Angelo Peli

The role of the peroxisome proliferator-activated receptor-α (PPAR-α) in the regulation of acute inflammation

The peroxisome proliferator‐activated receptor‐α (PPAR‐α) is a member of the nuclear receptor superfamily of ligand‐dependent transcription factors related to retinoid, steroid, and thyroid hormone receptors. The aim of the present study was to evaluate the role of the PPAR‐α receptor on the development of acute inflammation. To address this question, we used two animal models of acute inflammation (carrageenan‐induced paw edema and carrageenan‐induced pleurisy). We report here that when compared with PPAR‐α wild‐type mice, PPAR‐α knockout mice (PPAR‐αKO) mice experienced a higher rate of the extent and severity when subjected to carrageenan injection in the paw edema model or to carrageenan administration in the pleurisy model. In particular, the absence of a functional PPAR‐α gene in PPAR‐αKO mice resulted in a significant augmentation of various inflammatory parameters (e.g., enhancement of paw edema, pleural exudate formation, mononuclear cell infiltration, and histological injury) in vivo. Furthermore, the absence of a functional PPAR‐α gene enhanced the staining (immunohistochemistry) for FAS ligand in the paw and in the lung and the expression of tumor necrosis factor α and interleukin‐1β in the lungs of carrageenan‐treated mice. In conclusion, the increased inflammatory response observed in PPAR‐αΚΟ mice strongly suggests that a PPAR‐α pathway modulates the degree of acute inflammation in the mice.

Palmitoylethanolamide and luteolin ameliorate development of arthritis caused by injection of collagen type II in mice

IntroductionN-palmitoylethanolamine (PEA) is an endogenous fatty acid amide belonging to the family of the N-acylethanolamines (NAEs). Recently, several studies demonstrated that PEA is an important analgesic, antiinflammatory, and neuroprotective mediator. The aim of this study was to investigate the effect of co-ultramicronized PEA + luteolin formulation on the modulation of the inflammatory response in mice subjected to collagen-induced arthritis (CIA).MethodsCIA was induced by an intradermally injection of 100 μl of the emulsion (containing 100 μg of bovine type II collagen (CII)) and complete Freund adjuvant (CFA) at the base of the tail. On day 21, a second injection of CII in CFA was administered. Mice subjected to CIA were administered PEA (10 mg/kg 10% ethanol, intraperitoneally (i.p.)) or co-ultramicronized PEA + luteolin (1 mg/kg, i.p.) every 24 hours, starting from day 25 to 35.ResultsMice developed erosive hind-paw arthritis when immunized with CII in CFA. Macroscopic clinical evidence of CIA first appeared as periarticular erythema and edema in the hindpaws. The incidence of CIA was 100% by day 28 in the CII-challenged mice, and the severity of CIA progressed over a 35-day period with a resorption of bone. The histopathology of CIA included erosion of the cartilage at the joint. Treatment with PEA or PEA + luteolin ameliorated the clinical signs at days 26 to 35 and improved histologic status in the joint and paw. The degree of oxidative and nitrosative damage was significantly reduced in PEA + luteolin-treated mice, as indicated by nitrotyrosine and malondialdehyde (MDA) levels. Plasma levels of the proinflammatory cytokines and chemokines were significantly reduced by PEA + luteolin treatment.ConclusionsWe demonstrated that PEA co-ultramicronized with luteolin exerts an antiinflammatory effect during chronic inflammation and ameliorates CIA.

Effects of a polyphenol present in olive oil, oleuropein aglycone, in a murine model of intestinal ischemia/reperfusion injury.

