bluuebunny/crossref_metadata_2025 huggingface dataset:
1H NMR Based Metabolomics in Human Sepsis and Healthy Serum
Authors: Jaurila Henna , Koivukangas Vesa , Koskela Marjo , Gäddnäs Fiia , Myllymaa Sami , Kullaa Arja , Salo Tuula , Ala-Kokko Tero I.
Published: February 2020

Early diagnosis is essential but challenging in severe sepsis. Quantifying and comparing metabolite concentrations in serum has been suggested as a new diagnostic tool. Here we used proton nuclear magnetic resonance spectroscopy (1H NMR) based metabolomics to analyze the possible differences in metabolite concentrations between sera taken from septic patients and healthy controls, as well as between sera of surviving and non-surviving sepsis patients. We took serum samples from 44 sepsis patients when the first sepsis induced organ dysfunction was found. Serum samples were also collected from 14 age and gender matched healthy controls. The samples were analyzed by quantitative 1H NMR spectroscopy for non-lipid metabolites. We found that the serum levels of glucose, glycine, 3-hydroxybutyrate, creatinine and glycoprotein acetyls (mostly alpha-1-acid glycoprotein, AGP) were significantly (p < 0.05) higher in sepsis compared to healthy sera, whereas citrate and histidine were significantly (p < 0.05) lower in sepsis patients compared to healthy controls. We found statistically significantly higher serum lactate and citrate concentrations in non-survivors compared to 30-day survivors. According to our study, 3-hydroxybutyrate, citrate, glycine, histidine, and AGP are candidates for further studies to enable identification of phenotype association in the early stages of sepsis.
Pollution Gradients Altered the Bacterial Community Composition and Stochastic Process of Rural Polluted Ponds
Authors: Tai Xin , Li Rui , Zhang Bao , Yu Hao , Kong Xiao , Bai Zhihui , Deng Ye , Jia Lan , Jin Decai
Published: February 2020

Understanding the effects of pollution on ecological communities and the underlying mechanisms that drive them will helpful for selecting a method to mediate polluted ecosystems. Quantifying the relative importance of deterministic and stochastic processes is a very important issue in ecology. However, little is known about their effects on the succession of microbial communities in different pollution levels rural ponds. Also, the processes that govern bacterial communities in polluted ponds are poorly understood. In this study, the microbial communities in water and sediment from the ponds were investigated by using the 16S rRNA gene high-throughput sequencing technology. Meanwhile, we used null model analyses based on a taxonomic and phylogenetic metrics approach to test the microbial community assembly processes. Pollution levels were found to significantly alter the community composition and diversity of bacteria. In the sediment samples, the bacterial diversity indices decreased with increasing pollutant levels. Between-community analysis revealed that community assembly processes among water and sediment samples stochastic ratio both gradually decreased with the increased pollution levels, indicating a potential deterministic environmental filtering that is elicited by pollution. Our results identified assemblage drivers of bacterial community is important for improving the efficacies of ecological evaluation and remediation for contaminated freshwater systems.
Development of Crystalline Cu2S Nanowires via a Direct Synthesis Process and Its Potential Applications
Authors: Chen Chih-Yen , Jiang Jian-Ru , Chuang Wen-Shuo , Liu Ming-Song , Lee Sheng-Wei
Published: February 2020

Large-scale and uniform copper(I) sulfide (Cu2S) nanowires have been successfully synthesized via a cheap, fast, easily handled, and environmentally friendly approach. In addition to the reductive properties of the biomolecule-assisted method, they also have a strong shape- or size-directing functionality in the reaction process. The field-emission properties of the Cu2S nanowires in a vacuum were studied by the Folwer–Nordheim (F–N) theory. The Cu2S nanowires have a low turn-on field at 1.19 V/μm and a high enhancement factor (β) of 19,381. The photocatalytic degradation of Cu2S nanowires was investigated by the change in the concentrations of rhodamine B (RhB) under UV illumination. These outstanding results of Cu2S nanowires indicate that they will be developed as good candidates as electron field emitters and chemical photocatalysts in future nanoelectronic devices.
Vitamin C for Cardiac Surgery Patients: Several Errors in a Published Meta-Analysis. Comment on “Effects of Vitamin C on Organ Function in Cardiac Surgery Patients: A Systematic Review and Meta-Analysis. Nutrients 2019, 11, 2103”
Authors: Hemilä Harri , Chalker Elizabeth
Published: February 2020

We recently published a meta-analysis on vitamin C and the length of intensive care unit [ICU] stay [...]
Positive Association of Dietary Inflammatory Index with Incidence of Cardiovascular Disease: Findings from a Korean Population-Based Prospective Study
Authors: Khan Imran , Kwon Minji , Shivappa Nitin , Hébert James R. , Kim Mi Kyung
Published: February 2020

