Relationship between alcoholism addiction and periodontitis. An in vivo study using drinking-in-darkness protocol in rats.

Periodontal disease (PD) has been considered a probable risk factor for several systemic diseases. Among them, PD is presumed to be one of the possible etiologies of chronic illness of the central nervous system. In this context, poor oral health and PD is associated with substance abuse in humans. However, if periodontal lesions can produce addiction is unknown. This paper aims to evaluate the possibility that chronic periodontal injury (CPL) can cause ethanol binge intake in drink-in-darkness (DID) protocol in rats. In CPL group (n=10) experimental damage was done to the periodontal tissue of the secondmaxillary molar, the control group (n=9) received sham injury. Forty-three days after CPL the intake of ethanol was assessed using several concentrations in DID experiment. During the DID experiment, we observed significant differences between the binge-type consumption of ethanol at the lowest concentration of 10% (p=0.01). Differences in consumption of 20% ethanol are observed during a few days (p=0.04), and there are no differences in consumption at 40% concentration of ethanol (p=0.2). It is concluded that chronic periodontal lesion leads to alcoholism in Wistar rats

Molecular diagnosis of Theileria and Babesia species infecting cattle in Northern Spain using reverse line blot macroarrays

BACKGROUND: Piroplasmosis in cattle is caused by tick-borne haemoprotozoan parasites of the genera Theileria and Babesia. Molecular detection techniques offer higher sensitivity and specificity than microscopy examination methods and serological tests. A reverse line blot (RLB) macroarray that included generic and species-specific probes for Theileria annulata, Theileria buffeli, Babesia bovis, Babesia bigemina, Babesia divergens and Babesia major was used to study the presence and identity of the piroplasm species infecting 263 bovine blood samples from 79 farms, most of them in Northern Spain. Microscopy examination of blood smears and haematology were also performed whenever possible to identify animals with parasitaemia. RESULTS: RLB hybridisation identified infection in 54.0% of the samples, whereas only 28.8% were positive by microscopy examination. The most frequently found species was T. buffeli, present in 42.6% of the samples. T. annulata was found in 22 samples (8.4%) from 12 farms, including 9 farms (14 samples) located in Northern Spain where presence of the vector is not very common. Babesia infections were less frequently detected: B. major was found in 3.0% of the samples, B. bigemina in 2.7%, B. bovis in 2.3% and B. divergens in 1.1%. Mixed infections were detected in 14 samples, accounting for six different combinations of species. CONCLUSION: This is the first report in which B. major and B. divergens have been detected in Spain using molecular identification techniques and the first time that B. bovis has been detected in Northern Spain. The detection of T. annulata in Northern Spain suggests that the distribution of Mediterranean theileriosis might be changing. Samples with positive RLB hybridisation but negative microscopy had haematology values within the normal ranges suggesting that they corresponded to chronic carriers that may serve as reservoirs of the infection. In this sense, sensitive and specific laboratorial tests like RLB that clearly identify the parasite and can detect subclinical infections are essential to establish good control measures

Assessment of exposure to piroplasms in sheep grazing in communal mountain pastures by using a multiplex DNA bead-based suspension array

