Rat age-related benign prostate hyperplasia is concomitant with an increase in the secretion of low ramified α-glycosydic polysaccharides.

This work analysed the expression of prostate polysaccharides in rats with age-related benign prostatic hyperplasia (BPH) for a better understanding of the possible relationship between prostate polysaccharides secretion and BPH onset. For this, prostatic glands from 1 month-old, 3 months-old, 6 months-old and 12 months-old Sprague-Dawley rats were processed in order to identify their overall polysaccharide content. Additionally, serum testosterone was also determined. One-month old rats showed significantly (P < 0.05) lower testosterone levels (0.77 ng/mL±0.12 ng/mL) compared with the other groups, which showed no significant difference among them. PAS staining showed positive polysaccharides markings in both the prostatic lumen and inside of luminal prostatic cells in all groups. Semiquantitative analysis of intraluminal PAS showed that one month-old rats had significantly (P < 0.005) lower PAS intensity when compared with all other groups (100.0 ± 0.5, arbitrary units vs. 107.3 ± 0.6, arbitrary units in 3 months-old ones), whereas 12 months-old ones showed significantly (P < 0.005) higher values when compared with all other groups (133.6 ± 3.5, arbitrary units in 12 months-old rats vs. 108.6 ± 1.4, arbitrary units in 6 months-old ones). The PAS + content practically disappeared when tissues were pre-incubated with either α-amylase or amyloglucosidase, regardless of a previous incubation with proteinase K. Incubation of prostate extracts from 12 months-old rats for 2 h with α-amylase yielded a significantly higher amount of free glucose (1.47 nmol/mg protein±0.23 nmol/mg protein vs. 0.32 nmol/mg protein±0.01 nmol/mg protein in untreated extracts). Similar results were obtained when extracts were pre-incubated with amyloglucosidase. Contrarily, pre-incubation with N-glycosidase induced a significantly (P < 0.05), much lower increase of free glucose. Pre-treatment with proteinase K did not significantly modify these results, which indicate that BPH is related to an increase in the secretion of low ramified ductal α-glycosydic polysaccharides that were not protected against lysis by any type of protein protective core. These changes seem to not be related with concomitant variations in serum testosterone levels.

The onset of age-related benign prostatic hyperplasia is concomitant with increased serum and prostatic expression of VEGF in rats: Potential role of VEGF as a marker for early prostatic alterations.

The onset of age-related benign prostate hyperplasia (BPH) is linked with changes in the expression of specific prostatic chemokines. The aim of this work was to characterize those most relevant changes through the simultaneous analysis of 34 chemokines in both prostatic tissue and serum in rats at different ages with the aim to identify clinically workable parameters for the detection of early prostatic alterations. The study included 28 healthy Sprague-Dawley male rats that were distributed in four groups, 1 month-old (prepuberal; n = 7), 3 months-old (young; n = 7), 6 months-old (mature; n = 7) and 12 months-old (elder; n = 7). Chemokines were analyzed through a commercial mini-array system specially designed for rat tissues. Serum testosterone levels and prostatic histological status were also evaluated. Histological lesions indicative of BPH were detected in three mature rats and in all elder ones. Mini-arrays from prostatic tissue showed that young animals had an overall decreased expression of most of the analyzed chemokines when compared with prepuberal rats, with the exception of agrin, which showed a significant (P < 0.05) increase (100.0 ± 1.3, arbitrary units in prepuberal rats vs.148.2 ± 4.1, arbitrary units in young ones). Older animals showed further specific changes in 4 out 34 analyzed chemokines, namely agrin, platelet-derived growth factor (PDGF), tissue inhibitor of metalloproteinase-1 (TIMP-1) and vascular endothelial growth factor (VEGF). Additionally, elder rats showed the lowest intensity levels of agrin combined with the highest ones for PDGF, TIMP1 and VEGF when compared with all other groups. Finally, a significant increase of serum VEGF was detected in elder, BPH-affected rats when compared with young ones. Results indicated that the onset of both rat puberty and BPH would be related with specific changes in the prostatic expression of chemokines such as VEGF. Otherwise, the observed changes in serum VEGF levels could suggest the future possible utilization of serum VEGF levels to detect early pathological prostatic processes.

