Structural Interconversion of Holin Transmembrane Domain I

Structural Interconversion of Holin Transmembrane Domain I Structural Interconversion of Holin Transmembrane Domain I is Dictated by a Single Proline: A FRET-based Analysis and its Functional Importance in Pore Formation. Muralikrishna Lella,†  Soumya Kamilla,†¡ Vikas Jain†¡,* and Radhakrishnan Mahalakshmi† ,* KEYWORDS (Word Style â€Å"BG_Keywords†). If you are submitting your paper to a journal that requires keywords, provide significant keywords to aid the reader in literature retrieval. ABSTRACT: Mycobacterial cell lysis during the lytic D29 bacteriophage infection is timed by perfect orchestration amongst/between components of the holin-endolysin cassette. In Gram-negative bacteria, progressively accumulating holin in the inner membrane, retained in its inactive form by anti-holin, is triggered into active hole formation, resulting in the canonical host cell lysis. However, the molecular mechanism of regulation and physical basis of pore formation in the mycobacterial inner membrane by D29 holin, particularly in the nonexistence of an anti-holin, is poorly understood. In this study, we report, for the first time, the use of fluorescence resonance transfer measurements to demonstrate that the first transmembrane domain (TM1) of D29 holin undergoes a helix ↔ ÃŽ ²-hairpin conformational interconversion. We validate that this structural malleability is mediated by a centrally positioned proline, and is abolished in the conformationally rigid substitution mutants containing Ala, DPro, or Aib. Using electrophysiology measurements coupled with calorimetric vesicle assays, we demonstrate that due to the conformational switch, native TM1 exhibits sluggish self-association in membrana, while its rigid variants show accelerated lipid bilayer disruption. The biological implications of D29 holin structural alteration is presented as a holin self-regulatory mechanism and its implications are discussed in the context of data-driven peptide-based therapeutics. The fatal host cell lysis step during bacteriophage infection is one of the most precisely programmed events, coordinated by the mechanical membrane disruption by a hole-forming membrane protein termed holin and the peptidoglycan-degrading enzyme endolysin.1 In the canonical holin-endolysin cassette, holin accumulates in the bacterial inner membrane and is retained in the inactive form until membrane depolarization drives holin assembly into holes that are large enough for endolysin release.1b,2 It is believed that ion leak through pinholes formed in holin-enriched lipid rafts would result in local membrane depolarization, which would exponentially propagate throughout the bacterial inner membrane and result in the formation of >300 nm diameter holes.2g,3 Historically, genetics of the coliphages T4, ÃŽ », and 21 have been extensively investigated, and therefore our current understanding of holin function and regulation is largely derived from the lysis effector S105 (or S2168) holin and the antiholin S107 (or S2171).2d,2g,4 Based on more recent functional characterization of members from eight holin superfamilies comprising several bacteriophages,5 we now know that the number of transmembrane ÃŽ ±-helical segments (TMSs) can vary from 1-4; of this, the 3-TMS is widely prevalent.1b,6 Despite conceptual and experimental advances in our understanding of holin function, very few studies have translated these findings to the mechanism of holin regulation in mycobacteriophages (Mφ). Currently, >4000 documented Mφ species exist, of which >600 have been sequenced.7 Of particular interest is the lytic Mφ D29, which is the predator for Mycobacterium tuberculosis, among other mycobacteria. Mφ D29 possesses a putative holin sequence coded by the gp11 gene, and is predicted to possess two transmembrane segments, typically observed in class II holins.8 However, an antiholin sequence is conspicuously missing in the Mφ D29 genome, raising concerns on how this phage achieves holin regulation. A previous finding from our laboratory demonstrated that the first transmembrane domain (TM1) of D29 Mφ holin could undergo a conformational