Red propolis as an additive for preweaned dairy calves: Effect on growth performance, health, and selected blood parameters.

Propolis is a natural product produced by bees and sold commercially. It is well known for its antioxidant and anti-inflammatory effects and the ability to improve immune system functions in humans and animals. Many of its positive effects can contribute positively to animal productivity. The aim of this study was to evaluate possible beneficial effects of red propolis supplementation on growth performance, metabolism, and health status of dairy calves during the preweaning phase. Thirty-two newborn calves were individually housed, with free access to water and concentrate, and fed 6 L/d of whole milk. Animals were distributed in 2 treatments: (1) control and (2) supplementation with 4 mL/d of red propolis ethanolic extract (30%; EEP) in whole milk. Feed consumption and fecal and health scores were monitored daily from 1 d of age to 56 d. Calves were weighed and measured weekly. Blood samples were collected weekly, 2 h after morning milk feeding, for determination of glucose, total serum protein, urea, lactate, ß-hydroxybutyrate, total iron binding capacity, and total leukocytes (white blood cells) and erythrocytes (red blood cells). Starter feed intake, daily weight gain, and body measurements were not affected by the EEP supplementation. Blood parameters were also not affected, with the exception for the red blood cell counts, which were lower in animals supplemented with EEP. Supplementation with EEP significantly decreased fecal score, days with diarrhea, and veterinary treatments of diarrhea. These results indicate that red propolis supplementation has potential to improve calf health and reduce the incidence of diarrhea and, as a consequence, the use of antibiotics in calf rearing systems.

Comparison and characterization of volatile compounds as markers of oils stability during frying by HS-SPME-GC/MS and Chemometric analysis.

The formation and emission of volatile compounds, including the aldehydes and some toxic compounds of oil samples, ROPO pure (100%) and the blended ROPO/RCO (80-20%), were carried out during deep frying at 180°C. The volatile profile of both oil samples was evaluated by an optimized HS-SPME-GC/MS method, before and after 20, 40 and 60 successive sessions of deep-frying. Actually, from 100 detected compounds, aldehydes were found to be the main group formed. In addition, the oil degradation under thermal treatment regarding the volatile compounds were evaluated and compared. Consequently, the blended ROPO/RCO revealed fewer formations of unsaturated aldehydes, including toxic ones, such as acrolein, and showed a greater stability against oxidative thermal degradation compared to ROPO pure.

Thalidomide reduces mechanical hyperalgesia and depressive-like behavior induced by peripheral nerve crush in mice.

BACKGROUND: It has been shown that chronic pain is able to induce depressive disorders in humans, in part, due to peripheral inflammation that reaches the central nervous system. However, the mechanisms involved remain to be established. The purpose of this study was to investigate whether sciatic nerve crush could produce depression-like behaviors, in addition to pain-related behaviors, in mice. Once confirmed, this model was used to investigate tumor necrosis factor-α (TNF-α) as a key mediator involved in the pathophysiology of both pain and depression. EXPERIMENTAL APPROACH: Male Swiss mice were divided into three groups, naïve, sham and operated. In the operated group, the sciatic nerve was crushed. Following surgery, animals from the operated group were treated daily by oral gavage (p.o.) with saline (10 ml/kg), fluoxetine (20 mg/kg) or thalidomide (10 mg/kg) for 15 days. Mechanical hyperalgesia was evaluated every 3 days by von Frey filaments and depressive-like behavior was assessed at the end of day 15, using the tail suspension test (TST) and the forced swimming test (FST). Then, samples from the prefrontal cortex, hippocampus and sciatic nerve were processed to measure TNF-α levels by enzyme-linked immunosorbent assay (ELISA). RESULTS: Crush caused significant mechanical hyperalgesia and depressive-like behaviors and increased TNF-α levels in the sciatic nerve, prefrontal cortex and hippocampus of operated animals. Treatment with fluoxetine or thalidomide reversed crush-induced mechanical hyperalgesia, depressive-like behaviors and the increased TNF-α levels in the sciatic nerve, prefrontal cortex and hippocampus. CONCLUSIONS: The sciatic nerve crush model represents a good model to study to mechanisms underlying both pain and depressive-like behaviors. Furthermore, inhibitors of TNF-α synthesis, like thalidomide, have a potential to treat depressive disorders associated with neuropathic pain.

