The Hibiscus sabdariffa var. UMKL (Roselle) investigated here may potentially be used as an alternative fibre source. To
the best of our knowledge, there was no study focusing on the genetics underlying the cellulose biosynthesis machinery
in Roselle thus far. This paper presents the results of the first isolation of the cellulose synthase gene, HsCesA1 from this
plant, which is fundamental for working towards understanding the functions of CesA genes in the cellulose biosynthesis
of Roselle. A full-length HsCesA1 cDNA of 3528 bp in length (accession no: KJ608192) encoding a polypeptide of 974
amino acid was isolated. The full-length HsCesA1 gene of 5489 bp length (accession no: KJ661223) with 11-introns
and a promoter region of 737 bp was further isolated. Important and conserved characteristics of a CesA protein were
identified in the HsCesA1 deduced amino acid sequence, which strengthened the prediction that the isolated gene being
a cellulose synthase belonging to the processive class of the 2-glycosyltransferase family 2A. Relative gene expression
analysis by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) on young leaf and stem tissues
found that HsCesA1 had similar levels of gene expression in both tissues. Phylogenetic and Blast analyses also supported
the prediction that the isolated HsCesA1 may play roles in the cell wall depositions in both leaf and stem tissues.
Lignocellulose is the most abundant renewable resource naturally available with great potential for bioconversion to value-added bio-products in this case from coconut husk to reducing sugar. This study was to isolate the lignocellulose bioconversion bacteria from cow dung and termite gut for reducing sugar production. CMC and Congo red dye was used as a selective media to screen for cellulose degrading bacteria while alkaline lignin media used to screen for lignin degrading bacteria. Coconut husk was pretreated using alkaline precipitation method. The 16s rRNA sequence analysis, Gram staining, motility test, crystal formation test and penicillin sensitivity test were used to confirm the bacteria identity. The reducing sugars production was monitor using 3, 5-dinitrosalicyclic acid (DNS) method. Bacterial enzymatic hydrolysis were facilitated through fermentation of 5 locally isolated bacteria at 6 shaking regimes which were 0rpm, 50rpm, 100rpm, 150rpm, 200rpm and 250rpm and conditions were fixed at pH7.0 and temperature 25oC. The total of 39 and 28 colonies were isolated from cow dung and termite gut respectively. The isolates T10, T19, T22, T24, C19, and C37 were identified as Bacillus thuringiensis, Enterobacter aerogenes, Bacillus pumilus, Bacillus vireti, Bacillus anthracis, and Bacillus thuringiensis. In PC medium, 2 bacteria strains, T19 and C37 showed the highest concentration of reducing sugar production (after 18 to 20 hours) which was 1.28 g/L at 150rpm (T19) and 1.27 g/L at 200rpm (C37). While in CMC medium, T19 and C37 also showed the highest concentration of reducing sugar production which was 0.72 g/L at 150rpm (T19) and 0.70 g/L at 200rpm (C37). These bacteria demonstrated significant (p < 0.05) to degrade cellulose was shown by measuring the diameter of clearing zone signifying metabolism of cellulose and also the ability to convert both PC and CMC to reducing sugars measurable by DNS method. This study suggests that the PC resulted in higher reducing sugar production compare to CMC at 150rpm and 200rpm by E. aerogenes and B. thuringiensis which these bacteria isolated from cow dung and termite gut. Future studies on metabolic pathway and specific reducing sugar produced may be done using High Performance Liquid Chromatography (HPLC) analysis.
Each year, influenza A infections have caused tremendous death rate as high as 300,000-500,000 globally. Although
there are effective anti-influenza agents and vaccines, high mutational rate among influenza A viruses renders dramatic
decline in the effectiveness of anti-influenza agents or vaccines in certain individuals. The situation is further complicated
by limitations in influenza vaccine production, for instance, long production period, limited vaccine capacity and lack
of cross-protection against various influenza A virus strains. To solve these issues, development of universal influenza
vaccine based on conserved antigens such as non-stuctural protein 1 (NS1) has been endeavoured. NS1 protein is highly
conserved in all influenza A virus strains known by far, produced abundantly on infected cell surfaces and responsible for
maintaining virulence. Furthermore, cytotoxic T-lymphocytes that are active against NS1 were also reported to be able
to avoid shedding of influenza in hosts. To better inhibit influenza infections, oral immunization has long been proposed
due to feasibility of this method to be implemented and safer for recipients while able to target influenza A viruses from
the entry point. Lactobacillus has been vastly studied for its roles as bacterial carrier in oral vaccine development due
to its significant probiotic properties. For examples, stimulation of immune responses in oral and airway mucosal layers,
high colonization in oral and airway mucosal layers and great natural adjuvant effects. In this light, influenza universal
oral vaccine developed using NS1 dan Lactobacillus should be further studied in influenza oral vaccine design.