switch from a helical form to an extended structure, and a centrally located Pro-Gly segment was important for such interconversion.9 This opened further questions on the biophysical nature of such a conformational conversion, the functional implications during holin assembly and whether such interconversion did indeed possess any regulational role within the mycobacterial cell. In this study, we demonstrate that the D29 Mφ TM1 undergoes a helix ÃŽ ²-hairpin conversion that is abolished in Pro -> Ala/DPro/Aib mutation. We also show that proline internally regulates assembly of TM1 in the membrane, and could potentially function as the ‘missing’ antiholin in D29 Mφ. RESULTS AND DISCUSSION ASD ASD METHODS Peptide synthesis and labeling with fluorescent probes. All peptides were synthesized using Fmoc chemistry on a Rink Amide AM resin with a 0.63 mmol/g loading capacity, using DMF as the medium. Deprotection of Fmoc was achieved using 20% piperidine and the progress of the reaction was monitored using Kaiser test and mass spectrometry.9-10 Final peptide was generated using the cleavage cocktail (TFA : water : phenol : ethanedithiol : thioanisole in the ratio 85:5:5:2.5:2.5), followed by cold ether precipitation, and verified by mass spectrometry. On-resin labeling of the fluorophore (Alexa Fluor ® 350 or dansyl chloride) at the N-terminal residue was achieved using HOBt or DIPEA in DMF. All labeling reactions were carried out at least twice and confirmed by mass spectrometry. Labeling efficiency was calculated using labeled peptide absorbance at fluorophore ÃŽ »max ­ and unlabeled : labeled peptide ratios for all reactions were maintained at ~1.0:0.5. Details are in the electronic supplementary information (ESI). Peptide folding and circular dichroism experiments. Desired quantity of peptide in the powder form was dissolved in 100 mM LDAO (lauryldimethylamine oxide) or 100 mM DPC (n-dodeyclphosphocholine) micelles prepared in 50 mM sodium phosphate pH 7.2, and were subjected to repeated cycles of heating and vortexing to promote peptide folding.9 All biophysical experiments were carried out using 0.022-0.024 mM samples, unless otherwise specified. Quantification was achieved using a molar extinction coefficient of 8408 M-1 cm-1 at 280 nm.CD spectra were acquired in various micellar conditions at 25  °C, using a 1 mm path length quartz cuvette at scan speeds of 100 nm/min. Data were integrated over three acquisitions and converted to molar ellipticity values using reported methods.9,11 Thermal denaturation and recovery measurements were carried out between 5-95  °C and 95-5  °C, respectively at a ramp rate of 1  °C/min. Details are provided in the ESI. Fluorescence and anisotropy measurements. Steady state Fà ¶rster Resonance Energy Transfer (FRET) measurements were carried out using Trp excitation at 280 nm ( ±2 nm slit width) and emission spectra were recorded between 295-550 nm ( ±3 nm slit width). Inter- and intra-molecular FRET was demarcated by titrating unlabeled peptide into labeled peptide samples to achieve stepwise dilutions and final unlabeled: labeled ratios of 1:1, 1:0.8, 1:0.6, 1:0.4, 1:0.2, 1:0. Data were normalized against Trp emission intensities and acceptor intensity at ÃŽ »max ­ were plotted (Alexa Fluor ® 350 ÃŽ »max = 442 nm and dansyl chloride ÃŽ »max = 500 nm). Anisotropy values were acquired using ÃŽ »ex-max = 442 nm and ÃŽ »em = 345 nm for both the labeled and unlabeled peptides. Lifetime measurements were carried out using time correlated single photon counting. Trp excitation was achieved at 292 nm using a pulsed LED and fluorescence decays were monitored at the ÃŽ »em-max for the respective samples (345 nm in LDAO; 347 nm in DPC; 355 nm in buffer). All data were fitted to a triple exponential decay to derive lifetimes (Ï„i) and their respective amplitudes (ÃŽ ±i). The average lifetime was given as = ÃŽ £Ãâ€ži.