Diversity And Characterization Of Polyhydroxyalkanoate Synthase (Phac) In Seawater And Mangrove Metagenomes

Komuniti mikrob bagi dua tanah paya bakau Pulau Pinang (Batu Maung dan Balik Pulau) yang dipengaruhi oleh aktiviti antropogenik telah dikaji dengan menggunakan pendekatan penjujukan metagenomik “shotgun” tanpa-kultur. Dua set data metagenomik (~250 GB) dihasilkan melalui platfom “Next-generation Sequencing (NGS)” Illumina HiSeq dan disimpan dalam pelayan awam “Metagenomic-Rapid Annotations using Subsystems Technology (MG-RAST)”. Analisis taksonomi mikrob menunjukkan bahawa kedua-dua tanah paya bakau Pulau Pinang didominasi oleh Bakteria (97 %), Proteobakteria (43 %) dan Deltaproteobakteria (15 %) pada peringkat domain,. filum dan kelas masing-masing. Pada peringkat genus, kebanyakan bakteria anaerobik diperhatikan terdiri daripada Deltaproteobakteria. Sebahagian besar daripada jujukan adalah milik spesis mikrob (70 %) dan filum (32 %) yang belum dikenalpasti atau belum dikultur. The microbial communities of two local Penang mangrove soils (Batu Maung and Balik Pulau) which are under anthropogenic influences were investigated using culture-independent shotgun metagenome sequencing approach. Two metagenome data sets (~250 GB) were generated from the Illumina HiSeq next-generation sequencing (NGS) platform and then deposited in Metagenomic-Rapid Annotations using Subsystems Technology (MG-RAST) public server. Microbial taxonomic analysis showed that both Penang mangrove soils were dominated by Bacteria (97 %), Proteobacteria (43 %) and Deltaproteobacteria (15 %) at the domain, phylum and class levels, respectively. At the genus level, predominance of anaerobic bacteria was observed and mostly belonged to Deltaproteobacteria. A large portion of the reads belonged to unknown or yet uncultured microbial species (70 %) and microbial phyla (32 %)

Engineered Mutants of a Marine Photosynthetic Purple Nonsulfur Bacterium with Increased Volumetric Productivity of Polyhydroxyalkanoate Bioplastics

Polyhydroxyalkanoates (PHAs) are green and sustainable bioplastics that could replace petrochemical synthetic plastics without posing environmental threats to living organisms. In addition, sustainable PHA production could be achieved using marine photosynthetic purple nonsulfur bacteria (PNSBs) that utilize natural seawater, sunlight, carbon dioxide gas, and nitrogen gas for growth. However, PHA production using marine photosynthetic PNSBs has not been economically feasible yet due to its high cost and low productivity. In this work, strain improvement, using genome-wide mutagenesis coupled with high-throughput screening via fluorescence-activated cell sorting, we were able to create Rhodovulum sulfidophilum mutants with enhanced volumetric PHA productivity, with an up to 1.7-fold increase. The best selected mutants (E6 and E6M4) reached the stationary growth phase 1 day faster and accumulated the maximum PHA content 2 days faster than the wild type. Maximizing volumetric PHA productivity before the stationary growth phase is indeed an additional advantage for R. sulfidophilum as a growth-associated PHA producer

Identification and phenotypic plasticity of Pseudanabaena catenata from the Svalbard archipelago

A filamentous benthic cyanobacteria, strain USMAC16, was isolated from the High Arctic Svalbard archipelago, Norway, and a combination of morphological, ultrastructural and molecular characterisation (16S rRNA gene sequence) used to identify to species level. Cell dimensions, thylakoid arrangement and apical cell shape are consistent with the Pseudanabaena genus description. The molecular characterisation of P. catenata gave 100% similarity with Pseudanabaena catenata SAG 1464-1, originally reported from Germany. Strain USMAC16 was cultured under a range of temperature and photoperiod conditions, in solid and liquid media, and harvested at exponential phase to examine its phenotypic plasticity. Under different culture conditions, we observed considerable variations in cell dimensions. The longest cell (5.91±0.13 μm) was observed at 15°C under 12:12 light:dark, and the widest cell (3.24±0.06 μm) at 4°C under 12:12 light: dark in liquid media. The study provides baseline data documenting the morphological variation of P. catenata in response to changing temperature regimes

User Preferences and Persona Design for an mHealth Intervention to Support Adherence to Cardiovascular Disease Medication in Singapore: A Multi-Method Study.

