Struvite precipitation in wastewater treatment plants anaerobic digestion supernatants using a magnesium oxide by-product.

Struvite precipitation is a well-known technology to recover and upcycle phosphorus from municipal wastewater as a slow-release fertiliser. However, the economic and environmental costs of struvite precipitation are constrained by using technical-grade reagents as a magnesium source. This research evaluates the feasibility of using a low-grade magnesium oxide (LG-MgO) by-product from the calcination of magnesite as a magnesium source to precipitate struvite from anaerobic digestion supernatants in wastewater treatment plants. Three distinct LG-MgOs were used in this research to capture the inherent variability of this by-product. The MgO content of the LG-MgOs varied from 42 % to 56 %, which governed the reactivity of the by-product. Experimental results showed that dosing LG-MgO at P:Mg molar ratio close to stoichiometry (i.e. 1:1 and 1:2) favoured struvite precipitation, whereas higher molar ratios (i.e. 1:4, 1:6 and 1:8) favoured calcium phosphate precipitation due to the higher calcium concentration and pH. At a P:Mg molar ratio of 1:1 and 1:2, the percentage of phosphate precipitated was 53-72 % and 89-97 %, respectively, depending on the LG-MgO reactivity. A final experiment was performed to examine the composition and morphology of the precipitate obtained under the most favourable conditions, which showed that (i) struvite was the mineral phase with the highest peaks intensity and (ii) struvite was present in two different shapes: hopper and polyhedral. Overall, this research has demonstrated that LG-MgO is an efficient source of magnesium for struvite precipitation, which fits the circular economy principles by valorising an industrial by-product, reducing the pressure on natural resources, and developing a more sustainable technology for phosphorus recovery.

Preparation and physicochemical characterization of deep eutectic solvents and ionic liquids for the potential absorption and biodegradation of styrene vapors.

Styrene emissions can be treated by physicochemical, biological, or physicochemical/biological means. Due to its low solubility in water an alternative to eliminate styrene emissions from air is the use of two-phase partitioning bioreactors (TPPBs) which comprised a hydrophobic non-aqueous phase (NAP) which can improve mass transfer of styrene. This study was devoted to prepare and evaluate the main physicochemical characteristics of novel NAPs such as Ionic liquids (ILs), Deep Eutectic Solvents (DESs) and Natural Deep Eutectic Solvents (NADEs) as well as their toxicity and biodegradability to treat styrene vapors. Absorption experiments of styrene showed that the best NAPs were the DESs formed with Tetrabutylammonium bromide and decanoic acid and the ILs [C6mim][FAP], [C4mim] [NTf2] and [C4mim] [PF6], since they presented a styrene partition coefficient between 0.0015 and 0.0041. Finally, the IL [C6mim][FAP] was used as a NAP in a TPPB batch process given its high styrene affinity, low solubility in water and non-biodegradability; styrene mineralization was three times higher in the TPPB compared with the control. ILs are potential adjuvant phases in biological degradation systems, as well as other solvents like DESs and NADESs.

Efficacy of transcutaneous perineal electrostimulation versus intracavitary anal electrostimulation in the treatment of urinary incontinence after a radical prostatectomy: randomized controlled trial study protocol.

BACKGROUND: Radical prostatectomy is the gold standard treatment for men with localized prostate cancer. This technique is associated with post-operative urinary incontinence. Pelvic floor physiotherapy is a conservative, painless and economical treatment for this specific situation. Kegel exercises and perineal electrostimulation are common techniques to train pelvic floor muscles. The perineal electrostimulation can be applied to the patient with surface electrodes or by an intra-cavitary anal probe. This study proposes that transcutaneous perineal electrostimulation is as effective as intra-cavitary electrostimulation in reducing urinary incontinence secondary to radical prostatectomy. The main objective is to compare the efficacy of the treatment with transcutaneous perineal electrostimulation versus the same intra-cavitary treatment to reduce the magnitude of urinary incontinence after radical prostatectomy, and the impact on the quality of life. METHODS: This single-blind equivalence randomized controlled trial will include 70 man who suffer urinary incontinence post radical prostatectomy. Participants will be randomized into surface electrodes group and intra-anal probe group. The groups will receive treatment for 10 consecutive weeks. Outcomes include changes in the 24-h Pad Test, and ICIQ-SF, SF-12 and I-QoL questionnaires. Clinical data will be collected at baseline, 6 and 10 weeks after the first session, and 6 months after the end of treatment. DISCUSSION: The results will allow us to prescribe the most beneficial perineal electrostimulation technique in the treatment of urinary incontinence derived from radical prostatectomy. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03587402. 27/06/2018.

