Native Non-Saccharomyces Yeasts as a Tool to Produce Distinctive and Diverse Tamjanika Grape Wines.

The enological potential of two previously characterized indigenous yeast isolates, Hanseniaspora uvarum S-2 and Candida famata WB-1, in pure and sequential inoculation with commercial yeast Saccharomyces cerevisiae QA23 were analyzed in industrial-scale vinification of the grape variety Tamjanika. Their contribution to the quality and aroma profile was investigated by quantifying volatile compounds and wine sensory evaluation. Both yeast isolates were able to complete alcoholic fermentation, to reduce ethanol concentration up to 1.06% v/v (in monoculture) in comparation to S. cerevisiae QA23, and to enhance aroma and sensory profile. Based on calculated odor activity values (OAV), p-cymene, ethyl hexanoate, ethyl octanoate, and ethyl decanoate were the major aroma volatile compounds in all Tamjanika wine samples. Analyzed yeast strains significantly affected relative contribution of volatile compounds and can be considered responsible for the differences and uniqueness of the obtained wine samples. Besides confirmation of good enological and fermentative characteristics, selected isolates can be characterized as high ester-producing strains with potential to enhance the floral and fruity aromas of wine. The present study represents a further step toward the use of indigenous yeast isolates at industrial-scale fermentation in order to ensure the regional signature of Tamjanika wine.

Characterization of free and insoluble-bound phenolics of chia (Salvia hispanica L.) seeds.

The content, antioxidant activity and composition of free and bound phenolics from chia seeds were investigated. The free phenolics were extracted by using ethanol and methanol of analytical purity, 70% v/v ethanol and by 70% v/v methanol. The bound phenolics were extracted from seeds after alkaline hydrolysis was done. The phenolics content obtained by aqueous-alcoholic solutions was higher than the content obtained by using the pure alcoholic solution (for 20.8% by 70% methanol and 41.2% by 70% ethanol). The content of bound phenolics was 932 µg g-1. Among the identified phenolic compounds the apigenin 4'-O-glucoside was the most abundant, and among the acids, a rosmarinic was the most abundant in free phenolic extract and caffeic acid in bound. The correlation coefficients indicated the DPPH method and reducing power were more suitable for assessing the antioxidant activity than the FRAP method.

An insight into chemical composition and bioactivity of 'Prokupac' red wine.

Autochthonous variety 'Prokupac' (Vitis vinifera L.) red wines with the addition of selected aromatic herbs (Artemisia absinthium, Cinnamomum zeylanicum, Glycyrrhiza glabra and Pimpinella anisum L.) were evaluated for chemical composition and biological activity. Furthermore, their polyphenolic profiles were determined by HPLC-DAD. While total phenolic (TPC) and flavonoid (TFC) contents, along with antioxidant activity, were significantly higher in all examined wine samples, the content of total anthocyanins was decreased, compared to a pure 'Prokupac' wine, used as the control one. On the other hand, 'Prokupac' wines with the addition of cinnamon (S. zeylanicum) and wormwood (A. absinthium) displayed highest antioxidant (EC50, 0.021 ± 0.001 mg/mL) and antibacterial (Enterococcus faecalis, MIC/MBC 15.63/62.50 µL/mL) activities, respectively. The study showed that 'Prokupac' wine with the addition of cinnamon was significantly enriched both with TPC and TFC, compared to control wine (p < 0.05), which may contribute to its market potential.

Immunohistochemical expression of proliferative markers in renal cell carcinoma.