Recently, diets with higher inflammatory potentials based on the dietary inflammatory index (DII®) have been shown to be associated with increased cardiovascular disease (CVD) risk in the general population. We aimed to prospectively investigate the association between the DII and CVD risk in the large Korean Genome and Epidemiology Study_Health Examination (KoGES_HEXA) cohort comprised of 162,773 participants (men 55,070; women 107,703). A validated semi-quantitative food frequency questionnaire (SQ-FFQ) was used to calculate the DII score. Statistical analyses were performed by using a multivariable Cox proportional hazard model. During the mean follow-up of 7.4 years, 1111 cases of CVD were diagnosed. Higher DII score was associated with increased risk of CVD in men (hazard ratio [HR]Quintile 5 vs. 1 1.43; 95% CI 1.04–1.96) and in women (HRQuintile 5 vs. 1 1.19; 95% CI 0.85–1.67), although not significant for women. The risk of CVD was significantly higher in physically inactive men (HRQuintile 5 vs. 1 1.80; 95% CI 1.03–3.12), obese men (HRQuintile 5 vs. 1 1.77; 95% CI 1.13–2.76) and men who smoked (HRQuintile 5 vs. 1 1.60; 95% CI 1.10–2.33), respectively. The risk of developing stroke was significantly higher for men (HRQuintile 5 vs. 1 2.06; 95% CI 1.07–3.98; p = 0.003), but not for women. A pro-inflammatory diet, as indicated by higher DII scores, was associated with increased risk of CVD and stroke among men.
Supplementation of Mother’s Own Milk with Donor Milk in Infants with Gastroschisis or Intestinal Atresia: A Retrospective Study
Authors: Hoban Rebecca , Khatri Supriya , Patel Aloka , Unger Sharon L.
Published: February 2020

Background: Mother’s own milk (MOM) improves in-hospital outcomes for preterm infants. If unavailable, donor milk (DM) is often substituted. It is unclear if DM vs. formula to supplement MOM is associated with improved in-hospital outcomes in term/late preterm surgical infants with gastroschisis or intestinal atresia. Methods: This retrospective study included infants born ≥33 weeks gestational age (GA) with a birth weight of >1500 g who were admitted to a quaternary neonatal intensive care unit (NICU). Using Chi square and Mann-Whitney u testing, we compared hospital outcomes (length of stay, parenteral nutrition and central line days) before and after a clinical practice change to offer DM instead of formula in this surgical population. Results: Baseline characteristics were similar between eras for the 140 infants (median GA 37 weeks). Fewer infants in DM era were receiving formula at discharge (50.0% vs. 31.4%, p = 0.03). In sub-analyses including only small bowel atresia and gastroschisis infants, the median length of stay (35 vs. 25, p < 0.01) and the central line days (28 vs. 20, p < 0.01) were lower in the DM era. Conclusion: In this retrospective study, offering DM instead of formula was associated with less formula feeding at discharge, and in infants with gastroschisis or small bowel atresia, shorter length of stay and central line days.
Metabolism of cis- and trans-Resveratrol and Dihydroresveratrol in an Intestinal Epithelial Model
Authors: Jarosova Veronika , Vesely Ondrej , Doskocil Ivo , Tomisova Katerina , Marsik Petr , Jaimes Jose D. , Smejkal Karel , Kloucek Pavel , Havlik Jaroslav
Published: February 2020

Trans-resveratrol, a well-known plant phenolic compound, has been intensively investigated due to its association with the so-called French paradox. However, despite its high pharmacological potential, trans-resveratrol has shown relatively low bioavailability. Trans-resveratrol is intensively metabolized in the intestine and liver, yielding metabolites that may be responsible for its high bioactivity. The aim of this study was to investigate and compare the metabolism of trans-resveratrol (tRes), cis-resveratrol (cRes) and dihydroresveratrol (dhRes) in an in vitro epithelial model using Caco-2 cell lines. Obtained metabolites of tRes, cRes and dhRes were analyzed by LC/MS Q-TOF, and significant differences in the metabolism of each compound were observed. The majority of tRes was transported unchanged through the Caco-2 cells, while cRes was mostly metabolized. The main metabolite of both cis- and trans-resveratrol observed as a result of colon microbial metabolism, dhRes, was metabolized almost completely, with only traces of the unchanged molecule being found. A sulphate conjugate was identified as the main metabolite of tRes in our model, while a glucuronide conjugate was the major metabolite of cRes and dhRes. Since metabolism of simple phenolics and polyphenols plays a crucial role in their bioavailability, detailed knowledge of their transformation is of high scientific value.
Gut Microbiota as Important Mediator Between Diet and DNA Methylation and Histone Modifications in the Host
Authors: D’Aquila Patrizia , Lynn Carelli Laurie , De Rango Francesco , Passarino Giuseppe , Bellizzi Dina
Published: February 2020

The human gut microbiota is a complex ecosystem consisting of trillions of microorganisms that inhabit symbiotically on and in the human intestine. They carry out, through the production of a series of metabolites, many important metabolic functions that complement the activity of mammalian enzymes and play an essential role in host digestion. Interindividual variability of microbiota structure, and consequently of the expression of its genes (microbiome), was largely ascribed to the nutritional regime. Diet influences microbiota composition and function with short- and long-term effects. In spite of the vast literature, molecular mechanisms underlying these effects still remain elusive. In this review, we summarized the current evidence on the role exerted by gut microbiota and, more specifically, by its metabolites in the establishment of the host epigenome. The interest in this topic stems from the fact that, by modulating DNA methylation and histone modifications, the gut microbiota does affect the cell activities of the hosting organism.
Australian and New Zealand Medical Students’ Attitudes and Confidence towards Providing Nutrition Care in Practice
Authors: Lepre Breanna , Crowley Jennifer , Mpe Dineo , Bhoopatkar Harsh , Mansfield Kylie J. , Wall Clare , Beck Eleanor J.
Published: February 2020

