Identifying prognostic parameters related to surgical technique in patients treated by robotic radical prostatectomy.

INTRODUCTION AND OBJECTIVE: The most frequently studied factors in patients treated by robotic radical prostatectomy are PSA and pathological features of the biopsy and prostatectomy specimen. Studies on the factors associated with the surgical technique are scarce and with controversial results. The objective is to identify all possible surgical factors and their relationship with disease-free and metastasis-free survival. PATIENTS AND METHOD: Prospective study approved by the Ethics Committee, including patients who underwent robotic radical prostatectomy since January 2009 with a minimum follow-up of 5 years. Surgeon, surgical time, blood loss, fascial access, continence techniques, preservation of the fascia, neurovascular bundles, bladder neck, urethra, learning curve and surgical complications, were analyzed as possible prognostic factors. We performed univariate and matched comparisons of survival using Kaplan-Meier estimation and long-rank tests. The significance level for multiple comparisons was established with False Discovery Rate-adjustment (adjusted p). RESULTS: Cohort of 667 patients with a median follow-up of 69 months. In univariate analysis, surgeon (adjp=0.018), preservation of puboprostatic ligaments (adjp=0.02), preservation of endopelvic fascia (adjp=0.001) and performing periurethral suspension (adjp<0.001) are poor prognostic factors for disease-free survival. Fascia preservation also negatively affects metastasis-free survival (adjp=0.04). Previous abdominal surgeries, prostate, surgical time, blood loss, type of residual urethra, middle lobe, fascial access, fascia or bladder neck preservation, have no statistical significance. CONCLUSIONS: The surgeon and specific aspects of the surgical technique are determining factors in disease-free survival. Preservation of the fascia is the only factor that negatively affects metastasis-free survival.

Effect of glycation on sodium caseinate-stabilized emulsions obtained by ultrasound.

This work explores the potential of high-intensity ultrasound to produce fine-dispersion, long-time-stable, oil-in-water emulsions prepared with native and glycated bovine sodium caseinate (SC). Regardless the ultrasound amplitude and time assayed, the sonicated emulsions of native SC at 0.5 mg/mL had much higher emulsifying activity indexes compared with those emulsions formed by Ultra-Turrax (IKA Werke GmbH & Co., Staufen, Germany) homogenization. Nevertheless, the native SC emulsions were very unstable despite the optimization of parameters such as protein concentration, amplitude of ultrasound wave, and sonication time by using a Box-Behnken design. Early glycation of SC with either galactose, lactose, or 10 kDa dextran substantially improved both emulsifying activity and the stability, whereas at advanced stages of glycation, SC emulsions showed notably reduced emulsifying properties, likely because extensive glycation of SC promoted its polymerization mainly through covalent cross-linking, as was demonstrated by particle size measurements. The increase in particle diameter of glycoconjugates likely affected the diffusion of SC from bulk to the oil-water interface and slowed the reorientation process of the protein at the interface. These findings show that the combined effect of early-stage glycation of SC and high-intensity ultrasound as an emergent technique to form emulsions has the potential to provide improved emulsions that could be used in several food applications.

Purification of lactulose from mixtures with lactose using pressurized liquid extraction with ethanol-water at different temperatures.

The viability of the purification of lactulose from a mixture with lactose [70:30 (w/w)] using pressurized liquid extraction (PLE) at 1500 psi for 30 min was studied. Different temperatures (from 40 to 130 degrees C) and proportions of ethanol:water (70:30, 80:20, 90:10, 95:5, and 100:0) as the extraction solvent were assayed. Lactose and lactulose were measured by gas chromatographic analysis as their trimethylsilyl derivatives. Data were fitted through multiple linear regressions to different quadratic models to describe both the extraction yield (in terms of mg of lactulose) and the purity of the lactulose extracted. The optimum extraction conditions provided by the model were as follows: extraction temperature, 40 degrees C; and solvent composition, 70:30 ethanol:water. PLE extraction under the optimized conditions was also applied to purify lactulose from lactose in a synthesis mixture. To our knowledge, this is the first time that PLE has been tested for extraction and purification of lactulose from its mixture with lactose; this technique showed several advantages over classical methods such as the short extraction time and the low solvent consumption.

[Importance of the forensic autopsy in the diagnosis of septic shock: a case report]. / Importancia de la autopsia forense en el diagnóstico del shock séptico: a propósito de un caso.

Septic shock sometimes starts with unspecific symptoms that hamper the clinical diagnosis and, therefore an appropriate treatment. When the septic shock follows a fulminating course with a fatal outcome, the etiological diagnosis has to be conducted post-mortem. Sudden unexpected deaths in children and young adults are frequently the object of medico-legal autopsies. Some sudden unexpected deaths have an infectious origin, which requires further analyses, including microbiology, to establish the cause of death. Here, the case of a fatal septic shock in a 19-month old male infant is presented. After a mild foot injury, an infection by Streptococcus pyogenes progressed to septic shock with a fatal outcome as post-mortem studies demonstrated.

