CCS e amiloide A do leite de quartos mamários tratados na secagem com plasma rico em plaquetas autólogo, associado ou não a antibiótico / [CCS and Amyloid A of milk from breast quarters treated in drying with autologous platelet-rich plasma, with or without antibiotics]

Este estudo teve como objetivo avaliar biomarcadores de mastite após terapia da secagem com plasma rico em plaquetas (PRP), associado ou não a antibiótico. Trinta e seis quartos mamários foram utilizados em três tratamentos (T): T1 - antibiótico (ATB), T2 - PRP e T3 - PRP + ATB. Amostras de leite foram coletadas antes da secagem (A1), no parto (D0) e 14, 30 e 60 dias pós-parto (D14, D30 e D60), para determinar contagem de células somáticas (CCS) e amiloide A (AA). O delineamento foi inteiramente ao acaso, com arranjo em parcelas subdivididas (SigmaPlot®). Dados de CCS foram transformados (log 10). As médias foram comparadas utilizando-se testes de Tukey ou Holm-Sidak (P<0,05). A CCS em A1 foi elevada em todos os grupos (P>0,05). No D30, CCS foi maior em T2 (P<0,05), igualando-se no D60. Não houve diferença na AA entre Ts em qualquer dia de coleta (P>0,05). Houve diferença nos momentos de coleta (P<0,05), A1 maior que D14 e D30. Houve uma correlação positiva fraca com CCS (0,280). Os tratamentos foram semelhantes em manter a saúde da glândula mamária na lactação subsequente. O PRP intramamário pode ser usado para terapia de vaca seca em casos de mastite subclínica.(AU)

This study aimed to evaluate biomarkers of mastitis after autologous platelet rich plasma (PRP) dry cow therapy, associated or not with antibiotic, compared to conventional treatment. Thirty-six mammary quarters were used in three treatments (T): T1 - antibiotic, T2 - PRP and T3 - PRP + ATB. Milk samples were collected before drying (A1), on calving (D0) and at 14, 30 and 60 days postpartum (D14, D30 and D60), for Somatic Cell Count (CCS) and amyloid dosage A (AA). The design was completely randomized with arrangement in subdivided plots (SigmaPlot ®). Data from CCS were transformed in log10. Means were compared using the Tukey or Holm-Sidak tests, at a 95% confidence level (P<0.05). CCS in A1 was elevated in allgroups (P>0.05). On D30 CCS was higher in T2 (P<0.05), matching D60. There was no difference in AA among T on any day of collection (P>0.05). There was difference in collection moments (P<0.05), A1 different from D14 and D30. There was a weak positive correlation with CCS (0.280). Three treatments were similar in maintaining the health of the mammary gland at subsequent lactation. PRP intramammary can be used for dry cow therapy in subclinical mastitis.(AU)

Novel PCR primers for improved detection of Mycobacterium leprae and diagnosis of leprosy.

AIMS: Diagnosis of leprosy, a chronic infection caused by Mycobacterium leprae, predominantly depends on clinical manifestations and histopathological analysis, hampering rapid and accurate diagnostics. Our aim was to increase accuracy of leprosy diagnosis by improving M. leprae's DNA detection based on polymerase chain reaction (PCR) technique using new specific primers for the RLEP repetitive sequence. METHODS AND RESULTS: The specific target region, RLEP, of M. leprae's genome was selected based on comparative genomics. After confirming the specificity of this region, using blastn analysis, primers were designed and tested for their in silico specificity. To evaluate the specificity and sensitivity of these primers in vitro, 184 blood samples from patients were used in qPCR. The new primer pair LYON1/LYON2 produced 91% positive samples, whereas the current primer pair LP1/LP2 produced 46%. Specificity and DNA detection limit test were carried out to compare the efficiency of the developed primer pair. The LYON1/LYON2 primer showed 100% specificity, whereas LP1/LP2 showed 64%. The DNA detection limit of LYON1/LYON2 was 10 copies of bacterial genomes per millilitre, whereas LP1/LP2 was 1000 copies of bacterial genomes per millilitre. CONCLUSIONS: In conclusion, the developed LYON1/LYON2 primer pair presented to be a specific and sensitive new molecular marker for the diagnosis of leprosy. SIGNIFICANCE AND IMPACT OF THE STUDY: The development of a specific primer pair for the detection of the M. leprae genome through qPCR technique contributes to a fast, sensitive and specific diagnosis, which is essential to prevent spreading and progression of this disease.