Effects of intrauterine devices on proteins in the uterine lavage fluid of mares.

Intrauterine devices block luteolysis in cyclic mares, but the underlying mechanism is unknown. To clarify the mechanisms, the protein profile of the endometrial secretome was analyzed using two-dimensional difference gel electrophoresis (2D-DIGE). Twenty-seven mares were classified according to whether they were inseminated (AI) or had an intrauterine device (IUD), a water-filled plastic sphere, inserted into the uterus on Day 3 after ovulation. Uterine lavage fluids were collected on Day 15 from pregnant inseminated mares (AI-P; n = 8), non-pregnant inseminated mares (AI-N; n = 4), and mares with IUD (n = 15). The IUD group was further divided into prolonged (IUD-P; n = 7) and normal luteal phase (IUD-N; n = 8) groups on the basis of ultrasound examinations, serum levels of progesterone and PGFM on Days 14 and 15, and COX-2 results on Day 15. Four mares from each group were selected for the 2D-DIGE analyses. Ten proteins had significantly different abundance among the groups, nine of the proteins were identified. Malate dehydrogenase 1, increased sodium tolerance 1, aldehyde dehydrogenase 1A1, prostaglandin reductase 1, albumin and hemoglobin were highest in pregnant mares; T-complex protein 1 was highest in non-pregnant mares; and annexin A1 and 6-phosphogluconolactonase were highest in IUD mares. The results suggest that the mechanism behind the intrauterine devices is likely related to inflammation.

Comorbilidad y síntomas digestivos asociados a depresión en pacientes mayores de 60 años / Gastrointestinal comorbidity and symptoms associated with depression in patients aged over 60 years

OBJETIVO: Conocer la prevalencia de enfermedades y síntomas digestivos asociados a la depresión. MATERIAL Y MÉTODOS: Estudio observacional transversal en atención primaria. Sujetos: todas las personas de 60 años o más con diagnóstico activo en 2017 de depresión del centro (N=2.312), comparados con los pacientes atendidos de igual edad sin depresión (N=11.049). Variables: edad, sexo, códigos de síntomas y diagnósticos de enfermedades crónicas digestivas. Fuente: historia clínica informatizada. Se ha calculado las prevalencias en ambos grupos, el odds ratio (OR) e intervalo de confianza (IC) del 95% para los principales diagnósticos y síntomas digestivos. RESULTADOS: El 38,0% de pacientes con depresión tiene alguna enfermedad digestiva crónica, el 64,7% algún síntoma y el 74,7% algún diagnóstico o síntoma digestivo. Los diagnósticos con OR significativo con relación a los pacientes sin depresión son: síndrome de intestino irritable OR: 2,0 (IC 95%: 1,51-2,64), anomalía congénita digestiva 1,83 (1,17-2,86), úlceras pépticas 1,75 (1,28-2,40), enfermedad diverticular 1,73 (1,52-1,97), colecistitis/colelitiasis 1,59 (1,36-1,86), hepatopatía 1,55 (1,32-1,82), hepatitis vírica 1,50 (1,12-2,02), enfermedades esofágicas 1,45 (1,24-1,69). Los síntomas con OR significativo son: anorexia 2,81 (1,75-4,50), náuseas/vómitos 2,19 (1,79-2,67), estreñimiento 1,96 (1,77-2,18), flatulencia 1,78 (1,48-2,14), disfagia 1,72 (1,30-2,28), dolor abdominal 1,69 (1,57-1,82), dispepsia 1,56 (1,30-1,87) y acidez 1,55 (1,26-1,92). CONCLUSIONES: La comorbilidad digestiva es muy frecuente en pacientes mayores de 60 años con depresión, hasta tres cuartos presentan enfermedades o síntomas digestivos. Las enfermedades más asociadas son el síndrome del intestino irritable, anomalías congénitas, otras úlceras pépticas y enfermedad diverticular. Los síntomas con mayor asociación son anorexia, náuseas/vómitos, estreñimiento, flatulencia, disfagia, dolor abdominal, dispepsia y acidez

