Polyphenol rich extracts of Geranium L. species as potential natural antioxidant and antimicrobial agents.

OBJECTIVE: Plants and plant extracts are of great scientific interest due to the chemical diversity and pharmacological properties of present bioactive molecules. The Geranium L. species are widely used in ethnomedicine. In the current study, the total phenolic and tannin content, antioxidant and antimicrobial activity of methanol extracts of eight Geranium species were investigated. MATERIALS AND METHODS: The total phenolic and tannin content were determined by the FC method. Antioxidant capacity was evaluated in FRAP, DPPH, and biochemical assays, while antimicrobial activity was examined using the broth microdilution method. RESULTS: The high total phenolic (170.64-636.32 mg GAE/g dry extract) and tannin content (37.80-414.02 mg GAE/g DE), along with significant total antioxidant (FRAP values 1.13-8.80 mmol Fe2+/g) and DPPH radical scavenging activity (SC50 values 4.24-34.52 µg/mL) were observed. The prominent antioxidant capacity was confirmed in biochemical assays (OS values -1.47 - -13.02). The extracts exhibited significant antimicrobial activity against ATTC strains (MICs dominantly in the range of 12.5-200 µg/mL) as well as against clinical isolates of E. coli (MICs mostly 50 and 100 µg/mL). The pronounced antioxidant and antimicrobial activity can be due to the high phenolic content, particularly due to the presence of hydrolyzable tannins. CONCLUSIONS: Based on the high content of polyphenols, pronounced antioxidant and antimicrobial activities, the examined extracts are promising natural antioxidant and antimicrobial agents with the potential medicinal purpose and use as a functional food.

Integrating patient safety education into early medical education utilizing cadaver, sponges, and an inter-professional team.

BACKGROUND: Introducing patient safety and quality improvement science to medical students is integral to improving healthcare. However, developing and implementing a patient safety curriculum can be challenging in a medical school curriculum that is already densely packed. Our aim was to develop and evaluate the impact of a workshop introducing patient safety and quality improvement science to a large class of first-year medical students. METHOD: As a part of an evolving longitudinal patient safety curriculum, an introductory workshop on patient safety was integrated into an anatomy course. A high impact event (a simulated "retained sponge" discovery during an anatomy dissection lab) was used to introduce medical error. The educational session which followed consisted of a presentation by an interprofessional team utilizing the retained sponge as example of an error. Use of safety tools was introduced and quality improvement science was discussed using the evolution of methods to decrease retained foreign objects during surgery. A patient's story told by a close family member about the personal impact of medical errors was presented. Students then participated in an interactive breakout activity and completed a module on safety. The impact of the workshop was assessed through pre- and post- session tests. RESULTS: Quantitative and qualitative evaluation reflected a positive effect of the session in improving students' safety knowledge and attitudes. Students' mean total knowledge improved from 7.58 to 8.98 (p = 0.000). Mean total attitudes score improved from 47.73 to 50.56 (p = 0.000). Students' comments after the workshop reflected increased awareness and appreciation of the importance of addressing medical errors. CONCLUSION: A workshop introducing patient safety and quality improvement to first year medical students improved knowledge and attitudes regarding safety and increased awareness of the importance of addressing medical errors in their future careers. Integrating patient safety education into an existing foundational science course is a model for teaching patient safety at other medical schools.

Tolerability and toxicity of lipidosterolic extract of American dwarf palm Serenoa repens in Wistar rats: well-known extract, new insight.

BACKGROUND AND OBJECTIVES: Serenoa repens extracts (SrE) have been used for centuries in the treatment of benign prostatic hyperplasia (BPH). According to recommendations that each product should be examined separately, including its tolerability and toxicity, we conducted this study in order to broaden the current cognition about tolerability and toxicity of SrE, in particular of German brand ProstamolunoR. MATERIALS AND METHODS: Twenty-four adult male Wistar rats were randomly distributed into 4 groups of 6 animals. The first control group (O) received water (1 ml/kgBW) and second control group (OO) received olive oil (1 ml/kgb.w.) every day for 30 days. The third and fourth group of rats (SR5 and SR10) were treated with SrE (150 and 300 mg/kgb.w. daily) dissolved in olive oil. Tolerability and toxicity of SrE were estimated on the basis of daily monitoring of behavior, body weight gain (BWG), relative weight of liver, left kidney, prostate and left testis, and values of general biochemical parameters. Total liver proteins (TLP) and glutathione content in hepatocyte suspension were also determined. RESULTS: BWG was significantly unchanged in SR5 and SR10 compared to both controls in all intervals of measurement and at the end of treatment (p > 0.05). LW/BW ratio was significantly higher in SR10 compared with O (p < 0.01). Creatinine and potassium were significantly higher in SR5 compared to O (p < 0.05), but in SR10 were significantly higher compared to both control groups (p < 0.01). TLP content was significantly higher in SR5 compared to OO (p < 0.01). The content of glutathione in homogeneous suspension of hepatocytes didn't alter significantly. CONCLUSIONS: Obtained results have expanded the current state of knowledge about the tolerability and toxicity of SrE, in particular of Prostamol-unoR. For the adoption of a more precise conclusion about its tolerability and toxicity, it should be excluded possible limiting factors that we identified in this study.