Fast copper extraction from printed circuit boards using supercritical carbon dioxide.

Technological development and intensive marketing support the growth in demand for electrical and electronic equipment (EEE), for which printed circuit boards (PCBs) are vital components. As these devices become obsolete after short periods, waste PCBs present a problem and require recycling. PCBs are composed of ceramics, polymers, and metals, particularly Cu, which is present in highest percentages. The aim of this study was to develop an innovative method to recover Cu from the PCBs of old mobile phones, obtaining faster reaction kinetics by means of leaching with supercritical CO2 and co-solvents. The PCBs from waste mobile phones were characterized, and evaluation was made of the reaction kinetics during leaching at atmospheric pressure and using supercritical CO2 with H2O2 and H2SO4 as co-solvents. The results showed that the PCBs contained 34.83 wt% of Cu. It was found that the supercritical extraction was 9 times faster, compared to atmospheric pressure extraction. After 20 min of supercritical leaching, approximately 90% of the Cu contained in the PCB was extracted using a 1:20 solid:liquid ratio and 20% of H2O2 and H2SO4 (2.5 M). These results demonstrate the efficiency of the process. Therefore the supercritical CO2 employment in the PCBs recycling is a promising alternative and the CO2 is environmentally acceptable and reusable.

DNA damage in Wistar rats exposed to dithiocarbamate pesticide mancozeb.

Pesticides are used in large amounts in agriculture and the evaluation of their toxic effects is of major concern to public and environmental health. The aim of the present study was to investigate the genotoxic potential of a commercial formulation of the fungicide mancozeb by the micronucleus test in bone marrow and the comet assay in total blood of Wistar rats. Adult male Wistar rats were treated with a solution of mancozeb at a concentration of 40 mg/kg/day, administered intraperitoneally for 18 consecutive days, and compared to a control group. The results indicate that mancozeb induced significantly higher DNA damage as detected by the comet assay and increased the frequency of micronuclei. The results show that mancozeb is genotoxic and may adversely affect the DNA integrity of exposed organisms.

Short-term effect of leflunomide in patients with Takayasu arteritis: an observational study.

OBJECTIVES: To evaluate the efficacy of leflunomide in controlling disease activity in patients with Takayasu arteritis (TA) refractory or intolerant to conventional treatment. METHODS: We conducted a prospective open-label study of 15 TA patients (mean age 36.2 years) with active disease based on clinical assessment, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and magnetic resonance angiography (MRA). Patients received leflunomide 20 mg/day for at least 6 months and were followed up for a mean of 9.1 months. Adverse events attributable to leflunomide were recorded. RESULTS: At baseline, 14 TA patients had active disease despite therapy with corticosteroids and immunosuppressive agents, while one patient had intolerance to current treatment. In the follow-up visit, we found a significant decrease in the frequency of patients with active TA (93% vs. 20%, p = 0.002), in the mean daily dose of prednisone (34.2 vs. 13.9 mg, p < 0.001) and in the median values of ESR (29.0 vs. 27.0 mm/h, p = 0.012) and CRP (10.3 vs. 5.3 mg/L, p = 0.012). Two patients (13.3%) developed new angiographic lesions in the follow-up MRA. Three patients (20%) experienced mild adverse events during the study and none discontinued therapy. CONCLUSIONS: This is the first open-label study to demonstrate improvement in disease activity and acute phase reactants with 20 mg/day of leflunomide in TA patients who were refractory or intolerant to conventional therapy with corticosteroids and immunosuppressive agents. Leflunomide was safe and a steroid-sparing effect was observed. A double-blind controlled study is desirable to confirm this finding.

Manual acupuncture inhibits mechanical hypersensitivity induced by spinal nerve ligation in rats.

