Chronic hepatitis B virus infection (CHB) caused by the hepatitis B virus (HBV) is one of the most common viral infections in the world. Reactivation of HBV infection is a life-threatening condition observed in patients with CHB receiving chemotherapy or other medications. Although HBV reactivation is commonly attributed to immune suppression, other factors have long been suspected to play a role, including intracellular signaling activated in response to DNA damage. We investigated the effects of DNA-damaging factors (doxorubicin and hydrogen peroxide) on HBV reactivation/replication and the consequent DNA-damage response. Dose-dependent activation of HBV replication was observed in response to doxorubicin and hydrogen peroxide which was associated with a marked elevation in the mRNA levels of ataxia-telangiectasia mutated (ATM) and ATM- and RAD3-related (ATR) kinases. Downregulation of ATM or ATR expression by shRNAs substantially reduced the levels of HBV RNAs and DNA. In contrast, transcriptional activation of ATM or ATR using CRISPRa significantly increased HBV replication. We conclude that ATM and ATR are essential for HBV replication. Furthermore, DNA damage leading to the activation of ATM and ATR transcription, results in the reactivation of HBV replication.

Maize tortilla is a basic food in Mexico, and, lately, the food industry has tried to make the manufacturing process easier by using instant flours and specialized machines. The purpose of this study was to investigate consumers' behaviors related to tortillas and to evaluate the sensory, textural, and physico-chemical parameters of tortillas from the Tlazala region, Mexico. The sensory profile revealed that the artisanal ones had better parameters in terms of smell, taste, and appearance compared to the others. These results are consistent with consumers' preferences for tortillas made of maize grain instead of industrial corn flour. The sensory parameters and the physico-chemical and texture profile parameters varied with the maize type and manufacturing process. Our findings showed that the artisanal hand-made ones were more nutritious, followed by those mechanically made using maize grain, and finally by those mechanically made from industrialized corn flour. The results of this study may help processors to better understand the parameters of their products and people's preferences.

The bioactivity of ten traditional Korean Angelica species were screened by angiotensin-converting enzyme (ACE) assay in vitro. Among the crude extracts, the methanol extract of Angelica decursiva whole plants exhibited potent inhibitory effects against ACE. In addition, the ACE inhibitory activity of coumarins 1-5, 8-18 was evaluated, along with two phenolic acids (6, 7) obtained from A. decursiva. Among profound coumarins, 11-18 were determined to manifest marked inhibitory activity against ACE with IC50 values of 4.68-20.04 µM. Compounds 12, 13, and 15 displayed competitive inhibition against ACE. Molecular docking studies confirmed that coumarins inhibited ACE via many hydrogen bond and hydrophobic interactions with catalytic residues and zinc ion of C- and N-domain ACE that blocked the catalytic activity of ACE. The results derived from these computational and in vitro experiments give additional scientific support to the anecdotal use of A. decursiva in traditional medicine to treat cardiovascular diseases such as hypertension.

Studies on the relationship between reactive oxygen species (ROS)/manganese superoxide dismutase (MnSOD) and sphingomyelinase (SMase) are controversial. It has been demonstrated that SMase increases the intracellular ROS level and induces gene expression for MnSOD protein. On the other hand, some authors showed that ROS modulate the activation of SMase. The human recombinant manganese superoxide dismutase (rMnSOD) exerting a radioprotective effect on normal cells, qualifies as a possible pharmaceutical tool to prevent and/or cure damages derived from accidental exposure to ionizing radiation. This study aimed to identify neutral SMase (nSMase) as novel molecule connecting rMnSOD to its radiation protective effects. We used a new, and to this date, unique, experimental model to assess the effect of both radiation and rMnSOD in the brain of mice, within a collaborative project among Italian research groups and the Joint Institute for Nuclear Research, Dubna (Russia). Mice were exposed to a set of minor γ radiation and neutrons and a spectrum of neutrons, simulating the radiation levels to which cosmonauts will be exposed during deep-space, long-term missions. Groups of mice were treated or not-treated (controls) with daily subcutaneous injections of rMnSOD during a period of 10 days. An additional group of mice was also pretreated with rMnSOD for three days before irradiation, as a model for preventive measures. We demonstrate that rMnSOD significantly protects the midbrain cells from radiation-induced damage, inducing a strong upregulation of nSMase gene and protein expression. Pretreatment with rMnSOD before irradiation protects the brain with a value of very high nSMase activity, indicating that high levels of activity might be sufficient to exert the rMnSOD preventive role. In conclusion, the protective effect of rMnSOD from radiation-induced brain damage may require nSMase enzyme.

