Heterologous production of 3-hydroxy-lanosta-8, 24-dien-26 oic acid (HLDOA) was recently achieved by expressing CYP5150L8 from Ganoderma lucidum in Saccharomyces cerevisiae, but post-modification of HLDOA remains unclear. In this study, another P450 from G. lucidum, CYP5139G1, was identified to be responsible for C-28 oxidation of HLDOA, resulting in the formation of a new ganoderic acid (GA) 3,28-dihydroxy-lanosta-8, 24-dien-26 oic acid (DHLDOA) by the engineered yeast, whose chemical structure was confirmed by LC-MS and NMR. In vitro enzymatic experiments confirmed the oxidation of HLDOA to DHLDOA by CYP5139G1. As the DHLDOA production was low (0.27 mg/L), to improve it, the strategy of adjusting the dosage of hygromycin and geneticin G418 to respectively manipulate the copy number of plasmid pRS425-Hyg-CYP5150L8-iGLCPR (harboring CYP5150L8, iGLCPR and hygromycin resistant gene hygR) and pRS426-KanMx-CYP5139G1 (harboring CYP5139G1 and G418 resistant gene KanMx) was adopted. Finally, 2.2 mg/L of DHLDOA was obtained, which was 8.2 fold of the control (without antibiotics addition). The work not only enriches the library of GAs and GA biosynthetic enzymes, but also helps to construct heterologous cell factories for other GA production as well as to elucidate the authentic GA biosynthetic pathway in G. lucidum.

Many salmonid species exist in highly structured and isolated populations, and are susceptible to habitat fragmentation and disturbances. Gila Trout (Oncorhynchus gilae) is a threatened species found in the Southwestern United States. Gila trout is managed to preserve remnant populations (i.e. lineages) distributed across a fragmented landscape. We evaluated genomic variation within and among remaining lineages of Gila Trout using RADseq to assess how drift and selection have structured populations using neutral and outlier loci. We also examined whether a signature of hybridization was evident in relict populations. Despite Gila Trout lineages being significantly differentiated and highly structured with low effective population sizes, we found that most lineages maintained genomic diversity and were potentially locally adapted. Hybridization with non-native Rainbow Trout (O. mykiss) was not detected in any lineage. Some lineages may have experienced recent population bottlenecks perhaps associated with mortality from drought and severe wildfires. Current management strategies should be reevaluated and adapted to better account for long-term effects of climate change. Specifically, we suggest reconnecting some populations via dendritic stream networks to facilitate natural dispersal in a metapopulation context. This would allow natural genetic mixing on the landscape and potentially increase adaptive potential. Furthermore, genetic rescue should be implemented to preserve integrity of the unique Spruce Creek lineage that is currently compromised by extremely low diversity.

Introduction: muscle pain/fibromyalgia (FM) is common among individuals with myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). We recently demonstrated that during orthostatic stress testing, adults with ME/CFS reported increased pain. In the current study, we hypothesized that pain pressure thresholds (PPT) would decrease and temporal summation (windup) would increase after head-up tilt testing (HUT), and that the presence of co-morbid FM would be associated with greater change in both measures. Methods and Results: 248 ME/CFS patients (164 with FM and84 without FM), and 22 healthy controls (HC) were analyzed. In HC there were no significant differences in PPT between pre- and post-HUT (finger: from 4.7(1.6) to 4.4(1.5); shoulder: from 2.8(1.0) to 2.9(1.0)). In ME/CFS patients with and without FM, a significant decrease in PPT post-HUT was found compared to HC (both p<0.0001). Patients with FM had a lower PPT pre- and post-HUT (finger: from 2.0(0.9) to 1.5(0.8); shoulder: from 1.2(0.5) to 1.0(0.5) compared to patients without FM (finger: from 5.0(1.6) to 3.3(1.5); shoulder: from 2.2(0.9) to1.9(1.0) (p ranging from 0.001 to <0.0001). In contrast to HC in ME/CFS patients windup was increased compared to HC pre-HUT (both p<0.0001), but did not significantly change post-HUT. Conclusions: Pressure pain threshold decreased in ME/CFS patients with or without fibromyalgia after head-up tilt test (HUT), compared to healthy controls. Windup pre-and post-HUT was significantly higher compared to healthy controls, but did not change from pre- to post-HUT. These results demonstrate that, like exercise, orthostatic stress can negatively influence the physiology of pain perception in ME/CFS.

