Clinical efficacy of nutraceutical diet for cats with clinical signs of cutaneus adverse food reaction (CAFR).

Food allergies and food intolerances are clinically difficult to discriminate. Most often, along with cutaneous adverse food reactions or CAFR, they are classified as adverse food reactions, whose causes are numerous, including toxic compounds. Eighteen indoor-housed domestic cats with evident clinical symptoms related to CAFR (drooling, back and neck intense itching, neck eczema, chronic conjunctivitis and stomatitis) involving skin lesions were studied. Cytological evaluations of ear, skin and gingival swabs revealed an increased turnover of keratinocytes while the oxytetracycline ELISA determination showed an unexpected high amount of oxytetracycline in all cats at the first visit. All cats were then randomly assigned to receive a standard (SD group) or a nutraceutical diet (ND group) for 60 days. In the ND group a significant reduction of the mean serum concentration of oxytetracycline, pruritus intensity and skin lesion severity (**p<0.01, ***p<0.001, and ***p<0.001, respectively) was observed after 60 days, and associated with a significant improvement in the clinical picture. Although a direct correlation between oxytetracycline presence within cat sera and CAFR-related symptoms has never been described, this study highlights the benefit of a specific nutraceutical diet supplementation in improving clinical symptoms and skin lesions in cats with CARF.

Reduction of peripheral blood T cells producing IFN-γ and IL-17 after therapy with abatacept for rheumatoid arthritis.

OBJECTIVES: Abatacept (ABA), a molecule used in the treatment of rheumatoid arthritis (RA), competes with the engagement of CD28, a T-cell receptor for co-stimulatory signals. CD28-mediated signalling regulates several T-cell functions, including inflammatory cytokine production and regulatory T cells (Treg) differentiation. Therefore, our objective was to evaluate the effects of ABA on peripheral blood T-lymphocyte cytokine production and on the number of circulating Treg. METHODS: In 24 RA patients treated with ABA for at least 6 months the proportions and absolute numbers of peripheral blood T cells producing interferon-gamma (IFN-γ) and interleukin-17 (IL-17) after in vitro stimulation, as well as those of Treg were longitudinally evaluated by flow cytometry. RESULTS: At baseline, compared with 16 healthy controls, RA patients had a higher percentage of CD4+ and CD8+ T cells producing IL-17 (p=0.021, and p=0.006, respectively), as well as of circulating Treg (p=0.041). After 6 months of therapy with ABA, there was a decrease of the percentage of IFN-γ- and IL-17-producing CD8+ T cells (p=0.033 and p=0.035, respectively), and of Treg (p=0.008), while that of IL-17-producing CD4+ T cells decreased after 12 months of treatment (p=0.005). The number of IL-17-producing T cells and of Treg, higher than in controls at baseline, normalised after ABA therapy. All these variations were statistically significant only in RA patients with EULAR good clinical response (n=17). CONCLUSIONS: The blockade of CD28 signal caused by ABA induces the decrease in peripheral blood of IL-17- and IFN-γ-producing T cells.

α-costic acid, the main sesquiterpenoid isolated from Dittrichia viscosa (L) Greuter, induces oxidative stress and autophagy in tomato.

Dittrichia viscosa (L.) Greuter, a perennial plant in the Asteraceae, has strong allelopathic activity due to the high content of various secondary metabolites. The bicyclic sesquiterpenoid α-costic acid is the most abundant secondary metabolite of D. viscosa. Its remarkable insecticidal, antiparasitic, and phytotoxic activities point to its potential use as a natural herbicide, but information on its mode of action is lacking. To shed light on the mechanism of action of α-costic acid in plant cells, we investigated the phytotoxicity of α-costic acid in tomato plants (Solanum lycopersicum L.) through in vivo assays, the underlying cellular effects using biochemical assays, and the effect on subcellular organelles using confocal microscopy on tomato protoplasts incubated with organelle-specific fluorescent probes. In vivo tests showed that α-costic acid inhibited the growth of tomato seedlings and induced chlorosis and spot lesions in leaves. Biochemical assays demonstrated that α-costic acid caused ion leakage, chlorophyll depletion, H2O2 overproduction, callose deposition, and membrane lipid peroxidation. Confocal microscopy demonstrated that α-costic acid determined ROS overproduction and network disruption in mitochondria, singlet oxygen overproduction in chloroplasts, vacuole disintegration, and autophagosome formation. Overall, our data are consistent with a model according to which α-costic acid phytotoxicity is related to oxidative stress in mitochondria and chloroplasts that induces extensive membrane damage, ultimately resulting in cell death associated with autophagy.