Urinary sepsis after endourological ureterorenoscopy for the treatment of lithiasis. / Sepsis urinaria tras tratamiento endourológico de la litiasis por ureterorrenoscopia.

OBJECTIVE: To evaluate the incidence, clinical presentation and factors associated with the development of urinary sepsis after performing ureterorenoscopy. MATERIAL AND METHODS: Retrospective study of patients undergoing ureterorenoscopy for the treatment of lithiasis between July 2015 and October 2017. Patients who developed urinary sepsis during the 30 days following the intervention were identified. Personal, clinical, surgical and microbiological backgrounds were collected. Statistical analysis was performed with the Chi squared test (or Fisher's exact test), Student's t (or U Mann-Whitney) or logistic regression as appropriate. RESULTS: 246 ureterorenoscopies were performed, 184 (74.8%) on ureteral stones and 62 (25.2%) on kidney stones, with a mean age of 52 (44.5-59.5) years. After procedure, 18 (7.3%) patients developed urinary sepsis, 10 of them (55.5%) occurred in the first 24h. The urine culture showed enterobacteria (61.1%) and enterococci (38.9%). The antibiogram showed greater sensitivity to nitrofurantoins (100%) and quinolones (72%). The statistical analysis showed that female sex, the clinical debut of urolithiasis as urinary sepsis, having received antibiotic or having required urinary diversion by a double J during debut, positive presurgical uroculture and the persistence of residual lithiasis after surgery were significantly associated (P<.05) with the development of urinary sepsis after ureterorenoscopy. CONCLUSION: Urinary sepsis is a complication that appears after performing ureterorenoscopy, especially in female patients with a history of urinary sepsis, antibiotic therapy, double J, previous positive urine culture or residual lithiasis after the procedure.

Cytokine distribution in mothers and breastfed children after omega-3 LCPUFAs supplementation during the last trimester of pregnancy and the lactation period: A randomized, controlled trial.

OBJECTIVE: To determine whether maternal diet supplementation with omega-3 long chain polyunsaturated fatty acids (omega-3 LC-PUFAs) during the last trimester of pregnancy and the breastfeeding period influences the levels of inflammatory cytokines in mother and infants. MATERIAL AND METHOD: This registered, double-blind randomized study included 46 pregnant women, who were randomly allocated to either an experimental group receiving 400mL/day of a fish oil-enriched dairy drink [320mg docosahexaenoic acid (DHA) + 72mg eicoapentaenoic acid] (FO group, n = 24) or to a control group receiving 400mL/day of a non-supplemented dairy drink (CT group, n = 22), from week 28 of pregnancy until the fourth month of lactation. During the study, maternal dietary patterns were monitored by a nutritionist, who encouraged compliance with current recommendations of fatty acids intake. DHA concentrations and cytokine levels (GM-CSF, IL-2, IL-4, IL-6, IL-10, INF-γ and TNF-α) were measured in maternal plasma at the moment of recruitment and in maternal (n = 46) and infant (n = 46) plasma at birth and 2.5 months after birth. RESULTS: Maternal plasmatic IL-4 levels were higher in FO than in CT subjects (p = 0.009). Additionally, a tendency was observed to higher IL-10 and IL-2 in the FO group. Plasmatic IL-6 however, was higher in CT mothers (p = 0.001). TNF-α was higher in CT infants at birth and 2.5 months after birth (p = 0.005). An analysis of possible relationships between DHA and the concentrations of different cytokines revealed negative correlation between maternal plasmatic IL-6 and DHA (higher plasmatic DHA corresponded to lower IL-6). CONCLUSIONS: Maternal dietary omega-3 LC-PUFAs supplementation during critical periods like pregnancy, lactation and early newborn development may influence the levels of certain inflammatory cytokines, reducing pro-inflammatory cytokines and promoting an anti-inflammatory "environment".