Gemcitabine in the treatment of advanced transitional cell carcinoma of the urothelium.

M-VAC (cisplatin, methotrexate, adriamycin, vinblastine) combination chemotherapy has been for long time the standard of care in fit patient with advanced urothelial tumors. Gemcitabine/cisplatin with similar results and an improved toxicity profile has proved to be a new standard alternative. Whether or not we can improve survival with newer triplet regimens will depend upon the results of ongoing phase III trials. In addition to the new active drug combinations and targeted therapies, new approaches are emerging for treatment. Chemotherapy optimization using molecular markers predicting chemosensitivity are being applied. There is an obvious need to incorporate in clinical trials a systematic translational approach to explain both our successes and our failures.

Use of chitosan for selective removal of beta-lactoglobulin from whey.

A method is described for selective removal of undenatured beta-lactoglobulin from cheese whey based on interactions between whey proteins and chitosan. Whey was previously clarified at pH 4.5 with addition of chitosan (25 mg/100 mL), and selective removal of beta-lactoglobulin was studied in the pH interval 4.6 to 6.5. Addition of chitosan caused selective precipitation of beta-lactoglobulin that increased with pH. The content of beta-lactoglobulin in whey decreased as the amount of chitosan added was increased. At pH 6.2, addition of 1.9 to 3.0 mg/mL of chitosan led to complete removal of beta-lactoglobulin, whereas at least 80% of the rest of whey proteins remained in solution. The production of cheese whey without beta-lactoglobulin could help to expand the applications of dairy by-products in food processing, and to isolate hypoallergenic whey protein concentrates.

Selective recovery of glycosylated caseinmacropeptide with chitosan.

The use of chitosan, a partially deacetylated chitin, to fractionate aqueous solutions of caseinmacropeptides (CMPs) was studied. The polycationic character of chitosan at acidic pH values allows the formation of complexes with negatively charged CMP molecules, inducing their flocculation. Glycosylated CMP (GMP) has higher affinity for chitosan than nonglycosylated forms (NGMP). The carboxylic groups in the carbohydrate moiety of the GMP increase the negative charge of the molecule and may play a role in the selective precipitation. At pH 5.0, 0.08 mg/mL of chitosan completely removed the GMP whereas 70% of NGMP remained in solution. As the pH increased, the amount of chitosan to ensure complete removal of GMP increased up to 0.19 and 0.34 mg/mL for pH 6.0 and 6.6, respectively.

Improvement in right ventricular systolic function after surgical correction of isolated tricuspid regurgitation.

Chronic tricuspid regurgitation (TR) may lead to impairment in right ventricular (RV) function. Whether surgical correction results in restoration of normal RV geometry and function is unknown. The purpose of this study was to determine whether surgical correction of TR results in improved RV geometry and function. Measurements of RV areas were made from digitized 4-chamber echocardiographic views. Right ventricular end-diastolic volume (RVEDV), right ventricular end-systolic volume (RVESV), and ejection fraction (EF) were calculated with the single-plane subtraction method. There was a significant decrease in RVEDV (109.06 +/- 12.45 versus 71.63 +/- 6. 83; P =.005) and RVESV (76.2 +/- 9.83 versus 44.5 +/- 5.58; P =.002) and a significant increase in RVEF (0.30 +/- 0.05 versus 0.38 +/- 0. 05; P =.01) at a mean follow-up of 130 +/- 63 days after surgery. These results demonstrate significant remodeling of the right ventricle with reduction in size and improved EF after tricuspid valve surgery.

Early stages of maillard reaction in dehydrated orange juice.

The formation of furoylmethyl derivatives of amino acids as indicators of the early stages of Maillard reaction in dehydrated orange juices and model systems was studied. In stored dehydrated orange juices, the presence of furoylmethyl derivatives of arginine, asparagine, proline, alanine, glutamic acid, and GABA was detected. Their formation increased with temperature of storage. After 2 weeks at 30 degrees C and a(w) = 0.44, the reconstituted orange juice contained 94 mg/L furoylmethyl derivatives, whereas up to 1215 mg/L was detected in samples stored at 50 degrees C.

Presence of 2-furoylmethyl derivatives in hydrolysates of processed tomato products.