A study was conducted to determine the prevalence of fungal infection in cancer patients warded at the Hospital Kuala Lumpur. There were 138 patients included in the study: 74 with haematological malignancies and 64 with solid tumours. Serological results showed that for candidiasis 16/138 (11.5%) and 10/134 (7.5%) were greater than 1:8 reactive for antigen and antibodies respectively. For aspergillosis, 29/122 (23.8%) sera were antigen reactive. Only 1/133 each (O.SO/o) was reactive for cryptococcal antigen and antibody. The types of malignancies, the age of the patient, a history of neutropenia, the duration of the cancer, the number of courses of chemotherapy or radiotherapy were not predictive of fungal infections. In view of the high serological evidence of fungal infection and the lack of a reliable diagnostic test, empirical antifungal treatment must be considered in all febrile neutropenic cancer patients.
Hidroksiapatit (HA) adalah sejenis kalsium fosfat yang merupakan komposisi kepada kebanyakan fasa mineral tulang dan enamel gigi. HA bersifat bioserasi dan berkonduksi osteo selain mempunyai afiniti biologi dan kimia yang bagus untuk tisu tulang. Dengan ciri tersebut, HA diguna secara luas sebagai graf tulang dan bahan salut bagi implan tisu keras manusia. Walau bagaimanapun, kerapuhan dan keliatan patah yang rendah HA tersinter menghadkan penggunaannya dalam aplikasi bebanan yang tinggi. Kajian ini tertumpu kepada mengenal pasti kesan atmosfera sinteran dengan gas nitrogen (N2) ke atas sifat mekanik HA untuk aplikasi pergigian. Serbuk nano HA dicirikan dengan menggunakan mikroskop elektron pancaran. Cakera silinder HA dihasilkan dengan kaedah penekanan ekapaksi. Kemudian, cakera silinder HA dikenakan tekanan isostatik sejuk dan disinter dalam dua atmosfera sinteran yang berbeza iaitu sinteran dalam gas N2 dan sinteran dalam udara pada suhu 1300°C. Ketumpatan, mikrostruktur, kestabilan fasa dan mikrokekerasan HA tersinter dicirikan. Secara keseluruhan, sinteran dengan menggunakan gas N2 menyebabkan pertumbuhan saiz butiran yang lebih besar dengan ketumpatan relatif dan mikrokekerasan yang lebih tinggi jika dibandingkan dengan atmosfera sinteran dalam udara. Dalam kajian ini, HA yang disinter dengan menggunakan gas N2 pada suhu 1300°C menunjukkan mikrostruktur yang lebih tumpat, ketumpatan relatif (94%) dan mikrokekerasan (4.07 GPa) yang lebih tinggi jika berbanding dengan sinteran dalam udara tanpa penguraian HA. Kesimpulannya, penggunaan atmosfera sinteran dengan menggunakan gas N2 pada suhu 1300°C dapat meningkatkan sifat kekerasan Vickers nanokomposit HA dengan mikrostruktur yang padat.
The Nipah virus is highly virulent to swine and humans. The envelope attachment glycoprotein (G) of Nipah virus
plays a key role in viral entry and induction of neutralizing antibody in mammalian hosts, thus is considered a good
candidate for vaccine development. Plant transient expression systems are gaining recognition as a viable alternative
for the production of vaccine antigens. In this study, we expressed the Nipah virus G protein heterologously in Nicotiana
benthamiana using an agroinfiltration approach. The highest expression of recombinant G protein in N. benthamiana at
RNA and protein levels was detected on day 9 post-infiltration. Western blot analysis demonstrated that the purified G
protein reacted specifically with rabbit anti-Nipah Virus serum, indicating its potential for vaccine use.
