Investigation of antimicrobial activity of photothermal therapeutic gold/copper sulfide core/shell nanoparticles to bacterial spores and cells

Background

Au/CuS core/shell nanoparticles (NPs) were designed as a new type of transducer agent for photothermal therapy (PTT), with attractive features of easy preparation, low cost and small size for targeting. This paper studied for the first time the intrinsic antimicrobial activity of Au/CuS NPs to B. anthracis spores and cells in addition to its PTT effect.

Results

It was found that Au/CuS NPs were highly efficient in inactivating B. anthracis cells, but not effective to the spores. Treatment with NPs at ~0.83 μM for 30 min achieved a 7 log reduction in viable cells. The antimicrobial effect was both NPs concentration and treatment time dependent. SEM imaging and the efflux of DNA test demonstrated the damage of cell membrane after NPs treatment, yet further research is necessary to fully understand the precise inactivation mechanism.

Conclusions

The Au/CuS NPs had strong antimicrobial activity to B. anthracis cells, which showed a great potential to be an effective antimicrobial agent to bacterial cells.

     

Biosynthesis of flower-shaped Au nanoclusters with EGCG and their application for drug delivery

Background

In the last decade, the biosynthesis of metal nanoparticles using organisms have received more and more considerations. However, the complex composition of organisms adds up to a great barrier for the characterization of biomolecules involved in the synthesis process and their biological mechanisms.

Results

In this research, we biosynthesized a kind of flower-shaped Au nanoclusters (Au NCs) using one definite component—epigallocatechin gallate (EGCG), which was the main biomolecules of green tea polyphenols. Possessing good stability for 6 weeks and a size of 50 nm, the Au NCs might be a successful candidate for drug delivery. Hence, both methotrexate (MTX) and doxorubicin (DOX) were conjugated to the Au NCs through a bridge of cysteine (Cys). The introduction of MTX provided good targeting property for the Au NCs, and the conjugation of DOX provided good synergistic effect. Then, a novel kind of dual-drug loaded, tumor-targeted and highly efficient drug delivery system (Au-Cys-MTX/DOX NCs) for combination therapy was successfully prepared. The TEM of HeLa cells incubated with Au-Cys-MTX/DOX NCs indicated that the Au-Cys-MTX/DOX NCs could indeed enter and kill cancer cells. The Au-Cys-MTX/DOX NCs also possessed good targeting effect to the FA-receptors-overpressed cancer cells both in vitro and in vivo. Importantly, the Au-Cys-MTX/DOX NCs resulted in an excellent anticancer activity in vivo with negligible side effects.

Conclusions

These results suggest that the biosynthesized Au-Cys-MTX/DOX NCs could be a potential carrier with highly efficient anticancer properties for tumor-targeted drug delivery.

     

Characterization of the nonlinear optical properties of nanocrystals by Hyper Rayleigh Scattering

Background

Harmonic Nanoparticles are a new family of exogenous markers for multiphoton imaging exerting optical contrast by second harmonic (SH) generation. In this tutorial, we present the application of Hyper-Rayleigh Scattering (HRS) for a quantitative assessment of the nonlinear optical properties of these particles and discuss the underlying theory and some crucial experimental aspects.

Methods

The second harmonic properties of BaTiO₃, KNbO₃, KiTiOPO₄ (KTP), LiNbO₃ and ZnO nanocrystals (NCs) are investigated by HRS measurements after careful preparation and characterization of colloidal suspensions.

Results

A detailed analysis of the experimental results is presented with emphasis on the theoretical background and on the influence of some experimental parameters including the accurate determination of the nanocrystal size and concentration. The SH generation efficiency and averaged nonlinear optical coefficients are then derived and compared for six different types of NCs.

Conclusions

After preparation of colloidal NC suspensions and careful examination of their size, concentration and possible aggregation state, HRS appears as a valuable tool to quantitatively assess the SH efficiency of noncentrosymmetric NCs. All the investigated nanomaterials show high SH conversion efficiencies, demonstrating a good potential for bio-labelling applications.