PURPOSE: The purpose of this study was to investigate into the expression of cyclin A and telomerase in renal cell carcinoma (RCC) and to analyze the relationship between expression and the clinicopathological characteristics of the tumor and their impact on survival. METHODS: The overall material included 74 samples of RCC and 4 of normal renal tissue. Primary cyclin A antibody from Santa Cruz Biotechnology and TERT MA5-16034 antibody from Thermo Fisher Scientific Inc were used. Staining was performed by streptavidin-biotin technique using DAKO LSAB+ kit. Statistical analyses were performed using of SPSS 23 Statistics software from IBM. RESULTS: No differences in cyclin A and telomerase expression among gender and age groups were found, nor did the tumor dimensions have any significant impact on expression. Also, tumor grades and stages did not differ. However, histological types differed in favor of the papillary type. A significant positive correlation between both markers, as well as between the expression and tumor stage and grade was noticed. Only the tumor stage had negative impact on survival. CONCLUSIONS: Although not affecting survival, the expression of cyclin A and telomerase increased with tumor stage and grade, suggesting that cyclin A and telomerase could be potential proliferative immunohistochemical markers of RCC.

Immunohistochemical study of cyclin A and p16 expression in patients with renal cell carcinoma.

PURPOSE: Renal cell carcinoma (RCC) is the most common malignant kidney tumor in adults. Dysregulation of the cell cycle can lead to cancer development. In this study, the mitosis-associated cyclin A and p16, a negative controller, were investigated as potential key points in the RCC development. METHODS: This retrospective study included 74 patients with RCC. The expression of cyclin A and p16 and their correlation to histopathological parameters (TNM stage, histological subtype, nuclear grade, tumor size), gender, age, and clinical outcome were studied and analyzed. RESULTS: The highest median value for cyclin A (40%; range 0-70)) and for p16 (57.5%); range 35-80) were found in the papillary histological subtype. Survival analysis showed that in the group of patients that had died before September 2015, the median value for cyclin A was 20% (range 0-60), which was significantly higher than 5% (range 0-70), found in the group of patients that survived (p=0.019). CONCLUSIONS: In relation to the histological subtype, the papillary type of RCC was associated with a significantly higher expression of cyclin A and p16 compared to other subtypes of RCC. High expression of cyclin A indicated worse prognosis, therefore cyclin A could be considered to be a significant prognostic marker.

Polymorphisms of survivin -31 G/C gene are associated with risk of urothelial carcinoma in Serbian population.

PURPOSE: Survivin is thought to play an important role in carcinogenesis and is found to be associated with poor clinical outcome in various malignancies. Gene -31 G/C polymorphism has been identified as a risk factor for the development of several types of tumors. The purpose of this study was to investigate the association between survivin gene promoter -31C/G polymorphism and urothelial carcinoma (UC) risk in Serbian population and to compare the different expressions of survivin in UC of different disease stages, histological grades and tumor location in the upper or lower urinary tract. METHODS: DNA from 94 patients with primary UC and from 82 healthy subjects was subjected to PCR restriction fragment length polymorphism analysis (PCR-RFLP) to identify individual genotypes. UC samples were subjected to immunohistochemical analysis to assess survivin expression in these lesions. RESULTS: It was observed that the frequency of G/G genotype was greater in patients with UC (58.7%) than in controls (32%). Compared with study subjects carrying the C/G or C/C genotypes, significantly increased UC risk was found for individuals carrying the G/G genotype. Those carrying the G/G genotype had a significantly increased UC risk compared with those with C/G or C/C genotypes. Patients with UC carrying the G/G genotype had a greater prevalence of muscle-invading (stage T2-T4), high-grade (G2) tumor and immunohistochemicaly overexpressed survivin compared with those carrying the C/G or C/C genotypes. CONCLUSIONS: G/G genotype of the -31C/G polymorphism might be a risk factor for UC development.

Stenting versus non-stenting following uncomplicated ureteroscopic lithotripsy: Comparsion and evaluation of symptoms.

