Genetic analysis of seven Italian horse breeds based on mitochondrial DNA D-loop variation.

To understand the origin and genetic diversity of Italian horses, mitochondrial DNA D-loop sequences were generated for 163 horses from seven breeds. Sequence analysis of a 480-bp segment revealed a total of 84 haplotypes with 57 polymorphic sites, indicating multiple maternal origins and high genetic diversity. Comparison of the haplotypes with the equine mtDNA haplotype/haplogroup nomenclature showed a haplogroup distribution in the Italian breeds more similar to that found in the Middle East breeds than in the European breeds, probably due to the economic and cultural relationship with the Middle East in the past centuries.

Detecting population structure and recent demographic history in endangered livestock breeds: the case of the Italian autochthonous donkeys.

Since its domestication, about 5000 years ago, the donkey (Equus asinus) has been extensively used as a work or draft animal in agricultural activities and for the transportation of people and goods. In the last century, technology improvement and growing mechanization strongly affected agriculture and the management and use of this livestock species in the industrialized countries. Nowadays, the use of donkeys for work or transport has almost disappeared, together with the need for mules or hinny breeding. During the last five decades, Italian autochthonous donkey populations suffered from a severe reduction in population size, which led to the extinction of several breeds. At present, eight breeds remain, all classified by FAO as critically endangered or endangered: Asinara, Pantesco, Grigio Siciliano, Romagnolo, Amiatino, Sardo Grigio, Martina Franca, and Ragusano. To evaluate the extant genetic variability of Italian donkeys, we typed 16 microsatellite loci in 258 individuals from these breeds. The results highlighted moderate levels of inbreeding ( F (IS) = 0.127) and a significant partition of genetic variation into breeds, as suggested by fixation index ( F (ST) = 0.109) and analysis of molecular variance (10.86% of total variation assigned to the between-breeds level) analyses. This was confirmed by a Bayesian clustering procedure that also highlighted a further partitioning at lower hierarchical levels corresponding to the farms of origin. This evidence suggests that an effective management strategy for Italian donkey populations should focus on breeds as conservation units. However, this requires a synergic management strategy at the farm level to maintain diversity and avoid inbreeding.

Derechos sexuales y reproductivos en la atención de adolescentes: diagnóstico de cumplimiento del marco legal / Sexual and reproductive rights in adolescents care: diagnosis of legal framework compliance

INTRODUCCIÓN Los avances en la garantía de los derechos sexuales y reproductivos han provocado cambios en las prácticas profesionales a favor de los usuarios y usuarias del sistema de salud. La población de adolescentes emerge como un colectivo particular y focalizado en el cual el cumplimiento de sus derechos los convierte en sujetos legítimos de prácticas responsables, centradas en el aspecto positivo de la salud. En la ciudad de Buenos Aires existe un conjunto de leyes que garantizan el pleno ejercicio de los derechos sexuales y reproductivos. Los profesionales que trabajan con adolescentes deben conocer e incorporar este marco legal al abordaje de la salud integral. OBJETIVOS Analizar el conocimiento y la implementación del marco legal que garantiza los derechos sexuales y reproductivos en la atención de adolescentes en dos hospitales públicos, que forman parte del Programa de Salud Integral del Adolescente del Gobierno de la Ciudad de Buenos Aires. MÉTODOS Se realizó un estudio descriptivo cualitativo durante 2013-14 en tres etapas consecutivas. En la primera se analizó el marco legal; la segunda permitió definir las dimensiones y los ámbitos de indagación; en la última etapa se efectuaron las entrevistas a profesionales en distintos ámbitos de atención (internación, consultorio y guardia). RESULTADOS Según los resultados del estudio, la mayoría de los profesionales sabe que la ley garantiza la información, prescripción y provisión de los métodos anticonceptivos, pero sólo la mitad realiza esas prestaciones en la consulta con adolescentes. Casi todos conocen y prescribirían la anticoncepción hormonal de emergencia, pero en la práctica menos de la mitad la entrega. DISCUSIÓN Existen diferencias en la aplicación del marco legal. Se observa un abordaje integral sólo en el ámbito ambulatorio y cuando hay intervención de profesionales no médicos. La salud sexual y reproductiva no se aborda durante la internación de adolescentes. En guardia, la anticoncepción hormonal de emergencia se entrega solamente en obstetricia.