This paper proposes a convenient route to oxidize the -CH2-OH groups in the water-soluble pullulan, using a new catalytic polymer-supported N-hydroxyphthalimide (NHPI) immobilized on polystyrene. The protocol involves the presence of sodium hypochlorite and sodium bromide. The conversion is possible at room temperature, atmospheric pressure, and pH = 10. The characterization of both the catalysts and oxidized pullulan was done using NMR and FTIR methods. Using polyelectrolyte titration with end-point indication by means of a particle-charge detector (PCD), we were able to assess the degree of electrokinetic charge in all oxidized samples as a consequence of the conversion of the -CH2-OH group into -COOH moieties. The possibility of recovery and recycling of the polymer-supported NHPI catalyst was tested for up to four cycles, since the morphological analyses performed on the catalysts using SEM revealed no significant changes.

Tyrosine kinase inhibitors (TKI) such as the BCR-ABL inhibitor dasatinib and nilotinib are highly effective therapies for chronic myeloid leukemia (CML). However, several lines of evidence suggest that dasatinib can induce bleeding which may be due to impaired collagen-induced platelet adhesion, aggregation, and secretion. Sarcoma family kinases (SFK) play central role in the GPVI-induced signaling pathway. We aimed to investigate whether and how dasatinib can modulate SFK-mediated platelet procoagulant activity in a purified system and in dasatinib/nilotinib treated CML patients. In platelet rich plasmas of healthy volunteers, dasatinib dose-dependently reduced convulxin-induced phosphatidylserine exposure and attenuated thrombin formation. Similarly to these changes, integrin activation and clot retraction were also significantly inhibited by 100 nM dasatinib. Platelets isolated from dasatinib treated patients showed a significantly lower phosphatidylserine expression upon convulxin activation compared to premedication levels. In these samples, thrombin generation was significantly slower, and the quantity of formed thrombin was less compared to the trough sample. Western blot analyses showed decreased phosphorylation levels of the C-terminal tail and the activation loop of SFKs upon dasatinib administration. Taken together, these results suggest that dasatinib inhibits the formation of procoagulant platelets via the GPVI receptor by inhibiting phosphorylation of SFKs.

Approximately 20% of children with hepatoblastoma (HB) have metastatic disease at diagnosis, most frequently in the lungs. In children with HB, lung metastatic disease is associated with poorer prognosis. Its treatment has been approached with a variety of methods that integrate chemotherapy and surgical resection. The timing and feasibility of complete extirpation of lung metastases, by chemotherapy and/or metastasectomy, is crucial for the surgical treatment of the primary liver tumor, which can vary from major hepatic resections to liver transplantation (LT). In children with unresectable HB, which can be surgically treated only by LT, the persistence of unresectable metastases after neoadjuvant chemotherapy excludes the possibility of recurring to LT with consequent negative impact on patients' outcomes. Due to limited evidence and experience, there is no consensus amongst oncologists and surgeons across institutions regarding the surgical treatment for HB with synchronous metastatic lung disease. This narrative review aimed to update the current management of pulmonary metastasis in children with HB and to define its role in the decision-making strategy for the surgical approach to primary liver tumours.

