Screening for Acyl Carrier Protein (ACP) Gene of Oil Palm (Elaeis Quineensia) Using a Synthetic ACP DNA Probe

Ac DNA library was con structed inphagemid vector pT7T3 using messenger RNA (mRNA) extracted from IS - weekoil palm me socarp as template. The cDNA inserts were detected from agarose gel analysis and ranged in length from 130 to 1300 base pairs.This library was screened with a synthetic ACP probe.Of the 2,000 clones screened using colony hybridisation analysis, 550 clones which gave high intensity of signal were then further screened using phagemid blot hybridisation analysis. This met hodyielded 184 clones which showed a high intensity of signal. The phagemids from the seclones were then screened by Southern hybridisation analysis using a synthetic ACP DNA probe

Gene discovery via expressed sequence tags from oil palm (Elaeis guineensis Jacq.) mesocarp.

Expressed sequence tags (ESTs) have been used for many applications such as to reveal gene expression patterns, gene regulation and sequence diversity. A total of 1011 unique transcripts corresponding to 1463 genes have been identified from the ESTs generated from 17-week oil palm mesocarp cDNA library. This approach was found to be successful in the discovery of new and important genes expressed in the mesocarp tissue which are associated with the various cellular processes of the tissue. It was observed that 12.9% of the total genes expressed in the 17-week mesocarp cDNA library can be categorized under metabolism. This is in agreement with the function of the tissue which is involved in many biochemical processes including amino acid and fatty acid metabolism. Most importantly are the discoveries of genes playing important roles in the fatty acid and wax biosynthesis pathway such as acetyl-CoA carboxylase, stearoyl-ACP desaturase, acyl carrier protein (ACP), lysophosphatidic acid acyltransferase, ?6-palmitoyl-ACP desaturase and lipase. These genes can serve as targets for genetic manipulation where such endeavours have been extensively carried out in other plants such as Brassica napus and Olea europaea to help increase the economic value of the oil. Genes and protein associated with ethylene synthesis and signal transduction pathway were also identified from the 17-week mesocarp ESTs. Dot blot analysis was carried out to help in identifying potential tissuespecific genes, which can lead to the isolation of the tissue-specific promoters for manipulation of the mesocarp tissue. This is in particular to direct accumulation of transgenic products, such as new specialty oils and value-added products like pharmaceuticals and nutraceuticals to the mesocarp

Differentially expressed transcripts related to height in oil palm

A subtractive cDNA library was constructed using MPOB Planting Series 1 (PS1) population to isolate differentially expressed transcripts associated with height increment in oil palm. After differential screening, 98 clones were identified to be potentially positive with cDNA inserts ranging from 250 to 1000 bp. A total of 123 sequences generated and low quality sequences ≤20 were eliminated using Phred program to assess the sequence quality and determine the accurate consensus sequence. Contig assembly by CAP3 program generated 51 contigs and four singletons (55 unigenes). Blast search showed 49 unique sequences had significant match to various plant species in the GeneBank database with E-value ≤1e-5, and four sequences showed no significant similarity. Gene ontology analysis output from Blast2 GO program revealed that the sequences encoding for auxin responsive protein, circadian clock-associated protein1, zinc-finger protein and basic leucine zipper protein were potentially associated to dwarfism. The genes were identified based on their putative functions in regulating height in plants, particularly in growth hormone biosynthesis such as auxin (Aux/IAA), gibberellins (GA) and brassinosteroids (BR)

Efficient protocol improved the yield and viability of oil palm protoplasts isolated from in vitro leaf and mesocarp

The absence of a cell wall on the protoplast contributes to its versatility. Its flexibility for DNA manipulation and the possibility of rapid cell-based assay is desirable in the plant biotechnology field. This study was carried out to improve protoplast isolation from oil palm in vitro leaf and mesocarp tissues. The factors affecting protoplast isolation efficiency were optimized, including the protocols and enzyme composition involved, focusing on the oil palm in vitro leaf first. Incubation of oil palm leaf sample with an enzyme mixture of cellulase R-10, macerozyme R-10, driselase, and pectolyase Y-23, for 14 h has successfully produced up to 2.5 × 106 protoplasts g-1 fresh weight (FW)-1 with 95% viability. Incubation of oil palm mesocarp tissue with the optimized enzyme mixture for 2 h at static condition has also successfully produced 3.98 × 106 protoplasts g-1 FW-1 with 85% viability. Besides, it was found that increasing the sample's surface area in contact with enzyme solution by slicing the samples into narrow strips and thin layers has improved the penetration of enzymes into the tissues and enhanced the isolation efficiency. In addition, a plasmolysis step before enzymatic treatment has also improved the protoplast viability by minimizing the damage incurred during isolation. The successful isolation of protoplast from oil palm leaf and mesocarp has enabled the study of gene function and the characterization of endogenous tissue-specific promoter being carried out in vivo

