Chemical methods of assessing potentially available organic nitrogen in soil

The need for laboratory methods of obtaining an estimate of the amount of nitrogen (N) likely to be made available for crop growth by mineralization of soil organic matter during the growing season has long been evident, and numerous biological and chemical methods have been proposed. However, the methods showing the most promise are too complicated and time-consuming for use in soil testing laboratories, and there is an urgent need for a simple and rapid chemical method of assessing potentially available organic N in soil;Studies reported led to development of two rapid chemical methods of assessing potentially available organic soil N. One involves determination of the NH(,3)-N produced by steam-distilling the soil sample with pH 11.2 phosphate-borate buffer for 8 min. The other involves determination of the NH(,4)(\u27+)-N produced by heating the soil sample with 2 M KCl at 100(DEGREES)C for 4 hours. Both methods are simple and precise, and their results are not significantly affected by air-drying or air-dry storage of the soil sample before analysis. They are well suited for use in soil testing laboratories because they do not require filtration or transfer steps. Studies using 33 Brazilian soils showed that the results obtained by these methods were highly correlated with those obtained by anaerobic and aerobic incubation methods of assessing potentially available organic N in soil;The two chemical methods developed were further evaluated by applying them to 30 Iowa soils and by comparing their results and those obtained by other chemical methods with the results of the aerobic and anaerobic incubation methods considered to be the best laboratory methods thus far proposed for assessment of potentially available organic N in soil. The chemical methods used included the acid KMnO(,4) method, the alkaline KMnO(,4) method, the CaCl(,2)-autoclave method, and the NaHCO(,3) UV method. The data obtained showed that the results of the two chemical methods developed were highly correlated with the results of the incubation techniques used for comparison and that the correlations observed with these two methods were higher than those observed with the previously proposed chemical methods. It is concluded that these two rapid and simple chemical methods are the best chemical methods thus far developed for assessment of potentially available organic N in soil and that they deserve consideration for use in soil testing laboratories

Examining the potential for porcine-derived islet cells to harbour viral pathogens

With an onus on safety in the potential use of porcine islet cells as a treatment for diabetes, the use of animals lacking exogenous pathogens is clearly important and multilevel screening strategies have been presented on testing animals and the product. In this study, we wished to investigate whether islet cells indeed harboured the same viral pathogens of concern in the source animal. PMBC and islet cells from both adult and neonatal source animals were directly compared and tested for PCMV, PLHV, PCV2, PPV and HEV using both molecular and serological assays. Adult PBMC were found positive for all viruses with the exception of PCV2 and HEV. Neonatal PBMC were only found positive for PCMV and HEV. All animals were found negative for HEV antibodies. Interestingly, islet cells were negative for all viruses tested regardless of status in the animal-derived PBMC. Given that other laboratories have demonstrated the lack of virus detection during the culture of islets, this study also demonstrates that the hygiene status of the herd may not reflect the status of the product. This is important for establishing guidelines for any risk evaluation and mitigation process utilised during product manufacture

Assessment of porcine endogenous retrovirus transmission across an alginate barrier used for the encapsulation of porcine islets

BACKGROUND: Subcutaneous implantation of a macroencapsulated patch containing human allogenic islets has been successfully used to alleviate type 1 diabetes mellitus (T1DM) in a human recipient without the need for immunosuppression. The use of encapsulated porcine islets to treat T1DM has also been reported. Although no evidence of pathogen transfer using this technology has been reported to date, we deemed it appropriate to determine if the encapsulation technology would prevent the release of virus, in particular, the porcine endogenous retrovirus (PERV). METHODS: HEK293 (human epithelial kidney) and swine testis (ST) cells were co-cultured with macroencapsulated pig islets embedded in an alginate patch, macroencapsulated PK15 (swine kidney epithelial) cells embedded in an alginate patch and free PK15 cells. Cells and supernatant were harvested at weekly time points from the cultures for up to 60 days and screened for evidence of PERV release using qRT-PCR to detect PERV RNA and SG-PERT to detect reverse transcriptase (RT). RESULTS: No PERV virus, or evidence of PERV replication, was detected in the culture medium of HEK293 or pig cells cultured with encapsulated porcine islets. Increased PERV activity relative to the background was not detected in ST cells cultured with encapsulated PK15 cells. However, PERV was detected in 1 of the 3 experimental replicates of HEK293 cells cultured with encapsulated PK15 cells. Both HEK293 and ST cells cultured with free PK15 cells showed an increase in RT detection. CONCLUSIONS: With the exception of 1 replicate, there does not appear to be evidence of transmission of replication competent PERV from the encapsulated islet cells or the positive control PK15 cells across the alginate barrier. The detection of PERV would suggest the alginate barrier of this replicate may have become compromised, emphasizing the importance of quality control when producing encapsulated islet patches