As a novel natural compound delivery system, liposomes are capable of incorporating lipophilic bioactive compounds with enhanced compound solubility, stability and bioavailability, and have been successfully translated into real-time clinical applications. To construct the soy phosphatidylcholine (SPC)-cholesterol (Chol) liposome system, the optimal formulation was investigated as 3:1 of SPC to Chol, 10% mannosylerythritol lipid-A (MEL-A) and 1% betulinic acid. Results show that liposomes with or without betulinic acid or MEL-A are able to inhibit the proliferation of HepG2 cells with a dose-effect relation remarkably. In addition, the modification of MEL-A in liposomes can significantly promote cell apoptosis and strengthen the destruction of mitochondrial membrane potential in HepG2 cells. Liposomes containing MEL-A and betulinic acid have exhibited excellent anticancer activity, which provide factual basis for the development of MEL-A in the anti-cancer applications. These results provide a design thought to develop delivery liposome systems carrying betulinic acid with enhanced functional and pharmaceutical attributes.

Dp71 is the most abundant dystrophin (DMD) gene product in the nervous system. Mutation in the Dp71 coding region is associated with cognitive disturbances in Duchenne muscular dystrophy (DMD) patients, but the function of dystrophin Dp71 in tumor progression remains to be established. This study investigated Dp71 expression in glioblastoma, the most common and aggressive primary tumor of the central nervous system (CNS).

The feeding regime of early, supplementary solid diet improved rumen development and production in goat kids. However, the signature microbiota responsible for linking dietary regimes to rumen function shifts are still unclear. This work analyzed the rumen microbiome and functions affected by an early solid diet regime using a combination of machine learning algorithms. Volatile fatty acids (i.e., acetate, propionate and butyrate) fermented by microbes were found to increase significantly in the supplementary solid diet groups. Predominant genera were found to alter significantly from unclassified Sphingobacteriaceae (non-supplementary group) to Prevotella (supplementary solid diet groups). Random Forest classification model revealed signature microbiota for solid diet that positively correlated with macronutrient intake, and linearly increased with volatile fatty acid production. Bacteria associated with carbohydrate and protein metabolism were also identified. Utilization of a Fish Taco analysis portrayed a set of intersecting core species contributed to rumen function shifts by the solid diet regime. The core community structures consisted of the specific, signature microbiota and the manipulation of their symbiotic partners are manipulated by extra nutrients from concentrate and/or forage, and then produce more volatile fatty acids to promote rumen development and functions eventually host development. Our study provides mechanisms of the microbiome governed by a solid diet regime early in life, and highlights the signature microbiota involved in animal health and production.

The membrane phosphoproteome in plant seed changes dynamically during embryo development. We examined the patterns of Phaseolus vulgaris (common bean) seed membrane protein phosphorylation from the mid-maturation stage until two days after germination. Serine and threonine phosphorylation declined during seed maturation while tyrosine phosphorylation remained relatively constant. We discovered that the aquaporin PvTIP3;1 is the primary seed membrane phosphoprotein, and PvTIP3;2 shows a very low level of expression. The level of phosphorylated Ser7 in PvTIP3;1 increased four-fold after seed maturation. Since phosphorylation increases water channel activity, we infer that water transport by PvTIP3;1 is highest in dry and germinating seeds, which would be optimal for seed imbibition. By the use of isoform-specific, polyclonal peptide antibodies, we found that PvTIP3;2 is expressed in a developmental pattern similar to PvTIP3;1. Unexpectedly, PvTIP3;2 is tyrosine phosphorylated following seed maturation, which may suggest a mechanism for the regulation of PvTIP3;2 following seed germination. Analysis of protein secondary structure by circular dichroism spectroscopy indicated that the amino-terminal domain of PvTIP3;1 is generally unstructured, and phosphorylation increases polyproline II (PPII) helical structure. The carboxy-terminal domain also gains PPII character, but in a pH-dependent manner. These structural changes are a first step to understand TIP3 aquaporin regulation.