OBJECTIVE: A study was carried out on the prevalence of gastrointestinal diagnoses and symptoms associated with depression. MATERIAL AND METHODS: A cross-sectional observational study was conducted in Primary Care in 2017. All patients aged 60 years or more with depression were included from the health care centre (N=2312), and were compared to the total patients that visited the centre from the same age group without depression (N=11049). The variables collected were age, gender and the codes for gastrointestinal symptoms and chronic conditions. These were obtained from the computerised medical files. The prevalence was calculated for both groups, and included the odds ratio (OR) and 95% confidence interval (CI). RESULTS: More than one-third (38.0%) of patients with depression have chronic gastrointestinal diseases, 64.7% have gastrointestinal symptoms, and 74.7% have either of them. The conditions with strongest associations are: irritable bowel syndrome OR: 2.00 (95% CI: 1.51-2.64), congenital anomaly 1.83 (1.17-2.86), other peptic ulcers 1.75 (1.28-2.40), diverticular disease 1.73 (1.52-1.97), cholecystitis/cholelithiasis 1.59 (1.36-1.86), liver disease 1.55 (1.32-1.82), viral hepatitis 1.50 (1.12-2.02), and oesophageal diseases 1.45 (1.24-1.69). Symptoms with a statistically significant OR were: anorexia 2.81 (1.75-4.50), nausea/vomiting 2.19 (1.79-2.67), constipation 1.96 (1.77-2.18), flatulence 1.78 (1.48-2.14), dysphagia 1,72 (1.30-2.28), abdominal pain 1.69 (1.57-1.82), dyspepsia 1.56 (1.30-1.87), and heartburn 1.55 (1.26-1.92). CONCLUSIONS: Gastrointestinal comorbidity is very common in patients over 60 years-old with depression. Three-quarters of the patients have chronic gastrointestinal diseases or symptoms. The conditions with strongest associations are irritable bowel syndrome, congenital anomalies, other peptic ulcers, and diverticular disease. The symptoms with strongest associations are anorexia, nausea/vomiting, constipation, flatulence, dysphagia, abdominal pain, dyspepsia, and heartburn

[Gastrointestinal comorbidity and symptoms associated with depression in patients aged over 60 years]. / Comorbilidad y síntomas digestivos asociados a depresión en pacientes mayores de 60 años.

OBJECTIVE: A study was carried out on the prevalence of gastrointestinal diagnoses and symptoms associated with depression. MATERIAL AND METHODS: A cross-sectional observational study was conducted in Primary Care in 2017. All patients aged 60 years or more with depression were included from the health care centre (N=2312), and were compared to the total patients that visited the centre from the same age group without depression (N=11049). The variables collected were age, gender and the codes for gastrointestinal symptoms and chronic conditions. These were obtained from the computerised medical files. The prevalence was calculated for both groups, and included the odds ratio (OR) and 95% confidence interval (CI). RESULTS: More than one-third (38.0%) of patients with depression have chronic gastrointestinal diseases, 64.7% have gastrointestinal symptoms, and 74.7% have either of them. The conditions with strongest associations are: irritable bowel syndrome OR: 2.00 (95% CI: 1.51-2.64), congenital anomaly 1.83 (1.17-2.86), other peptic ulcers 1.75 (1.28-2.40), diverticular disease 1.73 (1.52-1.97), cholecystitis/cholelithiasis 1.59 (1.36-1.86), liver disease 1.55 (1.32-1.82), viral hepatitis 1.50 (1.12-2.02), and oesophageal diseases 1.45 (1.24-1.69). Symptoms with a statistically significant OR were: anorexia 2.81 (1.75-4.50), nausea/vomiting 2.19 (1.79-2.67), constipation 1.96 (1.77-2.18), flatulence 1.78 (1.48-2.14), dysphagia 1,72 (1.30-2.28), abdominal pain 1.69 (1.57-1.82), dyspepsia 1.56 (1.30-1.87), and heartburn 1.55 (1.26-1.92). CONCLUSIONS: Gastrointestinal comorbidity is very common in patients over 60 years-old with depression. Three-quarters of the patients have chronic gastrointestinal diseases or symptoms. The conditions with strongest associations are irritable bowel syndrome, congenital anomalies, other peptic ulcers, and diverticular disease. The symptoms with strongest associations are anorexia, nausea/vomiting, constipation, flatulence, dysphagia, abdominal pain, dyspepsia, and heartburn.