Background/Aim: Currently, ureterorenoscopic (URS) stone fragmentation and removal is the treatment of choice for managing ureteral stones, especially mid and distal ones and is advocated as initial management of ureteric stones. The aim of this work was to evaluate the symptoms, necessity, potential benefits and adverse effects of ureteral stent placement after uncomplicated ureteroscopic lithotripsy. Methods: This retrospective-prospective study evaluated a total of 125 patients who had underwent ureteroscopic lithotripsy (URSL). The patients were divided into two groups: stented (59 patients) and unstented (controls, 66 patients). The outcomes measured and compared between the two groups included: stone free rate, postoperative patient pain validated by scale, lower urinary tract symptoms (LUTS), the need for unplanned hospital care, stent related complications, and functional recovery in the form return to normal physical activities. Results: A successful outcome, defined as being stonefree after 12 weeks, was achieved in all 125 (100%) patients. The stone-free rate showed no significant differences between the two groups. LUTS was frequent complaint in the stented group, with statistically significant difference in the domain of frequency/urgency (p = 0.0314). There was a statistically significant difference between the groups in the mean operative time and mean hospitalization time, mean pain visual analog scale (VAS) score and in the use of nonnarcotic analgesic. On the day of the surgery and until postoperative day 3 (POD 3) and postoperative day 5 (POD 5), the pain score was much higher among stented patients than among the controls (p = 0.0001) and non-narcotic analgesic use (p = 0.001) was frequently required in the stented group. Conclusion: Routine placement of ureteral stent after URSL is not mandatory and may be associated with stent side effects. Uncomplicated URSL is safe without stent placement after the treatment.

The clinical course of non-muscle invasive bladder cancer after transurethral resection of the tumor with or without subsequent intravesical application of bacillus Calmette-Guérin: The influence of patients gender and age.

UNLABELLED: BACGROUND/AIM: The therapy with intravesical instillation of bacillus Calmette-Guérin (BCG) after transurethral resection (TUR) of tumor is the gold standard of treatment of non-muscle invasive bladder cancer (NMIBC). The role and importance of BCG intravesical therapy in various shape of tumors, were confirmed by our previous investigation. The aim of this study was to examine whether incidence of recurrence and tumor regression differs depending on sex and age of patients. METHODS: This study included a total of 899 patients suffering from NIMBC, treated at our institution from January 1, 2007 to March 1, 2013. Two groups of patients were formed: patients underwent TUR + BCG therapy (the group I) and the group II with patients in whom TUR was performed as only therapy. These two groups of patients were divided into subgroups of respondents male and female, age 60 years or younger and older than 60 years. Statistical analysis was performed using χ2 test and the Kolmogorov-Smirnov test. RESULTS: This research suggests that if the frequency of recurrence is seen as the only parameter, considering all the subjects, the lowest recurrence rate was determined in the male subjects, aged 60 years and younger who had received BCG after TUR. A high statistical significance was found in the incidence of recurrence in patients younger than 60 years, depending on the response to the therapy, while in those older than 60 years, the difference was at the level of statistical significance. This can be attributed to a certain degree of infravesical obstruction in older men. CONCLUSIONS: Sex and age of patients may have a significant influence on the course and outcome of NMIBC. The disease has the most malignant and most aggressive behavior when present in males older than 60 years.

Difference in recurrence frequencies of non-muscle-invasive-bladder tumors depending on optimal usage of intravesical immunotherapy of bacillus Calmette-Guérin.

