Immunohistochemical analysis of MMP-9 and COX-2 expression in carotid atherosclerotic plaques among patients undergoing carotid endarterectomy: A prospective study.

BACKGROUND: Matrix metalloproteinase-9 protein (MMP-9) and cyclooxygenase-2 (COX-2) proteins may have a role in remodelling of atherosclerotic plaques. We analysed and compared the radiological, histological and immunohistochemical characteristics of carotid atherosclerotic plaques between symptomatic and asymptomatic patients who underwent carotid endarterectomy (CEA). METHODS: This prospective single-blinded study included 31 patients (70 [64-75] years, 58% males, 42% symptomatic) who underwent CEA and a total of 155 carotid plaque sections that were analysed. Preoperative assessment and multimodality diagnostic imaging with magnetic resonance imaging (MRI) or computed tomography angiography (CTA), histological and immunohistochemical analyses of carotid plaques including the expression of MMP-9 and COX-2 proteins were performed. RESULTS: Symptomatic and asymptomatic patients did not significantly differ in respect to preoperative characteristics. Unstable plaques were detected in 12/13 (92.3%, p = 0.020) symptomatic patients using MRI or CTA. There was no perioperative mortality and perioperative outcomes were comparable in both groups. A significantly higher expression of MMP-9 in macrophages was observed among symptomatic patients (p = 0.020). ROC curve analysis showed statistically significant associations of both the higher intensity of COX-2 staining in CD68 PG-M1 positive macrophages (area under the curve [AUC]=0.701, p = 0.014) and higher MVD (AUC=0.821, p < 0.001) within the plaque with cerebrovascular symptoms. The expression of COX-2 and the intensity of COX-2 staining in macrophages within the unstable carotid plaques detected by preoperative MRI or CTA were significantly higher (76.1% vs. 40.0%, p = 0.038; 76.2% vs. 30.0%, p = 0.01, respectively). CONCLUSIONS: Advanced non-invasive multimodality diagnostic imaging including MRI or CTA is reliable in differentiating unstable from stable carotid plaques. High expression of MMP-9 and COX-2 in macrophages within the symptomatic plaque is associated with increased risk of cerebrovascular complications. TRIAL REGISTRATION: This study has been registered at the ISRCTN registry (ID ISRCTN46536832), isrctn.org Identifier: https://www.isrctn.com/ISRCTN46536832.

The role of magnetic resonance imaging and the expression of MMP-9 protein in the analysis of carotid atherosclerotic plaques in patients undergoing carotid endarterectomy: a prospective pilot study.

Components of carotid atherosclerotic plaque can be analysed preoperatively by non-invasive advanced imaging modalities such as magnetic resonance imaging (MRI). The expression of matrix metalloproteinase-9 protein (MMP-9), which has a potential role in remodelling of atherosclerotic plaques, can be analysed immunohistochemically. The aim of the present prospective pilot study is to analyse histological characteristics and expression of MMP-9 in carotid plaques of patients undergoing carotid endarterectomy (CEA) and to investigate the correlation with preoperative clinical symptoms and MRI features. Preoperative clinical assessment, MRI imaging, postoperative histological and immunohistochemical analyses were performed. Fifteen patients with symptomatic (7/15; 47%) and asymptomatic carotid artery stenosis undergoing CEA were included. Among symptomatic patients, 5 (71%) had recent stroke and 2 (29%) had recent transient ischaemic attack with a median timing of 6 weeks (IQR: 1, 18) before the surgery. Both groups did not significantly differ in respect to preoperative characteristics. Prevalence of unstable plaque was higher in symptomatic than asymptomatic patients, although it was not significant (63% vs. 37%, p = 0.077). The expression of MMP-9 in CD68 cells within the plaque by semiquantitative analysis was found to be significantly higher in symptomatic as compared to asymptomatic patients (86% vs. 25% with the highest expression, p = 0.014). The average microvascular density was found to be higher and lipid core area larger among both symptomatic patients and unstable carotid plaque specimens, although this did not reach statistical significance (p = 0.064 and p = 0.132, p = 0.360 and p = 0.569, respectively). Our results demonstrate that MRI is reliable in classifying carotid lesions and differentiating unstable from stable plaques. We have also shown that the expression of MMP-9 is significantly higher among symptomatic patients undergoing CEA.

