The use of plasma cell-free DNA (cfDNA) as a useful biomarker in obstetric clinical practice has been delayed due to the lack of reliable quantification protocols. We developed a protocol to quantify ...plasma cfDNA using an internal standard strategy to overcome difficulties posed by low levels and high fragmentation of cfDNA. cfDNA was isolated from plasma samples of non-pregnant (NP, n = 26) and pregnant (P, n = 26) women using a commercial kit and several elution volumes were evaluated. qPCR parameters were optimized for cfDNA quantification, and several quantities of a recombinant standard were evaluated as internal standard. Absolute quantification was performed using a standard curve and the quality of the complete method was evaluated. cfDNA was eluted in a 50-μl volume, actin-β (ACTB) was selected as the target gene, and qPCR parameters were optimized. The ACTB standard was constructed and 1000 copies were selected as internal standard. The standard curve showed R
= 0.993 and E = 109.7%, and the linear dynamic range was defined between 10
and 10
ACTB copies/tube. Repeatability and reproducibility in terms of CV were 19% and up to 49.5% for ACTB copies per milliliter of plasma, respectively. The range of cfDNA levels was 428-18,851 copies/mL in NP women and 4031-2,019,363 copies/mL in P women, showing significant differences between the groups. We recommend the application of internal standard strategy for a reliable plasma cfDNA quantification. This methodology holds great potential for a future application in the obstetric field.
The influence of the cheese-making process, ripening conditions and primary starter on the viability and proteolytic activity of an adjunct culture of Lactobacillus plantarum I91 was assessed in two ...miniature cheese models, representative of Cremoso Argentino and Cheddar cheeses. Cheeses with and without adjunct culture were made under controlled microbiological conditions and sampled during ripening for physicochemical and microbiological analyses. The addition of lactobacilli neither contributed to acid production nor caused changes to the composition of the cheeses. The strain studied exhibited good development and survival and showed a similar growth pattern in both cheese matrices. The adjunct culture caused changes to secondary proteolysis of both cheese types, which were evidenced by modification of peptide profiles and the increase in the levels of some individual amino acids as well as the total content of free amino acids. The changes observed were consistent with the acceleration of proteolysis in the two cheese models assayed. Lactobacillus plantarum I91 has desirable and robust technological properties, which makes it a suitable adjunct culture for cheese-making. Other cultures and environmental conditions prevailing in the food may affect the viability of adjunct cultures and its biochemical activities; this is the first report describing the successful performance of an adjunct culture of Lact. plantarum I91 in two different model cheese systems.
Punzi-loss Abudinén, F.; Bertemes, M.; Bilokin, S. ...
The European physical journal. C, Particles and fields,
2022/2, Letnik:
82, Številka:
2
Journal Article
Recenzirano
Odprti dostop
We present the novel implementation of a non-differentiable metric approximation and a corresponding loss-scheduling aimed at the search for new particles of unknown mass in high energy physics ...experiments. We call the loss-scheduling, based on the minimisation of a figure-of-merit related function typical of particle physics, a Punzi-loss function, and the neural network that utilises this loss function a Punzi-net. We show that the Punzi-net outperforms standard multivariate analysis techniques and generalises well to mass hypotheses for which it was not trained. This is achieved by training a single classifier that provides a coherent and optimal classification of all signal hypotheses over the whole search space. Our result constitutes a complementary approach to fully differentiable analyses in particle physics. We implemented this work using PyTorch and provide users full access to a public repository containing all the codes and a training example.