Acid hydrolysis of Amadori compounds yields the corresponding 2-furoylmethylamino acids (2-FM-AA) that can be analyzed by ion-pair HPLC. The relative proportions of the different 2-FM-AA present in the hydrolysates of tomato products were determined to assess their usefulness as indicators of quality. In the lyophilized tomato samples stored at 50 degrees C and a(w) = 0.44 the formation of 2-FM derivatives of alanine, gamma-aminobutyric acid (GABA), asparagine, aspartic acid, glutamic acid, lysine, serine, and threonine was detected. In commercial tomato products the most abundant 2-FM-AA was 2-FM-GABA (from traces to 26.4 mg/100 g of dry matter) followed by 2-FM-lysine (furosine). Differences in 2-FM-AA contents among samples may be related to processing and storage conditions.

Use of 2-furoylmethyl derivatives of GABA and arginine as indicators of the initial steps of maillard reaction in orange juice.

The formation of 2-furoylmethyl derivatives of GABA (2-FM-GABA) and arginine (2-FM-Arg) as early indicators of nonenzymatic browning in different types of orange juice was studied. In dehydrated orange juice, the presence of 2-FM-GABA and 2-FM-Arg was detected from the first day of storage at 30 degrees C. In this type of juice, the content of these two compounds increased with temperature (30, 50 degrees C) and time (1-7 days) of storage. A noticeable increase in 5-hydroxymethylfurfural was only observed after 4 days of storage at 50 degrees C. No formation of 2-FM-GABA and 2-FM-Arg was detected in liquid orange juice heated under conditions similar to those used in the industry. These furoylmethyl derivatives were also found in commercial orange juice made from concentrates. A slight increase in their concentration was observed in the two samples stored during 8 months at room temperature. According to the results obtained, 2-FM-GABA and 2-FM-Arg contents could be suitable indicators to assess the main modifications due to Maillard reaction produced during the manufacture and/or storage of orange juice concentrates.

Formation of Amadori compounds in dehydrated fruits.

The presence of Amadori compounds in commercial dehydrated fruits has been shown through HPLC analysis of the corresponding 2-furoylmethyl-amino acids obtained by acid hydrolysis. Furosine (2-furoylmethyl-lysine) was the main 2-furoylmethyl derivative observed in dried figs and apricot samples, whereas in prunes and dates similar amounts of furosine and 2-furoylmethyl-gamma-aminobutyric acid were detected. A considerable variation of 2-furoylmethyl-amino acid contents among commercial raisin samples was observed. 2-Furoylmethyl-gamma-aminobutyric acid and 2-furoylmethyl-arginine, the most abundant 2-furoylmethyl-amino acids, ranged between 9.9 and 75.8 mg/100 g sample and 10.0 and 62.5 mg/100 g sample, respectively. Most of the Amadori compounds present in raisins seem to have originated during the commercial shelf life period rather than during processing. Determination of 2-furoylmethyl-amino acids could be used as a method of controlling commercial dehydrated fruit and selecting storage conditions.

Synthesis and characterization of gamma-N-(2-furoylmethyl)aminobutyric acid.

The product of acid hydrolysis of the Amadori compound gamma-N-(1-deoxy-D-fructosyl)aminobutyric acid was isolated and identified by (1)H NMR and (13)C NMR as gamma-N-(2-furoylmethyl)aminobutyric acid. This compound is an analogue to furosine, formed during acid hydrolysis of the corresponding Amadori compound. The retention time of the isolated compound was the same as that of the main peak observed in acid hydrolysates of stored orange juice powder. gamma-N-(2-Furoylmethyl)aminobutyric acid can be a useful indicator of the early stages of Maillard reaction in foods containing free gamma-aminobutyric acid.

Survey of the furosine content in cheeses marketed in Spain.

The furosine content in 53 samples of different cheese types (fresh, ripened with molds, artisanal hard-pressed, industrial hard-pressed, and processed) marketed in Spain was analyzed. The lowest furosine values were observed in artisanal hard-pressed cheeses (4.8 to 10.2 mg per 100 g of protein) and ripened with molds cheeses (4.2 to 12.8 mg per 100 g of protein). Industrial hard-pressed cheeses showed furosine values between 3.5 and 43.8 mg per 100 g of protein. Differences between samples may be attributed to the heat-treatment intensity given to cheese milk and to cheesemaking conditions. The highest contents of furosine were observed in processed cheeses (20 to 366.6 mg per 100 g of protein). A wide range of furosine content was found in fresh cheeses (17.9 to 73.6 mg per 100 g of protein), which could be due to the different amounts of dried milk added during the manufacture of cheeses.

Salt balance and rennet clotting properties of cow's, ewe's, and goat's milks preserved with carbon dioxide.