ÃŽ ±i.12Details are provided in the ESI. Pore formation measurement using planar lipid bilayers. Black lipid membranes were generated using DiPhPC (diphytanoyl phosphatidylcholine) on a planar lipid bilayer workstation in which the membrane bilayer was painted across a 150 ÃŽ ¼m aperture generated in the septum of a Delrin cup. A constant 10 mV voltage was applied in both cis and trans sides of the chamber, pre-filled with mM sodium phosphate pH 7.2 containing 0.5 M KCl. 0.022-0.024 mM peptide was added to the cis chamber and electric current was recorded using a 50 Hz filter, sampling frequency of 10 kHz, and digitized. Opening and closing event frequency was calculated throughout the recording and converted to conductance using the formula: [observed current in pA] / 10 mV = conductance in nS. Details are described in the ESI. ASSOCIATED CONTENT (Word Style â€Å"TE_Supporting_Information†). Supporting Information. A brief statement in nonsentence format listing the contents of material supplied as Supporting Information should be included, ending with â€Å"This material is available free of charge via the Internet at http://pubs.acs.org.† For instructions on what should be included in the Supporting Information as well as how to prepare this material for publication, refer to the journal’s Instructions for Authors. ABBREVIATIONS CCR2, CC chemokine receptor 2; CCL2, CC chemokine ligand 2; CCR5, CC chemokine receptor 5; TLC, thin layer chromatography. REFERENCES (Word Style TF_References_Section). References are placed at the end of the manuscript. Authors are responsible for the accuracy and completeness of all references. Examples of the recommended formats for the various reference types can be found at http://pubs.acs.org/page/4authors/index.html. Detailed information on reference style can be found in The ACS Style Guide, available from Oxford Press. (1) (a) Smith, D. L.; Young, R. J. Bacteriol. 1998, 180, 4199-4211; (b) Young, R. Current Opinion in Microbiology 2013, 16, 790-797; (c) Young, R. Microbiological Reviews 1992, 56, 430-481. (2) (a) Young, R.; Wang, I.; Roof, W. D. Trends Microbiol. 2000, 8, 120-128; (b) Ramanculov, E.; Young, R. Gene 2001, 265, 25-36; (c) Wang, I. N.; Deaton, J.; Young, R. J. Bacteriol. 2003, 185, 779-787; (d) Park, T.; Struck, D. K.; Deaton, J. F.; Young, R. Proc. Natl. Acad. Sci. U. S. A. 2006, 103, 19713-19718; (e) Savva, C. G.; Dewey, J. S.; Deaton, J.; White, R. L.; Struck, D. K.; Holzenburg, A.; Young, R. Mol. Microbiol. 2008, 69, 784-793; (f) Pang, T.; Savva, C. G.; Fleming, K. G.; Struck, D. K.; Young, R. Proc. Natl. Acad. Sci. U. S. A. 2009, 106, 18966-18971; (g) White, R.; Chiba, S.; Pang, T.; Dewey, J. S.; Savva, C. G.; Holzenburg, A.; Pogliano, K.; Young, R. Proc. Natl. Acad. Sci. U. S. A. 2011, 108, 798-803; (h) Young, R. Journal of Microbiology 2014, 52, 243-258. (3) (a) Dewey, J. S.; Savva, C. G.; White, R. L.; Vitha, S.; Holzenburg, A.; Young, R. Proc. Natl. Acad. Sci. U. S. A. 2010, 107, 2219-2223; (b) Pang, T.; Fleming, T. C.; Pogliano, K.; Young, R. Proc. Natl. Acad. Sci. U. S. A. 2013, 110, E2054-2063; (c) Savva, C. G.; Dewey, J. S.; Moussa, S. H.; To, K. H.; Holzenburg, A.; Young, R. Mol. Microbiol. 2014, 91, 57-65. (4) Blasi, U.; Nam, K.; Hartz, D.; Gold, L.; Young, R. EMBO Journal 1989, 8, 3501-3510. (5) Reddy, B. L.; Saier, M. H., Jr. Biochim. Biophys. Acta 2013, 1828, 2654-2671. (6) Wang, I. N.; Smith, D. L.; Young, R. Annu. Rev. Microbiol. 2000, 54, 799-825. (7) http://phagesdb.org/. (8) (a) Catalao, M. J.; Gil, F.; Moniz-Pereira, J.; Pimentel, M. J. Bacteriol. 2011, 193, 2793-2803; (b) Hatfull, G. F.; Jacobs-Sera, D.; Lawrence, J. G.; Pope, W. H.; Russell, D. A.; Ko, C. C.; Weber, R. J.; Patel, M. C.; Germane, K. L.; Edgar, R. H.; Hoyte, N. N.; Bowman, C. A.; Tantoco, A. T.; Paladin, E. C.; Myers, M. S.; Smith, A. L.; Grace, M. S.; Pham, T. T.; OBrien, M. B.; Vogelsberger, A. M.; Hryckowian, A. J.; Wynalek, J. L.; Donis-Keller, H.; Bogel, M. W.; Peebles, C. L.; Cresawn, S. G.; Hendrix, R. W. J. Mol. Biol. 2010, 397, 119-143; (c) Payne, K.; Sun, Q.; Sacchettini, J.; Hatfull, G. F. Mol. Microbiol. 2009, 73, 367-381. (9) Lella, M.; Mahalakshmi, R. Chem. Commun. 2013, 49, 9594-9596. (10) Makwana, K. M.; Raghothama, S.; Mahalakshmi, R. Phys Chem Chem Phys 2013, 15, 15321-15324. (11) Maurya, S. R.; Chaturvedi, D.; Mahalakshmi, R. Sci. Rep. 2013, 3, 1989. (12) Maurya, S. R.; Mahalakshmi, R. J. Biol. Chem. 2013, 288, 25584-25592.