Purification process for MUFA- and PUFA-based monoacylglycerols from edible oils.

Important health benefits have been attributed to monoacylglycerols (MAGs) due to their various physiological functions, owing to which they become candidates for use as functional foods in order to prevent the onset of certain diseases such as colon cancer. In this work, six edible oils, namely: olive, linseed, sunflower, evening primrose, DHASCO® and ARASCO® have been processed to obtain different MUFA- and PUFA- based MAGs. First, the oils were hydrolyzed by means of an enzymatic process using porcine pancreatic lipase and then the reaction products were fractionated by using a liquid chromatography column containing silica gel as stationary phase in order to purify the MAGs-enriched fraction. A second chromatography process was performed using silver nitrate coated silica gel as stationary phase, in order to obtain the different MUFA- and PUFA-based MAGs from the corresponding oils. Overall, MAGs based on oleic, linoleic, α-linolenic, γ-linolenic, arachidonic and docosahexaenoic acids have been isolated in high yields and purities (92.6, 97.4, 95.3, 90.9, 100 and 95.3% purity, respectively). Positional distribution was determined by means of 1H NMR, which revealed a mix of 1(3) and 2-MAGs in variable proportions in the different MAGs.

Oregano Essential Oil as an Antimicrobial and Antioxidant Additive in Food Products.

Food consumers and industries urged the need of natural alternatives to assure food safety and quality. As a response, the use of natural compounds from herbs and spices is an alternative to synthetic additives associated with toxic problems. This review discusses the antimicrobial and antioxidant activity of oregano essential oil (OEO) and its potential as a food additive. Oregano is a plant that has been used as a food seasoning since ancient times. The common name of oregano is given to several species: Origanum (family: Lamiaceae) and Lippia (family: Verbenaceae), amongst others. The main compounds identified in the different OEOs are carvacrol and thymol, which are responsible for the characteristic odor, antimicrobial, and antioxidant activity; however, their content may vary according to the species, harvesting season, and geographical sources. These substances as antibacterial agents make the cell membrane permeable due to its impregnation in the hydrophobic domains, this effect is higher against gram positive bacteria. In addition, the OEO has antioxidant properties effective in retarding the process of lipid peroxidation in fatty foods, and scavenging free radicals. In this perspective, the present review analyzes and discusses the state of the art about the actual and potential uses of OEO as an antimicrobial and antioxidant food additives.

Characterization of goat colostrum oligosaccharides by nano-liquid chromatography on chip quadrupole time-of-flight mass spectrometry and hydrophilic interaction liquid chromatography-quadrupole mass spectrometry.

A detailed qualitative and quantitative characterization of goat colostrum oligosaccharides (GCO) has been carried out for the first time. Defatted and deproteinized colostrum samples, previously treated by size exclusion chromatography (SEC) to remove lactose, were analyzed by nanoflow liquid chromatography-quadrupole-time of flight mass spectrometry (Nano-LC-Chip-Q-TOF MS). Up to 78 oligosaccharides containing hexose, hexosamine, fucose, N-acetylneuraminic acid or N-glycolylneuraminic acid monomeric units were identified in the samples, some of them detected for the first time in goat colostra. As a second step, a hydrophilic interaction liquid chromatography coupled to mass spectrometry (HILIC-MS) methodology was developed for the separation and quantitation of the main GCO, both acidic and neutral carbohydrates. Among other experimental chromatographic conditions, mobile phase additives and column temperature were evaluated in terms of retention time, resolution, peak width and symmetry of target carbohydrates. Narrow peaks (wh: 0.2-0.6min) and good symmetry (As: 0.8-1.4) were obtained for GCO using an acetonitrile:water gradient with 0.1% ammonium hydroxide at 40°C. These conditions were selected to quantify the main oligosaccharides in goat colostrum samples. Values ranging from 140 to 315mgL(-1) for neutral oligosaccharides and from 83 to 251mgL(-1) for acidic oligosaccharides were found. The combination of both techniques resulted to be useful to achieve a comprehensive characterization of GCO.