     

Endophytic infection modifies organic acid and mineral element accumulation by rice under Na<Subscript>2</Subscript>CO<Subscript>3</Subscript> stress

Background and aims

Saline and alkali soils severely impact plant growth. Endophyte and plant associations are known to significantly modify plant metabolism. This study reports the effects of a type of endophyte on organic acid (OA) accumulation and ionic balance in rice under Na₂CO₃ stress.

Methods

Rice seedlings with (E+) and without (E-) endophytic infection were subjected to different levels of Na₂CO₃ stress (0, 5, 10, 15, and 20 mM) for two weeks. Organic acids and mineral elements in the leaves and roots were determined.

Results

Seedlings with endophytic infection accumulated mainly citrate and fumarate, with some malate and succinate in the leaves. In the roots, accumulation of malate and fumarate was enhanced significantly by endophytic infection, while less citrate and succinate was accumulated under Na₂CO₃ stress, which suggested that leaves and roots use different mechanisms to control OA metabolism. Endophytes reduced the total Na and Na:K ratios, but increased ST values, the percent changes of other measured nutrients, Chl content, and dry weight per plant under Na₂CO₃ stress.

Conclusions

Endophytic infection plays a key role in maintaining plant growth by improving nutrient uptake and adjusting OA accumulation under Na₂CO₃ stress. The application of endophytes can enhance the resistance of rice to salinity.

     

Sublethal concentration of H<Subscript>2</Subscript>O<Subscript>2</Subscript> enhances the protective effect of mesenchymal stem cells in rat model of spinal cord injury

Objective

To investigate the effect of H₂O₂ on the migration and antioxidant defense of mesenchymal stem cells (MSCs) and the neurotrophic effects of H₂O₂-treated MSCs on spinal cord injury (SCI).

Results

Sublethal concentrations of H₂O₂ decreased cell migration and expression of CXCR4 and CCR2 as well as Nrf2 expression in MSCs. In the second phase, transplantation of treated and untreated MSCs to SCI caused minor changes in locomotor dysfunction. There was a significantly difference between cell-treated and spinal cord injury groups in expression of BDNF (brain-derived neurotrophic factor). Transplantation of H₂O₂-treated cells caused an increase in BDNF expression compared to non-treated cells.

Conclusion

Transplantation of H₂O₂-treated stem cells may have protective effects against SCI through by increasing neurotrophic factors.

     

Life cycle energy use and CO<Subscript>2</Subscript> emissions of small-scale gold mining and refining processes in the Philippines

Purpose

Gold is one of the most significant metals in the world, with use in various sectors including the electronic, health, and fashion industries. The Philippines has the world’s third largest known Au deposits and is ranked 20th in global gold production. Of the country’s annual production, about 80% is from the small-scale gold mining (SSGM) sector. This work estimates the first location-specific life cycle energy use and CO₂ emissions of SSGM establishments in the Philippines.

Methods

Process-based LCA was used with functional unit of 100 g Au and observed data from 2010 to 2011 for mining, comminution, recovery, and refining. Four gold production paths were observed in the provinces of Benguet and Camarines Norte, namely, amalgamation, cyanidation with carbon-in-leach (CIL), cyanidation with leaching with zinc, and combination of amalgamation and cyanidation with CIL.

Results and discussion

It was estimated that 3–18 g of Au was extracted for every ton of ore within 57–159 man-hours from mining to refining. Energy use estimates ranged from 3501 to 67,325 MJ/100 g Au, while CO₂ emission estimates ranged from 398 to 5340 kg CO₂/100 g Au. The combination of amalgamation and cyanidation with CIL processes was the least energy and carbon intensive, while cyanidation with CIL process was the most intensive. Electricity use accounted for 95–100% of total emissions, except in cyanidation with CIL where kerosene accounts for 77% of the total. Since SSGMs contributed 80% of the 40 tons of Au produced in the Philippines in 2014, the SSGM energy use was estimated to be between 1120 and 21,544 TJ and the CO₂ emissions to be between 129 and 1726 ktons CO₂. Energy estimates are most sensitive to refining process yield and electrical equipment efficiency.