Manual acupuncture (MA) has presented analgesic activity against neuropathic pain in patients and animal models, yet a series of questions remain: Is MA effectiveness dependent of acupoint selection or combination? Is it equally efficient when treatment starts on the initial (acute) or sub-chronic phase of spinal nerve ligation (SNL)-induced neuropathy? Is MA effect related to the release of endogenous opioids? Does MA produce similar effects to gabapentin? To answer these questions rats submitted to the L5/L6 SNL injury were treated with unilateral MA (ST36 (Zusanli), SP6 (Sanyingjiao) or ST36+SP6 acupoint stimulation); or with gabapentin (30 mg/kg i.p., used as positive control). Both acupoints have been demonstrated to present analgesic activity and are used in clinical practice and basic science research. In addition, we investigated the influence of naloxone (1 mg/kg i.p., a nonselective opioid receptor antagonist) on MA treatment and also the effect of unilateral ST36+SP6 MA treatment beginning acutely (5 days) or sub-chronically (14 days) after SNL. Our results demonstrate that single or combined unilateral stimulation was able to reduce mechanical hypersensitivity with treatment beginning in both acute and sub-chronic phases of SNL-induced neuropathy; MA effect was blocked by naloxone, and finally; SP6+ST36 MA presented similar effect to gabapentin (30 mg/kg). In conclusion, our results demonstrate, for the first time, that unilateral MA (ST36, SP6 or ST36+SP6) reduces hypersensitivity induced by the SNL with effect dependent of the opioid system and comparable with the one obtained with gabapentin (used as positive control).

New chiral imidazolium ionic liquids from isomannide.

New chiral bis and mono-imidazolium ionic liquids derived from isomannide were synthesized. The structural features of the chiral organic cations impart a special arrangement of the chiral cavity. The new chiral chloride salts of isomannide derivatives are pivotal compounds for the synthesis of different organic ionic liquids. After metathesis different anions were associated to the chiral cations providing a new class of chiral ionic liquids.

Characterization of the volatile fraction emitted by Pinus spp. by one- and two-dimensional chromatographic techniques with mass spectrometric detection.

The chemical composition of the needles of P. pinea, P. pinaster, P. halepensis, P. nigra, P. brutia, P. patula, P. radiata, P. taeda, P. elliotti, P. kesiya, P. sylvestris and P. eldarica was investigated. Headspace solid-phase microextraction and steam distillation extraction were used to collect the volatile fractions. Samples were analyzed using one-dimensional gas chromatography (1D-GC) and comprehensive two-dimensional gas chromatography (GCxGC) associated with a quadrupole and a time-of-flight mass detectors. Results showed that the analytical capabilities of 1D-GC are partially limited by the separation power of the columns. The higher sensibility and the absence of peak skewing of the time-of-flight mass analyzer, with the use of automated peak finding and deconvolution algorithms, allowed for the detection of trace components with qualitative full spectra and the extraction of true mass spectra from coeluting compounds, promoting their reliable identification and thus significantly improving results obtained by 1D-GC/MS, when using a quadrupole mass analyzer. The use of GC x GC resulted in enhanced separation efficiency and increased signal to noise ratio (sensitivity) of the analytes, maximizing mass spectra quality and improving compound detection and identification. This work shows the use of 1D-GC/ToFMS for the analysis of pine needles volatiles, achieving the detection of 177 compounds, that is more than twice the number previously identified by standard 1D-GC/MS. The analysis by GC x GC for the same sample allowed the detection of 212 compounds. The enantioselective GC x GC analysis performed for all the Pinus spp. under study achieved the detection of 422 different compounds. Cross-over phenomena according to operational conditions are highlighted and discussed.

Comprehensive two-dimensional gas chromatography for fingerprint pattern recognition in olive oils produced by two different techniques in Portuguese olive varieties Galega Vulgar, Cobrançosa e Carrasquenha.