Factores de riesgo para alteraciones de la densidad mineral ósea en mujeres posmenopáusicas / Risk factors for alterations in bone mineral density in postmenopausal women

OBJETIVO: Determinar los factores de riesgo para presentar alteraciones de la densidad mineral ósea (DMO) en mujeres posmenopáusicas atendidas en la consulta de Menopausia y Climaterio de la Maternidad "Dr. Armando Castillo Plaza", de Maracaibo, Venezuela. MÉTODOS: Investigación descriptiva, con diseño no experimental y transeccional, donde se evaluó la DMO y los factores de riesgo para presentar osteoporosis en 60 mujeres posmenopáusicas. RESULTADOS: La medición de la DMO resultó en promedio de 957,45 ± 149,95 y 905,00 ± 151,25 gramos, con índices T de -0,52 ± 1,66 y -0,55 ± 2,67 en columna vertebral y cuello femoral, respectivamente. Se estableció una prevalencia de osteoporosis del 10% y 5%, y de osteopenia del 43,3% y 50% en columna vertebral y fémur, respectivamente. La menopausia quirúrgica (OR [95% CI] = 4,75 [1,58-14,25]; p=0,004), el consumo excesivo de café (OR [95% CI] = 3,20 [1,40-7,10 1]; p=0,000) o gaseosas (OR [95% CI] = 2,50 [1,18-5,60]; p=0,002), tabaquismo (OR [95% CI] = 1,70 [1,102,80]; p=0,013) y la ausencia de suplementación de calcio más vitamina D (OR [95% CI] = 1,70 [1,00-2,80]; p=0,019) resultaron ser factores significativamente asociados al diagnóstico de osteoporosis u osteopenia (p<0,05). CONCLUSIÓN: Las mujeres posmenopáusicas evaluadas presentan una alta prevalencia de alteraciones en la DMO, principalmente osteopenia, y factores de riesgo para presentar Osteoporosis.

AIM: To determine risk factors for presenting alterations in bone mineral density (BMD) in postmenopausal women attending the consultation of Menopause and Menopause Maternity "Dr. Armando Castillo Plaza", Maracaibo, Venezuela. METHODS: A descriptive research with non-experimental and transactional design where was evaluated BMD and risk factors for developing osteoporosis in 60 postmenopausal women. RESULTS: The BMD measurement was averaged in 957.45 ± 149.95 and 905.00 ± 151.25 grams; with T indexes of -0.52 ± 1.66 and -0.55 ± 2.67 in spine and femoral neck, respectively. The prevalence of osteoporosis was 10% and 5%, while osteopenia was 43.3% and 50% in spine and femur, respectively. Surgical menopause (OR [95% CI] = 4.75 [1.58 to 14.25]; p=0.004); excessive coffee consumption (OR [95% CI] = 3.20 [1,40- 7.10 1]; p=0.000) or gaseous beverages (OR [95% CI] = 2.50 [1.18 to 5.60]; p=0.002); smoking (OR [95% CI] = 1.70 [1.10 to 2.80]; p=0.013) and the absence of supplemental calcium plus vitamin D (OR [95% CI] = 1.70 [1.00 to 2.80]; p=0.019) were be factors significantly associated with the diagnosis of osteoporosis or osteopenia (p<0.05). CONCLUSION: The evaluated postmenopausal women have a high prevalence of abnormal BMD, especially osteopenia, and risk factors for developing osteoporosis.

Colostrum composition of Santa Inês sheep and passive transfer of immunity to lambs.

