Biosynthesis of apigenin glucosides in engineered Corynebacterium glutamicum.

Glucosylation is a well-known approach to improve the solubility, pharmacological, and biological properties of flavonoids, making flavonoid glucosides a target for large-scale biosynthesis. However, the low yield of products coupled with the requirement of expensive UDP-sugars limits the application of enzymatic systems for large-scale. C. glutamicum is a Gram-positive and generally regarded as safe (GRAS) bacteria frequently employed for the large-scale production of amino acids and bio-fuels. Due to the versatility of its cell factory system and its non-endotoxin producing properties, it has become an attractive system for the industrial-scale biosynthesis of alternate products. Here, we explored the cell factory of C. glutamicum for efficient glucosylation of flavonoids using apigenin as a model flavonoid, with the heterologous expression of a promiscuous glycosyltransferase, YdhE from Bacillus licheniformis and the endogenous overexpression of C. glutamicum genes galU1 encoding UDP-glucose pyrophosphorylase and pgm encoding phosphoglucomutase involved in the synthesis of UDP-glucose to create a C. glutamicum cell factory system capable of efficiently glucosylation apigenin with a high yield of glucosides production. Consequently, the production of various apigenin glucosides was controlled under different temperatures yielding almost 4.2 mM of APG1(apigenin-4'-O-ß-glucoside) at 25℃, and 0.6 mM of APG2 (apigenin-7-O-ß-glucoside), 1.7 mM of APG3 (apigenin-4',7-O-ß-diglucoside) and 2.1 mM of APG4 (apigenin- 4',5-O-ß-diglucoside) after 40 h of incubation with the supplementation of 5 mM of apigenin and 37℃. The cost-effective developed system could be used to modify a wide range of plant secondary metabolites with increased pharmacokinetic activities on a large scale without the use of expensive UDP-sugars.

Berberine-loaded polylactic acid nanofiber scaffold as a drug delivery system: The relationship between chemical characteristics, drug-release behavior, and antibacterial efficiency.

Hydrophobic berberine powder (BBR) and hydrophilic BBR nanoparticles (BBR NPs) were loaded into an electrospun polylactic acid (PLA) nanofiber scaffold for modulating the release behavior of BBR in an aqueous medium. The BBR release from the BBR/PLA and BBR NPs/PLA nanofiber scaffolds was investigated in relation to their chemical characteristics, BBR dispersion into nanofibers, and wettability. The BBR release profiles strongly influenced the antibacterial efficiency of the scaffolds over time. When the BBR was loaded, the BBR/PLA nanofiber scaffold exhibited an extremely hydrophobic feature, causing a triphasic release profile in which only 9.8 wt % of the loaded BBR was released in the first 24 h. This resulted in a negligible inhibitory effect against methicillin-resistant Staphylococcus aureus bacteria. Meanwhile, the BBR NPs/PLA nanofiber scaffold had more wettability and higher concentration of BBR NPs dispersed on the surface of PLA nanofibers. This led to a sustained release of 75 wt % of the loaded BBR during the first 24 h, and consequently boosted the antibacterial effectiveness. Moreover, the cytotoxicity test revealed that the BBR NPs/PLA nanofiber scaffold did not induce any changes in morphology and proliferation of MA-104 cell monolayers. It suggests that the BBR/PLA and BBR NPs/PLA nanofiber scaffolds can be used in different biomedical applications, such as wound dressing, drug delivery systems, and tissue engineering, according to the requirement of BBR concentration for the desired therapeutic effects.

The role of chitosan in enhancing the solubility and antibacterial activity of emodin against drug-resistant bacteria.