Dietary olive oil supplementation and more recently, olive oil phenols have been recommended as important therapeutic interventions in preventive medicine. Ole has several pharmacological properties, including antioxidant, anti‐inflammatory, antiatherogenic, anticancer, antimicrobial, and antiviral and for these reasons, is becoming an important subject of study in recent years. The aim of this study was to investigate the effects of Ole aglycone on the modulation of the secondary events in mice subjected to intestinal IRI. This was induced in mice by clamping the superior mesenteric artery and the celiac trunk for 30 min, followed by release of the clamp, allowing reperfusion for 1 h. After 60 min of reperfusion, animals were killed for histological examination of the ileum tissue and immunohistochemical localization of proinflammatory cytokines (TNF‐α and IL‐1β) and adhesion molecules (ICAM‐1 and P‐sel); moreover, by Western blot analysis, we investigated the activation of NF‐κB and IκBα. In addition, we evaluated the apoptosis process, as shown by TUNEL staining and Bax/Bcl‐2 expressions. The results obtained by the histological and molecular examinations showed in Ole aglycone‐treated mice, a decrease of inflammation and apoptosis pathway versus SAO‐shocked mice. In conclusion, we propose that the olive oil compounds, in particular, the Ole aglycone, could represent a possible treatment against secondary events of intestinal IRI.

Evaluation of CALC-I gene (CALCA) expression in tissues of dogs with signs of the systemic inflammatory response syndrome.

Objective – To perform a qualitative evaluation of procalcitonin gene (CALCA) expression in a tissue-specific manner in dogs with signs of the systemic inflammatory response syndrome (SIRS). Design – Observational study. Setting – University Veterinary Teaching Hospital. Animals – Nine clinical cases and 5 research dogs. Interventions – None. Measurements and Main Results – Fresh tissue samples (thyroid, lung, liver, spleen) from 9 dogs that died with a diagnosis of parvoviral infection or SIRS were collected and immediately stored at –80°C. Diagnosis of parvoviral infection was based on clinical signs, positive fecal antigen test, and confirmed by polymerase chain reaction (PCR). Clinical diagnosis of SIRS was based on the clinical criteria reported in veterinary literature. Necropsy was performed on all subjects in the study. Furthermore, thyroid, lung, liver, spleen were collected from 5 normal research dogs immediately postmortem for testing. The 9 dogs with a clinical diagnosis of SIRS died from either parvovirus (n=5), bacterial sepsis (n=3), or neoplasia (n=1). CALCA was amplified by PCR in the following samples: thyroid (9/9), spleen (6/9), lung (4/9), liver (3/9). Only thyroid expressed CALCA in the 5 normal dogs. Conclusions – In SIRS, extrathyroidal transcription of CALCA was documented. Quantitative analysis (real-time polymerase chain reaction) in a wider population of SIRS and normal dogs will provide essential information about the extent and source of extrathyroidal expression of canine CALCA induced by septic and nonseptic systemic inflammation.OBJECTIVE To perform a qualitative evaluation of procalcitonin gene (CALCA) expression in a tissue-specific manner in dogs with signs of the systemic inflammatory response syndrome (SIRS). DESIGN Observational study. SETTING University Veterinary Teaching Hospital. ANIMALS Nine clinical cases and 5 research dogs. INTERVENTIONS None. MEASUREMENTS AND MAIN RESULTS Fresh tissue samples (thyroid, lung, liver, spleen) from 9 dogs that died with a diagnosis of parvoviral infection or SIRS were collected and immediately stored at -80 °C. Diagnosis of parvoviral infection was based on clinical signs, positive fecal antigen test, and confirmed by polymerase chain reaction (PCR). Clinical diagnosis of SIRS was based on the clinical criteria reported in veterinary literature. Necropsy was performed on all subjects in the study. Furthermore, thyroid, lung, liver, spleen were collected from 5 normal research dogs immediately postmortem for testing. The 9 dogs with a clinical diagnosis of SIRS died from either parvovirus (n=5), bacterial sepsis (n=3), or neoplasia (n=1). CALCA was amplified by PCR in the following samples: thyroid (9/9), spleen (6/9), lung (4/9), liver (3/9). Only thyroid expressed CALCA in the 5 normal dogs. CONCLUSIONS In SIRS, extrathyroidal transcription of CALCA was documented. Quantitative analysis (real-time polymerase chain reaction) in a wider population of SIRS and normal dogs will provide essential information about the extent and source of extrathyroidal expression of canine CALCA induced by septic and nonseptic systemic inflammation.