The prevalence of lifestyle-related chronic disease is increasing. Doctors in primary care are ideally placed to support patient nutrition care, but recent reviews show education is still lacking. This study aimed to identify medical students’ attitudes towards the role of nutrition in health, nutrition knowledge, and perceptions of nutrition education, in postgraduate (Australia) and undergraduate (New Zealand) programs in order to identify gaps in nutrition knowledge and skills to better inform future education. Second-year graduate and third-year undergraduate students participated in semi-structured focus groups and interviews. A general inductive approach was used to investigate students’ (1) attitudes toward the role of nutrition in health, (2) nutrition knowledge based on nutrition-specific competencies and (3) perceived adequacy of nutrition education received. Interviews (nine) and focus groups (seven) identified four common themes: (1) role of medical practitioners in nutrition care, (2) barriers to nutrition education, (3) nutrition knowledge, and (4) nutrition-related skills. Students perceive that doctors are well-placed to provide some level of nutrition care, but poor translation of nutrition knowledge to clinical contexts is a key limitation in nutrition education. In summary, nutrition education may be insufficient to support the nutrition-related competency development of the undergraduate and postgraduate student participants in this study. Focusing on the integration of these skills into the curriculum may be a priority.
First Case of Typhoid Fever due to Extensively Drug-resistant Salmonella enterica serovar Typhi in Italy
Authors: Procaccianti Michela , Motta Alice , Giordani Stefano , Riscassi Sara , Guidi Battista , Ruffini Monica , Maffini Valentina , Esposito Susanna , Dodi Icilio
Published: February 2020

Typhoid fever is a potentially severe and occasionally life-threatening bacteraemic illness caused by Salmonella enterica serovar Typhi (S. Typhi). In Pakistan, an outbreak of extensively drug-resistant (XDR) S. Typhi cases began in November 2016. We report on a five-year-old boy who contracted enteric fever while travelling in Pakistan and was diagnosed after returning to Italy in September 2019. Blood culture isolated Salmonella enterica serovar Typhi that was XDR to all first-line antibiotics, including ceftriaxone and fluoroquinolones. Empiric therapy was switched to meropenem, and the patient recovered completely. Whole-genome sequencing showed that this isolate was of haplotype H58. The XDR S. Typhi clone encoded a chromosomally located resistance region and harbored a plasmid encoding additional resistance elements, including the blaCTX-M-15 extended-spectrum β-lactamase and the qnrS fluoroquinolone resistance gene. This is the first case of typhoid fever due to XDR S. Typhi detected in Italy and one of the first paediatric cases reported outside Pakistan, highlighting the need to be vigilant for future cases. While new vaccines against typhoid are in development, clinicians should consider adapting their empiric approach for patients returning from regions at risk of XDR S. Typhi outbreak with typhoid symptoms.
Simultaneous Immunization with Omp25 and L7/L12 Provides Protection against Brucellosis in Mice
Authors: Gupta Sonal , Mohan Surender , Somani Vikas Kumar , Aggarwal Somya , Bhatnagar Rakesh
Published: February 2020

Currently used Brucella vaccines, Brucella abortus strain 19 and RB51, comprises of live attenuated Brucella strains and prevent infection in animals. However, these vaccines pose potential risks to recipient animals such as attenuation reversal and virulence in susceptible hosts on administration. In this context, recombinant subunit vaccines emerge as a safe and competent alternative in combating the disease. In this study, we formulated a divalent recombinant vaccine consisting of Omp25 and L7/L12 of B. abortus and evaluated vaccine potential individually as well as in combination. Sera obtained from divalent vaccine (Omp25+L7/L12) immunized mice group exhibited enhanced IgG titers against both components and indicated specificity upon immunoblotting reiterating its authenticity. Further, the IgG1/IgG2a ratio obtained against each antigen predicted a predominant Th2 immune response in the Omp25+L7/L12 immunized mice group. Upon infection with virulent B. abortus 544, Omp25+L7/L12 infected mice exhibited superior Log10 protection compared to individual vaccines. Consequently, this study recommends that simultaneous immunization of Omp25 and L7/L12 as a divalent vaccine complements and triggers a Th2 mediated immune response in mice competent of providing protection against brucellosis.
Targeted Protein Degradation by Chimeric Compounds using Hydrophobic E3 Ligands and Adamantane Moiety
Authors: Shoda Takuji , Ohoka Nobumichi , Tsuji Genichiro , Fujisato Takuma , Inoue Hideshi , Demizu Yosuke , Naito Mikihiko , Kurihara Masaaki
Published: February 2020

Targeted protein degradation using small chimeric molecules, such as proteolysis-targeting chimeras (PROTACs) and specific and nongenetic inhibitors of apoptosis protein [IAP]-dependent protein erasers (SNIPERs), is a promising technology in drug discovery. We recently developed a novel class of chimeric compounds that recruit the aryl hydrocarbon receptor (AhR) E3 ligase complex and induce the AhR-dependent degradation of target proteins. However, these chimeras contain a hydrophobic AhR E3 ligand, and thus, degrade target proteins even in cells that do not express AhR. In this study, we synthesized new compounds in which the AhR ligands were replaced with a hydrophobic adamantane moiety to investigate the mechanisms of AhR-independent degradation. Our results showed that the compounds, 2, 3, and 16 induced significant degradation of some target proteins in cells that do not express AhR, similar to the chimeras containing AhR ligands. However, in cells expressing AhR, 2, 3, and 16 did not induce the degradation of other target proteins, in contrast with their response to chimeras containing AhR ligands. Overall, it was suggested that target proteins susceptible to the hydrophobic tagging system are degraded by chimeras containing hydrophobic AhR ligands even without AhR.
Topical Amphotericin B Semisolid Dosage Form for Cutaneous Leishmaniasis: Physicochemical Characterization, Ex Vivo Skin Permeation and Biological Activity
Authors: Berenguer Diana , Alcover Maria Magdalena , Sessa Marcella , Halbaut Lyda , Guillén Carme , Boix-Montañés Antoni , Fisa Roser , Calpena-Campmany Ana Cristina , Riera Cristina , Sosa Lilian
Published: February 2020

