Omacetaxine may have a role in chronic myeloid leukaemia eradication through downregulation of Mcl-1 and induction of apoptosis in stem/progenitor cells

Chronic myeloid leukaemia (CML) is maintained by a rare population of tyrosine kinase inhibitor (TKI)-insensitive malignant stem cells. Our long-term aim is to find a BcrAbl-independent drug that can be combined with a TKI to improve overall disease response in chronic-phase CML. Omacetaxine mepesuccinate, a first in class cetaxine, has been evaluated by clinical trials in TKI-insensitive/resistant CML. Omacetaxine inhibits synthesis of anti-apoptotic proteins of the Bcl-2 family, including (myeloid cell leukaemia) Mcl-1, leading to cell death. Omacetaxine effectively induced apoptosis in primary CML stem cells (CD34<sup>+</sup>38<sup>lo</sup>) by downregulation of Mcl-1 protein. In contrast to our previous findings with TKIs, omacetaxine did not accumulate undivided cells <i>in vitro</i>. Furthermore, the functionality of surviving stem cells following omacetaxine exposure was significantly reduced in a dose-dependant manner, as determined by colony forming cell and the more stringent long-term culture initiating cell colony assays. This stem cell-directed activity was not limited to CML stem cells as both normal and non-CML CD34<sup>+</sup> cells were sensitive to inhibition. Thus, although omacetaxine is not leukaemia stem cell specific, its ability to induce apoptosis of leukaemic stem cells distinguishes it from TKIs and creates the potential for a curative strategy for persistent disease

Modelling autocatalytic behaviour of a food model system- sucrose thermal degradation at high concentrations

Sucrose thermal degradation is an important reaction in the food industry. When in concentrated and neutral solutions, pure sucrose presents a lag phase in the reaction. This work aimed at: (i) modelling sucrose thermal degradation autocatalytic behaviour and (ii) studying the concentration and temperature effects on kinetic parameters. Isothermal experiments were conducted at temperatures ranging from 100 to 180 C, using solutions with varying water content (3.58–30.03 (% w/w)). The logistic and Gompertz sigmoidal equations were modified and reparameterised, in order to describe degradation behaviour with kinetic parameters with physical meaning (maximum reaction rate, kmax, and lag time, k). In both models these parameters presented an Arrhenius type dependence on temperature. Following a mixed model effect methodology, the concentration dependence was observed on the Arrhenius parameters. This concentration effect was included in the proposed kinetic models, which were able to successfully describe experimental data

International survey of Cronobacter sakazakii and other Cronobacter spp. in follow up formulas and infant food

A coordinated survey for Cronobacter and related organisms in powdered infant formula, follow up formula and infant foods was undertaken by 8 laboratories in 7 countries in recognition of and in response to the data needs identified in an FAO/WHO call for data in order to develop global risk management guidance for these products. The products (domestic and imported) were purchased from the local market and were categorised according to their principle ingredients. A total of 290 products were analysed using a standardised procedure of pre-enrichment in 225 ml Buffered Peptone Water (BPW), followed by enrichment in Enterobacteriaceae Enrichment (EE) broth, plating on the chromogenic Cronobacter Druggan–Forsythe–Iversen (DFI) agar and presumptive identification with ID 32 E. Presumptive Cronobacter isolates were identified using 16S rRNA gene sequence analysis. Aerobic plate counts (APC) of the products were also determined on nutrient agar. Fourteen samples had APC > 105 cfu/g, 3 of which contained probiotic cultures. C. sakazakii was isolated from 27 products; 3/91 (3%) follow up formulas (as defined by Codex Alimentarius Commission), and 24/199 (12%) infant foods and drinks. Hence C. sakazakii was less prevalent in follow up formula than other foods given to infants over the same age range. A range of other bacteria were also isolated from follow up formulas, including Acinetobacter baumannii, Enterobacter cloacae, Klebsiella pneumoniae, Citrobacter freundii, and Serratia ficaria. There was significant variation in the reconstitution instructions for follow up formulas. These included using water at temperatures which would enable bacterial growth. Additionally, the definition of follow up formula varied between countries