The effects of different environmental conditions on thermoregulation and clinical and hematological variables in long-distance road-transported calves

The aim of this study was to investigate the effects of long-distance road transport (19 h, from Poland to Italy) during 2 seasons (summer vs. winter) on clinical and hematological variables in calves. The environmental temperature range that could compromise the thermoregulation system (thermal stress) of the calves was tested. For the 7 Holstein calves in each transport, the BW and rectal temperature (RT) were measured, and blood samples were collected at the farm of origin, before loading at the transit center (T2), after unloading at the farm of destination (T3), and 1, 2, 3, and 4 d after arrival. The body temperature (BT) and heart rate (HR) were continuously monitored from T2 to T3. The data were statistically analyzed according to a mixed model that considered the fixed effects of transport (repeated measurements), season of journey, and their interaction. Within the observed temperature-humidity index (THI) range (30 to 80), effective thermoregulation allowed the calves to maintain their BT with small physiologic changes to prevent thermal stress, particularly in the summer. With no seasonal differences, the HR was greater at loading than unloading (120 vs. 115 beats per min; P = 0.012). As for the transport effect, the BW was less (P < 0.001) after unloading, and the RT was greater (P = 0.004). This effect was more marked in summer. The hematological variables indicated a moderate effect of transport on the hydration condition, reactive and muscular systems, and metabolism, although hematocrit (P = 0.004), erythrocytes, cortisol, NEFA, β-hydroxybutyrate, lactate, creatine kinase, lactate dehydrogenase, and aspartate aminotransferase activity (P < 0.001), and total protein (P = 0.007) were greater after unloading. This was confirmed by a moderate decrease in total leukocytes (P = 0.031) and glucose concentration (P = 0.002). The changes in the clinical variables were similar for both seasons even though in the summer, hematocrit (P < 0.001), urea (P = 0.008), and total protein (P = 0.010) increased and glucose concentration (P = 0.038) decreased. In conclusion, the data did not show a pronounced effect attributable to the season of the journey. Long-distance road transport leads to notable changes in clinical and hematological variables at the end of the journey. However, these variables remained within their physiological ranges and returned to basal values within a few days after the journey.

Expression of interleukin-1β, interleukin-8, and interferon-γ in blood samples obtained from healthy and sick neonatal foals

OBJECTIVE To evaluate and compare the gene expression of interleukin(IL)-1β, IL-8, and interferon-γ during the first 72 hours after birth in healthy foals and during the first 72 hours after hospitalization in sick neonatal foals and investigate correlations of clinicopathologic variables with cytokine expressions in healthy and sick neonatal foals. ANIMALS 33 foals < 7 days old (10 healthy foals, 7 foals with sepsis, 6 foals with peripartum asphyxia syndrome, and 12 foals with other diseases [2 with failure of passive transfer of immunity only were not further evaluated]). PROCEDURES A blood sample (15 mL) was collected from each foal immediately after birth or hospital admission (0 hours) and at 24 and 72 hours later. Clinicopathologic variables were evaluated, and cytokine gene expression in WBCs was measured with an absolute quantitative real-time reverse transcriptase PCR assay. RESULTS At all time points, gene expression of interferon-γ was low in all groups. No time-dependent changes in cytokine expressions were detected in healthy or sick foals. Foals with sepsis had significantly higher IL-1β gene expression than did healthy foals, foals with peripartum asphyxia syndrome, or foals with other diseases. At 0 hours, IL-1β expression was correlated with plasma fibrinogen concentration in healthy foals and with the neutrophil-to-lymphocyte ratio in foals with sepsis; IL-8 expression was correlated with monocyte count in foals with sepsis and with arterial pH, plasma fibrinogen concentration, and plasma lactate concentration in foals with peripartum asphyxia syndrome. CONCLUSIONS AND CLINICAL RELEVANCE Data have suggested that evaluation of IL-1β expression in sick neonatal foals could help identify those with sepsis.