This study aimed to analyze the chemical composition and the IgG concentration of the colostrum, transitional milk, and mature milk of Santa Inês ewes as well as the transfer of passive immunity to lambs. Thirty-two pregnant ewes and 38 lambs were used. Ewes were milked immediately after lambing and at 12, 24, 36 h and 10 d postpartum. Colostrum was provided to the lambs at 40±15 min (mean±SE) after birth and then at 30-min intervals for obtaining the intake closest to 10% of body weight, and transitional milk was provided ad libitum. Blood from the lambs was collected 36 h after birth for measuring the serum concentrations of IgG, total protein, albumin, and gamma-globulin. The production was lower in primiparous than in multiparous ewes with body condition score (BCS)<2.75, but did not differ between primiparous and multiparous with BCS≥2.75 (interaction parity and BCS). The IgG concentration and fat, protein, lactose, and defatted dry extract percentages were not affected by the BCS of the ewe at lambing or by the parity. The total solids percentage in the colostrum was higher in ewes with BCS<2.75 (interaction BCS and time). The production and the protein, total solid, and defatted dry extract percentages showed quadratic behavior, the fat percentage decreased linearly, and the lactose percentage increased linearly with time postpartum. The IgG concentration in the colostrum was not correlated with the ewe's weight or BCS at the time of lambing. Moreover, the parity, the BCS, the ewe's type of gestation, and the lamb's sex did not influence the serum concentrations of IgG, total protein, albumin, and gamma-globulin in lambs. Adequate passive immune transfer (PIT) was observed in lambs for which the IgG intake was higher than 30 g. Failure in PIT was observed in 39.5% of lambs when considering a serum IgG concentration lower than 15 mg/mL and in 21% when considering a serum total protein concentration lower than 45 mg/mL. The mean apparent efficiency of absorption was 38.10%, with values between 0.02% and 98.80%. The serum IgG concentration was correlated with the total protein concentration (according to the enzymatic colorimetric method), the gamma-globulin concentration, and the absorption efficiency. The extreme variation on apparent efficiency of absorption may have an effect on the success of PIT. Lambs should consume at least 30 g of IgG in the first 24 h of life to ensure adequate PIT.

A HPLC-SRM-MS based method for the detection and quantification of methotrexate in urine at doses used in clinical practice for patients with rheumatological disease: a potential measure of adherence.

Rheumatoid arthritis (RA) is a common autoimmune disease that causes significant disability and reduced life expectancy. The folate antagonist methotrexate (MTX) is first-line therapy for RA when used weekly at low doses (5-25 mg). However, the true rate of adherence to MTX is uncertain. This is in part due to the different methods of measurement of adherence employed with no biochemical test currently available to determine adherence to low dose MTX. Common methods of MTX measurement include immunoassays in patients with high dose therapy, but these assays cross-react with MTX metabolites and lack the sensitivity required to measure adherence to low dose MTX. HPLC-SRM-MS (selected reaction monitoring-mass spectrometry) has several theoretical advantages over immunoassays with improved specificity, minimal cross-reaction and higher sensitivity. The aim of this study was to develop an assay to measure MTX and its major metabolite 7-OH-MTX in urine as a tool to monitor adherence to low dose MTX in clinic. As a proof of concept, urine samples from 4 participants with RA were measured after directly observed therapy. The assay showed improved sensitivity compared to that reported by immunoassays, with low carryover and high within-run precision. In participant samples, MTX was measurable in the urine for up to 105 hours after administration and 7-OH-MTX was detectable up to 98 hours after administration, suggesting that this assay is suitable for the measurement of adherence to therapy. The assay requires minimal sample preparation and can be adopted by other laboratories with minimal study set up.

Nab-paclitaxel: a flattering facelift.

The application of nanotechnology in oncology has increased the efficacy and efficiency of some cytotoxic agents. The paradigm in this field is nab-paclitaxel, a soluble form of paclitaxel that is linked to albumin nanoparticles. The development of nanotechnology as a delivery system for paclitaxel has provided better pharmacokinetic and pharmacodynamic characteristics, neutralizing its hydrophobicity. This procedure significantly improves the treatment of metastatic breast cancer compared to conventional paclitaxel-based therapies, including other type of cancers such as metastatic pancreatic cancer, stage IIIB-IV non-small cell lung cancer (NSCLC) and metastatic melanoma. In these last cases, significant differences were found in primary end-points for patients treated with nab-paclitaxel-based chemotherapy compared to those treated with conventional treatments. The application of nanotechnology in cancer treatment may also improve the efficacy of other known drugs, as a result of improved pharmacokinetic and pharmacodynamic profiles, similarly to paclitaxel.

Evaluation of THP-1 cell line as an in vitro model for long-term safety assessment of new molecules.

