M. Vitale

Molecular cloning of the mouse CCK gene: expression in different brain regions and during cortical development

In this paper we describe experiments that address specific issues concerning the regulation of the mouse cholecystokinin gene in brain and intestine. The mouse cholecystokinin gene was cloned and sequenced. Extensive homology among the mouse, man and rat genes was noted particularly in the three exons and the regions upstream of the RNA start site. RNAse protection assays for each of the three exons were used to demonstrate that CCK is expressed in only a subset of tissues and that the same cap site and splice choices are used in brain, intestine as well as in cerebellum, cortex, midbrain, hypothalamus and hippocampus. CCK RNA was also noted to be detectable in kidney. Thus the same gene using the same promoter is expressed in subsets of cells that differ in their biochemical, morphologic and functional characteristics. The level of expression of CCK was also monitored during mouse cortical development and the appearance of CCK RNA was compared to glutamate decarboxylase (GAD), enkephalin and somatostatin. It was noted that each of these cortical markers was first expressed at different times during cortical development. The appearance of CCK RNA during intestinal development was also measured and found to precede appearance in cortex by several days.

Prion protein gene frequencies in three Sicilian dairy sheep populations

Abstract The objective of this paper was to investigate the prion protein (PrP) genotype and haplotype frequencies in three Sicilian dairy sheep populations. The three populations were: (1) 1096 Valle del Belice animals, (2) 1143 Comisana animals, and (3) 1771 individuals from 5 flocks with scrapie outbreaks, in which the animals were crossbreds derived from indigenous Sicilian dairy breeds. PrP genotypes are described for the three codons 136 (Alanine or Valine; A, V), 154 (Histidine or Arginine; H, R), and 171 (Glutamine, Arginine or Histidine; Q, R, H) which represent polymorphisms known to be linked with scrapie susceptibility. The Valle del Belice haplotype frequencies were 32.3% ARR, 6.5% AHQ, 1.0% ARH, 58.8% ARQ, and 1.4% VRQ. The Comisana frequencies were 39.4% ARR, 2.9% AHQ, 2.9% ARH, 50.9% ARQ, and 3.9% VRQ. In the flocks with scrapie outbreaks the frequencies were 32.8% ARR, 2.4% AHQ, 1.7% ARH, 59.1% ARQ, and 3.9% VRQ. In all three populations ARQ and ARR were the most frequent haplotypes. Multiple generations of strong selection will be needed to fixate the most resistant ARR haplotype.

Transcription in bacteriophage f1-infected Escherichia coli: RNA synthesized on DNA of deletion mutant PII shows the existence of a two-site terminator

SummaryTwo different transcripts are synthesized on the DNA of deletion mutant PII of bacteriophage f1 in E. coli cells infected with this miniphage. Both RNA species appear to be primary transcripts and differ by about 100 nucleotides at their 3OH end. Mapping of these molecules on the miniphage genome suggests that a two-site terminator is active at the end of the I region of transcription of bacteriophage f1.

M. bovis infection in pigs: Improvement of the γ-IFN assay efficiency in this species using a maintenance medium

The interferon-gamma (IFN-γ) test measures cell mediated immune response (CMI) during the early stages of tuberculosis infection. Although Bovine Tuberculosis (BT) spread in feral pigs is widely documented in literature, the effectiveness of IFN-γ in this species has been only recently reported. One of the major obstacle of this assay is that whole blood samples should be stimulated with purified protein derivative (PPD) cocktail within 8xa0h from the blood sampling. This study set up a defined broth culture in which lymphocytes, the cell population predominantly responsible for IFN-γ production, are maintained in a steady-state and their vitality is preserved. The IFN-γ production measured from the samples added with the maintenance medium and stored at 4xa0°C was similar to the enzyme-linked immunosorbent assay (ELISA) optical density values obtained from the same assay performed within 8xa0h from sampling.

