Towards regulation of similar biotherapeutic products: Thailand's perspective

The implementation of universal health coverage scheme in Thailand allows quality, equitable and accessible health care for all. Patients with life threatening and chronic diseases can get access to biotherapeutic products to treat their ailments. This triggered a major impact on the need for specific guidelines in evaluation of similar biotherapeutic products in order to standardize the regulatory pathway to license this class of products ensuring that the products meet acceptable levels of quality, safety and efficacy. The development of similar biotherapeutic products (SBP) should be considered to ensure therapeutic equivalence of biotherapeutics products at more affordable prices. This will lead to greater ease and speed of approval and assurance of the quality, safety and efficacy of these products. Therefore, we report herein the SBP situation in Thailand.     

Structural analysis of a peptide synthetase gene required for ergopeptine production in the endophytic fungus Neotyphodium lolii.

Lysergyl peptide synthetase 1 catalyzes the assembly of toxic ergopeptines from activated D-lysergic acid and three amino acids. The gene encoding this enzyme in the endophytic fungus Neotyphodium lolii was analyzed and compared to a homologous gene from the ergot fungus Claviceps purpurea. Each gene contained two introns, which were found in the same relative position within two modules of the gene. The 5' ends of the two genes were unusually divergent. Signature sequences determining substrate specificity were similar in adenylation domains that recognized identical amino acids but differed within the adenylation domain for the amino acid that varies between the major ergopeptines of the two fungi. Homologues were detected in several related endophytic fungi; the tall fescue endophyte Neotyphodium coenophialum contained a divergent, second copy of the gene. Our results provide new information on the structure and distribution of this important peptide synthetase involved in ergot alkaloid biosynthesis.     

Touchdown-touchup nested PCR for low-copy gene detection of benzimidazole-susceptible Wuchereria bancrofti with a Wolbachia endosymbiont imported by migrant carriers

A novel, sensitive and specific touchdown–touchup nested PCR (TNPCR) technique based on two useful molecular markers, a Wuchereria bancrofti β-tubulin gene involved in benzimidazole susceptibility and a Wolbachia ftsZ gene involved in cell division, was developed to simultaneously detect the parasite W. bancrofti (W1) with its Wolbachia endosymbiont (W2) from both microfilaremic and post-treatment samples of at-risk migrant carriers infected with geographical W. bancrofti isolates. The detection and characterization of authentically low-copy gene-derived amplicons revealed no false positive identifications in amicrofilaremia with or without antigenemia. The W1-TNPCR was 100-fold more sensitive than the W2-TNPCR regardless of the microfilarial DNA isolation method and compared well with the thick blood film and membrane filtration techniques. These locus-specific TNPCRs could also detect Wolbachia-carrying W. bancrofti genotype in addition to a link to benzimidazole sensitivity among those with unknown infection origins that exhibited microfilaremia responsiveness against treatment with diethylcarbamazine plus albendazole. These TNPCR methods can augment the results of microscopic detection of the parasite because these methods enhance DNA isolation and PCR amplification capabilities.     

Multi-gene phylogenetic analyses reveals <Emphasis Type="Italic">Neohelicosporium</Emphasis> gen. nov. and five new species of helicosporous hyphomycetes from aquatic habitats

Helicosporous hyphomycetes are a morphologically allied group of Tubeufiales. We introduce a new helicosporous genus, Neohelicosporium, with five new species, Neohelicosporium aquaticum, N. guangxiense, N. hyalosporum, N. parvisporum, and N. thailandicum, based on morphological and phylogenetic evidence. The RPB2 protein gene data are provided to analyze their phylogeny in Tubeufiales. Phylogenetic analyses of combined ITS, LSU, RPB2, and TEF1α sequence data from 13 new isolates of Neohelicosporium provided evidence to support the establishment of the new taxa. The morphological characters of Neohelicosporium that differentiate it from other helicosporous species are compared and discussed.     

直翅目虫草（肉座菌目）：物种多样性及其与寄主的关系

迄今为止,以直翅目昆虫为寄主的虫草共报道25种,本研究详细介绍了它们的种类、寄主与分布,并给出了相关的分类学注释。ITS和tef1-α基因联合数据的分子系统发育显示,直翅目虫草分别隶属于白僵菌属Beauveria、绿僵菌属Metarhizium和线虫草属Ophiocordyceps 3个属。基于直翅目昆虫六大类群的生态习性特点,分析并讨论了直翅目虫草物种多样性与寄主的关系。以两种白僵菌Beauveria及戴氏虫草Metacordyceps taii（贵州绿僵菌Metarhizium guizhouense的异名）为例,介绍了虫草的根状菌索,一种介于子实体和孢梗束形体之间的根须状结构。种类修订：蟋螽白僵菌新组合Beauveria grylli comb. nov. ≡ 蟋蟀虫草Cordyceps grylli = Beauveria loeiensis,Beauveria kirkii comb. nov. ≡ Cordyceps kirkii。寄主更正：蟋螽白僵菌的寄主由原来的蟋蟀成虫（蟋蟀科Gryllidae）更正为螽斯（蟋螽）若虫（蟋螽科Gryllacrididae）,蟋蟀线虫草Ophiocordyceps tettigonia 寄主由原来的螽斯成虫（螽斯科Tettigoniidae：螽斯属Tettigonia）更正为蟋蟀成虫（蟋蟀科）。     

Direct ethanol production from cassava pulp using a surface-engineered yeast strain co-displaying two amylases, two cellulases, and β-glucosidase

In order to develop a method for producing fuel ethanol from cassava pulp using cell surface engineering (arming) technology, an arming yeast co-displaying α-amylase (α-AM), glucoamylase, endoglucanase, cellobiohydrase, and β-glucosidase on the surface of the yeast cells was constructed. The novel yeast strain, possessing the activities of all enzymes, was able to produce ethanol directly from soluble starch, barley β-glucan, and acid-treated Avicel. Cassava is a major crop in Southeast Asia and used mainly for starch production. In the starch manufacturing process, large amounts of solid wastes, called cassava pulp, are produced. The major components of cassava pulp are starch (approximately 60%) and cellulose fiber (approximately 30%). We attempted simultaneous saccharification and ethanol fermentation of cassava pulp with this arming yeast. During fermentation, ethanol concentration increased as the starch and cellulose fiber substrates contained in the cassava pulp decreased. The results clearly showed that the arming yeast was able to produce ethanol directly from cassava pulp without addition of any hydrolytic enzymes.