Transoperative refusion: a simple and safe method in emergency surgery

The objective is to reinforce the importance of blood reinfusion as a cheap, safe and simple method, which can be used in small hospitals, especially those in which there is no blood bank. Moreover, even with the use of devices that perform the collection and filtration of blood, more recent studies show that the cost-benefit ratio is much better when autologous transfusion is compared with blood transfusions, even when there is injury to hollow viscera and blood contamination. It is known that the allogeneic blood transfusion carries a number of risks to patients, among them are the coagulation disorders mediated by excess enzymes in the conserved blood, and deficiency in clotting factors, mainly the Factor V, the proacelerin. Another factor would be the risk of contamination with still unknown pathogens or that are not investigated during screening for selection of donors, such as the West Nile Fever and Creutzfeldt-Jacob, better known as "Mad Cow" disease. Comparing both methods, we conclude that blood autotransfusion has numerous advantages over heterologous transfusion, even in large hospitals. We are not against blood transfusions, just do not agree that the patient's own blood is discarded without making sure there will be enough blood in stock to get him out of the hemorrhagic shock.

Screening of antigenemia and isolation of Cryptococcus neoformans and C. gattii from cloaca and crop of birds in the state of Paraná, Brazil

Cryptococcus neoformans and C. gattii are associated with dry bird excreta but rarely recovered from birds' digestive tract. The objective of the present study was (1) to verify the existence of C. neoformans and C. gattii in crop and cloaca of wildlife and captivity birds hypothesizing about a possible primary source of this yeast in the excreta, and (2) to determine the fungi's invasive capability in avian species through latex agglutination. For that purpose, 172 cloacal and 77 crop samples of domestic pigeon, Passerine, and Psittacine birds were collected. None of these samples was positive, suggesting that the yeast is not saprobiotic in the digestive tract of these birds. Only one out of 82 serum samples collected from pigeons and Psittacine birds was positive (title 1:2) showing that Cryptococcus sp. probably has a low invasive capability in birds, and is thus considered only a dry excreta colonizer.

Luminescent Lanthanide Reporters: New Concepts for Use in Bioanalytical Applications

Lanthanides represent the chemical elements from lanthanum to lutetium. They intrinsically exhibit some very exciting photophysical properties, which can be further enhanced by incorporating the lanthanide ion into organic or inorganic sensitizing structures. A very popular approach is to conjugate the lanthanide ion to an organic chromophore structure forming lanthanide chelates. Another approach, which has quickly gained interest, is to incorporate the lanthanide ions into nanoparticle structures, thus attaining improved specific activity and binding capacity. The lanthanide-based reporters usually express strong luminescence emission, multiple narrow emission lines covering a wide wavelength range, and exceptionally long excited state lifetimes enabling timeresolved detection. Because of these properties, the lanthanide-based reporters have found widespread applications in various fields of life. This study focuses on the field of bioanalytical applications. The aim of the study was to demonstrate the utility of different lanthanide-based reporters in homogeneous Förster resonance energy transfer (FRET)-based bioaffinity assays. Several different model assays were constructed. One was a competitive bioaffinity assay that utilized energy transfer from lanthanide chelate donors to fluorescent protein acceptors. In addition to the conventional FRET phenomenon, a recently discovered non-overlapping FRET (nFRET) phenomenon was demonstrated for the first time for fluorescent proteins. The lack of spectral overlap in the nFRET mechanism provides sensitivity and versatility to energy transfer-based assays. The distance and temperature dependence of these phenomena were further studied in a DNA-hybridization assay. The distance dependence of nFRET deviated from that of FRET, and unlike FRET, nFRET demonstrated clear temperature dependence. Based on these results, a possible excitation mechanism operating in nFRET was proposed. In the study, two enzyme activity assays for caspase-3 were also constructed. One of these was a fluorescence quenching-based enzyme activity assay that utilized novel inorganic particulate reporters called upconverting phosphors (UCPs) as donors. The use of UCPs enabled the construction of a simple, rather inexpensive, and easily automated assay format that had a high throughput rate. The other enzyme activity assay took advantage of another novel reporter class, the lanthanidebinding peptides (LBPs). In this assay, energy was transferred from a LBP to a green fluorescent protein (GFP). Using the LBPs it was possible to avoid the rather laborious, often poorly repeatable, and randomly positioned chemical labeling. In most of the constructed assays, time-resolved detection was used to eliminate the interfering background signal caused by autofluorescence. The improved signal-to-background ratios resulted in increased assay sensitivity, often unobtainable in homogeneous assay formats using conventional organic fluorophores. The anti-Stokes luminescence of the UCPs, however, enabled the elimination of autofluorescence even without time-gating, thus simplifying the instrument setup. Together, the studied reporters and assay formats pave the way for increasingly sensitive, simple, and easily automated bioanalytical applications.