Phenotypic plasticity and local adaptation via genetic change are two major mechanisms of response to dynamic environmental conditions. These mechanisms are not mutually exclusive, since genetic change can establish similar phenotypes to plasticity. This connection between both mechanisms raises the question of how much of the variation observed between species or populations is plastic and how much of it is genetic. In this study, we used a structured population of fire salamanders (Salamandra salamandra), in which two subpopulations differ in terms of physiology, genetics, mate-, and habitat preferences. Our goal was to identify candidate genes for differential habitat adaptation in this system, and to explore the degree of plasticity compared to local adaptation. We therefore performed a reciprocal transfer experiment of stream- and pond-originated salamander larvae and analyzed changes in morphology and transcriptomic profile (using species-specific microarrays). We observed that stream- and pond-originated individuals diverge in morphology and gene expression. For instance, pond-originated larvae have larger gills, likely to cope with oxygen-poor ponds. When transferred to streams, pond-originated larvae showed a high degree of plasticity, resembling the morphology and gene expression of stream-originated larvae (reversion); however the same was not found for stream-originated larvae when transferred to ponds, where the expression of genes related to reduction-oxidation processes was increased, possibly to cope with environmental stress. The lack of symmetrical responses between transplanted animals highlights the fact that the adaptations are not fully plastic and that some level of local adaptation has already occurred in this population. This study illuminates the process by which phenotypic plasticity allows local adaptation to new environments and its potential role in the pathway of incipient speciation.

Ferrocenyl-functionalized thioketones have recently been recognized as useful building blocks for sulfur-containing compounds with potential applications in materials chemistry. This work is devoted to a single representative of such thioketones, namely diferrocenyl thioketone (Fc2CS), whose structure has been determined here for the first time. Both X-ray crystallography and a wide variety of quantum-chemical methods were used to explore the structure of Fc2CS. In addition to the X-ray structure determination, intermolecular interactions occurring in the crystal structure of Fc2CS were examined in detail by quantum-chemical methods. These methods were also an invaluable tool in studying the molecular structure of Fc2CS, from the gas phase to solutions and to its crystal. Intramolecular interactions governing the conformational behavior of an isolated Fc2CS molecule were deduced from quantum-chemical analyses carried out in orbital space and real space. Our experimental and theoretical results indicate that the main structural features of an isolated Fc2CS molecule in its lowest-energy geometry are retained both upon solvation and in the crystal. The tilt of ferrocenyl groups is only slightly affected by crystal packing forces that are dominated by dispersion. Nonetheless, a network of intermolecular interactions, such as H···H, C···H and S···H, was detected in the Fc2CS crystal but each of them is fairly weak.

Considering the implications of adverse effects of chemotherapy (CT) and the potential impact of diet on patients' recovery, this study aimed to prospectively evaluate the association between the consumption of food groups, patients' Dietary Inflammatory Index (DII®) scores, and their nutritional status. Anthropometric and dietary assessments of 55 women with breast cancer (BC) were performed at three time points. T0 is the time point after the first CT cycle, T1 is the time point after the intermediate CT cycle, and T2 is the time point after the last CT cycle. We identified a significant increase in weight, body mass index, and waist circumference during CT. Consumption of poultry and eggs was higher in T1 when compared to T2, while consumption of total fruit and total vegetables was higher at T0 compared to T1 and T2. The diet became more pro-inflammatory over the course of treatment (X2(2) = 61.127), and was related to higher abdominal adiposity. Total fruit (T0: R2 = 0.208, T1: R2 = 0.095, T2: R2 = 0.120) and total vegetable consumption (T0: R2 = 0.284, T1: R2 = 0.365, T2: R2 = 0.580) predicted DII® change at the three-time points. Meanwhile, consumption of total grains was significantly associated only with T1 (R2 = 0.084) and T2 (R2 = 0.118), and consumption of simple sugars was significantly associated only with T0 (R2 = 0.137) and T1 (R2 = 0.126). Changes in food consumption led to an increase in the inflammatory profile of the diet, suggesting the necessity to improve the guidelines during and after CT. These results reinforce the need to promote healthier eating practices in concert with maintaining a healthy nutritional status in women with BC treated with CT.

