Setting accelerated dissolution test for PLGA microspheres containing peptide, investigation of critical parameters affecting drug release rate and mechanism.

The objective of this study was development of accelerated in vitro release method for peptide loaded PLGA microspheres using flow-through apparatus and assessment of the effect of dissolution parameters (pH, temperature, medium composition) on drug release rate and mechanism. Accelerated release conditions were set as pH 2 and 45°C, in phosphate buffer saline (PBS) 0.02M. When the pH was changed from 2 to 4, diffusion controlled phases (burst and lag) were not affected, while release rate during erosion phase decreased two-fold due to slower ester bonds hydrolyses. Decreasing temperature from 45°C to 40°C, release rate showed three-fold deceleration without significant change in release mechanism. Effect of medium composition on drug release was tested in PBS 0.01M (200 mOsm/kg) and PBS 0.01M with glucose (380 mOsm/kg). Buffer concentration significantly affected drug release rate and mechanism due to the change in osmotic pressure, while ionic strength did not have any effect on peptide release. Furthermore, dialysis sac and sample-and-separate techniques were used, in order to evaluate significance of dissolution technique choice on the release process. After fitting obtained data to different mathematical models, flow-through method was confirmed as the most appropriate for accelerated in vitro dissolution testing for a given formulation.

Brain and art: illustrations of the cerebral convolutions. A review.

BACKGROUND: Aesthetics and functional significance of the cerebral cortical relief gave us the idea to find out how often the convolutions are presented in fine art, and in which techniques, conceptual meaning and pathophysiological aspect. MATERIALS AND METHODS: We examined 27,614 art works created by 2,856 authors and presented in art literature, and in Google images search. RESULTS: The cerebral gyri were shown in 0.85% of the art works created by 2.35% of the authors. The concept of the brain was first mentioned in ancient Egypt some 3,700 years ago. The first artistic drawing of the convolutions was made by Leonardo da Vinci, and the first colour picture by an unknown Italian author. Rembrandt van Rijn was the first to paint the gyri. Dozens of modern authors, who are professional artists, medical experts or designers, presented the cerebralc onvolutions in drawings, paintings, digital works or sculptures, with various aesthetic, symbolic and metaphorical connotation. Some artistic compositions and natural forms show a gyral pattern. The convolutions, whose cortical layers enable the cognitive functions, can be affected by various disorders. Some artists suffered from those disorders, and some others presented them in their artworks. CONCLUSIONS: The cerebral convolutions or gyri, thanks to their extensive cortical mantle, are the specific morphological basis for the human mind, but also the structures with their own aesthetics. Contemporary authors relatively often depictor model the cerebral convolutions, either from the aesthetic or conceptual aspect. In this way, they make a connection between the neuroscience and fineart.

Heart in anatomy history, radiology, anthropology and art.

BACKGROUND: Anthropologic, artistic and medical significance of heart inspired usto undertake this multidisciplinary study. MATERIALS AND METHODS: Amongst the 24 obtained echocardiograms and phonograms, 1 was used for a Photoshop processing. In addition, over 20,000 art work reproductions were examined in this study. RESULTS: Artistic and symbolic presentation of heart started some 15,000 years ago. First heart models were made by the Egyptian and Olmec civilisations. Ancient cultures regarded heart as the seat of the soul, spirit and intelligence. First anatomical and artistic images of heart were created by Leonardo da Vinci in the15th century, and first wax models by the Italian anatomists in the 17th century. Mediaeval religious symbolism of heart was replaced in the Renaissance and later on mainly by its role in the romantic love. Anatomical heart art continued in the 18th and 19th centuries through the works of Sénac, Cloquet, Hirschfeldand Bourgery. Some modern artists, such as Dalí, Kahlo, Rivera, Warhol, Ivanjicki, Vital, Kober and Mastrlova, created the anatomical heart images or sculptures, whereas some others, such as Duchamp, Klee, Miró, Matisse and Dine, presented heart symbol in their artworks. New radiologic technologies produce fine images of heart, some of which are similar to the works of modern artists. CONCLUSIONS: Heart biology and symbolism have had a tremendous influence on our culture, including art and medical sciences. New radiologic techniques and computer technology have produced such images of heart, which substantially improved diagnosis, but also enhanced the heart aesthetics.

Repair of abdominal aortic aneurysms in the presence of the horseshoe kidney.