Paracoccidioides brasiliensis infection in dogs from Western Brazilian Amazon

The objective of the study was to evaluate Paracoccidioides brasiliensis infection in urban dogs from the municipality of Monte Negro, Rondonia, Western Brazilian Amazon. The serum samples (n=126) were analyzed by indirect ELISA and the immunodiffusion test using P. brasiliensis gp43 and exoantigen as antigens, respectively. A positivity of 54.8% was observed only in the ELISA test and no statistical difference was observed in the seroprevalence in relation to age or sex. This is the first paracoccidioidomycosis survey carried out with dogs from the Western Brazilian Amazon. The higher positivity rates of P. brasiliensis infection observed in this study suggest that veterinarians must be alert to detect new cases of natural disease in dogs living in paracoccidioidomycosis endemic areas.

Feasibility of ASH DEC‐ process in treating sewage sludge and manure ash in Finland

In Finland the thermal treatment of sewage sludge has been moderate in 21th century. The reason has been the high moisture content of sludge. During 2005-2008, 97-99% of sewage sludge was utilized in landscaping and agriculture. However agricultural use has been during 2005-2007 less than 3 %. The aim of national waste management plan is that by 2016 100% of sludge is used either as soil amendment or energy. The most popular utilization method for manure is spreading it on arable land. The dry manures such as poultry manure and horse manure could also be used in incineration. The ashes could be used as fertilizers and while it is not suitable as a starter fertilizer, it is suitable in maintaining P levels in the soil. One of the main drivers for more efficient nutrient management is the eutrophication in lakes and the Baltic See. ASH DEC process can be used in concentrating phosphorus rich ashes while separating the heavy metals that could be included. ASH DEC process uses thermochemical treatment to produce renewable phosphate for fertilizer production. The process includes mixing of ashes and chlorine donors and subsequent treatment in rotary kiln for 20 min in temperature of 900 – 1 050 oC. The heavy metals evaporate and P-rich product is obtained. The toxic substances are retained in air pollution control system in form of mixed metal hydroxides. The aim of conducting this study is to estimate the potential of ASH DEC process in treating phosphorus rich ashes in Finland. The masses considered in are sewage sludge, dry manure from horses, and poultry and liquid pig manure. To date the usual treatment method for sewage sludge in Finland is composting or anaerobic digestion. Part of the amount of produced sewage sludge (800 kt/a fresh mass and 160 kt/a TS) could also be incinerated and the residual ashes used in ASH DEC process. Incinerating only manure can be economically difficult to manage because the incineration of manure is in Finland considered as waste incineration. Getting a permit for waste incineration is difficult and also small scale waste incineration is too expensive. The manure could act as an additional feedstock in counties with high density of animal husbandry where the land area might not be enough for spreading of manure. Now when the manure acts as a supplementary feedstock beside sludge, the ash can’t be used directly as fertilizer. Then it could be used in ASH DEC process. The perquisite is that the manure producers could pay for the incineration, which might prove problematic.

Insights on bovine genetic engineering and cloning

Transgenic technology has become an essential tool for the development of animal biotechnologies, and animal cloning through somatic cell nuclear transfer (SCNT) enabled the generation of genetically modified animals utilizing previously modified and selected cell lineages as nuclei donors, assuring therefore the generation of homogeneous herds expressing the desired modification. The present study aimed to discuss the use of SCNT as an important methodology for the production of transgenic herds, and also some recent insights on genetic modification of nuclei donors and possible effects of gene induction of pluripotency on SCNT.