OBJECTIVE: Monitoring the chronic and subacute toxicity is essential in the development of new cosmetic ingredients. In response to the present lack of validated alternative methods, we developed an in vitro model for repeated dose cytotoxicity on THP-1 cells. METHODS: Cultured in suspension, cells were treated with chemicals for 14 days with a frequency of three applications per week, and cell viability was determined by MTT assay. We first investigated the long-term effects of chemicals that induce different kinds of cytotoxicity: Paraquat (PQ), 3-Nitropropanoic acid (3-NPA) and sodium dodecyl sulphate (SDS). From acute studies, doses between 1 and 10 µg ml(-1) were chosen to perform our subacute cytotoxicity assay. Comparative genotoxicity evaluations were made with H2 O2 or Paraquat treated TPH-1 cells. Comet assays were performed at 1 h (4°C); after a 24-h recovery period (37°C); and finally, after a long-term period of treatment (14 days, 37°C).Once adapted to plant extracts or highly diluted molecules, some of our cosmetic compounds were tested with this model. RESULTS: As expected, after 14 days of treatment with Paraquat, cell viability rates dramatically decreased for doses as low as 3 µg ml(-1) , whereas 10 µg ml(-1) of 3-NPA and SDS did not induce more than 44% of cell death. Surprisingly, after subacute treatment, comet assay results revealed a dose-dependent increase in tail moments for Paraquat, whereas those of H2 O2 remained low. Moreover, all our compounds tested at 0.5-5 µg ml(-1) were classified as safe, even with a cut-off at 90% of cell viability. CONCLUSION: In conclusion, this assay could be of interest for subacute cytotoxicity and genotoxic assessment of daily and topically applied products and suggests that PQ is a choice worthy positive control.

Interactions of sulforaphane and dimethyl sulfoxide with methyl methanesulfonate, urethane, 4-nitroquinoline-1-oxide and hydrogen peroxide in the Drosophila melanogaster wing spot test.

Sulforaphane (SF) is an isothiocyanate present in Brassicaceae, vegetables that induce the detoxification of electrophiles and reactive oxygen species. SF has been correlated with chemoprevention mechanisms against degenerative diseases. We tested if the SF had an effect against methyl methanesulfonate (MMS), urethane (URE), 4-NQO and H(2)O(2). SF (>95% purity, 0.14, 0.28, 0.56 mM) was diluted in a DMSO/Tw80/EtOH mixture (DTE) corresponding to 25, 50, 100% of lyophilized broccoli. The SF treatment (0.14 mM) was positive for small spots in the ST cross and negative in the HB cross. In the HB cross, SF (0.28 mM) was genotoxic. In the ST cross, the SF treatments showed a tendency to reduce the genotoxic damage caused by MMS, which could be explained by the radical scavenging action of the DTE mixture. In the ST cross, the frequency of small spots in the SF 0.14 mM/URE treatment was similar to that of Water/URE, which can be explained by a DTE and SF scavenger action. In both crosses, the results for the direct oxidants, 4-NQO and H(2)O(2), were different and must be related to differential modulation of CYPs expression and the SF and DTE scavenger properties.

Impact of defoliation intensities on plant biomass, nutrient uptake and arbuscular mycorrhizal symbiosis in Lotus tenuis growing in a saline-sodic soil.

The impact of different defoliation intensities on the ability of Lotus tenuis plants to regrowth, mobilise nutrients and to associate with native AM fungi and Rhizobium in a saline-sodic soil was investigated. After 70 days, plants were subjected to 0, 25, 50, 75 and 100% defoliation and shoot regrowth was assessed at the end of subsequent 35 days. Compared to non-defoliated plants, low or moderate defoliation up to 75% did not affect shoot regrowth. However, 100% treatment affected shoot regrowth and the clipped plants were not able to compensate the growth attained by non-defoliated plants. Root growth was more affected by defoliation than shoot growth. P and N concentrations in shoots and roots increased with increasing defoliation while Na(+) concentration in shoots of non-defoliated and moderately defoliated plants was similar. Non-defoliated and moderately defoliated plants prevented increases of Na(+) concentration in shoots through both reducing Na(+) uptake and Na(+) transport to shoots by accumulating Na(+) in roots. At high defoliation, the salinity tolerance mechanism is altered and Na(+) concentration in shoots was higher than in roots. Reduction in the photosynthetic capacity induced by defoliation neither changed the root length colonised by AM fungi nor arbuscular colonisation but decreased the vesicular colonisation. Spore density did not change, but hyphal density and Rhizobium nodules increased with defoliation. The strategy of the AM symbiont consists in investing most of the C resources to preferentially retain arbuscular colonisation as well as inoculum density in the soil.