Arbutin content and antioxidant activity of some Ericaceae species.

Quantitative analyses and investigation of antioxidant activity of herb and dry ethanolic extracts of five species from Ericaceae family (Arbutus unedo L., Bruckentalia spiculifolia Rchb., Calluna vulgaris Salisb., Erica arborea L. and Erica carnea L.) were performed. Total polyphenols, tannins and flavonoids were determined spectrophotometrically and arbutin content was measured both spectrophotometrically and by HPLC coupled with DAD detection. Antioxidative properites of the ethanolic extracts were tested by means of FRAP (total antioxidant capacity), lipid peroxidation and DPPH free radical scavenging activity. A significant amount of arbutin was detected only in Arbutus unedo. All samples investigated showed excellent antioxidant activity. The best inhibition of lipid peroxidation has been shown by Bruckentalia spiculifolia herb extract (62.5 microg/ml; more than 95%), which contained the highest amount of flavonoids (11.79%). The highest scavenging activity was obtained with leave extract of Arbutus unedo (IC50 = 7.14 microg/ml). The leaves of A. unedo contained a small amount of flavonoids but high content of non-tannins polyphenols.

Development of a high resolution three-dimensional surgical atlas of the murine head for strains 129S1/SvImJ and C57Bl/6J using magnetic resonance imaging and micro-computed tomography.

The mouse has emerged as a major experimental model system for examining the functional properties of the mammalian CNS; both during development and following CNS injury. Histologic procedures currently used to determine the relative position of structures within the CNS are presently limited in their ability to take full advantage of this system for surgical and morphometric procedures. We present here the first three-dimensional interactive digital atlas of the murine brain and skull for two genetically important strains of mice; 129S1/SvImJ and C57Bl/6J. The final resolution of these digital atlases is 54 micro m(3). These representations of the murine brain and skull, in conjunction with our development of a new, more dynamic master coordinate system, provide improved accuracy with respect to targeting CNS structures during surgery compared with previous systems. The interactive three-dimensional nature of these atlases also provide users with stereotactic information necessary to perform accurate "off-axis" surgical procedures, as is commonly required for experiments such as in vivo micro-electroporation. In addition, three-dimensional analysis of the brain and skull shape in C57Bl, 129Sv, CD1, and additional murine strains, suggests that a stereotactic coordinate system based upon the lambda and rostral confluence of the sinuses at the sagittal midline, provides improved accuracy compared with the traditional lambda-bregma landmark system. These findings demonstrate the utility of developing highly accurate and robust three-dimensional representations of the murine brain and skull, in which experimental outputs can be directly compared using a unified coordinate system. The aim of these studies is to enhance comparative morphometric analyses and stereotactic surgical procedures in mice.

Evaluation of the anti-inflammatory and cytotoxic activities of naphthazarine derivatives from Onosma leptantha.

The root extracts of Onosma leptanhtha were evaluated for their anti-iflammatory and cytotoxic activities. The cyclohexane extract, which appeared as the most active in both assays, has been further subjected to bioassay-directed fractionation to afford the naphthazarine derivatives: beta,beta-dimethylacrylshikonin (1), isovalerylshikonin (2) and acetylshikonin (3). The evaluation of the anti-inflammatory activity was performed on carrageenan-induced rat paw edema test. All the tested compounds proved to be active, while compound 3 showed the best anti-inflammatory effect. In addition, the cytotoxic activity of the extracts and isolated compounds, was also assayed against L1210 murine lymphoblastic leukemia cell line, and human fibrosarcoma HT-1080 cells. Compound 1 exhibited remarkable cytotoxic activity (390 nM for L1210 cells), which is superior to that of shikonin, which was used as control.

Cytotoxic and antioxidant activity of Achillea alexandri-regis.