BACKGROUND/AIM: The therapy with intravesical instillation of bacillus Calmette-Guérin (BCG) after transurethral resection (ITJR) of the tumor is the gold standard of treatment of non-muscle invasive bladder cancer (NMIBC). The aim of this study was to compare the frequencies of reccurence between a group of patients submitted to TUR + BCG therapy (group I) and a group of patients submitted only to TUR (group II). METHODS: The patients with NMIBC, a total of 899, treated in our Institution from January 1, 2007 to March, 2013, were included in this study and divided into two groups: group I and group II. These two groups were divided into three subgroups: solitary first diagnosed tumor ≤ 3 cm (SFDGT), solitary first diagnosed tumor > 3 cm and multiple first diagnosed tumors (MFDGT), and recedive tumors (RCT). Statistical analysis was performed by using χ2-test and Kolmogorov-Smirnov test. RESULTS: In the group I a total of 133 cases had reccurence contrary to 75 in the group II, making a statistically highly significant difference. Analysis of recurrences through the subgroups revealed: in the group I SFDGT recurrence occured in 27 of the cases vs 9 cases in the group II; in the group I MFDGT recurrence occured in 49 of the cases vs 31 in the group II (p < 0.001), and finally, in the group I RCT recurrence occured in 57 cases vs 35 cases in the group II (p < 0.001). CONCLUSION: The obtained results indicate no difference in the frequency of reccurence between the group I and group II regarding SFDGT, but a very high significant difference regarding those with MFDGT and RCT. These results should be taken into consideration in everyday clinical practise.

A characterization of content, composition and scavenging capacity of phenolic compounds in parsnip roots of various weight.

The contents, composition and radical scavenging capacity of phenolic compounds from parsnips of various root weights grown in Serbia were examined. The content of phenolic compounds depended on root size, and the highest content was in the group where the samples with the lowest root size were grouped. The content of phenolic compounds varied from 109.7 to 125.3 micromol of chlorogenic acid per g dry extract, i.e.5470.8 to 6245.1 mg per kg fresh weight. Among the identified compounds were quercetin, kaempferol, apigenin and genkwanin glucosides and hydroxycinnamic and caffeoylshikimic acids. The highest antioxidant capacity was found for the group with the lowest root weight; the IC50 values ranged from 1.59 to 2.49 mg mL(-1). There was positive correlation between the total phenolic content and scavenging capacity, and the kaempferol glucosides content could be an indicator of DPPH scavenging capacity of parsnip roots.

Management of calculus anuria using ureteroscopic lithotripsy as a first line treatment: its efficacy and safety.

PURPOSE: To present our experience with emergency ureteroscopic lithotripsy (URSL) for ureteral calculi associated with acute kidney injury (AKI). MATERIALS AND METHODS: We retrospectively evaluated the 61 patients consisted of 90 ureteral units (UU), who underwent URSL. The cause of anuria was bilateral calculus obstructions in 29 cases, and unilateral calculus obstruction with, absent, nephrectomized contralateral kidney in 32 cases. In the case of bilateral synchronous ureteric calculi same-session bilateral ureteroscopy (SBBU) was done. The duration of anuria varied between 12 to 72 hours. At the end of the procedure, ureteral stent was systematically left in place in all patients. Surgery was performed 6-12 hours after admission to hospital. Patients were followed at least 1 month postoperatively. RESULTS: The stone free rates (SFR) were determined as baseline, on the first post-operative day, and as overall on the 30 days after procedure. The greatest success was achieved in the distal localization of stones up to 10 mm (93%). Renal function returned in 51 (83.6%) patients within 7 days. In 18 (29.5%) patients [18 (20%) UU] we performed second procedure as extracorporeal shockwave lithotripsy in 16.7% and open surgery in 2.2%. In 43 (70.5%) patients URSL was a successful therapeutic approach in dealing with pain, obstruction and calculus. CONCLUSION: Calculus anuria is a medical emergency that requires rapid diagnosis and prompt treatment for the purpose of decompression. URSL is the proper method of choice for selected patients and can be performed safely and has high success rates with minimal morbidity.

Rare locations of metastastatic renal cell carcinoma: a presentation of three cases.