BACKGROUND: The use of mobile health (mHealth) has gained popularity globally, including for its use in a variety of health interventions, particularly through short message service (SMS) text messaging. However, there are challenges to the use of mHealth, particularly among older users who have a large heterogeneity in usability and accessibility barriers when using technology. OBJECTIVE: In order to better understand and conceptualize the diversity of users and give insight into their particular needs, we turned to persona creation. Personas are user archetypes created through data generated from multi-method inquiry with actual target users. Personas are an appropriate yet largely underutilized component of current mHealth research. METHODS: Leveraging data from a multi-method study conducted in Singapore with an ethnically diverse population including Chinese, Malay, and Indian participants, we used a proforma to analyze data from the qualitative component (ie, 20 in-depth interviews) and quantitative component (ie, 100 interviewer-guided surveys). We then identified key characteristics, including technology use and preferences as well as adherence factors, to synthesize five personas reflective of persons over the age of 40 years in Singapore with atherosclerotic cardiovascular disease (ASCVD) or ASCVD risk factors, such as hypertension. RESULTS: We present five personas typologized as (1) The Quiet Analog, (2) The Busy Grandparent, (3) The Socializer, (4) The Newly Diagnosed, and (5) The Hard-to-Reach. We report on four key characteristics: health care access, medication adherence, mobile phone technology usage (ie, ownership, access, and utilization), and interest in mHealth. Finally, we provide insights into how these personas may be used in the design and implementation of an mHealth intervention. Our work demonstrates how multi-method data can create biopsychosocial personas that can be used to explore and address the diversity in behaviors, preferences, and needs in user groups. CONCLUSIONS: With wider adoption of mHealth, it is important that we consider user-centered design techniques and design thinking in order to create meaningful, patient-centered interventions for adherence to medications. Future research in this area should include greater exploration of how these five personas can be used to better understand how and when is best to deliver mHealth interventions in Singapore and beyond

A study of soil bacterial communities of fildes peninsula, King George Island (South Shetland Islands), Antarctica

There is sparse information on the bacterial diversity of Filldes Peninsula, King George Island of the maritime Antarctic. A metagenomic approach was used in this study to determine the dominant bacterial population in the soil from the lakes, river and glacier at the Fildes Peninsula, King George Island. Total of ten soil and sediment samples were studied for bacterial diversity using the PCR-DGGE approach targeting for the 165 rRNA gene. About 0.12 to 4.80 μg of DNA was extracted from one gram of soil using the hot enzymatic direct lysis DNA extraction method. Partial 16S rRNA gene product was amplified using the GC357f and 907r primer set, and the amplicon was further resolved using DGGE. Ten to 100- fold dilution of the template or an addition of 10 μg/μI of bovine serum albumin (BSA) in the PCR reaction had successfully reduced the inhibitory effect of the contaminants on the laq polymerase. All the soil samples have different profiles of predominant bands. The predominant bands were excised for sequence determination and bacterial identification. A total of 99 bands and 299 clones were selected from ten locations: Antarctic Lake (AL), GFZ Lake (ZL), Estrellas Lake (EL), Playa Elefantes (PE), Minas River (MR), Collins Glacier (CG), Kitiesh Lake (KL), Selem Lake (BL), Geografos Lake (GL) and oil tank area (OT). After grouping these sequences in different operational taxonomic units (OTUs) (similarity <97%), they were categorized into RDP-designated phylum: Bacteroidetes (27.4%), Proteobacteria (25.7%), Acidobacteria (13.1%), Gemmatimonadetes (4.0%), Firmicutes (4.0%), Actinobacteria (3.4%), Chloroflexi (1.7%), Nitrospira (1.1%), Cyanobacteria (1.1%), WS3 (1.1%), Deionococcus-Thermus (0.6%), Spirochaetes (0.6%) and BRC1 (0.6%). Another 15.4% of the sequences were grouped into unclassified bacteria. Almost 90% of the OTUs have closest relative with uncultured bacterium from the NCBI GenBank database. About 79% of the OTUs had been retrieved in regions which were outside from the Antarctic continent. Acidic soils demonstrated lower diversity of bacteria where two weakly acidic soils PE and OT had the lowest Shannon diversity index. There was no obvious correlation for the changes of bacterial communities between those areas influenced by human activities and less-disturbed by human activities except in the oil tank area. Several phylotypes affiliated with hydrocarbon-degrading bacteria were detected in oil tank area