Endovascular Management of Superior Mesenteric Artery (SMA) Aneurysm - Adequate Access is Essential for Success - Case Report.

BACKGROUND: An aneurysm of the superior mesenteric artery (SMA) with a diameter of 2.2 cm was found incidentally on an ultrasound (US) examination in a 26-year-old woman. The only known risk factor was an intracranial aneurysm that was found on her grandmother's autopsy. Based on pregnancy planning and the current literature, endovascular management with a covered stent was proposed. CASE REPORT: Self-expandable, covered stent (Bard, Fluency®) was implanted using a single transfemoral approach. A stiff guidewire and a large sheath distorted the anatomy, which resulted in an incomplete aneurysmal neck covering. In the absence of additional covered stents, the procedure was terminated. Two weeks later, computed tomographic angiography (CTA) confirmed persistent aneurysmal perfusion due to the incomplete neck coverage. A multidisciplinary board opted for a second endovascular attempt, this time with a longer covered stent via the transaxillary approach in order to reduce anatomical distortion. Balloon, expandable, cobalt-chrome covered stent (Jotec, E-ventus BX®) was implanted in the SMA, covering the aneurysmal neck and overlapping the previously implanted covered stent. Angiography confirmed a complete exclusion of the aneurysm. A control US performed three weeks later confirmed a patent covered stent and complete aneurysmal exclusion. There was a mild median nerve damage periprocedurally that resolved in three months. The most recent US control examination, performed eleven months after the procedure, showed an excluded aneurysm and a patent covered stent. There were no clinical signs of bowel ischaemia during the follow-up period. CONCLUSIONS: Endovascular management of SMAA proved to be safe and efficient. The "access from above" is probably safer and should be considered in the majority of cases with acceptable sizes of access vessels. Mid-term results in our patient are good and life-long follow-up is planned to prevent late complications.

Successful Treatment of Acute Aortic Dissection Type Stanford A Presenting as Limb Ischemia, Successfully Treated with Operative and Endovascular Procedures.

We report a successful treatment of unusual case of a 48 year old male patient with acute aortic dissection type Stanford A that expanded into left common and external iliac artery diagnosed by MSCT angiography, presenting as a single leg paresis, without symptoms of a chest or back pain. Patient was operated with conventional ascending aortic replacement. Patient had no known prior medical condition. He has been treated for acute thrombosis of the left popliteal artery developing one day after ascending aortic replacement surgery, embolectomy was performed. Critical limb ischemia developed due to preocclusive stenosis of the left common and left external iliac artery and was treated by endovascular procedure of iliac artery stenting performed on the fifth postoperative day. After 17 days patient was discharged form hospital, showing no neurological or vascular deficit. For successful treatment of acute aortic dissection type Stanford A complicated with limb ischemia, rapid and accurate diagnosis is essential, together with close cooperation of cardio surgeons, vascular surgeons and invasive radiologists and individual approach to these demanding patients.

Predicting carotid restenosis by comparison of plaque MCP-1 mRNA expression and serum levels.

As cardiovascular pathology grows in numbers, research into the discovery of new chemokine biomarkers should not be neglected, as they seem to be paramount in atherosclerosis prevention and its early detection. Chemokines attract and activate leukocytes and are well recognized in the environment of inflammatory response. MCP-1 is a valuable chemokine whose potential to become a new crucial atherosclerosis marker is surely worth investigating. Since quantities of MCP-1 found in lesions are as low as immeasurable, we propose the use of an immunohistochemical method for the quantification of MCP-1 levels in atherosclerotic lesions. Additionally, serum levels of MCP-1 can be measured by commercially available immunoassays. Proposed MCP-1 concentration increase may explain the acceleration in lesion's atherosclerosis progression as chemokine activation occurs once they bind to specific ligands. If proven, this hypothesis would indicate the need for further studies in order to objectively link the increased MCP-1 expression with carotid restenosis.