Cellular xenotransplantation of animal cells into people: benefits and risk

The main benefit of xenotransplantation is its potential to overcome the worldwide organ shortage experienced in allotransplantation. Allogeneic transplantation is the only successful therapy for several life-threatening diseases, with cell, tissue or organ donation only partially meeting the demand and many patients dying while waiting for treatment. With supply falling short of demand, it is foreseen that the use of porcine material may at some stage overcome the existing gap between organ availability and clinical need. Recently, pig islet cells have been utilised in clinical trials, with safety being demonstrated. Indeed, pig-derived cells present several advantages: i) porcine cells have a stable function and differentiation pattern and are not tumorigenic; ii) pig cells have been shown to meet the physiological needs in large animal models; iii) the source of pig cells can be scaled up to meet demands in a highly standardised manner, and with respect to animal welfare regulations; iv) ‘designated-pathogen-free’ (DPF) pig lines can be produced, which could result in a higher safety profile than allotransplantation itself; v) the risk of zoonosis, which was raised years ago as the major hurdle, has been recently circumvented and is actually viewed as a controlled risk; and vi) immune risks are being circumvented via the use of genetically modified donor animals and encapsulation of porcine cells, particularly for the treatment of diabetes. Overall, the benefit appears to outweigh potential risks with respect to cellular xenotransplantation and this is discussed further in this review

A survey on mineral and heavy metal composition of Brazilian grape juice, wine, and sparkling wine produced from grapes cultivated under sustainable viticulture.

Grape growing and winemaking are important social and economical activines in Serra Gaúcha, which is located in Rio Grande do Sul, the Brazilian southernmost state.Resumo S14.295

Italicum e riforma costituzionale, verso un cambiamento della forma di governo?

Obiettivo di questa tesi è sostenere come la forma di governo parlamentare italiana stia progressivamente virando verso un presidenzialismo di fatto demarcato da un progressivo rafforzarsi di istituti che rafforzano l'esecutivo ed il Governo ponendolo al centro del sistema

Il rinvio pregiudiziale e l’indipendenza dei giudici: alcune riflessioni a margine di due recenti vicende

This essay focuses on the use of preliminary reference to the European Court of Justice pursuant to art. 267 TFEU in order to protect the independence and autonomy of National judiciaries, in particular within Member States such as Hungary and Poland. In the first part, the author looks at two specific cases in which the possibility for national judges to refer to the Court of Justice was limited by the action of the two illiberal governments. Secondly, also in relation to the decisions of the Brussels Court on the subject, the analysis focuses on the need to protect the preliminary reference in the context of judicial cooperation and on the role of European institutions, in particular the Commission, in doing so.This essay focuses on the use of preliminary reference to the European Court of Justice pursuant to art. 267 TFEU in order to protect the independence and autonomy of National judiciaries, in particular within Member States such as Hungary and Poland. In the first part, the author looks at two specific cases in which the possibility for national judges to refer to the Court of Justice was limited by the action of the two illiberal governments. Secondly, also in relation to the decisions of the Brussels Court on the subject, the analysis focuses on the need to protect the preliminary reference in the context of judicial cooperation and on the role of European institutions, in particular the Commission, in doing so

Frações do nitrogênio na microbiota de solos do sul do Brasil

The reaction of nitrogen compounds with ninhydrin can be used as an indicator of cytoplasmic materials released from microbial cells killed by fumigation. Total-N, ninhydrin-reactive-N (NR-N), ammonium-N (A-N), and a-amino-N in the microbial biomass of soils from the State of Rio Grande do Sul, Brazil, were determined, in 1996, in 0.5 mol L-1 K2SO4 extracts of fumigated and non-fumigated soils. Total-N varied from 20.3 to 104.4 mg kg-1 and the ninhydrinreactive- N corresponded, in average, to 27% of this. The ninhydrin-reactive-N was made up of 67% ammonium-N and 33% aminoacids with the amino group at the a-carbon position. It was concluded that colorimetric analysis of NR-N and A-N may be used as a direct measure of microbial N in soil. This simple and rapid procedure is adequate for routine analyses.A reação de compostos nitrogenados com a ninhidrina pode ser utilizada como parâmetro indicador dos produtos citoplásmicos oriundos da lise de células microbianas mortas pela fumigação. Com base nessa assertiva, realizou-se o presente trabalho, em 1996, com o objetivo de avaliar as concentrações de N-total, N-reativo com ninhidrina (NR-N), N-amônio (A-N) e N-a-amino presentes na biomassa microbiana de solos do Rio Grande do Sul. A partir da extração com K2SO4 0,5 mol L-1 de amostras provenientes de solos fumigados e não fumigados, determinaram-se as frações nitrogenadas presentes na biomassa. Constatouse que a quantidade do N-total presente na biomassa variou de 20,3 a 104,4 mg kg-1 de solo, sendo a fração microbiana reativa com a ninhidrina correspondente a 27% desses valores, em média. Dois terços dessa fração eram constituídos de nitrogênio na forma de amôniomicrobiano e um terço na forma de aminoácidos contendo o radical amino no carbono a. É possível utilizar a análise colorimétrica do NR-N e do A-N para a quantificação do N-microbiano do solo. Tais procedimentos são rápidos, simples e aplicáveis em análises de rotina