Similar to most anthraquinone compounds, the pharmacological properties of emodin are limited because of its low water solubility. In this study, the formulation of chitosan and emodin (EMD/CS) was prepared by a bottom-up method with precipitation and sonication steps in order to enhance the solubility of emodin. Thanks to the interactions of oxygen-and nitrogen-containing groups in chitosan with emodin molecules, the solubility of emodin in the formulation was remarkably increased to 0.5 mg/mL. The EMD/CS particles were well dispersed and distributed in a range of sub-micrometer with an average particle size of 342 nm. The EMD/CS formulation exhibited synergic antibacterial activity of emodin and chitosan, against drug-resistant bacterial strains, namely Methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli O157:H7 (E. coli O157:H7). When the compositions of emodin and chitosan increased, the antibacterial effectiveness of the EMD/CS formulation increased. The EMD/CS formulation with compositions of 0.5 mg/mL of emodin and 9.0 mg/mL of chitosan could significantly inhibit the proliferation of E. coli O157:H7. Meanwhile, the EMD/CS formulation with a lower concentration of emodin (0.4 mg/mL) and chitosan (7.2 mg/mL) could cause an extermination effect on MRSA. The enhanced solubility of EMD/CS formulation suggests that this formulation can be a potential candidate for the treatment of infectious diseases caused by drug-resistant bacterial pathogens.

Broadband optical vortex beam generation using flat-surface nanostructured gradient index vortex phase masks.

We developed a new kind of compact flat-surface nanostructured gradient index vortex phase mask, for the effective generation of optical vortex beams in broadband infrared wavelength range. A low-cost nanotechnological material method was employed for this work. The binary structure component consists of 17,557 nano-sized rods made of two lead-bismuth-gallium silicate glasses which were developed in-house. Those small rods are spatially arranged in such a way that, according to effective medium theory, the refractive index of this internal structure is constant in the radial direction and linearly changes following azimuthal angle. Numerical results demonstrated that a nanostructured vortex phase mask with a thickness of 19 µm can convert Gaussian beams into fundamental optical vortices over 290 nm wavelength bandwidth from 1275 to 1565 nm. This has been confirmed in experiments using three diode laser sources operating at 1310, 1550, and 1565 nm. The generation of vortex beams is verified through their uniform doughnut-like intensity distributions, clear astigmatic transformation patterns, and spiral as well as fork-like interferograms. This new flat-surface component can be directly mounted to an optical fiber tip for simplifying vortex generator systems as well as easier manipulation of the generated OVB in three-dimensional space.

The diagnostic performance of AFP and PIVKA-II models for non-B non-C hepatocellular carcinoma.

OBJECTIVE: This study aims to describe the diagnostic performance of alpha-fetoprotein (AFP), alpha-fetoprotein L3 isoform (AFP-L3), protein induced by vitamin K absence II (PIVKA-II), and combined biomarkers for non-B non-C hepatocellular carcinoma (NBNC-HCC). RESULTS: A total of 681 newly-diagnosed primary liver disease subjects (385 non-HCC, 296 HCC) who tested negativity for the hepatitis B surface antigen (HBsAg) and hepatitis C antibody (anti-HCV) enrolled in this study. At the cut-off point of 3.8 ng/mL, AFP helps to discriminate HCC from non-HCC with an area under the curve (AUC) value of 0.817 (95% confidence interval [CI]: 0.785-0.849). These values of AFP-L3 (cut-off 0.9%) and PIVKA-II (cut-off 57.7 mAU/mL) were 0.758 (95%CI: 0.725-0.791) and 0.866 (95%CI: 0.836-0.896), respectively. The Bayesian Model Averaging (BMA) statistic identified the optimal model, including patients' age, aspartate aminotransferase, AFP, and PIVKA-II combination, which helps to classify HCC with better performance (AUC = 0.896, 95%CI: 0.872-0.920, P < 0.001). The sensitivity and specificity of the optimal model reached 81.1% (95%CI: 76.1-85.4) and 83.2% (95%CI: 78.9-86.9), respectively. Further analyses indicated that AFP and PIVKA-II markers and combined models have good-to-excellent performance detecting curative resected HCC, separating HCC from chronic hepatitis, dysplastic, and hyperplasia nodules.

Glucosylation of Isoeugenol and Monoterpenes in Corynebacterium glutamicum by YdhE from Bacillus lichenformis.

Corynebacterium glutamicum has been regarded as a food-grade microorganism. In recent years, the research to improve the activities of beneficial therapeutics and pharmaceutical substances has resulted in the engineering of the therapeutically favorable cell factory system of C. glutamicum. In this study, we successfully glucosylated isoeugenol and other monoterpene derivatives in C. glutamicum using a promiscuous YdhE, which is a glycosyltransferase from Bacillus lichenformis. For efficient glucosylation, cultivation conditions such as the production time, substrate concentration, carbon source, and culture medium were optimized. Our system successfully converted about 93% of the isoeugenol to glucosylated compounds in the culture. The glucoside compounds were then purified, analyzed, and identified as isoeugenol-1-O-ß-d-glucoside and isoeugenol-1-O-ß-d-(2″-acetyl)-glucoside.

