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Philippine historiography has long ignored the significant but complex role played by the people at the margins of society during the Japanese Occupation, except for some rural movements such as the Hukbalahap in central Luzon. During the Second World War, besides the coercion and violence perpetrated or orchestrated by the Japanese occupying forces from 1942, the people of Leyte experienced many kinds and levels of violence, including among local factions. At the onset of the invasion and from late 1944, Leyte was also the site of major naval and land battles between the returning American forces and the Japanese army, each side seeking to incorporate locals in their campaigns. This essay traces violent episodes involving and among members of the local elite and masses alike in Leyte, during and in the aftermath of the Japanese Occupation and the return of Americans up to Philippine Independence, to show how such violence was not only unleashed by war, but also had deep and complicated roots in colonial history, local politics and rural poverty.

The oleaginous yeast Lipomyces starkeyi has considerable potential in industrial application, since it can accumulate a large amount of triacylglycerol (TAG), which is produced from sugars under nitrogen limitation condition. However, the regulation of lipogenesis in L. starkeyi has not been investigated in depth. In this study, we compared the genome sequences of wild-type and mutants with increased TAG productivity, and identified a regulatory protein, LsSpt23p, which contributes to the regulation of TAG synthesis in L. starkeyi. L. starkeyi mutants overexpressing LsSPT23 had increased TAG productivity compared with the wild-type strain. Quantitative real-time PCR analysis showed that LsSpt23p upregulated the expression of GPD1, which encodes glycerol 3-phosphate dehydrogenase; the Kennedy pathway genes SCT1, SLC1, PAH1, DGA1, and DGA2; the citrate-mediated acyl-CoA synthesis pathway-related genes ACL1, ACL2, ACC1, FAS1, and FAS2; and OLE1, which encodes ∆9 fatty acid desaturase. Chromatin immunoprecipitation-quantitative PCR assays indicated that LsSpt23p acts as a direct regulator of SLC1 and PAH1, all the citrate-mediated acyl-CoA synthesis pathway–related genes, and OLE1. These results indicate that LsSpt23p regulates TAG synthesis. Phosphatidic acid is a common substrate of phosphatidic acid phosphohydrolase, which is used for TAG synthesis, and phosphatidate cytidylyltransferase 1 for phospholipid synthesis in the Kennedy pathway. LsSpt23p directly regulated PAH1 but did not affect the expression of CDS1, suggesting that the preferred route of carbon is the Pah1p-mediated TAG synthesis pathway under nitrogen limitation condition. The present study contributes to understanding the regulation of TAG synthesis, and will be valuable in future improvement of TAG productivity in oleaginous yeasts.Key pointsLsSpt23p was identified as a positive regulator of TAG biosynthesisLsSPT23 overexpression enhanced TAG biosynthesis gene expression and TAG productionLsSPT23M1108Toverexpression mutant showed fivefold higher TAG production than control

Fatty acid desaturases play vital roles in the synthesis of unsaturated fatty acids. In this study, Δ12 and Δ12/Δ15 fatty acid desaturases of the oleaginous yeast Lipomyces starkeyi, termed LsFad2 and LsFad3, respectively, were identified and characterized. Saccharomyces cerevisiae expressing LsFAD2 converted oleic acid (C18:1) to linoleic acid (C18:2), while a strain of LsFAD3-expressing S. cerevisiae converted oleic acid to linoleic acid, and linoleic acid to α-linolenic acid (C18:3), indicating that LsFad2 and LsFad3 were Δ12 and bifunctional Δ12/Δ15 fatty acid desaturases, respectively. The overexpression of LsFAD2 in L. starkeyi caused an accumulation of linoleic acid and a reduction in oleic acid levels. In contrast, overexpression of LsFAD3 induced the production of α-linolenic acid. Deletion of LsFAD2 and LsFAD3 induced the accumulation of oleic acid and linoleic acid, respectively. Our findings are significant for the commercial production of polyunsaturated fatty acids, such as ω-3 polyunsaturated fatty acids, in L. starkeyi.

Microbial lipids are sustainable feedstock for the production of oleochemicals and biodiesel. Oleaginous yeasts have recently been proposed as alternative lipid producers to plants and animals to promote sustainability in the chemical and fuel industries. The oleaginous yeast Lipomyces starkeyi has great industrial potential as an excellent lipid producer. However, improvement of its lipid productivity is essential for the cost-effective production of oleochemicals and fuels. Genetic and metabolic engineering of L. starkeyi via gene manipulation techniques may result in improvements in lipid production and our understanding of the mechanisms behind lipid biosynthesis pathways. We previously described an integrative transformation system using a drug-resistant marker for L. starkeyi . However, gene-targeting frequencies were very low because non-homologous recombination is probably predominant in L. starkeyi . Genetic engineering tools for L. starkeyi have not been sufficiently developed. In this study, we describe a new genetic tool and its application in L. starkeyi . To develop a highly efficient gene-targeting system for L. starkeyi , we constructed a series of mutants by disrupting genes for LsKu70p, LsKu80p, and/or LsLig4p, which share homology with other yeasts Ku70p, Ku80p, and Lig4p, respectively, being involved in non-homologous end-joining pathway. Deletion of the LsLIG4 gene dramatically improved the homologous recombination efficiency (80.0%) at the LsURA3 locus compared with that in the wild-type strain (1.4%), when 2000-bp homologous flanking regions were used. The homologous recombination efficiencies of the double mutant ∆ l sku70 ∆ lslig4 and the triple mutant ∆ lsku70 ∆ lsku80 ∆ lslig4 were also markedly enhanced. Therefore, the L. starkeyi ∆ lslig4 background strains have promise as efficient recipient strains for genetic and metabolic engineering approaches in this yeast.