For olive oil production a metal hammer-decanter olive processing line was compared to a traditional metal hammer-press line, a discontinuous method which, if properly used, yields high-quality virgin olive oils. Galega, Carrasquenha and Cobrançosa olives (traditional Portuguese varieties) were studied. The analysis of the aroma compounds was performed after headspace-solid phase micro extraction. The analytical results obtained after comprehensive gas chromatography in tandem with time of flight mass spectrometry (GCxGC/ToFMS) for these three different olive oil varieties, from a single year harvest and processed with two different extraction technologies, were compared using statistical image treatment, by means of ImageJ software, for fingerprint recognitions and compared with principal component analysis when the area data of each chromatographic spot of the contour plots were considered. The differences used to classify the olive oils studied under different groups after principal component analysis were observed independently of the treatment used (peak areas or the sum of the pixels counts). When the individual peak areas were considered, more then 75.7% of the total variance is explained by the first two principal components while in the case where the data were subjected to image treatment 84.0% of the total variance is explained by the first two principal components. In both cases the first and second principal components present eigenvalues higher then 1.0. Fingerprint image monitoring of the aroma compounds of the olive oil allowed a rapid differentiation of the three varieties studied as well as the extraction methods used. The volatile compounds responsible for their characterization were tentatively identified in a bi-dimensional polar/non-polar column set in the GCxGC/Tof-MS apparatus. This methodology allowed the reduction of the number of compounds needed for matrices characterization, preserving the efficiency of the discrimination, when compared with the traditional methods where the identification of all peaks is needed.

An assessment of acute biomarker responses in the demersal catfish Cathorops spixii after the Vicuña oil spill in a harbour estuarine area in Southern Brazil.

The Vicuña oil tanker exploded in Paranaguá Bay (South of Brazil), during methanol unloading operations in front of Paranaguá Harbour, on November 15th, 2004, releasing a large amount of bunker oil and methanol. Two weeks after the accident, the acute effects of the Vicuña Oil Spill (VOS) were evaluated in the demersal catfish Cathorops spixii, comparing a contaminated (at the spill site) and a reference site inside the Bay. Data were compared to previous measurements, taken before the accident, in the same species, from the same sites. The physiological biomarkers were the ones that best reflected acute effects of the spill: plasma osmolality, chloride, calcium, magnesium, and potassium. Morphological (liver and gill histopathology) and genetic (piscine micronucleus and DNA strand breaks) biomarkers revealed that damage was already present in fishes from both reference and contaminated sites inside the Bay. Thus, the reference site is not devoid of contamination, as water circulation tends to spread the contaminants released into other areas of the Bay. Acute field surveys of oil spill effects in harbour areas with a long history of contamination should thus be viewed with caution, and whenever possible previous evaluations should be considered for proper appraisal of biomarker sensitivity, especially in mobile bioindicators such as fish.

Comprehensive two-dimensional gas chromatography for fingerprint pattern recognition in cachaça production.

Cachaça samples were studied by means of comprehensive two-dimensional gas chromatography and time-of-flight mass spectrometry (GCxGC/TOFMS) during the fermentation process and after ageing in different wood materials. The analyses of the aroma compounds were performed after headspace-solid phase microextraction method (HS-SPME) using an 85microm polyacrylate (PA) fibre. Fingerprint monitoring of the distillation process allowed the easy determination of the turning points of the process and high-resolution comparison of cabeça (head), coração (core) and cauda (tail) fractions. The ageing process in different wood materials was well characterised through fingerprint similarity observations; in the absence of a suitable metric for expressing the overall similarity, here we use a visual and retention time comparison to identify co-incident peaks and those that differ between samples. For quality control purposes, a simple observation of the contour plots obtained can thus allow the identification of the type of wood used in the ageing process, and the process of ageing, without further statistical treatment or peak identifications. In this manner, peaks, which discriminated most between the different mixtures studied were readily found, i.e. unique compounds were identified in each stage of the distillation process. Approximate first dimension linear retention indices (LRI) for these identified compounds were calculated in a bi-dimensional polar/non-polar column set in the GCxGC experiment and were used in conjunction with mass spectral library searching for tentative identification. Along the progression of the distillation process, 70 compounds appear to visually discriminate between samples and their retention indices are indicated, presenting good correlation with literature data.

Comprehensive two-dimensional gas chromatography/mass spectrometric analysis of pepper volatiles.