Cold affects the transcription of fatty acid desaturases and oil quality in the fruit of Olea europaea L. genotypes with different cold hardiness.

The olive tree lacks dormancy and is low temperature sensitive, with differences in cold tolerance and oil quality among genotypes. The oil is produced in the drupe, and the unsaturated fatty acids contribute to its quality. The aim of the present research was to investigate the relationship among development, cold response, expression of fatty acid desaturase (FAD) genes, and unsaturated fatty acid composition in drupes belonging to genotypes differing in leaf cold tolerance, but producing good oil (i.e. the non-hardy Moraiolo, the semi-hardy Frantoio, and the hardy Canino). In all genotypes, cold sensitivity, evaluated by cold-induced transient increases in cytosolic calcium, was high in the epi-mesocarp cells before oil body formation, and decreased during oil biogenesis. However, genotype-dependent differences in cold sensitivity appeared at the end of oil production. Genotype-dependent differences in FAD2.1, FAD2.2, FAD6, and FAD7 expression levels occurred in the epi-mesocarp cells during the oleogenic period. However, FAD2.1 and FAD7 were always the highest in the first part of this period. FAD2.2 and FAD7 increased after cold applications during oleogenesis, independently of the genotype. Unsaturated fatty acids increased in the drupes of the non-hardy genotype, but not in those of the hardy one, after cold exposure at the time of the highest FAD transcription. The results show a direct relationship between FAD expression and lipid desaturation in the drupes of the cold-sensitive genotype, and an inverse relationship in those of the cold-resistant genotype, suggesting that drupe cold acclimation requires a fine FAD post-transcriptional regulation. Hypotheses relating FAD desaturation to storage and membrane lipids, and genotype cold hardiness are discussed.

Genetic diversity and admixture analysis of Sanfratellano and three other Italian horse breeds assessed by microsatellite markers.

Sanfratellano is a native Sicilian horse breed, mainly reared in the north east of the Island, developed in the 19th century from local dams and sires with a restricted introgression of Oriental, African and, more recently, Maremmano stallions. In this study, the genetic relationships and admixture among Sanfratellano, the other two Sicilian autochthonous breeds and Maremmano breed were assessed using a set of microsatellites. The main goals were to infer the impact of Maremmano breed in the current Sanfratellano horse and to provide genetic information useful to improve the selection strategies of the Sanfratellano horse. The whole sample included 384 horses (238 Sanfratellano, 50 Sicilian Oriental Purebred, 30 Sicilian Indigenous and 66 Maremmano), chosen avoiding closely related animals. A total of 111 alleles from 11 microsatellite loci were detected, from four at HTG7 to 15 at ASB2 locus. The mean number of alleles was the lowest in Oriental Purebred (6.7), the highest in Sanfratellano (8.3). All the breeds showed a high level of gene diversity (He) ranging from 0.71 ± 0.04 in Sicilian Oriental Purebred to 0.81 ± 0.02 in Sicilian Indigenous. The genetic differentiation index was low; only about 6% of the diversity was found among breeds. Nei's standards (DS) and Reynolds' (DR) genetic distances reproduced the same population ranking. Individual genetic distances and admixture analysis revealed that: (a) nowadays Maremmano breed does not significantly influence the current Sanfratellano breed; (b) within Sanfratellano breed, it is possible to distinguish two well-defined groups with different proportions of Indigenous blood.

An international parentage and identification panel for the domestic cat (Felis catus).

Seventeen commercial and research laboratories participated in two comparison tests under the auspices of the International Society for Animal Genetics to develop an internationally tested, microsatellite-based parentage and identification panel for the domestic cat (Felis catus). Genetic marker selection was based on the polymorphism information content and allele ranges from seven random-bred populations (n = 261) from the USA, Europe and Brazil and eight breeds (n = 200) from the USA. Nineteen microsatellite markers were included in the comparison test and genotyped across the samples. Based on robustness and efficiency, nine autosomal microsatellite markers were ultimately selected as a single multiplex 'core' panel for cat identification and parentage testing. Most markers contained dinucleotide repeats. In addition to the autosomal markers, the panel included two gender-specific markers, amelogenin and zinc-finger XY, which produced genotypes for both the X and Y chromosomes. This international cat parentage and identification panel has a power of exclusion comparable to panels used in other species, ranging from 90.08% to 99.79% across breeds and 99.47% to 99.87% in random-bred cat populations.

