BlockProof: A Framework for Verifying Authenticity and Integrity of Web Content.

In the Literature, we can find several research works to help in the digital crime fight in order to prove integrity and authenticity of a published document, image or video. Among all the crimes, fake news certainly is among the most recurrent ones and needs to be mitigated. There are several Blockchain-based applications in order to make use of the benefits derived from technology, but little is found to verify the authenticity of Web content records as well as the history of all updates that have taken place in each Web content. Such kind of solution has become important nowadays as a way to cover the gap in the combat against fake news, for example. The purpose of this paper is to present BlockProof, a framework for verifying web content authenticity and integrity that offers a solution for content providers to register Web content, regardless of whether the page has dynamic or static content, in addition to enabling the consultation of the history of all records made for a given URL. We understand that such kind of solution may be useful to data producers/providers to provide evidence that they are in compliance with the fight against fake news, for instance.

Predicting Popularity of Video Streaming Services with Representation Learning: A Survey and a Real-World Case Study.

The Internet's popularization has increased the amount of content produced and consumed on the web. To take advantage of this new market, major content producers such as Netflix and Amazon Prime have emerged, focusing on video streaming services. However, despite the large number and diversity of videos made available by these content providers, few of them attract the attention of most users. For example, in the data explored in this article, only 6% of the most popular videos account for 85% of total views. Finding out in advance which videos will be popular is not trivial, especially given many influencing variables. Nevertheless, a tool with this ability would be of great value to help dimension network infrastructure and properly recommend new content to users. In this way, this manuscript examines the machine learning-based approaches that have been proposed to solve the prediction of web content popularity. To this end, we first survey the literature and elaborate a taxonomy that classifies models according to predictive features and describes state-of-the-art features and techniques used to solve this task. While analyzing previous works, we saw an opportunity to use textual features for video prediction. Thus, additionally, we propose a case study that combines features acquired through attribute engineering and word embedding to predict the popularity of a video. The first approach is based on predictive attributes defined by resource engineering. The second takes advantage of word embeddings from video descriptions and titles. We experimented with the proposed techniques in a set of videos from GloboPlay, the largest provider of video streaming services in Latin America. A combination of engineering features and embeddings using the Random Forest algorithm achieved the best result, with an accuracy of 87%.

Renal sympathetic activation triggered by the rostral ventrolateral medulla is dependent of spinal cord AT1 receptors in Goldblatt hypertensive rats.

Spinal cord neurons contribute to elevated sympathetic vasomotor activity in renovascular hypertension (2K1C), particularly, increased actions of angiotensin II. However, the origin of these spinal angiotensinergic inputs remains unclear. The present study aimed to investigate the role of spinal angiotensin II type 1 receptor (AT1) receptors in the sympathoexcitatory responses evoked by the activation of the rostral ventrolateral medulla (RVLM) in control and 2K1C Goldblatt rats. Hypertension was induced by clipping of the left renal artery. After 6 weeks, a catheter (PE-10) filled with losartan was inserted into the subarachnoid space and advanced to the T10-11 vertebral level in urethane-anesthetized rats. The effects of glutamate microinjection into the RVLM on blood pressure (BP), heart rate (HR), and renal and splanchnic sympathetic nerve activity (rSNA and sSNA, respectively) were evaluated in the presence or absence of spinal AT1 blockade. Tachycardic, pressor, and renal sympathoexcitatory effects caused by RVLM activation were significantly blunted by losartan in 2K1C rats, but not in control rats. However, no differences were found in the gene expression of angiotensin-converting enzyme, angiotensinogen, and renin in the spinal cord segments between the groups. In conclusion, acute sympathoexcitation induced by RVLM activation is dependent on the spinal AT1 receptor in Goldblatt, but not in control, rats. The involvement of other central cardiovascular nuclei in spinal angiotensinergic actions, as well as the source of angiotensin II, remains to be determined in the Goldblatt model.

Mobile vs. Non-Mobile Live-Streaming: A Comparative Analysis of Users Engagement and Interruption Using Big Data from a Large CDN Perspective.