Post war Greece Essay

Benito Mussolini had expansionists policies in his Fascist regime in Italy. By the mid of 1940, Mussolini had started admiring Adolf Hitler’s conquests and wanted to prove his counterpart, Hitler who was an Axis partner, that he too could lead Italy to success in war. Italy took control over Albania in 1939. Italians invaded Greece after the Greek dictator whose name was I. Metaxas refused to honor Italian ultimatum demanding the occupation of Greek territory. The Greek counter attacked and forced the Italians out and even took control of Albania formerly under Italy. In April 1941, Germany started attacking Greece and the Italian army also resumed their attack to Greece. As a result, the Greece army started retreating back from Albania to avoid a possible cut off by the rapidly advancing Germany troops. On April 20th, the Greek army based at Epirus surrendered to their enemies,the Germans and on the 23rd the same month, the same was repeated now to include the Italians and thus bringing the war between Greece and Italy to an end. The Greek victory over the Italian offensive on October 1940 was the first victory of the second world war and this boosted the morale in the occupied Europe Italian soldiers came across the Greek border in the 28th day of October 1940 but Greek defenders who were more determined drove these invaders back in to Albania (McNail, 167). As the death of Metaxas came in January 1941, he had already undergone a transformation from unpopular dictator who was not admirable by the people in to a national leader liked by the people, by his defiance of Mussolini and to the people his death was a great loss. Hitler was forced to reluctantly divert the troops from Germany to go and rescue Mussolini from being defeated and as a result ended up attacking Greece through Bulgaria and Yugoslavia. In response, the Greek sought for assistance from the British, and assistance was given readily although Greeks kept insisting stubbornly to defend Macedonia and Thrace from attacks by the Germany invaders while the Greece’s only hope was to strategically withdraw to a defensive line on a river south of Thessaloniki. Towards the end of May, the German troops had taken control over much of the land in Greece. Both the king and the government escaped to Crete and stayed here until the battle of Crete finally came to an end. From here, they transferred to Egypt where they established a government in exile while an establishment of a Nazi held puppet regime took place in Athens, Greece. Members of this regime were either conservatives or belonging to the nationalists but with fascist leanings. The three people who collaborated with the enemy were T. Georgis, K. Logothetopoulos and R. Ioannis. T. Georgis was the general who signed the armistice earlier with Wehrmacht, while Logothetopoulos had become naughty for having aimed to recruit juvenile volunteers to help in restrengthening the Germany army. On the other hand, R. Ioannis had a very notable achievement in that he saw to the creation of the security battalions for the collaborationists to protect those who supported the enemies. Greece greatly suffered very terrible privations during the second world war when the Germans took control of most of the Greece’s agricultural production and also prevented the fishing fleets of Greece from operating. Following the Axis force occupation on the Greek land, a great famine struck in the year 1941 and 1942 because of the blockade by the British naval and also due to the fact that Germany soldiers had seized the crops. Due to hunger and other associated effects of famine, several thousands of Greeks died. AS a result, several resistance movements came up in the hilly and mountain regions and soon the Germans and those who supported them remained in control of only the major highways and the connected towns. The largest of the up coming group was the National Popular Liberation Army (the ELAS) which was under the control of the communists and a civil war soon broke out between the National popular liberation army and the non- communists uprising groups such as the national republican Greek league (the EDES) in the areas which had been liberated from the German troops. The royalist government set up in exile in Cairo was only intermittently keeping in touch with the resistant movement and this government in Cairo failed to appreciate how the monarchy set up in Greece had lost its popularity. The resistance of the Greeks people against invasion by the Italians had a great influence to the course the second world war took. Adolf Hitler would say that if Italy had not attacked Greece and needed Germany assistance, the world war would have taken a quite different course. What would be anticipated was the cold war in Russia by weeks, Moscow and Reningrad conquered and Stalingrad would not have existed. The need to occupy Greece, suppress partisans and defend Greece from Allied actions led Germany and Italy to drop much of their differences during the second world war. The Greek resistance ultimately necessitated collaboration between Germany and Italy. Due to political considerations, British forces were send to Greece from Middle East, and this was considered to be a major strategic mistake because that was a critical stage to divert military forces from Middle East. The British forces could not stop Germany invasion in Greece. Political disorder and the 1946-49 war in Greece In October 1944, German forces withdrew from Greece and the Greece government in exile came back to Athens. Then the ELAS guerrilla soldiers took control over most of the land in Greece with its leaders having an ultimate aim to control the whole country despite Stallin’s agreement that Greece would remain within the region of influence by the British after the war. Demonstrators from the