Reproductive efficiency of asymptomatic Theileria equi carriers mares submitted to an embryo transfer program

This study aimed to assess and evaluate the effects of Theileria equi infection on embryonic recovery, gestation and early embryonic loss. Thirteen Mangalarga Marchador Theileria equi positive donors (diagnosed through nested-PCR) and 40 embryos receptors were used. Donors were submitted to two embryo collections in two consecutive estrous cycles (GId); after, the same mares were treated with imidocarb dipropionate (1.2mg/kg IM.) in order to collect more embryos in two more estrous cycles (GIId). Receptors were divided into two groups (control and with treated) with 20 animals each, where one group was the control (GIr) and the other one (GIIr) treated with 1.2mg/kg IM of imidocarb dipropionate assessing the gestation rate at 15, 30, 45 and 60 days. After 52 embryo collections, the embryonic recovery rates were 53.84% (14/26) and 65.38% (17/26) (p> 0.05) for GId and GIId, respectively. The gestation rate was 70% (14/20) (p>0.05) at 15, 30, 45 and 60 days in group GIr and for GIIr was 85% (17/20) (p>0.05) at 15 days, 80% (16/20) (p>0.05) at 30, 45 and 60 days. The treatment with imidocarb dipropionate did not cause significant improvement in the reproductive efficiency at an ET program.

Controle do arroz vermelho através do herbicida sulfosate isolado e em misturacom adjuvantes

O presente trabalho teve como objetivo avaliar a eficiência do herbicida sulfosate, aplicado isoladamente e em misturas com diferentes adjuvantes, comparado com o herbicida glifosate, ambos aplicados com baixo volume de calda, no controle da planta daninha arroz vermelho (Oryza sativa L.) sob aplicação em condições de préplantio da cultura do arroz. O experimento foi conduzido em campo, sendo a área experimental instalada na cultura de arroz irrigado, variedade IAC-101. A área de pousio recebeu uma lâmina de água de 5 a 10cm de pro fundidade por dois dias, e após a retirada, o arroz vermelho começou a germinar. Quando o mesmo atingiu cerca de 20-25cm de altura, procedeu-se a aplicação de herbicidas de manejo (pré-plantio) . Após 16 dias da aplicação foi efetuado o plantio da cultura com densidade de 60-80 sementes/metro linear e profundidade de 3cm, sendo o espaçamento utilizado de 21,8cm entre linhas. Foram testados tratamentos com os seguintes produtos: sulfosate a 1,20; 1,44 e 1,68Kg i.a./ha e glifosate a 1,20; 1,44 e 1,68Kg i.a./ha, ambos os produtos aplicados isolados e em misturas com os seguintes adjuvantes: Poliglicol, Organosilicone e Amina Graxa, além de uma testemunha não capinada. Para a avaliação do efeito dos tratamentos empregados foram realizadas as seguintes avaliações: fitotoxicidade aperente, porcentagem de controle do arroz vermelho; altura do arroz vermelho (em cm); estande da cultura (no de per filhos/metro linear) e altura da cultura (em cm). Através dos resultados obtidos pode-se concluir que os herbicidas sulfosate, glifosate nas formulações comerciais Rodeo e Roundup, nas doses de 1,20; 1,44 e 1,68Kg i.a./ha, aplicados isolados (exceto o Rodeo) e conjuntamente com os adjuvantes poliglicol (Mojante), organosilicone (Silwet) e amina graxa (Frigate) (exceto Roundup, que foi utilizado apenas com o adjuvante amina graxa e sozinho), propiciaram excelente nível de controle do arroz vermelho, quando aplicados em pós-emergência e área total, e pré-plantio da cultura de arroz, implantada no sistema de plantio direto. Também foi verificado que não ocorreu diferença quanto à eficiência entre sulfosate e glifosate no controle do arroz vermelho, ambos controlando de forma eficiente a planta daninha.