Modelling colour changes during the caramelisation reaction

Sucrose solutions, with concentrations near or superior to saturation, present high potentialities for the candy and pastry industries. The development of colour in a neutral and highly concentrated sucrose solution (16.32%(w/w) water content) subjected to isothermal heat treatment (in the 100–160 C range) was investigated. Under such conditions, sucrose degrades through caramelisation and 5- hydroxymethylfurfural (HMF) is formed. Colour development was monitored through lightness/darkening (L/L0) and total colour difference (TCDH) changes during reaction course. Kinetic behaviour was mathematically described using modified Gompertz equations. The effect of temperature on the reaction was described by an Arrhenius type dependency. Colour development and sucrose degradation kinetic parameters were compared and similar lag phases were found. However, the same was not observed for reactions rate, indicating that not only sucrose degradation contributes to colour development. To investigate the colour development/HMF content relationship, a fractional conversion and a power law models where successfully proposed to express, respectively, L/L0 and TCDH dependence on HMF content

Mathematical modelling of viscosity near the glass transition: the random-walk approach

The present work deals with the application of the random-walk model, proposed by Arkhipov and Bässler, to describe the viscosity dependence on temperature. Data obtained for sucrose solutions were used to assess the validity of the random-walk approach and the normality of the Density of Possible Metastable States (DPMS). Good results for the fitting of the proposed models were found for fragile liquids. Strong liquids showed poor results and require more investigation

Performance evaluation of deleteriousness prediction methods for intronic SNVs in next generation sequences

Introduction: Alterations in splicing sites (ss) are estimated to explain approximately 10% of human disease causal variants. Mutations outside the ss but affecting ?regulatory elements? can be up to 25%. Accurate deleteriousness prediction for intronic variants is crucial for diagnostic purposes. Many deleteriousness prediction methods have been developed, but their relative values are still unclear in practical applications. We comprehensively evaluated the predictive performance of two complementary deleteriousness-scoring methods using information from real patients. Material and Methods: We selected the dbscSNV (both ADA and RF scores) and SPIDEX algorithms, that study variants in splicing consensus regions or in regulatory regions respectively. The tools, either alone or in combination, were tested on 29294 gene intronic SNVs that have previously been characterised by ClinVar as either ?pathogenic? (430) or ?benign? (28864). The sensitivity, specificity and positive and negative predictive values were calculated. Moreover, we applied the algorithms to WES data from undiagnosed patients, and we analysed the mRNA sequence from genes that fitted the patient?s phenotype. Results: The highest sensitivity corresponds to dbscSNV with 96.55% while the best specificity is for SPIDEX with 95.78%. When considering the 3 scores (SPIDEX, dbscSNV ADA and RF Score), the sensitivity and specificity values were 60.7% and 94.6%. The Positive and Negative Predictive Value were 14.45% and 99.39%. The results for 20 undiagnosed cases are presented. Conclusions: Besides the low positive predictive value, the combination of both algorithms leads less than 1% of false negatives, so their routine use can be recommended for diagnostic purposes

Innovating portuguese traditional pastry – on the use of pasteurised egg yolk in “ovos moles”

“Ovos moles” is a renowned traditional Portuguese sweet that is confectioned with egg yolk, sugar and water. In this work a safer and easier to manipulate raw material - pasteurised liquid egg yolk - is proposed to substitute the intact shell eggs used in traditional “ovos moles” production. Due to previous heat treatment, pasteurised liquid egg yolk presents different sensorial, nutritional and physical properties. These changes may alter the final product’s characteristics. In order to develop a safer formulation, using pasteurised raw products, with minimised differences from the traditional product, a study on adding pasteurised egg white (as bulking agent) to the pasteurised liquid yolk was carried out. Samples made with normal shell eggs and with pasteurised liquid eggs were compared in terms of rheological parameters. Results allow characterising the physical properties of traditional ovos moles and of alternative formulations, using pasteurised egg yolk and white. A new formula, with rheological properties identical to the traditional one, was developed