" and clytie forms of A. ilia have a different nanostructure, but are of the same type. A similar type of structure, but with a different morphology, was also noticed in A. iris. The scales of the analyzed species resemble the scales of tropical Morpho butterflies.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy.

INTRODUCTION: Fast and precise diagnostics of the disease from the large group of adult leukoencephalopathy is difficult but responsible job, because the outcome of the disease is very often determined by its name. Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is caused by the mutation of Notch 3 gene on chromosome locus 19p13. Beside the brain arterioles being the main disease targets, extracerebral small blood vessels are affected by the pathological process. Clinically present signs are recurrent ischemic strokes and vascular dementia. CADASIL in its progressive form shows a distinctive pattern of pathological changes on MRI of endocranium. The diagnosis is confirmed by the presence of granular osmiophilic material (GOM) in histopathological skin biopsies. CASE REPORTS: Two young adult patients manifested ischemic strokes of unknown etiology, cognitive deterioration, migraine and psychopathological phenomenology. MRI of endocranium pointed on CADASIL. Ultrastructural examination of skin biopsy proved the presence of GOM in the basal lamina and near smooth muscle cells of arteriole dermis leading to CADASIL diagnosis. The presence of GOM in histopathological preparation is 100% specific for CADASIL. The patients were not searched for mutation in Notch 3 gene on chromosome 19, because some other leukoencephalopathy was disregarded. CONCLUSION: Suggestive clinical picture, distinctive finding of endocranium MRI, the presence of GOM by ultrastructural examination of histopathological skin biopsies are sufficient to confirm CADASIL diagnosis.

The internal thoracic artery as a transitional type of artery: a morphological and morphometric study.

Coronary artery by-pass grafting (CABG) with arterial grafts is widely accepted as the procedure of choice in the treatment of coronary ischemic disease. It brings back focus on morphological studies of arteries used as conduits in this procedure. One of the most frequently used CABG grafts is the internal thoracic artery with an excellent graft prognosis and patency rate. The aim of the study was a detailed morphological and morphometric description of the internal thoracic artery with an emphasis on its basic histological structure and its changes in aging and atherosclerosis. Therefore, 42 full-length arteries were obtained during forensic autopsies from 27 persons, aged between 20 and 81 years, who had died from non-vascular causes. The arteries were classified into three different age groups. Analysis of the serial arterial segments has shown that the internal thoracic artery is an artery of the transitional type whose media is organized into two layers: the internal, muscular layer and the external layer with spirally oriented elastic lamellae and smooth muscle cells in between. The number of elastic lamellae progressively decreases throughout the length of the examined arteries. As opposed to previous assumptions, we have proven that the grade of atherosclerosis is independent of the number of elastic lamellae in the external media. Perfectly formed elastic lamellae are not a persistent feature of the internal thoracic artery, as previously claimed. We have confirmed that the thickness of elastic lamellae decreases, while the number and the size of their fenestrations steadily increase with aging.

Endothelial keratoplasty without descemet's membrane stripping: histologic and ultrastructural findings.

AIM: To report histologic and ultrastructural findings of endothelial keratoplasty (EK) performed without Descemet's membrane stripping. METHODS: Clinical techniques, histology, and transmission electron microscopy. RESULTS: A 55-year-old woman was referred to us, after 2 unsuccessful penetrating keratoplasties (PKs), for pseudophakic bullous keratopathy. An 8.0-mm EK without Descemet's membrane stripping was performed, and clarity was restored to the failed penetrating regraft. A year later, the lamellar graft failed, and a third PK was performed for intractable corneal edema. Light microscopy of a semi-thin section of the trephined corneal button showed both the recipient's and the donor's Descemet's membrane, the well-preserved structure of the full-thickness graft, and marked edema of the adherent stromal carrier of the endothelial transplant. The host endothelium was absent at the interface, and the donor endothelium was atrophic. Electron microscopy revealed regularity and even spacing of collagen fibrils as well as quiet keratocytes on both sides of Descemet's membrane at PK-EK interface. CONCLUSION: These findings suggest a lack of proliferation and hypercellular scarring, and offer further support to the already proven merits of EK.

Use of recombinant factor VIIa in the treatment of massive retroperitoneal bleeding due to severe necrotizing pancreatitis.