Video streaming on the Internet is constantly changing and growing. New devices and new video delivery mechanisms generate huge gaps in the understanding of how video application works. From exploratory research of one among the largest streaming services in Brazil, this work presents a comparison between mobile and non-mobile users, in large-scale lives. This work focuses on metrics such as engagement, interruption, churn, and payload. This work also presents a report-overview of mobile-users, considering the operating system, geolocation, network access, interruption, and engagement. These results might offer potential information for streaming improvement, in addition to serving as a historical mark.

Increased sympathetic responses induced by chronic obstructive sleep apnea are caused by sleep fragmentation.

Obstructive sleep apnea (OSA) is often associated with sympathetic overactivity and hypertension. These associations are mainly attributed to hypoxia acting on arterial chemoreceptors. However, the contribution of arousal from sleep is unclear. We measured the effect of OSA and sleep fragmentation on cardiovascular and sympathetic function and gene expression in the brain in rats. Male Wistar rats were fitted with a tracheal balloon and EEG and electromyogram electrodes and assigned to control (n = 6), OSA (n = 9), or arousal (n = 8) treatments. The OSA group was subjected to obstructive apnea, each time the rat entered sleep, for 8 h/day for 15 days. The arousal group was similarly exposed to vibration, which was produced with a miniature vibration motor mounted on the rat's head. Vibration intensity slowly increased until the rat awoke. One day after the last apnea or arousal, rats were anesthetized and arterial blood pressure and splanchnic sympathetic nerve activity (SSNA) were recorded. Baseline mean and diastolic pressure were increased after OSA. Resting SSNA was similar in the three groups, but both OSA and sleep fragmentation increased sympathetic activation in response to airway obstruction and chemoreflex activation by cyanide. OSA increased superoxide dismutases 1 and 2 in the brainstem, whereas sleep fragmentation did not. Our results suggest that sympathetic overactivity to chemoreceptor stimulation was a consequence of arousal from sleep. Our study suggests that sleep disruption may have an important role in the development of apnea-related sympathetic activation.NEW & NOTEWORTHY Obstructive sleep apnea causes a hyperactive chemoreflex, with increased sympathetic activation. However, it is not clear whether this pathophysiologic mechanism is due to repeated hypoxia or to sleep disruption. The present study suggests that sleep fragmentation contributes importantly to increased sympathetic activation after chemoreceptor stimulation. This suggests that sleep fragmentation has an important role in the sympathetic activation seen in sleep apnea patients.

Interaction between angiotensin II and GABA in the spinal cord regulates sympathetic vasomotor activity in Goldblatt hypertension.

Previous studies have been described changes in brain regions contributing to the sympathetic vasomotor overactivity in Goldblatt hypertension (2K1C). Furthermore, changes in the spinal cord are also involved in the cardiovascular and autonomic dysfunction in renovascular hypertension, as intrathecal (i.t.) administration of Losartan (Los) causes a robust hypotensive/sympathoinhibitory response in 2K1C but not in control rats. The present study evaluated the role of spinal γ-aminobutyric acid (GABA)-ergic inputs in the control of sympathetic vasomotor activity in the 2K1C rats. Hypertension was induced by clipping the renal artery. After six weeks, a catheter (PE-10) was inserted into the subarachnoid space and advanced to the T10-11 vertebral level in urethane-anaesthetized rats. The effects of i.t. injection of bicuculline (Bic) on blood pressure (BP), renal and splanchnic sympathetic nerve activity (rSNA and sSNA, respectively) were evaluated over 40 consecutive minutes in the presence or absence of spinal AT1 antagonism. I.t. Bic triggered a more intense pressor and sympathoexcitatory response in 2K1C rats, however, these responses were attenuated by previous i.t. Los. No differences in the gene expression of GAD 65 and GABA-A receptors subunits in the spinal cord segments were found. Thus, the sympathoexcitation induced by spinal GABA-A blockade is dependent of local AT1 receptor in 2K1C but not in control rats. Excitatory angiotensinergic inputs to sympathetic preganglionic neurons are tonic controlled by spinal GABAergic actions in Goldblatt hypertension.

