Assembly of Functional Cellulolytic Enzymes

Gathering of Functional Cellulolytic Enzymes In the current investigation, we detailed the get together of utilitarian cellulolytic proteins utilizing a manufactured, cell-surface designed yeast consortium. Trichoderma reesei endoglucanase II (EGII) and cellobiohydrolase II (CBHII) and Aspergillus aculeatus ÃŽ ²-glucosidase I (BGLI) were shown as combination proteins with the AGA2p C-end of an agglutinin on the cell surface of the diploid yeast strain Saccharomyces. cerevisiae Y5. The immobilization of every compound on the cell surface was affirmed by immunofluorescence microscopy. This sort of yeast consortium permitted helpful advancement of ethanol creation by altering the mix proportions of every phone type for instigating collaboration in cellulose hydrolysis. Next, the immediate ethanol aging from steam-detonated corn stover was explored. The enhanced cellulase-showing consortium delivered 20.4 g/l ethanol from 48.4 g cellulose per liter after 72 h within the sight of a limited quantity of cellulase reagent (0.9 FPU/ml). These discoveries proposed the practicality of the cellulase-showing yeast consortium for concurrent saccharification and aging. Right now, numerous innovative obstructions exist as for the affordable creation of ethanol from lignocellulosic biomasses [1]. During the time spent hydrolyzing cellulose into dissolvable sugars, numerous cellulases including endoglucanase (EG), cellobiohydrolase (CBH), and ÃŽ ²-glucosidase (BGL) are required [2]. United bioprocessing (CBP), which consolidates catalyst creation, hydrolysis, and aging in one stage, is a promising system for successful ethanol creation from lignocellulosic materials. Saccharomyces cerevisiae is the conventional microorganism utilized for ethanol creation, however it can't use cellulosic materials and a saccharification procedure is required before maturation to deliver glucose [3-4]. Various endeavors have been made to design S. cerevisiae strains to communicate cellulases by cell surface building for direct ethanol creation from cellulose, and albeit different bifunctional or trifunctional cellulose-corrupting strains have been developed, the product ivity of cellulose debasement has not been adequately improved [5-9]. No doubt co-articulation of all cellulolytic chemicals in a solitary cell brought about moderately low articulation levels of cellulases, which may have been because of the overwhelming metabolic weight and potential sticking of the discharge hardware [6,7,10]. In this manner, in this examination, we adjusted another system of performing synchronous saccharification and aging with an artificially designed yeast consortium having the ideal properties of cellulolytic capacity and ethanol creation to decrease the metabolic weight. The improvement of a diploid yeast strain is another promising technique for improving articulation levels of heterologous qualities and upgrading the maturation execution of S. cerevisiae. Since diploid strains have better development capacity just as stress resistances contrasted and haploid strains, they are especially appropriate for modern applications. Beforehand, our gathering wrote about the development of a à Ã¢ °-agglutinin articulation framework for hereditary immobilization ÃŽ ²-glucosidase I on the phone surface of S. cerevisiae Y5 (Patent No: ZL200810222897.7, CGMCC2660). This diploid strong yeast strain had numerous focal points, for example, higher ethanol yield, higher protection from ethanol, and higher physiological resistance to inhibitors present in lignocellulosic hydrolysates. Here, we report on our endeavors to show the gathering of practical cellulolytic compounds utilizing an engineered yeast consortium. In this examination, we showed the plausibility of building a novel cell surface designed diploid yeast consortium for direct ethanol creation from phosphoric corrosive swollen cellulose (PASC) and steam-detonated corn stover (CS), a significant advance toward direct ethanol creation from insoluble cellulosic materials. The strains and plasmids utilized in this examination are summed up in Table S1. Saccharomyces cerevisiae Y5 utilized for the yeast cell surface showcase of the cellulolytic catalysts was a recently evolved diploid strain in our lab. E. coli Top 10 was utilized as the host strain for recombinant DNA control. T. reesei was bought from CICC (China Center of Industrial Culture Collection). E. coli transformants were developed in Luria-Bertani medium (1% tryptone, 0.5% yeast extricate and 1% NaCl, pH 7.0) enhanced with 100 ug/ml of ampicillin. S. cerevisiae Y5 transformants were chosen and kept up on Geneticin plates (1% yeast separate, 2% peptone and 2% glucose enhanced with 600 ug/ml Geneticin) at 30 °C , were actuated in YPG (1% yeast remove, 