Health care costs of complex perianal fistula in Crohn's disease.

AIM: To evaluate the use of health care resources and the associated costs of complex perianal Crohn's disease (CD) from the National Health System perspective. METHODS: We conducted a multicenter, retrospective, observational study in which gastroenterologists from 11 hospitals in the Community of Madrid took part. Data was collected on the direct healthcare resources (pharmacological treatments, surgical procedures, laboratory/diagnostic tests, visits to specialists and emergency departments, and hospitalizations) consumed by 97 adult patients with complex perianal CD which was active at some point between January 1, 2005, and case history review. RESULTS: We recorded 527 treatments: 73.1% pharmacological (32.3% antibiotic, 20.5% immunomodulator, 20.3% biological) and 26.9% surgical. Mean annual global cost was €8,289/patient, 75.3% (€6,242) of which was accounted for by pharmacological treatments (€13.44 antibiotics; €1,136 immunomodulators; €5,093 biological agents), 12.4% (€1,027) by hospitalizations and surgery, 7.7% (€640) by medical visits, 4.2% (€350) by laboratory/diagnostic tests, and 0.4% (€30) by emergency department visits. CONCLUSIONS: Pharmacological therapies, and in particular biological agents, are the main cost driver in complex perianal CD; costs due to surgery and hospitalizations are much lower.

Modulation of the central memory and Tr1-like regulatory T cells in multiple sclerosis patients responsive to interferon-beta therapy.

BACKGROUND: Interferon-beta is used to reduce disease activity in multiple sclerosis, but its action is incompletely understood, individual treatment response varies among patients, and biological markers predicting clinical benefits have yet to be identified. Since it is known that multiple sclerosis patients have a deficit of the regulatory T-cell subsets, we investigated whether interferon-beta therapy induced modifications of the two main categories of regulatory T cells (Tregs), natural and IL-10-secreting inducible Tr1 subset, in patients who are biologically responsive to the therapy. METHODS: T-cell phenotype was determined by flow cytometry, while real-time PCR was used to evaluate interferon-beta bioactivity through MxA determination, and to measure the RNA for IL-10 and CD46 molecule in peripheral blood mononuclear cells stimulated with anti-CD46 and anti-CD3 monoclonal antibodies, which are known to expand a Tr1-like population. RESULTS: Interferon-beta induced a redistribution of natural Treg subsets with a shift of naive Tregs towards the 'central memory-like' Treg population that expresses the CCR7 molecule required for the in vivo suppressive activity. Furthermore, in a subgroup of treated patients, the CD46/CD3 co-stimulation, probably through the Tr1-like subset modulation, increased the production of RNA for IL-10 and CD46. The same group showed a lower median EDSS score after two years of therapy. CONCLUSIONS: The selective increase of 'central memory-like' subset and the involvement of the Tr1-like population may be two of the mechanisms by which interferon-beta achieves its beneficial effects. The quantification of RNA for IL-10 and CD46 could be used to identify patients with a different response to interferon-beta therapy.

Renewal of the T-cell compartment in multiple sclerosis patients treated with glatiramer acetate.

