178 research outputs found
Cloning and gene expression analysis of the phospholipase C in wounded spinach leaves during postharvest storage
No full textMembrane destabilization in plant cells can be caused by mechanical injuries, pathogen attacks, environment
stresses or senescence. Wounds represent vulnerable points that may lead to severe damage and
can compromise organ survival rates. Loss of membrane integrity is often associated with lipid peroxidation
or degradation of phospholipids. This work focuses on the isolation of phospholipase C (PLC) in
spinach leaves and the investigation of its role in membrane destabilization during leaf senescence and
after wounding.
Degenerate primers were used to amplify a 270 bp fragment with RT-PCR and a full length SoPLC
mRNA was isolated using a RACE approach. The mRNA was 2290 bp in length and contained an open
reading frame of 1765 bp. The predicted amino acid sequence showed high similarity with PLC2 and
PLC7 of Arabidopsis. Gene expression analyses showed that SoPLC was down-regulated by wounds and
up-regulated by detached induced senescence.
Membrane integrity was evaluated by lipid peroxidation and HPLC phospholipid analyses. Phospholipase
C (PLC) and D (PLD) enzyme activities were determined in detached leaves incubated in the dark at
4 or 20◦C. Lipid peroxidation and enzymatic activities were mainly affected by senescence and temperature
rather than wounds. Lipid peroxidation did not change at 4 ◦C with TBARS values ranging from 3
to 4 nmol g−1 FW. Leaves incubated at 20 ◦C showed an increase of TBARS from 4 to 12 nmol g−1 FW. PLC
and PLD enzymatic activities in leaves incubated at 20 ◦C significantly increased after seven days with
higher values in wounded leaves (3300 pKatmg−1 prot) compared with control (550 pKatmg−1 prot.)
Extraction and Quantification of Azelaic Acid from Different Wheat Samples (Triticum durum Desf.) and Evaluation of Their Antimicrobial and Antioxidant Activities
No full text: Azelaic Acid (AzA) is a 9-carbon atom dicarboxylic acid, with numerous pharmacological uses in dermatology. Its effectiveness in papulopustular rosacea and acne vulgaris, among other dermatological disorders such as keratinization and hyper-pigmentation, is thought to be related to its anti-inflammatory and antimicrobial properties. It is a by-product of Pityrosporum fungal mycelia metabolism but also it is found in different cereals such as barley, wheat, and rye. Diverse topical formulations of AzA exist in commerce, and it is mainly produced via chemical synthesis. In this study we describe the extraction of AzA from whole grains and whole-grain flour (Triticum durum Desf.) through green methods. Seventeen different extracts were prepared and analyzed for their AzA content by HPLC-MS methods and then screened for their antioxidant activity using spectrophotometric assays (ABTS, DPPH, and Folin-Ciocalteu). Minimum-inhibitory-concentration (MIC) assays against several bacterial and fungal pathogens were performed, to validate their antimicrobial activity. The obtained results indicate that whole grain extracts provide a wider spectrum of activity than the flour matrix; in particular, the Naviglio® extract showed higher AzA content, while the hydroalcoholic ultrasound-assisted extract provided better antimicrobial and antioxidant activity. The data analysis was performed using principal component analysis (PCA), as an unsupervised-pattern-recognition technique, to extract useful analytical and biological information
A follow up serological study on Neospora caninum infection in herds affected by abortion in Italy.
No full textStudio di follow up su un approccio combinato, vaccinazione ed eliminazione degli animali immunotolleranti, per il controllo della Diarrea Virale Bovina (BVD)
No full textIntroduction - The control of bovine viral diarrhea (BVD)
infection is based on three main pillars: implementation of
bio-safety measures, test and cull of persistently infected animals
(PI) and adoption of a vaccine protocol to prevent transplacental
infection (fetal protection).
Aim - The aim of this study is to verify, in field conditions,
the efficacy of a combined approach that provides for detection
and removal of PI animals and vaccination with a vaccine
with the claim of fetal protection to BVDV.
Materials and methods - The study lasted 9 years (November
2008-December 2017), was carried out in three farms located
in the Province of Modena (Italy) and involved about
4,800 animals. Anamnestic data pointed out that in all the
farms a BVD vaccination protocol was in place for at least 8
years (2001-2008) with both inactivated and modified-live
vaccines. At the beginning of the study, all animals over 5
months of age were tested, in pool of 20 animals, for the detection
of BVDV by RT-PCR. Subsequently, in the screening
phase, PI research by RT-PCR was performed on all newborn
calves. Moreover, BVDV detection in bulk milk was performed
as well on a four-monthly basis. Regarding vaccination
protocol, the animals were vaccinated with a modified-live
vaccine (Mucosiffa®), namely heifers every six months and
cows in the post-partum.
Results and discussion - During the study, three PI animals
were identified. These animals were born from non-PI cows
vaccinated with an inactivated vaccine. During the period
when the animals were vaccinated with the modified-live
vaccine, no PI animals were detected. All the tests on bulk
milk were negative for BVD antigen.