Conclusions

The estimated life cycle emissions rate for SSGM in the Philippines is lower than available estimates of large-scale mining. Notwithstanding, given the sector’s reliance on fossil fuels for its energy needs and the Philippines’ pledge to reduce its CO₂ footprint by 70% in 2030, every effort to mitigate energy use and CO₂ emission counts. Three main recommendations toward energy consumption and CO₂ emissions reduction in SSGMs are proposed: (1) policy to promote technologies that are energy-efficient and processes that maximize gold process yield, (2) effective Minahang Bayan (SSGM mining zone mandated by law) implementation to ensure use of higher-grade ores, and (3) adoption of renewable energy in Minahang Bayans to promote energy independence and mitigate CO₂ emissions.

     

The effect of 1,25-dihydroxyvitamin D<Subscript>3</Subscript> on TSLP,IL-33 and IL-25 expression in respiratory epithelium

Background

Airway epithelium is an active and important component of the immunological response in the pathophysiology of obstructive lung diseases. Recent studies suggest an important role for vitamin D₃ in asthma severity and treatment response.

Objective

Our study evaluated the influence of an active form of vitamin D₃ on the expression of selected mediators of allergic inflammation in the respiratory epithelium.

Material and Methods

Primary nasal and bronchial epithelial cells were exposed to1,25D₃ for 1 hour and were then stimulated or not with IL-4, TNF-α, LPS, and poly I:C. After 24 hours TSLP, IL-33, and IL-25 protein levels were measured in culture supernatants usingELISAandmRNAlevels in cells by real time PCR.

Results

1,25D₃ increased TSLP concentration in unstimulated nasal epithelial cells, but did not influence IL-33 and IL-25 expression. In IL-4-stimulated epithelial cell cultures 1,25D₃ mostly inhibited TSLP and IL-33 expression. In LPS-treated cultures 1,25D₃ decreased IL-33 expression. Simultaneously 1,25D₃ augmented IL-25 production in the same model of stimulation.

Conclusion

Our study revealed the dual nature of vitamin D₃ manifested in both pro- and anti-inflammatory properties observed in airway epithelial cells.

     

Effectiveness of fractional CO<Subscript>2</Subscript> laser in women with stress urinary incontinence

Background

Stress urinary incontinence (SUI) is a relatively common disorder that significantly affects the quality of life. Many conservative and surgical treatment methods have been recommended for SUI, but they have major limitations.

Aims

To assess the use of the CO₂ fractional laser in the treatment of SUI.

Methods

This clinical trial included 55 patients with confirmed SUI. Patients underwent fractional CO₂ laser treatment 3 times at 30-day intervals. Data on age, smoking history, sexual activity, menopause, and history of hormone replacement therapy (HRT) were collected. Response to treatment was assessed by SUI severity and the level of sexual satisfaction was assessed using the visual analog scale (VAS). Patients were evaluated at 3 different time points: before treatment, and 45 days and 6 months after the last laser treatment.

Results

The mean patient age was 44.4±11.4 years (range: 28 to 68 years). Smoking history was positive in 6 patients (9.1%); 19 (54.3%) were menopausal on HRT. The SUI severity score at baseline (before treatment) was 8.56±0.62 and decreased to 2.28 6 months after treatment (p<0.0001). The sexual satisfaction score was 3±0.94 at baseline and increased to 7.87±0.93 6 months after treatment (day 180) (p<0.0001, slope = + 2.2)

Conclusion

Our findings are in line with a previous study that showed the value of transvaginal CO₂ fractional laser treatment for alleviation of SUI symptoms and its potential as an alternative treatment. We also observed improved sexual satisfaction in SUI patients.

     

Versatility of chitosan/BmNPV bacmid DNA nanocomplex as transfection reagent of recombinant protein expression in silkworm larvae

Objective

To examine the feasibility of chitosan as an alternative transfection reagent candidate for protein expression in Bm5 cells and silkworm larvae using recombinant BmNPV bacmid DNA.

Result

Chitosan 100 and recombinant Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid DNA, in amino group/phosphate group (N/P) ratios of 0.1–10, were used for formation of chitosan/DNA nanocomplexes. The chitosan/BmNPV bacmid DNA nanocomplexes showed higher specific activity of GFP_uv-β1,3-N-acetylglucosaminyltransferase 2 (β3GnT2) fusion protein (GGT2) expressed in silkworm larvae than DMRIE-C, a conventional silkworm transfection reagent. In particular, the composition of chitosan and BmNPV bacmid DNA nanocomplexes formed by an N/P ratio of 8 or 10, respectively, showed the highest specific activity of β3GnT2 in the silkworm larvae hemolymph. In addition, three different proteins were expressed in silkworm larvae to the same extent using chitosan as that using DMRIE-C.