This work aimed to test innovations for the diagnosis of agricultural properties at Rio do Peixe Watershed, to locate erosions and to correct them by changing soil management, aiming at recovering degraded areas, rehabilitating them and promoting conservation agriculture, evaluating the results by remote sensing and water quality indicators. In 2019, it turned twelve years of inspection/monitoring at Rio do Peixe Watershed. From 2007 to 2017, using the Conventional CDA methodology, 14,076 ha were inspected at Vera Cruz sector, 94 properties were notified and in Ocauçu, 82 properties, located in 9,027 ha. As a work strategy, in Marília, the Innovative CDA Methodology was used, which allowed the inspection and rehabilitation of 52 properties in 27,775 ha, from 2017 to 2018. After the notifications, the owners presented the conservationist technical projects for each property, which were implemented, using conservation practices such as improving vegetation cover and crop rotation to control laminar erosion and agricultural terracing, divergent channels and containment basin to control gullies erosions. This work promoted a transformation from degrading agriculture to conservation agriculture, having degraded pastures transformed into an agricultural area, implementing the No-tillage system. Pastures were recovered by implementing the Integrated Crop-Livestock System; it was possible to increase the occupancy rate by 31% comparing to the original situation. This is a great work that benefits Watershed farmers, increasing productivity and consequently the profit, as well as for the local people improving the quality of water that supplies the region of Marília.

Background: Early extubation after cardiac surgery results in better postoperative outcomes but the optimal time for extubation remains unclear. Premature extubation may lead to unplanned reintubation that may result in adverse outcomes. The present study was undertaken to compare preoperative and operative risks and outcomes of patients with prolonged intubation >48 hours and reintubation after cardiac surgery. Aims: This is a retrospective chart review of 1259 patients who had cardiac surgery at a community hospital. After excluding patients with either operating room death, extubation <48 hours without reintubation or extubation >48 hours with reintubation, the final sample included 83 patients (6.6%) requiring reintubation after extubation in <48 hours and 100 (8.0%) with prolonged intubation without requiring reintubation. Results: Bivariate analyses revealed few statistically different preoperative and operative risks between patients with reintubation and prolonged intubation. Reintubation patients were older (p = .033) and had lower body mass index (p = .000), higher preoperative hematocrit (p = .021), and more chronic kidney disease stages >2 (p = .046) but lower odds for intra-aortic balloon pump (p = .006) and emergency surgery (p = .005). Reintubation led to worse postoperative outcomes than prolonged intubation: more acute kidney injury stage 1-3 (p = .014), coma/encephalopathy (p = .004), postoperative transfusion (p = .003), increased intensive care unit length of stay (p = .001) and hospital mortality (p = .007). Based on binary and ordinal logistic regression analyses, the differences in preoperative and operative risks were either inconsistent or trivial contributors and reintubation appears to make the largest independent contribution to poor postoperative outcomes. Conclusion: While early extubation remains the goal, patients with marginal weaning readiness may benefit from more recovery time before extubation. This study suggests that premature extubation may increase unplanned reintubation that could adversely affect postoperative outcomes after cardiac surgery.

