Evaluation of the protection elicited by direct and indirect exposure to live attenuated infectious laryngotracheitis virus vaccines against a recent challenge strain from the United States

In a recent study (Oldoni & García, 2007), some field strains of infectious laryngotracheitis viruses (ILTV) were characterized as genotypically different (group VI) from ILT vaccine strains. The objective of this study was to evaluate the protection elicited by one chicken embryo origin (CEO) and one tissue culture origin (TCO) vaccine against a field isolate from group VI after direct and indirect exposure to ILTV live attenuated vaccines. In phase 1 of the experiment, non-vaccinated chickens were placed into contact with the eye drop vaccinates for a period of four weeks after vaccination. Transmission of the vaccine virus to these in-contact birds was demonstrated by real time PCR and antibody production, although the in-contact birds did not become protected against disease when subsequently challenged in phase 2 of the experiment. This emphasized the importance of uniform vaccination to obtain adequate protection, both to avoid the occurrence of susceptible chickens, and to minimize the potential for reversion to virulence of live-attenuated vaccines. In phase 2, protection against challenge with a group VI field virus was assessed four weeks after vaccination by scoring clinical signs and mortality, and quantifying weight gain. Sentinel birds were added to the groups one day after challenge to assess shedding of challenge virus, using real time PCR and virus isolation, during the period 2 to 12 days post challenge. The results showed that the CEO and TCO eye drop-vaccinated chickens were protected against challenge with the group VI virus, even though it was genetically different from the vaccine strains, and that challenge virus was not transmitted from these protected birds to the sentinels.     

Antioxidant activity by DPPH assay of potential solutions to be applied on bleached teeth

The aim of this study was to assess, using the DPPH assay, the antioxidant activity of several substances that could be proposed to immediately revert the problems caused by bleaching procedures. The percentage of antioxidant activity (AA%) of 10% ascorbic acid solution (AAcidS), 10% ascorbic acid gel (AAcidG), 10% sodium ascorbate solution (SodAsS), 10% sodium ascorbate gel (SodAsG), 10% sodium bicarbonate (Bicarb), Neutralize(?) (NE), Desensibilize(?) (DES), catalase C-40 at 10 mg/mL (CAT), 10% alcohol solution of alpha-tocopherol (VitE), Listerine(?) (LIS), 0.12% chlorhexidine (CHX), Croton Lechleri (CL), 10 % aqueous solution of Uncaria Tomentosa (UT), artificial saliva (ArtS) and 0.05% sodium fluoride (NaF) was assessed in triplicate by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free radical assay. All substances exhibited antioxidant activity, except for CL. AAcidS, AAcidG and VitE exhibited the highest AA% (p<0.05). On the contrary, CHX, NE, LIS and NaF showed the lowest AA% (p<0.05). In conclusion, AAcidS, AAcidG, SodAsS, SodAsG and VitE presented the highest antioxidant activity among substances tested in this study. The DPPH assay provides an easy and rapid way to evaluate potential antioxidants.     

Development and validation of a real-time Taqman PCR assay for the detection and quantitation of infectious laryngotracheitis virus in poultry

In this study, the development and validation of a real-time (ReTi) PCR assay is described using a Taqman labeled probe for the detection and quantitation of infectious larygotracheitis virus (ILTV) in chickens. The ReTi ILTV assay was highly specific with a quantitation limit of 100 viral template copies per amplification reaction. In experimentally infected, birds during early acute stages of infection, an average of 6.67 log(10) viral template copies/amplification reaction were detected, while at chronic late stages of infection an average of 2.86-3.27 log(10) viral template copies/amplification reaction were detected. A total of 246 tracheal swab samples collected from natural outbreaks of the disease were tested by virus isolation and the ReTi ILTV assay. Both assays agreed in 37% of the samples tested and the ReTi ILTV assay detected approximately 3.7 times more positives samples than virus isolation. A minimum of 5 log(10) viral template copies/amplification reaction were required from a tracheal swab to render a virus isolation positive result. In conclusion, the ReTi ILTV assay was highly specific, sensitive, reproducible, and capable of reliably quantifying viral nucleic acid directly from clinical samples.     