Optimal iron concentrations for growth-associated polyhydroxyalkanoate biosynthesis in the marine photosynthetic purple bacterium Rhodovulum sulfidophilum under photoheterotrophic condition.

Polyhydroxyalkanoates (PHAs) are a group of natural biopolyesters that resemble petroleum-derived plastics in terms of physical properties but are less harmful biologically to the environment and humans. Most of the current PHA producers are heterotrophs, which require expensive feeding materials and thus contribute to the high price of PHAs. Marine photosynthetic bacteria are promising alternative microbial cell factories for cost-effective, carbon neutral and sustainable production of PHAs. In this study, Rhodovulum sulfidophilum, a marine photosynthetic purple nonsulfur bacterium with a high metabolic versatility, was evaluated for cell growth and PHA production under the influence of various media components found in previous studies. We evaluated iron, using ferric citrate, as another essential factor for cell growth and efficient PHA production and confirmed that PHA production in R. sulfidophilum was growth-associated under microaerobic and photoheterotrophic conditions. In fact, a subtle amount of iron (1 to 2 μM) was sufficient to promote rapid cell growth and biomass accumulation, as well as a high PHA volumetric productivity during the logarithmic phase. However, an excess amount of iron did not enhance the growth rate or PHA productivity. Thus, we successfully confirmed that an optimum concentration of iron, an essential nutrient, promotes cell growth in R. sulfidophilum and also enhances PHA utilization

A marine photosynthetic microbial cell factory as a platform for spider silk production

光合成細菌を用いてクモ糸を作ることに成功 --天然資源を利用した物質生産のモデル微生物--. 京都大学プレスリリース. 2020-07-13.Photosynthetic microorganisms such as cyanobacteria, purple bacteria and microalgae have attracted great interest as promising platforms for economical and sustainable production of bioenergy, biochemicals, and biopolymers. Here, we demonstrate heterotrophic production of spider dragline silk proteins, major ampullate spidroins (MaSp), in a marine photosynthetic purple bacterium, Rhodovulum sulfidophilum, under both photoheterotrophic and photoautotrophic growth conditions. Spider silk is a biodegradable and biocompatible material with remarkable mechanical properties. R. sulfidophilum grow by utilizing abundant and renewable nonfood bioresources such as seawater, sunlight, and gaseous CO2 and N2, thus making this photosynthetic microbial cell factory a promising green and sustainable production platform for proteins and biopolymers, including spider silks

Video consultations in primary and specialist care during the covid-19 pandemic and beyond