Comparative study of the interaction of CHAPS and Triton X-100 with the erythrocyte membrane

AbstractThe zwitterionic detergent CHAPS, a derivative of the bile salts, is widely used in membrane protein solubilization. It is a “facial” detergent, having a hydrophilic side and a hydrophobic back. The objective of this work is to characterize the interaction of CHAPS with a cell membrane. To this aim, erythrocytes were incubated with a wide range of detergent concentrations in order to determine CHAPS partition behavior, and its effects on membrane lipid order, hemolytic effects, and the solubilization of membrane phospholipids and cholesterol. The results were compared with those obtained with the nonionic detergent Triton X-100. It was found that CHAPS has a low affinity for the erythrocyte membrane (partition coefficient K=0.06mM−1), and at sub-hemolytic concentrations it causes little effect on membrane lipid order. CHAPS hemolysis and phospholipid solubilization are closely correlated. On the other side, binding of Triton X-100 disorders the membrane at all levels, and has independent mechanisms for hemolysis and solubilization. Differential behavior was observed in the solubilization of phospholipids and cholesterol. Thus, the detergent resistant membranes (DRM) obtained with the two detergents will have different composition. The behaviors of the two detergents are related to the differences in their molecular structures, suggesting that CHAPS does not penetrate the lipid bilayer but binds in a flat position on the erythrocyte surface, both in intact and cholesterol depleted erythrocytes. A relevant result for Triton X-100 is that hemolysis is not directly correlated with the solubilization of membrane lipids, as it is usually assumed

Fósforo remanescente para determinar a disponibilidade de fósforo em solos do Rio Grande do Sul

The objective of this work was to evaluate remaining P compared with soil clay content as a P buffer index to classify P extracted by the Mehlich-1 (M1) and Mehlich-3 (M3) methods in soils from the state of Rio Grande do Sul, Brazil. The experiment was carried out in a completely randomized design with five P2O5 rates (0, 50, 100, 200, and 400 mg kg-1) and two sucessive corn crops, and three replicates, in 20 representative soils of the state. P extracted by M1 and M3 before crop planting was adjusted to P contents in biomass, considering soil buffer capacity. The division of soils into different buffering classes, based on soil clay or remaining P, improved the capacity of estimating soil available P of both methods. However, there was no difference among the correlation coefficients obtained by classifying soils according to the evaluated indexes (remaining P or soil clay) for both M1 and M3 methods. Remaining P is a viable alternative to replace soil clay content to classify soil P extracted with the M1 and M3 methods.O objetivo deste trabalho foi avaliar o P remanescente comparado ao teor de argila como índice tampão para classificar o P extraído pelos métodos de Mehlich-1 (M1) e Mehlich-3 (M3), em solos do Rio Grande do Sul. O experimento foi realizado em delineamento inteiramente casualisado, com cinco doses de P2O5 (0, 50, 100, 200 e 400 mg kg-1), dois cultivos sucessivos de milho, e três repetições, em 20 solos representativos do Estado. O P extraído pelos métodos M1 e M3, antes dos cultivos, foi ajustado aos conteúdos de P na biomassa, tendo-se considerado a capacidade tampão do solo. A divisão dos solos em classes de tamponamento, de acordo com o teor de argila ou com o P remanescente, melhorou a capacidade preditiva do P disponível para ambos os métodos. Todavia, não houve diferença entre os coeficientes de correlação obtidos pela classificação dos solos de acordo com os índices avaliados (P remanescente ou teor de argila), tanto para o método M1 como para o M3. A análise do P-rem é uma alternativa viável para substituir o teor de argila na classificação do P extraído pelos métodos M1 e M3