The headspace compositions of 13 pepper and peppercorn samples of different species, colloquially also referred to as pepper, were analyzed, and more than 300 compounds were tentatively characterized by means of comprehensive two-dimensional gas chromatography in tandem with flame ionization detection, quadrupole mass spectrometric detection and time-of-flight mass spectrometric detection (GC x GC-FID, GC x GC/qMS and GC x GC/TOFMS, respectively). The analysis of volatile organic compounds (VOCs) was performed after solid-phase microextraction (SPME) using a 75-microm PDMS/DVB fibre. Fingerprint comparison between the three techniques permitted peaks to be assigned in the GC x GC-FID experiment based on the analogous MS analysis, taking into account retention shifts arising from method variations. When using GC x GC/TOFMS, about five times more peaks were identified than in GC x GC/qMS. Retention indices for all peaks were calculated in the bi-dimensional column set comprising of a 5% phenyl polysilphenylene-siloxane primary column and a polyethylene glycol second column. The spectra obtained by both mass detection techniques (qMS and TOFMS) give very similar results when spectral library searching was performed. The majority of the identified compounds eluted as pure components as a result of high-resolution GC x GC separations, which significantly reduces co-elution, and therefore increases the likelihood that pure spectra can be obtained. The differences between TOFMS and qMS (in fast scanning mode) spectra were generally small. Whilst spectral quality and relative ion ratios across a narrow peak (e.g. w(b) approximately 100-150 ms) do vary more for the fast peaks obtained in GC x GC/qMS operation, than with TOFMS, in general adequate spectral matching with the library can be achieved.

Characterization of the volatile fraction emitted by phloems of four pinus species by solid-phase microextraction and gas chromatography-mass spectrometry.

Pine forests constitute some of the most important renewable resources supplying timber, paper and chemical industries, among other functions. Characterization of the volatiles emitted by different Pinus species has proven to be an important tool to decode the process of host tree selection by herbivore insects, some of which cause serious economic damage to pines. Variations in the relative composition of the bouquet of semiochemicals are responsible for the outcome of different biological processes, such as mate finding, egg-laying site recognition and host selection. The volatiles present in phloem samples of four pine species, P. halepensis, P. sylvestris, P. pinaster and P. pinea, were identified and characterized with the aim of finding possible host-plant attractants for native pests, such as the bark beetle Tomicus piniperda. The volatile compounds emitted by phloem samples of pines were extracted by headspace solid-phase micro extraction, using a 2cm 50/30mm divinylbenzene/carboxen/polydimethylsiloxane table flex solid-phase microextraction fiber and its contents analyzed by high-resolution gas chromatography, using flame ionization and a non polar and chiral column phases. The components of the volatile fraction emitted by the phloem samples were identified by mass spectrometry using time-of-flight and quadrupole mass analyzers. The estimated relative composition was used to perform a discriminant analysis among pine species, by means of cluster and principal component analysis. It can be concluded that it is possible to discriminate pine species based on the monoterpenes emissions of phloem samples.

Determination of Cd and Pb in biological reference materials by electrothermal atomic absorption spectrometry: A comparison of three ultrasonic-based sample treatment procedures.

Three different ultrasonic-based sample treatment approaches, the automated ultrasonic slurry sampling, the ultrasonic assisted acid solid-liquid extraction (ASLE) and the enzymatic probe sonication (EPS) were compared and discussed for the determination of Cd and Pb by ET-AAS in biological reference materials. The sample mass chosen to perform the analysis was 10mg and the liquid volume was 1ml of nitric acid 1M. The best results were obtained with the slurry procedure with which it was possible accurate and precise determination of the Cd and Pb content in four of the five reference materials studied. Optimum performance (total metal extraction) of ASLE assisted by ultrasound for Cd was only achieved in two of the four materials assessed whereas total Pb recovery was only possible in three of the five samples. Total extraction with the enzymatic probe sonication was only obtained for Cd in oyster tissue. Neither ASLE nor EPS were able to extract Cd or Pb from spruce needles. Pb concentration obtained after EPS was found to be highly dependent from sample centrifugation speed and time.

Different multidimensional chromatographic approaches applied to the study of wine malolactic fermentation.