The cytotoxicity and antioxidant properties of herb extracts of Achillea alexandri-regis were studied. Combined chloroform and ethylacetate extracts exhibited a pronounced cytotoxic effect against HeLa cancer cells (IC50 = 25.92 +/- 4.96 microg/ml), and lower cytotoxicity against K562 leukemia cells (IC50 = 48.59 +/- 18.31 microg/ml). The methanol extract was found to be a moderately cytotoxic in vitro agent against HeLa and K562 cells. No suppressive activity was detected on non-malignant peripheral blood mononuclear cells (PBMC). The antioxidant activity of the methanol extract was assessed by DPPH radical scavenging. The methanol extract of A. alexandri-regis showed concentration dependent DPPH radical scavenging activity with IC50 = 36.14 +/- 0.05 microg/ml.

Quantification of allantoin in various Zea mays L. hybrids by RP-HPLC with UV detection.

A RP-HPLC method for quantification of allantoin in silk of fifteen maize hybrids (Zea mays L., Poaceae) was described. Following extraction of the plant material with an acetone-water (7:3, VN) mixture, filtration and dilution, the extracts were analyzed without previous chemical derivatization. Separation and quantification were achieved using an Alltech Econosil C18 column under isocratic conditions at 40 degrees C. The mobile phase flow (20% methanol--80% water with 5 mM sodium laurylsulfate added at pH 2.5, adjusted with 85% orthophosphoric acid; pH of water phase was finally adjusted at 6.0 by addition of triethylamine) was maintained at 1.0 mL/min. Column effluent was monitored at 235 nm. This simple procedure afforded efficient separation and quantification of allantoin in plant material, without interference of polyphenols or other plant constituents of medium to high polarity, or similar UV absorption. Our study revealed that the silk of all investigated maize hybrids could be considered relatively rich in allantoin, covering the concentration range between 215 and 289 mg per 100 g of dry plant material.

Diuretic activity of Maydis stigma extract in rats.

Maydis stigma (corn silk) is a herbal drug reputed for the treatment of urinary ailments in various traditional medicine systems. To determine its influence on urinary volume and the excretion of sodium, potassium and chloride, 5% and 10% decoctions were administered daily to adult male Wistar rats for eight days. The concentration of electrolytes and urea in plasma, the influence of treatment on urinary pH value as well as creatinine clearance were also investigated. Daily oral administration of 5% decoction at the dose of 10 ml/kg led to a significant and acute diuresis in rats, reaching the peak value in the first 24 h of treatment. Over a similar period, application of 10% decoction did not affect urinary excretion of water, but significantly increased the pH value of excreted urine. A significant decrease in sodium and chloride plasma levels was observed in both treated groups. The creatinine clearance was markedly increased after the treatment with both extracts. Our findings indicate that the diuretic effect of 5% aqueous Maydis stigma extract is in accordance with the increase in glomerular filtration rate and inhibition of sodium and chloride tubular reabsorption, caused a by still unidentified intrinsic factor, but not the salt-loading effect.

[Gastrointestinal stromal tumor of small intestine--case report]. / Gastrointestinalni stromalni tumor tankog creva.

Gastrointestinal stromal tumor is a form of mesenchimal neoplasm that may be present in all parts of gastrointestinal system. We are reviewing diagnostic and therapeutic algorithms for patient with diagnosed gastrointestinal neoplasm of small intestine, experiencing repeated episodes of painful bleeding from gastrointestinal trackt.

[Sodium excretion in children with lithogenic disorders]. / Izlucivanje natrijuma kod dece s litogenim poremecajima.