In general, demand feeding devices are equipped with a mechanical trigger switch. Such a switch is not suitable for
juvenile fish with a small body size, because the body weight is insufficient to trigger the feeder. An infrared light
sensor that does not require the fish to push a feeder switch is more suitable for small fish. The brown-marbled grouper
Epinephelus fuscoguttatus is an important fish species in Southeast Asia. The purpose of this study was to compare the
growth rates (GRs) of brown-marbled grouper juveniles reared using customised demand feeding devices with an infrared
light sensor (the infrared light demand feeder (IRDF) group) and automatic feeding devices (the automatic feeder (AF)
group). The results indicated that GRs of standard lengths and body weights showed no significant differences using
one-way analysis of variance; however, the standard length of the IRDF group showed a tendency of a higher GR than
the AF group. Although the feed conversion ratio (FCR) also showed no significant difference, the FCR of the IRDF group
was more efficient, indicating that the IRDF group yielded a more desirable FCR. These results indicate that IRDF can be
used in the culture of brown-marbled grouper juveniles. In view of the working schedule of the fish farm staff, IRDF are
superior to other feeding devices, because they are less labour-intensive than usual tasks. We conclude that IRDF is a
useful feeding system for aquaculture.
Metallic foams are a new class of materials that have a great potential to be used in various functional and structural applications. Due to their competitive price compared to aluminium, metallic foams are anticipated to become an alternative material for light-weight structures. In this study, stainless steel foams are fabricated using a powder space holder method. The materials used include stainless steel powder, a novel space holder glycine and binders consisting of palm stearin and of polyethylene (PE). The stainless steel foams are sintered at 1100o C, 1200o C and 1300o C with sintering times of 1, 2 and 3 h, respectively, to investigate the effects of the sintering parameters on the compressive yield strength of the stainless steel foams. The results showed that all of the stainless steel foams produced exhibit the general behaviours of metal foams. The sintering time is the most significant parameter that influences the compressive yield strength of stainless steel foams. Increasing the sintering temperature and sintering time will increase the compressive yield strength. The interaction between the sintering temperature and sintering time is found to be not statistically significant.
The ratios of Tween 80 to Span 80 are important in controlling the physicochemical characteristics of curcumin niosomes. Thus, in this study the correlations of combination of Tween 80 and Span 80 at different ratios with the size and stability of curcumin niosomes were investigated. Initially curcumin was dissolved in chloroform and methanol (v/v= 3:1), followed by the preparation of niosomes by thin-film hydration method with the mole ratio of mixture of Tween 80 and Span 80 to cholesterol of 2:1. The ratios of Tween 80 to Span 80 in the mixture were set at 1:1, 1:9, 9:1, 2:3 and 3:2 with Hydrophilic-Lipophilic Balance (HLB) values of 9.65, 5.37, 13.93, 8.58 and 10.72, respectively. As expected, niosome barely formed at HLB value 13.93. This might be due to the molecule being hydrophilic and well hydrated which inhibits the formation of a stable bilayer in solution. It was found that the formulation which has higher Span 80 and lower HLB value leads to larger niosome before sonication. The largest niosome size achieved from Tween 80 to Span 80 ratio of 1:9 and 2:3 was 77.36 and 59.85 μm, respectively. On the other hand, for the ratio of 3:2, the largest niosome formed before sonication was less than 50.00 μm. After sonication, the niosome size reduced to less than 35.00 μm for all formulations. It was showed that formulation with Tween 80 to Span 80 ratio of 1:9 has superior curcumin entrapment efficiency and best stability during storage. The results demonstrated that appropriate ratio of Tween 80 to Span 80 was necessary for forming small and stable curcumin niosomes.
The objective of the present study was to determine the total phenolic content (TPC), total flavonoid content (TFC) and
antioxidant properties, i.e. 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and ferric reducing
antioxidant potential (FRAP) assay, of red- and yellow-fleshed watermelon rind powders prepared using different drying
conditions (hot-air oven drying at 40 and 60°C and freeze drying). All the samples were subjected to four different
solvent extract using water, methanol, ethanol and acetone prior analyses. Water extract from red- and yellow-fleshed
watermelon rind powders presented highest value for TPC and TFC. However, methanol extract samples showed highest
value for antioxidant properties (DPPH and FRAP) followed by acetone, ethanol and water extract. By comparing the
drying conditions, all samples dried using hot-air dryer at 40 and 60°C had significantly higher (p<0.05) in TPC value
than the samples dried using freeze dryer. However, samples dried using freeze dryer showed highest in DPPH and
FRAP values. The present obtained results would be useful to the food and pharmaceutical industries for developing of
functional ingredients.