Genomics of berry fruits antioxidant components.

Reactive oxygen and nitrogen metabolites, which are side products of cell metabolism, can produce a lot of damage in biological macromolecules and tissues, producing a number of chronic illnesses. On the other hand, antioxidant metabolites usually accumulated in fruits and vegetables can provide an effective protection by neutralizing these reactive molecules. Among comestible vegetables, berry fruits are considered one of the richest sources of antioxidant metabolites; hence, they represent a good model for molecular and biochemical investigations about the biosynthesis and the functional role of antioxidants in plants. This review illustrates how recent developments in the fields of genomics and bioinformatics can provide powerful tools to better understand the molecular mechanisms that trigger biosynthesis and accumulation of antioxidant metabolites in berries.

Italian Group for Lap-Band System: results of multicenter study on patients with BMI < or =35 kg/m2.

BACKGROUND: The Lap-Band System is the most common bariatric operation world-wide. Current selection criteria do not include patients with BMI < or = 35. We report the Italian multicentre experience with BMI < or = 35 kg/m(2) over the last 5 years. PATIENTS AND METHODS: Data were obtained from 27 centres involved in the Italian Collaborative Study Group for Lap-Band System. Detailed information was collected on a specially created electronic data sheet (MS Access 2000) on patients operated in Italy since January 1996. Items regarding patients with BMI < or = 35 were selected. Data were expressed as mean +/- SD except as otherwise indicated. RESULTS: 225 (6.8%) out of 3,319 Lap-Band patients were recruited from the data-base. 15 patients, previously submitted to another bariatric procedure (BIB =14; VBG= 1) were excluded. 210 patients were eligible for study (34M/176F, mean age 38.19+/-11.8, range 17-66 years, mean BMI 33.9+/-1.1, range 25.1-35 kg/m(2), mean excess weight 29.5+/-7.1, range 8-41). 199 comorbidities were diagnosed preoperatively in 55/210 patients (26.2%). 1 patient (0.4%) (35 F) died 20 months postoperatively from sepsis following perforation of dilated gastric pouch. There were no conversions to laparotomy. Postoperative complications presented in 17/210 patients (8.1%). Follow-up was obtained at 6, 12, 24, 36, 48 and 60 months. At these time periods, mean BMI was 31.1+/-2.15, 29.7+/-2.19, 28.7+/-3.8, 26.7+/-4.3, 27.9+/-3.2, and 28.2+/-0.9 kg/m(2) respectively. Co-morbidities completely resolved 1 year postoperatively in 49/55 patients (89.1%). At 60 months follow-up, only 1 patient (0.4%) has a BMI >30. CONCLUSIONS: Although surgical indications for BMI < or = 35 remain questionable, the Lap-Band in this study demonstrated that all but 1 patient achieved normal weight, and most lost their co-morbidities with a very low mortality rate.

The Italian Group for LAP-BAND: predictive value of initial body mass index for weight loss after 5 years of follow-up.