This paper focuses on the importance of taking into account scattering of material parameters (zinc in this example) to perform sheet metal forming simulations. The approach described in this paper ...is efficiently pragmatic to consider as inputs the material parameters obtained during the industrial quality process. Large amount of collected data enables to obtain statistical evolution of the material law parameters and to determine afterwards the variability domain which is the final goal of this work. In order to get these parameters, an automatic calculation of material parameters (consistency, hardening coefficient and stress criterion for the formability as described by Jansen et al. (2013)) is developed in this work. Tensile tests have been performed in three directions (0°, 45° and 90°) and a Lagrangian interpolation was used for other orientations. In addition, scattering of the entire forming limit diagram is obtained from bulge tests performed for a sufficient statistical number of sheets. Based on this data, a numerical model integrating the scattering parameters was developed to build a statistical prediction of the stress failure criterion. All parameters of the material behavior law can be associated to the local calculations of the stress, strain, strain rate and temperature provided by a standard deterministic FEM computation and therefore giving a view of the failure rate of the process. This approach has been confronted to an industrial case describing the first forging pass of an elbow.
In this work, we studied the growth, survival, and peptidolytic activity of Lactobacillus plantarum I91 in a hard-cheese model consisting of a sterile extract of Reggianito cheese. To assess the ...influence of the primary starter and initial proteolysis level on these parameters, we prepared the extracts with cheeses that were produced using 2 different starter strains of Lactobacillus helveticus 138 or 209 (Lh138 or Lh209) at 3 ripening times: 3, 90, and 180d. The experimental extracts were inoculated with Lb. plantarum I91; the control extracts were not inoculated and the blank extracts were heat-treated to inactivate enzymes and were not inoculated. All extracts were incubated at 34°C for 21d, and then the pH, microbiological counts, and proteolysis profiles were determined. The basal proteolysis profiles in the extracts of young cheeses made with either strain tested were similar, but many differences between the proteolysis profiles of the extracts of the Lh138 and Lh209 cheeses were found when riper cheeses were used. The pH values in the blank and control extracts did not change, and no microbial growth was detected. In contrast, the pH value in experimental extracts decreased, and this decrease was more pronounced in extracts obtained from either of the young cheeses and from the Lh209 cheese at any stage of ripening. Lactobacillus plantarum I91 grew up to 8 log during the first days of incubation in all of the extracts, but then the number of viable cells decreased, the extent of which depended on the starter strain and the age of the cheese used for the extract. The decrease in the counts of Lb. plantarum I91 was observed mainly in the extracts in which the pH had diminished the most. In addition, the extracts that best supported the viability of Lb. plantarum I91 during incubation had the highest free amino acids content. The effect of Lb. plantarum I91 on the proteolysis profile of the extracts was marginal. Significant changes in the content of free amino acids suggested that the catabolism of free amino acids by Lb. plantarum I91 prevailed in a weakly proteolyzed medium, whereas the release of amino acids due to peptidolysis overcame their catabolism in a medium with high levels of free amino acids. Lactobacillus plantarum I91 was able to use energy sources other than lactose to support its growth because equivalent numbers of cells were observed in extracts containing residual amounts of lactose and in lactose-depleted extracts. The contribution of Lb. plantarum I91 to hard-cooked cheese peptidolysis was negligible compared with that of the starter strain; however, its ability to transform amino acids is a promising feature of this strain.
Punzi-loss Abudinén, F; Bertemes, M; Bilokin, S ...
European physical journal. C, Particles and fields,
02/2022, Letnik:
82, Številka:
2
Journal Article
Recenzirano
Odprti dostop
We present the novel implementation of a non-differentiable metric approximation and a corresponding loss-scheduling aimed at the search for new particles of unknown mass in high energy physics ...experiments. We call the loss-scheduling, based on the minimisation of a figure-of-merit related function typical of particle physics, a Punzi-loss function, and the neural network that utilises this loss function a Punzi-net. We show that the Punzi-net outperforms standard multivariate analysis techniques and generalises well to mass hypotheses for which it was not trained. This is achieved by training a single classifier that provides a coherent and optimal classification of all signal hypotheses over the whole search space. Our result constitutes a complementary approach to fully differentiable analyses in particle physics. We implemented this work using PyTorch and provide users full access to a public repository containing all the codes and a training example.