AIM: Horseshoe kidney is the most common congenital kidney anomaly, occurring in 0.15-0.25% of all newborns. A medial fusion of the kidneys, mostly anteriorly to the aorta, is the main characteristic of this anomaly. The co-existence of abdominal aortic aneurysm (AAA) and horseshoe kidney is rare, occurring only in 0.12% of patients. The aim of this paper is to define the optimal management of patients with AAA associated with the horseshoe kidney. METHODS: This paper presents the analysis of patients operated at the Clinic for Vascular and Endovascular Surgery of the Clinical Center of Serbia in Belgrade due to AAA associated with the horseshoe kidney as well as the analysis of the previously published literature data regarding this topic. RESULTS: Between 1985 and 2011, data were collected retrospectively on 25 patients with the horseshoe kidney who underwent aortic surgery. Out of them, 6 patients had aortoiliac occlusive disease and 19 patients had aortic aneurysm. More detailed analysis of the aneurysmatic group was performed. Among them there were 16 male and three female patients, with the average age of 63.8 (50-76) years. Two patients had type IV of thoracoabdomial aortic aneurysm (TAA) according to Crawford-Saffi classification, while 17 had infrarenal abdominal aortic aneurysms. There were 15 elective and four urgent procedures due to aneurismal rupture. The presence of the horseshoe kidney was detected in 16 patients before surgery (84.2%) by means of Duplex ultrasonography, angiography, computed tomography and intravenous urography. Multiple renal arteries were presented in 12 (63.2%) cases. A transperitoneal approach was used in 16 cases with abdominal aortic aneurysm, while left retroperitoneal approach with partial extrapleural removal of the 11th rib was performed in two cases of thoracoabdominal aneurysm and in one patient with AAA. In 18 cases, kidney tissue transection was successfully avoided with vascular graft placement beneath the horseshoe kidney. In one case only, the division of the renal isthmus was performed. In all 12 cases with detected anomalous renal arteries, their reattachment into vascular graft has been performed. Two patients (10.5%) died during perioperative period. One of them had ruptured type IV TAA. Seventeen patients who survived were followed from one to twenty years (mean 6.6 years). During the follow up period we lost track of 4 patients. In this period there were no signs of graft occlusion, or renal failure. CONCLUSION: Repair of an abdominal aortic aneurysm in the presence of the horseshoe kidney is a truly particular surgical challenge. It is associated with three main problems: choice of the surgical approach; the procedure regarding kidney isthmus preservation as well as recognition and reattachment of all significant anomalous renal arteries.

Neuron specific enolase tissue expression as a prognostic factor in advanced non small cell lung cancer.

PURPOSE: To determine the frequency of neuron specific enolase (NSE) tissue expression and its possible influence on survival of patients treated for advanced non small cell lung cancer (NSCLC). PATIENTS AND METHODS: We studied 158 patients with histological diagnosis of NSCLC (stage III/IV). Combined chemoradiotherapy was used in stage III disease (without pleural effusion), whereas chemotherapy only was used in stage III (with pleural effusion) as well as in stage IV disease. Immunohistochemical analysis of NSE expression was determined using antibodies to NSE (DAKO, Den). One- and 2-year overall survival were assessed. RESULTS: 45 (28.5%) patients had positive NSE expression. The most frequent NSE expression was seen in 6/9 (66.7%) patients with large cell carcinoma and in 23/50 (46%) with adenocarcinoma. One- and 2-year survival rates were 70% and 30% in the group of patients with positive NSE expression compared to 26% and 3% in the group with negative NSE expression (p=0.000). The median survival time was 16.4 vs. 11.4 months in the groups of patients with and without NSE expression, respectively (p < 0.001). One-and 2-year survival rate was higher in NSCLC patients with more than 50% of NSE positive cancer cells (p=0.0004 and 0.000, respectively). CONCLUSION: A total of 28.5% of advanced NSCLC patients had positive NSE expression. Median 1- and 2-year survival time was significantly longer in patients with positive NSE expression.

Construction and characterization of a recombinant plasminogen activator composed of an anti-fibrin single-chain antibody and low-molecular-weight urokinase.