Conclusion

This is the first finding that chitosan/BmNPV bacmid DNA nanocomplexes can rival the performance of commercially available transfection reagents for the expression of recombinant proteins in Bm5 cells and silkworm larvae.

     

Structures of EccB<Subscript>1</Subscript> and EccD<Subscript>1</Subscript> from the core complex of the mycobacterial ESX-1 type VII secretion system

Background

The ESX-1 type VII secretion system is an important determinant of virulence in pathogenic mycobacteria, including Mycobacterium tuberculosis. This complicated molecular machine secretes folded proteins through the mycobacterial cell envelope to subvert the host immune response. Despite its important role in disease very little is known about the molecular architecture of the ESX-1 secretion system.

Results

This study characterizes the structures of the soluble domains of two conserved core ESX-1 components – EccB₁ and EccD₁. The periplasmic domain of EccB₁ consists of 4 repeat domains and a central domain, which together form a quasi 2-fold symmetrical structure. The repeat domains of EccB₁ are structurally similar to a known peptidoglycan binding protein suggesting a role in anchoring the ESX-1 system within the periplasmic space. The cytoplasmic domain of EccD₁has a ubiquitin-like fold and forms a dimer with a negatively charged groove.

Conclusions

These structures represent a major step towards resolving the molecular architecture of the entire ESX-1 assembly and may contribute to ESX-1 targeted tuberculosis intervention strategies.

     

Human health characterization factors of nano-TiO<Subscript>2</Subscript> for indoor and outdoor environments

Purpose

The increasing use of engineered nanomaterials (ENMs) in industrial applications and consumer products is leading to an inevitable release of these materials into the environment. This makes it necessary to assess the potential risks that these new materials pose to human health and the environment. Life cycle assessment (LCA) methodology has been recognized as a key tool for assessing the environmental performance of nanoproducts. Until now, the impacts of ENMs could not be included in LCA studies due to a lack of characterization factors (CFs). This paper provides a methodological framework for identifying human health CFs for ENMs.

Methods

The USEtox? model was used to identify CFs for assessing the potential carcinogenic and non-carcinogenic effects on human health caused by ENM emissions in both indoor (occupational settings) and outdoor environments. Nano-titanium dioxide (nano-TiO₂) was selected for defining the CFs in this study, as it is one of the most commonly used ENMs. For the carcinogenic effect assessment, a conservative approach was adopted; indeed, a critical dose estimate for pulmonary inflammation was assumed.

Results and discussion

We propose CFs for nano-TiO₂ from 5.5E?09 to 1.43E?02 cases/kg_emitted for both indoor and outdoor environments and for carcinogenic and non-carcinogenic effects.

Conclusions

These human health CFs for nano-TiO₂ are an important step toward the comprehensive application of LCA methodology in the field of nanomaterial technology.

     

Elevated CO<Subscript>2</Subscript> and virus infection impacts wheat and aphid metabolism

Introduction

The aphid Rhopalosiphum padi L. is a vector of Barley yellow dwarf virus (BYDV) in wheat and other economically important cereal crops. Increased atmospheric CO₂ has been shown to alter plant growth and metabolism, enhancing BYDV disease in wheat. However, the biochemical influences on aphid metabolism are not known.

Objectives

This work aims to determine whether altered host-plant quality, influenced by virus infection and elevated CO₂, impacts aphid weight and metabolism.

Methods

Untargeted ¹H NMR metabolomics coupled with multivariate statistics were employed to profile the metabolism of R. padi reared on virus-infected and non-infected (sham-inoculated) wheat grown under ambient CO₂ (aCO₂, 400 µmol mol^?1) and future, predicted elevated CO₂ (eCO₂, 650 µmol mol^?1) concentrations. Un-colonised wheat was also profiled to observe changes to host-plant quality (i.e., amino acids and sugars).