Hard cooked cheeses are mostly manufactured with lactic starters of
Lactobacillus helveticus, which constitute a major proteolytic agent in the food. In this work, we assessed the proteolysis ...produced by enzymes of two strains of
L. helveticus in a new cheese model, which consisted of a sterile substrate prepared with hard-cooked cheeses, and identified the time of ripening when main changes in proteolysis are produced. The extract, a representative model of the aqueous phase of the cheeses, was obtained from Reggianito cheeses of different ripening times (3, 90, and 180
days) made with starters composed of the strains tested, either SF138 or SF209. To obtain the substrate, the cheese was extracted with water, then centrifuged and the aqueous phase was sterilized by filtration through membrane (0.45
μm). The substrates were incubated at 34
°C during 21
days; samples were taken at 0, 3, 7, 14, and 21
days. Sterility was verified by plating samples on skim milk agar and incubating at 37
°C for 48
h. Proteolysis was determined by liquid chromatography of soluble peptides and free amino acids. Great variation in peptide profiles was found as incubation progressed in cheese extracts, which evidenced that proteases and peptidases from the starter were active and able to degrade the proteinaceous material available in the extracts. The extracts derived from cheeses with
L. helveticus SF138 showed low production of peptides and a notable increase in free amino acids content during incubation.
L. helveticus SF209, on the contrary, caused an increase on soluble peptides, but the free amino acids accumulation was lower than in the first case, which suggested that
L. helveticus SF209 had either a low peptydolitic activity or produced an intense amino acids breakdown. This trend was more evident for extracts prepared with 90-day-old cheeses. It was concluded that the strains of
L. helveticus assayed showed potentially complementary proteolytic abilities, as SF209 was able to provide a continuous replenishment of peptides during incubation, while SF138 increased their hydrolysis to free amino acids. The extract was an appropriate medium to model hard cooked cheese ripening in short periods of time.
Self-heating of rolled ZnCuTi sheets MILESI, M; MUÑOZ, D PINO; LAGROUM, A ...
Sadhana (Bangalore),
12/2020, Letnik:
45, Številka:
1
Journal Article
Recenzirano
Self-heating effect for thermomechanical forming processes turns out to be dramatically significant for material with a very low melting point, such as zinc alloys. Zinc low melting point temperature ...(419.54°C) accentuates metallurgical effects when it is formed in cold or warm manufacturing processes. During deformation, self-heating can therefore induce a relative softening in the behavior law (that could improve formability) which competes with the conventional plastic hardening of the material. Hence, the correct identification of zinc material behavior has to account for both softening and hardening phenomena. This paper studies the effect of plastic work energy in the material identified by means of tensile tests by combining digital image correlation and infrared thermography technique.
► Neonatal exposure to endosulfan altered uterine morphoregulatory proteins. ► Endosulfan at doses similar to AID and NOEL might exhibit estrogenic activity. ► We are investigating if neonatal ...exposure to endosulfan affects uterine functions.
This study investigates the effects of neonatal exposure to low doses of endosulfan on the expression of proteins regulating uterine development and differentiation. Female pups received vehicle, endosulfan (Endo6: 6μg/kg, Endo600: 600μg/kg) or diethylstilbestrol (DES: 0.2μg/kg) from postnatal day 1 (PND1) to PND7. The uterine expression of estrogen receptor alpha (ERα), progesterone receptor (PR), Hoxa10 and alpha smooth muscle actin (α-SMA) was detected by immunohistochemistry on PND8 (neonatal period) and PND21 (prepubertal period), to evaluate acute and short-term responses. ERα, Hoxa10 and α-SMA were induced in the Endo600 group in both ages, while a striking decrease in PR expression was detected in the prepubertal rats following each dose of endosulfan. DES treatment deregulated ERα and Hoxa10 uterine expression at each age. Studies are currently underway to investigate whether the dysregulation of steroid receptors, Hoxa10 and α-SMA observed following neonatal exposure to endosulfan affect uterine functions in adulthood.
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