Significance of mini radial endoscope ultrasound (MREUS) in diagnosis and therapy of colorectal neoplasia.

In the pursuit to advance diagnostic procedures with colon carcinoma patients, we included the 15 MHz mini radial endoscopic ultrasound (MREUS) in our work up algorithm, following PH verification. When compared to surgical and final pathohistological (PH) findings, MREUS shows that it can differentiate colon layers in great detail and therefore we can determine the degree of carcinoma dissemination (T1, T2, T3, T4) as well as to make a correct therapeutic choice. MREUS (12-15-20 MHz) is a highly reliable colon layer structural analysis method. As we have shown in our study, the accuracy of T stage colon carcinoma visualization in correlation to equivalent PH studies varies from 90-100% which makes MREUS the best as well as the most reliable method in determining preoperative T stage colon carcinoma.

Drug usage by outpatients in Croatia during an 8-year period: Influence of changes in pricing policy.

OBJECTIVE: The aim of our study was to investigate the changes in drug usage and financial expenditure according to legal changes in Croatia during the period 2001 - 2008, especially considering pricing policy. MATERIALS: The data on outpatient drug usage during the studied period was obtained from the Croatian National Health Insurance (CNHI). CNHI maintains a database on drugs prescribed by primary health care physicians and dispensed by pharmacies. METHODS: The data was calculated and presented in defined daily doses (DDD) per inhabitant per year for antibiotics and in DDD/1,000 inhabitants/day for other drugs. The data is also presented in Euro/DDD and the financial expenditures are presented in Euros. RESULT: During the investigated period drug usage increased 81.33%, while financial expenditure increased 77.23%. While total DDD/1,000 increased ~ 10% every year, financial expenditure increased 10 - 20% annually until 2006, but since then there have been no significant changes. CONCLUSION: Pricing policy changes could influence drug financial expenditure considerably in the short-term, but it is also important to apply a combination of measures for drug expenditure control. Numerous interventions from authorities from different countries all over the world, prove that there is still no so called "gold standard" which could restrain growing usage and expenditure of drugs. Clinical pharmacologists and clinical pharmacists should be included in these processes.

Vomiting as a reliable sign of concussion.

Concussion is the most common type of traumatic brain injury, with headache being the most frequent symptom of mild traumatic brain injury (MTBI) (including dizziness, vomiting, nausea, lack of motor coordination or difficulty balancing). Concussion may be caused by a blow to the head, or by acceleration forces without a direct impact. Often, MTBI occurs as the result of a sports injury. Loss of consciousness is always present, unlike vomiting. Therefore, we hypothesize vomiting to be considered as a cardinal sign of concussion. Stimulation of vomiting centres finally triggers vomiting. Professional boxers and mixed martial arts competitions reluctantly agree with stringent rules and protective clothing. We discuss the issue of further protection for those engaged in these and other sports.

Advantages of intramedullar fixation in treatment of congenital tibial pseudoarthrosis--a case report.

This report describes a case of a 29-year old patient with congenital pseudoarthrosis of the distal tibia previously treated unsuccessfully by a conventional surgical method. Tibial congenital pseudoarthrosis is a rare disease characterized by segmental osseous weakness resulting in deformation of the bone and spontaneous fractures which progresses to a tibial nonunion. In our case we used intramedullary stabilization with bone grafting and six month after operation congenital pseudarthrosis of the tibia healed.

Functional outcome assessment of lower limb amputees and prosthetic users with a 2-minute walk test.