INTRODUCTION: The causes of nephrolithisis are multifactorial and have not yet been enough investigated [1]. Hypercalciuria is the most common cause of metabolic nephrolithiasis [2-4]. Close relationship between urinary calcium and urinary sodium has been a subject of reported observations in the past, showing that high urinary sodium is associated with high urinary calcium [5-7]. Hyperoxaluria, hyperuricosuria and cystinuria are also metabolic disorders that can lead to nephrolithiasis. Recent studies have indicated that urinary elimination of cystine is influenced by urinary sodium excretion. Based on these observations it has been hypothesised that patients with high urinary sodium excretion are at high risk of urinary stone disease. The purpose of the study was to investigate sodium excretion in a 24-hour urine and first morning urine collected from children with lithogenic metabolic abnormalities (hypercalciuria, hyperoxaluria, hyperuricosuria, cystinuria), both with nephrolithiasis and without it, in order to determine its significance in urinary calculi formation. PATIENTS AND METHODS: Urinary sodium excretion was investigated in 2 groups of children: patients with lithogenic metabolic abnormalities, but without urinary stone disease (L group) and patients with nephrolithiasis (C group). Both groups were divided into 2 subgroups: patients with hypercalciuria and without it. There were 22 patients in group L (mean age 11.97 +/- 4.13 years), of whom 17 formed a hypercalciuric subgroup and 5 formed a non-hypercalciuric subgroup (3 patients with hyperuricosuria and 2 patients with hyperoxaluria). Group C consisted of 21 patients with nephrolithiasis (mean age 12.67 +/- 3.44 years), of whom 6 formed a hypercalciuric subgroup and 15 formed a non-hypercalciuric group (2 patients with cystinuria and 13 patients without lithogenic metabolic abnormalities). Control group consisted of 42 healthy age-matched children. All subjects had a normal renal function. A detailed history and clinical examination were done, and ultrasonography was performed in all patients. A 24-hour urine, first morning urine and serum specimen were analysed for sodium, potassium, calcium, uric acid, urea and creatinine. Fractional excretion of sodium, as well as urinary sodium to creatinin ratio and urinary sodium to potassium ratio, were calculated from the findings. Sodium and potassium levels were determined by flame photometry, calcium was measured by atomic absorption technique (Beckman Atomic Spectrophotometer, Synchron CX-5 model, USA), uric acid by carbonate method and creatinine by Jaffe technique. Cystine and dibasic amino acids were quantified by ion chromatography. Urinary oxalate excretion was determined by enzyme spectrophotometry. Hypercalciuria was defined by 24-hour calcium excretion greater than 3.5 mg/kg per day and/or calcium to creatinine ratio greater than 0.20 [8]. Uric acid excretion was expressed as uric acid excretion factored for glomerular filtration, according to Stapleton's and Nash's formula [9]. Normal values were lower than 0.57 mg/dl of glomerular filtration rate in 24-hour samples. Mean values were statistically analyzed by Pearson's linear correlation and analysis of variance (ANOVA). RESULTS: Urinary sodium concentration values including urinary sodium to potassium ratios, are shown in Table 1. We found that urinary sodium excretion was significantly increased in patients of both L and C groups when compared with controls (p < 0.05). Further analysis of the subgroups showed that urinary sodium excretion was significantly higher only in patients with hypercalciuria of both L and C groups in comparison to controls (p < 0.05) (Table 2). A significant positive correlation was found between 24-hour urinary sodium to creatinine ratio and urinary calcium to creatinine ratio (r = 0.31; p < 0.001) (Graph 1), as well as between urinary sodium to potassium ratio in 24-hour and first morning urine (r = 0.69; p < 0.001) (Graph 2). (A

A model of in vivo dosimetry and quality assurance analysis of total body irradiation in Zagreb.

The aim of the study was to develop a systemic and simple method for determination of midplane dose factors and lung correction factors, suitable for semiconductor 60-Co total body irradiation (TBI) in vivo dosimetry, based on input (entrance) and output (exit) dose readings. A theoretical model of anthropomorphic phantoms of cylindrical ellipsoid shape made it possible to generalize the applicability to any individual case of the real patient's data. Also, the quality assurance analysis of dosimetric results of 230 patients who had undergone TBI during a 10-year period, was performed. The ratio of the mean measured dose values for all locations in all patients and stated dose (12.45 Gy in 3 days, 3 fractions) was 0.998, SD 0.033. The upper limit of the total combined uncertainty of TBI midplane absorbed dose, including all locations and all sources of error, amounted to 9.4%.

[Surgical treatment of liver metastases of colorectal carcinoma]. / Hirursko lecenje metastaza kolorektalnog karcinoma u jetri.

Colorectal carcinoma metastasizes into the liver, but liver-only metastases are infrequent. Liver-only metastases are seen mainly from colorectal carcinoma. This is the only metastatic disease where treatment aimed only or mainly at the liver metastases is employed with curative intent. If liver resection for colorectal metastases is done by an experienced team, adhering to predefined indications, five year survival ranges from 30-40%, operative mortality is 3-5% and the postoperative morbidity is acceptable. New diagnostic techniques have been introduced and indications for liver resection extended. This paper presents the current limitations and possibilities for the surgical management of colorectal metastases in the liver.

[Spontaneous massive retroperitoneal hemorrhage from an adrenal gland cyst]. / Spontano masivno retroperitonealno krvavljenje iz ciste nadbubrezne zlezde.

Adrenal cyst are rare disease. Bleeding, particularly massive, from these cysts is even rarer. The cyst causing spontaneous massive retroperitoneal bleeding in a 17 year old girl is presented. Adrenalectomy was successfully carried out. The patient stayed symptom free so far.

[Echohistograms of circumscribed liver lesions]. / Ehohistogrami cirkumskriptnih lezija jetre.