Results

The direct impacts of virus or eCO₂ were compared. Virus presence increased aphid weight under aCO₂ but decreased weight under eCO₂; whilst eCO₂ increased non-viruliferous (sham) aphid weight but decreased viruliferous aphid weight. Discriminatory metabolites due to eCO₂ were succinate and sucrose (in sham wheat), glucose, choline and betaine (in infected wheat), and threonine, lactate, alanine, GABA, glutamine, glutamate and asparagine (in aphids), irrespective of virus presence. Discriminatory metabolites due to virus presence were alanine, GABA, succinate and betaine (in wheat) and threonine and lactate (in aphids), irrespective of CO₂ treatment.

Conclusion

This study confirms that virus and eCO₂ alter host-plant quality, and these differences are reflected by aphid weight and metabolism.

     

Mitofusin 1 is degraded at G<Subscript>2</Subscript>/M phase through ubiquitylation by MARCH5

Background

Mitochondria exhibit a dynamic morphology in cells and their biogenesis and function are integrated with the nuclear cell cycle. In mitotic cells, the filamentous network structure of mitochondria takes on a fragmented form. To date, however, whether mitochondrial fusion activity is regulated in mitosis has yet to be elucidated.

Findings

Here, we report that mitochondria were found to be fragmented in G₂ phase prior to mitotic entry. Mitofusin 1 (Mfn1), a mitochondrial fusion protein, interacted with cyclin B1, and their interactions became stronger in G₂/M phase. In addition, MARCH5, a mitochondrial E3 ubiquitin ligase, reduced Mfn1 levels and the MARCH5-mediated Mfn1 ubiquitylation were enhanced in G₂/M phase.

Conclusions

Mfn1 is degraded through the MARCH5-mediated ubiquitylation in G₂/M phase and the cell cycle-dependent degradation of Mfn1 could be facilitated by interaction with cyclin B1/Cdk1 complexes.

     

Differential elemental uptake in three pseudo-metallophyte C<Subscript>4</Subscript> grasses in situ in the eastern USA

Background and aims

Elemental uptake in serpentine floras in eastern North America is largely unknown. The objective of this study was to determine major and trace element concentrations in soil and leaves of three native pseudo-metallophyte C₄ grasses in situ at five sites with three very different soil types, including three serpentine sites, in eastern USA.

Methods

Pseudo-total and extractible concentrations of 15 elements were measured and correlated from the soils and leaves of three species at the five sites.

Results

Element concentrations in soils of pseudo-metallophytes varied up to five orders of magnitude. Soils from metalliferous sites exhibited higher concentrations of their characteristic elements than non-metalliferous. In metallicolous populations, elemental concentrations depended on the element. Concentrations of major elements (Ca, Mg, K) in leaves were lower than typical toxicity thresholds, whereas concentrations of Zn were higher.

Conclusions

In grasses, species can maintain relatively low metal concentrations in their leaves even when soil concentrations are richer. However, in highly Zn-contaminated soil, we found evidence of a threshold concentration above which Zn uptake increases drastically. Finally, absence of main characteristics of serpentine soil at one site indicated the importance of soil survey and restoration to maintain serpentinophytes communities and avoid soil encroachment.

     

Engineering a glycerol utilization pathway in Corynebacterium glutamicum for succinate production under O<Subscript>2</Subscript> deprivation

Objective

To explore the glycerol utilization pathway in Corynebacterium glutamicum for succinate production under O₂ deprivation.

Result

Overexpression of a glycerol facilitator, glycerol dehydrogenase and dihydroxyacetone kinase from Escherichia coli K-12 in C. glutamicum led to recombinant strains NC-3G diverting glycerol utilization towards succinate production under O₂ deprivation. Under these conditions, strain NC-3G efficiently consumed glycerol and produced succinate without growth. The recombinant C. glutamicum utilizing glycerol as the sole carbon source showed higher intracellular NADH/NAD⁺ ratio compare with utilizing glucose. The mass conversion of succinate increased from 0.64 to 0.95. Using an anaerobic fed-batch fermentation process, the final strain produced 38.4 g succinate/l with an average yield of 1.02 g/g.

Conclusions

The metabolically-engineered strains showed an efficient succinate production using glycerol as sole carbon source under O₂ deprivation.