Problem-based learning (PBL) i s an educational reform that is now becoming a household word in higher education, particularly in medical schools. Many medical schools have implemented a full problem-based learning curriculum (PBLC) whiIe some have included PBL into selected units of the course in an otherwise conventional cumculum (embedded PBL) and others run their tutorials in a PBL manner within a modified conventional curriculum (hybrid curriculum). Yet there are others who claim that small components of PBL in a conventional curriculum are not PBL at all. Thus amateurs in the subject matter find difficulty in evaluating the logistics and outcome of these variations. This article focuses or, the general characteristics of PBL and how this learning method can help enhance independent learning and critical thinking, whether in a full, embedded or hybrid curriculum. The extent of PBL to be included and which of the three types is to be adopted depends on the objective of the undergraduate medical course as determined by the faculty, resources available, limitations, feedback on the existing curriculum and various other factors. KEYWORDS: Problem-based Learning (PBL); Embedded PBL; Hybrid PBL; New Integrated Curriculum (NIC).
RNA-seq has become an essential tool in molecular research. Nevertheless, application of RNA-seq was limited by cost and technical difficulties. Illumina has introduced the cost effective and ease to handle Truseq Targeted RNA Sequencing. In this study, we present the requirements and the optimization procedure for this Truseq Targeted RNA sequencing on cell line. Total RNA was recommended as starting materials but it required optimization including additional purification step and adjusting the AMPure beads ratio to eliminate unwanted contaminants. This can be resolved by using PolyA-enriched mRNA as starting material. TREx is a useful assay to evaluate gene expression. Quality library of TREx can be prepared by adding multiple washing steps or changing input sample to mRNA.
Amphibians have undergone catastrophic declines worldwide and the major histocompatibility complex (MHC) is an
attractive candidate for investigating the link between adaptive variation and individual fitness. Nonetheless, little research
has been conducted to study the amphibians MHC genes. Thus, MHC class Ia of Odorrana margaretae, a representative
species of the Odorrana, was isolated and used to analyze the adaptive mechanism. We isolated alleles and determined
evolution patterns of MHC class Ia in O. margaretae. The phylogenetic tree of MHC class Ia was reconstructed to
understand the evolutionary relationship of Anura. In O. margaretae of MHC class Ia: there were more non-synonymous
mutations compared to synonymous mutations; there are at least two loci; the recombination alleles took place for a
large proportion (55.56%) of the alleles and recombination usually took place between whole exon of Ia genes of MHC
class; positive selection sites were detected and most (10 of 19) located at the α1 and α2 domains belonged to the
presumed ABS. The diversity of genes of MHC class Ia was led by recombination, gene duplication and positive selection.
Trans-species polymorphisms of anuran genes of MHC class Ia were evident in anurans. Therefore, the MHC class Ia
present demonstrative candidates for investigating the link between adaptive variation and individual fitness. We argue
that knowledge of the MHC of other anuran lineage, especially focusing a genus such as O. margaretae, should provide
an overall more complete picture of the organization of MHC in anurans.
Biosensor optik berasaskan hemoglobin (Hb) terpegun pada mikrosfera poli(n-butil akrilat-co-N-akriloksisuksinimida)
[poli(nBA-NAS)] telah dibangunkan bagi mengesan kepekatan ion nitrit (NO2
-
). Kompleks HEM ferum dalam Hb
memangkinkan tindak balas penurunan ion NO2
-
kepada nitrik oksida (NO) lalu bergabung dengan deoksihemoglobin
(HbFe2+) membentuk kompleks ferum-nitrosil-hemoglobin (HbFe2+-NO) yang berwarna hijau kekuningan. Spektrofotometer
pantulan gentian optik digunakan untuk memantau kepekatan ion NO2
-
secara kuantitatif berdasarkan perubahan warna
Hb terpegun pada mikrosfera poliakrilat daripada perang kemerahan ke hijau kekuningan pada panjang gelombang
pantulan maksimum 668 nm. Pencirian terhadap biosensor nitrit reflektometrik melibatkan ujian kesan pH, kesan
kepekatan Hb, julat rangsangan linear, kebolehasilan, jangka hayat dan kesan gangguan ion telah dijalankan. Biosensor
ion NO2
- optik terbangun memaparkan julat linear dinamik daripada 5 hingga 50 mg mL-1 (R2
=0.9894) pada pH7.0 dengan
had pengesanan (LOD) sebanyak 3.3 mg mL-1 dan nilai sisihan piawai relatif (RSD) kebolehasilan biosensor sebanyak
5.8%. Jangka hayat biosensor optik nitrit tersebut adalah selama 36 hari dan majoriti ion asing yang sering wujud
bersama ion NO2
- dalam sampel air semula jadi tidak menunjukkan kesan gangguan yang bererti terhadap penentuan
ion NO2
-
menggunakan biosensor optik terbangun kecuali ion Hg2+, Ag+, Br-
dan S2-.