Combinatorial gene therapy induces regression of hepatic encephalopathy.

Capillarization of the sinusoid impedes the clearance of neurotoxic substances in liver fibrosis. These events may result in hepatic encephalopathy. Neurological and hepatic features of rats after bile duct ligation (BDL) supplemented with Manganese (BDL+Mn(2+)) were examined. The 4-week-old BDL rats had elevated levels of ammonia and were concomitantly fed with 1 mg ml(-1) of MnCl(2) in drinking water (BDL/Mn(+2)). Five out of fifteen rats were killed and the serum, liver and brain tissue (striatum and substantia nigra) were recovered. Of the remaining BDL/Mn(+2)-cirrhotic animals (n=10), five were injected with a combination of Adenovirus-human plasminogen activator (Ad-huPA) and Adenovirus-matrix metalloproteinase-8 (Ad-MMP-8) (3 × 10(11)+1.5 × 10(11) vector particles per kg), and five with 4.5 × 10(11) vector particles per kg of Adenovirus-ß-galactosidase (Ad-ß-Gal). This treatment was carried on for 10 days. The BDL/Mn(+2) rats displayed tremor, rigidity and gait abnormalities, which improved notably with combinatorial gene therapy, as well as motor coordination. Liver fibrosis was evidently less after treatment with Ad-huPA+Ad-MMP-8 (25%). In the brain (striatum), Ad-huPA+Ad-MMP-8 treatment rendered higher concentrations of dopamine compared with Ad-ß-Gal-treated encephalopathic rats (210 and 162 ng g(-1) of tissue, respectively). The BDL/Mn(+2) animals and controls treated with Ad-ß-Gal showed abnormal morphology in astrocytes (gliosis) in striatum and substantia nigra, in which expressions of green fibrillar acidic protein and tyrosine hydroxylase were altered. These abnormalities decreased with Ad-huPA+Ad-MMP-8 treatment. Importantly, the latter animals showed an increment in sprouting of nervous fibers in substantia nigra. Combinatorial gene therapy improves neuroanatomical and neurochemical characteristics similar to human hepatic encephalopathy.

In vitro inhibition of lipopolysaccharide and mycobacterium bovis bacillus Calmette Guérin-induced inflammatory cytokines and in vivo protection from D-galactosamine/LPS -mediated liver injury by the medicinal plant Sclerocarya birrea.

Sclerocarya birrea is a medicinal plant used for the treatment of inflammatory- and bacterial-related diseases. The present study investigated in vitro and in vivo the effects of the stem bark methanol extract of S. birrea. Nitrite, TNF, IL-1beta, IL-6 and IL-12p40 production by bone marrow-derived macrophages (BMDM) pre-incubated with or without S. birrea, and stimulated with Lipopolysaccharide (LPS) or infected with live Mycobacterium bovis Bacillus Calmette Guérin (BCG) was evaluated. S. birrea extract inhibited, in a concentration-dependent manner, nitrite, TNF, IL-1beta, IL-6 and IL-12p40 production by BMDM stimulated with LPS or infected with live BCG. The iNOS expression was reduced by S. birrea after stimulation of BMDM with LPS. In addition, S. birrea inhibited the nuclear factor kB (NF-kB) activation by both LPS and BCG. The effects of the plant extract were also evaluated in an in vivo model of liver injury induced by D-galactosamine/LPS (D-GalN/LPS) administration in mice. S. birrea limited D-GalN/LPS-liver injury as assessed by a reduction in transaminases and TNF, IL-1beta, IL-6 serum levels, and translocation of NF-kB to the nucleus. Taken together, our data indicate that stem bark methanol extract of S. birrea possesses anti-inflammatory properties by inhibiting NF-kB activation and cytokine release induced by inflammatory or infectious stimuli.

Activity of ciprofloxacin and levofloxacin in experimental pneumonia caused by Klebsiella pneumoniae deficient in porins, expressing active efflux and producing QnrA1.