Some historians are skeptical of Emilio Aguinaldo's heroism after his role in the Philippine revolution against Spain and the Philippine–American War in large part because of his apparent collaboration with the Americans and the Japanese. Actually little is known about events in Aguinaldo's life after 1899. This article aims to shed light on the obscured periods in Aguinaldo's life and career. In doing so, it addresses the questions: How could he have desired Philippine independence despite his submission to two imperial powers, America and Japan? Why did he collaborate with both powers? How would his place in Philippine history be evaluated?

The oleaginous yeast Lipomyces starkeyi is an excellent producer of triacylglycerol (TAG) as a feedstock for biodiesel production. To understand the regulation of TAG synthesis, we attempted to isolate mutants with decreased lipid productivity and analyze the expression of TAG synthesis-related genes in this study. A mutant with greatly decreased lipid productivity, sr22, was obtained by an effective screening method using Percoll density gradient centrifugation. The expression of citrate-mediated acyl-CoA synthesis-related genes (ACL1, ACL2, ACC1, FAS1, and FAS2) was decreased in the sr22 mutant compared with that of the wild-type strain. Together with a notion that L. starkeyi mutants with increased lipid productivities had increased gene expression, there was a correlation between the expression of these genes and TAG synthesis. To clarify the importance of citrate-mediated acyl-CoA synthesis pathway on TAG synthesis, we also constructed a strain with no ATP-citrate lyase responsible for the first reaction of citrate-mediated acyl-CoA synthesis and investigated the importance of ATP-citrate lyase on TAG synthesis. The ATP-citrate lyase was required for the promotion of cell growth and TAG synthesis in a glucose medium. This study may provide opportunities for the development of an efficient TAG synthesis for biodiesel production.

This article presents Local history in Leyte, Philippines, during the Japanese occupation, examining in particular the nature of collaborationism by Ormoc Mayor Catalino Hermosilla. In the context of local political feuds, this study shows how Hermosilla's collaboration with the Japanese enabled him not so much to advance Japanese goals but to strengthen his political power base. However, contrary to the view that the Japanese occupation did not alter social dynamics, this study shows that, although Ormoc's oligarchy remained conspicuous after the war, an important change had occurred as seen in the waning of Hermosilla's clout in municipal politics.

This article analyses the problem of food supply in Leyte, Philippines, during the Japanese occupation, which has not been studied in depth so far. It focuses on the interaction that took place among the Japanese occupying forces, anti-Japanese guerrilla groups, the Filipino collaborators, and the local residents over the procurement of foodstuffs. It also aims at clarifying the factors contributing to the disruption of the policy formulated by the Japanese and the Filipinos on the island. It is apparent in this study that the political and social characteristics in the province as well as the agricultural depression inherited from the American colonisation period brought about an outcome, which was different from the policy implemented in Manila.

The oleaginous yeast Lipomyces starkeyi is an attractive organism for the industrial production of lipids; however, the amount of lipid produced by wild-type L. starkeyi is insufficient. The study aims to obtain L. starkeyi mutants that rapidly accumulate large amounts of triacylglycerol (TAG). Mutagenized yeast cells at the early stages of cultivation were subjected to Percoll density gradient centrifugation; cells with increased production of TAG were expected to be enriched in the resultant upper fraction because of their lower density. Among 120 candidates from the upper fractions, five mutants were isolated that accumulated higher amounts of TAG. Moreover, when omitting cells with mucoid colony morphology, 11 objective mutants from 11 candidates from the upper fraction were effectively (100%) isolated. Of total 16 mutants obtained, detailed characterization of five mutants was performed to reveal that five mutants achieved about 1.5–2.0 times TAG concentration (4.7–6.0 g/L) as compared with the wild-type strain (3.6 g/L) at day 5. Among these five mutants, strain E15 was the best for industrial use because only strain E15 showed significantly higher TAG concentration as well as significantly higher degree of lipid to glucose and biomass to glucose yields than the wild-type strain. Thus, Percoll density gradient centrifugation is an effective method to isolate mutant cells that rapidly accumulate large amounts of TAG. It is expected that by repeating this procedure as part of a yeast-breeding program, L. starkeyi mutants suitable for industrial lipid production can be easily and effectively obtained.