Even before the covid-19 pandemic, virtual consultations (also called telemedicine consultations) were on the rise, with many healthcare systems advocating a digital-first approach.1 -7 At the start of the pandemic, many GPs and specialists turned to video consultations to reduce patient flow through healthcare facilities and limit infectious exposures.8 -16 Video and telephone consultations also enable clinicians who are well but have to self-isolate, or who fall into high risk groups and require shielding, to continue providing medical care.17 -19 The scope for video consultations for long term conditions is wide and includes management of diabetes, hypertension, asthma, stroke, psychiatric illnesses, cancers, and chronic pain.20 -22 Video consultations can also be used for triage and management of a wide range of acute conditions, including, for example, emergency eye care triage.23 -25 This practice pointer summarises the evidence on the use of video consultations in healthcare and offers practical recommendations for video consulting in primary care and outpatient settings.Ministry of Health (MOH)Nanyang Technological UniversityJC gratefully acknowledges NTU Singapore's support for Centre for Population Health Sciences which enabled this work. JC's post at Imperial College London is supported by the NIHR NW London Applied Research Collaboration. GC-HK is supported by the Singapore Ministry of Health's National Medical Research Council under the Singapore Population Health Improvement Centre and the Diabetes, Tuberculosis and Neuroscience (Telehealth Core) Centre Grant Programmes

Characterisation of Pseudanabaena amphigranulata (Synechococcales) isolated from a man-made pond, Malaysia: A polyphasic approach

A filamentous benthic cyanobacteria strain isolated from a tropical man-made pond in Malaysia was characterised using combined phenotypic and genetic approaches. Morphological and ultrastructural observations were performed together with growth measurements. Cell dimensions, thylakoid arrangement and apical cell shape with aerotopes were consistent with the description of Pseudanabaena amphigranulata (Goor) Anagnostidis. Molecular characterisation of the16S rRNA gene gave 94% pairwise sequence identity with Pseudanabaena sp. PCC 6802,which corresponds to the genus identification threshold value while also suggesting that the strain is distinctly different to the species of Pseudanabaena currently represented in available databases. The strain showed identical 16S-23S ITS configuration with other strains of Pseudanabaena apart from having a larger spacer region. Cultures of the strain were exposed to various temperature and photoperiod treatments and harvested at exponential phase in order to examine phenotypic plasticity. Significant relationships between environmental conditions and morphological characteristics (cell dimensions and shape) were identified for the first time within the genus Pseudanabaena. The maximum cell length (5.7 ± 0.07 μm) was observed at 25 °C under 12:12 light to dark, while the greatest cell width (3.2 ± 0.11 μm) was also observed at 25 °C but under 16:8 light to dark. The strain showed high plasticity in cell dimensions and shape under different temperature and photoperiod treatments, with25 °C under 12:12 light to dark providing the optimal conditions for its growth

RNA-Seq Analysis Provides Insights for Understanding Photoautotrophic Polyhydroxyalkanoate Production in Recombinant <i>Synechocystis</i> Sp.

<div><p>The photosynthetic cyanobacterium, <i>Synechocystis</i> sp. strain 6803, is a potential platform for the production of various chemicals and biofuels. In this study, direct photosynthetic production of a biopolymer, polyhydroxyalkanoate (PHA), in genetically engineered <i>Synechocystis</i> sp. achieved as high as 14 wt%. This is the highest production reported in <i>Synechocystis</i> sp. under photoautotrophic cultivation conditions without the addition of a carbon source. The addition of acetate increased PHA accumulation to 41 wt%, and this value is comparable to the highest production obtained with cyanobacteria. Transcriptome analysis by RNA-seq coupled with real-time PCR was performed to understand the global changes in transcript levels of cells subjected to conditions suitable for photoautotrophic PHA biosynthesis. There was lower expression of most PHA synthesis-related genes in recombinant <i>Synechocystis</i> sp. with higher PHA accumulation suggesting that the concentration of these enzymes is not the limiting factor to achieving high PHA accumulation. In order to cope with the higher PHA production, cells may utilize enhanced photosynthesis to drive the product formation. Results from this study suggest that the total flux of carbon is the possible driving force for the biosynthesis of PHA and the polymerizing enzyme, PHA synthase, is not the only critical factor affecting PHA-synthesis. Knowledge of the regulation or control points of the biopolymer production pathways will facilitate the further use of cyanobacteria for biotechnological applications.</p></div