Efficacy of an information system addressed to nursing staff for diminishing contaminated blood cultures: a blind clinical trial.

OBJECTIVE: Evaluate the efficacy of an information system addressed to nursing staff to lower the blood culture contamination rate. METHODS: A blind clinical trial was conducted at Internal Medicine and Emergency Departments during 2011. After following a reeducation program in BC extraction, participants were randomly selected in a 1:1 ratio. Every participant of the experimental group was informed of each worker's individual performance; whereas the control group was only informed of the global results. RESULTS: A total of 977 blood extractions were performed in 12 months. Blood culture contamination rate was 7.5%. This rate was higher in the Emergency Department than in Internal Medicine (10% vs. 3.8%; p=0.001). Factors associated with the higher risk of contamination were, in the univariate analysis, the extraction through a recently implanted blood route and the time of professional experience, while those associated with a lower risk were the extraction in Internal Medicine and through a butterfly needle. On multivariate analysis, extraction through a recently placed access was an independent risk factor for an increased contamination rate (OR 2.29; 95%CI 1.18-4.44, p=0.014), while individual information about the blood culture results (OR 0.11; 95%CI 0.023-0.57; p=0.008), and more than 9 years of professional experience were asso-ciated with fewer contaminations (OR 0.30; 95%CI 0.12-0.77; p=0.012). In the intervention group the contamination rate diminished by a 26 %. CONCLUSIONS: Drawing blood cultures through a recently taken peripheral venous access increased their risk of contamination. The intervention informing the nurse staff of the contamination rate is effective to decrease it.

Relationship of aquaporins 3 (AQP3), 7 (AQP7), and 11 (AQP11) with boar sperm resilience to withstand freeze-thawing procedures.

Cryopreservation is the most suitable method to preserve boar spermatozoa over long-term storage. However, freeze-thawing protocols inflict extensive damage to sperm cells, reducing their viability and compromising their fertilizing ability. In addition, high individual variability is known to exist between boar ejaculates, which may be classified as of good (GFE) or poor (PFE) freezability. While conventional spermiogram parameters fail to predict sperm cryotolerance in fresh spermatozoa, high levels of certain proteins, also known as freezability markers, have been found to be related to the sperm resilience to withstand freeze-thawing procedures. In this context, the hypothesis of this study was that aquaporins AQP3, AQP7, and AQP11 could be linked to boar sperm cryotolerance. Twenty-nine ejaculates were evaluated and subsequently classified as GFE or PFE based upon their sperm viability and motility at post-thawing. Fourteen ejaculates resulted to be GFE, whereas the other fifteen were found to be PFE. Relative abundances of AQP3, AQP7, and AQP11 and their localization patterns were evaluated in all fresh and frozen-thawed ejaculates through immunoblotting and immunocytochemistry. Prior to cryopreservation, relative amounts of AQP3 and AQP7 were found to be significantly (p < 0.05) higher in GFE than in PFE. In contrast, no significant differences (p > 0.05) between freezability groups were found for AQP11, despite GFE tending to present higher levels of this protein. The localization of AQP7, but not that of AQP3 or AQP11, was observed to be affected by cryopreservation procedures. In conclusion, these results suggest that AQP3 and AQP7 are related to boar sperm cryotolerance and may be used as freezability markers.

Effects of Intrauterine Devices in Mares: A Histomorphological and Immunohistochemical Evaluation of the Endometrium.