Complexes of Ag and ZnO nanoparticles with BBR for enhancement of gastrointestinal antibacterial activity through the impacts of size and composition.

This study introduces the bioformulations of Ag/BBR and ZnO/BBR complexes against pathogenic bacteria in the gastrointestinal tract. Without the use of toxic reduction agents, Ag and ZnO NPs were prepared using an electrochemical method and then facially mixed with BBR solution to form Ag/BBR and ZnO/BBR complexes. BBR molecules are strongly conjugated with Ag and ZnO NPs through coordinated bonding and electrostatic interaction. As a result, the presence of BBR significantly influenced the nanoparticle growth, resulting in the formation of core/shell structured Ag/BBR and ZnO/BBR NPs with small particle sizes. The antibacterial test showed that BBR, Ag, or ZnO components all contributed to the increase of antibacterial ability of Ag/BBR and ZnO/BBR NPs against both methicillin-resistant Staphylococcus aureus (MRSA) and Salmonella enteritidis (S. enteritidis). The bactericidal ability of Ag/BBR and ZnO/BBR complexes against MRSA was exhibited even at a concentration of four-fold dilution (corresponding to 1.25 g L-1 of BBR and 46.25 mg L-1 of Ag) and two-fold dilution (corresponding to 2.5 g L-1 of BBR and 10 mg L-1 of ZnO), respectively, while that of the Ag/BBR complex against S. enteritidis showed at a concentration of two-fold dilution corresponding to 2.5 g L-1 of BBR and 92.5 mg L-1 of Ag. The results obtained in this study support that Ag/BBR and ZnO/BBR complexes can be potential therapeutic agents against gastrointestinal infections.

Plastic waste in sandy beaches and surface water in Thanh Hoa, Vietnam: abundance, characterization, and sources.

The occurrence and characterization of marine debris on beaches bring opportunities to track back the anthropogenic activities around shorelines as well as aid in waste management and control. In this study, the three largest beaches in Thanh Hoa (Vietnam) were examined for plastic waste, including macroplastics (≥ 5 mm) on sandy beaches and microplastics (MPs) (< 5 mm) in surface water. Among 3803 items collected on the beaches, plastic waste accounted for more than 98%. The majority of the plastic wastes found on these beaches were derived from fishing boats and food preservation foam packaging. The FT-IR data indicated that the macroplastics comprised 77% polystyrene, 17% polypropylene, and 6% high-density polyethylene, while MPs discovered in surface water included other forms of plastics such as polyethylene- acrylate, styrene/butadiene rubber gasket, ethylene/propylene copolymer, and zein purified. FT-IR data demonstrated that MPs might also be originated from automobile tire wear, the air, and skincare products, besides being degraded from macroplastics. The highest abundance of MPs was 44.1 items/m3 at Hai Tien beach, while the lowest was 15.5 items/m3 at Sam Son beach. The results showed that fragment form was the most frequent MP shape, accounting for 61.4 ± 14.3% of total MPs. MPs with a diameter smaller than 500 µm accounted for 70.2 ± 7.6% of all MPs. According to our research, MPs were transformed, transported, and accumulated due to anthropogenic activities and environmental processes. This study provided a comprehensive knowledge of plastic waste, essential in devising long-term development strategies in these locations.

Biosynthetic pathway of peucemycin and identification of its derivative from Streptomyces peucetius.

Streptomyces peucetius ATCC 27952 is a well-known producer of important anticancer compounds, daunorubicin and doxorubicin. In this study, we successfully identified a new macrolide, 25-hydroxy peucemycin, that exhibited an antibacterial effect on some pathogens. Based on the structure of a newly identified compound and through the inactivation of a polyketide synthase gene, we successfully identified its biosynthetic gene cluster which was considered to be the cryptic biosynthetic gene cluster. The biosynthetic gene cluster spans 51 kb with 16 open reading frames. Five type I polyketide synthase (PKS) genes encode eight modules that synthesize the polyketide chain of peucemycin before undergoing post-PKS tailoring steps. In addition to the regular starter and extender units, some modules have specificity towards ethylmalonyl-CoA and unusual butylmalonyl-CoA. A credible explanation for the specificity of the unusual extender unit has been searched for. Moreover, the enzyme responsible for the final tailoring pathway was also identified. Based on all findings, a plausible biosynthetic pathway is here proposed. KEY POINTS: • Identification of a new macrolide, 25-hydroxy peucemycin. • An FMN-dependent monooxygenase is responsible for the hydroxylation of peucemycin. • The module encoded by peuC is unique to accept the butylmalonyl-CoA as an unusual extender unit.