INTRODUCTION: Metastatic renal cell carcinoma (RCC) frequently spreads not only to neighboring lymph nodes, but also to distant organs, including the lungs, liver, bones and brain. CASE REPORT: We presented three cases of RCC with colon metastasis. In the first, 63-year-old patient, after left nephrectomy followed with lyphadenectomy in paraaortic lymph node, left hemicolectomy was done due to RCC metastasis in rectosigmoid colon. In the second, 35-year-old patient, left radical nephrectomy was followed two years later with partial right nephrectomy, lung metastasectomy, small bowel and coecum resection and right orchiectomy all as separate procedures in different time intervals. The patient died from brain and bone metastases two years after the first surgery. The third, 35-year-old patient, had right nephrectomy followed by repeted lymphadenectomies after 6, 12 and 24 months. Four years later RCC spreaded to coecum and right hemicolectomy was performed. CONCLUSION: RCC treated with nephrectomy should be carefully followed up with imaging methods as a proper treatment of RCC metastases to distant organs could be important for a patient survival.

Congenital arteriovenous fistula of the horseshoe kidney with multiple hemangiomas.

INTRODUCTION: Congenital renal arteriovenous fistulas (AVF) are rare, especially if they are associated with other developmental renal anomalies. CASE OUTLINE: A 34-year-old female was hospitalized due to total painless hematuria and bladder tamponade. Excretory urography revealed a horseshoe kidney with normal morphology of pyelocaliceal system and ureters. Aortography and selective renovasography detected a cluster-like vascular formation with multiple arteriovenous fistulas (AVF). Due to a large AVF gauge and poor flow of the efferent vein to the inferior vena cava, a surgical procedure of two renal artery segmentary branches ligation and division was performed. During the operative procedure, the presence of multiple superficial renal hemangiomas was detected. CONCLUSION: Although selective arterial embolization represents the preferable treatment option, conventional surgery remains favorable alternative in selected cases with large and complex AVF.

Expression of neural cell adhesion molecule in renal cell carcinoma.

Neural cell adhesion molecule (NCAM) is important for cell migration and it could be expressed in some renal cell carcinoma (RCC). In recent decades, the incidence of RCC has been steadily rising by 2-4% each year. In this study NCAM expression and correlation with nuclear grade in different RCC were analyzed. We analyzed NCAM expression on 7 different RCC cell lines and 32 different RCC by immunohistochemistry, immunofluorescence, Western blot and FACS analysis. NCAM expression is detected in 6 cell lines and 16 RCC cases. NCAM-140 kDa isoform is expressed in different RCC and RCC cell lines. NCAM expression in non-invasive clear cell RCC is lower than in clear cell RCC with high nuclear grade. Expression of NCAM is not exclusive for specific RCC type, so NCAM can not be used as a specific diagnostic marker for RCC. NCAM expression is in correlation with nuclear grade in clear cell RCC, suggesting that NCAM expression is involved in aggressive behavior and metastatic potential in RCC.

Ultrasound-assisted extraction of total phenols and flavonoids from dry tobacco (Nicotiana tabacum) leaves.

Yields of extracted substances, as well as total phenol and flavonoid compounds obtained by classical and ultrasonic extractions from dry leaves of two tobacco types (oriental and Virginia) by two different solvents (acetone and methanol) at two operating temperatures (25 and 40 degrees C) were compared. The yield of extractive, as well as total phenol and flavonoid compounds depended on the type of solvent, operational temperature and the tobacco type. The importance of these factors was assessed using 2(4) full factorial experiments without replication.

The preoperative activity of Th1 and Th17 cytokine axes in prediction of sepsis after radical cystectomy.