2% peptone, and 2% galactose) at 20 °C. The aging medium was made out of 10 g/l yeast separate, 20 g/l polypeptone and 10 g/l PASC as the sole carbon source. The à ¯Ã¢ ¬Ã¢ lamentous parasite T. reesei was refined in potato dextrose agar med ium (2% potato separate, 2% glucose) at 27 °C. The cDNA was combined from mRNA by utilizing the First-Strand cDNA blend pack (Fermentas). Except if in any case showed, all synthetic substances, media parts and enhancements were of diagnostic evaluation standard and gotten from Sigma-Aldrich (St. Louis, MO, USA). All limitation chemicals were bought from New England BioLabs (Ltd. Beijing). Preliminaries utilized for plasmid development are given in Table S2. Plasmid pAGA1 for over-articulation of the AGA1 quality and plasmid pBGLI for cell surface showcase BGLI were built already [11]. Plasmid pEGII for cell surface articulation of the EGII (egl2) was built as follows. The 1194 bp DNA part encoding the egl2 quality without its local discharge signal was ampli㠯⠬⠁ed with the à ¯Ã¢ ¬Ã¢ rst-strand cDNA arranged from T. reesei as the layout utilizing preliminary sets egl2-For/Rev, this DNA section was brought into the yeast show vector pYD1(Invitrogen) with Kpn I/BamH I. Tangle eliminator was intensified from pYD1 by utilizing groundwork sets MAT-For/Rev and afterward processed with BamH I/EcoR I to make plasmid pYD1-egl2MAT. The KanR piece was gotten from plasmid YIP5-KanR by two-advance cloning. In the first place, the DNA part containing ADH advertiser and KanR ORF was enhanced from YIP5-KanR by PCR utilizing the KanR-For/Rev groundworks and embedded into EcoR I/Apa I site of plasmid pYD1-egl2MAT; next, the ADH eliminator processed with Bgl II/Nde I was additionally brought into pYD1-egl2MAT. The subsequent plasmid was named pEGII. For showing the T. reese i CBHII quality (cbh2) in S. cerevisiae Y5, plasmid pCBHII was made. A 1344 bp quality section coding for the develop area of the CBHII was ampli㠯⠬⠁ed utilizing preliminaries cbh2-For/Rev-KT and brought into plasmid pEGII processed with Kpn I/BamH I for supplanting egl2 to shape pCBHII (Figure 1). Change of S. cerevisiae Y5 was completed utilizing the lithium acetic acid derivation strategy [12]. The plasmid pAGA1 was linearized by Apa I for chromosome reconciliation. The plasmid pYD1 was changed into S. cerevisiae Y5 as a negative control. S. cerevisiae Y5 clones changed with various plasmids (strain Y5/pYD1 contained plasmids pAGA1 and pYD1, strain Y5/EGII contained plasmids pAGA1 and pEGII, strain Y5/CBHII contained plasmids pAGA1 and pCBHII) were chosen and kept up on Geneticin(G418) plates. Immunofluorescence microscopy was proceeded as portrayed beforehand [13]. Immunostaining was proceeded as follows. Initiated recombinant yeast cells communicating cellulases were gathered by centrifugation at 6000 rpm for 5 min and washed with phosphate-cushioned saline (PBS). As the essential immunizer, mouse against Xpress label neutralizer (Invitrogen, R910-25) for EGII and CBHII was utilized at weakening paces of 1:1000. As the subsequent immune response, Fuorescein (FITC)- conjugated goat against mouse IgG(H+L) (Jackson, 115-095-003) was utilized at weakening rate 1:200. Cells and the immunizer were brooded at room temperature. In the wake of washing the cellâ€antibody complex with PBS twice, cell limitations of the cellulases were seen under a fluorescence magnifying lens. Yeast strains Y5 and Y5/pYD1were utilized as control. Yeast cells were prompted in YPG vehicle for 48 h at 20 ºC and collected by centrifugation for 5 min at 6000 rpm, washed with refined water. BGLI action of strain Y5/BGLI was estimated utilizing à ¯Ã¢ Ã¢ ²-nitrophenyl-ÃŽ ²-D-glucopyranoside as the substrate as indicated by a formerly depicted strategy [14]. Endoglucanase and cellobiohydrolase exercises were dictated by hydrolysis of carboxymethyl cellulose (CMC) and phosphoric corrosive swollen cellulose (PASC), individually. PASC was set up from Avicel PH-101 (Fluka Chemie GmbH, Buchs, Switzerland) as undefined cellulose. The cell pellet was resuspended in a response blend of 1% CMC or 1% PASC in 50 mM sodium acetic acid derivation cradle (pH 5.0) with the optical thickness at 600 nm acclimated to 1.0. After a response at 50 ºC for 30 min, the exercises were controlled by DNS strategy [15]. One unit of compound movement was characterized as the measure of catalyst discharged 1 ÃŽ ¼mol lessening sugar from the substrate every moment. The capacities of the designed yeast consortium (Y5/EGII + Y5/CBHII + Y5/BGLI) to aging ethanol from PASC and steam-detonated corn stover were researched. The steam-detonated corn stover utilized in this investigation was given by Henan Tian Guan Group Co., Ltd (Nanyang, Henan, China). The crude material was slashed to 2-3 cm size and rewarded in a steam-detonated vessel at 2.0 MPa for 5 min. The pretreated feedstock was dried at room temperature and straightforwardly utilized as a substrate without washing. The dampness substance of the substrate was 8%. The creation of materials was quantitatively investigated following the NREL Laboratory Analytical Procedure NREL/TP-510-42618 (Structural sugars and lignin) (Sluiter et al., 2008)[16], as appeared in Table 3. A compound blend made out of equivalent measures of cellulase (Sigma-Aldrich, St. Louis, MO

