Gene editing platforms have revolutionized the field of genetics with a direct impact on the public health system. Although there are apparent benefits, it is often accompanied by public debates over its uncertainties and risks. In the Malaysian context, modern biotechnology has raised questions about how to best govern gene editing in regulations, biosafety, and biosecurity. Even though standards and guidelines on stem cell and cell-based therapies have been developed, there are no appropriate legal frameworks available for gene editing yet. Nevertheless, biosafety regulations were established to balance promoting biotechnology and protecting against their potential environmental and human health risks. There is also a need to address the potential of genetically modified organisms (GMOs) as bioweapons. Numerous frameworks from several international organizations may provide valuable input in formulating documents on gene editing. By establishing comprehensive guidelines, legal policies, and standards to tackle the challenges and risks associated with gene editing, Malaysia can successfully apply this modern technology in this country.
The ever-expanding human population puts tremendous pressure on global food security. With climate change threats lowering crop productivity and food nutritional quality, it is important to search for alternative and sustainable food sources. Microalgae are a promising carbon-neutral biomass with fast growth rate and do not compete with terrestrial crops for land use. More so, microalgae synthesize exclusive marine carotenoids shown to not only exert antioxidant activities but also anti-cancer properties. Unfortunately, the conventional method for fucoxanthin extraction is mainly based on solvent extraction, which is cheap but less environmentally friendly. With the emergence of greener extraction techniques, the extraction of fucoxanthin could adopt these strategies aligned to UN Sustainable Development Goals (SDGs). This is a timely review with a focus on existing fucoxanthin extraction processes, complemented with future outlook on the potential and limitations in alternative fucoxanthin extraction technologies. This review will serve as an important guide to the sustainable and environmentally friendly extraction of fucoxanthin and other carotenoids including but not limited to astaxanthin, lutein or zeaxanthin. This is aligned to the SDGs wherein it is envisaged that this review becomes an antecedent to further research work in extract standardization with the goal of meeting quality control and quality assurance benchmarks for future commercialization purposes.
The search for biodegradable plastics has become the focus in combating the global plastic pollution crisis. Polyhydroxyalkanoates (PHAs) are renewable substitutes to petroleum-based plastics with the ability to completely mineralize in soil, compost, and marine environments. The preferred choice of PHA synthesis is from bacteria or archaea. However, microbial production of PHAs faces a major drawback due to high production costs attributed to the high price of organic substrates as compared to synthetic plastics. As such, microalgal biomass presents a low-cost solution as feedstock for PHA synthesis. Photoautotrophic microalgae are ubiquitous in our ecosystem and thrive from utilizing easily accessible light, carbon dioxide and inorganic nutrients. Biomass production from microalgae offers advantages that include high yields, effective carbon dioxide capture, efficient treatment of effluents and the usage of infertile land. Nevertheless, the success of large-scale PHA synthesis using microalgal biomass faces constraints that encompass the entire flow of the microalgal biomass production, i.e., from molecular aspects of the microalgae to cultivation conditions to harvesting and drying microalgal biomass along with the conversion of the biomass into PHA. This review discusses approaches such as optimization of growth conditions, improvement of the microalgal biomass manufacturing technologies as well as the genetic engineering of both microalgae and PHA-producing bacteria with the purpose of refining PHA production from microalgal biomass.
Green microalgae containing various bioactive compounds and macronutrients such as lipids, carbohydrates, and proteins, have attracted much attention from the global community. Microalgae has the potential to be applied in food industries due to its high protein content, rapid growth rate, and ability to survive in harsh conditions. This study presents a simple yet efficient technique of sonication-assisted triphasic partitioning process, also known as ultrasonic-assisted three phase partitioning (UATPP), for the extraction of proteins from Chlorella vulgaris FSP-E. Comparison studies between three phase partitioning (TPP) and UATPP was conducted to investigate the feasibility of the enhanced technique on proteins extraction. Types of salt, ratio of slurry to t-butanol, salt saturation, sonication frequency, power, irradiation time, and duty cycle as well as biomass loading were studied. UATPP was found to be an improved technique compared to TPP. An optimum separation efficiency and yield of 74.59 ± 0.45 and 56.57 ± 3.70% was obtained, respectively, with the optimized conditions: salt saturation (50%), slurry to t-butanol ratio (1:2), sonication power (100%), irradiation time (10 min), frequency (35 kHz), duty cycle (80%) and biomass loading (0.75 wt%). A scaled-up study was performed to validate the reliability of UATPP for protein extraction. The outcome of the study revealed that UATPP is an attractive approach for downstream processing of microalgae.