The term focal liver changes usually applies to benign and malignant primary and secondary tumours. However, according to echotomographic findings, tumour-like diseases, such as focal nodular hyperplasia and diseases caused by bacteria, fungi, protozoa and parasites may also be included. Improvement in echotomographic technique and use of echohistogram have recently enabled provision of data determining the aetiology, i.e. pathohistological structure of changes with computerized analysis. The study included 178 patients with circumscribed liver lesions: thirty patients had primary carcinoma, 39 had secondary carcinoma, 41 had cystic disease, 35 hemangioma, 10 focal steatosis, 23 had other circumscribed lesions, as well as 175 subjects with healthy liver. The examinations were performed using the real-time ulstrasound apparatus (Toshiba SA 100 A). Echohistograms were obtained by placing certain amount of liver tissue onto X and Y axes, to be processed and graphically presented by the apparatus. X axis showed the number of particles (N) of liver tissue on certain surface, while Y axis showed the average value (M) and maximal value (Max) of particles. Analysis of echohistographic parameters evidenced the following: mean distribution values of M, Max and N differed in different pathological conditions (Tables 1 and 2). Correlation analysis of the studied parameters revealed different values among the studied groups of patients (Table 3, Figure 1). Analysis of echohistograms and their parameters revealed differences between the tissues of the "healthy" and "affected" liver which may be significant in diagnosis of circumscribed liver changes. N/Max:N/M ratio was higher in normal liver when compared to the studied groups of patients (except for N/M subration in focal changes). Our study, as well as the studies performed by aforementioned authors, evidenced close correlation of the appearance of echohistograms and parameter ratio with density, homogenicity and greatly with vascularization of the studied tissue. It has also been evidenced that the interrelations between the echosistographic parameters are in a complexly interwoven, and partly in aetiology, or it is better to say in relations between the healthy tissue and pathological liver changes. However, it must be concluded that the series of the described cases is small to enable establishment of diagnostic criteria exclusively based on echohistograms. Similar analysis of echohistograms in circumscribed liver diseases was not evidenced in the referential literature published so far. We consider the method useful and believe that the future development of computerized ultrasound extensions will enable differentiation of lesions.

[Gallbladder motor function studied by modified infusion cholescintigraphy method after gastric and duodenal surgery]. / Ispitivanje motorne funkcije zucne kese modifikovanom infuzionom holescintigrafijom posle operacija na zelucu i duodenumu.

Estimation of the gallbladder (GB) motility disorders after gastric surgery has not yet been assessed because of the shortage of the reliable diagnostic methods. The aim of the study is introduction, modification and establishment of the infusion cholescintigraphy into clinical practice and its performance in the groups of patients with gastric resection (RVBI i RVBII), total gastrectomy and patients after gastroplasty. Obtained data produced information about motility disorders caused by billateral truncal vagotomy, lack of the food transit through duodenum and if the motility disorders are the same in different time periods after operation. In groups of patients without truncal vagotomy (RVBI and RVBII), minor motility disorders are registered in comparison to the groups after truncal vagotomy. In the period of 6 months after surgery, higher motility disorders are registered in the group of patients with total gastrectomy, while after 9-12 months GB motility completely recovers. Groups with preserved transit of food through duodenum (RVBI and gastroplasty), has minor motility disorders in comparison to the group without transit of food through duodenum (RVBII and total gastrectomy). By introducing infusion cholescintigraphy, reliable method for the GB motility assessment is obtained.

[Retroperitoneal sarcoma]. / Retroperitonealni sarkomi.

Clinical experience in the treatment of retroperitoneal soft tissue sarcomas during period 1978-1987 is presented. The material consists of 64 patients with the male predominance (60.94%), and the peak occurrence in the forth and fifth decade of life. The difficulties in the diagnostic of these tumours are mostly caused by unspecific clinical picture, which is usually cleared with the stage of inoperability. Modern diagnostic approach, using echotomography and angiography enables earlier diagnosis. The most often seen histological pictures in this series are: M. Hodgkin (28.14%), Liposarcoma (25%) Lympho and fibrosarcoma (twice 12.5%), and the other forms are rarely encountered (1-3%). The operability is very low, and the radical operation is performed only in 16 cases (25.50%), and exploratory procedures are performed in 44 (68.75%). In the rest of cases a reduction of bulk mass of tumour was performed. The 5 year survival in the radically operated is present in 25% (4 patients) what is in accordance with the literature. In the cases with exploration or partial resection, the average survival is 4.5 months, in spite of some longer survivals in the group with Hodgkin's disease, mostly because of the modern cytostatic treatment. Intermediate postoperative mortality is 3.12% (2 patients). In recent years, the more aggressive surgical approach is advised, resulting in better outcome.