The aim of the study was to correlate the preoperative activity of Th1 and Th17 cytokine axes with the development of sepsis after radical cystectomy. The study involved twenty patients with the infiltrative transitional cell carcinoma of the urinary bladder without previous radiotherapy/chemotherapy, who underwent open radical cystectomy with urinary diversion. Preoperative plasma concentrations of Th1 cytokines interleukin 12 (IL-12) and interferon gamma (IFN-γ), and Th17 cytokines IL-23 and IL-17, were measured using ELISA. Preoperative expression of mRNA for IL-12p35, IFN-γ, IL-23p19 and IL-17 was quantified by real-time RT-PCR using mRNA extracted from peripheral blood mononuclear cells. Eight patients developed postoperative sepsis, diagnosed within two weeks post-operation as systemic inflammatory response syndrome in the presence of local or systemic infection. The preoperative basal plasma concentrations of Th1 and Th17 cytokines were slightly above the detection limits, with a tendency toward lower concentrations in patients who developed sepsis, but the difference was not significant (p>0.05). The preoperative expression of mRNA encoding IL-12p35 and IL-17 was significantly lower in patients who developed sepsis (p=0.003 and p=0.028, respectively). The similar trend was observed for IL-23p19 and IFN-γ, but the differences did not reach the statistical significance (p=0.051 and p=0.172, respectively). These data suggest that determination of preoperative Th1 and Th17 cytokine mRNA levels might be useful in predicting sepsis development after radical cystectomy.

Modeling the kinetics of calcium hydroxide catalyzed methanolysis of sunflower oil.

The kinetics of Ca(OH)(2)-catalyzed methanolysis of sunflower oil was studied at a moderate temperature (60 degrees C), a methanol-to-oil molar ratio (6:1) and different catalyst amounts (from 1% to 10% based on oil weight). The methanolysis process was shown to involve the initial triglyceride (TG) mass transfer controlled region, followed by the chemical reaction controlled region in the latter period. The TG mass transfer limitation was caused by the low available active specific catalyst surface due to the high adsorbed methanol concentration. Both the TG mass transfer and chemical reaction rates increased with increasing the catalyst amount.

Synchronous and multiple transitional cell carcinoma of the bladder and urachal cyst.

Incomplete involution of the allantoic duct can result in different pathological forms of urachus which can give rise to inflammation or late malignant changes. Among urachal tumors, adenocarcinoma is most frequent, although other histological types can also be found. The synchronous presentation of a urachal transitional cell tumor, along with recurrent superficial bladder tumors has not been reported previously. We are reporting a 49-year-old male patient in whom transitional cell carcinoma of a urachal cyst was found with recurrent, multiple bladder tumors. The diagnosis of urachal cyst tumor was established according to ultrasonography and computed tomography. Most of the bladder tumors were resected transurethrally while open surgical excision of the urachal cyst with en bloc resection of the bladder dome was performed. Recurrent bladder tumors were afterwards treated with Bacillus Calmette Guerin (BCG) instillations. A year after surgery the patient has no signs of local recurrence or distant metastases of transitional cell carcinoma.

Impact of carboxymethylcellulose on morphology and antibiotic production by Streptomyces hygroscopicus.

A chemically defined media consisting of carboxymethylcellulose (CMC) was developed to maximize the production of antibiotics, hexaene H-85 and azalomycine, by Streptomyces hygroscopicus CH-7. The production of antibiotics by filamentous organisms is often dependent on the morphology and size distribution of the pellet population within the culture. By adding the polymer to the fermentation medium, the growth was changed from a single large glob to small reproducible pellets, and wall growth was diminished to a minimum. Maximum concentrations of hexaene H-85 (146.7 mg/dm(3)) and azalomycine (188.6 mg/dm(3)) were reached at 3.0% and 1.0% (w/v) CMC, respectively.

Kinetics of sunflower oil methanolysis at low temperatures.

The kinetics of the sunflower oil methanolysis process was studied at lower temperatures (10-30 degrees C). The sigmoidal kinetics of the process was explained by the mass transfer controlled region in the initial heterogenous regime, followed by the chemical reaction controlled region in the pseudo-homogenous regime. A simple kinetic model, which did not require complex computation of the kinetic constants, was used for simulation of the TG conversion and the FAME formation in the latter regime: the fast irreversible second-order reaction was followed by the slow reversible second-order reaction close to the completion of the methanolysis reaction. The mass transfer was related to the drop size of the dispersed (methanol) phase, which reduced rapidly with the progress of the methanolysis reaction. This was attributed to the formation of the emulsifying agents stabilizing the emulsion of methanol drops into the oil.