Technical Aspects and Clinical Limitations of Sperm DNA Fragmentation Testing in Male Infertility: A Global Survey, Current Guidelines, and Expert Recommendations

PURPOSE: Sperm DNA fragmentation (SDF) is a functional sperm abnormality that can impact reproductive potential, for which four assays have been described in the recently published sixth edition of the WHO laboratory manual for the examination and processing of human semen. The purpose of this study was to examine the global practices related to the use of SDF assays and investigate the barriers and limitations that clinicians face in incorporating these tests into their practice. MATERIALS AND METHODS: Clinicians managing male infertility were invited to complete an online survey on practices related to SDF diagnostic and treatment approaches. Their responses related to the technical aspects of SDF testing, current professional society guidelines, and the literature were used to generate expert recommendations via the Delphi method. Finally, challenges related to SDF that the clinicians encounter in their daily practice were captured. RESULTS: The survey was completed by 436 reproductive clinicians. Overall, terminal deoxynucleotidyl transferase deoxyuridine triphosphate Nick-End Labeling (TUNEL) is the most commonly used assay chosen by 28.6%, followed by the sperm chromatin structure assay (24.1%), and the sperm chromatin dispersion (19.1%). The choice of the assay was largely influenced by availability (70% of respondents). A threshold of 30% was the most selected cut-off value for elevated SDF by 33.7% of clinicians. Of respondents, 53.6% recommend SDF testing after 3 to 5 days of abstinence. Although 75.3% believe SDF testing can provide an explanation for many unknown causes of infertility, the main limiting factors selected by respondents are a lack of professional society guideline recommendations (62.7%) and an absence of globally accepted references for SDF interpretation (50.3%). CONCLUSIONS: This study represents the largest global survey on the technical aspects of SDF testing as well as the barriers encountered by clinicians. Unified global recommendations regarding clinician implementation and standard laboratory interpretation of SDF testing are crucial

Controversy and Consensus on Indications for Sperm DNA Fragmentation Testing in Male Infertility: A Global Survey, Current Guidelines, and Expert Recommendations

Purpose: Sperm DNA fragmentation (SDF) testing was recently added to the sixth edition of the World Health Organization laboratory manual for the examination and processing of human semen. Many conditions and risk factors have been associated with elevated SDF; therefore, it is important to identify the population of infertile men who might benefit from this test. The purpose of this study was to investigate global practices related to indications for SDF testing, compare the relevant professional society guideline recommendations, and provide expert recommendations. Materials and Methods: Clinicians managing male infertility were invited to take part in a global online survey on SDF clinical practices. This was conducted following the CHERRIES checklist criteria. The responses were compared to professional society guideline recommendations related to SDF and the appropriate available evidence. Expert recommendations on indications for SDF testing were then formulated, and the Delphi method was used to reach consensus. Results: The survey was completed by 436 experts from 55 countries. Almost 75% of respondents test for SDF in all or some men with unexplained or idiopathic infertility, 39% order it routinely in the work-up of recurrent pregnancy loss (RPL), and 62.2% investigate SDF in smokers. While 47% of reproductive urologists test SDF to support the decision for varicocele repair surgery when conventional semen parameters are normal, significantly fewer general urologists (23%; p=0.008) do the same. Nearly 70% would assess SDF before assisted reproductive technologies (ART), either always or for certain conditions. Recurrent ART failure is a common indication for SDF testing. Very few society recommendations were found regarding SDF testing. Conclusions: This article presents the largest global survey on the indications for SDF testing in infertile men, and demonstrates diverse practices. Furthermore, it highlights the paucity of professional society guideline recommendations. Expert recommendations are proposed to help guide clinicians

Technical aspects and clinical limitations of sperm DNA fragmentation testing in male infertility: A global survey, current guidelines, and expert recommendations

