Intravarietal polymorphisms reveal possible common ancestor of native Schinus terebinthifolius Raddi populations in Brazil.

Schinus terebinthifolius Raddi is a perennial native from Atlantic forest. It is of high ecological plasticity and is used in traditional medicine. Based on promising reports concerning its bioactivity, it was included as a species of great interest for distribution through the National Health System. A number of agronomic studies to guide its crop production are therefore underway. This study examined diversity and phylogenetic relationships among native S. terebinthifolius populations from different Brazilian ecosystems: Cerrado; sandbanks; dense rainforest; and deciduous forest. The intergenic regions rpl20-5'rps12, trnH-psbA, and trnS-trnG were sequenced from cpDNA and aligned using BLASTn. There were few fragments for comparison in GenBank and so only region trnS-trnG was informative. There were variations among and within populations with intravarietal polymorphisms and three distinct haplotypes (HpSM, HpDDO, HpNE), once populations from NE (sandbanks and rainforest) clustered together. Sequences from HpSM, HpNE, and HpDDO returned greater similarity to haplotypes A (AY928398.1), B (AY928399.1), and C (AY928400.1), respectively. A network, built by median-joining among native haplotypes and 10 available on GenBank, revealed HpSM as the origin of all other haplogroups. HpDDO showed the most mutations and was closely related to haplogroups from Argentina. While this could indicate hybridization, we believe that the polymorphisms resulted from adaptation to events such as deforestation, fire, rising temperature, and seasonal drought during the transition from Atlantic forest to Cerrado. While more detailed phylogeographical studies are needed, these results indicate eligible groups for distinct climates as an important step for pre-breeding programs before field propagation.

Genome wide association study on early puberty in Bos indicus.

The aim of this study was to evaluate a genome wide association study (GWAS) approach to identify single nucleotide polymorphisms (SNPs) associated with fertility traits (early puberty) in Nellore cattle (Bos indicus). Fifty-five Nellore cows were selected from a herd monitored for early puberty onset (positive pregnancy at 18 months of age). Extremes of this phenotype were selected; 30 and 25 individuals were pregnant and non-pregnant, respectively, at that age. DNA samples were genotyped using a high-density SNP chip (>777.000 SNP). GWAS using a case-control strategy highlighted a number of significant markers based on their proximity with the Bonferroni correction line. Results indicated that chromosomes 5, 6, 9, 10, and 22 were associated with the traits of interest. The most significant SNPs on these chromosomes were rs133039577, rs110013280, rs134702839, rs109551605, and rs41639155. Candidate genes, as well as quantitative trait loci (QTL) previously reported in the Ensembl and Cattle QTLdb databases, were further investigated. Analysis of the regions close to the SNP on chromosomes 9 and 10 revealed that four QTL had been previously classified under the reproduction category. In conclusion, we have identified SNPs in close proximity to genes associated with reproductive traits. Moreover, U6 spliceosomal RNA was present on three different chromosomes, which is possibly associated with age at first calving, suggesting that it might be a strong candidate for future studies.

Genetic variations in the leptin gene associated with growth and carcass traits in Nellore cattle.

The aim of the study was to detect polymorphisms in the leptin gene and to determine the association of these polymorphisms with growth and carcass traits in Nellore cattle. The single nucleotide polymorphisms (SNPs) -1457 (AJ571671:g.-1457A>G) and A59V (AF536174.1:g. 321C>T), as well as the microsatellite BM1500 (3.9 kb downstream), were genotyped. The measures of body weight and ultrasound examinations (rib eye area, back, and rump fat thickness) were performed in 3 different periods of animal management. During the first period, the animals were fed with grass and mineralized salt ad libitum. In the second period, they received grass and concentrate, and in the third, only concentrate. After the slaughter of animals, data were collected for classification and typification of carcasses. No significant association was found between the variables assessed and SNP -1457. Conversely, SNP A59V was associated with rump fat thickness and muscle color post-slaughter. BM1500 was associated with rump fat thickness in the first period (pre-slaughter), subcutaneous fat thickness in the second, weight of the animals in the third, and length of the carcass after slaughter. These results suggest that SNP A59V and the microsatellite BM1500 might be useful for marker-assisted selection in Nellore cattle.