Emission and distribution profiles of polycyclic aromatic hydrocarbons in solid residues of municipal and industrial waste incinerators, Northern Vietnam.

The concentrations and profiles of 18 polycyclic aromatic hydrocarbons (PAHs) in particulate matter (PM10), fly ash (FA), and bottom ash (BA) were examined in three incineration residues. Samples were collected from different municipal and industrial solid waste incinerators in Northern Vietnam. The average concentrations of total PAHs in PM10, fly ash, and bottom ash were 9.55 × 103 ng/Nm3, 215 × 103 ng/g, and 2.38 ng/g, respectively. Low-molecular-weight PAHs (2 to 3 rings) were predominant in most samples. The emission factor of total PAHs decreased in the order of FA > BA > PM10. A higher concentration of total PAHs was found in industrial facilities than that in municipal ones. The high carcinogenic proportion of PAHs together with significantly high annual emissions reflect the high pollution risk to the ecosystem by PAHs in the case of reuse of incineration ashes (e.g., brick production). Regarding the carcinogenic risk of PAH-bounded ashes or particles, calculations from this study imply the significant threat for workers who have been manipulated in the incineration facilities, directly exposed to fly and bottom ashes. Meanwhile, the risk from PAH-bound particulate was not considered a significant threat for both normal adults and children. Further study on PAHs contained in incinerator waste dumps should be conducted in Vietnam to assess the potential contamination risk of these incineration by-products.

Transmission of an optical vortex beam in antiresonant fibers generated in an all-fiber system.

We report an experimental study on transmission of orbital angular momentum mode in antiresonant fibers generated with a dedicated all-fiber optical vortex phase mask. The vortex generator can convert Gaussian beam into vortex beams with topological charge l = 1. Generated vortex beam is directly butt-coupled into the antiresonant fiber and propagates over distance of 150 cm. The stability and sensitivity of the transmitted vortex beam on the external perturbations including bending, axial stress, and twisting is investigated. We demonstrate distortion-free vortex propagation for the axial stress force below 0.677 N, a bend radius greater than 10 cm.

Antibacterial activity of a berberine nanoformulation.

This study describes the preparation of berberine (BBR) in nanoformulation to enhance its solubility and increase its antibacterial effectiveness against hospital-acquired infections. BBR nanoparticles (BBR NPs) were formed by antisolvent precipitation (ASP) using glycerol as a safe organic solvent. UV-vis absorption spectra demonstrated that the solubility of BBR NPs was greatly enhanced compared to that of pure BBR. Glycerol played a role as a stabilizer for BBR NPs through the formation of hydrogen bonds between glycerol and BBR NPs. The prepared BBR NPs have a narrow size distribution with an average diameter of 156 nm at a concentration of 2.0 mg/mL, measured by dynamic light scattering. After nanoformulation, the concentration of BBR NPs could reach up to 5.0 mg/mL, which is much higher than the saturation concentration without treatment. Results show a strongly enhanced antibacterial activity of BBR NPs compared with that of pure BBR at the same concentration. The minimum bactericidal concentration of BBR NPs against methicillin-resistant Staphylococcus aureus and Escherichia coli O157:H7 was found to be 2.0 and 5.0 mg/mL, respectively. Transmission electron microscopy showed that BBR NPs surrounded the bacterial cells and severely damaged the cell walls. Therefore, BBR NPs prepared by ASP appear to be a potential candidate for the treatment of bacterial pathogens.

Ophthalmologists' and patients' perspectives on treatments for diabetic retinopathy and maculopathy in Vietnam: a descriptive qualitative study.