     

Low flow extracorporeal CO<Subscript>2</Subscript> removal in ARDS patients: a prospective short-term crossover pilot study

Background

Lung protective mechanical ventilation (MV) is the corner stone of therapy for ARDS. However, its use may be limited by respiratory acidosis.This study explored feasibility of, effectiveness and safety of low flow extracorporeal CO₂ removal (ECCO₂R).

Methods

This was a prospective pilot study, using the Abylcap® (Bellco) ECCO₂R, with crossover off-on-off design (2-h blocks) under stable MV settings, and follow up till end of ECCO₂R. Primary endpoint for effectiveness was a 20% reduction of PaCO₂ after the first 2-h. Adverse events (AE) were recorded prospectively.We included 10 ARDS patients on MV, with PaO₂/FiO₂?<?150 mmHg, tidal volume?≤?8 mL/kg with positive end-expiratory pressure ≥?5 cmH₂O, FiO₂ titrated to SaO₂ 88–95%, plateau pressure ≥?28 cmH₂O, and respiratory acidosis (pH <7.25).

Results

After 2-h of ECCO₂R, 6 patients had a ≥?20% decrease in PaCO₂ (60%); PaCO₂ decreased 28.4% (from 58.4 to 48.7 mmHg, p?=?0.005), and pH increased (1.59%, p?=?0.005). ECCO₂R was hemodynamically well tolerated. During the whole period of ECCO₂R, 6 patients had an AE (60%); bleeding occurred in 5 patients (50%) and circuit thrombosis in 3 patients (30%), these were judged not to be life threatening.

Conclusions

In ARDS patients, low flow ECCO₂R significantly reduced PaCO₂ after 2 h, Follow up during the entire ECCO₂R period revealed a high incidence of bleeding and circuit thrombosis.

Trial registration

https://clinicaltrials.gov identifier: NCT01911533, registered 23 July 2013.

     

Co-cultivation of <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis> with <Emphasis Type="Italic">Azotobacter chroococcum</Emphasis> improved H<Subscript>2</Subscript> production

Objectives

To improve H₂ production, the green algae Chlamydomonas reinhardtii cc849 was co-cultured with Azotobacter chroococcum.

Results

The maximum H₂ production of the co-culture was 350% greater than that of the pure algal cultures under optimal H₂ production conditions. The maximum growth and the respiratory rate of the co-cultures were about 320 and 300% of the controls, and the dissolved O₂ of co-cultures was decreased 74%. Furthermore, the in vitro maximum hydrogenase activity of the co-culture was 250% greater than that of the control, and the in vivo maximum hydrogenase activity of the co-culture was 1.4-fold greater than that of the control. In addition, the maximum starch content of co-culture was 1400% that of the control.

Conclusions

Azotobacter chroococcum improved the H₂ production of the co-cultures by decreasing the O₂ content and increasing the growth and starch content of the algae and the hydrogenase activity of the co-cultures relative to those of pure algal cultures.

     

Comparative effect of inorganic N on plant growth and N<Subscript>2</Subscript> fixation of ten legume crops: towards a better understanding of the differential response among species

Aims

A better understanding of how plant growth, N nutrition and symbiotic nitrogen fixation (SNF) are influenced by soil inorganic N availability, for a wide range of legume species, is crucial to optimise legume productivity, N₂ fixation, while limiting environmental risks such as N leaching.

Methods

A comparative analysis was performed for ten legume crops, grown in a field experiment and supplied with four N fertiliser rates. Dry matter, N concentration and SNF were measured. In parallel, root elongation rates were studied in a greenhouse experiment.

Results

For most species, N fertilisation had little effect on plant growth and N accumulation. SNF was reduced by soil inorganic N available at sowing but with large differences in the magnitude of the response among species. The response varied according to plant N requirements for growth and plant ability to retrieve inorganic N. Accordingly, root lateral expansion rate measured in RhizoTubes was highly correlated with plant ability to retrieve inorganic N measured in the field experiment.

Conclusion

Combining SNF response to soil inorganic N, shoot N and plant ability to retrieve inorganic N, allowed a robust evaluation of differential response to soil inorganic N among a wide range of legume species.