Amphotericin B (AmB) is a potent antifungal successfully used intravenously to treat visceral leishmaniasis but depending on the Leishmania infecting species, it is not always recommended against cutaneous leishmaniasis (CL). To address the need for alternative topical treatments of CL, the aim of this study was to elaborate and characterize an AmB gel. The physicochemical properties, stability, rheology and in vivo tolerance were assayed. Release and permeation studies were performed on nylon membranes and human skin, respectively. Toxicity was evaluated in macrophage and keratinocyte cell lines, and the activity against promastigotes and intracellular amastigotes of Leishmania infantum was studied. The AmB gel remained stable for a period of two months, with optimal properties for topical use and no apparent toxic effect on the cell lines. High amounts of AmB were found in damaged and non-damaged skin (1230.10 ± 331.52 and 2484.57 ± 439.12 µg/g/cm2, respectively) and they were above the IC50 of AmB for amastigotes. Although there were no differences in the in vitro anti-leishmanial activity between the AmB solution and gel, the formulation resulted in a higher amount of AmB being retained in the skin, and is therefore a candidate for further studies of in vivo efficacy.
Evaluation of Formulation Parameters on Permeation of Ibuprofen from Topical Formulations Using Strat-M® Membrane
Authors: Bolla Pradeep Kumar , Clark Bradley A. , Juluri Abhishek , Cheruvu Hanumanth Srikanth , Renukuntla Jwala
Published: February 2020

Topical drug delivery is an attractive alternative to conventional methods because of advantages such as non-invasive delivery, by-pass of first pass metabolism, and improved patient compliance. However, several factors such as skin, physicochemical properties of the drug, and vehicle characteristics influence the permeation. Within a formulation, critical factors such as concentration of drug, physical state of drug in the formulation, and organoleptic properties affect the flux across the skin. The aim of the study was to develop and investigate topical semisolid preparations (creams and gels) with ibuprofen as the model drug and investigate the effect of various formulation parameters on the in-vitro performance across the Strat-M® membrane using flow-through cells. In addition, the physical stability of the developed formulations was investigated by studying viscosity, pH, and appearance. All the formulations developed in the study had appealing appearance with smooth texture and no signs of separation. Viscosity and pH of the formulations were acceptable. Cumulative amount of drug permeated at the end of 24 h was highest for clear gel (3% w/w ibuprofen; F6: 739.6 ± 36.1 µg/cm2) followed by cream with high concentration of ibuprofen in suspended form (5% w/w; F3: 320.8 ± 17.53 µg/cm2), emulgel (3% w/w ibuprofen; F5: 178.5 ± 34.5 µg/cm2), and cream with solubilized ibuprofen (3% w/w; F2A: 163.2 ± 9.36 µg/cm2). Results from this study showed that permeation of ibuprofen was significantly influenced by formulation parameters such as concentration of ibuprofen (3% vs. 5% w/w), physical state of ibuprofen (solubilized vs. suspended), formulation type (cream vs. gel), mucoadhesive agents, and viscosity (high vs. low). Thus, findings from this study indicate that pharmaceutical formulation scientists should explore these critical factors during the early development of any new topical drug product in order to meet pre-determined quality target product profile.
Alternatives to Biological Skin in Permeation Studies: Current Trends and Possibilities
Authors: Neupane Rabin , Boddu Sai H.S. , Renukuntla Jwala , Babu R. Jayachandra , Tiwari Amit K.
Published: February 2020

The transdermal route of drugs has received increased attention in recent years due to numerous advantages over the oral and injectable routes, such as avoidance of the hepatic metabolism, protection of drugs from the gastrointestinal tract, sustained drug delivery, and good patient compliance. The assessment of ex vivo permeation during the pharmaceutical development process helps in understanding the product quality and performance of a transdermal delivery system. Generally, excised human skin relevant to the application site or animal skin is recommended for ex vivo permeation studies. However, the limited availability of the human skin and ethical issues surrounding the use of animal skin rendered these models less attractive in the permeation study. In the last three decades, enormous efforts have been put into developing artificial membranes and 3D cultured human skin models as surrogates to the human skin. This manuscript provides an insight on the European Medicines Agency (EMA) guidelines for permeation studies and the parameters affected when using Franz diffusion cells in the permeation study. The need and possibilities for skin alternatives, such as artificially cultured human skin models, parallel artificial membrane permeability assays (PAMPA), and artificial membranes for penetration and permeation studies, are comprehensively discussed.
The Biological Potential Hidden in Inclusion Bodies
Authors: Gifre-Renom Laia , Seras-Franzoso Joaquin , Rafael Diana , Andrade Fernanda , Cano-Garrido Olivia , Martinez-Trucharte Francesc , Ugarte-Berzal Estefania , Martens Erik , Boon Lise , Villaverde Antonio , Opdenakker Ghislain , Schwartz Simó , Arís Anna , Garcia-Fruitós Elena
Published: February 2020