INTRODUCTION: Globally, diabetic retinopathy (DR) is the leading cause of blindness in working-aged adults. Early detection and treatment of DR is essential for preventing sight loss. Services must be available, accessible and acceptable to patients if we are to ensure they seek such care. OBJECTIVES: To understand patients' knowledge and attitudes towards laser versus antivascular endothelial growth factor (VEGF) injections to treat DR in Vietnam, and to identify factors Vietnamese ophthalmologists consider when making treatment decisions. METHODS: This is a descriptive qualitative study based on semi-structured interviews with 18 patients (12 from Ho Chi Minh City and 6 from Hanoi) plus individual interviews with 24 ophthalmologists working in eye clinics in these cities. Thematic analysis was used to analyse the data. RESULTS: In total, 10/24 (41.7%) ophthalmologists were female, and their median age was 41 years (range 29-69 years). The median age of patients was 56.5 years (range 28-72 years), and 7/18 (38.9%) were female. Briefly, factors that influence DR treatment decisions for ophthalmologists are medical considerations (ie, severity of disease, benefits and risks), availability (ie, treatment and resources) and patient-related factors (ie, costs and adherence). Patient's perceived barriers and facilitators to treatments were based on patient and family related factors (ie, treatment and transportation costs) and previous treatment experiences (ie, positive and negative). Recommendations by all participants included ensuring that both laser and anti-VEGF injections are widely available across the country and controlling costs for patients and the healthcare system. CONCLUSIONS: Reducing DR treatment costs, optimising treatments options, and expanding the network of clinics offering treatment outside metropolitan areas were the main issues raised by participants. These findings can help inform policy changes in Vietnam and may be generalisable to other low-resource settings.

Predicting Anxiety and Depression Among Patients With COVID-19 in Concentrated Isolation at Medical Camps in Vietnam: A Descriptive Cross-Sectional Study.

Introduction: This study aims to assess the requirement for anxiety and depression treatment for patients with coronavirus disease 2019 (COVID-19) in medical camps in Bac Giang province, Vietnam. This information can help improve the government policy to reduce anxiety and depression in patients with COVID-19. Methods: A total of patients with 427 COVID-19 participated in the survey conducted from 5 to 15 June 2021 in Bac Giang province. The survey included 17 questions about the general characteristics of the patients, 15 questions to assess common COVID-19 symptoms, the Patient Health Questionnaire-9 (PHQ-9), and General Anxiety Disorder-7 (GAD-7) scores, and four questions to assess hospital reviews, including facilities, food, medical staff, and living conditions. Logistics regression analyses were conducted to assess the association between COVID-19 symptoms and high anxiety and depression (HAD) status. Results: A logistic regression analysis evaluated the risk factors in need of intervention. Our study showed that lower hospital review scores (odd ratio = 0.98; 95% confident interval = 0.97-0.99) were found to be a risk needing intervention. It was also identified that older patients (odd ratio = 1.1; 95% confident interval = 1.03-1.18), women (odd ratio = 1.31; 95% confident interval = 1.09-1.31), patients who were primary income earners in the family (odd ratio = 1.15; 95% confident interval = 1.03-1.28), patients who had headaches (odd ratio = 1.16; 95% confident interval = 1.06-1.21), and patients who had joint pain (odd ratio = 1.17; 95% confident interval = 1.06- 1.3) were risk factors for HAD status. Conclusion: Our research shows that every 10-year age increase was associated with a 10% increase in the likelihood of HAD status. Study subjects being primary income earners were also associated with a 15% increased risk of having HAD status. This study showed that a decrease in family income due to COVID-19 caused an increase in high-level anxiety/depression status.

N-Glucosylation in Corynebacterium glutamicum with YdhE from Bacillus lichenformis.

Corynebacterium glutamicum is traditionally known as a food-grade microorganism due to its high ability to produce amino acids and its endotoxin-free recombinant protein expression factory. In recent years, studies to improve the activities of useful therapeutics and pharmaceutical compounds have led to the engineering of the therapeutically advantageous C. glutamicum cell factory system. One of the well-studied ways to improve the activities of useful compounds is glucosylation with glycosyltransferases. In this study, we successfully and efficiently glycosylated therapeutic butyl-4-aminobenzoate and other N-linked compounds in C. glutamicum using a promiscuous YdhE, which is a glycosyltransferase from Bacillus lichenformis. For efficient glucosylation, components, such as promoter, codons sequence, expression temperatures, and substrate and glucose concentrations were optimized. With glucose as the sole carbon source, we achieved a conversion rate of almost 96% of the glycosylated products in the culture medium. The glycosylated product of high concentration was successfully purified by a simple purification method, and subjected to further analysis. This is a report of the in vivo cultivation and glucosylation of N-linked compounds in C. glutamicum.

