In vivo evidence that local cortisol production increases in the preovulatory follicle of the cow

The aim of the present in vivo study was to monitor real-time fluctuations of cortisol (Cr) in the wall of preovulatory follicles using a microdialysis system (MDS) implanted in the theca layer as well as changes in ovarian venous plasma (OVP) and jugular venous plasma (JVP). Seven cows were superovulated using FSH and prostaglandin F2alpha injections. Dialysis capillary membranes were surgically implanted into the theca layer of mature follicles and connected to a microdialysis system. Fractions of the perfusates were collected from Day -1 (Day 0=LH surge) to Day 3. No difference in the concentrations of Cr between JVP and OVP was detected throughout the experiment. Circulating concentrations of Cr ranged from 20 to 35 ng/ml 8 h after surgery in ovulatory and anovulatory cows. In five ovulatory cows, the Cr concentration decreased to basal levels (<10 ng/ml) between 12 and 24 h after surgery, however, two anovulatory cows retained high Cr levels (>10 ng/ml) up to 42 h after surgery. There was a clear increase in the local concentration of Cr from 13.3+/-2.1 pg/ml at -24 h to 27.5+/-1.7 pg/ml at 0 h (peak of the LH surge) within the wall of ovulatory follicles. This increase was not detected in anovulatory follicles. This transient increase in Cr occurred only in the follicle wall, but not in the OVP or JVP, indicating that the presence of a local regulatory mechanism for Cr production/conversion in ovulatory follicles, and this mechanism may modulate the inflammatory-like reaction induced by LH surge in the follicle wall. The present results demonstrate that the glucocorticoid environment in the follicular wall adjusts at the local level in bovine ovulatory follicles. This mechanism may protect follicles from the adverse effects of glucocorticoid, and it may prevent excess inflammatory reactions associated with ovulation by temporarily increasing local concentrations of glucocorticoid, thus forming an integral part of the regulatory mechanism in ovarian physiology.     

Cardiopulmonary effects of fentanyl in conscious dogs and dogs sedated with a continuous rate infusion of medetomidine

OBJECTIVE: To determine the hemodynamic consequences of the coadministration of a continuous rate infusion (CRI) of medetomidine with a fentanyl bolus in dogs. ANIMALS: 12 healthy sexually intact male dogs weighing 30.3 -/+ 4.2 kg (mean +/- SD). PROCEDURE: Dogs received either fentanyl alone (15.0 microg/kg, i.v. bolus) or the same dose of fentanyl during an 11-hour CRI of medetomidine (1.5 microg/kg/h, i.v.). Prior to drug administration, dogs were instrumented for measurement of cardiac output, left atrial pressure, and systemic arterial blood pressures. Additionally, blood samples were collected from the pulmonary artery and left atrium for blood gas analysis. RESULTS: Medetomidine infusion reduced the cardiac index, heart rate, and O2, delivery while increasing left atrial pressure. Subsequent fentanyl administration further decreased the cardiac index. The Pao2 was not significantly different between the 2 treatment groups; however, fentanyl transiently decreased Pao2 from baseline values in dogs receiving a CRI of medetomidine. CONCLUSIONS AND CLINICAL RELEVANCE: Because of the prolonged hemodynamic changes associated with the CRI of medetomidine, its safety should be further evaluated before being clinically implemented in dogs.     

Effects of acepromazine and butorphanol on tiletamine-zolazepam anesthesia in llamas

OBJECTIVE: To evaluate sedative, antinociceptive, and physiologic effects of acepromazine and butorphanol during tiletamine-zolazepam (TZ) anesthesia in llamas. ANIMALS: 5 young adult llamas. PROCEDURES: Llamas received each of 5 treatments IM (1-week intervals): A (acepromazine, 0.05 mg/kg), B1 (butorphanol, 0.1 mg/kg), AB (acepromazine, 0.05 mg/kg, and butorphanol, 0.1 mg/kg), B2 (butorphanol, 0.2 mg/kg), or C (saline [0.9% NaCl] solution). Sedation was evaluated during a 30-minute period prior to anesthesia with TZ (2 mg/kg, IM). Anesthesia and recovery characteristics and selected cardiorespiratory variables were recorded at intervals. Antinociception was assessed via a toe-clamp technique. RESULTS: Sedation was not evident following any treatment. Times to sternal and lateral recumbency did not differ among treatments. Duration of lateral recumbency was significantly longer for treatment AB than for treatment C. Duration of antinociception was significantly longer for treatments A and AB, compared with treatment C, and longer for treatment AB, compared with treatment B2. Treatment B1 resulted in a significant decrease in respiratory rate, compared with treatment C. Compared with treatment C, diastolic and mean blood pressures were lower after treatment A. Heart rate was increased with treatment A, compared with treatment B1 or treatment C. Although severe hypoxemia developed in llamas anesthetized with TZ alone and with each treatment-TZ combination, hemoglobin saturation remained high and the hypoxemia was not considered clinically important. CONCLUSIONS AND CLINICAL RELEVANCE: Sedation or changes in heart and respiratory rates were not detected with any treatment before administration of TZ. Acepromazine alone and acepromazine with butorphanol (0.1 mg/kg) prolonged the duration of antinociception in TZ-treated llamas.     