Inclusion bodies (IBs) are protein nanoclusters obtained during recombinant protein production processes, and several studies have demonstrated their potential as biomaterials for therapeutic protein delivery. Nevertheless, IBs have been, so far, exclusively sifted by their biological activity in vitro to be considered in further protein-based treatments in vivo. Matrix metalloproteinase-9 (MMP-9) protein, which has an important role facilitating the migration of immune cells, was used as model protein. The MMP-9 IBs were compared with their soluble counterpart and with MMP-9 encapsulated in polymeric-based micelles (PM) through ionic and covalent binding. The soluble MMP-9 and the MMP-9-ionic PM showed the highest activity values in vitro. IBs showed the lowest activity values in vitro, but the specific activity evolution in 50% bovine serum at room temperature proved that they were the most stable format. The data obtained with the use of an air-pouch mouse model showed that MMP-9 IBs presented the highest in vivo activity compared to the soluble MMP-9, which was associated only to a low and a transitory peak of activity. These results demonstrated that the in vivo performance is the addition of many parameters that did not always correlate with the in vitro behavior of the protein of interest, becoming especially relevant at evaluating the potential of IBs as a protein-based nanomaterial for therapeutic purposes.
Improved Pharmacokinetics and Tissue Uptake of Complexed Daidzein in Rats
Authors: Kwiecień Anna , Ruda-Kucerova Jana , Kamiński Kamil , Babinska Zuzana , Popiołek Iwona , Szczubiałka Krzysztof , Nowakowska Maria , Walczak Maria
Published: February 2020

The pharmacokinetic profile and tissue uptake of daidzein (DAI) was determined in rat serum and tissues (lungs, eyes, brain, heart, spleen, fat, liver, kidney, and testes) after intravenous and intraperitoneal administration of DAI in suspension or complexed with ethylenediamine-modified γ-cyclodextrin (GCD-EDA/DAI). The absolute and relative bioavailability of DAI suspended (20 mg/kg i.v. vs. 50 mg/kg i.p.) and complexed (0.54 mg/kg i.v. vs. 1.35 mg/kg i.p.) was determined. After i.p. administration, absorption of DAI complexed with GCD-EDA was more rapid (tmax = 15 min) than that of DAI in suspension (tmax = 45 min) with a ca. 3.6 times higher maximum concentration (Cmax = 615 vs. 173 ng/mL). The i.v. half-life of DAI was longer in GCD-EDA/DAI complex compared with DAI in suspension (t0.5 = 380 min vs. 230 min). The volume of distribution of DAI given i.v. in GCD-EDA/DAI complex was ca. 6 times larger than DAI in suspension (38.6 L/kg vs. 6.2 L/kg). Our data support the concept that the pharmacokinetics of DAI suspended in high doses are nonlinear. Increasing the intravenous dose 34 times resulted in a 5-fold increase in AUC. In turn, increasing the intraperitoneal dose 37 times resulted in a ca. 2-fold increase in AUC. The results of this study suggested that GCD-EDA complex may improve DAI bioavailability after i.p. administration. The absolute bioavailability of DAI in GCD-EDA inclusion complex was ca. 3 times greater (F = 82.4% vs. 28.2%), and the relative bioavailability was ca. 21 times higher than that of DAI in suspension, indicating the need to study DAI bioavailability after administration by routes other than intraperitoneal, e.g., orally, subcutaneously, or intramuscularly. The concentration of DAI released from GCD-EDA/DAI inclusion complex to all the rat tissues studied was higher than after administration of DAI in suspension. The concentration of DAI in brain and lungs was found to be almost 90 and 45 times higher, respectively, when administered in complex compared to the suspended DAI. Given the nonlinear relationship between DAI bioavailability and the dose released from the GCD-EDA complex, complexation of DAI may thus offer an effective approach to improve DAI delivery for treatment purposes, for example in mucopolysaccharidosis (MPS), allowing the reduction of ingested DAI doses.
Improved Biosafety and Transdermal Delivery of Aconitine via Diethylene Glycol Monoethyl Ether-Mediated Microemulsion Assisted with Microneedles
Authors: Zhang Yongtai , Hu Hongmei , Jing Qian , Wang Zhi , He Zehui , Wu Tong , Feng Nian-Ping
Published: February 2020

In the current study, diethylene glycol monoethyl ether-mediated microemulsions were combined with microneedles for enhanced transdermal aconitine delivery. The oil-in-water microemulsion increasedaconitine solubility and enhanced transdermal drug delivery and assistance with metal microneedles enhanced permeation of the aconitine-loaded microemulsion. Carried by the microemulsion, the in vitro permeability of aconitine was significantly enhanced, and further improved using microneedles. In vivo microdialysis revealed that the subcutaneous local drug concentration reached a high level within 30 min and remained relatively consistent to the end of the experimental period. AUC0-t of the microemulsion group was significantly higher than that of the aqueous solution group, and the microemulsion combined with microneedles group achieved the highest AUC0-t among the tested groups. The microemulsion and microdialysis probe also showed good biocompatibility with skin tissue. The microemulsion could be internalized by HaCaT and CCC-ESF-1 cells via lysosomes. The in vitro cytotoxicity of aconitine toward skin cells was reduced via encapsulation by microemulsion, and the prepared microemulsion developed no skin irritation. Hence, transdermal aconitine delivery and drug biosafety were effectively improved by loading into the microemulsion and assisting with microneedles, and in vivo microdialysis technique is suitable for realtime monitoring of transdermal drug delivery with microemulsion-based drug vehicles.
A Mixed Micellar Formulation for the Transdermal Delivery of an Indirubin Analog
Authors: Seo Seol Hwa , Kim Eunhwan , Joo Yechan , Lee Juseung , Oh Kyung Taek , Hwang Sung-Joo , Choi Kang-Yell
Published: February 2020