Vitamin D insufficiency is closely related to various kinds of metabolic diseases. Acted as a marker of vitamin D status, 25-HydroxyvitaminD3 detection possesses important practical significance. In this study, highly sensitive fluorescent detection of 25-HydroxyvitaminD3 using truncated affinity-improved aptamers were developed, based on fluorescence intensity changes of PicoGreen (PG) generated upon binding to double-stranded DNA (dsDNA) formed through hybridization of aptamer and corresponding complementary strand. Four truncated aptamers were obtained by intercepting the small hairpin loop as the functional domain and retaining double helix structural domains of different lengths that exist in the selected original 25-HydroxyvitaminD3 aptamer. Under the optimized PG concentration, we conducted comparison experiments for affinity and specificity of these four truncated aptamers. Among them, the shortest aptamer with only 21 bp, D3-4, was found to show the highest affinity and specificity to 25-HydroxyvitaminD3, with the limit of detection of 0.04μg/mL, which solved the problem that original long aptamer could not applied for this fluorescent detection of 25-HydroxyvitaminD3. The truncated 25-HydroxyvitaminD3-specific aptamer with highly enhanced affinity performs promising application in sensitive detection of 25-HydroxyvitaminD3.

Methods This prospective observational study included 44 patients who underwent isolated ASD closures in the department of cardiac surgery at the National Heart Foundation Hospital and Research Institute (NHFH & RI) from July 2014 to June 2016. After fulfilling the enrolment criteria, participants were divided into two groups.

Tissue engineering (TE) constitutes a multidisciplinary scientific discipline focused on the construction of artificial tissues to regenerate end-stage organs. The impact of TE has led to a clinical revolution since novel therapies are available to attend several conditions. In this sense, its onset has supposed the communication of innovative discoveries in the age of social and mass media. This study aims to evaluate the global online dimension of TE from 2012 to 2018 by using data from the Web of Science (WoS) and Altmetrics. We have analysed 23,719 documents through descriptive and statistical methodologies. First, the descriptive analysis showed the evolution of TE original articles in five online platforms (Twitter, Patents, Facebook, Mendeley readers and News) and compared the most relevant TE documents ranked by their traditional and alternative metrics of impact. Secondly, we carried out a correlation and factorial analysis and then constructed a linear regression model to define a mathematical equation for the prediction of future TE citations counts from Altmetric scores. The obtained results suggest a growing presence of TE in the online social web and the feasibility in the context of global science to anticipate TE traditional academic impact by using social media

We report the case of a man affected by atopic dermatitis on dupilumab. During the treatment, the patient showed a worsening of head and neck dermatitis. Blue light irradiation is a safe and efficacy tool for head and neck dermatitis persisting or developed during therapy with dupilumab.

After surgical removal of bone tumors, elimination of the remains of cancer cells along with tissue healing and functionality is a therapeutic goal. Regarding the destructive effect of chemo-/radiotherapy on healthy cells, the development of multimodal scaffolds with simultaneous anticancer and osteo-regenerative potency is of particular importance as regenerative medicine for bone tissue engineering. Our previous study demonstrated that hollow pollen grain (HPG) of Pistacia vera L. offers a unique bone-forming activity and encapsulating capacity that it can be considered as an excellent scaffolding building block in bottom-up bone engineering. In the present study, for the first time, the anticancer potential of Pistacia vera L. HPG was investigated in-vitro using human osteosarcoma cell line MG63. Optical imaging of the HPG-cell interface indicated large focal adhesion due to the HPG unique surface features such as functionality and ornamentation. MTT assay results showed an anti-proliferative effect of HPG on the MG63 cells. Moreover, from the gene expression and DAPI staining analyses, HPG of Pistacia vera L. was found to be an apoptosis-inducing building block against MG63 bone cancer cells. So therefore, such a multifunctional building block with the ability of discriminatory killing human osteosarcoma cell line is proposed to be used after surgery to not only hinder cancer recurrence after surgery but also to stimulate bone healing.