The aim of this study was to assess the functional outcome of a population of lower limb amputees supplied with prosthesis. The research was conducted from June to September of 2010 at the Center for Physical and Rehabilitation Medicine, of the Clinical Hospital Center Rijeka, Croatia. The study included 50 adult subjects of both genders with a unilateral transtibial or transfemoral lower limb amputations. The 2-minute walk test (2MWT) was used to assess the functional outcome of these individuals. Data were statistically analyzed. Subjects were divided into groups according to age. The best results were obtained by subjects between the age of 45 and 59 years. The difference between groups was statistically significant (p < 0.001). Taking into account the cause of amputation, there was a statistically significant difference in the results of the 2 MWT between subjects in whom the cause of amputation was circulatory and those where the cause of the amputation was not due to circulatory problems. The best results were obtained in subjects in whom the cause of amputation was not circulatory (p = 0.009). Considering the level of amputation there was a statistically significant difference in the results of the 2MWT between subjects with transtibial and those with transfemoral amputations. Better results were obtained in transtibial amputees (p = 0.039). Considering the first prosthetic supply, better results were obtained in subjects using prosthetic devices over 9 years (p = 0.031). Our research confirmed that age, gender, level and cause of amputation, including the time from the first prosthetic supply have an effect on the 2MWT results.

Early identification of patients with the risk for postoperative carotid restenosis development.

Multiple randomized trials over the last decade for both symptomatic and asymptomatic carotid stenosis have proven the efficacy of carotid endarterectomy (CEA) in reducing the risk of stroke. The aim of this prospective non-randomizing cohort study was to determine the incidence of carotid arteries restenosis after CEA as well as to ascertain the clinical and etiological characteristics for the development of restenosis. Treatment data from 178 KBC Rijeka patients that had undergone CEA in the period 1. 09. 2005-30. 8. 2009 has been processed. All patients are monitored trough our Neurosonology laboratory algorythm--first Doppler ultrasound examination within the first week after CEA and the following after 1, 3, 6 and 12 months. After this time once a years. The average monitoring time was 21 month (1-36 months). In the stated period 27 restenosis was diagnosed (15.16%). Only four of them were symptomatic (14.81%). Patient survival rate is 98% in the first 12 and 92% in the first 36 months. Carotid restenosis is usually asymptomatic. Non-invasive postoperative carotid arteries color Doppler screening is essential in the early identification of patients with the risk for the development of restenosis.

Comparison between carotid stenting and carotid endarterectomy in early outcome.

Carotid artery stenting (CAS) is a widely used method in prevention of stroke for carotid artery stenosis as an alternative to surgical treatment. Initial studies reveal higher morbidity and mortality rates for CAS than acceptable standards for carotid endarterectomy (CEA). The aim of this study was to compare results in a series of CAS with concurrent risk-matched group of CEA patients. The study included two groups of 50 patients with internal carotid artery stenosis. We compared early outcome (30 days after procedure) in risk-matched groups of patients that underwent these procedures. Post procedural complications were equally frequent in both groups. There was no significant difference in perioperative complication rates (P = 0.871). Comparison of these two methods shows that CAS and CEA are competitive methods for treatment of carotid artery stenosis. Particularly in symptomatic patients with high risk for surgery CAS is alternative treatment.

The transition from pulpitis to periapical periodontitis in dogs' teeth.

The purpose of the study was to histologically analyse transition from pulpitis to periapical periodontitis on dog's teeth. Pulps of mandibular premolars (37 roots) were exposed using a low-speed handpiece. Teeth were left open to the oral environment for 20, 35, 50 and 65 days. After the experimental period animals were sacrificed. Undemineralised teeth with surrounding bone, embedded in methylmetacrylate, were prepared for standard histological analysis. All teeth with pulpitis (five roots), regardless of the experimental period, had acute serose periapical periodontitis. All teeth (15 roots) with partial pulp necrosis had subacute periapical periodontitis. Teeth with complete pulp necrosis (19 roots) had chronic periapical periodontitis and in one case suppurative apical periodontitis. The condition of the pulp correlates with the histopathological findings of periapical tissue in the open types of pulp infection. Acute periapical periodontitis begins during pulpitis and can occur before 20 days of pulp exposure in the dog.