Systemic absortion and adverse effects of topical ocular use of ketorolac tromethamine and sodium diclofenac in New Zealand rabbits for 90 days / Absorção sistêmica e efeitos adversos do uso tópico ocular por 90 dias de cetorolaco de trometamina e diclofenacode sódio em coelhos da raça Nova Zelândia

The effect of the systemic absorption of 0.1% diclofenac sodium (DS) eyedrop was compared to that of 0.5% ketorolac tromethamine (KT) in female New Zealand white rabbits treated on both eyes three times a day for 90 days. The rabbits were divided in three groups of six animals (n= 18): KT group, DS group, and control (Co) group, in which saline (0.9% NaCl) solution was instilled. Water and food consumption were measured daily, clinical examination was performed weekly, and blood samples were collected every 30 days for laboratory examination. The plasma was analyzed for the presence of KT and DS by solid-phase extraction (SPE) associated with mass spectrometry (MS). Systemic absorption of these drugs was confirmed by SPE-MS, allowing their separation and identification in the plasma. At the end of the treatment, the animals were euthanized and necropsied, and no macroscopic or microscopic changes were found. This observation confirmed the laboratory results, which were within normal reference standards for the species. According to the results obtained, it can be concluded that treatment with eyedrops containing KT and DS for 90 days in healthy rabbits does not cause adverse systemic effects.(AU)

Comparou-se o efeito da absorção sistêmica do colírio de diclofenaco de sódio 0,1% (DS) em relação ao de cetorolaco de trometamina 0,5% (CT) em coelhas da raça Nova Zelândia, tratadas nos dois olhos, três vezes ao dia, por 90 dias. As coelhas foram separadas em três grupos de seis animais (n=18): grupo CT, grupo DS e grupo controle (Co), no qual foi instilada solução fisiológica (NaCl 0,9%). Os consumos de água e ração foram mensurados diariamente, os exames clínicos foram realizados semanalmente e o sangue foi coletado a cada 30 dias para realização de exames laboratoriais. O plasma foi analisado para detectar a presença de CT e DS por extração em fase sólida (SPE) associada à espectrometria de massas (MS). A absorção sistêmica desses fármacos foi confirmada por SPE-MS, permitindo sua separação e identificação no plasma. Ao final do tratamento, os animais foram eutanasiados e necropsiados, e não foram encontradas alterações macroscópicas ou microscópicas. Essa observação confirmou os resultados laboratoriais, que estavam dentro dos padrões de referência para a espécie. De acordo com os resultados obtidos, pode-se concluir que o tratamento com colírio contendo KT e DS, por 90 dias, em coelhos saudáveis, não causa efeitos adversos sistêmicos.(AU)

How do geometry-related parameters influence the clinical performance of orthodontic mini-implants? A systematic review and meta-analysis.

The aim of this systematic review and meta-analysis was to investigate how parameters related to geometry influence the clinical performance of orthodontic mini-implants (MIs). Systematic searches were performed in electronic databases including MEDLINE, Scopus, Web of Science, Virtual Health Library, and Cochrane Library and reference lists up to March 2016. Eligibility criteria comprised clinical studies involving patients who received MIs for orthodontic anchorage, with data for categories of MI dimension, shape, and thread design and insertion site, and evaluated by assessment of primary and secondary stability. Study selection, data extraction, quality assessment, and a meta-analysis were carried out. Twenty-seven studies were included in the qualitative synthesis: five randomized, eight prospective, and 14 retrospective clinical studies. One study with a serious risk of bias was later excluded. Medium and short MIs (1.4-1.9mm diameter and 5-8mm length) presented the highest success rates (0.87, 95% CI 0.80-0.92). A maximum insertion torque of 13.28Ncm (standard error 0.34) was observed for tapered self-drilling MIs in the mandible, whereas cylindrical MIs in the maxilla presented a maximum removal torque of 10.01Ncm (standard error 0.17). Moderate evidence indicates that the clinical performance of MIs is influenced by implant geometry parameters and is also related to properties of the insertion site. However, further research is necessary to support these associations.