BACKGROUND: Piroplasms are tick-borne hemoprotozoans with a major impact on extensive management systems. Detection of sub-clinical low-level carriers, which can act as source of infection for vector ticks, is key to protect livestock trade and facilitate preventive control programs. The purpose of this study was to develop a method for the detection of ovine piroplasms and to use it in a field study aimed at investigating piroplasms infection in semi-extensive production systems in the Basque Country (northern Spain). METHODS: A DNA bead-based suspension array using the Luminex® xMAP technology that included a generic Theileria-Babesia control probe, 6 species-specific probes, and an internal control probe was developed to detect and identify piroplasms that infect sheep. To monitor piroplasm infection in clinically healthy sheep from 4 flocks that share communal mountain pastures, blood samples were collected during 2 grazing seasons. RESULTS: Piroplasms were detected in 48% (214/446) of blood samples, nearly half of them (49.1%, 105/214) as mixed infections. Five different piroplasms were identified: Theileria sp. OT3 in 34.8% of the samples, Theileria ovis in 20.9%, and at lower prevalences Babesia motasi (12.3%), Theileria luwenshuni/OT1 (10.5%) and Babesia ovis (6.3%). Despite differences among flocks associated to differences in management, an increasing trend in the incidence of piroplasm infection with increasing age of animals after increased tick exposure was observed. This increment could be attributed to continued re-infection associated with re-exposure to ticks at grazing. Ticks were collected from animals (4 species) and vegetation (8 species), and associations between tick abundance seasonality and risk of infection with the different piroplasms were established. CONCLUSION: The multiplex Luminex® xMAP procedure is a rapid and high throughput technique that provided highly specific and sensitive identification of single and mixed piroplasm infections in blood of sheep carriers. This study confirmed a situation of endemic stability for piroplasm infection in the region, where infection is present in the absence of clinical signs, and mountain grazing allows for sufficient inoculation rates to maintain such situation

Expression of vascular endothelial growth factor in normal and tumoral liver mouse cells

In previous studies have been reported a controversy between the reactivity of mono and polyclonal antibodies in different cells populations of the mouse. In this study we focus on the immunoreactivity of the monoclonal antibody VEGF (C- 1) in order to analyze its expression in two cell populations of regenerating mouse liver (hepatocytes and endothelial cells) after partial hepatectomy, and two transplanted hepatocarcinomas (ES2 and SS1K). C3H/S male mice were divided into two groups and kept under standard conditions for circadian periodicity analysis, one group were transplanted with the tumors into the subcutaneous tissue of the animal flank and the other group were subjected to a partial hepatectomy (70 %). The animals were killed by decapitation and exsanguination. Samples of regeneration liver, ES2 and SS1K tumors were processed for histology and immunostained with VEGF (C-1) mouse monoclonal IgG2a antibody. The results showed strongly immunopositivity reaction for VEGF either in hepatocytes or endothelial cells of mouse regenerating liver. We also demonstrated positive immunoreaction in both malignant tumors ES2 and SS1K endothelial and parenchymal cells. We conclude that the use of VEGF (C-1) mouse monoclonal IgG2a antibody can be applied successfully in experimental design leading to study processes involved in mice’s angiogenesi

Analysis of the p16INK4 and TP53 Tumor Suppressor Genes in Bone Sarcoma Pediatric Patients

Recent data suggest that deletion of p16INK4 and mutation of TP53 are among the most common genetic events in the development of human cancer, since the codified proteins act as brakes of the abnormal cell cycle. As the molecular events leading to the development of pediatric bone sarcomas remain unclear, we analyzed 75 osteosarcoma and Ewing sarcoma samples from 43 pediatric patients to search for alterations at the TP53 or p16INK4 tumor suppressor genes. By means of PCR-DGGE (polymerase chain reaction and denaturing gradient gel electrophoresis) we detected TP53 point mutations in 18.6% of the tumor samples, but no constitutional mutations. In the analysis of p16INK4, 7% of the samples harbored deletions of the gene but no point mutations were detected by SSCP (single strand conformation polymorphism) analysis, just the polymorphism Ala-->Thr at codon 148. These data support the hypothesis that TP53 alterations may play a role in the development of pediatric bone tumors and that the primary mechanism of inactivation of p16INK4 seems to be homozygous deletion rather than point mutation

Estudio comparativo de la síntesis de ADN en hígado y riñón de ratones lactantes y adultos