Communist Athens ended in violence in early December 1944, and then followed a door to door battle with the British forces and the monarchist armies. Before a month had ended, the communists had been defeated and an unstable government of coalition was established. Continued tensions within this government led to a civil war in the year 1946. Greece was backed by Britain and later by the United States in terms of military support and economic aid. Between 1947 and 1948, the communist forces had conquered much of the Greece mainland and were able to move freely within the land. With the extensive material support from America and the reorganization, the Greek navy steadily took control over much of the mainland. Yugoslavia had to close her borders from the insurgent military in 1949 after it broke relationship with the Soviet Union. In 1949, a final offensive was launched by Marshal P. Alexander that forced the insurgents who were left back to flee across the border to the north in to the territories of the Greece neighbors or else they surrender. The civil war left about one hundred people killed and also resulted to major damages to the economy. More than twenty five thousand Greeks and many Macedonian Slavs were evacuated to the eastern bloc countries either by force or voluntarily while over seven hundred thousand people became internally displaced people in Greece and many more emigrated to, among other countries, Australia. This settlement after the war greatly expanded the Greece territory which had already started earlier in 1832. In 1947, a treaty in Paris was signed and it required Italy to surrender the Dodecanese islands to the Greece. These islands had the majority of inhabitants speaking Greece and were part of the last areas to be incorporated to the Greek state, except Cyprus which remained under possession of Britain until its independence later in 1960. Greeks ethnicity became more homogeneous after the war when more than twenty five thousand Albanians were expelled from Epirus. The remaining minorities of no significance were the Muslims in West Thrace and few Slavic speaking in the north. Greeks continued to claim more of the southern Albania where significant Greek population lived. Economic miracle for Greece: 1955-79) The Greek economic miracle implies the factual and impressive rate of social and economic development that occurred in Greece from the early of 1950s to the middle of the 1970s. The average rate of economic growth recorded between the year 1950 to 1973 was about seven percent which was the worlds second best after that of Japan in those times. The 1950s growth rate was the highest sometimes going above ten percent, almost nearing those of a modern tiger economy. This continued up to 1960s. The Greek people did not view it as a miracle because since the period after the war until the middle of the 1970s, it was a time of deep political divisions that resulted to military dictatorship between 1957 to 1964 and the Greeks did not see any positive economic change for this period of twenty years. Further more, growth initially only widened the economic gap between the few rich and the majority poor and this could only intensify political divisions. Between 1941 and 1944, the Axis occupation and the fighting with resistance groups had unexpected effects on the infrastructure and also on the Greece’s economy. Given also that after the end of the first world war, Greece went in to civil war, its economy had drastically fallen by 1950. The Greece per capita income as a measure of its purchasing power fell, just like that of France, from 62 percent to about 40 percent in the year 1949 (Cranidlis, 97). Greece experienced a rapid recovery of its poor economic condition. This was as a result of a number of factors among them the stimulation from the Marshall plan, a fast devaluation of the Drachma, more foreign investments, significant industrial development especially the development of the chemical industry, development of the public services sector and development of tourism industry and a widespread construction activities coming as a result of enormous infrastructural project building and rebuilding in the Greek towns and cities. The construction activities are connected with the fast economic growth on the society and the development of its towns. This led to renewal of the urban through replacing the pleasant urban, consisting mainly of low rising houses and peoples homes, with a continuous set of concrete and block storey and skyscrapers in most key cities and towns. After 1950, economic growth consistently was better than that of many European countries in terms of annual growth. The only time there was economic stagnation was in the 1980s but it was counterbalanced when the Greek black economy evolved at the same time. The good economic performance made Greece advance its economy to enjoy a per capita income almost the same as that of other European Union partners like France and German (Krofas, 123). After end of the civil war, Greece sought to join hands with the western democracies by becoming a member of NATO in the year 1952. From this time to the late 1963, Greece was under conservative parties. In1964, the party called the Center Union led by George Papandreau was elected and remained in rule until July of 1965 before he was dismissed by king Constantine II, thereby resulting to a constitutional crisis. The fall of this government led to a series of coalition governments between the conservatives and rebel liberals. On April 1967, a group of colonels from the right wing seized power in a coup. They suppressed civil liberties , established special military courts and dissolved political parties. Thousands of political party opponents and those suspected to be communists were imprisoned or taken to exile to very remote Greek