Indirubin is an active component of Dang Gui Long Hui Wan, which has been used in traditional Chinese medicine to treat inflammatory diseases as well as for the prevention and treatment of human cancer, such as chronic myeloid leukemia. The therapeutic effects of indirubin analogs have been underestimated due to its poor water solubility and low bioavailability. To improve the solubility and bioavailability of indirubin analogs, we prepared a mixed micellar formulation with Kolliphor® EL and Tween 80 as surfactants, and PEG 400 as a co-surfactant, followed by complexation with (2-hydroxyproply)-β-cyclodextrin at appropriate ratios. Overall, improving the solubility and skin penetration of indirubin analogs can increase clinical efficacy and provide maximum flux through the skin.
Non-Viral in Vitro Gene Delivery: It is Now Time to Set the Bar!
Authors: Bono Nina , Ponti Federica , Mantovani Diego , Candiani Gabriele
Published: February 2020

Transfection by means of non-viral gene delivery vectors is the cornerstone of modern gene delivery. Despite the resources poured into the development of ever more effective transfectants, improvement is still slow and limited. Of note, the performance of any gene delivery vector in vitro is strictly dependent on several experimental conditions specific to each laboratory. The lack of standard tests has thus largely contributed to the flood of inconsistent data underpinning the reproducibility crisis. A way researchers seek to address this issue is by gauging the effectiveness of newly synthesized gene delivery vectors with respect to benchmarks of seemingly well-known behavior. However, the performance of such reference molecules is also affected by the testing conditions. This survey points to non-standardized transfection settings and limited information on variables deemed relevant in this context as the major cause of such misalignments. This review provides a catalog of conditions optimized for the gold standard and internal reference, 25 kDa polyethyleneimine, that can be profitably replicated across studies for the sake of comparison. Overall, we wish to pave the way for the implementation of standardized protocols in order to make the evaluation of the effectiveness of transfectants as unbiased as possible.
MiR-219a-5p Enriched Extracellular Vesicles Induce OPC Differentiation and EAE Improvement More Efficiently Than Liposomes and Polymeric Nanoparticles
Authors: Osorio-Querejeta Iñaki , Carregal-Romero Susana , Ayerdi-Izquierdo Ana , Mäger Imre , Nash Leslie A. , Wood Matthew , Egimendia Ander , Betanzos M. , Alberro Ainhoa , Iparraguirre Leire , Moles Laura , Llarena Irantzu , Möller Marco , Goñi-de-Cerio Felipe , Bijelic Goran , Ramos-Cabrer Pedro , Muñoz-Culla Maider , Otaegui David
Published: February 2020

Remyelination is a key aspect in multiple sclerosis pathology and a special effort is being made to promote it. However, there is still no available treatment to regenerate myelin and several strategies are being scrutinized. Myelination is naturally performed by oligodendrocytes and microRNAs have been postulated as a promising tool to induce oligodendrocyte precursor cell differentiation and therefore remyelination. Herein, DSPC liposomes and PLGA nanoparticles were studied for miR-219a-5p encapsulation, release and remyelination promotion. In parallel, they were compared with biologically engineered extracellular vesicles overexpressing miR-219a-5p. Interestingly, extracellular vesicles showed the highest oligodendrocyte precursor cell differentiation levels and were more effective than liposomes and polymeric nanoparticles crossing the blood–brain barrier. Finally, extracellular vesicles were able to improve EAE animal model clinical evolution. Our results indicate that the use of extracellular vesicles as miR-219a-5p delivery system can be a feasible and promising strategy to induce remyelination in multiple sclerosis patients.
Formulating a Stable Mannitol Infusion while Maintaining Hyperosmolarity
Authors: Kavanagh Oisín , Hogan Fiona , Murphy Caoimhe , Croker Denise , Walker Gavin
Published: February 2020

Mannitol infusion is commonly used in the treatment of intracranial hypertension following traumatic brain injury. It has long been known to have stability issues, specifically, mannitol recrystallises from solutions greater than 10% w/v in ambient conditions. This can happen at any time, whether on the pharmacy shelf or during a medical procedure. This study describes the stability limits of 20% w/v mannitol infusion (the most common strength used clinically) and proposes a number of safer, stable and tuneable hyperosmotic formulations of mannitol in combination with clinically acceptable osmotic agents (NaCl, sorbitol and glycerol).
Co-Encapsulation of Mitoxantrone and β-Elemene in Solid Lipid Nanoparticles to Overcome Multidrug Resistance in Leukemia
Authors: Amerigos Daddy J.C. Kambere , Chen Minglei , Raza Faisal , Xiao Yanyu , Su Zhigui , Ping Qineng
Published: February 2020