Lesions Consistent with Tuberculous Spondylitis in Domestic and Wild Swine and Their Potential Use as a Model for Pott Disease in Humans

During a routine abattoir inspection of pig carcasses and control activities of hunted wildlife in 2013, 118 large white pigs Sus scrofa domesticus, 474 Nebrodi black pigs and 135 wild boars Sus scrofa scrofa were submitted to anatomopathological examination to evaluate the presence of tuberculosis-like lesions. Localized and generalized granulomatous lesions were detected with a prevalence of about 25% in large white pigs, 13% in Nebrodi black pigs and 8.15% in wild boars. Localized lesions involved mainly the submandibular lymph nodes, but when the disease was spread throughout the body, the inner organs and, also, in some cases, udders and/or bones were injured. The highest prevalence of generalized lesions (15/30) was observed in large white pigs, the only ones in which tuberculous granuloma affected also the spine. The bovine tuberculous spondylitis cases observed showed some similarities with Pott disease in humans regarding aspect and localization of lesions and age of the affected animals.Tissue samples of the positive animals were collected and submitted to bacteriological analysis, and the bone samples were also subjected to histological and immunohistochemistry analysis. M. bovis was isolated in all the analysed samples, and the granuloma encapsulation was found often incomplete indicating that the disease was in an active phase. The presence of lesions associated to tuberculous spondylitis in pigs suggests the possibility to use this animal species as model for the study of Pott disease in humans.

Analysis of differences in prion protein gene (PRNP) polymorphisms between Algerian and Southern Italy's goats

Abstract Goats are adapted to live in harsh areas of the world ensuring minimum levels of milk and meat production without much economic input. Scrapie is a fatal prion disease of small ruminants detected in different countries worldwide. Polymorphisms in the prion protein gene (PRNP) modify the degree of susceptibility/resistance to scrapie. PRNP polymorphisms were analysed in the four main Algerian goat breeds (Naine de Kabylie, Arbia, Mozabite, Mekatia), and in two native breeds of South Italy (Aspromontana from Calabria and Cilentana from Campania). The results were compared also with two previously described Sicilian native breeds (Girgentana and Rossa Mediterranea). Seven amino acid substitutions were detected in Naine de Kabylie goat which represents the original native Berber breed. All other breeds presented no more than six variants and common polymorphisms were present at codons, 154 and 240. Isoleucine at 137 was present in Algerian breeds only. The Italian Cilentana shared more variants with Algerian breeds whereas the Aspromontana breed was the only one to show a Serine at 127. Exclusive new variants were not detected since all polymorphisms were already described in other goats worldwide. The scrapie protective allele encoding lysine (K) at codon 222 was detected in the Naine de Kabylie and M’zabite breeds at low-frequency whereas is present with frequencies higher than 10% in all the Italian breeds reared in regions with elevated scrapie incidence. The overall results showed a substantial number of polymorphisms in PRNP, particularly in Naine de Kabylie breed, which carried also unique genotypes.

Pedigree verification with multiple candidate parents using Pedverif software

Abstract Species like sheep and beef cattle are commonly raised in large herds and often on pasture with multiple sires joining the females for unrecorded natural insemination. This leads to offspring with multiple candidate parents and therefore uncertain parentage. Twins or triplets can be from multiple sires as well. Pedigrees from such populations are often problematic and need proper verification. The Pedverif computer program is able to verify normal pedigrees as well as pedigrees with multiple candidate parents.

Rho-dependence of the terminator active at the end of the I region of transcription of bacteriophage f1

SummaryInfection of rho-Escherichia coli cells with deletion mutant PII of bacteriophage f1 results in a miniphage RNA population composed of transcripts longer than those synthesized in the infection of rho+ cells. This indicates a Rho dependence of the terminator active at the end of the I region of transcription of bacteriophage f1.An estimate of the length of a transcript, which represents a good fraction of the RNA that passes beyond the terminator, indicates that the hairpin structure where synthesis of complementary strand DNA initiates also acts as a fairly efficient Rho-independent terminator.