Determination of 24 pesticide residues in fortified wines by solid-phase microextraction and gas chromatography-tandem mass spectrometry

The present work describes a solid-phase microextraction (SPME) gas chromatography-tandem mass spectrometry (MS/MS) method to quantify 24 pesticides in fortified white wine and fortified red wine. In this study "fortified wine" refers to a wine in which fermentation is arrested before completion by alcohol distillate addition, allowing sugar and alcoholic contents to be higher (around 80-100 g/L total sugars and 19-22% alcohol strength (v/v)). The analytical method showed good linearity, presenting correlation coefficients (R(2)) ≥ 0.989 for all compounds. Limits of detection (LOD) and quantitation (LOQ) in the ranges of 0.05-72.35 and 0.16-219.23 μg/L, respectively, were obtained. LOQs are below the maximum residue levels (MRL) set by European Regulation for grapes. The proposed method was applied to 17 commercial fortified wines. The analyzed pesticides were not detected in the wines tested.     

Metabolic profiling of ‘Conference’ pears under low oxygen stress

Pears (Pyrus communis L. cv. Conference) may develop core breakdown when stored under low oxygen or elevated carbon dioxide conditions. This physiological disorder is characterized by the development of brown spots due to oxidation of phenolic compounds, and eventually, cavities in the center of the fruit. Based on metabolic profiling of brown and sound tissue using GC-EI-TOF-MS, the hypothesis that this disorder is due to an imbalance between oxidative and reductive processes at the cellular level was investigated. Brown tissue was clearly characterized by a distinctive pattern in changes which included a decrease of malic acid and an increase in fumaric acid and gamma aminobutyric acid (GABA), which indicated a reduced metabolic activity at the level of the Krebs cycle and a putative block of the GABA shunt pathway. Increased gluconic acid concentration might be related to ascorbic acid degradation due to insufficient reducing equivalents or to an impaired pentose phosphate pathway. For the first time, GABA and gluconic acid have been shown to be metabolic markers for core breakdown. The concentrations of other compounds which are believed to be related to hypoxic stress response such as trehalose and putrescine were also considerably higher in brown tissue than in sound tissue. The concentration of some sugars which are typically found in xyloglucans also increased during brown development, possibly indicating cell wall breakdown due to enzymatic processes or chemical reactions of hydroxyl radicals.     

Inhibition of enzymatic browning of chlorogenic acid by sulfur-containing compounds

The antibrowning activity of sodium hydrogen sulfite (NaHSO(3)) was compared to that of other sulfur-containing compounds. Inhibition of enzymatic browning was investigated using a model browning system consisting of mushroom tyrosinase and chlorogenic acid (5-CQA). Development of brown color (spectral analysis), oxygen consumption, and reaction product formation (RP-UHPLC-PDA-MS) were monitored in time. It was found that the compounds showing antibrowning activity either prevented browning by forming colorless addition products with o-quinones of 5-CQA (NaHSO(3), cysteine, and glutathione) or inhibiting the enzymatic activity of tyrosinase (NaHSO(3) and dithiothreitol). NaHSO(3) was different from the other sulfur-containing compounds investigated, because it showed a dual inhibitory effect on browning. Initial browning was prevented by trapping the o-quinones formed in colorless addition products (sulfochlorogenic acid), while at the same time, tyrosinase activity was inhibited in a time-dependent way, as shown by pre-incubation experiments of tyrosinase with NaHSO(3). Furthermore, it was demonstrated that sulfochlorogenic and cysteinylchlorogenic acids were not inhibitors of mushroom tyrosinase.     

Stand density and fertilization effects on aboveground allocation patterns and stem form of Pinus sylvestris in young stands

The increasing demand for forest biomass for energy generation could be partially met by growing denser stands and use of fertilizer. Before this is done at large scale, more knowledge of the effects of stand density and fertilization on aboveground allocation patterns and stem form is needed. Therefore, effects of pre-commercial thinning (PCT) to 3000 stems ha^?1, an unthinned dense control (C), and PCT combined with two levels of fertilization (100 kg ha^?1 of nitrogen applied either during the establishment of the field experiment (F1) or annually (F2)) were examined in 23- to 26-year-old Scots pine (Pinus sylvestris L.) stands six years after the establishment of the field experiment. In total, 114 sample trees were harvested using destructive biomass sampling. The growth allocation and stem form of trees with diameter at breast height (DBH; 1.3 m height) >5.0 cm were not affected by either the PCT or fertilization. Small trees (DBH < 5 cm) in denser, unthinned control plots had more slender stems (lower DBH/height ratios) and allocated less growth to branches and foliage than trees in PCT plots. Fertilization had little effect on the stem form and growth allocation of the smallest trees. Therefore, effects of stem density and fertilization on stem form and growth allocation to foliage were only found for small suppressed trees, and the treatments had very little influence on dominant and codominant trees.