Plastics are essential in modern life, but their conventional production is problematic due to environmental pollution and waste management issues. Polylactic acid (PLA) is a widely used bioplastic that is bio-based and biodegradable, making it a key player in the bioeconomy. PLA has been proven to be degradable in various settings, including aqueous, soil, and compost environments. However, monitoring and optimizing PLA biodegradation remains challenging. This study proposes methods to improve the quantification of PLA biodegradation by Amycolatopsis spp. Ultrasound treatments (10 s) significantly improved the enumeration of viable Amycolatopsis cells by breaking the pellets into quantifiable individual cells. A separation technique combining ultrasound (120 s) and 40 μm cell strainers effectively isolated PLA particles from biomass to quantify PLA weight loss. This enabled the monitoring of PLA biofragmentation. Finally, CO2 production was measured according to ISO 14852 to quantify mineralization. Integrating these methods provides an improved quantification for PLA biodegradation along its different stages. In a case study, this led to the construction of a carbon balance where 85.1% of initial carbon content was successfully tracked. The developed techniques for monitoring of PLA biodegradation are essential to design future waste management strategies for biodegradable plastics.
Stem cell-based therapy appears as a promising strategy to induce regeneration of damaged and diseased tissues. However, low survival, poor engraftment and a lack of site-specificity are major drawbacks. Polysaccharide hydrogels can address these issues and offer several advantages as cell delivery vehicles. They have become very popular due to their unique properties such as high-water content, biocompatibility, biodegradability and flexibility. Polysaccharide polymers can be physically or chemically crosslinked to construct biomimetic hydrogels. Their resemblance to living tissues mimics the native three-dimensional extracellular matrix and supports stem cell survival, proliferation and differentiation. Given the intricate nature of communication between hydrogels and stem cells, understanding their interaction is crucial. Cells are incorporated with polysaccharide hydrogels using various microencapsulation techniques, allowing generation of more relevant models and further enhancement of stem cell therapies. This paper provides a comprehensive review of human stem cells and polysaccharide hydrogels most used in regenerative medicine. The recent and advanced stem cell microencapsulation techniques, which include extrusion, emulsion, lithography, microfluidics, superhydrophobic surfaces and bioprinting, are described. This review also discusses current progress in clinical translation of stem-cell encapsulated polysaccharide hydrogels for cell delivery and disease modeling (drug testing and discovery) with focuses on musculoskeletal, nervous, cardiac and cancerous tissues.
Schistosomiasis is one of the neglected tropical diseases that affect millions of people worldwide. Globally, it affects economically poor countries, typically due to a lack of proper sanitation systems, and poor hygiene conditions. Currently, no vaccine is available against schistosomiasis, and the preferred treatment is chemotherapy with the use of praziquantel. It is a common anti-schistosomal drug used against all known species of Schistosoma. To date, current treatment primarily the drug praziquantel has not been effective in treating Schistosoma species in their early stages. The drug of choice offers low bioavailability, water solubility, and fast metabolism. Globally drug resistance has been documented due to overuse of praziquantel, Parasite mutations, poor treatment compliance, co-infection with other strains of parasites, and overall parasitic load. The existing diagnostic methods have very little acceptability and are not readily applied for quick diagnosis. This review aims to summarize the use of nanotechnology in the treatment, diagnosis, and prevention. It also explored safe and effective substitute approaches against parasitosis. At this stage, various nanomaterials are being used in drug delivery systems, diagnostic kits, and vaccine production. Nanotechnology is one of the modern and innovative methods to treat and diagnose several human diseases, particularly those caused by parasite infections. Herein we highlight the current advancement and application of nanotechnological approaches regarding the treatment, diagnosis, and prevention of schistosomiasis.
Ground reaction force (GRF) is a key metric in biomechanical research, including parameters of loading rate (LR), first impact peak, second impact peak, and transient between first and second impact peaks in heel strike runners. The GRFs vary over time during stance. This study was aimed to investigate the variances of GRFs in rearfoot striking runners across incremental speeds. Thirty female and male runners joined the running tests on the instrumented treadmill with speeds of 2.7, 3.0, 3.3, and 3.7 m/s. The discrete parameters of vertical average loading rate in the current study are consistent with the literature findings. The principal component analysis was modeled to investigate the main variances (95%) in the GRFs over stance. The females varied in the magnitude of braking and propulsive forces (PC1, 84.93%), whereas the male runners varied in the timing of propulsion (PC1, 53.38%). The female runners dominantly varied in the transient between the first and second peaks of vertical GRF (PC1, 36.52%) and LR (PC2, 33.76%), whereas the males variated in the LR and second peak of vertical GRF (PC1, 78.69%). Knowledge reported in the current study suggested the difference of the magnitude and patterns of GRF between male and female runners across different speeds. These findings may have implications for the prevention of sex-specific running-related injuries and could be integrated with wearable signals for the in-field prediction and estimation of impact loadings and GRFs.