Progenitor cells and TNF-alpha involvement during morphological changes in pancreatic islets of obese mice.

Overnutrition during pregnancy and lactation lend increasing support to the development of obesity and several chronic diseases in adulthood such as type 2 diabetes mellitus, which leads to beta-cell dysfunction and insulin resistance. In this work, we aimed to study the effects of early life overnutrition on the development of obesity, analyzing the morphological changes, expression of TNF-α, and also the stem cell marker CD133 in the pancreatic islets of young and adult mice. Overnutrition during lactation phase was used as an experimental model to induce obesity. The animals were analyzed at 28 and 150 days of age, when pancreata were collected for histological, ultrastructural and western blotting analysis. The results showed that islet hypertrophy is established in obese groups at day 28 and remained until adulthood. CD133+ cells were observed as small cells within pancreatic islets in both control and obese young mice. However, at day 150, these cells were observed only in the islet peripheries and near ducts of the obese group. Furthermore, TNF-α expression in pancreatic islets was increased in both young and adult obese groups when compared to control groups. This work shows interesting data about CD133 receptor and TNF-α roles in the pancreas during obesity development.

Determination of the relevant magnetic interactions in low-dimensional molecular materials: the fundamental role of single crystal high frequency EPR.

A new one-dimensional copper(II) complex with formula [Cu(hfac)(2)(N(3)TEMPO)](n) (hfac = hexafluoroacetylacetonate and N(3)TEMPO = 4-azido-2,2,6,6-tetramethylpiperidine-1-oxyl) has been synthesized and investigated by X-ray crystallography, magnetometry and multifrequency single crystal EPR. The system crystallizes in the P1 space group with two non equivalent copper(II) ions in the unit cell, the two nitroxide radicals being coordinated to Cu(1) in axial positions. The copper(II) ions are bridged by N(3)TEMPO radicals resulting in a zig-zag chain structure. The magnetic susceptibility data were at first satisfactorily modeled assuming an alternating spin chain along the monodimensional covalent skeleton, with a ferromagnetic interaction between Cu(1) and the nitroxide moieties and a weaker antiferromagnetic interaction between these and Cu(2) (J(1) = -13.8 cm(-1), J(2) = +2.4 cm(-1)). However, single crystal EPR studies performed at the X- and W-band clearly demonstrate that the observed magnetic monodimensional character of the complex is actually due to the intermolecular contacts involving N(3)TEMPO ligands. This prompted us to fit the magnetic data using a consistent model, pointing out the fundamental role of single crystal EPR data in defining a correct model to describe the magnetic properties of molecular low dimensional systems.

Synthesis of 4-acyl-1H-1,2,3-triazolic nucleosides.

Two simple regiospecific methodologies based on triazolic ring construction in the course of synthesis were applied for the synthesis of 1,2,3triazolic nucleoside analogues. The cycloaddition reactions between diazomalonaldehyde and appropriate glycosylamine derivatives were rather effective, producing the desired nucleosides 11, 17 and 24. Diazotization of enamines 21a and 21b led to the corresponding triazolic ribonucleoside derivatives 22a and 22b, in good yields. Deprotection reaction of 22a, 22b and 24 was easily achieved by Lewis acid catalysis, producing the corresponding ribonucleosides 23a, 23b and 25.

Ontogenic variations in n-alkanes during somatic embryogenesis of flax (Linum usitatissimum L.).