Unification Theory Essays (597 words) - Particle Physics

Unification Theory In molecule material science, an endeavor to clarify the entirety of the key powers and their connections between rudimentary particles in single structure was achieved in principle by the g.u.t. by the great unification hypothesis. In connection to material science these powers can be portrayed as fields that intervene associations between isolated or far off articles. These speculations, for example, eltromagnetism and general relativity began to endeavor the unification of hypotheses, anyway they would develop as the key essentials of the g.u.t. Or then again the great unification hypothesis. At sub nuclear levels, these fields are portrayed as quantum field speculations, which began the thoughts of quantum mechanics. In the 1940's the thoughts quantum electrodynamics (QED), the quantum field hypothesis of electromagnetism, turned out to be completely evolved. In QED, charged particles communicate as they discharge and ingest photons (minute parcels of electromagnetic radiation), in impact trading the photons in a round of subatomic get. This hypothesis has become the model for speculations of different powers. During the 1960s and '70s molecule physicists found that issue is made out of two sorts of essential structure square - the principal particles known as quarks and leptons. The quarks are constantly bound together inside bigger noticeable particles, for example, protons and neutrons. They are limited by the short-extend solid power, which overpowers electromagnetism at sub atomic separations. The leptons, which incorporate the electron, don't feel the solid power. Notwithstanding, quarks and leptons both experience a second atomic power, the frail power. This power, which is liable for specific kinds of radioactivity classed together as beta rot, is weak in examination with electromagnetism. While the image of quarks and leptons started to solidify, significant advances prompted the chance of building up a brought together hypothesis. Scholars started to summon the idea of nearby measure invariance, which proposes balances of the fundamental field conditions at each point in existence. Both electromagnetism and general relativity previously included such balances, yet the significant advance was the disclosure that a measure invariant quantum field hypothesis of the feeble power needed to incorporate an extra communication - specifically, the electromagnetic communication. Sheldon Glashow and companions freely proposed a bound together electro frail hypothesis these powers dependent on the trading of four particles: the photon for electromagnetic cooperations, and two charged W particles and a nonpartisan Z molecule for frail cooperations. During the 1970s a comparative quantum field hypothesis for the solid power, called quantum thermodynamics (QCD), was created. In QCD, quarks communicate through the trading of particles called gluons. The point of scientists presently is to find whether the solid power can be brought together with the electro frail power in a terrific bound together hypothesis (GUT). There is proof that the qualities of the various powers fluctuate with vitality so that they merge at high energies. Be that as it may, the energies included are very high, in excess of a million times as incredible as the vitality size of electro feeble unification, which has just been confirmed by numerous tests. Fantastic brought together hypotheses portray the communications of quarks and leptons inside the equivalent hypothetical structure. This offers ascend to the plausibility that quarks can rot to leptons and explicitly that the proton can rot. Early endeavors at a GUT anticipated that the proton's lifetime must be in the area of 1032 years. This forecast has been tried in tests that screen a lot of issue containing on the request for 1032 protons, yet there is no proof that protons rot. On the off chance that they do in reality rot, they should do so with a lifetime more noteworthy than that anticipated by the least complex GUTs. There is likewise proof to propose that the qualities of the powers don't join precisely except if new impacts become an integral factor at higher energies. One such impact could be another evenness called supersymetry, which is a piece of the g.u.t.