BACKGROUND: Recently, a growing number of case reports and case series have suggested that the use of recombinant activated factor VII (rFVIIa) may be effective in treatment of patients with non-hemophilic acquired coagulopathy not responding to conventional treatment such as major surgery, major trauma, sepsis, necrotizing pancreatitis and bleeding due to cerebral arteriovenous malformations. CASE REPORT: We presented a septic patient with massive, life-threatening bleeding caused by retroperitoneal necrosis, due to severe acute necrotizing pancreatitis. As conservative treatment (blood, plasma, cryoprecipitates and platelet transfusions) failed to induce cessation of bleeding, the patient was urgently operated on. In spite of usual procedures of surgical hemostasis (ligation, suture, thermocauterisation, fibrin glue, temporary tamponade), hemorrhage could not be stopped. The patient manifested the signs of hypothermia and metabolic acidosis and, therefore, the decision was made to use recombinant activated factor VII (Novo Seven). The application of rFVIIa resulted in significant discontinuation of hemorrhage, restoration to normal blood count as well as other relevant coagulation parameters. CONCLUSION: Although application of rFVIIa is still in the initial clinical phase, and the experience is based mainly on uncontrolled series as well as on individual observations, it seems that this drug can be promising, potent and attractive adjunctive prohemostatic agent. This drug may play a beneficial role in the treatment of serious and unresponsive, "nonsurgical", life-threatening bleeding due to severe acute necrotizing pancreatitis.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy with severe factor XII deficiency.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited adult-onset microangiopathy caused by missense mutations in the Notch3gene on chromosome 19. However, common vascular risk factors may additionally modify clinical expression and progression of the disease. The role of various prothrombotic factors has also been implied. We report a case of a middle-aged man with typical clinical, neuroimaging and histological features of CADASIL, but with notably prolonged activated partial thromboplastin time. Hematological investigations revealed severe clotting Factor XII deficiency. This case illustrates that the occurrence of vascular risk factors should not be overlooked in patients with CADASIL.

[Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL)--three case reports from Serbia].

INTRODUCTION: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a hereditary microangiopathy leading to recurrent strokes and vascular dementia in young and middle-aged patients. The diagnosis of CADASIL is based on typical clinical presentation and characteristic magnetic resonance imaging (MRI) changes, and has to be confirmed by biopsy of the sural nerve, muscle and skin, as well as by genetic analysis. Mutations within the Notch3 gene were identified as the underlying genetic defect in CADASIL. CASE OUTLINE: The clinical manifestations of the first presented patient with migraine from the age of thirteen, stroke without vascular risk factors and stepwise progression of vascular dementia comprising the typical clinical picture of CADASIL, were confirmed after seven years with pathological verification. The second presented case did not satisfy the clinical criteria for CADASIL. His stroke was considered to be related with vascular risk factors--diabetes mellitus and hypertension. The aetiological diagnosis was established only when his brother without vascular risk factors presented with similar clinical manifestations. CONCLUSION: Until the development of the new neuroimaging techniques like MRI, pathologic and genetic analysis, CADASIL was considered as a rare disorder. However, the increasing number of CADASIL families has been identified throughout the world showing that this entity is usually underdiagnosed. This article presents three patients from two Serbian families with clinical suspicion of CADASIL verified by pathologic examination.

Invasive mole--case report of massive uterine destruction.

Patient with malignant Gestational Trophoblastic Neoplasm (GTN) was treated by mean of MTX-FA, MAC, EMA-CO and EMA-EP. Changes in serum human chorionic gonadotropine (beta hCG) levels and changes in ultrasonographic findings were checked weekly. Finally transabdominal hysterectomy with ovaries conservation was done and polychemotherapy administrated after the operation until three consecutive serum chorionic gonadotropine values were negative. This is a case report of Invasive mole in 32 years old patient without possibility to preserve reproductive health. GTN developed two months after spontaneous abortion in 13th week gestation. No changes in uterine structure were found during the first ultrasonographic examination. Three months after abortion and one month after GTN confirmed, massive destruction of lateral uterine wall was detected during transvaginal Doppler ultrasound examination. Resistance index of 0,366 was significantly lower than normal, with hypervascularisation in affected tissue. Serum beta hCG confirmed poor effect of polychemotherapy treatment and decision for operative treatment was made. Hystological findings after the operation confirmed malignant GTN- invasive mole. Specific changes in ultrasonographic picture could have an impact in therapy making decision and could not be refereed without the most relevant parameter such is serum human chorionic gonadotropine.

[Cardiac myxoma -- the influence of preoperative clinical presentation and surgical technique on late outcome].