Different multidimensional chromatographic techniques were used to study wine aroma pattern changes during malolactic fermentation (MLF). Ethyl lactate enantiomeric ratios were determined using on-line multidimensional gas chromatography. The values found agree with a spontaneous MLF. Off-line multidimensional HPLC/GC was used to deconvolute and enrich the sample and ease enantioselective chromatography. Chiral compound enantiomeric ratio changes during MLF were monitored. Evaluation of enantiomeric ratio changes during MLF has never been studied. (R,R), (S,S) and meso-butane-2,3-diol and pentane-2,4-diol (reported in wines for the first time) were submitted to untrained sensory panel tests. All stereoisomers revealed different sensory notes; pentane-2,4-diol showed an aromatic impact.

Endotoxins, asthma, and allergic immune responses.

Asthma severity depends to a great extent on the levels of endotoxin present in the microenvironment. Although favouring a Th1 cytokine response that could be beneficial to the asthmatic, lipopolysaccharide (LPS) aggravates bronchopulmonary inflammation by several mechanisms. These include neutrophil and eosinophil recruitment, and release by activated macrophages of pro-inflammatory cytokines and nitric oxide. LPS exerts its biological actions through its interaction with CD14. The genetic locus of CD14 is close to the genomic region controlling levels of IgE. A polymorphism in the CD14 promoter region seems to favour high serum IgE levels. Genetic influences may thus control circulating levels of sCD14 and by this mechanism modulate Th1/Th2 balance and IgE synthesis. LPS exposure, although hazardous to the asthmatic, seems to exert a role in the maturation of the immune system in children towards a Th1-skewed pattern.

Complementary use of hyphenated purge-and-trap gas chromatography techniques and sensory analysis in the aroma profiling of strawberries (Fragaria ananassa).

The aroma compositions of Fragaria ananassa varieties are assessed by purge-and-trap high-resolution gas chromatography. Gas chromatography/mass spectrometry and gas chromatography/Fourier transform infrared spectroscopy allow the identification of 93 components from which 21 are for the first time described as constituents of strawberry aroma. Despite the complexity of the aroma, sensory properties are assigned to 40 components as perceived by three independent testers by application of a sniffing technique to the chromatographic effluent. The data are used for comparative sensory characterization of strawberry varieties by means of principal component analysis.

Severe forms of neurocysticercosis: treatment with albendazole.

Study of 22 patients with the severe form of neurocysticercosis treated with albendazole (ABZ) administered in 6 different schedules ranging from 15 to 30 mg/kg/day for 21 to 60 days. Dextrochloropheniramine and ketoprofen were the adjuvant drugs. Multiple symptoms were observed in 90.9% of patients. Intracranial hypertension was manifested in 90.9%. Hydrocephaly occurred in 86.4%. Evolution was satisfactory in 10 patients, 8 died and 4 had sequelae. Tomographic studies showed the appearance of an isolated IVth ventricle in 9 patients, after ventriculoperitoneal shunt, before ABZ treatment in 3 of them, during in 5 and after treatment in one. Median clinical follow-up duration was 10 months for the patients who died and 3-4 years for survivors. In 3 patients there was an increase in cyst size during the administration of the 15 mg/kg/day ABZ dose, which was not observed in any patient when the 30 mg/kg/day dose was used.

[Polyradiculoneuritis and neurocysticercosis: report of a case]. / Polirradiculoneurite e neurocisticercose: relato de caso.

Report of a patient with the hydrocephalic and meningoencephalitic form of neurocysticercosis who simultaneously developed polyradiculoneuropathy and intracranial hypertension syndrome during the first week of treatment with albendazole. Etiologic agents associated with polyradiculoneuropathy related in the literature are cited. Some comments about the possible physiopathogeny of this entity in the presence of cysticercosis are also done. It is mentioned another case who presented polyradiculoneuropathy as the only manifestation of a probable cysticercosis of the nervous system. In this presented case, including the neurocysticercosis and even a mere coincidence of facts, some factors can have a relationship with the appearance of polyradiculoneuropathy as the surgical stress, the stress due to the severity of the clinical picture and the possible side effect of albendazole.