BACKGROUND: Targeting of plasminogen activators to the fibrin component of a thrombus by antibodies directed against human fibrin can enhance their thrombolytic potency and clot specificity. OBJECTIVES: To overcome the disadvantages of chemical conjugation, we investigated whether the recombinant fusion of a single-chain antibody and a plasminogen activator results in an active bifunctional molecule that might be useful as a therapeutic agent. METHODS: The cDNA of low-molecular-weight single-chain urokinase-type plasminogen activator, comprising amino acids Leu144-Leu411 (scuPA(LMW)), was cloned from human endothelial cells and fused to a single-chain antibody specific for the 7 N-terminal amino acids (beta(15-22)) in the beta-chain of human fibrin (scFv(59D8)). The fusion protein was purified using affinity chromatography with the beta(15-22)-peptide of human fibrin. RESULTS: Purified scFv(59D8)-scuPA(LMW) migrated as a 60-kDa band, which is consistent with a molecule composed of one scFv(59D8) and one scuPA(LMW) moiety. Both functions of the fusion molecule, fibrin-specific binding and plasminogen activation, were fully preserved. In human plasma clots, thrombolysis by scFv(59D8)-scuPA(LMW) is significantly faster and more potent compared with the clinically used urokinase. CONCLUSIONS: ScFv(59D8)-scuPA(LMW) constitutes a new recombinant chimeric plasminogen activator with a significantly enhanced thrombolytic potency and relative fibrin selectivity, that can be produced with modern methods at low cost, large quantities and reproducible activity in Escherichia coli.

[Symptoms of locally advanced non-small-cell lung carcinoma]. / Simptomi lokalno uznapredovalog nesitnocelijskog karcinoma pluca.

A group of 50 patients with locally disseminated non-small-cell lung cancer was analyzed. The diagnosis was confirmed upon the analysis of the samples obtained by bronchoscopy or percutaneous needle biopsy of the lungs. CT scan of the thorax, as well as transesophageal echosonography of the mediastinum and ultrasound of the abdomen were performed for the evaluation of the disease dissemination. The aim of the study was to establish the main characteristics of lung cancer in locally disseminated disease with particular reference to the analysis of the symptoms. The majority of patients were aged between 56 and 60 years (n = 13; 26%). The majority was with planocellular cancer of central localization (n = 38; 76%). In 20 (40%) patients surgery was performed, and in 30 (60%) patients the surgery was finished with exploration. The most frequent symptoms were cough in 29 (58%), hemoptysis in 27 (54%) and chest pain in 24 (48%) patients. Frequency of the symptoms was multiplied in the patients with the advanced disease. Frequency of hemoptysis was equal in the group of patients who were operated and those with exploratory thoracotomy.

Hyperbaric oxygenation as adjuvant therapy to surgery of emphysematous cholecystitis.

Three cases of emphysematous cholecystitis are presented. The role of hyperbaric oxygenation as excellent adjuvant therapy to urgent surgical as well as intensive conservative treatment is emphasized.

[Pulsed doses of cyclophosphamide in the treatment of Wegener's granulomatosis]. / Pulsne doze ciklofosfamida u lecenju Vegenerove granulomatoze.

The aim of the study was to analyze the efficacy and toxicity of the therapy with pulse dose of cyclophosphamide in patients with Wegener's granulomatosis. The diagnosis was established in three patients (2 male, 1 female, average age 58.3 years) upon the clinical picture, laboratory-immunologic and histopathologic findings. The initial therapy was conducted by pulse doses of cyclophosphamide (0.6 and 0.8 g/kg), glucocorticoids (0.5 g), repeated plasmapheresis and pulse doses of immunoglobulin (0.4 g/kg). All patients achieved remission with complete recovery of renal function in one patient. The therapy with pulse doses of cyclophosphamide (time-period between pulses from 1 to 3 months), together with glucocorticoids taken orally for 24 months, was continued for the following 18 months. Stable remission (4-15 months) was maintained in all the patients after the immunosuppressive therapy was over. The treatment of Wegener's granulomatosis by cyclophosphamide and glucocorticoid pulse doses with the application of plasmaphereses and immunoglobulin pulse doses in the initial therapy lead to the remission of progressive types of long duration.

[Endobronchial metastases in extrapulmonary malignant tumors]. / Endobronhijalne metastaze vanplucnih malignih tumora.