Oestrous suppression by intrauterine devices (IUDs) is caused by prolongation of luteal function, but the biological mechanism is unknown. The aim of the study was to investigate mechanisms which could explain the action of IUDs. Thirty mares were age-matched and either inseminated (AI, n = 15) or fitted with an IUD (IUD, n = 15) and subsequently divided into four groups: AI-P, pregnant (n = 8); AI-N, non-pregnant (n = 7); IUD-P, prolonged luteal phase (n = 7); and IUD-N, normal luteal phase (n = 8). The median ages were 5.5 and 7 years in AI-P and IUD-P groups and 14 and 11 years in AI-N and IUD-N groups, respectively. On Day 15 after ovulation, an endometrial biopsy was obtained to study histomorphological and immunohistochemical expression patterns of uterine proteins (uteroferrin, UF; uterocalin, UC; uteroglobin, UG), oestrogen and progesterone receptors (ER, PR), proliferation marker Ki-67 and content of inflammatory cells. Expression of UF was higher in IUD mares; the difference between pregnant and IUD-P mares was significant. Mares exhibiting a prolonged luteal phase (AI-P, IUD-P) showed only mild angiosclerosis and lower expression of both ER and PR than mares with a normal luteal phase (AI-N, IUD-N). No significant differences were detected in the numbers of inflammatory cells, with the exception of macrophages, which were more numerous in AI-P than AI-N mares. Although inflammatory cells were not detected in IUD mares, increased UF levels may indicate chronic inflammation. Young age and normality of the endometrial blood vessels may improve the efficacy of IUDs.

Doppler evaluation of the effects of propofol, etomidate and alphaxalone on fetoplacental circulation hemodynamics in the pregnant ewe.

INTRODUCTION: Umbilical artery (UA) hemodynamics reflect blood flow and vascular resistance in the placental circulation. We examined non-invasively the hemodynamic effects of propofol, etomidate, and alphaxalone on the placental circulation of a sheep model by means of UA Doppler ultrasonography. METHODS: Eleven sheep fetuses were examined at 90-109 days of gestation. UA Doppler ultrasound was performed before and after administration of a single intravenous bolus of propofol, etomidate, or alphaxalone. UA Doppler velocities (peak systolic velocity, end diastolic velocity, and mean velocity), vascular indices (pulsatility index, resistance index, and S/D ratio), blood flow, and fetal heart rate were recorded during the experimental period and UA Doppler waveforms were characterized. RESULTS: A laminar, parabolic, low resistance flow was observed in the UA of the sheep fetuses. No statistically significant changes were observed in the UA Doppler waveforms or in the UA Doppler hemodynamics after anesthesia induction. DISCUSSION: Changes in placental vascular resistance may alter the corresponding UA Doppler waveforms. When resistance in the fetal placenta increases, blood flow in the UA becomes more pulsatile. In the present study, umbilical arteries showed a parabolic flow with low resistance in all cases, as it occurs in normal human pregnancy. The administration of these anesthetics did not cause abnormalities in the normal UA Doppler pattern, inducing no changes in the resistance of the placenta in any case. CONCLUSION: These results suggest that intravenous anesthetic induction with propofol, etomidate, or alphaxalone does not cause significant detrimental effects on the placental circulation of the pregnant ewe.

Effects of ovariohysterectomy on intra-abdominal pressure and abdominal perfusion pressure in cats.

Intra-abdominal pressure (IAP) and abdominal perfusion pressure (APP) have shown clinical relevance in monitoring critically ill human beings submitted to abdominal surgery. Only a few studies have been performed in veterinary medicine. The aim of this study was to assess how pregnancy and abdominal surgery may affect IAP and APP in healthy cats. For this purpose, pregnant (n=10) and non-pregnant (n=11) queens undergoing elective spaying, and tomcats (n=20, used as controls) presented for neutering by scrotal orchidectomy were included in the study. IAP, mean arterial blood pressure (MAP), APP, heart rate and rectal temperature (RT) were determined before, immediately after, and four hours after surgery. IAP increased significantly immediately after abdominal surgery in both female groups when compared with baseline (P<0.05) and male (P<0.05) values, and returned to initial perioperative readings four hours after surgery. Tomcats and pregnant females (P<0.05) showed an increase in MAP and APP immediately after surgery decreasing back to initial perioperative values four hours later. A significant decrease in RT was appreciated immediately after laparotomy in both pregnant and non-pregnant queens. IAP was affected by abdominal surgery in this study, due likely to factors, such as postoperative pain and hypothermia. Pregnancy did not seem to affect IAP in this population of cats, possibly due to subjects being in early stages of pregnancy.

Modulation of the biochemical composition of amniotic and allantoic fluids as a control mechanism of feline foetal development.