BACKGROUND: Laparoscopic of the LAP-BAND System placement stage of obesity is a safe operation, but its indication in terms of stage of obesity is controversial. The aim of this study was to evaluate the 5 years stage of obesity results for weight loss in patients with varying preoperative ranges of body mass index (BMI). METHODS: Data were obtained from the Italian Collaborative Study Group for LAP-BAND System (GILB) registry. Detailed information was collected on a specifically created database (MS Access 2000) for patients operated on in Italy from January 1996 to 2003. Patients operated on between January 1996 and December 1997 were allocated to four groups according to preoperative BMI range: 30-39.9 kg/m(2) (group A), 40-49.9 kg/m(2) (group B), 50-59.9 kg/m(2) (group C), and =60 kg/m(2) (group D) percent estimated weight loss respectively. Postoperative complications, mortality, BMI, BMI loss, and (%EWL) were considered in each group. Data are expressed as mean +/- SD, except as otherwise indicated. Statistical analysis was done by means of Fisher's exact test, and p < 0.05 was considered significant. RESULTS: After 5 years from LAP-BAND System surgery, 573 of 3,562 patients were eligible for the study. One hundred fifty-five of 573 (27.0%) were lost to follow-up, 24 of 418 (5.7%) underwent band removal due to complications (gastric pouch dilation, band erosion), eight of 418 (1.9%) were converted to other bariatric procedures, five of 418 (1.2%) died of causes not related to the operation or the band, and 381 of 573 (66.5%) were available for follow-up. Based on 96, 214, 64, and seven patients their preoperative BMI, Were allocated to groups A, B, C, and D, respectively. At time of follow-up mean BMI was 27.5 +/- 5.2 in group A, 31.6 +/- 4.7 in group B, 37.6 +/- 17.3 in group C, and 41.4 +/- 6.9 kg/m(2) in group D. Mean BMI loss was 9.8 +/- 5.4, 12.9 +/- 5.2, 15.8 +/- 8.1, and 23.2 +/- 4.9 kg/m(2), respectively, in groups A, B, C, and D. Mean %EWL at the same time was 54.6 +/- 32.3 in group A, 54.1 +/- 17.2 in group B, 51.6 +/- 35 in group C, and 59.l +/- 17.1 in group D. CONCLUSION: Initial BMI in this series did not correlate with %EWL 5 years after the operation. In fact %EWL was almost the same in each group, independent of preoperative weight. Initial BMI was an accurate indicator of the results obtained 5 years after LAP-BAND in group C (50-59.9 kg/m(2)) and D (=60 kg/m(2)) patients, who remained morbidly obese despite their %EWL.

Genetic dissection of blue-light sensing in tomato using mutants deficient in cryptochrome 1 and phytochromes A, B1 and B2.

Several novel allelic groups of tomato (Solanum lycopersicum L.) mutants with impaired photomorphogenesis have been identified after gamma-ray mutagenesis of phyA phyB1 double-mutant seed. Recessive mutants in one allelic group are characterized by retarded hook opening, increased hypocotyl elongation and reduced hypocotyl chlorophyll content under white light (WL). These mutants showed a specific impairment in response to blue light (BL) resulting from lesions in the gene encoding the BL receptor cryptochrome 1 (cry1). Phytochrome A and cry1 are identified as the major photoreceptors mediating BL-induced de-etiolation in tomato, and act under low and high irradiances, respectively. Phytochromes B1 and B2 also contribute to BL sensing, and the relative contribution of each of these four photoreceptors differs according to the light conditions and the specific process examined. Development of the phyA phyB1 phyB2 cry1 quadruple mutant under WL is severely impaired, and seedlings die before flowering. The quadruple mutant is essentially blind to BL, but experiments employing simultaneous irradiation with BL and red light suggest that an additional non-phytochrome photoreceptor may be active under short daily BL exposures. In addition to effects on de-etiolation, cry1 is active in older, WL-grown plants, and influences stem elongation, apical dominance, and the chlorophyll content of leaves and fruit. These results provide the first mutant-based characterization of cry1 in a plant species other than Arabidopsis.

Tomato contains homologues of Arabidopsis cryptochromes 1 and 2.

Cryptochromes are blue light photoreceptors found in both plants and animals. They probably evolved from photolyases, which are blue/UV-light-absorbing photoreceptors involved in DNA repair. In seed plants, two different cryptochrome (CRY) genes have been found in Arabidopsis and one in Sinapis, while three genes have been found in the fern Adiantum. We report the characterisation of tomato CRY genes CRY1 and CRY2. They map to chromosomes 4 and 9, respectively, show relatively constitutive expression and encode proteins of 679 and 635 amino acids, respectively. These proteins show higher similarity to their Arabidopsis counterparts than to each other, suggesting that duplication between CRY1 and CRY2 is an ancient event in the evolution of seed plants. The seed plant cryptochromes form a group distinct from the fern cryptochromes, implying that only one gene was present in the common ancestor between these two groups of plants. Most intron positions in CRY genes from plants and ferns are highly conserved. Tomato cryl and cry2 proteins carry C-terminal domains 210 and 160 amino acids long, respectively. Several conserved motifs are found in these domains, some of which are common to both types of cryptochromes, while others are cryptochrome-type-specific.

Outbreak of a Leaf Disease Caused by Pseudocercospora ceratoniae on Carob in Sicily.