Multidrug resistance (MDR) due to P-glycoprotein (P-gp) overexpression is a major obstacle to successful leukemia chemotherapy. The combination of anticancer chemotherapy with a chemosensitizer of P-gp inhibitor is promising to overcome MDR, generate synergistic effects, and maximize the treatment effect. Herein, we co-encapsulated a chemotherapeutic drug of mitoxantrone (MTO) and a P-gp inhibitor of β-elemene (βE) in solid lipid nanoparticles (MTO/βE-SLNs) for reversing MDR in leukemia. The MTO/βE-SLNs with about 120 nm particle size possessed good colloidal stability and sustained release behavior. For the cellular uptake study, doxorubicin (DOX) was used as a fluorescence probe to construct SLNs. The results revealed that MTO/βE-SLNs could be effectively internalized by both K562/DOX and K562 cells through the pathway of caveolate-mediated endocytosis. Under the optimized combination ratio of MTO and βE, the in vitro cytotoxicity study indicated that MTO/βE-SLNs showed a better antitumor efficacy in both K562/DOX and K562 cells than other MTO formulations. The enhanced cytotoxicity of MTO/βE-SLNs was due to the increased cellular uptake and blockage of intracellular ATP production and P-gp efflux by βE. More importantly, the in vivo studies revealed that MTO/βE-SLNs could significantly prolong the circulation time and increase plasma half-life of both MTO and βE, accumulate into tumor and exhibit a much higher anti-leukemia effect with MDR than other MTO formulations. These findings suggest MTO/βE-SLNs as a potential combined therapeutic strategy for overcoming MDR in leukemia.
Breaking Barriers: Bioinspired Strategies for Targeted Neuronal Delivery to the Central Nervous System
Authors: Spencer Ana P. , Torrado Marília , Custódio Beatriz , Silva-Reis Sara C. , Santos Sofia D. , Leiro Victoria , Pêgo Ana P.
Published: February 2020

Central nervous system (CNS) disorders encompass a vast spectrum of pathological conditions and represent a growing concern worldwide. Despite the high social and clinical interest in trying to solve these pathologies, there are many challenges to bridge in order to achieve an effective therapy. One of the main obstacles to advancements in this field that has hampered many of the therapeutic strategies proposed to date is the presence of the CNS barriers that restrict the access to the brain. However, adequate brain biodistribution and neuronal cells specific accumulation in the targeted site also represent major hurdles to the attainment of a successful CNS treatment. Over the last few years, nanotechnology has taken a step forward towards the development of therapeutics in neurologic diseases and different approaches have been developed to surpass these obstacles. The versatility of the designed nanocarriers in terms of physical and chemical properties, and the possibility to functionalize them with specific moieties, have resulted in improved neurotargeted delivery profiles. With the concomitant progress in biology research, many of these strategies have been inspired by nature and have taken advantage of physiological processes to achieve brain delivery. Here, the different nanosystems and targeting moieties used to achieve a neuronal delivery reported in the open literature are comprehensively reviewed and critically discussed, with emphasis on the most recent bioinspired advances in the field. Finally, we express our view on the paramount challenges in targeted neuronal delivery that need to be overcome for these promising therapeutics to move from the bench to the bedside.
Effect of Oxaliplatin-Loaded Poly (d,l-Lactide-co-Glycolic Acid) (PLGA) Nanoparticles Combined with Retinoic Acid and Cholesterol on Apoptosis, Drug Resistance, and Metastasis Factors of Colorectal Cancer
Authors: C. de S. L. Oliveira Ana Luiza , Araújo Júnior Raimundo Fernandes de , Gomes de Carvalho Thaís , B. Chan Alan , Schomann Timo , Tamburini Filippo , de Geus-Oei Lioe-Fee , J. Cruz Luis
Published: February 2020

Apoptosis signaling pathways, drug resistance, and metastasis are important targets to develop new cancer treatments. We developed cholesterol-coated Poly(d,l-Lactide-co-Glycolic Acid) (PLGA) nanoparticles for effective encapsulation and delivery of retinoic acid and oxaliplatin to analyze their antitumor activity in colorectal cancer. The cell viability and proliferation of tumoral cells lines (CT-26 and SW-480) decreased when compared to control in vitro after treatment with the nanoparticles. In addition, apoptosis of CT-26 cells increased. Importantly, cytoprotection of nontumor cells was detected. Expression of pro-apoptotic proteins was upregulated, while anti-apoptotic proteins were downregulated either in vitro or in vivo. In addition, drug resistance and metastasis factors were downregulated in vivo. Human colorectal tumors that highly expressed BCL-2 and Ki-67 had a greater tendency towards death within 60 months. Our results show that loading oxaliplatin combined with retinoic acid and cholesterol in a nanoparticle formulation enables determination of optimal antitumor activity and subsequent treatment efficacy.
Stability Study of Isoniazid and Rifampicin Oral Solutions Using Hydroxypropyl-Β-Cyclodextrin to Treat Tuberculosis in Paediatrics
Authors: Santoveña-Estévez Ana , Suárez-González Javier , Cáceres-Pérez Amor R. , Ruiz-Noda Zuleima , Machado-Rodríguez Sara , Echezarreta Magdalena , Soriano Mabel , Fariña José B.
Published: February 2020