islands. United States was alleged to have supported junta and this caused the rise of anti- Americanism in the Greece during and immediately after the junta rule. However, the united states had already earned the animosity of communists in Greece long before. In 1974, a referendum resulted in abolishment of a monarchy and a new constitution was made and passed by the parliament on the 19th of June 1975. President T. Constantine was elected by parliament to head the republic. In the 1977 elections to the parliament, the New democracy party won for a second time with a majority seats. In 1980, Karamanlis who was the prime minister was elected to succeed T. Constantine as the president with George Rallis being elected as the prime minister to succeed Tsatsos. Greece joined the European Community, now called the European Union on 1st January 1981. It also elected the country’s first socialist government. In 1989, there were two rounds of parliamentary elections which both produced coalition governments that were weak and had limited mandates. Party leaders held back their support for those governments and elections were once again held in April. The new democracy party led by Constantine Mitsotakis won. In 1992, Samaras formed his own party after being fired from the position of a cabinet minister for Foreign affairs. He called the party the Political Spring. As a result of this division, the New democracy government collapsed and when new elections were held in September 1993, Papandreou returned to power. Papandreou resigned on January 17th of 1996 due to a protracted illness. He was then replaced as the country’s prime minster by the former minister of trade and industry by the name, Costas Simitis who consecutively won the elections in 1996 and also in 2000 before retiring in 2004. His successor was George Papandreou as PASOK leader. In March 2004, elections were held and New Democracy under the leadership of Costas Karamanlis who was a nephew to the former president saw the victory over the PASOK. Instead of waiting for normal elections that should have been held in March the year 2008, the government called for elections in September 2007 and the New Democracy once again won the majority vote in the parliament. Due to this repeated defeat, the PASOK underwent a party election in search of a new leader that saw Mr. Georgis Papandreau re-election as the party leader of the Socialist Party in Greece. Review of Economic Literature It is important to review the economic thoughts from both the Greek and the American writers and compare them in the perspective of the economic reconstruction of Greece. A good approach would be first reviewing the economic thought in the 1930s and 1940s and then that of the American technical experts who were either in Greece based American embassy or in the AMAG. (Lincoln, 64). Reviewing the plans by Batsis, Valvaresos and Zolotas is also of importance to acquire a clear thought about the economic miracle and general development for the Greece from very poor to one of the world’s best economies. A scrutiny of what the exact agenda of the AMAG and other aids to develop the Greece economy was, is of importance in shining light in to this issue. Establishing whether there was any difference between the American economic experts working with the AMAG and Americans working in the American embassy to Greece in Athens in terms of priorities and the paths to reconstruction is felt to be essential. This way, most questions of much concern to economic historians will be answered well and also valuable material towards making reconstructions in the history of economics will be unearthed, especially for the post second world war period in Greece The depression of economy in the whole world led the default in Greece in the year 1932and afterwards to the introduction of a quite successful system. The gross domestic product was 510 million dollars in the year 1931, then dropped to 330 million dollars in the year 1932, remained at the same stable low level in 1934 at 340 million dollars and drastically rose in the subsequent years of 1934 at 490 million dollars, 1935 at 510 million dollars and 1936 at 550 million dollars. Despite the economic success, the poor remained miserable due to the falling prices and the problem of the merchant class which was used to free trade and had problem with coping in the new trade conditions of exchange controls. Most Greek economists concentrated on writing and analyzing the economic success of their country and did very little on questioning the actual role of the American Mission Aid for Greece that it played in uplifting the economic situation of Greece. The economists made numerous economic publications such as books, journals, treatises, periodicals among others. The interventionists were most influential and they had studies in Germany in 1920s to 1930s. These economists espoused a development theory stage to which the Greek government ought to take to speed up the rate of economic development. It was based on the fact that the economy of Greece could not rely on private initiative alone. Most of them remained hostile to abstract thinking and viewed the dirigiste policies world over as evidence of the validity of their thesis. The liberals viewed the historical present hitherto as a parenthesis and had a believe that the whole world including Greece economy ought to have an international economic corporation. The liberals also supported that it was necessary for the state to intervene and as a result they came up with a frame upon which the Greece government should act upon in a liberal economy. The Marxists viewed the crash of 1929 as evidence for capitalism break down. Particularly in Greece, the imperialism of the great powers and the kings rule were to be overthrown by mass action for socialism to be established.