Malaria is a real and constant danger to nearly half of the world's population of 7.4 billion people. In 2015, 212 million cases were reported along with 429,000 estimated deaths. The World Health Organization recommends artemisinin-based combinatorial therapies, and the artemisinin for this purpose is mainly isolated from the plant Artemisia annua. However, the plant supply of artemisinin is irregular, leading to fluctuation in prices. Here, we report the development of a simple, sustainable, and scalable production platform of artemisinin. The five genes involved in artemisinin biosynthesis were engineered into the moss Physcomitrella patens via direct in vivo assembly of multiple DNA fragments. In vivo biosynthesis of artemisinin was obtained without further modifications. A high initial production of 0.21 mg/g dry weight artemisinin was observed after only 3 days of cultivation. Our study shows that P. patens can be a sustainable and efficient production platform of artemisinin that without further modifications allow for industrial-scale production. A stable supply of artemisinin will lower the price of artemisinin-based treatments, hence become more affordable to the lower income communities most affected by malaria; an important step toward containment of this deadly disease threatening millions every year.
Increasing demands for the supply of biopharmaceuticals have propelled the advancement of metabolic engineering and synthetic biology strategies for biomanufacturing of bioactive natural products. Using metabolically engineered microbes as the bioproduction hosts, a variety of natural products including terpenes, flavonoids, alkaloids, and cannabinoids have been synthesized through the construction and expression of known and newly found biosynthetic genes primarily from model and non-model plants. The employment of omics technology and machine learning (ML) platforms as high throughput analytical tools has been increasingly leveraged in promoting data-guided optimization of targeted biosynthetic pathways and enhancement of the microbial production capacity, thereby representing a critical debottlenecking approach in improving and streamlining natural products biomanufacturing. To this end, this mini review summarizes recent efforts that utilize omics platforms and ML tools in strain optimization and prototyping and discusses the beneficial uses of omics-enabled discovery of plant biosynthetic genes in the production of complex plant-based natural products by bioengineered microbes.
Antioxidants found in microalgae play an essential role in both animals and humans, against various diseases and aging processes by protecting cells from oxidative damage. In this study, 26 indigenous tropical marine microalgae were screened. Out of the 26 screened strains, 10 were selected and were further investigated for their natural antioxidant compounds which include carotenoids, phenolics, and fatty acids collected in their exponential and stationary phases. The antioxidant capacity was also evaluated by a total of four assays, which include ABTS, DPPH, superoxide radical (O2•-) scavenging capacity, and nitric oxide (•NO-) scavenging capacity. This study revealed that the antioxidant capacity of the microalgae varied between divisions, strains, and growth phase and was also related to the content of antioxidant compounds present in the cells. Carotenoids and phenolics were found to be the major contributors to the antioxidant capacity, followed by polyunsaturated fatty acids linoleic acid (LA), eicosapentaenoic acid (EPA), arachidonic acid (ARA), and docosahexaenoic acid (DHA) compared to other fatty acids. The antioxidant capacity of the selected bacillariophytes and haptophytes was found to be positively correlated to phenolic (R2-value = 0.623, 0.714, and 0.786 with ABTS, DPPH, and •NO-) under exponential phase, and to carotenoid fucoxanthin and β-carotene (R2 value = 0.530, 0.581 with ABTS, and 0.710, 0.795 with O2•-) under stationary phase. Meanwhile, antioxidant capacity of chlorophyte strains was positively correlated with lutein, β-carotene and zeaxanthin under the exponential phase (R2 value = 0.615, 0.615, 0.507 with ABTS, and R2 value = 0.794, 0.659, and 0.509 with •NO-). In the stationary phase, chlorophyte strains were positively correlated with violaxanthin (0.755 with •NO-), neoxanthin (0.623 with DPPH, 0.610 with •NO-), and lutein (0.582 with •NO-). This study showed that antioxidant capacity and related antioxidant compound production of tropical microalgae strains are growth phase-dependent. The results can be used to improve the microalgal antioxidant compound production for application in pharmaceutical, nutraceutical, food, and feed industry.