Insights Into the Genetics of the Zhonghua 11 Resistance to Meloidogyne graminicola and Its Molecular Determinism in Rice.

Meloidogyne graminicola is a widely spread nematode pest of rice that reduces crop yield up to 20% on average in Asia, with devastating consequences for local and global rice production. Due to the ban on many chemical nematicides and the recent changes in water management practices in rice agriculture, an even greater impact of M. graminicola can be expected in the future, stressing the demand for the development of new sustainable nematode management solutions. Recently, a source of resistance to M. graminicola was identified in the Oryza sativa japonica rice variety Zhonghua 11 (Zh11). In the present study, we examine the genetics of the Zh11 resistance to M. graminicola and provide new insights into its cellular and molecular mechanisms. The segregation of the resistance in F2 hybrid populations indicated that two dominant genes may be contributing to the resistance. The incompatible interaction of M. graminicola in Zh11 was distinguished by a lack of swelling of the root tips normally observed in compatible interactions. At the cellular level, the incompatible interaction was characterised by a rapid accumulation of reactive oxygen species in the vicinity of the nematodes, accompanied by extensive necrosis of neighbouring cells. The expression profiles of several genes involved in plant immunity were analysed at the early stages of infection during compatible (susceptible plant) and incompatible (resistant plant) interactions. Notably, the expression of OsAtg4 and OsAtg7, significantly increased in roots of resistant plants in parallel with the cell death response, suggesting that autophagy is activated and may contribute to the resistance-mediated hypersensitive response. Similarly, transcriptional regulation of genes involved in hormonal pathways in Zh11 indicated that salicylate signalling may be important in the resistance response towards M. graminicola. Finally, the nature of the resistance to M. graminicola and the potential exploitation of the Zh11 resistance for breeding are discussed.

Development of gradient index microlenses for the broadband infrared range.

The development of gradient index free-form micro-optic components dedicated to the mid-infrared range is challenging due to the lack of appropriate technology. We propose a method for developing gradient index components for broadband infrared range beyond the transmission window of silicate glass based on nanostructurization using a stack-and-draw fiber drawing technique. A proof-of-concept microlens is developed and verified experimentally in the wavelength range 1.5-4.3 µm. The microlenses are composed of a set of nanorods with a diameter of 940 nm made of a pair of SiO2-PbO-Bi2O3-Ga2O3 based glasses ordered into the preliminary calculated binary pattern. The pattern forms effectively continuous parabolic refractive index distribution for infrared range according to Maxwell-Garnett effective medium model. The development of individual microlenses with a diameter of 118 µm and focal length of 278 µm at the wavelength of 3.75 µm are reported. A large array of 737 microlenses with an individual diameter of 125 µm and focal length of 375 µm is also presented and analyzed.

Impact of targeted diabetic retinopathy training for graders in Vietnam and the implications for future diabetic retinopathy screening programmes: a diagnostic test accuracy study.

OBJECTIVES: To compare the accuracy of trained level 1 diabetic retinopathy (DR) graders (nurses, endocrinologists and one general practitioner), level 2 graders (midlevel ophthalmologists) and level 3 graders (senior ophthalmologists) in Vietnam against a reference standard from the UK and assess the impact of supplementary targeted grader training. DESIGN: Diagnostic test accuracy study. SETTING: Secondary care hospitals in Southern Vietnam. PARTICIPANTS: DR training was delivered to Vietnamese graders in February 2018 by National Health Service (NHS) UK graders. Two-field retinal images (412 patient images) were graded by 14 trained graders in Vietnam between August and October 2018 and then regraded retrospectively by an NHS-certified reference standard UK optometrist (phase I). Further DR training based on phase I results was delivered to graders in November 2019. After training, a randomised subset of images from January to October 2020 (115 patient images) was graded by six of the original cohort (phase II). The reference grader regraded all images from phase I and II retrospectively in masked fashion. PRIMARY AND SECONDARY OUTCOME MEASURES: Sensitivity was calculated at the two different time points, and χ2 was used to test significance. RESULTS: In phase I, the sensitivity for detecting any DR for all grader groups in Vietnam was low (41.8-42.2%) and improved in phase II after additional training was delivered (51.3-87.2%). The greatest improvement was seen among level 1 graders (p<0.001), and the lowest improvement was observed among level 3 graders (p=0.326). There was a statistically significant improvement in sensitivity for detecting referable DR and referable diabetic macular oedema between all grader levels. The post-training values ranged from 40.0 to 61.5% (including ungradable images) and 55.6%-90.0% (excluding ungradable images). CONCLUSIONS: This study demonstrates that targeted training interventions can improve accuracy of DR grading. These findings have important implications for improving service delivery in DR screening programmes in low-resource settings.