Avaliação de tratamentos químicos e mecânicos no controle de plantas daninhas na cultura do algodão

O presente trabalho teve como objetivo avaliar a eficiência de diferentes combinações de tratamentos herbicidas integrado, aplicados em PPI, pré, pós, cultivo mecânico e jato dirigido em cultura de algodão (Gossypium hirsutum L.). O experimento foi conduzido em campo, sendo a área experimental instalada na cultura de algodão cultivar IAC-20. Foram testados tratamentos com os seguintes produtos: trifluralina na dose de 1,068 kg i.a./ha; alachlor na dose de 1,584 kg i.a./ha; M.S.M.A. nas doses de 1,44 e 1,68 kg i.a./ha; nicosulfuron nas doses de 0,0201; 0,030 e 0,040 kg i.a./ha; diuron nas doses de 1,0 e 1,5 kg i.a./ha; lactofen na dose de 0,18 kg i.a./ha; cyanazine nas doses de 1,0 e 1,5 kg i.a./ha e diuron nas doses de 1,0 e 1,5 kg i.a./ha.Os tratamentos herbicidas foram efetuados em pré-plantio incorporado (PPI), pré-emergência (PRE), pós-emergência (POS), cultivo mecânico (CM) e jato dirigido (JD). Os resultados obtidos levaram a concluir que o sistema de trifluralina (PPI), alachlor (PRE), CM, M.S.M.A. + lactofen apresentaram a maior produtividade. Os sistemas herbicidas compostos por aplicação de trifluralina (PPI), alachlor (PRE), M.S.M.A. (POS), CM ao redor dos 50 a 60 D.A.P. (dias após o plantio) e a aplicação de jato dirigido de diuron, lactofen + cyanazine, lactofen + diuron, M.S.M.A. + cyanazine, M.S.M.A. + lactofen e M.S.M.A. + diuron, propiciaram de forma geral controle das principais plantas daninhas.

Influência do manejo da palhada de capim-braquiária (Brachiaria decumbens) sobre o desenvolvimento inicial de soja (Glycine max) e amendoim-bravo (Euphorbia heterophylla)

Com o objetivo de avaliar os efeitos de diferentes manejos de palhada de capim-braquiária sobre o desenvolvimento inicial da cultura de soja e da planta daninha amendoimbravo, foi conduzido um experimento em condições de casa de vegetação no NuPAMFCA/UNESP, BotucatuSP. Os tratamentos utilizados foram: manejo da palhada na superfície do solo + irrigação superficial (T1); manejo da palhada na superfície do solo + irrigação subsuperficial (T2); palhada incorporada ao solo (T3); e testemunha sem cobertura (T4). A palhada foi colhida no campo 30 dias após dessecação com o herbicida glyphosate (1,44 g i.a. ha-1). O delineamento experimental utilizado foi o inteiramente casualizado, com sete repetições, sendo as unidades experimentais vasos plásticos, com a soja e o amendoim-bravo semeados paralelamente, em linhas distintas. O T1 reduziu significativamente o índice de velocidade de germinação (IVG) e a altura das plântulas de soja aos 5 e 10 dias após a emergência (DAE), ao contrário do amendoim-bravo, o qual não sofreu interferência dos tratamentos estudados, constituindo-se em uma planta-problema para sistemas produtivos com palhada de capim-braquiária. Os resultados da análise de crescimento (TCA - taxa de crescimento absoluto, TCR - taxa de crescimento relativo e TAL - taxa de assimilação líquida) das plântulas de soja e amendoim-bravo apresentaram valores máximos aos 15 DAE, com exceção do T3 para soja, o qual reduziu expressivamente o desenvolvimento em relação aos demais tratamentos.

Spotlight: A Blacklight Plugin for Showcasing Digital Collections

Presentation at Open Repositories 2014, Helsinki, Finland, June 9-13, 2014

Inheritance of blast resistance in rice to two Pyricularia grisea races, IB-1 and IB-9

Seven sources of resistance to the two predominant races IB-1 and IB-9 of the rice blast pathogen Pyricularia grisea were selected based on leaf blast reaction in tests conducted under controlled greenhouse conditions. Crosses involving resistant and susceptible parents were made to study the inheritance of the disease reaction for different sources of resistance. The F1 and F2 progenies of all crosses, including backcrosses to resistant and susceptible parents, were tested for reaction to leaf blast. The data showed that resistance is controlled by one to three genes that segregate independently in most of the donors. Non-allelic interaction among resistance genes, including dominant epistasis, was identified.