The immunomodulating activity of glatiramer acetate on T-cells of multiple sclerosis patients has only been partially clarified. The objective of this work was to investigate whether glatiramer acetate modifies thymic release of newly produced T-cells and the peripheral composition of the T-cell repertoire. T-cell receptor excision circles, (thymic) naive (CD4(+)CD45RA(+)CCR7(+)CD31(+)) T helper cells, and central (CD4(+)CD45RA(-)CCR7(+)) and effector (CD4(+)CD45RA(-)CCR7(-)) memory T-cells were evaluated in 89 untreated patients, 84 patients treated for at least 1 year, and 31 patients beginning treatment at the time of inclusion in the study and then followed-up for 12 months; controls were 81 healthy donors. The T-cell repertoire was analysed in selected samples. The percentage of (thymic)naive T helper cells was diminished in untreated patients, but rose to control values in treated subjects; a decrease in central memory T-cells was also observed in treated patients. Follow-up patients could be divided into two subgroups, one showing unmodified (thymic)naive T helper cells and T-cell diversity, the other in which the increased release of new T-cells was accompanied by modifications of the T-cell repertoire. Glatiramer acetate modifies the peripheral T-cell pool by activating a thymopoietic pathway of T-cell release that leads to a different setting of T-cell diversity and, likely, to a dilution of autoreactive T-cells.

Adult-type hypolactasia genotyping in Northern Italy: prevalence of C/T-13910 polymorphism and questions after comparison with existing data.

A genotyping assay was setup to assess the prevalence, in the population of a Northern Italian city, of the C/T-13910 single nucleotide polymorphism, closely associated to lactose malabsorption in many world areas including Sardinia. The results were compared to published Italian data, in order to evaluate the worth of a future validation of the assay for use in routine practice. DNA was extracted from blood samples of 123 randomly chosen healthy blood donors coming from the same city area, and was analyzed by a real-time polymerase chain reaction (PCR) genotyping assay; the frequency of the hypolactasia-associated CC-genotype was compared to the weighted average of results extracted from studies reporting the frequency of hypolactasic phenotype or genotype in nearby or distant Italian regions. Sixty-five percent of donors carried the CC-genotype, a percentage similar to other northern Italian cities, but significantly higher than what previously determined in surrounding Italian regions at the phenotype level, i.e. by breath test. This discrepancy parallels recent reports of non concordance between results of genotyping and hypolactasic phenotype in some world areas, including a neighbouring Northern Italian city. A north-south gradient of CC-prevalence was also observed. These results reinforce the notion of wide inter-regional variations in the frequency of C/T-13910 polymorphism and of incostant concordance with hypolactasic phenotype, even in subjects from the same country. Given the unsatisfactory results recently obtained from validation of a related assay in a neighbouring city, the authors decided not to proceed further and keep the assay only as a diagnostic aid in special situations.

Burkholderia gladioli sinonasal infection.

INTRODUCTION: Burkholderia gladioli are non-fermenting, Gram-negative, rod-shaped aerobic bacteria that were first identified as a plant pathogen. Most of the B. gladioli infections reported in the literature have involved immunocompromised adults and newborn infants. B. gladioli in humans is often associated with a poor prognosis. CASE REPORT: We describe the first case of sinonasal infection due to B. gladioli and Staphylococcus aureus in an immunocompetent patient who had recently travelled to the Congo. DISCUSSION: As in the few other reported cases involving immunocompetent patients, the appropriate approach to this multidrug-resistant B. gladioli infection was a combination of surgery and antibiotics chosen in the light of an antibiogram.

Groundwater and surface water quality characterization through positive matrix factorization combined with GIS approach.