Introduction: A regenerative strategy employing extracellular matrix (ECM)-based biomaterials and stem cells provide a better approach to mimicking the three-dimensional (3D) microenvironment of intervertebral disc for endogenous tissue regeneration. However, there is currently limited understanding regarding the human Wharton Jelly derived-mesenchymal stem cells (hWJ-MSCs) towards nucleus pulposus (NP)-like cells. Our study focused on the development of 3D bioengineered hydrogel based on the predominant ECM of native NP, including type II collagen (COLII) and hyaluronic acid (HA), which aims to tailor the needs of the microenvironment in NP. Methods: We have fabricated a 3D hydrogel using from COLII enriched with HA by varying the biomacromolecule concentration and characterised it for degradation, stability and swelling properties. The WJ-MSC was then encapsulated in the hydrogel system to guide the cell differentiation into NP-like cells. Results: We successfully fabricated COLII hydrogel (2 mg/ml) and HA 10 mg/ml at a weight ratio of HA and COLII at 1:9 and 4.5:9, and both hydrogels physically maintained their 3D sphere-shaped structure after complete gelation. The higher composition of HA in the hydrogel system indicated a higher water intake capacity in the hydrogel with a higher amount of HA. All hydrogels showed over 60% hydrolytic stability over a month. The hydrogel showed an increase in degradation on day 14. The hWJ-MSCs encapsulated in hydrogel showed a round morphology shape that was homogenously distributed within the hydrogel of both groups. The viability study indicated a higher cell growth of hWJ-MSCs encapsulated in all hydrogel groups until day 14. Discussion: Overall, our findings demonstrate that HA/COLII hydrogel provides an optimal swelling capacity, stability, degradability, and non-cytotoxic, thus mimics the NP microenvironment in guiding hWJ-MSCs towards NP phenotype, which is potentially used as an advanced cell delivery system for intervertebral disc regeneration.
Microalgae can use either ammonium or nitrate for its growth and vitality. However, at a certain level of concentration, ammonium nitrogen exhibits toxicity which consequently can inhibit microalgae productivity. Therefore, this study is aimed to investigate the tolerance of Tetraselmis tetrathele to high ammonium nitrogen concentrations and its effects on growth rate, photosynthetic efficiency (F v /F m ), pigment contents (chlorophyll a, lutein, neoxanthin, and β-carotene), and fatty acids production. Experiments were performed at different ammonium nitrogen concentrations (0.31-0.87 gL-1) for 6 days under a light source with an intensity of 300 μmol photons m-2 s-1 and nitrate-nitrogen source as the experimental control. The findings indicated no apparent enhancement of photosynthetic efficiency (Fv/Fm) at high levels of ammonium nitrogen (
NH
4
+
-N) for T. tetrathele within 24 h. However, after 24 h, the photosynthetic efficiency of T. tetrathele increased significantly (p < 0.05) in high concentration of
NH
4
+
-N. Chlorophyll a content in T. tetrathele grown in all of the different
NH
4
+
-N levels increased significantly compared to nitrate-nitrogen (NO3-N) treatment (p < 0.05); which supported that this microalgal could grow even in high level of
NH
4
+
-N concentrations. The findings also indicated that T. tetrathele is highly resistant to high ammonium nitrogen which suggests T. tetrathele to be used in the aquaculture industry for bioremediation purpose to remove ammonium nitrogen, thus reducing the production cost while improving the water quality.
The development of cost-effective, biocompatible soft wound dressings is highly desirable; however, conventional dressings are only designed for flat wounds, which creates difficulty with promising healing efficiency in complex practical conditions. Herein, we developed a tough, adhesive biomimetic hyaluronic acid methacryloyl hydrogels composed of chemically crosslinked hyaluronic acid methacryloyl (HAMA) network and poly(N-hydroxyethyl acrylamide) (PHEAA) network rich in multiple hydrogen bonding. Due to the multiple chemical crosslinking sites (acrylamide groups) of HAMA; the bulk HEMA/PHEAA hydrogels presented significant enhancements in mechanical properties (∼0.45 MPa) than common hyaluronic acid hydrogels (<0.1 MPa). The abundant hydrogen bonding also endowed the resultant hydrogels with extremely high adhesiveness on many nonporous substrates, including glass and biological tissues (e.g., heart, liver, lung, kidney, stomach, and muscle), with a considerable interfacial toughness of ∼1432 J m-2. Accordingly, since both natural hyaluronic acid derivative polymers and hydrophilic PHEAA networks are highly biocompatible, the hydrogel matrix possesses good blood compatibility (<5% of hemolysis ratio) and satisfies the general dressing requirements (>99% of cell viability). Based on these physicochemical features, we have demonstrated that this adhesive hydrogel, administered in the form of a designed patch, could be applied to wound tissue healing by promoting epithelialization, angiogenesis, and collagen deposition. We believe that our proposed biomimetic hydrogel design holds great potential for wound repair and our developed HAMA/PHEAA hydrogels are extremely promising for the next-generation tissue healings in emergency situations.