Oxigenación con membrana extracorpórea en la disfunción primaria del injerto en trasplante bipulmonar pediátrico: a propósito de un caso / Extracorporeal membrane oxygenation in primary graft dysfunction in a paediatric double lung transplant: presentation of a case

La disfunción primaria del injerto es la principal causa de morbimortalidad en el postoperatorio inmediato de los pacientes sometidos a trasplante pulmonar. Entre las opciones de tratamiento se encuentran las estrategias ventilatorias de protección pulmonar, el óxido nítrico, el surfactante pulmonar, así como medidas de soporte con dispositivo de membrana de oxigenación extracorpórea (ECMO), como puente a la recuperación de la función pulmonar o al retrasplante. Presentamos el caso de una niña de 9 años afectada de fibrosis quística y sometida a trasplante bipulmonar que en el postoperatorio inmediato presentó una disfunción primaria del injerto severa refractaria al tratamiento. Ante el desarrollo de disfunción multiorgánica se decidió el empleo de ECMO venoarterial (arteria pulmonar-aurícula derecha). La evolución posterior fue satisfactoria, permitiendo su retirada a los 5 días de postoperatorio. Actualmente la paciente vive libre de rechazo y con excelente calidad de vida tras 600 días de seguimiento (AU)

Primary graft dysfunction is a leading cause of morbimortality in the immediate postoperative period of patients undergoing lung transplantation. Among the treatment options are: lung protective ventilatory strategies, nitric oxide, lung surfactant therapy, and supportive treatment with extracorporeal membrane oxygenation (ECMO) as a bridge to recovery of lung function or re-transplant. We report the case of a 9-year-old girl affected by cystic fibrosis who underwent double-lung transplantation complicated with a severe primary graft dysfunction in the immediate postoperative period and refractory to standard therapies. Due to development of multiple organ failure, it was decided to insert arteriovenous ECMO catheters (pulmonary artery-right atrium). The postoperative course was satisfactory, allowing withdrawal of ECMO on the 5 th post-surgical day. Currently the patient survives free of rejection and with an excellent quality of life after 600 days of follow up (AU)

[Extracorporeal membrane oxygenation in primary graft dysfunction in a paediatric double lung transplant: presentation of a case]. / Oxigenación con membrana extracorpórea en la disfunción primaria del injerto en trasplante bipulmonar pediátrico: a propósito de un caso.

Primary graft dysfunction is a leading cause of morbimortality in the immediate postoperative period of patients undergoing lung transplantation. Among the treatment options are: lung protective ventilatory strategies, nitric oxide, lung surfactant therapy, and supportive treatment with extracorporeal membrane oxygenation (ECMO) as a bridge to recovery of lung function or re-transplant. We report the case of a 9-year-old girl affected by cystic fibrosis who underwent double-lung transplantation complicated with a severe primary graft dysfunction in the immediate postoperative period and refractory to standard therapies. Due to development of multiple organ failure, it was decided to insert arteriovenous ECMO catheters (pulmonary artery-right atrium). The postoperative course was satisfactory, allowing withdrawal of ECMO on the 5th post-surgical day. Currently the patient survives free of rejection and with an excellent quality of life after 600 days of follow up.

Genetic diversity of locally adapted sheep from Pantanal region of Mato Grosso do Sul.

Sheep of the Pantaneiro breed and seven other breeds, raised in the State of Mato Grosso do Sul, Brazil, were genotyped using eight microsatellite loci. The aim of the present study was to determine the genetic variability, phylogenetic relationship, and patterns of gene introgression and miscegenation among the animals surveyed, to obtain information about the genetic structure of locally adapted sheep in Mato Grosso do Sul. A total of 195 animals were used for genetic analysis. The Pantaneiro breed had the largest average number of alleles/locus (9.25), and higher allelic richness (6.95), while the Dorper population had the lowest values for these parameters (4.88 and 3.86, respectively). Analysis of genetic distance values and genetic structure between populations made it possible to characterize these animals with regard to distinct genetic groups. Average expected heterozygosity ranged from 0.72 (Pantaneiro) to 0.55 (Dorper), while average observed heterozygosity ranged from 0.63 (White Dorper) to 0.54 (Dorper). On the basis of the statistical parameters evaluated, it was possible to demonstrate that when compared to other populations, the Pantaneiro breed represented a reservoir of genetic diversity with rare and useful alleles for genetic improvement, emphasizing the importance of preserving the breed.

Recombinant AgB8/1 ELISA test vs. commercially available IgG ELISA test in the diagnosis of cystic echinococcosis.

The diagnosis and clinical management of cystic echinococcosis (CE) rely on imaging and serology, the latter still having a complementary role as its accuracy in assessing cyst viability is unsatisfactory. We used an experimental IgG ELISA test based on the recombinant antigen rEgAgB8/1 cloned from Echinococcus granulosus to differentiate active from inactive/cured CE infection, comparing its performance to that of a commercially available ELISA test used routinely in our hospital laboratory. Both tests were performed on sera from 88 patients with hepatic echinococcal cysts, grouped according to cyst stage based on ultrasonographical morphology, and on 17 patients surgically treated for echinococcosis and 18 patients with nonparasitic hepatic cysts included as controls. Tests' performances did not differ significantly, but the overall concordance between tests drastically dropped when groups were analysed separately. Further longitudinal studies should evaluate whether these discrepancies reflect the different ability of either test to predict the evolution of cysts over time. Although the recombinant-AgB8/1-based ELISA test seems to have no clinical advantage over the commercially available ELISA test in the assessment of hepatic CE cyst viability, the easiness of production and reproducibility of high-quality recombinant antigens makes rEgAgB8/1 a valid candidate for use in CE ELISA diagnostic tests.