Foreign Aid Speech Free Essays

Australia’s remote guide ought to be diminished Today I’m bringing a genuine subject that has Influenced the world and the advancement of nations around us into viewpoint. Remote guide, and how the legislature should Increase It, however first we need to pose the inquiry, â€Å"What Is outside guide? ‘ Simply, It Is the financial, or military chap given by one country to another for motivations behind help and recovery, for monetary adjustment, or for common resistance. The 620 or gathering of 20 is the head discussion for its members’ universal monetary participation and dynamic. We will compose a custom exposition test on Outside Aid Speech or then again any comparative subject just for you Request Now Also, their reality is a higher priority than at any other time on the grounds that the world’s economy is in such a problematic state thus the choices they cause will to be imperative in the advancement of more unfortunate nations. A piece of this that individuals may not perceive is that when the economy is frail, the least fortunate nations endure the most. $1 dollar can help a hundred people in a poor nation beyond what it can help somebody in a rich or created nation. In spite of the fact that we have more grounded proclivity towards our residents and if cash was similarly powerful we would whose our own residents first, when it’s multiple times progressively compelling in affecting lives, at that point we think we’re every individual. The enduring of a mother whose kid Is wiped out and may pass on Is a widespread worth. Furthermore, If a dollar has so much Impact then I should ask you, not as an understudy, however as a kindred human: for what reason would we say we are for the most part still so voracious? If I somehow managed to solicit you what rate from the government financial plan is spent on remote guide, would you be amazed that it is 0. 35%? That likens to $5. 44 billion dollars of our IN (net national pay) in 2012. The administration hopes to arrive at 0. 5% by 2017, however in ongoing media, Julie Bishop, the Minister of outside issues has actualized a top of 5 billion dollars throughout the following two money related years. The guarantee Australia made with the UN alongside numerous other created nations, was a concurred 0. 7% of our nations IN. The 2014 Australian spending plan IS a messed up political decision guarantee to the world’s least fortunate individuals. The Abbott Government has destroyed its guarantee to Increase help. This year’s spending plan, declared two days back, shows that the Australian government Is cutting trench the most defenseless individuals from the networks the world over. Consistently we inhale, 30, 000 youngsters younger than five kick the bucket from preventable infections. It is made apparent that Australia’s commitment to ease destitution through abroad guide is fruitful. The normal future of people living in creating nations has expanded by roughly twenty years in the most recent decade, which is significant rather than the figures fifty years prior. Vaunt has decreased the frequency of intestinal sickness by 80% with the help of Australian guide. It is a legend that rule help doesn’t work. 79% of Australians, as per the fundamental report, concur that the Government ought to lessen spending in outside guide when 41% don’t realize what it Is. The numbness encompassing remote guide Is the thing that overlords enduring of people living In creating nations. Australia Is as of now giving less outside guide contrasted with other created nations. We are positioned 1 thirteenth. The Abbott Government’s nullification of Causal, which deals with our duty dollars for abroad guide, Is on the grounds that they need to see a superior arrangement between help, exchange and conciliatory erection and help programs going in another direction† yet there is no proof for this. Help versus.. Tact is a bogus polarity. Helping neighboring nations create solid economies implies a superior market for our own merchandise. Helping our neighbors improve their wellbeing frameworks implies less wellbeing dangers. Expanding the quantity of kids in our area going to class decreases the open door for teaching instead of training. The achievement of our neighbors are beneficial for us. In 2006 and 2007, Australia gave 2. 9 billion dollars through Said and in 2013, 2014, it developed to 5. 44 lion dollars, nearly multiplying the abroad help. Presently, the extent of help to IN has returned to 2000, 2001 levels under Abbot’s government, particularly since remote guide will develop in accordance with the Consumer Price Index or expansion rate. With the assistance of worldwide activities, the quantity of individuals living in destitution has fallen by 200 million individuals since 1980 and expanding. Australia shouldn’t walk out on the less lucky as a well-to-do nation. The kids enduring each and every day depend on our worldwide guide. The administration has sliced the outside guide spending which asses numerous issues and places a strain on our relationship with different nations. In the event that Australia works with the nations we’re helping, to divert outside guide assets to needs resources, deregulate their economy and opened up their economy to worldwide exchange for monetary development, at that point the yield of remote guide would be powerful in framing a manageable economy and mitigating neediness. We should raise our Foreign guide financial plan to the concurred 0. 7% IN. Australia is a liberal nation, we can stand to give help to those out of luck and develop, as a nation, however as people of this world. Instructions to refer to Foreign Aid Speech, Papers