(1) Background: First-line antituberculosis treatment in paediatrics entails the administration of Isoniazid, Pyrazinamide, and Rifampicin. This study examines the possibility of developing a combined dose liquid formulation for oral use that would facilitate dose adjustment and adherence to treatment for younger children. (2) Methods: The active pharmaceutical ingredients stability under in vitro paediatric digestive pH conditions have been checked. The samples were studied as individual or fixed combined paediatric dosages to determine the pH of maximum stability. The use of hydroxypropyl-β-cyclodextrin to improve Rifampicin solubility and the use of ascorbic acid to increase the stability of the formulation have been studied. (3) Results: Maximum stability of combined doses was determined at pH 7.4, and maximum complexation at pH 8.0. Taking this into account, formulations presented the minimum dose of two active pharmaceutical ingredients dissolved. The addition of ascorbic acid at 0.1% w/v enables the detection of a higher remaining quantity of both drugs after three days of storage at 5 °C. (4) Conclusions: a formulation which combines the minimum paediatric dosages dissolved recommended by WHO for Isoniazid and Rifampicin has been developed. Future assays are needed to prolong the stability of the formulation with the aim of incorporating Pyrazinamide to the solution.
Physiological and Anatomical Mechanisms in Wheat to Cope with Salt Stress Induced by Seawater
Authors: Nassar Rania M. A. , Kamel Hedaya A. , Ghoniem Ahmed E. , Alarcón Juan José , Sekara Agnieszka , Ulrichs Christian , Abdelhamid Magdi T.
Published: February 2020

Two pot experiments were conducted in a greenhouse to examine 14C fixation and its distribution in biochemical leaf components, as well as the physiological and anatomical adaptability responses of wheat (Triticum aestivum L.) grown with seawater diluted to 0.2, 3.0, 6.0, and 12.0 dS m−1. The results showed significant reductions in chlorophyll content, 14C fixation (photosynthesis), plant height, main stem diameter, total leaf area per plant, and total dry weight at 3.0, 6.0, and 12.0 dS m−1 seawater salt stress. The 14C loss was very high at 12.0 ds m−1 after 120 h. 14C in lipids (ether extract) showed significant changes at 12.0 dS m−1 at 96 and 120 h. The findings indicated the leaf and stem anatomical feature change of wheat plants resulting from adaptation to salinity stress. A reduction in the anatomical traits of stem and leaf diameter, wall thickness, diameter of the hollow pith cavity, total number of vascular bundles, number of large and small vascular bundles, bundle length and width, thickness of phloem tissue, and diameter of the metaxylem vessel of wheat plants was found. In conclusion, salt stress induces both anatomical and physiological changes in the stem and leaf cells of wheat, as well as the tissues and organs, and these changes in turn make it possible for the plants to adapt successfully to a saline environment.
Structure, Function, Diversity, and Composition of Fungal Communities in Rhizospheric Soil of Coptis chinensis Franch under a Successive Cropping System
Authors: Alami Mohammad Murtaza , Xue Jinqi , Ma Yutao , Zhu Dengyan , Abbas Aqleem , Gong Zedan , Wang Xuekui
Published: February 2020

Soil types and cropping systems influence the diversity and composition of the rhizospheric microbial communities. Coptis chinensis Franch is one of the most important medicinal plants in China. In the current study, we provide detailed information regarding the diversity and composition of rhizospheric fungal communities of the C. chinensis plants in continuous cropping fields and fallow fields in two seasons (winter and summer), using next-generation sequencing. Alpha diversity was higher in the five-year C. chinensis field and lower in fallow fields. Significant differences analysis confirmed more fungi in the cultivated field soil than in fallow fields. Additionally, PCoA of beta diversity indices revealed that samples associated with the cultivated fields and fallow fields in different seasons were separated. Five fungal phyla (Ascomycota, Basidiomycota, Chytridiomycota, Glomeromycota and Mucoromycota) were identified from the soil samples in addition to the unclassified fungal taxa and Cryptomycota, and among these phyla, Ascomycota was predominantly found. FUNGuild fungal functional prediction revealed that saprotroph was the dominant trophic type in all two time-series soil samples. Redundancy analysis (RDA) of the dominant phyla data and soil physiochemical properties revealed the variations in fungal community structure in the soil samples. Knowledge from the present study could provide a valuable reference for solving the continuous cropping problems and promote the sustainable development of the C. chinensis industry.
Comparative Analysis of In Vitro Responses and Regeneration between Diverse Bioenergy Sorghum Genotypes
Authors: Flinn Barry , Dale Savanah , Disharoon Andrew , Kresovich Stephen
Published: February 2020

Sorghum has been considered a recalcitrant plant in vitro and suffers from a lack of regeneration protocols that function broadly and efficiently across a range of genotypes. This study was initiated to identify differential genotype-in vitro protocol responses across a range of bioenergy sorghum parental lines and the common grain sorghum genotype Tx430 in order to characterize response profiles for use in future genetic studies. Two different in vitro protocols, LG and WU, were used for comparisons. Distinct genotype-protocol responses were observed, and the WU protocol performed significantly better for plantlet regeneration. Most bioenergy genotypes performed as well, if not better than Tx430, with Rio and PI329311 as the top regenerating lines. Genotypes displayed protocol-dependent, differential phenolic exudation responses, as indicated by medium browning. During the callus induction phase, genotypes prone to medium browning exhibited a response on WU medium which was either equal or greater than on LG medium. Genotype- and protocol-dependent albino plantlet regeneration was also noted, with three of the bioenergy genotypes showing albino plantlet regeneration. Grassl, Rio and Pink Kafir were susceptible to albino plantlet regeneration, with the response strongly associated with the WU protocol. These bioenergy parental genotypes, and their differential responses under two in vitro protocols, provide tools to further explore and assess the role of genetic loci, candidate genes, and allelic variants in the regulation of in vitro responsiveness in sorghum.