Best practice for PTEN gene and protein assessment in anatomic pathology.

There is a lack of standardization of a best practice protocol for Phosphatase and Tensin Homolog (PTEN) assessment by immunohistochemistry in anatomic pathology routine practice. We performed immunohistochemistry for 19 antibodies against PTEN, eleven of which were excluded during the standardization step. Immunohistochemistry of the remaining eight antibodies was performed on a Tissue Microarray containing 55 prostate and 40 renal carcinoma samples. Fluorescent in situ hybridization (FISH) was used as reference standard for immunohistochemistry specificity evaluation. Concerning nuclear staining, polyclonal (Cat#22034-1-AP); 6H2.1 mMAb (Cat#ABM-2052), Y184 RabMAb (Cat#NB110-57441) and 217702 mMAb antibodies presented the highest agreement with fluorescent in situ hybridization (p<0.001 for all) and with regard to cytoplasmic staining, Y184 RabMAb (Cat#NB110-57441); polyclonal (Cat#22034-1-AP) and 217702 mMAb presented the highest agreement (p<0.001 for all). Our results indicate that several commercially available antibodies do not show reliability of sensitivity and specificity for PTEN evaluation and we propose 6H2.1 mMAb (Cat#ABM-2052) as the antibody of choice for laboratory standardization and best practice in clinical routine, which demonstrated excellent sensitivity for both nuclear and cytoplasmic staining, specificity for PTEN by Western blot and good correlation with PTEN status by FISH with regard to nuclear staining.

Eukaryotic initiation factor 5A dephosphorylation is required for translational arrest in stationary phase cells.

The protein known as eIF5A (eukaryotic initiation factor 5A) has an elusive role in translation. It has a unique and essential hypusine modification at a conserved lysine residue in most eukaryotes. In addition, this protein is modified by phosphorylation with unknown functions. In the present study we show that a phosphorylated state of eIF5A predominates in exponentially growing Trypanosoma cruzi cells, and extensive dephosphorylation occurs in cells in stationary phase. Phosphorylation occurs mainly at Ser(2), as shown in yeast eIF5A. In addition, a novel phosphorylation site was identified at Tyr(21). In exponential cells, T. cruzi eIF5A is partially associated with polysomes, compatible with a proposed function as an elongation factor, and becomes relatively enriched in polysomal fractions in stationary phase. Overexpression of the wild-type eIF5A, or eIF5A with Ser(2) replaced by an aspartate residue, but not by alanine, increases the rate of cell proliferation and protein synthesis. However, the presence of an aspartate residue instead of Ser(2) is toxic for cells reaching the stationary phase, which show a less-pronounced protein synthesis arrest and a decreased amount of eIF5A in dense fractions of sucrose gradients. We conclude that eIF5A phosphorylation and dephosphorylation cycles regulate translation according to the growth conditions.

Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion.

Protein tyrosine phosphatases (PTPs) form a large family of enzymes involved in the regulation of numerous cellular functions in eukaryotes. Several protein tyrosine phosphatases have been recently identified in trypanosomatides. Here we report the purification and biochemical characterization of TcPTP1, a protein tyrosine phosphatase from Trypanosoma cruzi, the causing agent of Chagas' disease. The enzyme was cloned and expressed recombinantly in Escherichia coli and purified by Ni-affinity chromatography. Biochemical characterization of recombinant TcPTP1 with the PTP pseudo-substrate pNPP allowed the estimation of a Michaelis-Menten constant K(m) of 4.5mM and a k(cat) of 2.8s(-1). We were able to demonstrate inhibition of the enzyme by the PTP1b inhibitor BZ3, which on its turn was able to accelerate the differentiation of epimastigotes into metacyclic forms of T. cruzi induced by nutritional stress. Additionally, this compound was able to inhibit by 50% the infectivity of T. cruzi trypomastigotes in a separate cellular assay. In conclusion our results indicate that TcPTP1 is of importance for cellular differentiation and invasivity of this parasite and thus is a valid target for the rational drug design of potential antibiotics directed against T. cruzi.