Carob (Ceratonia siliqua L.), an evergreen tree typical of the Mediterranean flora, has been grown in Sicily (Italy) from time immemorial for its fruits, used mainly as food for cattle and horses as well as for industrial production of alcohol. Although the economic importance of carob as a commercial crop has declined over the last decades, carob trees still are a characteristic aspect of the landscape in southeastern Sicily. In early April 1998, a severe outbreak of a foliar disease was noted on carob trees in the Ragusa province. Symptoms initially consisted of small (2 to 3 mm wide) dark brown, vein-limited spots, visible on both sides of the leaf and, later in the season, surrounded by a pale halo. In a humid atmosphere, spots scattered over the leaf blade but usually were most numerous along the midrib, enlarged, and coalesced, forming large blotches. Severely affected leaflets dropped, leaving the petiole attached to the tree. As a result, the trees appeared defoliated. Severely defoliated trees did not produce fruits. The causal agent of this disease was identified as Pseudocercospora ceratoniae (Pat. & Trab.) Deighton (1), an hyphomycetous fungus reported previously as a pathogen of carob under the name Cercospora ceratoniae Pat. & Trab. (3). On carob leaflets, P. ceratoniae formed grayish caespituli, confined to the lower surface of the necrotic spots. Caespituli consisted of dense fascicles of conidiophores (up to more than 50 conidiophores per fascicle) emerging through the stomata. Conidiophores were simple, slightly ampulliform, and geniculate at the conidial scars, which were conspicuous and unthickened. Old scars often were situated laterally on a short denticle. Conidia, borne singly as terminal blastospores and varying considerably in length, were pluriseptate, filiform, substraight or slightly curved, frequently slightly obclavate, with an obtuse apex and a short constriction at the base toward the truncate, unthickened hilum. Conidia from pure cultures of the fungus grown on water agar under black light were suspended in water and sprayed onto pot-grown carob plants. Inoculated plants were kept in a moist chamber for 48 h and subsequently transferred to the greenhouse. After 12 to 14 days leaf spots similar to those observed on naturally infected trees developed on inoculated plants and the pathogen was reisolated. Control plants sprayed with distilled water remained symptomless. C. ceratoniae had been recorded previously on carob in various Mediterranean countries, including Italy (2), but since has attracted little attention, being regarded as a sporadically occurring pathogen. Both mild temperatures during the winter and exceptionally frequent and persistent rain during the spring may have favored the epidemic outbreak of the disease caused by this fungus. References: (1) F. C. Deighton. 1976. Mycol. Pap. No. 140. Commonw. Mycol. Inst., Kew, England. (2) R. Parisi. Boll. Orto Bot. R. Univ. Napoli 10:155, 1932. (3) L. Roger. 1953. Phytopathologie des Pays Chauds. Vol. 2. P. Lechevalier, Paris.

Reduced requirement for RNA editing in the mitochondrial cox3 transcript of Olea europaea L.

Transcripts from the mitochondrial cox3 locus in Olea europaea L. are edited in ten nucleotide positions. Nine of these C-to-U transitions affect 3.4% of the genomically encoded amino-acid identity to specify a COXIII polypeptide better conserved in evolution. RNA editing of cox3 in olive tree mitochondria is, thus, less extensive than in the other higher plants so far investigated. This low RNA editing frequency might correlate with both the GC content observed in the cox3 gene and the phylogenetic position of Olea.

RNA editing in the cox3 mRNA of Magnolia is more extensive than in other dicot or monocot plants.

The Magnoliaceae are discussed as one of the key species at the root of the flowering plants. To obtain molecular information for one of these phylogenetically interesting plant species, we determined genomic and cDNA sequences of the mitochondrial cox3 gene in Magnolia grandiflora. Twenty-two RNA editing events are identified to alter cytidines in the mRNA to uridines, all but one of which change the encoded amino acid identity. RNA editing in the cox3 coding region is thus more frequent in Magnolia than in other dicot or monocot plants investigated and almost as predominant as in some gymnosperms. The cox3 RNA editing frequency in Magnolia thus occupies an intermediate position between angiosperms and gymnosperms consistent with the phylogenetic position of the Magnoliales.

Conservation of the organization of the mitochondrial nad3 and rps12 genes in evolutionarily distant angiosperms.