Ectropis grisescens and Ectropis obliqua are sibling species of tea-chewing pests. An investigation of the distribution of tea geometrids was implemented for enhancing controlling efficiency. E. grisescens is distributed across a wider range of tea-producing areas than Ectropis obliqua in China with sympatric distribution found in some areas. In order to explore reproductive isolation mechanisms in co-occurrence areas, hybridization experiments were carried out. Results showed they can mate but produce infertile hybrids. During experiments, the desynchronized development phenomenon was found in the hybridized generation of sibling tea geometrids. Furthermore, transcriptome analysis of those individuals of fast-growing and slow-growing morphs revealed that the insect hormone biosynthesis pathway was enriched in two unsynchronized development groups of hybrid offspring. More importantly, some genes regulating the synthesis of moulting hormone showed significantly up-regulated expression in fast-growing groups. Above all, metabolism of the juvenile hormone and synthesis of the ecdysone pathway were found to be crucially involved in the desynchronized development phenomenon. This research finding contributes to a better understanding of the mechanisms of insect development and reproductive isolation of two sibling species.

Methyl benzoate (MB) is a component of bee semiochemicals. Recent discovery of insecticidal activity of MB against insect pests provides a potential alternative to chemical insecticides. The aim of this study was to examine any potential adverse impact of MB on honey bees. By using two different methods, a spray for contact and feeding for oral toxicity, LC50s were 236.61 and 824.99 g a.i./L, respectively. The spray toxicity was 2002-fold and 173,163-fold lower than that of imidacloprid and abamectin. Piperonyl butoxide (PBO, inhibiting P450 oxidases [P450]) significantly synergized MB toxicity in honey bees, indicating P450s are the major MB-detoxification enzymes for bees. Assessing additive/synergistic interactions indicated that MB synergistically or additively aggravated the toxicity of all four insecticides (representing four different classes) in honey bees. Another adverse effect of MB in honey bees was the significant decrease of orientation and flight ability by approximately 53%. Other influences of MB included minor decrease of sucrose consumption, minor increase of P450 enzymatic activity, and little to no effect on esterase and glutathione S-transferase (GST) activities. By providing data from multiple experiments, we have substantially better understanding how important the P450s are in detoxifying MB in honey bees. MB could adversely affect feeding and flight in honey bees, and may interact with many conventional insecticides to aggravate toxicity to bees. However, MB is a relatively safe chemical to bees. Proper formulation and optimizing proportion of MB in mixtures may be achievable to enhance efficacy against pests and minimize adverse impact of MB on honey bees.

Mesenchymal stem cells have been widely implicated in tumour development and metastases. Moving from the use of two-dimensional (2D) models to three-dimensional (3D) to investigate this relationship is critical to facilitate more applicable and relevant research on the tumour microenvironment. We investigated the effects of altering glucose concentration and the source of foetal bovine serum (FBS) on the growth of two breast cancer cell lines (T47D and MDA-MB-231) and human bone marrow-derived mesenchymal stem cells (hBM-MSCs) to determine successful conditions to enable their co-culture in 3D tumour spheroid models. Subsequently, these 3D multi-cellular tumour spheroids were used to investigate the effect of hBM-MSCs on breast cancer cell invasiveness. Findings presented herein show that serum source had a statistically significant effect on two thirds of the growth parameters measured across all three cell lines, whereas glucose only had a statistically significant effect on 6%. It was determined that the optimum growth media composition for the co-culture of 3D hBM-MSCs and breast cancer cell line spheroids was 1 g/L glucose DMEM supplemented with 10% FBS from source A. Subsequent results demonstrated that co-culture of hBM-MSCs and MDA-MB-231 cells dramatically reduced invasiveness of both cell lines (F(1,4) = 71.465, p = 0.001) when embedded into a matrix comprising of growth-factor reduced base membrane extract (BME) and collagen.