Accounting Analysis Essay - 2047 Words

1. Accounting Analysis Assess the degree to which the firm’s accounting reflects the underlying business reality. Identify accounting distortions and evaluate their impact on profits and the sustainability of profits. Financial statements are used to determine the business activities of a firm and the role of accounting analysis is to determine the accuracy and quality of the information provided. This analysis would look into the degree of its accounting figures captures its business reality through the policies used and its resulting noise, potential forecast errors and its impact on Myer’s profit. A few critical areas which are vital to Myers business which includes credit losses, quality of net accounts receivable, inventory†¦show more content†¦This increase in flexibility would allow Myer to increase its accuracy in terms of the term it is used for. This could possibly explain the increase in depreciation in 2011 of 79443k compared to that in 2010 of 62705k. Intangible assets are one of the most significant items in Myers financial statement. It consists of goodwill, brand names and trademarks, software and leases. AASB 136 Impairment of Assets requires Goodwill and some of the brand names that are indefinite useful life to test for the impairment. In Myer, there is no impairment loss. Furthermore, the accumulated amortisations of the other intangible assets are shown in the table X have a total value of $73585 thousand. According to AASB 117 Leases, the total rentals leases over the leases term are being expensed on a straight-line basis. In contrast, Myer’s competitor David Jones has only two intangible assets goodwill and software. The accumulated amortisation for software is $28808 thousand which is shown in the table X and it is the total value of accumulated amortisation. David Jones does not included leases as part of their intangible assets but Myer does. 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PG Japan the Sk-Ii Globalization Project - 1692 Words

1. As Paolo DeCesare, what factors do you need to consider before deciding what to recommend in your SK-II presentation to the global leadership team (GLT)? What kind of analysis will you need to do in preparing for that meeting? There are many factors that need to be considered when deciding recommendations to the GLT of SK-II. Being that this is PG’s first proposal to build a Japanese brand worldwide, it is important to take into consideration different cultures than of the consumers in Japan. Since 2005, PGs global organization has been in the middle of a restructuring program, which can sometimes cause a disruption to the normal functions of the company. Local managers resisted the surfacing of new products based on the†¦show more content†¦On the other hand, being that PG is a somewhat new company in China, their knowledge may be limited in the Chinese culture. And, all of SK-II’s other competitors are already offered in that market. So, the question arises – what is SK-II’s market readiness? To the customer, they are not familiar with this product’s application. They usually only apply a one-step program and have only recently started to switch to a three-step process. So, the seven-step application of a product might appear tedious to a customer. China also has the reputation for selling counterfeit products. The market is flooded with copies of upscale products, which causes increased costs for import taxes. DeCaesare’s second option highlights that the target market for the product consists of a more affluent clientele of beauty conscious people; and Europe has a large number of customers that fit in this category. In the Western market, SK-II’s target audience is already using a six to eight-step skin care process. Unfortunately for SK-II, this market is overcrowded and PG would need to find a way to create market visibility. Brand awareness in Europe historically shows to be costly due to television and print ad prices. Option three, for restructuring the brand in Japan, PG knows Japanese women are the most familiar, knowledgeable and are the leading consumers for the SK-II skin care solutions. It shows that Japan has the highest number of beauty-conscious consumers, which proves thatShow MoreRelatedPG Japan the Sk-Ii Globalization Project2059 Words   |  9 PagesGMBA 5075 International Management Case 5-2 - Pamp;G Japan: The SK-II Globalization Project Group Report Due date: 12/03/12 By: Wendy Rodriguez Francisco Oliveira Francisco Andujar Yichen Li Intro: This case describes how SK-II which is a fast-growing skin care product is becoming very popular with a price to match its performance. After being introduced in Hong Kong and in Taiwan, Pamp;G believes that this brand has a strong global potential. At the conclusion of