Amniotic fluid exerts a protective function and is an essential component for foetal development and maturation during pregnancy. However, little is known about the exact physiological functions of foetal fluids in this process as well as their biochemical composition in cats. In the present study, the biochemical composition of amniotic and allantoic fluids and maternal serum in pregnant queens was compared after performing an ovariohysterectomy. Fifteen queens were included in the study and distributed in four different groups, D(30), D(40), D(50) and D(60), according to their gestational age. Foetal fluids showed thoroughly greater concentrations of dissociate and total bilirubin, bile acids and gamma-glutamyl transferase than those of maternal serum, whereas albumin, total protein, alanine-transferase, creatine-kinase, amylase, lipase, triglycerides, cholesterol, HDL-cholesterol, and LDL-cholesterol were significantly lower, as compared to maternal serum. Other parameters like alkaline phosphatase, uric acid, creatinine, and electrolytes showed significant differences at specific stages of pregnancy, when compared to maternal serum. Lactate and cortisol significantly increased at the end of the pregnancy in foetal fluids, when compared with maternal serum. No significant differences between foetal fluids and maternal serum were observed for aspartate aminotransferase, lactate dehydrogenase, urea, phosphorus and glucose. According to our results, foetal fluids composition is not a result of simple filtration from maternal blood, the fetus being an active element involved in the production of the same and reflecting organ development and maturation.

Phenolic compounds and sensorial characterization of wines aged with alternative to barrel products made of Spanish oak wood (Quercus pyrenaica Willd.).

Wood of Quercus pyrenaica has suitable properties for the wine ageing process. However, the forest available for the barrel making from this particular type of tree is very limited. Nevertheless, it is highly advisable to use this kind of wood in order to manufacture alternative oak products. This study presents the results of ageing the same red wine using different pieces of wood (chips and staves) of Spanish oak (Q. pyrenaica), American oak (Quercus alba) and French oak (Quercus petraea) in conjunction with small, controlled amounts of oxygen. In addition, the phenolic parameters, colour and sensory analysis point out that wines aged with Q. pyrenaica pieces have similar enological characteristics to those aged with American or French oak pieces of wood (chips and staves). Furthermore, the total oxygen consumed and its relation with sensory properties also has been studied in this article in order to know how the oxygen behaves in these processes. Besides, it is going to put forward the fact that chips and staves from Q. pyrenaica oak are suitable for the ageing of red wines and better considered than American or French ones, showing higher aromatic intensity, complexity, woody, balsamic and cocoa. Finally, the tasters valued highly the wines with staves, pointing out its flavour and roundness in mouth.

Determination of low-molecular mass phenols in red wines: the influence of chips, staves and micro-oxygenation aging tank.

The aim of this study is to assess the use of polymeric cartridges with diode array detection to extract the main low-molecular mass phenolic compounds from red wine samples. This method expanded and improved permitted to quantify 20 compounds in red wines made and aged under different conditions. These conditions included aging in barrels and in tanks with chips or staves as well as micro-oxygenation. The study of wine development during its wood storage period and the compounds transferred from the wood to the wine during this period provide useful important information concerning the impact of wine aging methods on the chemical makeup of the resulting wines. The main compounds that differentiated wine aging systems were identified to be 5-hydroxymethyl-2 furaldehyde, gallic acid, p-hydroxybenzoic aldehyde, syringic acid and sinapic acid.

Partial foetal retention following aglepristone treatment in a bitch.

This short communication describes the case of partial foetal retention in an 18-month-old female French bulldog following induction of abortion owing to an undesired mating. Abortion was induced with aglepristone administered in two consecutive protocols of a dual injection 1 day apart. After failure of the first treatment to achieve abortion, 15 days later, a second treatment was administered. Delivering of aborted foetus occurred 2 days after the last administration. Five weeks after the abortion, the female showed a weak haemorrhagic vaginal discharge. On ultrasound examination, the presence of uterine wall distension as well as a puppy skull inside the uterus was observed. This clinical case makes clear that although aglepristone is a very reliable drug, follow-up of the female during treatment and in the immediate post-partum is necessary to ensure a good outcome.