INTRODUCTION: Cardiac myxomas are the most frequent primary tumours of the heart in adults, and they can be found in each of four cardiac chambers. Although biologically benign, due to their unfavourable localization, myxomas are considered "functionally malignant" tumours. Diagnosis of cardiac myxoma necessitates surgical treatment. OBJECTIVE: To analyse: 1) the influence of localization, size and consistency of cardiac myxomas on preoperative symptomatology; 2) the influence of different surgical techniques (left, right, biatrial approach, tumour basis solving) on early, and late outcomes. METHOD: From 1982 to 2000, at the Institute for Cardiovascular Diseases, Clinical Centre of Serbia, there were 46 patients with cardiac myxomas operated on, 67.4% of them women, mean age 47.1 +/- 16.3 years. The diagnosis was made according to clinical presentation, electrocardiographic and echocardiographic examinations and cardiac catheterization. Follow-up period was 4-18 (mean 7.8) years. RESULTS: In 41 (89.1%) patients, myxoma was localized in the left, while in 5 (10.9%), it was found in the right atrium. Average size was 5.8 x 3.8 cm (range: 1 x l cm to 9 x 8 cm) and 6 x 4 cm (range: 3 x 2 cm to 9 x 5 cm) for the left and right atrial myxomas, respectively. A racemous form predominated in the left (82.6%) and globous in the right (80%) atrium. Fatigue was the most common general (84.8%) and dyspnoea the most common cardiologic symptom (73.9%). Preoperative embolic events were present in 8 patients (4 pulmonary, 4 systemic). In our series: 1) different localization, size and consistency had no influence on the preoperative symptomatology; 2) surgical treatment applied, regardless of different approaches and basis solving, resulted in excellent functional improvements (63.1% patients in NYHA III and IV class preoperatively vs. 6.7% patients postoperatively) and had no influence on new postoperative rhythm disturbances (8.7% patients preoperatively vs. 24.4% patients postoperatively); 3) early (97.8%), and late survival rates (91.3%) were excellent; 4) there were no relapses during the follow-up period. CONCLUSION: Localization, size and consistency had no influence on the preoperative symptomatology. Excellent survival rate with significant functional improvement, rare postoperative complications and no recurrences, justify the applied strategies of surgical approach and tumour basis solving in our series.

The helical ventricular myocardial band of Torrent-Guasp.

We live in an era of substantial progress in understanding myocardial structure and function at genetic, molecular, and microscopic levels. Yet, ventricular myocardium has proven remarkably resistant to macroscopic analyses of functional anatomy. Pronounced and practically indefinite global and local structural anisotropy of its fibers and other ventricular wall constituents produces electrical and mechanical properties that are nonlinear, anisotropic, time varying, and spatially inhomogeneous. The helical ventricular myocardial band of Torrent-Guasp is a revolutionary new concept in understanding global, 3-dimensional, functional architecture of the ventricular myocardium. This concept defines the principal, cumulative vectors, integrating the tissue architecture (ie, form) and net forces developed (ie, function) within the ventricular mass. The primary purpose of this review is to emphasize the importance of this concept, in the light of collaborative efforts to establish an integrative approach, defining ventricular form and function by linking across multiple scales of biological organization, as explained in the ongoing Physiome project. Because one of the most important scientific missions in this century is integration of basic research with clinical medicine, we believe that this knowledge is not of merely academic importance, but is also the essential prerequisite in clinical evaluation and treatment of different heart diseases.

[Case report of ultrasonographic findings in developing choriocarcinoma]. / Prikaz slucaja razvoja horiokarcinoma dijagnosticiranog metodom ultrasonografije.

INTRODUCTION: Gestational Trophoblastic Neoplasm (GTN) develops from fetal tissue with risk of malignancy after complete hydatidiform mole of 20%. MATERIAL AND METHODS: This is a case report of progressive destruction of uterine tissue in 43 years old patient caused with developing Choriocarcinoma three months after legal abortion. Severe changes in uterine structure as multiple anechogenic foci of destruction were found during transvaginal Doppler ultrasound examination. Hypervacularisation in malignant trophoblastic tissue with low Resistance index values (0,397-0,471) were also detected during transvaginal Doppler ultrasonography. Ultrasonographic picture significantly changed in two months followed with serum human chorionic gonadotropine increasing rate. Operative treatment performed and polychemotherapy administrated. Postoperative hystological examination confirmed Choriocarcinoma. DISCUSSION: The most relevant parameter in making GTN diagnose is serum human chorionic gonadotropine level. This case of choriocarcinoma showes that ultrasonographic changes can sometimes bee more progressive than expected value of serum hCG according to those findings. Transvaginal Doppler ultrasonographic findings must be compared with changes in serum human chorionic gonadotropine values and hystological findings. CONCLUSION: Choriocarcinoma is not easy to detect ultrasonographicaly but in some cases ultrasonographic picture can be specific for the case as one of numerous ultrasonographic models choriocarcinoma can appear in.

[Cytohistological and immunohistochemical characteristics of vascular remodelling in diseases of the blood vessels].