Four-Year Monitoring of Foodborne Pathogens in Raw Milk Sold by Vending Machines in Italy

Prevalence data were collected from official microbiological records monitoring four selected foodborne pathogens (Salmonella, Listeria monocytogenes, Escherichia coli O157:H7, and Campylobacter jejuni) in raw milk sold by self-service vending machines in seven Italian regions (60,907 samples from 1,239 vending machines) from 2008 to 2011. Data from samples analyzed by both culture-based and real-time PCR methods were collected in one region. One hundred raw milk consumers in four regions were interviewed while purchasing raw milk from vending machines. One hundred seventy-eight of 60,907 samples were positive for one of the four foodborne pathogens investigated: 18 samples were positive for Salmonella, 83 for L. monocytogenes, 24 for E. coli O157:H7, and 53 for C. jejuni in the seven regions investigated. No significant differences in prevalence were found among regions, but a significant increase in C. jejuni prevalence was observed over the years of the study. A comparison of the two analysis methods revealed that real-time PCR was 2.71 to 9.40 times more sensitive than the culture-based method. Data on consumer habits revealed that some behaviors may enhance the risk of infection linked to raw milk consumption: 37% of consumers did not boil milk before consumption, 93% never used an insulated bag to transport raw milk home, and raw milk was consumed by children younger than 5 years of age. These results emphasize that end-product controls alone are not sufficient to guarantee an adequate level of consumer protection. The beta distribution of positive samples in this study and the data on raw milk consumer habits will be useful for the development of a national quantitative risk assessment of Salmonella, L. monocytogenes, E. coli O157, and C. jejuni infection associated with raw milk consumption.

Plasma concentrations and therapeutic effects of budesonide in dogs with inflammatory bowel disease.

OBJECTIVE To evaluate the pharmacokinetics and clinical efficacy of budesonide in dogs with inflammatory bowel disease (IBD). ANIMALS 11 dogs (mean ± SD age, 5.7 ± 3.9 years; various breeds and body weights) with moderate or severe IBD. PROCEDURES Each dog received a controlled-release formulation of budesonide (3 mg/m(2), PO, q 24 h) for 30 days (first day of administration was day 1). The concentration of budesonide and its metabolite (16-α-hydroxyprednisolone) was measured via liquid chromatography-tandem mass spectrometry in plasma and urine samples obtained on days 1 and 8 of treatment. On those days, plasma samples were obtained before the daily budesonide administration and 0.5, 1, 2, 4, and 7 hours after drug administration, whereas urine samples were obtained after collection of the last blood sample. A clinical evaluation was performed on the dogs before onset of drug administration and on days 20 and 30 after start of drug administration. RESULTS The highest plasma concentration of budesonide and 16-α-hydroxyprednisolone on day 1 was detected at 1 hour and at 2 hours after drug administration, respectively. After standardization on the basis of specific gravity, the ratio between urinary concentrations of budesonide and 16-α-hydroxyprednisolone was 0.006 and 0.012 on days 1 and 8, respectively. The clinical response was adequate in 8 of 11 dogs. CONCLUSIONS AND CLINICAL RELEVANCE Budesonide was rapidly absorbed and metabolized in dogs with IBD. The drug gradually accumulated, and there was an adequate therapeutic response and no adverse effects.

Cervus elaphus papillomavirus (CePV1): New insights on viral evolution in deer

We identified a novel papillomavirus (CePV1) in a fibropapilloma of a 1.5 year old male red deer (Cervus elaphus) shot in the Italian Alps in Brescia province. PV particles were first observed by electron microscopy and PV DNA was then identified by PCR using degenerate primers. Subsequently we cloned the entire genome and determined its complete sequence. CePV1 genome is 8009 bp long and contains all 9 ORFs and the long untranslated regulatory region characteristic for Delta-papillomaviruses. Pairwise nucleotide alignments and phylogenetic analyses based on concatenated E1-E2-L1 ORFs allowed to determine the highest similarity with the Capreolus caprelus papillomavirus CcaPV1. The analysis of the host-parasite phylogenetic tree interactions suggest the co-divergence of CePV1 and C. elaphus while the identified topological incongruences leading us to speculate that CcaPV1 could eventually be the result of an earlier host switch event.

Equine Bronchoalveolar Lavage Cytokines in the Development of Recurrent Airway Obstruction

Pietra, M., Peli, A., Bonato, A., Ducci, A. and Cinotti, S., 2007. Equine bronchoalveolar lavage cytokines in the development of recurrent airway obstruction. Veterinary Research Communications, 31(Suppl. 1), 313–316