Protein composition of seminal plasma in fractionated stallion ejaculates.

Seminal plasma (SP) contains several types of compounds derived from the epididymides and accessory glands. The aim of this study was to examine the protein composition of different ejaculate fractions. Trial I: fractionated ejaculates were collected from two normal and two subfertile stallions. Samples containing pre-sperm fluid and the first sperm-rich jets (HIGH-1), the main sperm-rich portion (HIGH-2), the jets with low sperm concentrations (LOW), and a combined whole-ejaculate (WE) sample was centrifuged, and the SP was filtered and frozen. A part of each SP sample was stored (5°C, 24 h) with spermatozoa from HIGH-2 and skim milk extender. Sperm motility was evaluated after storage in extender mixed with the stallion's own SP or SP from one of the other stallions (sperm from a normal stallion stored in SP from a subfertile stallion and vice versa). Protein composition was analysed using reverse-phase liquid chromatography (RP-HPLC), N-terminal sequencing and mass spectrometry. The area-under-the-curve (AUC) was used for quantitative comparison of proteins within fractions. Trial II: semen samples were collected from seven stallions. Fractions with the highest (HIGH) and lowest (LOW) sperm concentrations and WE samples were examined using SDS-PAGE and densitometry. No significant differences emerged between fractions in the AUC-values of the Horse Seminal Protein-1 (HSP-1) and HSP-2 peaks, or the peak containing HSP-3 and HSP-4 (HSP-3/4). Levels of HSP-1, HSP-2 and HSP-3/4 were not significantly correlated with total sperm motility, progressive sperm motility or average path velocity after storage. Significant differences between ejaculate fractions in the amount of different protein groups present in SP were not found in Trial I; but in Trial II, the proteins in the 60-70 kDa range were more abundant in LOW than in HIGH and WE, indicating that this band contained proteins derived mainly from the seminal vesicles, which produce most of the SP in LOW.

Effect of size, seasoning and toasting in the volatile compounds in toasted oak wood and in a red wine treated with them.

The increasing demand for wood for barrel-making in addition to the rapid extension of alternative aging system, have led to looking into the possibility of utilizing Spanish oak. Quercus pyrenaica is the species that predominates in Spain, and the chemical composition of its heartwood (ellagitannins, low molecular weight phenolic and volatile compounds) and its incidence in characteristics of wine are similar to that of other species that are of recognized oenological quality for barrel-making, showing only quantitative differences with respect to French (Quercus petraea) and American (Quercus alba) species. However, at present, the quantity of good quality wood that we can obtain from the Q. pyrenaica Spanish forest is limited. Hence, in the short term, and considering the high chemical oenological quality of Q. pyrenaica wood, we propose the utilizing of chips, segments, staves, and other oak alternatives for wine aging, which would be obtained from wooden remnants from barrel-making as well as from trees with small diameters or physical defects which would normally be inappropriate for cooperage. With regards to the latter idea, studies on special chip-making processes, and other oak wood pieces are being carried out, especially focused on reducing seasoning time, and to toasting optimization as a function of wood piece size, in addition to its behaviour when incorporated into the different alternative aging systems. We present in this study the effect of seasoning way (traditional or unconventional) on volatile composition of Q. pyrenaica chips and staves at three toasting levels (light, medium and heavy), and the evolution of the wood-released aromatic composition of a Spanish artificially aged wine, using these alternative products. The wines showed in general small differences in their oak-derived characteristics, which were more related to the wood piece size and the toasting intensity than to the seasoning way, and they could be linked with the concentrations of compounds as eugenol, furfural, and the cis and trans isomers of beta-methyl-gamma-octolactone.

Dissolved oxygen distribution during micro-oxygenation. Determination of representative measurement points in hydroalcoholic solution and wines.