Monastic nephrology in the School of Salerno and in an unpublished treatise in middlelatin and Italian volgare of the manuscript 2Qq C63 in the public library of Palermo

The relevance of the Salerno Medical School is examined through the contributions of Magister Maurus, Alfano I, Giovanni Plateario, Giovanni Afflacio, Bartolomeo and Urso of Calabria. An unpublished treatise on De Urinis in middlelatin and Italian volgare is introduced here for the first time by discussing the lapis of the ancient mediaeval urologic and uroscopic culture.     

The use of in situ hybridization and immunohistochemistry to study the pathogenesis of various Newcastle disease virus strains and recombinants in embryonated chicken eggs

Avian paramyxovirus type 1, commonly referred to as Newcastle disease virus (NDV), is a serious pathogen of significant economic importance to the industry. To investigate the role of the fusion (F), hemagglutinin-neuraminidase (HN), and (P) phosphoprotein gene sequences in virulence, six strains of Newcastle disease virus (NDV) representing all pathotypes and seven recombinant strains created by reverse genetics were inoculated into 9-day-old chicken embryos. Tissues and chorioallantoic membranes (CAM) were harvested at 24-hour intervals post-inoculation. Riboprobe in situ hybridization and immunohistochemistry highlighted distinct tissue tropisms among the viruses. Presence of F and/or HN from virulent viruses inserted into lentogenic backbones caused dissemination of virus in a manner similar to wild type virulent viruses. Disruption of P gene decreased dissemination of velogeinic infectious clones. It is concluded that each of these genes contributes to pathogenicity.     

Chemical composition and biological activity of a new type of Brazilian propolis: Red propolis

Propolis has been used as a medicinal agent to treat infections and promote wound healing for centuries. The aim of the present study was to test the antimicrobial, antioxidant, and cytotoxic activities of a new type of Brazilian propolis, popularly called red propolis, as well as to analyze its chemical composition. The antimicrobial activity against Staphylococcus aureus ATCC 25923 and Staphylococcus mutans UA159 was evaluated and the chloroform fraction (Chlo-fr) was the most active with lower MIC ranging from 25 to 50 μg/ml. The hexane fraction (H-fr), having the highest concentration of total flavonoids, showed the best sequestrating activity for the free radical DPPH. The ethanolic extract of propolis (EEP) showed cytotoxic activity for the HeLa tumor cells with an IC₅₀ of 7.45 μg/ml. When the EEP was analyzed by GC–MS, seven new compounds were found, among which four were isoflavones. Our results showed that the red propolis has biologically active compounds that had never been reported in other types of Brazilian propolis.     

Antimicrobial Susceptibility of Respiratory Isolates of Enterobacteriaceae and Staphylococcus aureus in Italy: Incidence and Trends Over the Period 1997–1999