Purpose Sperm DNA fragmentation (SDF) is a functional sperm abnormality that can impact reproductive potential, for which four assays have been described in the recently published sixth edition of the WHO laboratory manual for the examination and processing of human semen. The purpose of this study was to examine the global practices related to the use of SDF assays and investigate the barriers and limitations that clinicians face in incorporating these tests into their practice. Materials and Methods Clinicians managing male infertility were invited to complete an online survey on practices related to SDF diagnostic and treatment approaches. Their responses related to the technical aspects of SDF testing, current professional society guidelines, and the literature were used to generate expert recommendations via the Delphi method. Finally, challenges related to SDF that the clinicians encounter in their daily practice were captured. Results The survey was completed by 436 reproductive clinicians. Overall, terminal deoxynucleotidyl transferase deoxyuridine triphosphate Nick-End Labeling (TUNEL) is the most commonly used assay chosen by 28.6%, followed by the sperm chromatin structure assay (24.1%), and the sperm chromatin dispersion (19.1%). The choice of the assay was largely influenced by availability (70% of respondents). A threshold of 30% was the most selected cut-off value for elevated SDF by 33.7% of clinicians. Of respondents, 53.6% recommend SDF testing after 3 to 5 days of abstinence. Although 75.3% believe SDF testing can provide an explanation for many unknown causes of infertility, the main limiting factors selected by respondents are a lack of professional society guideline recommendations (62.7%) and an absence of globally accepted references for SDF interpretation (50.3%). Conclusions This study represents the largest global survey on the technical aspects of SDF testing as well as the barriers encountered by clinicians. Unified global recommendations regarding clinician implementation and standard laboratory interpretation of SDF testing are crucial

Controversy and consensus on the management of elevated sperm DNA fragmentation in male infertility: A global survey, current guidelines, and expert recommendations

Purpose Sperm DNA fragmentation (SDF) has been associated with male infertility and poor outcomes of assisted reproductive technology (ART). The purpose of this study was to investigate global practices related to the management of elevated SDF in infertile men, summarize the relevant professional society recommendations, and provide expert recommendations for managing this condition. Materials and Methods An online global survey on clinical practices related to SDF was disseminated to reproductive clinicians, according to the CHERRIES checklist criteria. Management protocols for various conditions associated with SDF were captured and compared to the relevant recommendations in professional society guidelines and the appropriate available evidence. Expert recommendations and consensus on the management of infertile men with elevated SDF were then formulated and adapted using the Delphi method. Results A total of 436 experts from 55 different countries submitted responses. As an initial approach, 79.1% of reproductive experts recommend lifestyle modifications for infertile men with elevated SDF, and 76.9% prescribe empiric antioxidants. Regarding antioxidant duration, 39.3% recommend 4–6 months and 38.1% recommend 3 months. For men with unexplained or idiopathic infertility, and couples experiencing recurrent miscarriages associated with elevated SDF, most respondents refer to ART 6 months after failure of conservative and empiric medical management. Infertile men with clinical varicocele, normal conventional semen parameters, and elevated SDF are offered varicocele repair immediately after diagnosis by 31.4%, and after failure of antioxidants and conservative measures by 40.9%. Sperm selection techniques and testicular sperm extraction are also management options for couples undergoing ART. For most questions, heterogenous practices were demonstrated. Conclusions This paper presents the results of a large global survey on the management of infertile men with elevated SDF and reveals a lack of consensus among clinicians. Furthermore, it demonstrates the scarcity of professional society guidelines in this regard and attempts to highlight the relevant evidence. Expert recommendations are proposed to help guide clinicians

Carbon Based Electrochemical Capacitor in 1M H2SO4 Electrolyte

In this article, the EC electrode is mainly consists of AC (97%) and graphene (3%) was added to enhance the electrode’s conductivity. Graphene not only has unique physical and chemical properties but also provides a great opportunity to understand the fundamental chemical and physical processes involved in the complex systems [Candelaria, 2012]. The electrode fabrication starts with the slurry preparation. The material is weighing by using weight balance to produce a 20 g electrode. The materials used are AC, graphene, KS6, binder, and 60 g NMP. The viscosity of the slurry needed is ca. 10,000 MPa.s with 50% solid content. The mixing process is repeated if the viscosity does not achieve the general standard. 1M sulfuric acid (H2SO4) was used as electrolyte, and the CV and charge-discharge measurements were conducted in 0.0 – 1.0 V potential range by using Basic DC Voltammetry System

Green magnetic composite sheet from durian shell and nano magnetite particles

Abstract. Biomass-derived materials such as kenaf pulp and wood chips are a perfect candidate to produce magnetic paper. Furthermore, by using biomass waste, such as paddy straw, sugarcane, bagasse and durian shell, the cost of producing magnetic paper can be further reduced while giving added value to the waste. This paper investigates the potential of producing magnetic sheet from durian shell. Initially, durian shells were dried before undergoing the pulping process. The resulted sheet was then combined with magnetic particles, the nano magnetite using either lumen loading or in-situ co-precipitation to produce a magnetic composite sheet. After being loaded with magnetic particles, the composite sheets were tested in terms of the homogeneity of the magnetic particles in the samples, degree of loading of the magnetic particles and the magnetic properties of the samples. Results obtained show a great success in producing the magnetic sheet from durian shell waste and nano magnetite particles. It was also found that the lumen loading method gives better magnetic properties compared to the in-situ co-precipitation method