The organization of the genes nad3 and rps12 has been investigated in the mitochondrial genome of two dicotyledonous plants - Helianthus and Magnolia - and one monocotyledonous plant (Allium). These plants all contain a complete rps12 gene downstream of the nad3 gene. This arrangement is thus highly conserved within angiosperms. The two genes are co-transcribed and the transcript is modified at several positions by RNA editing of the C to U-type, thus confirming that both genes encode functional proteins. Some 26, 35 and 27 editing events have been identified in the PCR-derived nad3-rps12 cDNA population from sunflower, Magnolia and onion, respectively. Editing of the nad3-rps12 transcript is thus more extensive in Magnolia than in the other angiosperms so far investigated and radically changes the genomically encoded polypeptide sequence. A novel species-specific codon modification was observed in Magnolia. Several homologous sites show differences in editing pattern among plant species. A C-to-U alteration is also found in the non-coding region separating the nad3 and rps12 genes in sunflower. The PCR-derived cDNA populations from the nad3-rps12 loci analysed were found to be differently edited. In addition the plant species show marked variations in the completeness of RNA editing, with only the Magnolia nad3 mRNA being edited fully.

Comparison of the Abbott Cell Dyn 3000 SL and the Coulter STKS hematology analyzers.

A side by side comparison of the Cell Dyn 3000 SL and Coulter STKS using identical samples was performed over a four month period in the Clinical Hematology Laboratory at the University of Cincinnati Medical Center. A total of 444 samples comprised of 114 sophomore medical students and 330 randomly selected clinical patients were compared in 20 hemogram parameters, and the ability of each instrument to generate differential suspect flags was analyzed. Correlation coefficients for leukocytes, erythrocytes, hemoglobin, hematocrit, and platelets were 0.99. Correlation coefficients for the Cell Dyn and STKS compared with a 600 cell manual differential were 0.98 and 0.89 for neutrophils, 0.96 and 0.87 for lymphocytes, and 0.72 and 0.48 for monocytes, respectively. Both instruments demonstrated high precision and accuracy by internal and external quality control standards. Each analyzer exhibited strength as a screening instrument for abnormal cell populations. The Coulter STKS had overall sensitivity of 75%, specificity of 94%, 25% false negatives, and 6% false positives. Sensitivity, specificity, false negative and positive rates for the Cell Dyn 3000 SL were 68%, 92%, 32%, and 8%, respectively. Based upon this extensive side by side comparison using an identical sample population, it has been concluded that although statistically significant systematic bias (p < 0.05) exists between the two instruments, both analyzers can adequately support the needs of the clinical hematology laboratory.

[Acute pancreatitis in biliary microlithiasis: a study of the biliary sediment]. / Pancreatite acuta nella microlitiasi biliare: studio del sedimento biliare.

The Authors report the results of a clinical trial based on the study of the biliary sediment in cases of acute pancreatitis considered as idiopathic that, on the contrary, resulted to be correlated to a condition of biliary microlithiasis. Four cases of biliary acute pancreatitis out of 94 are reported (1979-1988) in which the diagnosis of biliary microlithiasis was possible only through the microscopic analysis of the bile, that documented a significant presence of cholesterol and/or bilirubinate crystals. It allowed a proper therapeutic approach.

Monitoring of antifibrinolytic therapy following subarachnoid hemorrhage. The importance of CSF fibrin/fibrinogen degradation products.

Ten cases of subarachnoid hemorrhage (SAH) from ruptured cerebral aneurysm are reported. Fibrin/fibrinogen degradation product (FDP) levels were determined simultaneously in blood and cerebrospinal fluid (CSF) at an average frequency of 1.7 days, extended over periods of 8 to 63 days. Successful antifibrinolytic therapy (AFT) correlated with FDP levels in CSF of less than 16 micrograms/ml. Five patients failed to respond to AFT. Levels of FDP in the CSF fluctuated widely in these five patients, and remained at or above 16 micrograms/ml for most of the monitoring period. Blood FDP levels were normal or minimally elevated, and could not be used in predicting or preventing rebleeding episodes. A hypothesis is presented to explain the significance of the presence of FDP's in CSF. In spite of the many techniques employed in monitoring AFT and reviewed in this paper, little information has been gained to improve the results and therapeutic strategies. Among the different methods available, FDP measurements in the CSF have correlated best with rebleeding, and thus may be used in modifying and individualizing therapy. Suggestions are given for future studies.