Red wine tank aging is monitored by organoleptic analysis, therefore, it is necessary to use an objective parameter representing the process. Among the possible parameters to be checked, it stands out the knowledge of dissolved oxygen because it offers the possibility of anticipating undesirable situations that bring about too much oxidation. Dissolved oxygen measurement, with non-intrusive luminescent technology is becoming an effective alternative. Uncertainty arises when trying to choose the measuring point able to represent the entire tank since previous works have considered the existence of gradients throughout the volume of the treated wine. This paper shows the results obtained from the study of the existence and the quantification of gradients of the dissolved oxygen in a 15% hydroalcoholic solution during the micro-oxygenation process. Different measuring point placements are studied and the solutions to monitor the process by controlling a representative point are set out. A successful monitoring of a red wine tank aging with alternative oak products and adaptative micro-oxygenation has proved that an objective control of the process is, indeed, possible.

Ultrarapid quantitation of maize proteins by perfusion and monolithic reversed-phase high-performance liquid chromatography.

The main objective of this study was to develop a new methodology alternative to the classical Kjeldahl analysis for determining maize proteins in maize products and seeds. For that purpose, two different chromatographic methodologies using perfusion and monolithic stationary phases, both enabling rapid separations of maize proteins, were investigated. Due to the difficulty to find suitable standards for this type of analysis, three different maize products were initially tested as proteins standards: zein F4000, corn gluten meal, and maize flour. Different figures of merit (i.e., linearity, correlation coefficient, precision, limits of detection and quantitation), as well as the presence of matrix inferences, were investigated. The results obtained for the different chromatographic stationary phases and protein standards were compared in order to select the most suitable analytical conditions. Despite both perfusion and monolithic methodologies resulting, in general, as appropriate for the quantitation of maize proteins, the highest reduction of analysis time and lowest detection and determination limits provided by perfusion methodology enabled to select this one as the method of choice for the quantitation of maize proteins. Regarding the different protein standards studied in this work, in general the best results were obtained using the zein standard. Compared to Kjeldahl methodology, perfusion chromatography yields total protein contents in shorter analysis time while enabling the separation of the different kinds of proteins. Due to the high diversity and complexity of industrial maize products, the proposed chromatographic method could be a very useful tool for their routine analysis.

Measurement of dissolved oxygen during red wines tank aging with chips and micro-oxygenation.

Nowadays, micro-oxygenation is a very important technique used in aging wines in order to improve their characteristics. The techniques of wine tank aging imply the use of small doses of oxygen and the addition of wood pieces of oak to the wine. Considering the low dissolved oxygen (DO) levels used by micro-oxygenation technique it is necessary to choose the appropriate measurement principle to apply the precise oxygen dosage in wine at any time, in order to assure its correct assimilation. This knowledge will allow the oenologist to control and run the wine aging correctly. This work is a thorough revision of DO measurement main technologies applied to oenology. It describes the strengths and weaknesses of each of them, and draws a comparison of their workings in wine measurement. Both, the traditional systems by electrochemical probes, and the newest photoluminescence-based probes have been used. These probes adapted to red wines ageing study are then compared. This paper also details the first results of the dissolved oxygen content evolution in red wines during a traditional and alternative tank aging. Samples have been treated by three different ageing systems: oak barrels, stainless-steel tanks with small oak wood pieces (chips) and with bigger oak pieces (staves) with low micro-oxygenation levels. French and American oak barrels manufactured by the same cooperage have been used.

Aging markers from bottled red wine aged with chips, staves and barrels.

This paper shows the results of the experiments carried on Spanish red wines aged through alternative as well as traditional oak barrel systems, i.e., chips stainless steel tanks and staves stainless steel tanks, at the same time and under the same conditions. Wine aging through wood pieces is not accepted as an enological practice by the European Union and, in fact, it is considered a fraud. It is, then, of a great interest to identify those parameters able to establish the aging technique that had been used in a wine ready to be consumed. With this idea in mind, the development of the same wine has been studied while aging for 3 years in three different systems. During the first six aging months wines stored with staves obtained characteristics that were halfway between wines treated with chips and those aged in barrels. However, as wood contact period length increased so did the differences between wines stored in traditional and alternative systems (either with staves or chips). These differences grew during the bottling period, so that after a 2-year bottling period wines from the three systems became different enough to tell them apart. Discriminant analysis of the variables studied made it possible to establish these differences. The most meaningful variables were yellow colour component, anthocyanins (cyanidin-3-glucoside, vitisin A and sum of pcoumaryl derivates), vanillin acid, protocachuic aldehyde and epicatechin.