Conclusion - The combined approach of vaccination with a
modified-live vaccine, associated with test and cull of PI calves,
allowed the control of BVD infection, as demonstrated
by absence of new PI calves and BVDV circulation assessed
by RT-PCR in bulk milk
Leptospira seroprevalence in colombian dairy herds
No full textLeptospirosis in cattle has important economic effects on the infected farms. Moreover, livestock farming is considered a major occupational risk factor for the transmission of Leptospira infection to humans. A survey was performed to determine the overall and within-herd seroprevalence and mapping of different Leptospira serovars in dairy cattle from farms located in some municipalities of the Colombian department of Boyacá. Nine hundred and fifty-nine animals, from 20 unvaccinated and one vaccinated herd, were included in the study. Anti-Leptospira serum antibodies were detected by the microscopic agglutination test (MAT). Only one herd was seronegative. Overall seroprevalence to at least one serovar of Leptospira was 24.1% for unvaccinated animals and 62.3% for animals from the vaccinated herd. A very high within-herd seroprevalence (>60%) was present in 20% of the unvaccinated herds. The presence in the vaccinated herd of 20/398 animals showing high titers, between 1000 and 4000, to at least one serovar of Leptospira suggest that some animals could have been infected. Moreover, due to the presence of seronegative animals, a failure of vaccination immunity or the presence of unvaccinated animals in the vaccinated herd cannot be excluded. In all farms, domestic animals other than cattle were present. Considering the farming practices occurring on dairy farms in the study area, higher hygienic standards and stricter biosecurity measures are suggested
Risposta anticorpale verso BVDV tipi 1,2 e NS 2-3 in manze trattate con vaccini vivi attenuati ed inattivati
No full textA FT-NIR process analytical technology approach for milk renneting control
No full textThe study proposes a process analytical technology (PAT) approach for the control of milk coagulation through near infrared spectroscopy (NIRS), computing multivariate statistical process control (MSPC) charts, based on principal component analysis (PCA). Reconstituted skimmed milk and commercial pasteurized skimmed milk were mixed at two different ratios (60:40 and 40:60). Each mix ratio was prepared in six replicates and used for coagulation trials, monitored by fundamental rheology, as a reference method, and NIRS by inserting a probe directly in the coagulation vat and collecting spectra at two different acquisition times, i.e., 60 s or 10 s. Furthermore, three failure coagulation trials were performed, deliberately changing temperature or rennet and CaCl2 concentration. The comparison with fundamental rheology results confirmed the effectiveness of NIRS to monitor milk renneting. The reduced spectral acquisition time (10 s) showed data highly correlated (r > 0.99) to those acquired with longer acquisition time. The developed decision trees, based on PC1 scores and T2 MSPC charts, confirmed the suitability of the proposed approach for the prediction of coagulation times and for the detection of possible failures. In conclusion, the work provides a robust but simple PAT approach to assist cheesemakers in monitoring the coagulation step in real-time
Novel Potential Antifungal Compounds With Dual Mechanism Of Action Selectively Acting Against Malassezia spp.
No full textMalassezia spp. infections and azole drug resistance phenomena are of great concern in both human and veterinary medicine. Malassezia spp. cause severe human and animal skin disorders, with a zoonotic potential for M. pachydermatis, which could include it on WHO fungal pathogens priority list. Inorganic SeS2 is used as topical treatment, but its mechanism of action on fungal sterol pathways has not been fully revealed, due to the great variability of lipidome among Malasseziomycetes. In this work we evaluated antifungal activity by microdilution broth assay of novel compounds with acyl/selenoureido moieties and primary/secondary sulfonamide groups with a dual mechanism of action: (i) a selective organic selenium fungal toxicity and (ii) the inhibition of a new antifungal target metalloenzyme, the Carbonic Anhydrases (CAs, EC 4.2.1.1). Minimal Inhibitory Concentration (MIC) values of selenium-containing compounds showed very high activity on M. pachydermatis (0,5-3,33 μg/ml) instead of C. albicans and C. glabrata (3,33-256 μg/ml). Suppression of antifungal activity was noted when selenium was replaced with either chalcogen isosteric elements oxygen and sulfur. Compounds library was also tested on M. furfur and M. globosa showing preferential activity on M. pachydermatis, with only a few candidates more active on M. furfur. Cytotoxicity properties of selected compounds against MDBK and HaCat cells were assessed, which showed safety profile at MIC values, better than SeS2. KI values on Malassezia spp. CAs of compounds bearing primary or secondary sulfonamide moiety was in the low-medium nanomolar range, demonstrating a multitarget selective activity on Malassezia spp., probably depending on lipidome constitution
Cerebral perfusion during intermittent hemodialysis in patients with acute kidney injury and advanced liver cirrhosis.
No full textBackground: Cerebral blood flow (CBF) decreases during intermittent hemodialysis (HD) in patients with acute kidney injury (AKI). Since cerebral hemodynamics may be impaired in liver cirrhosis (LC), this condition could aggravate cerebral hypoperfusion during HD in patients with AKI. We examined CBF during the first HD session in patients admitted for AKI, with or without LC. Methods: CBF was examined by measuring middle cerebral artery mean flow velocity (MCAmfv) with transcranial Doppler at baseline and at the end of the first 4-hour HD session in 11 patients with both AKI and LC (median age 69 years, range 40-87, 7 men). Eleven patients with AKI without LC (median age 77 years, range 69-92, 6 men) served as controls. Results: Median net ultrafiltration volume at the end of the HD session was slightly, albeit not significantly, smaller in the patients with LC than in those without (-0.25 kg [range 0.00 to -1.50] vs. -1.00 kg [range 0.00 to -2.00], p = 0.18). At end of HD session, median MCAmfv had decreased by -5.5 cm/s (range -41.3 to 9.9) in the patients with LC, and by -4.5 cm/s (range -11.0 to -2.5) in those without LC (p = 0.79). At end of HD session, the mean MCAmfv of the 2 groups, adjusted for baseline MCAmfv and net ultrafiltration volume, was 25.7 and 21.1 cm/s in AKI patients with and without LC, respectively (difference between groups: 4.6 cm/sec; 95% confidence interval, -3.8 to +13.0). Conclusion: In patients with AKI, concomitant LC does not confer greater vulnerability to cerebral hypoperfusion during intermittent H
Seroprevalence to antigens responsible of respiratory disease in beef cattle: two years of observations in Italian beef herds
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