Antimicrobial photodynamic therapy (APDT) is a promising alternative to traditional antibiotics for bacterial infections, which inactivates a broad spectrum of bacteria. However, it has some disadvantages including poor water solubility and easy aggregation of hydrophobic photosensitizers (PS), and poor tissue penetration and cytotoxicity when using UV as light source, leading to photodynamic therapy efficacy. Herein, we develop a novel water-soluble natural PS (sorbicillinoids) obtained by microbial fermentation using filamentous fungus Trichoderma reesei (T. reesei). Sorbicillinoids could effectively generate singlet oxygen (1O2) under UV light irradiation, and ultimately display photoinactivation activity on Gram-positive bacteria, but not Gram-negative ones. Staphylococcus aureus (S. aureus) treated with sorbicillinoids and UV light displayed high levels of intracellular reactive oxygen species (ROS), notable DNA photocleavage, and compromised membrane permeability without overt cell membrane disruption. Moreover, the dark toxicity, phototoxicity or hemolysis activity of sorbicillinoids is negligible, showing its excellent biocompatibility.

Pressure ulcers are commonly associated with microbial infections on the wounds which need an effective wound dressing. However, the silver dressings have shown promising result but they have toxicity and argyria. Hence, this study aimed to develop and characterize chitosan-polyethylene glycol (PEG) nanocomposite hydrogel loaded with phomopsidione as an antimicrobial dressing. The hydrogel being synthesized was analyzed with transmission and scanning electron microscopes. Drug release and mechanical properties were studied having confirmed the functional groups with Fourier transform infrared (FTIR) spectroscopy. Finally, antimicrobial activities were evaluated against the clinical wound pathogens. The developed hydrogel was soft, flexible and elastic, having nanospheres of chitosan-PEG but no sign of aggregation under the electron microscopes. Releasing of phomopsidione from the nanocomposite hydrogel was slow and gradual following the first order of kinetic. On average, 34 μg/mL phomopsidione released per hour and 67.9% active ingredients delivered into the surrounding medium over the study period. Although, the bioactivity activity of the hydrogel was narrow-spectrum, it showed significant results against all Gram-negative bacteria and Candida utilis with 99.99% reduction of microbial growth. The findings reveal that the phomopsidione loaded hydrogel has a great promise to act as an antimicrobial dressing for chronic wounds.

A document by Laipubam Sharma . Click on the document to view its contents.

Early detection of Hepatitis B virus (HBV) in pregnant females starts by a request of the gynecologists, which is based on their knowledge and awareness of the guidelines on conducting these tests. This is an observational cross-sectional study that investigated the gynecologists’ knowledge, attitude and practice regarding HBV during pregnancy across Jordan. A random sample of 150 gynecologists were approached, from 3 major cities in Jordan, and asked to fill a questionnaire that assessed their knowledge, attitude and practice. Statistical analysis was conducted using SPSS. One hundred and seven gynecologists have participated in the study. Most of the respondents were females, residents, and less than thirty years old. Although 97.2% of the practitioners agreed on the importance of the pregnant females on HBV, only 43% were aware of the obligatory protocol in Jordan regarding HBV screening, and only 55% would screen the pregnant females to HBV in-practice. Significant association was found between screening rates to HBV and both, level of specialty and experience of practitioners. Approximately 60% of practitioners were aware of HBV perinatal transmission risk. Only 19.6% always referred the infected pregnant females to other specialists regardless of the viral load. While 47.7% of practitioners agreed on recommending antiviral therapy for third trimester pregnant females, only 12.1% would always/often prescribe them. A noteworthy lack of Hepatitis B knowledge and screening practice among gynecologists in Jordan have been observed. A national program designed to increase the awareness of HBV testing in Jordan for both patients and gynecologists is called for.

Oxygen availability and overflow metabolism are often limiting factors in high cell-density cultures. In the present study, expression of Vitreoscilla hemoglobin in the chromosome of Escherichia coli was used as a strategy to improve plasmid DNA (pDNA) production in biphasic fed-batch cultures. During the fully aerobic batch phase, the strain expressing VHb accumulated 28 % less acetate and 19 % more pDNA than the non-expressing strain. The fed-batch phase was carried out with a change of regime from aerobic to microaerobic conditions. The pDNA yields from biomass increased consistently in the VHb-expressing strain during the whole culture, while decreased progressively for the non-expressing strain during microaerobic conditions. The ratio of positive/negative plasmid replication control molecules (RNAII/RNAI) were lower for the VHb-expressing than for the non-expressing strain. However, the final pDNA titer was ca. 74 % higher for the former. Flux balance analysis suggests that VHb presence increases the flux in anaplerotic pathways. The higher lactate production observed in VHb-expressing cells may be triggered by an increased demand of NAD+ in glycolysis under microaerobic conditions. These results are valuable for faster development of robust pDNA production processes.