The antimicrobial activity of natural antimicrobials (fruit extracts, essential oils and derivates), was assessed against six bacteria species (E. coli O157:H7, L. monocytogenes, S. Typhimurium, B. subtilis, E. faecium and S. aureus), two molds (A. flavus and P. chrysogenum) and a yeast (C. albicans) using disk diffusion method. Then, the antimicrobial compounds having high inhibitory capacity were evaluated for the determination of their minimum inhibitory, bactericidal and fungicidal concentration (MIC, MBC and MFC respectively). Total phenols and flavonoids content, radical scavenging activity and ferric reducing antioxidant power of selected compounds were also evaluated. Based on in vitro assays, five antimicrobial compounds were selected for their lowest effective concentration. Results showed that, most of these antimicrobial compounds had a high concentration of total phenols and flavonoids and a good anti-oxidant and anti-radical activity. In situ study showed that natural antimicrobials mix, applied on the carrot surface, reduced significantly the count of the initial mesophilic total flora (TMF), molds and yeasts and allowed an extension of the shelf-life of carrots by two days as compared to the control. However, the chemical treatment (mix of peroxyacetic acid and hydrogen peroxide) showed antifungal activity and a slight reduction of TMF.

This study examined the association between peritoneal cytology and survival in early-stage cervical cancer. This is a nationwide multicenter retrospective study, examining consecutive women with clinical stage IB1-IIB cervical cancer who underwent radical hysterectomy with available peritoneal cytology results from 2004-2008. Propensity score inverse probability of treatment weighting was used to assess the impact of malignant peritoneal cytology on survival. Among 1409 analyzed cases, 88 (6.2%) had malignant peritoneal cytology. On weighted models, malignant peritoneal cytology was associated with decreased disease-free survival (hazard ratio (HR) 1.78, 95% confidence interval (CI) 1.36-2.32) and overall survival (OS, HR 1.93, 95% CI 1.44-2.59). On sensitivity analyses, malignant peritoneal cytology was associated with decreased OS in adenocarcinoma/adenosquamous carcinoma, high-risk early-stage disease and those who received concurrent chemo-radiotherapy. However, among women who received postoperative systemic chemotherapy, malignant peritoneal cytology was not associated with OS (HR 1.21, 95% CI 0.72-2.04). A systematic review, including our results, showed that malignant peritoneal cytology was associated with decreased OS (HR 4.03, 95% CI 1.81-8.99) and increased recurrence in squamous carcinoma (odds ratio 1.89, 95% CI 1.05-3.39) and adenocarcinoma (odds ratio 4.30, 95% CI 2.30-8.02). In conclusion, the presence of malignant cells in peritoneal cytology is associated with decreased survival in early-stage cervical cancer. The possible benefit of systemic chemotherapy in this subgroup merits further investigation.

Microglia respond to adverse stimuli in order to restore brain homeostasis and, upon activation, they release a number of inflammatory mediators. Chronic microglial overactivation is related to neuroinflammation in Alzheimer's disease. In this work, we show that oleanolic acid (OA), a natural triterpene present in food and medicinal plants, attenuates the activation of BV2 microglial cells induced by lipopolysaccharide (LPS). Cell pretreatment with OA inhibited the release of IL-1β, IL-6, TNF-α, and NO, which was associated with the downregulation of the expression of genes encoding for these cytokines and inducible nitric oxide synthase (iNOS), and the reinforcement of the endogenous antioxidant cell defense. These findings advocate considering OA as a novel neuroprotective agent to inhibit oxidative stress and inflammatory response in activated microglia associated with Alzheimer's disease.