The objective of this study was to evaluate the activities of ciprofloxacin and levofloxacin in a murine model of pneumonia caused by Klebsiella pneumoniae C2 (with altered GyrA, deficient in porins and expressing active efflux of quinolones) and the transconjugant C2pMG252 derived from it and expressing the qnrA1 determinant. MICs and MBCs of the two quinolones were determined according to CLSI guidelines. Time-kill curves (at 1x and 4x MIC) were also performed to assess bactericidal activity. An experimental model of pneumonia in mice was evaluated. Groups of 15 mice were infected with either strain and treated with ciprofloxacin (80 mg/kg/day) or levofloxacin (100 mg/kg/day). Control non-treated animals were also evaluated. In the case of strain C2, log(10) CFU/g of lung in non-treated animals was 9.16 +/- 2.16. This value was reduced to 3.53 +/- 1.04 (p <0.001) and 3.38 +/- 0.46 (p <0.001) in animals treated with ciprofloxacin or levofloxacin, respectively. Percentages of surviving mice were 26.7% (control group) and 100% (both ciprofloxacin and levofloxacin; p <0.001 vs. controls). Bacterial counts (log(10) CFU/g) in lungs of animals infected with strain C2pMG252 were 9.65 +/- 2.49 in non-treated animals and 7.74 +/- 2.67 and 7.57 +/- 3.84 for those treated with ciprofloxacin or levofloxacin, respectively (p >0.05 vs. control group). Of non-treated animals infected with strain C2pMG252, 14.3% survived. Ciprofloxacin and levofloxacin improved the survival in these mice (53.3% for both antimicrobials, p 0.03). In conclusion, the expression of qnrA1 in K. pneumoniae with additional mechanisms of resistance causes decreased efficacy of fluoroquinolones in a pneumonia model in mice.

Fermentation characteristics and microbial growth promoted by diets including two-phase olive cake in continuous fermenters.

Two-phase olive cake (2POC) is the by-product obtained from the so called 'two-phase' procedure to extract olive oil by mechanical methods. After the mechanical extraction the 2POC is dried and most of the remaining oil extracted by chemical means. The production of the crude by-product may reach more than 4 millions t/year in Spain (MAPA, 2003), most of it in areas (Southeast) with shortage of pastures and conventional feeds for ruminants. Six continuous fermenters, inoculated with ruminal liquor from wethers or goats, were fed lucerne hay (LH); LH plus a concentrate including dried two-phase olive cake (LHCO) and; diet LHCO added with polyethylene glycol (LHCOP). The highest pH values and ammonia N (NH(3)-N) output were found in fermenters fed diet LH (6.19 and 6.35 for pH, and 53.7 and 68.9 mg NH(3)N/day, respectively, in fermentes inoculated with rumen liquor from sheep and goats) without differences due to the inoculum origin. The digestibility of carbohydrates (CHO) was affected (p < 0.001) by inoculum (67.0 and 58.8%, respectively, for goats and wethers) the lowest values being for diet LHCO (53.2 and 57.0% with inoculum from wethers and goats, respectively). The main volatile fatty acid (VFA) was acetic acid with higher (p < 0.01) values in fermenters with inoculum from goats than from wethers (80.2 and 63.0 mmol/day respectively). The efficiency of bacterial protein synthesis (EBS) was not different (p > 0.05) with inoculum from wethers and goats [26.4 and 28.1 and 35.2 and 33.5 g bacterial N/kg digested CHO, respectively, obtained by using diamino pimelic acid (DAPA) and purine bases (PB) as microbial markers]. The lowest (p < 0.05) values were found in fermenters fed diets LHCOP and LH, estimated, respectively, from DAPA and PB (21.9 and 29.0 g bacterial N/kg digested CHO). The substitution of a part of lucerne hay by a concentrate including dried 2POC does not seem to cause important differences in efficiency of VFA production. Results concerning bacterial protein synthesis are not so clear since values estimated from DAPA and PB did not show similar trends. Neither inoculum origin nor PEG had important effects on fermentation characteristics. The DAPA could be an adequate marker in continuous fermenters, with comparative purposes, as protozoa are not present and, estimated protein synthesis values are similar to those obtained in vivo with similar diets using urinary excretion of PD (Yáñez Ruiz et al., 2004b). Further research is needed to state the optimal proportion of 2POC in practical diets for ruminants at both maintenance and production stages.