In this theoretical study, we investigate the electronic potential energy curves, spectroscopic parameters, vibrational energy levels and transition dipole moments for the diatomic dications BeRb2+, BeCs2+ and SrRb2+. We consider an ab initio approach based on the use of non-empirical pseudopotentials and parameterized l dependent polarization potentials. Results show that 1-22Σ+ for BeRb2+, 1-52Σ+ for BeCs2+ and 1-32Σ+ for SrRb2+ are repulsive. While the 32Σ+ for BeRb2+, 42Σ+ for BeCs2+ and 42Σ+ for SrRb2+ are metastable states. These states can accommodate some vibrational energy levels. Interesting avoided crossings between some 2+ states are localized and examined. Until now no experimental and theoretical studies have been made for each system. Consequently, we discuss our results by comparing with some data of similar systems. Besides, the transition dipole moments of the ground state to a few excited states are computed and presented. The information associated with the electronic structures, spectroscopic parameters as well as the transition properties that provide in this paper is anticipated to serve as guidelines for further experimental and theoretical researches for each diatomic dication considered in this work.

Urea hydrolysis is widely used in agriculture, environment and other engineering fields, among which urease contained in beans can catalyze urea hydrolysis. The urea hydrolysis activity of legume plant leaching solution(LPLS) was investigated, including soybeans, black beans, mung beans, red beans as well as soybean hulls, soybean leaves, soybean stems and soybean pods. For the high urea hydrolysis activity and economic efficiency, soybean is most suitable for agricultural engineering and other fields than other beans and soybean-related parts extract. The urea hydrolysis activity increases with the concentration of LPLS, while decreases gradually with reaction time. When the heating temperature reaches 25, 35, 45, 55 and 65 ℃, the urea hydrolysis activity is steady and the enzyme activity is high. Enzyme activity decreases after 65 ° C (i.e.75, 90 ° C). Meanwhile, the soaking time of LPLS has a little effect on the urea hydrolysis activity compared with other factors. These results make a positive contribution to domestic production urease experimental basis.

Background Application of genetically modified non-pathogenic bacteria expressing specific anti-tumor proteins under certain conditions specific to tumors is an effective approach for selective targeting of tumors. We developed here, for the first time, a novel spatiotemporal cancer targeted therapy applying engineered E. coli bacteria with capability of expressing cardiac peptides under hypoxic conditions of tumor. Method E. coli BW25133 was transformed with construction of co-expressing cardiac hormones and GFP. Bacteria bearing constructs were then IV administered in mice bearing tumors and then tumor localization, as well as tumor proliferation, invasion and angiogenesis biomarkers (Ki-67, VEGFR, CD31and MMP9), changes in cytokine profile, suppression of tumor growth and survival were analyzed. Results IV Administered bacteria bearing constructs could specifically localize at tumor site and express cardiac peptides under hypoxic conditions. Administration of bacteria significantly enhanced survival rate, suppressed tumor progression and lowered expression levels of MMP-9, VEGFR2, CD31and Ki67 as potent markers for angiogenesis, tumor proliferation and metastasis. Furthermore, applied bacteria resulted in significant reduction in the expression of IL-1β, IL-6, GC-SF, IL-12 and TNF-α proinflammatory cytokines, whereas increasing IL-10, IL-17A and INF-γ cytokines. Conclusion Overall, administration of E. coli bearing cardiac hormone expression construct could effectively suppress tumor growth, angiogenesis, invasion and metastasis while enhancing survival rate in mice model of breast cancer.