Hypocotyl segments (HS) of flax seedlings germinated in vitro, were used to induce indirect somatic embryogenesis on solid medium. The composition and distribution of n-alkanes in flax tissues collected at different developmental stages were studied by capillary gas chromatography (GC) and capillary gas chromatography-mass spectrometry (GC-MS). During induction and development of callus from hypocotyl tissues a decrease in the percentage of total lipids was observed. In all types of tissue sampled - HS used as primary explants, HS with differentiating calli at the cut ends (HSC), embryogenic (EC) and non-embryogenic calli (NEC) and somatic embryos (SE) - a skewed-normal distribution of n-alkanes with a low mass range (C(13)-C(21)) were found. The highest content of n-alkanes occurred in the primary hypocotyl explants and in the early stages of callus development. Longer carbon chain n-alkanes were observed only in the mature or differentiated tissues of hypocotyls and SE. Although the n-alkane contents decreased with time, in SE and calli, a significantly lower n-alkane content was observed in EC when compared to NEC independent of the time in culture. These results suggest the utilisation of n-alkanes for heterotrophic cellular growth as well as its mobilisation from EC to developing SE.

Intrauterine device and maternal copper metabolism during lactation.

The effects of intrauterine device (IUD) on maternal copper (Cu) metabolism during breastfeeding was studied in 95 volunteer mothers who chose to use non-hormonal contraceptive methods. They were divided into two groups that were inserted with the IUD-Cu 380A (n = 33), IUD-Cu 200B (n = 29), and a third group that did not use any IUDs served as control (n = 33). Endometrial biopsies, blood, and milk samples were collected before (at 10 weeks postpartum) and 6 weeks after insertion of device for the determination of metabolites associated with copper metabolism, namely, serum ceruloplasmin, and copper concentrations in breast milk and endometrium. Endometrial copper concentration increased in women using IUDs, but was statistically significant (p = 0.001) only in the IUD-Cu 380A group. The increase in endometrial copper concentration did not affect serum ceruloplasmin or milk copper concentrations.

Prevalence of Helicobacter pylori infection in adults submitted to endoscopy in southern Brazil

The most important characteristics of Helicobacter pylori infection were studied in 96 patients submitted endoscopic examination at the University Hospital of the Federal Unlversity of Santa Catarina, Florianópolis southern Brazil. As diagnostic method of the infection we used the presence of Helicobacter pylori on histologic examination, stained by hematoxilin-eosin and Giemsa stains ("gold standard"), immunologic test (ELISA Roche), bacteriologic slide (Gram stained), and urease biopsy test. The rnedian age of the series was 44.6 years (range 14 to 84 years, SD = 18.1), with 56 (58.3 per cent) males. The prevalence of the infection was 78.1 per cent 75/96 and Helicobacter pylori infection was associated with gastritis in 96.6 per cent of the cases (68/75, p < O.0 1). Sixteen of 17 patients with peptic ulcer disease tested positive for the presence of Helicobacter pylori. The infection proportionally more prevalent in the older groups, and there was no statistical gender difference.

[Veno-occlusive disease of the liver as a treatment complication in children with Wilm's tumor]. / Veno-occlusive disease der Leber als Behandlungskomplikation bei Kindern mit Wilmstumor.

We report on three children aged 1 1/2, 2 and 9 1/2 years with Wilms' tumor, who developed a tender hepatomegaly and ascites associated with elevated liver enzymes, anemia and thrombocytopenia during chemotherapy. This clinical picture and liver sonography abnormality are best explained by veno-occlusive disease (VOD) of the liver, while other causes of liver disease could not be identified. Actinomycin D dosage was 0.045 mg/kg as bolus injection in two patients and 0.075 mg/kg split over five days in a third patient. Presumable, this drug was the causative agent. VOD was observed after preoperative and postoperative chemotherapy. No child had received abdominal irradiation. The authors comment on the influence of Actinomycin D as the cause for this unusual liver toxicity.