Polymorphism in the flanking regions of the PbGP43 gene from the human pathogen Paracoccidioides brasiliensis: search for protein binding sequences and poly(A) cleavage sites.

BACKGROUND: Paracoccidioides brasiliensis is a thermo-dimorphic fungus that causes paracoccidiodomycosis (PCM). Glycoprotein gp43 is the fungal main diagnostic antigen, which can also protect against murine PCM and interact with extracellular matrix proteins. It is structurally related to glucanases, however not active, and whose expression varies considerably. We have presently studied polymorphisms in the PbGP43 flanking regions to help understand such variations. RESULTS: we tested the protein-binding capacity of oligonucleotides covering the PbGP43 proximal 5' flanking region, including overlap and mutated probes. We used electrophoretic mobility shift assays and found DNA binding regions between positions -134 to -103 and -255 to -215. Only mutation at -230, characteristic of P. brasiliensis phylogenetic species PS2, altered binding affinity. Next, we cloned and sequenced the 5' intergenic region up to position -2,047 from P. brasiliensis Pb339 and observed that it is composed of three tandem repetitive regions of about 500 bp preceded upstream by 442 bp. Correspondent PCR fragments of about 2,000 bp were found in eight out of fourteen isolates; in PS2 samples they were 1,500-bp long due to the absence of one repetitive region, as detected in Pb3. We also compared fifty-six PbGP43 3' UTR sequences from ten isolates and have not observed polymorphisms; however we detected two main poly(A) clusters (1,420 to 1,441 and 1,451 to 1,457) of multiple cleavage sites. In a single isolate we found one to seven sites. CONCLUSIONS: We observed that the amount of PbGP43 transcripts accumulated in P. brasiliensis Pb339 grown in defined medium was about 1,000-fold higher than in Pb18 and 120-fold higher than in Pb3. We have described a series of features in the gene flanking regions and differences among isolates, including DNA-binding sequences, which might impact gene regulation. Little is known about regulatory sequences in thermo-dimorphic fungi. The peculiar structure of tandem repetitive fragments in the 5' intergenic region of PbGP43, their characteristic sequences, besides the presence of multiple poly(A) cleavage sites in the 3' UTR will certainly guide future studies.

Transcription regulation of the Pbgp43 gene by nitrogen in the human pathogen Paracoccidioides brasiliensis.

We show indirect evidences for the possible involvement of NIT2-like binding motifs in transcription modulation of the PbGP43 gene, which codes for an important antigen from the human fungal pathogen Paracoccidioides brasiliensis. This investigation was motivated by the finding of 23 NIT2-like sites within the proximal -2047 nucleotides of the PbGP43 5' intergenic region from the Pb339 isolate. They compose four clusters, two of them identical. We found four NIT2-containing probes that were positive in electrophoretic mobility shift assays and further analyzed them. PbGP43 could be modulated by nitrogen primary sources in Pb339, Pb3 and Pb18 isolates, as observed by reverse transcription (RT) real time-PCR. Gene reporter assays conducted in Aspergillus nidulans suggested that the minimal fragment responsible for nitrogen modulation lies within -480 bp of the PbGP43 gene. This is the first report on PbGP43 transcription modulation in response to nitrogen primary sources, which might help understand its regulation during infection.

Molecular karyotype analysis of Perkinsus atlanticus (Phylum Perkinsozoa) by pulsed field gel electrophoresis.

Perkinsus atlanticus is a pathogenic protist that infects the clam Ruditapes decussatus. Although it was recently proposed that the genus Perkinsus belongs to a new phylum, Perkinsozoa, in the infra-kingdom Alveolata, there remain different opinions about whether this genus should form a phylum on its own and consequently divergent views about its taxonomic characterization. In this work, we have identified nine chromosomes by pulsed field gel electrophoresis (PFGE) combined with densitometry analysis. The obtained karyotype of Perkinsus atlanticus, like that of other early branches of the dinoflagellate lineage, displays a more conventional chromosome organization, different from that of most dinoflagellates.