Define and Evaluate Law of Evidence

Questions: 1.Describe and assess the reasons why an admonition might be given to a jury?2.Describe and talk about the impact of the Evidence Act 2008 on verification warnings?3.Identify and survey the conditions under which an appointed authority may even now give confirmation warning?4.Determine the basics in the planning of the case?5.Identify the significant components of assortment, structures and substance, in the social affair of verification of proof and survey for application?6.Identify the procedure in convincing the creation of proof and assess for application?7.Determine the procedures for offering kinds of proof, for example, reports; photos; maps; and plans; genuine proof, and views?8.Describe and examine the reason and substance of opening and shutting addresses?9.Research and record the historical backdrop of the advancement of Uniform Evidence Legislation?10.Research and talk about the targets and supports for the presentation of the Evidence Act 2008 (Vic)?11.Identify and depic t the structure and arrangements of the Evidence Act 2008 (Vic) and its relationship with the Evidence Act 1995 (Cth) and evaluate for the application to rehearse? Answers: 1. An admonition to the jury board is given in the situations where a charge is made in regards to a rape. This is on the grounds that such cases are horrible in nature for various individual and for shifted reasons[1]. The admonition is given in such cases with the goal that a jury can be made mindful about the impacts which may result on a denounced, because of postponement between the wrongdoing that has been asserted and the trial[2]. 2. The continuing necessity according to which the proof must be substantiated, with the sole exemption of the prevarication cases, the prerequisite has been canceled because of Section 164[3] of the Evidence Act 2008[4]. In this way, a jury isn't required to caution that to follow up on groundless proof is perilous or even give an admonition about the equivalent. Area 164(4) gives that an appointed authority ought not immediate the jury about the supports absence[5]. 3. Area 164(2) states that the arrangements contained in this segment are not pertinent on prevarication offenses or such related offences[6]. Further according to segment 164(5), the adjudicator is required to coordinate the jury that the individual blamed must be seen as liable, when they have been fulfilled with respect to the proof, which can demonstrate the blame and such a proof is corroborated[7]. In any event, when there is a restriction over authenticating bearings, in situations where the jury has been cautioned with respect to the conceivable lack of quality of the observer, and furthermore, there is nearness of proof which can bolster or affirm the proof of the observer, at that point it is considered as fitting for the jury to search for steady proof, as was found on account of R v Connors[8] and R v Milton[9]. 4. The readiness of the case helps in winning a large portion of the fight. Along these lines, the essentials of groundwork for a case are to be followed appropriately before the case is really introduced under the watchful eye of an official courtroom. The as a matter of first importance viewpoint in this is to investigate the quality, just as, the shortcoming of the case. The subsequent stage is to set up the records and the proof for the preliminary. The observers are a key to any case, and they must be appropriately distinguished and arranged heretofore. Finally, the key purposes of the case must be polished again and again again[10]. 5. There are sure vital components while gathering the confirmation of any sort of proof and these identify with its assortment, structure and substance. Proof comprises of a scope of things, and principally has three structures, which are genuine proof, declaration and documents[11]. The genuine proof is gathered to demonstrate charges made in a wrongdoing, and its substance incorporate the assortment of things like tissues, semen, spit, fingerprints and blood[12]. The proof law works over the standards with respect to the verification of realities in criminal, just as, common trials[13]. 6. According to Section 45 of the Evidence Act 2008, an individual can be constrained to deliver a report or such other proof which is accessible to the gathering, under the watchful eye of the court or to some other gathering, through a request for the court[14]. The proof can be constrained to be created uniquely in situations where a gathering has interrogated an observer or is doing so directly, with respect to a former portrayal or a conflicting proclamation, which has been purportedly made by someone else or the observer, individually, and which has been appropriately recorded in a document[15]. The proof that has been created would then be inspected by the court and the bearing in regards to its utilization is likewise given by the court. The court, at its watchfulness can likewise concede such created proof, in any event, when the equivalent has not been offered by the party[16]. 