Role of nitric oxide and superoxide in Giardia lamblia killing

Giardia lamblia trophozoites were incubated for 2 h with activated murine macrophages, nitric oxide (NO) donors or a superoxide anion generator (20 mU/ml xanthine oxidase plus 1 mM xanthine). Activated macrophages were cytotoxic to Giardia trophozoites (~60% dead trophozoites). This effect was inhibited (>90%) by an NO synthase inhibitor (200 µM) and unaffected by superoxide dismutase (SOD, 300 U/ml). Giardia trophozoites were killed by the NO donors, S-nitroso-acetyl-penicillamine (SNAP) and sodium nitroprusside (SNP) in a dose-dependent manner (LD50 300 and 50 µM, respectively). A dual NO-superoxide anion donor, 3-morpholino-sydnonimine hydrochloride (SIN-1), did not have a killing effect in concentrations up to 1 mM. However, when SOD (300 U/ml) was added simultaneously with SIN-1 to Giardia, a significant trophozoite-killing effect was observed (~35% dead trophozoites at 1 mM). The mixture of SNAP or SNP with superoxide anion, which yields peroxynitrite, abolished the trophozoite killing induced by NO donors. Authentic peroxynitrite only killed trophozoites at very high concentrations (3 mM). These results indicate that NO accounts for Giardia trophozoite killing and this effect is not mediated by peroxynitrite

The myth of nitric oxide in central cardiovascular control by the nucleus tractus solitarii

Considerable evidence suggests that nitroxidergic mechanisms in the nucleus tractus solitarii (NTS) participate in cardiovascular reflex control. Much of that evidence, being based on responses to nitric oxide precursors or inhibitors of nitric oxide synthesis, has been indirect and circumstantial. We sought to directly determine cardiovascular responses to nitric oxide donors microinjected into the NTS and to determine if traditional receptor mechanisms might account for responses to certain of these donors in the central nervous system. Anesthetized adult Sprague Dawley rats that were instrumented for recording arterial pressure and heart rate were used in the physiological studies. Microinjection of nitric oxide itself into the NTS did not produce any cardiovascular responses and injection of sodium nitroprusside elicited minimal depressor responses. The S-nitrosothiols, S-nitrosoglutathione (GSNO), S-nitrosoacetylpenicillamine (SNAP), and S-nitroso-D-cysteine (D-SNC) produced no significant cardiovascular responses while injection of S-nitroso-L-cysteine (L-SNC) elicited brisk, dose-dependent depressor and bradycardic responses. In contrast, injection of glyceryl trinitrate elicited minimal pressor responses without associated changes in heart rate. It is unlikely that the responses to L-SNC were dependent on release of nitric oxide in that 1) the responses were not affected by injection of oxyhemoglobin or an inhibitor of nitric oxide synthesis prior to injection of L-SNC and 2) L- and D-SNC released identical amounts of nitric oxide when exposed to brain tissue homogenates. Although GSNO did not independently affect blood pressure, its injection attenuated responses to subsequent injection of L-SNC. Furthermore, radioligand binding studies suggested that in rat brain synaptosomes there is a saturable binding site for GSNO that is displaced from that site by L-SNC. The studies suggest that S-nitrosocysteine, not nitric oxide, may be an interneuronal messenger for cardiovascular neurons in the NTS

Correlation of discocyte frequency and ATP concentration in preserved blood: A morphological indicator of red blood cell viability

Red blood cells (RBC) are viable if kept in an adequate preservative solution, although gradual changes in morphology and metabolism may occur. There is a gradual decrease in adenosine-5'-triphosphate (ATP) concentration, pH, glucose consumption, and enzyme activity during preservation. The normal discocyte shapes are initially replaced by echinocytes and stomatocytes and, at final stages, by spherocytes, the last step before splenic sequestration. Post-transfusional survival has been correlated with the ATP concentration. RBC preserved in ADSOL, a solution containing adenine, dextrose, sodium chloride, and mannitol, are viable for transfusion for up to 6 weeks. Erythrocytes from 10 blood units taken from healthy adult donors were preserved for 12 weeks in ADSOL at 4oC. We now report a significant correlation (r2 = 0.98) between the percentage of discocytes (89 to 7%) and ATP (100 to 10%) concentration in ADSOL-preserved RBC. The results suggest that the percent of discocyte shapes used as an indicator of ATP concentration may be a useful indicator for quality control of RBC viability in centers which have limited assay facilities.