Endobronchial metastases of extrapulmonary malignant tumors are very rare and they appear in 1-2% of all the pulmonary metastases cases. By their endoscopic presentation, they are similar to primary bronchogenic carcinoma. Eleven patients with endobronchial metastases were presented, 8 male and 3 female. The average age was 58.27 years (from 37 to 72 years). Primary renal carcinoma existed in 2 patients. The one was operated 5 years before the metastases appeared in bronchus, and the metastases in bronchial wall contributed to the discovery of primary tumor in the other patient. Endobronchial metastases appeared in 3 patients, 2 years after the surgery of malignant colonic or rectal tumor, and in fourth patient it appeared 5 years after the surgery. One female patient sustained mastectomy and the radiation therapy was performed 11 years before the bronchial metastases occurred and the other female patient had the disease revealed a year ago and the polytherapy was performed. In the female patient with non Hodgkin Lymphoma, the change in bronchus was revealed simultaneously as the primary disease. Endobronchial metastases appeared in 2 patients two years after the surgery of malignant melanoma. The disease diagnosis was set by clinical-radiological examination, by endoscopy and by the comparison of histopathologic finding of primary carcinoma and metastases.

[Personal experience in the treatment of tuberculosis in patients in war areas]. / Nasa iskustva u lecenju tuberkuloze kod obolelih sa ratnih podrucja.

In the four-year period (1991-1995), 96 tuberculosis patients from the war areas of former Yugoslavia were treated in the Clinic for Lung Diseases of Military Medical Academy, that makes 31% of total number of sick and treated for tuberculosis-309 (100%). In group I (patients from war areas) there were 45 cases of cavernous pulmonary tuberculosis (47%), 28 bilateral (29%), 16 pleurisy (16.7%) and 6 cases of extensive pulmonary tuberculosis (5.1%). Among 213 patients (100%) from group II (patients from FR Yugoslavia), 81 patients had cavernous pulmonary tuberculosis (38%), 49 patients had bilateral (23%), 33 patients had pleurisy (15.5%) and 6 patients had extensive pulmonary tuberculosis (2.8%). In group I the diagnosis was bacteriologically and/or histopathologically proved in 94.8% cases: by finding of acid-resistant bacilli in sputum and other biological materials in 69 patients (72%), by positive Löwenstein's cultures in 73 patients (76%) and by histopathological result of tuberculosis inflammation in 34 patients (35.4%). In group II tuberculosis was bacteriologically and/or histopathologically proved in 134 patients (63%) by acid-resistant bacilli in sputum and other biological materials, in 141 patient (66%) by positive Löwenstein's culture and in 71 patient by positive histopathological results. Pulmonary tuberculosis in war areas is characterized by greater frequency of severe clinical forms (cavernous, bilateral and extensive) and by high direct bacillarity.

The arachidonic acid cascade, eicosanoids, and signal transduction.

Eicosanoid biosynthesis in animal cells either results from agonist-stimulated phospholipase activation (endogenous pathway) or from lipoprotein receptor-mediated uptake and lysosomal lipid hydrolase-dependent release of AA (exogenous pathway) (see Fig. 1 for schematic representation). LDL stimulates eicosanoid formation through delivery of substrate AA to enzymes of oxidative AA metabolism. The classical LDL receptor is a control point of the effects of LDL AA on eicosanoid formation in different tissues: LDL AA metabolism occurs in several cell types of mesenchymal and epithelial origin and generates the formation of distinct eicosanoid patterns in each case. The LDL AA pathway does appear to couple directly to the PGH synthase reaction, whereas it does not couple directly to the 5-lipoxygenase reaction. We expect that a more complete characterization of the LDL unsaturated fatty acid pathway in different tissue will yield additional information on the biochemistry of lipoproteins, AA, and eicosanoids.

[A rare case of secondary carcinoma of the colon]. / Rijedak slucaj sekundarnog karcinoma kolona.

Primary gelatinous tumors of colon are very rare. There is a case of metastatic colon's tumor as a result of transcaelomic expansion of gelatinous cancer of stomach, there is a possibility of implantation tumor's cells during appendectomy.

[The obstruction syndrome in patients with bronchogenic carcinoma]. / Opstrukcijski sindrom u bolesnika sa bronhogenim karcinomom.

In the period 1985-1990, 760 patients with bronchogenic carcinoma have been examined and treated at the Clinic of Lung Diseases of the Military Medical Academy. Obstruction syndrome was found in 64% of patients of whom 21% with severe obstruction. Pulmonary insufficiency, most commonly of milder, partial type, was found in 19% of patients. Due to the limited lung function as a consequence of chronic obstructive lung disease (COLD) 45 patients with locally demarcated tumours were not operated, surgical resection was not possible. In the operated patients with obstruction syndrome the risk of postoperative complications was increased. COLD and limited lung function most commonly had not been the obstacle for application of radiotherapy of bronchogenic carcinoma.

[Alveolitis in patients with sarcoidosis]. / Alveolitis kod bolesnika od sarkoidoze.