Myostatin (GDF8) single nucleotide polymorphisms in Nellore cattle.

The myostatin gene, also known as GDF8 (growth differentiation factor 8), is located on bovine chromosome 2 (BTA2); it has three exons and two introns. Myostatin is specifically expressed during embryonic development and in adult skeletal muscle, functioning as a negative regulatory protein. Several cattle breeds (Piedmontese, Belgian Blue and Blond'Aquitaine, and others) show polymorphisms in this gene; these polymorphisms are directly related to the double muscling phenotype. We looked for polymorphisms in the Nellore cattle myostatin gene and compared them with those known for taurine breeds. Seven regions, covering the three exons of this gene, were amplified by polymerase chain reaction and sequenced, including the untranslated region. DNA from 30 adult Nellore animals was collected; DNA sequencing revealed three, seven and four polymorphisms in exons 1, 2 and 3, respectively. We found previously reported polymorphisms, as well as several new ones; for instance, 37 polymorphisms were found in the untranslated region segment, and in introns 1 and 2 there were one and three polymorphisms, respectively. The high degree of allelic heterogeneity in the myostatin gene could be related to its high mutation rate; it also could be the result of a long history of artificial selection for meat production, which has probably favored such modifications and maintained them in cattle populations. These polymorphisms identified in Nellore cattle could be useful for breeding programs.

Targeted nucleotide exchange in bovine myostatin gene.

The myostatin gene, known as Growth Differentiation Factor 8 (GDF8), located at chromosome 2 (BTA2) in cattle, is specifically expressed during embryo development and in the adult skeletal muscle. Molecular analysis of this gene reveals the presence of three exons and two introns. Several cattle breeds, such as Piedmontese, Belgian Blue, Blond'Aquitaine, among others, show polymorphisms in this gene, which are directly related to double muscling phenotype. Piedmontese cattle shows a nucleotide transition G --> A (G938A) at exon 3, resulting in the substitution of cysteine to tyrosine, leading to a protein structure change, which determines myostatin inactivation and consequent muscular hypertrophy. The objective of this work was to implant the polymorphism G938A, naturally existent in Piedmontese breed, into in vitro propagated foetal myoblasts, from Nellore cattle. Single strand DNA (ssDNA) oligonucleotides were used to direct the same nucleotidic transition (G938A) to exon 3. Two transfection protocols (cationic lipid solution and electroporation) were tested and, 48 hours after transfection, RNA and DNA were extracted from myoblasts. Reverse transcription and polymerase chain reaction (PCR) were performed, using primers flanking the mutation region. The PCR products were cloned and analyzed by DNA sequencing, and it was possible to detect the nucleotidic CT transition at position 938, in the electroporated myoblasts. The existence of a positive signal in the transfection indicates that ssDNA oligonucleotides can be used to introduce this point mutation in specific functional gene sites.

A simple sonographic scoring system combined with routine serology is useful in differentiating parasitic from non-parasitic cysts of the liver().

In the absence of a detached endocyst, unilocular echinococcal cysts of the liver may be difficult to distinguish from non-parasitic cysts. In an attempt to identify sonographic features that could help distinguish these two types of cysts, we retrospectively analyzed 64 cases of fluid-filled hepatic cysts whose parasitic nature was ultimately excluded. This experience allowed us to develop a simple scoring system that quantifies the likelihood that hepatic cysts are non-parasitic. Sonographic criteria, together with the results of standard serological testing for cystic echinococcosis, proved to be sufficiently specific to allow definitive diagnosis without resorting to further tests.

Genetic diversity of microsatellite loci in Leopardus pardalis, Leopardus wiedii and Leopardus tigrinus.

The microsatellite loci FCA045, FCA077, FCA008, and FCA096 are highly variable molecular markers which were used to determine the genetic diversity in 148 captive Leopardus sp. The PCR-amplified products of microsatellite loci were characterized in ABI Prism 310 Genetic Analyzer. Allele numbers, heterozygosity, polymorphism information content, exclusive allele number, and shared alleles were calculated. Sixty-five alleles were found and their sizes ranged from 116 to 216 bp in four microsatellite loci. The heterozygosity ranged from 0.36 to 0.81 in Leopardus pardalis, 0.57 to 0.67 in L. tigrinus and 0.80 to 0.92 in L. wiedii. The polymorphism information content was from 0.80 to 0.88 in L. pardalis, 0.76 to 0.88 in L. tigrinus and 0.77 to 0.90 in L. wiedii. The margay (L. wiedii) showed the highest index of polymorphism among the three species in this study. These results imply that microsatellite DNA markers can help in the study of the genetic diversity of Leopardus specimens.