Apoptosis and cardiopulmonary bypass.

AIM: The aim of this study was to ascertain the percentage of apoptotic myocytes in patients who underwent coronary artery bypass surgery. Apoptotic index (AI) obtained with in situ terminal deoxynucleotidyl transferase-labeled dUTP nick end labeling (TUNEL) method and Bak protein expression were compared. PATIENTS AND METHODS: Twenty consecutive patients who underwent coronary artery bypass surgery, myocardial samples from the right atrium were taken in three stages: before cannulation (the first sample group), after declamping (the second sample group), and 20 minutes after reperfusion (the third sample group). The percentage of apoptotic cells was determined by TUNEL method. Expression of Bak protein was immunohistochemically analyzed. Intermittent ischemia and moderate hypothermia were used as methods of myocardial management during surgery. A statistical analysis was performed by using the Friedman ANOVA analysis of variances, the Kendall coefficient of concordance and the Wilcoxon matched pair test. RESULTS: In the first sample group mean value of Bak expression was 2.61 +/- 2.18, compared with AI 5.38 +/- 3.58, after declamping (the second sample group) the mean value of Bak expression was 4.31 +/- 2.68 while AI was 7.63 +/- 4.38 and after 20 minutes of reperfusion in the third sample group mean value of Bak expression was 8.89 +/- 4.45, while AI was 15.6 +/- 8.45. When compared by using Wilcoxon matched pair test two methods significantly correlated, p > 0.0001. CONCLUSION: The positive correlation between AI obtained by TUNEL method and expression of Bak protein may suggest that apoptosis is activated mainly through mitochondrial activation pathway in ischemic reperfusion injury. The results suggest that ischemic reperfusion injury increases the AI in the right atrial tissue. If so, immunohistochemical expression of Bak protein could be used as a marker of myocardial ischemia induced injury.

Ochronosis of the aortic valve and aorta.

A 64-year-old male patient with unknown alkaptonuria and severe aortic stenosis and ischemic heart disease was admitted to the authors' institution for elective surgery. The patient underwent aortic valve replacement with a 25-mm aortic valve (ATS Medical, Inc.) and single venous aortocoronary artery bypass grafting for a right coronary artery. Aortotomy revealed typical ochronotic pigmentation of a severely calcified aortic valve and aortic intima. A diagnosis of alkaptonuria was confirmed by evidence of homogentisic acid in the patient's urine, together with histopathological analysis.

Chronic heart perforation with 13.5 cm long Kirschner wire without pericardial tamponade: an unusual sequelae after shoulder fracture.

We report a unique case of cardiac embolization with the Kirschner wire that has been used for osteosynthesis for 24 months previously. According to the complete analysis of medical records and autopsy report, the wire had migrated from the right humeroscapular joint to the heart. Although migration of a Kirschner wire has been reported in the literature, migration of the wire with a total length of 13.5 cm with no pericardial tamponade, despite myocardial perforation, has not been previously described.

Pulmonary valve papillary fibroelastoma. A case report.

We present a case of pulmonary valve fibroelastoma diagnosed by echocardiogram which was confirmed by surgical resection in a patient in whom elective coronary artery bypass surgery was performed. The patient had no clinical or constitutional symptoms suggesting the presence of an intracardiac tumor. Routine preoperative transthoracic echocardiography revealed a pulmonary valve tumor. Histopathological analysis resulted in a diagnosis of papillary fibroelastoma. Papillary fibroelastomas are rare and benign cardiac tumors. They usually arise from the cardiac valves. In the literature we found only a few cases, so it seems there is a need for further description of additional cases of pulmonary valve fibroelastoma.