In 20 patients with pulmonary sarcoidosis bronchoalveolar lavage (BAL) was performed. The cellular content was analysed and lymphocyte subpopulation was determined using monoclonal antibodies. The patients with more than 28% of BAL T lymphocytes are classified as patients with high intensity alveolitis (n = 8) and those with less than 28% of T lymphocytes as low intensity alveolitis (n = 12). Of the total number of BAL T lymphocytes in high intensity alveolitis CD 3+ T cells were 88.8 +/- 8.4% and in low intensity alveolitis 68.2 +/- 13.8% (p < 0.001). In high intensity alveolitis the number of CD 4+ T cells was increased and the number of CD 8+ T cells was decreased which caused the increased index CD 4:CD 8; 9.8 +/- 8.7. In low intensity alveolitis this ratio was 2.2 +/- 1.2. Effects of smoking were also analysed. In smokers the total number of BAL cells was higher 184.0 +/- 47.2 x 10(4) ml of BAL, and in nonsmokers 101.3 +/- 65.1 x 10(4) ml of BAL (p < 0.001). Smoking has no effects on the characteristics of the cellular elements and subpopulation of BAL lymphocytes in patients with sarcoidosis. In 7 patients T lymphocyte subpopulation in the peripheral blood were determined too. Patients with high intensity alveolitis had the increased index CD 4:CD 8 in the peripheral blood compared with low intensity alveolitis.

[Personal experience in the diagnosis and treatment of pulmonary embolism]. / Nasa iskustva u dijagnostici i lecenju plucnog embolizma.

The 188 patients with pulmonary embolism were treated at the Clinic for Pulmonary Diseases of the M.M.A. in the period 1979-1989. Clinical symptoms and signs, diagnostic procedures and therapy of pulmonary embolism are analysed. The diagnosis is difficult and is suggested by the predisposing factors, symptoms and signs of the disease as well as indirect diagnostic procedures. The contribution of perfusion lung scintigraphy is precious. Deep venous thrombosis of the lower extremities, the source of pulmonary embolism, is commonly clinically absent. The complete cure without complications was achieved in 156 (83%) patients. The most common complications were: early and late recurrences, hematuria and peptic bleeding. The lethal outcome occurred in two cases of basic disseminated malignant disease.

[Viscerosynthesis--use of mechanical sutures in digestive tract surgery]. / Viscerosinteza--primjena mehanickog sava u digestivnoj kirurgiji.

Mechanical suture (stapler) was applied 164 times in digestive surgery in the period from October 1985 to December 1988. There have been performed 82 anastomoses (30 low collateral, 20 esophagointestinal, 20 ileocolic, 10 enteroenteral and 2 colocolic) and 76 closure of intestines. Anastomoses were performed with EEA stapler in 50, GIA stapler in 30 and TA stapler in 2 cases. Intraoperative complications related to the use of stapler were observed in 4.8% cases. Clinically manifested dehiscence of the stapler anastomoses was evident in 4.8% (82) and immediate cause of death in 1.2% (1) case. The incidence of secondary wound infection after four years was 2.4%. No clinically manifested stenosis of anastomoses were observed within the follow up period from one to four years.

[Analysis of immunocompetent cells in bronchoalveolar lavage in patients with sarcoidosis]. / Analiza imunokompetentnih celija u bronhoalveolnom lavatu bolesnika sa sarkoidozom.

Bronchoalveolar lavage through fiberoptic bronchoscope was performed in the right middle lobe or lingula of 8 patients with clinically and radiologically active sarcoidosis and 5 healthy persons. Pellet cells were analysed. Lymphocyte subsets were detected with monoclonal antibodies by fluorescent microscopy or flow cytometry (EPICS-C, Coulter). In patients with sarcoidosis lymphocyte proportion is elevated (29.88 +/- 4.32) as compared to healthy persons (5.2 +/- 3.4). We detected an increase of CD 3 lymphocytes (86.38 +/- 6.71) and CD 4 helper-inducer T lymphocytes compared to the total number of lymphocytes and, likewise, of the CD 4/CD 8 ratio--5.62 +/- 0.94. CD 8 suppressor-cytotoxic T lymphocytes are decreased (13.0 +/- 1.3 as compared to the healthy persons 22.0 +/- 3.84), which is a relative decrease because due to an increase of the total lymphocyte number, an increase of CD 8 T lymphocytes (3.85 +/- 0.41 as compared to the healthy persons 1.09 +/- 0.59) still exists, compared to the total number of pellet cells.