Genetic diversity of microsatellite loci in Leopardus pardalis, Leopardus wiedii and Leopardus tigrinus

The microsatellite loci FCA045, FCA077, FCA008, and FCA096 are highly variable molecular markers which were used to determine the genetic diversity in 148 captive Leopardus sp. The PCR-amplified products of microsatellite loci were characterized in ABI Prism 310 Genetic Analyzer. Allele numbers, heterozygosity, polymorphism information content, exclusive allele number, and shared alleles were calculated. Sixty-five alleles were found and their sizes ranged from 116 to 216 bp in four microsatellite loci. The heterozygosity ranged from 0.36 to 0.81 in Leopardus pardalis, 0.57 to 0.67 in L. tigrinus and 0.80 to 0.92 in L. wiedii. The polymorphism information content was from 0.80 to 0.88 in L. pardalis, 0.76 to 0.88 in L. tigrinus and 0.77 to 0.90 in L. wiedii. The margay (L. wiedii) showed the highest index of polymorphism among the three species in this study. These results imply that microsatellite DNA markers can help in the study of the genetic diversity of Leopardus specimens.

Effects of exogenous hexacosanoic acid on biochemical myelin composition in weaning and post-weaning rats.

X-linked Adrenoleukodistrophy (ALD) is characterized by an increase of very long chain fatty acids (VLCFA) in particular of hexacosanoic acid (HA), in tissues and fluids. The biochemical abnormality is due to the dysfunction of peroxisomal degradation of VLCFA. To-date it is unclear if the demyelination which characterizes this disease is the direct consequence of HA accumulation. In order to investigate whether the large amounts of exogenous HA could affect myelin synthesis, 500 micrograms of this fatty acid dissolved in peanut oil were administered daily and by gavage to newborn rats. Since myelin is actively synthesized during early neonatal life and it can be altered by environmental factors including diet, we analyzed lipid and protein composition of myelin after 20, 30 and 60 days of HA administration. Our results show that exogenous HA is incorporated in myelin where it determines biochemical alterations in normal rats having a functioning peroxisomal system. Even though the differences between controls and treated rats are slight, we observed in test rats, a decrease of 2'3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) activity and of myelin basic protein (MBP) content at any time studied. The decrease of glycolipids (GL) was present only after 20 days of treatment. Since these parameters are related to myelin development, our data lead us to think that the myelin of the treated animals is less mature than that of controls.

[Hypoplasia of the renal artery as a cause of renovascular hypertension. A case report]. / L'ipoplasia dell'arteria renale come causa di ipertensione reno-vascolare. Descrizione di un caso clinico.

The authors report the case of an 8-year-old Libyan boy who presented with hypertension, episodes of vomiting and headaches during the past year. Routine blood tests, including nitrogen and urea clearances, were normal; an ultrasound scan and a urography demonstrated a left kidney smaller than the right. Further tests were carried out to evaluate the morphology and the function of the kidney; a scintigraphy confirmed the hypoplasia of the left kidney with reduction of the glomerular filtration rate. The arteriography of the abdominal aorta was decisive, confirming not only the hypoplasia of the left kidney, but also the presence of a hypoplasic renal artery. The latter appeared to be uniformly hypoplasic, with no signs of segmentary stenosis. This fact suggested the hypothesis of renovascular hypertension caused by a hypoplasia of the renal artery. The medical treatment with enalapril gave good results, with normalization of the pressure levels after 5 days of administration. However, only nephrectomy is able to give excellent long-term results.

Traumatic trochanteric bursitis.

The trochanteric bursa is anatomically quite susceptible to traumatic injury. However, specific reports of traumatic trochanteric bursitis in the literature are lacking. This may be due to failure to recognize this condition and to differentiate it from other pain syndromes involving the hip. This case report demonstrates the clinical presentation and management of traumatic trochanteric bursitis.

[Intestinal infections in the course of continuous elemental enteral feeding by way of the jejunum in the infant]. / Infezioni intestinali in corso di alimentazione enterale elementare continua per via digiunale nel lattante.

The authors take an interest in the study of the intestinal infections during continuous enteral elemental feeding by jejunum in infants. They discuss the pathogenesis and indicate the measures in order to prevent that complications. The Authors report 9 cases of infants on whom an enteral (jejunal) elemental feeding was made up: on the basis of their experiences they consider that the occurrence of intestinal infections does not compromise the employing of the enteral feeding in front of other methods of artificial nutrition.