7. The customary law rule, subject to certain exemptions, gives that the first records must be offered in proof with the goal that their substance can be demonstrated. This standard is otherwise called the first report rule, be that as it may, the Uniform Evidence Act in Victoria, through its area 51 annuls this rule[17]. This demonstration gives that the proof can be entered by offering the record under segment 48 of this act[18]. The strategies or procedure of offering the records has been secured under this area. For example, offering a transcript of a chronicle, or a duplicate of an archive, is taken as offering of proof. Segment 53 contains the arrangements with respect to offering of perspectives, in way of exhibition or inspection[19]. 8. The opening and shutting tends to goes about as the chance of tending to the jury straightforwardly regardless, which permits the opportunity to give the jury a comprehension with respect to the job of the gathering for the situation, and the job of the proof that has been introduced under the steady gaze of the court of law[20]. The initial proclamation acquaints the debate with the jury individuals and goes about as a general guide on how the whole preliminary would be unfurled. An end proclamation then again contains the rundown of the procedures and helps the jury to remember the key proof that has been introduced so the jury can be convinced to give a good decision[21]. 9. The starting point of the Uniform Evidence Law can be followed back to the request which started in the year 1979 of the ALRC, i.e., the Australian Law Reform Commission, which was accused of the audit of the relevant laws of proof to the procedures of the courts. NSWLRC or the New South Wales Law Reform Commission had its own request in this issue. Their work was suspended in the year 1979 as the ALRCs audit result was pending. The last report of NSWLRC was created in 1988 which required that in New South Wales, the proposals of ALRC ought to be actualized. The Governments of New South Wales and the Commonwealth, in 1991, built up the bills to offer impact to the suggestions of the ALRC. Notwithstanding, none of these bills were passed. The Parliaments of New South Wales and the Commonwealth, in 1995, ordered new Evidence Acts. Both the ALRC and NSWLRC surveyed the activities of both the New South Wales and the Commonwealth Act. After an audit of the Evidence Act by both these, a report was distributed in February 2006. The Victorian Law Reform Commission likewise distributed a report after which, this law was formulated[22]. 10. The goal behind the presentation of the Evidence Act 2008 (Vic) was to present a demonstration which introduced a uniform proof law in Victoria. The avocation for drawing out this demonstration was to introduce new arrangements for law of proof, which were uniform with the law in such manner, with that of the New South Wales and the Commonwealth[23]. 11.The Evidence Act 2008 (Vic) has been partitioned into parts, inside which contains the pertinent arrangements to a specific part. For example, the utilization of the put on a good show 1.2, which contains all the arrangements identifying with this matter[24]. A case of this can be found in Section 7 of this demonstration, which ties the crown[25]. In this way, the significant arrangements on a specific subject have been clubbed together in parts. The Evidence Act 2008 (Vic) is identified with Evidence Act 1995 (Cth)[26], as it is a changed and better form of the 1995 demonstration. The arrangements of the later demonstration have a more extensive reach in contrast with the past enactment. Anyway in huge parts, the 2008 demonstration is uniform with the 1995 act[27]. Book index Cases R v Connors [2000] NSWCCA 470 at [133] R v Milton [2004] NSWCCA 195 Enactment Proof Act 1995 (Cth) Evidence Act 2008 (Vic) Others Dark A, Not-So-Uniform Evidence Law: Reforming Longman Warnings (2007) https://www.austlii.edu.au/au/diaries/CICrimJust/2007/25.pdf Legal College of Victoria, Introduction to the Uniform Evidence Act in Victoria: Significant Changes (2009) https://www.judicialcollege.vic.edu.au/locales/default/records/IntroductiontotheUEA.pdf Legal Commission of New South Wales, Checklist of Jury Directions (15 June 2015) https://www.judcom.nsw.gov.au/productions/benchbks/sexual_assault/checklist_of_jury_directions.html Rottenstein Law Group LLP, What are shutting explanations or shutting contentions? (2017) https://www.rotlaw.com/legitimate library/what-are-shutting explanations or-shutting contentions/ Rutkowski L, How to Prepare Yourself to Present Your Case (22 June 2015) https://www.peoples-law.org/how-plan yourself-present-your-case UN Women, Evidence assortment (2012) https://www.endvawnow.org/en/articles/1136-proof collection.html US Courts, Differences Betweeen Opening Statements Closing Arguments (2017) https://www.uscourts.gov/about-government courts/instructive assets/about-instructive effort/action assets/contrasts Victoria University, Bachelor of Laws: Evidence (2015) https://libraryguides.vu.edu.au/law/proof [Judicial Commission of New South Wales, Checklist of Jury Directions (15 June 2015) https://www.judcom.nsw.gov.au/productions/benchbks/sexual_assault/checklist_of_jury_directions.htmlAlice Gray, Not-So-Uniform Evidence Law: Reform