This study aims at testing the effectiveness of Positive Matrix Factorization in characterizing groundwater and surface water quality, in terms of identifying main hydrochemical features and processes (natural and anthropogenic) that govern them. This method is applied in a hydro-system featured by a strong interrelation between groundwater and surface water and highly impacted by agricultural activities. Therefore, a holistic approach considering groundwater together with the surface water bodies, consisting in lake, several rivers and springs, was used. Multivariate statistical analysis, in particular Factor Analysis, has been proved to be effective in elaborating and interpreting water quality data highlighting the information carried within them, but it presents some limitations: it does not consider data uncertainty and it groups variables which are correlated positively and negatively. Moreover, in some cases the resulting factors are not clearly interpretable, describing each one various overlapping features/processes. Here, Positive Matrix Factorization is applied to groundwater and surface water quality data, and the results are compared to those obtained through a Factor Analysis in terms of both factor profiles and their spatial distribution through a GIS approach. Results of isotopes analysis are used to validate PMF output and support interpretation. Positive Matrix Factorization allows to consider data uncertainty and the solution respects two positivity constraints, based on the concept of chemical mass balance, which leads to a more environmentally interpretable solution. Results show that Positive Matrix Factorization identifies five different factors reflecting main features and natural and anthropogenic processes affecting the study area: 1) surface water used for irrigation, 2) groundwater subjected to reducing processes at advanced stages, 3) groundwater subjected to reducing processes at early stages, 4) groundwater residence time and 5) the effects of the agricultural land use on both groundwater and surface water. Positive Matrix Factorization leads to a more detailed understanding of the studied system as compared to Factor Analysis which identifies only three factors with overlapping information. Based on the results of this study, Positive Matrix Factorization could be a useful technique to perform groundwater and surface water quality characterization and to reach a deeper understanding of the phenomena that govern water chemistry.

Cytokines and osteolysis around total hip prostheses.

The aim of this work is to assess the correlation between the osteolysis around the prosthesis and the presence of cytokines favouring inflammation in the tissues at the interface between loosened prosthesis and bone. In this study, twenty-nine patients that underwent revision surgery were examined. Bioptic samples were collected at the interface between bone and implant both at the stem and socket level. Semiquantitative immunohistochemistry was performed to detect interleukin 1 alpha, interleukin 1 beta, interleukin 6 and tumour necrosis factor, cytokines that directly cause bone resorption and indirectly induce synthesis of other bone resorbing cytokines. Wear particles were identified and quantified by light microscopy. Radiographic evidence for osteolysis was scored by the Engh and Bobyn score. In tissues collected at the interface, the percentage of cells positive to IL1, IL6 and particularly to TNF increased in relation to the tissues collected at the interface with stable components. The cells occurring in the new capsule do not secrete cytokines in quantities that can be related to severity of wear. Cemented prostheses showed higher incidence of severe osteolysis, and higher levels of cytokines. It can be concluded that TNF, and to a lesser extent IL1 and IL6, are positively related to the severity of osteolysis around the prosthesis and therefore a pharmacological treatment can be hypothesized with anti-inflammatory or anti-cytokine drugs in order to limit or to avoid prosthesis loosening.

Utilización del termómetro timpánico en el consultorio pediatrico

Evalua la utilidad clínica de la termometría timpánica en 50 pacientes pediatricos con enfermedad febril con temperatura >38§c visto en un consultorio pediatrico privado entre noviembre 1994 y marzo 1995. 28 pacientes fueron <3 años(56 por ciento); y 22 pacientes fueron >3 años (44 por ciento). A todos los pacientes se les midió la temperatura timpánica en ambos oidos con el termómetro timpánico FIRST TEMP en siu modo oral. A los niños <3 años se les midió la temperatura rectal y a los niños >3 años se les midió la temperatura oral por 2 minutos con el termometro de mercurio. La temperatura promedio timpánica fue de 38,39§c, la oral de 38,19§c, y la rectal de 39,01§c. Esta diferencia fue estadisticamente significativa(p=<0,01). Existió una correlación entre la temperatura oral y timpánica(r= <0,68), una correlación entre la temperatura rectal y timpánica(r= 0,82). El termómetro timpánico tuvo una correlación con el termómetro de mercurio para la medición de la temperatura corporal en pacientes pediatricos con enfermedad febril. Su uso en el consultorio fue rápido, eficaz y bien tolerado por los pacientes