A Simple Method to Produce Engineered Cartilage from Human Adipose-Derived Mesenchymal Stem Cells and Poly ε-Caprolactone Scaffolds.

INTRODUCTION: The damaged articular cartilage has limited self-regeneration capacity because of the absence of blood vessels, lymphatics, and nerves. Cartilage transplantation is, hence, a popular method used to treat this disease. However, sources of autograft and allogenic cartilage for transplantation are limited. Therefore, this study aims to suggest a simple method to produce engineered cartilage from human adipose-derived mesenchymal stem cells (ADSCs) and poly (ε-caprolactone) (PCL) scaffolds. METHODS: ADSCs were isolated and expanded from fat tissues according to published protocols. PCL-porous scaffolds were produced from PCL with 5 × 5 × 0.6 mm3 with 200-400 µ m pore sizes. ADSCs were seeded on the PCL scaffolds at three different densities (104, 105, 106 cells per scaffold). The adherence of ADSCs on the surface of PCL scaffolds was evaluated based on an immunostaining assay to determine the presence of ADSCs. The cell proliferation on PCL scaffolds was determined by MTT assay. The complexity in ADSCs and PCL scaffolds was induced to cartilage using a chondrogenesis medium. The engineered cartilage was characterized by the accumulation of proteoglycan and aggrecan by Safranin O staining assay. Their structures were evaluated using an H-E staining assay. Finally, these engineered cartilage tissues were transplanted into mice to assess cartilage maturation when compared to natural cartilage. RESULTS: The results showed that the engineered cartilage tissues could be successfully produced by cultures of ADSCs on poly ε-caprolactone scaffolds in combination with chondrogenesis medium. The suitable density of ADSCs was 106 cells/per scaffold of 5 × 6 × 0.6 mm3 with pore size from 200 to 400 µ m. CONCLUSION: The results showed that an in vitro cartilage tissue was created from ADSCs and PCL scaffold. The cartilage tissue exists in the mice for 6 months.

Silica-based photonic crystal fiber infiltrated with 1,2-dibromoethane for supercontinuum generation.

This study proposes a photonic crystal fiber (PCF) made of fused silica glass with the core infiltrated with 1,2-dibromoethane (C2H4Br2) as a new source of supercontinuum light pulses. Due to the modifications of the PCF's structure geometry, a number of computer simulations investigating their optimized structures has been carried out. This aimed at achieving flat near-zero dispersion and zero dispersion wavelength matching of the pump wavelength for efficient spectral broadening. Based on the obtained results, the structural geometries of two C2H4Br2-core PCFs were optimized using numerical modeling for broadband supercontinuum (SC) generation. The first fiber structure with a lattice constant 1.5 µm and filling factor 0.4 has all-normal dispersion profile. The SC with a broadened spectral bandwidth from 0.64 to 1.70 µm is generated by pump pulses centered at a wavelength of 1.03 µm, 120 fs duration, and energy of 1.5 nJ. The second proposed structure-with lattice constant 1.5 µm and filling factor 0.65-has anomalous dispersion for wavelengths longer than 1.03 µm. We obtained high coherence of the SC pulses in the anomalous dispersion range over wavelengths of 0.7-2.4 µm with the same pump pulse as the first fiber and with input energy of 0.09 nJ. These fibers would be interesting candidates for all-fiber SC sources operating with low-energy pump lasers as cost-effective alternatives to glass core fibers.