this case, theRead MorePG Japan the Sk-Ii Globalization Project3489 Words   |  14 PagesPG Japan: The SK-II Globalization Project When looking at the archetype of PG it can be seen that it is an Multi Centred MNE. Which consists of a set of entrepreneurial subsidiaries abroad which are key to knowledge-based FSA development. National responsiveness is the foundation of the international strategy. The non-location bound FSAs that hold these firms together are minimal: common financial governance and the identity and specific business interest of the founders or main owners. LaterRead MorePg Japan1454 Words   |  6 Pagesto global business management. In the context of these changes introduced by Durk Jager, Pamp;G’s new CEO, Paolo de Cesare is transferred to Japan, where he takes over the recently turned-around beauty care business. Within the familiar Max Factor portfolio he inherits is SK-II, a fast-growing, highly profitable skin care product developed in Japan. Priced at over $100 a bottle, this is not a typical Pamp;G product, but its successful introduction in Taiwan and Hong Kong has de Cesare thinking theRead MoreSK-II, Japan1844 Words   |  8 Pagesï » ¿PG Japan: The SK-II Globalization Project (Harvard Business School case) Case Summary In â€Å"PG Japan: The SK-II Globalization Project† case study, the author Christopher Bartlett presents the PG’s plan of pushing SK-II as a global beauty product. In late 1999, Paolo de Cesare, President of Max Factor Japan, had given an idea to the Global Leader Team (GLT) of PG’s Beauty Care Global Business Unit (GBU) that whether it was a good idea for pushing SK-II to become a global PG brand. Since theRead MorePG Japan1409 Words   |  6 Pages Does SK-II have the potential to become a global brand within Procter Gamble’s worldwide operations? Why or why not? 2. Which of the three market options should Paulo Decesare recommendation to the GLT? What benefits do you expect to gain? What risks do you see? 3. How Should he implement your recommended option? What are the implications for PG’s new post-O2005 organization? What support and/ or resistance do you expect? How will you manage it? 4. Why was SK-II so successfulRead MorePG Japan1395 Words   |  6 Pages Does SK-II have the potential to become a global brand within Procter Gamble’s worldwide operations? Why or why not? 2. Which of the three market options should Paulo Decesare recommendation to the GLT? What benefits do you expect to gain? What risks do you see? 3. How Should he implement your recommended option? What are the implications for PG’s new post-O2005 organization? What support and/ or resistance do you expect? How will you manage it? 4. Why was SK-II so successfulRead MorePG Japan; Sk-21762 Words   |  8 Pages* Case Paper: Pamp;G Japan: The SK-II Globalization Project Mina Uwakubo Hawaii Pacific University Mr. Louis R. Collazo MGMT 6310 April 8, 2012 SK-II’s success is not only prestige skin care product or advanced technology but also its marketing approach to build the new brand. Pamp;G succeeded to connect between the core technology or product concept and local market. Through Japanese market among the world’s toughest competitors, Pamp;G developed potential source of innovations. InRead MorePG Case Study2393 Words   |  10 PagesSummary Statement/Abstract Pamp;G is a global company that is involved in many different markets including beauty and grooming and household care. The company has been through a lot of change due to the O2005 project that was undertaken in 1999 and saw a complete overhaul of the company during those years. One product that also was involved a lot during that time was the SK-II skin care product which was popular in Japan and a man named de Cesare wanted to take the product global to either theRead MoreGege997 Words   |  4 PagesAcquiring a Global Footprint (written by Tarun Khanna, Krishna G. Palepu, and Richard J. Bullock) 1. Should Tata Motors bid for Ford’s Land Rover and Jaguar units as part of its globalization efforts? What are pros and cons? 2. How else could Tata Motors compete with ford, Honda, Toyota, and other rivals? Could Tata Motors focus on competing as a parts supplier instead? 3. What is your assessment of the globalization and Mamp;A strategies of the Tata Group operating companiesRead MorePG Corparate10304 Words   |  42 PagesStatement (Actual) 4 Mission Statement (Proposed) 4 Vision Statement (Proposed) 5 Slogan (Proposed) 5 The CPM Matrix of Pamp;G Company 6 External Factor Evaluation (EFE) Matrix for Procter and Gamble (Pamp;G) 9 The I/E matrix for Procter and Gamble (Pamp;G) 14 Internal Factor Evaluation (IFE) Matrix for Procter and Gamble (Pamp;G) 16 A SWOT Matrix of Pamp;G Company 21 Strengths 23 Weaknesses 23 Opportunities 24 Threats 24 The FOUR strategies 25 The SPACE Matrix 26 Calculation