INTRODUCTION: Vascular remodelling is an adaptive process involving the adjustment of the structure and function of blood vessels to long-term changes in haemodynamic conditions. This process leads to structural alterations within vessel walls in different cardiovascular diseases, such as hypertension, atherosclerosis, and coarctation of the aorta. OBJECTIVE: We investigated the histochemical and immunocytochemical characteristics of morphological lesions in coronary atherosclerosis and coarctation of the aorta. METHOD: Twenty-one samples of atherosclerotically modified right coronary arteries, divided into 6 segments, were analysed. We also examined 10 samples of coarctation segments, excised during surgery. The segments were stained histochemically (using orcein and alcian blue-PAS), immunocytochemically (using alpha-smooth muscle actin-alpha-SMA, vimentin, desmin, myosin heavy chains-MHC, CD3, CD45, S-100, and Proliferating Cell Nuclear Antigen-PCNA), and for electron microscopy. RESULTS: The results of our study of morphological lesions in coronary atherosclerosis demonstrated initial functional and then, in the later stages of atherosclerosis, morphological, damage to the endothelium. The preatheroma stage revealed the presence of intimal dedifferentiation of smooth muscle cells, with the expression of vimentin and alpha-SMA, and the lack of expression of desmin. Along with these changes, a huge number of foam cells of variant origin were noticed. Some of them were CD68-immunoreactive while others were both vimentin- and S-100-immunoreactive. All examined samples of the coarctation of the aorta demonstrated the presence of dedifferentiated smooth muscle cells as well as a diminution in cell numbers, followed by apoptotic smooth muscle cells, and the absence of inflammatory cells. CONCLUSION: Some foam cells develop from monocyte-macrophage lineage (CD68-immunoreactive), while others originate from smooth muscle cells (vimentin and S-100-immunoreactive). Coarctation of the aorta is characterised by a diminution in cell numbers (apoptosis) as well as their dedifferentiation from contractile to synthetic phenotype.

The helical ventricular myocardial band: global, three-dimensional, functional architecture of the ventricular myocardium.

We are currently witnessing the advent of new diagnostic tools and therapies for heart diseases, but, without serious scientific consensus on fundamental questions about normal and diseased heart structure and function. During the last decade, three successive, international, multidisciplinary symposia were organized in order to setup fundamental research principles, which would allow us to make a significant step forward in understanding heart structure and function. Helical ventricular myocardial band of Torrent-Guasp is the revolutionary new concept in understanding global, three-dimensional, functional architecture of the ventricular myocardium. This concept defines the principal, cumulative vectors, integrating the tissue architecture (i.e. form) and net forces developed (i.e. function) within the ventricular mass. Here we expose the compendium of Torrent-Guasp's half-century long functional anatomical investigations in the light of ongoing efforts to define the integrative approach, which would lead to new understanding of the ventricular form and function by linking across multiple scales of biological organization, as defined in ongoing Physiome project. Helical ventricular myocardial band of Torrent-Guasp may also, hopefully, allow overcoming some difficulties encountered in contemporary efforts to create a comprehensive mathematical model of the heart.

The origin and differentiation potential of smooth muscle cells in coronary atherosclerosis.

OBJECTIVES: To determine the phenotypical state of smooth muscle cells during the pathogenesis of an atherosclerotic lesion, and to determine the morphological state of the endothelium and the origin of foam cells. METHODS: Twenty-one samples of atherosclerotically changed right coronary arteries, which were divided into six subgroups based on the stage of atherosclerosis, were analyzed. The tissues were fixed in formalin and embedded in paraffin. Sections of 5 mum thickness were stained immunocytochemically using a labelled streptavidin-biotin/horse radish peroxidase kit (Dako, Denmark) for the identification of vimentin, alpha-smooth muscle actin, myosin heavy chains, desmin, S-100 protein, CD3, CD31, CD34, CD45, CD68 and proliferating cell nuclear antigen protein. RESULTS: The present study showed that there is first functional and then morphological damage of the endothelium in the late stages of atherosclerosis. The preatheroma stage revealed the presence of intimal changes of smooth muscle cells, with expression of vimentin and alpha-smooth muscle actin and a lack of expression of desmin, which led to a switch to a synthetic phenotype. The described changes progressed into the later stages of atherosclerosis. Along with these changes, a large number of foam cells of variant origin were observed; some of the foam cells developed from monocyte-macrophage lineage (CD68-immunoreactive) and others originated from smooth muscle cells (vimentin- and S-100-immunoreactive). The late stages of atherosclerosis development, such as the atheroma stage, include intimal changes with the formation of a lipid core (S-100-immunoreactive cells and cell necrosis), while fibrosis in the lipid core and the accumulation of collagen fibres with extreme hypocellularity are characteristics of the fibroatheroma stage.