Cow's, ewe's, and goat's milk samples were treated with carbon dioxide gas until a pH of 6.1 was reached and stored at 4 degrees C to determine the resulting modifications in the mineral balance. The amounts of calcium and phosphorus dissolved during the acidification were similar in the three species. The acidification with CO2 produced the dissolution of phosphorus and magnesium in concentrations similar to those attained by acidification with lactic acid or hydrochloric acid. Still, the contents of soluble calcium and ionic calcium were higher with the CO2 treatment. The increase of ionic calcium due to the addition of CO2 could explain why milk subjected to such treatment is better suited for coagulation. Removal of added CO2 by shaking the milk for several hours at atmospheric pressure resulted in a higher concentration of ionic-calcium than was found in control milks to which no CO2 had been added. Thus the addition of CO2 improved milk's technological suitability for cheesemaking.

Isolation of prebiotic carbohydrates by supercritical fluid extraction. Scaling-up and economical feasibility.

Production of prebiotic carbohydrates at competitive prices is a challenge nowadays since the well-established production processes involve many purification steps which are labour intensive and require important amounts of reagents and products thus increasing prebiotic's price. Several processes have been studied in our laboratory involving the use of Supercritical Fluid Technology to fractionate and purify carbohydrate solid mixtures. Research carried out at laboratory scale using theoretical mixtures (lactose/lactulose and galactose/tagatose), commercially available carbohydrate mixtures and carbohydrate mixtures produced by enzymatic transglycosylation and isomerized with complexating reagents demonstrated that purification of prebiotic carbohydrates was technically possible by supercritical fluid extraction. In the present work, the process optimized at laboratory scale to fractionate carbohydrate mixtures produced by enzymatic transglycosylation has been scaled-up to an industrial level and its economic feasibility has been simulated employing AspenONE(®) V7.3 software to obtain consistent data supporting the interest of a potential investment for prebiotics production at large scale using supercritical fluids.

Preservation of raw milk with CO2. Sensory evaluation of heat-processed milks.

The effect of CO2 on the growth of psychrotrophic milk spoilage organisms was studied, both in raw fresh milk and in pure cultures of three species of Pseudomonas growing in sterilised milk. Changes of sensory properties of CO2-treated samples after heat treatment were also analysed. Inhibition of psychrotrophic growth at 7 degrees C in milk treated with CO2 to a pH 6.2 or 6.0 was impaired by a gradual reduction of the CO2 content during storage. Growth inhibition was considerably improved by pH adjustment at 24-h intervals. Sensory analysis showed significant differences between non-acidified and acidified samples after heat treatment at 75 degrees C for 20 s or 110 degrees C for 5 min. No sensory differences were found between non-acidified and acidified milks degassed before heat treatment.

Proteolysis during storage of UHT milk: differences between whole and skim milk.

Proteolysis during storage of UHT skim and whole milks processed by either direct or indirect systems has been studied. All the proteolysis indices determined (measurement of free amino groups, PAGE of caseins and PAGE and reversed-phase HPLC of the fraction soluble at pH 4.6) revealed greater proteolytic degradation during storage of skim milks compared with that of whole milks subjected to the same UHT treatments. Increased activities of both native milk proteinase and proteinases of bacterial origin were observed in skim UHT milks. The different behaviour of UHT skim and whole milks on storage would have to be taken into account in establishing the process conditions.

Comparative studies on "acid-stable, heat-stable polypeptides" of ovine, caprine and bovine milks.

"Acid-stable, heat-stable polypeptides" (previously called proteose-peptone) of three milk species have been studied. Analysis by polyacrylamide gel electrophoresis revealed that all fractions showed similar patterns. Only bands with the lowest mobilities showed the presence of carbohydrates when stained with Schiff's reagent. These bands had a lower mobility in ewe's and goat's milk than in cow's milk. Gas chromatography/mass spectrometry analysis showed the presence of fucose, mannose, glucose, galactose, glucosamine and galactosamine in the three fractions studied. Acid-stable, heat-stable polypeptides of bovine milk had the lowest carbohydrate content (4 mg/100 mg protein), whereas the highest content was found in ewe's milk (7.30 mg/100 mg protein) mainly as a result of the high galactose, mannose and glucosamine content. The sialic acid content was lower in ewe's and goat's milk (2.68 and 2.98 mg/100 mg protein respectively) than in bovine milk (2.06 mg/100 mg protein).

Effect of Storage Temperature on Lactulose and 5-hydroxymethyl-furfural Formation in UHT Milk.

The amounts of two heating-induced products were studied (lactulose and 5-hydroxymethyl-furfural) in UHT milk samples, directly and indirectly heat-treated, during a period of 90 d storage at five different temperatures. The results indicate an increase in 5-hydroxymethyl-furfural, as well as lactulose, taking place in the range of temperatures between 40-50°C.