In previous papers we have observed that the proliferative activity of numerous cell populations of suckling and young mice was different than the observed in adults. In the present work we analized the DNA synthesis of suckling and adult mice, by immunohistochemistry (Bromodioxiuridine) in both hepatocytes and renocytes, at two different time points. The object of the present experiment was to establish the correlation in cellular proliferation between both groups. We used C3HS suckling male mice of 21 days old and adult mice of 90 days old. The results showed that DNA synthesis of renocytes and hepatocytes in suckling mice were significantly higher than the adults, and we didn't find statistically differences between the values of both analysed time points. We can conclude that in suckling mice we could observe maximum and minimum values but we coudn't find a circadian rhythm like those established in adults.En trabajos previos hemos observado que la actividad proliferativa de numerosas poblaciones celulares del ratón lactante y joven difiere de la encontrada en el adulto. En el presente trabajo experimental, se estudia la síntesis de ADN del ratón lactante y adulto, mediante el método inmunohistoquímico de la Brdu (Bromodeoxiuridina), en las poblaciones celulares de hepatocitos y renocitos, a las OO:O0 h y 16:00 h, con el objeto de establecer la correspondencia en la proliferación celular entre ambos grupos etarios. Para ello se utilizaron machos lactantes de 21 días de edad y adultos de 90 días, de la cepa C3H/S. Los resultados muestran que los valores de síntesisde ADN de los renocitos y hepatodtos en los ratones lactantes son significativamente mayores (p<O.001) a los observados en los adultos, no encontrándose diferencias estadísticamente significativas entre los valores de ambos puntos horarios en los órganos estudiados. Podemos concluir que estos hallazgos demuestran que, si bien en los ratones lactantes aparecen valores máximos y mínimos en la actividad proliferativa de distintas poblaciones celulares, estas aún no presentan la ritmicidad establecida para los adultos

Expression of vascular endothelial growth factor in normal and tumoral liver mouse cells

In previous studies have been reported a controversy between the reactivity of mono and polyclonal antibodies in different cells populations of the mouse. In this study we focus on the immunoreactivity of the monoclonal antibody VEGF (C-1) in order to analyze its expression in two cell populations of regenerating mouse liver (hepatocytes and endothelial cells) after partial hepatectomy, and two transplanted hepatocarcinomas (ES2 and SS1K). C3H/S male mice were divided into two groups and kept under standard conditions for circadian periodicity analysis, one group were transplanted with the tumors into the subcutaneous tissue of the animal flank and the other group were subjected to a partial hepatectomy (70 %). The animals were killed by decapitation and exsanguination. Samples of regeneration liver, ES2 and SS1K tumors were processed for histology and immunostained with VEGF (C-1) mouse monoclonal IgG2a antibody. The results showed strongly immunopositivity reaction for VEGF either in hepatocytes or endothelial cells of mouse regenerating liver. We also demonstrated positive immunoreaction in both malignant tumors ES2 and SS1K endothelial and parenchymal cells. We conclude that the use of VEGF (C-1) mouse monoclonal IgG2a antibody can be applied successfully in experimental design leading to study processes involved in mice’s angiogenesisFacultad de Ciencias Médica

Spanish biofuels heating value estimation based on structural analysis

The importance of waste biomass as an energy source is likely to increase during the coming years as a result of European energy policy objectives, and because of the wide range of possibilities that it offers: it is a cheap fuel, widespread, and available in large quantities. In addition to crops and forestry operations, the Spanish fruit, olive and wine industries generate large amounts of currently undervalued solid wastes such as stones, branches, pulps or pomaces. The use of these by-products offers environmental benefits like removing waste and preventing fires at the same time as providing an energy yield. A proper energy valorization will require a complete physicochemical characterization. In this article, a structural and thermal characterization is developed from twenty samples from the olive and wine industries, as well as from forest and agro wastes. In addition, predictive equations are proposed to determine higher heating value (HHV) from chemical composition. For this purpose, the chemical extraction method (also called the ‘classic’ method) was used, and results were obtained in accordance with the data shown in the bibliography. Two predictive equations were developed: one based on lignin and hemicellulose content, and the other based on lignin quantity. Both present an absolute average error (AAE) of 0.87% and 1.13%, respectively.Beca predoctoral Severo Ochoa (BP13058) PSE-ARFRISOL (PS-120000-2005-1) MINECO (CTQ2013-45155-R