The antibiotic susceptibility of members of the family Enterobacteriaceae and of Staphylococcus aureus strains isolated from the respiratory tract was assessed over the period 1997–1999 as part of the Italian Epidemiological Observatory survey sponsored by the SmithKline Foundation. A standardised method was used to determine the MICs of 22 antibiotics against isolates of Klebsiella pneumoniae (n=870), Escherichia coli (n=684), Enterobacter cloacae (n=342), Enterobacter aerogenes (n=187) and Serratia marcescens (n=135) as well as the MICs of 11 antibiotics against isolates of Staphylococcus aureus (n=1,606). Overall, the susceptibility rate of Enterobacteriaceae isolates was ≥90% to 5 agents (meropenem, imipenem, amikacin, cefepime and gentamicin); 89–80% to 2 agents (ciprofloxacin and tobramycin); and <80% to 11 agents (cefotaxime, piperacillin-tazobactam, trimethoprim-sulfamethoxazole, cefetamet, ceftriaxone, ceftazidime, aztreonam, ticarcillin-clavulanate, tetracycline, piperacillin, cefuroxime, chloramphenicol, ticarcillin, amoxicillin-clavulanate and amoxicillin). During the 3-year monitoring period, antibiotic susceptibility increased in Klebsiella pneumoniae against amoxicillin-clavulanate, in Escherichia coli against third-generation cephalosporins and aztreonam, in Enterobacter aerogenes against amoxicillin and piperacillin-tazobactam and in Serratia marcescens against most of the antibiotics. In contrast, Enterobacter cloacae showed a tendency to develop resistance to cefetamet, amikacin and ciprofloxacin. Of the total number of Staphylococcus aureus strains, 38% were methicillin resistant. Nearly 80% of the methicillin-resistant strains displayed a multiresistance pattern (additional resistance to 2 or more non-beta-lactam antibiotics). Rates of susceptibility of particular species (Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus) were compared using strains from different geographical areas of Italy (northern, central and southern) and from different nosocomial areas (outpatients, intensive care unit [ICU] inpatients, non-ICU inpatients). Susceptibility of Klebsiella pneumoniae to several antibiotics was lower in southern Italy, whereas the incidence of methicillin-resistant strains was higher in northern and central Italy. The susceptibility of Escherichia coli was similar in all three areas. No significant differences in susceptibility of Klebsiella pneumoniae or Escherichia coli were found between strains from inpatients and outpatients or from inpatients admitted to ICU and non-ICU units. The incidence of methicillin-resistant Staphylococcus aureus was higher in ICU inpatients (52%) than in non-ICU inpatients (38%) and lower in outpatients (19%) than in inpatients. Electronic Publication     

Tienilic acid in the treatment of arterial hypertension and congestive cardiac insufficiency

Tienilic acid (TA) is a common new diuretic agent with a potent uricosuric action. In a double-blind cross-over study its antihypertensive effect was compared to that of hydrochlorothiazide (HCT). 20 patients with essential hypertension were studied: after I weeks of placebo wash-out 10 patients received TA (dose range 250-750 mg/die) and 10 HCT (dose range 50-150 mg/die), for 5 weeks. Systolic and diastolic blood pressures were significantly and equally reduced (p < 0.001) after the first week of treatment in both groups. While serum uric acid concentration increased after HCT, it was significantly reduced (p < 0.001) after TA treatment. Serum potassium was slightly reduced with both treatments. Serum tryglicerides, unchanged after HCT, showed a slight tendency to reduction on TA treatment. Ten patients with congestive heart failure, on full digitalis treatment, were given TA (dose range 250-1000 mg/die): in each patient a prompt diuretic effect was observed, associated to a significant reduction of body weight and to a marked improvement of the clinical signs of heart failure. Therefore, TA is an effective diuretic agent which may be conveniently used in the treatment of arterial hypertension and congestive heart failure, as it induces a diuretic effect comparable to that obtained with HCT, reducing at the same time, serum uric acid levels.     

A new approach for calcitonin determination based on target cell responsiveness

We report the development and validation of three microbioassays for calcitonin based on calcitonin-induced inhibition of the activity of isolated osteoclasts. Having precisely quantified osteoclast motility, spreading and bone resorptive activity, we have applied stringent analytical procedures to define assay characteristics. We have found that the appropriately transformed responses significantly regress on log dose of the peptides. Furthermore, potency estimates obtained using calcitonins from three species (human, salmon and a synthetic analogue of eel calcitonin) have been found to be consistent with those obtained using conventional calcitonin bioassays. In addition, the assays are remarkably sensitive (detection limit 10(-15) M), highly specific and precise. We have determined plasma levels of bioactive calcitonin on samples from patients with medullary thyroid carcinoma; these are several-fold lower than those obtained using our routine calcitonin radioimmunoassay. Our study thus, forms the basis of an entirely new approach for the determination of 'biologically active' calcitonin, and we envisage that such target cell-specific assays could become useful microanalytical methods.