Doença de Aujeszky: II. Uso do camundongo em teste de eficiência de vacina inativada / Aujeszky disease: II. Efficacy testing of oil-adjuvant vaccine in mice

Realizaram-se vários experimentos, visando determinar a sensibilidade do camundongo de diferentes idades à infecçäo com vírus Aujeszky, vias e dose de inoculaçäo da vacina, bem como verificar o comportamento imunológico à vacina inativada oleosa contra a Doença de Aujeszky. Verificou-se que a via intracerebral é mil vezes mais sensível que a via subcutânea, igualando-se em sensibilidade ao cultivo de células renais de coelho. As vacinas, quando inoculadas por via subcutânea e intraperitoneal, provocaram problemas de reaçäo local e refluxo da vacina, o que implicou em resultados irregulares de proteçäo nas diferentes diluiçöes. As vacinas foram diluídas até 1:32 e a dose vacinal foi de 0,05 ml, no músculo da coxa de camundongos com 10 semanas de idade. Desta forma, foram utilizadas de 1/20 a 1/640 da dose vacinal do suíno (1 ml). As doses de desafio variaram de 3 a 316 DL 50 por cento, via subcutânea. Constatou-se que a Dose Protetora 50 por cento (DP50 por cento) apresentou maior regularidade com a dose de desafio, entre 30 a 100 DL 50 por cento. A DP50 por cento variou entre 11 e 15 nestas doses de desafio, tanto pela vacinaçäo subcutânea como pela intramuscular. Enta última, entretanto, apresentou maior regularidade na proteçäo nas diferentes diluiçöes

Acute respiratory failure after intrathecal methotrexate administration.

This is a report of a case of acute respiratory failure following the administration of intrathecal methotrexate (MTX) for prophylaxis of central nervous system leukemia (CNS) in a 3-year-old girl with acute lymphoblastic leukemia. This could be attributed to acute metabolic or direct toxic effects of methotrexate to the central nervous system tissue. Although the specific agent responsible for this complication following intrathecal injection of MTX is unclear, it is worthwhile to describe this event.

[Silent ischemia in patients with previous myocardial infarct. Correlation of Holter and coronarography]. / Isquémia silenciosa em doentes com prévio enfarte de miocárdio correlação entre Holter e coronariografia.

OBJECTIVE: Our objective was to determine the presence of silent myocardial ischemia and the degree of coronary obstructions. DESIGN: Successively patients with myocardial infarction. SETTING: Patients admitted to the coronary intensive care, later followed in a specialised out-patients consultation. PATIENTS: 62 patients with previous myocardial infarction. INTERVENTIONS: All the patients were submitted to Holter-monitoring and coronary angiography. RESULTS: Silent ischemia was present in 14 (22,6%) of the patients. Seven of these (50%) had 3 vessels disease, 2 (14,3%) had 2 vessels disease, and 5 (35,7%) had 1 vessel disease. Twelve of the patients (86%) had occlusive lesions (greater than 90%), localised in 11 (79%) in the third proximal segment of the artery. CONCLUSIONS: Our results showed that the presence of silent myocardial ischemia was predominantly associated with 3 vessels disease, and with lesions of more than 90% of occlusion, localised in the 1/3 proximal of the vessel. Patients with previous myocardial infarction and silent ischemia may belong to a subgroup with surgical indication. And so, Holter studies in these patients should be followed by angiographic studies for localization of the obstructive lesions.

[Electrolyte changes in acute leukemia in childhood]. / Elektrolytveränderungen bei der akuten Leukämie im Kindesalter.

Electrolyte imbalance in leukemia can be due to either organ infiltration and cell death or to a side effect of cytostatic drugs. From the wide variety of these disturbances seen in acute leukemias in childhood, the excess of potassium is most dangerous. Further electrolyte changes, which are however less evident, are hyperphosphataemia, hyperphosphaturia, and hypocalcaemia. The destruction of a large amount of cells during aggressive induction therapy can boost the electrolyte imbalance and therefore lead to renal failure. Such situations are demonstrated in two cases. Following Vincristine and Cyclophosphamide administration, electrolyte changes such as acute or prolonged decrease of sodium in the serum and urinary loss of sodium are seen frequently. Based on the data from 20 patients with acute lymphoblastic leukemias we describe the dynamics of this process. These changes are probably caused by the syndrome of inadequate ADH-secretion. The clinical importance of these findings are discussed and procedures for improving therapy are set out.