Introduction: Negative automatic thoughts refer to subconscious thinking patterns in response to irrational and self-defeating stimuli. Individuals who are depressed tend to have biased negative automatic thoughts. Therefore, there is a need to refine our understanding of what contributes to the development of adolescent depressive symp- toms. The current study examined the mediating role of negative automatic thoughts in the link between maternal attachment and depressive symptoms among late adolescents in Pakistan. Methods: This cross-sectional study fo- cused on 936 (males=423, females=513) students aged 17 to 19 years old, who were selected through multistage cluster sampling technique. Respondents answered self-report questionnaires which assessed maternal attachment, depressive symptoms, and negative automatic thoughts. Structural equation modelling was utilised to analyse the relationships among the research variables. Results: The findings indicated that 39.3% had negative automatic thoughts; whereas, 5.6% experienced severe depressive symptoms. Correlational analysis showed that maternal attachment had a negative association with depressive symptoms and negative automatic thoughts. In contrast, de- pressive symptoms were positively related to negative automatic thoughts. Structural analysis revealed that negative automatic thoughts partially mediated the association of maternal attachment with depressive symptoms among late adolescents. Findings specifically demonstrated that those with insecure maternal attachment tend to have negative automatic thoughts, which subsequently lead to depressive symptoms. Conclusion: In conclusion, findings highlight the importance of secure attachment to the mother and positive automatic thoughts in combating depressive symp- toms; thus ensuring a healthy psychological development for adolescents.
The aim of this systematic review was to evaluate the effectiveness of educational programs on parenting stress and coping mechanism among parents of children with Autism Spectrum Disorder. Our current review retrieved the articles from databases such as CINAHL, Springer, Ovid, PubMed, Google Scholar, and EBSCO host. Only articles published between the years of 2000 and 2018 in these databases were recruited using keywords such as Autism Spectrum Disorder, education program, parenting stress, coping mechanism, and coping strategies. The search gen- erated 17 articles; 8 articles were relevant. This systematic review provides an important opportunity to advance our understanding of the effectiveness of the educational program for reducing parenting stress and improving coping mechanism among parents of children with Autism Spectrum Disorder. Nurses could also have a pivotal role in delivering the educational program for parents of children with ASD.
Introduction: CRISPR/Cas9 nuclease has gained popularity as a genome editing tool due to its straight-forward mechanism. However, there are concerns that CRISPR nuclease would cause off-target and toxicity. The CRISPR/ Cas9 D10A nickase was designed to enhance genome editing. Nevertheless, this raised the question of whether the efficiency of nickase is compromised compared to CRISPR/Cas9 nuclease. Targeting HIV genes, we investigated if CRISPR nuclease performed better than the nickase in efficacy and safety. Methods: CRISPR nucleases and nickases were designed to target Gag, Pol, Rev, Vif, Tat and LTR. HIV latently infected cell line, ACH-2, was transfected with the nucleases and nickases. Changes to viral load after CRISPR treatment was measured using p24 ELISA. Safety of nuclease and nickase was monitored using GFP expression with fluorescence microscopy and flow cytometry. Targeting two sites within the same gene, and targeting multiple genes concurrently were also studied to determine efficacy of CRISPR in reducing viral load. Results: A 44.9 to 68.1% and a 34.4 to 49.7% decrease in viral load was seen in CRISPR nuclease and nickase respectively. Microscopy and flow cytometry results showed that the nickase system was slightly toxic with a 0.31 to 0.7-fold cell death. There was a 34% decrease in viral load when two sites were targeted within a gene, and the largest decrease was seen when all the nucleases were combined, giving a 75.4% decrease in viral load at day 5. Conclusion: The knowledge gained from this study will be employed to im- prove genome editing in other disease models.