Fish oil improves motor function,limits blood–brain barrier disruption,and reduces Mmp9 gene expression in a rat model of juvenile traumatic brain injury

The effects of an oral fish oil treatment regimen on sensorimotor, blood–brain barrier, and biochemical outcomes of traumatic brain injury (TBI) were investigated in a juvenile rat model. Seventeen-day old Long-Evans rats were given a 15 mL/kg fish oil (2.01 g/kg EPA, 1.34 g/kg DHA) or soybean oil dose via oral gavage 30 min prior to being subjected to a controlled cortical impact injury or sham surgery, followed by daily doses for seven days. Fish oil treatment resulted in less severe hindlimb deficits after TBI as assessed with the beam walk test, decreased cerebral IgG infiltration, and decreased TBI-induced expression of the Mmp9 gene one day after injury. These results indicate that fish oil improved functional outcome after TBI resulting, at least in part from decreased disruption of the blood–brain barrier through a mechanism that includes attenuation of TBI-induced expression of Mmp9.     

An insect farnesyl phosphatase homologous to the N-terminal domain of soluble epoxide hydrolase

In insects, farnesyl pyrophosphate (FPP) is converted to juvenile hormone (JH) via a conserved pathway consisting of isoprenoid-derived metabolites. The first step of this pathway is presumed to be hydrolysis of FPP to farnesol in the ring gland. Based on alignment of putative phosphatases from Drosophila melanogaster with the phosphatase domain of soluble epoxide hydrolase, Phos2680 and Phos15739 with conserved phosphatase motifs were identified, cloned and purified. Both D. melanogaster phosphatases hydrolyzed para-nitrophenyl phosphate, however, Phos15739 also hydrolyzed FPP with a K_cat/K_m of 2.1 × 10⁵ M⁻¹ s⁻¹. RT-PCR analysis revealed that Phos15739 was expressed in the ring gland and its expression was correlated with JHIII titer during development of D. melanogaster. N-acetyl-S-geranylgeranyl-l-cysteine was found to be a potent inhibitor of Phos15739 with an IC₅₀ value of 4.4 μM. Thus, our data identify Phos15739 as a FPP phosphatase that likely catalyzes the hydrolysis of FPP to farnesol in D. melanogaster.     

Developmental morphology of the cyprinid fish, Candidia barbatus

 Embryonic, larval, and juvenile development of a Taiwanese cyprinid fish, Candidia barbatus, is described from laboratory-reared specimens. The eggs, measuring 1.8–2.1 mm in diameter, were demersal, almost spherical in shape, transparent and unpigmented, with a pale yellow yolk and no oil globule. Hatching occurred 56–69 h after fertilization, the newly hatched larvae measuring 4.9–5.3 mm in body length (BL) with 25–26 + 13–14 = 39–40 myomeres. The yolk was completely absorbed at 7.6 mm BL. Notochord flexion was initiated at 6.8 mm BL and finished at 7.6 mm BL. Aggregate numbers of all fin rays were completed at 12 mm BL. Barbels on the upper jaw appeared near the corner of the mouth at 17 mm BL. Eggs of the species closely resembled those of its related cyprinid genera, Opsariichthys and Zacco. Larvae and juveniles of C. barbatus were similar to those of O. uncirostris subspp., Z. platypus, and Z. pachycephalus, but differed from the latter in the process of disappearance of the adipose finfold (postflexion larval stage), barbels on upper jaw (juvenile stage), and pigmentation on the lateral body surface (postflexion larval and juvenile stages). Although C. barbatus also differed from the Z. temminckii species' group [Z. temminckii and Zacco sp. (sensu Hosoya, 2002)] in having barbels, larvae and juveniles of the former showed more similarity to the latter species group than to O. uncirostris subspp., Z. platypus, and Z. pachycephalus, from the aspect of head and body pigmentation.     

Early life history of a seahorse, Hippocampus mohnikei, in Tokyo Bay, Japan

The early morphological development, seasonal and spatial occurrence patterns, and food habits of a seahorse, Hippocampus mohnikei, in offshore waters of Tokyo Bay, central Japan, were studied on the basis of 206 juvenile and young specimens (6.0–65.3 mm TL) collected between August 1995 and January 1999. All the specimens were collected within the period from May to January, inclusive, each year. In the least developed specimen (6.0 mm TL), the number of dorsal, anal, and pectoral fin rays had attained the adult complement, whereas the minute caudal fin, consisting of two rays, was present in juveniles of 6.0–26.4 mm TL. Hippocampus mohnikei≧35 mm TL, being larger than settlement size (ca. 30 mm TL), had very low gut fullness index values (GFI = 0, >70% of specimens), whereas those of 15–29 mm TL had higher values (GFI = 2–4, >80% of specimens). In addition, larger individuals selectively fed on larger planktonic animals (species of Brachyura), which occurred naturally at low densities, although smaller food items, such as Oithona davisae and Penilia avirostris, occurred abundantly, being consumed by smaller H. mohnikei individuals (15–34 mm TL). These results indicated that food availability for H. mohnikei in offshore waters of Tokyo Bay is significant for individuals larger than settlement size, because their food preference would shift from smaller food items to larger food items, which would be scarce in their environments. Received: January 12, 2001 / Revised: May 13, 2001 / Accepted: June 14, 2001     

Growth of larval and juvenile Diaphus theta (Pisces: Myctophidae) in the transitional waters of the western North Pacific

Diaphus theta is one of the most common myctophid fish species in the subarctic and transitional waters of the North Pacific. The growth of larval and juvenile D. theta was investigated using sagittal otolith increment analysis of specimens caught in transitional waters of the western North Pacific. Samples taken over a 24-h period demonstrated that otoliths exhibited daily growth cycles, allowing accurate determination of age. Calcification of the incremental zone of otoliths took place only at night, suggesting that the formation cycle of the increment of juvenile D. theta was different from that of shallow-water fishes and would be related to their diel vertical migration. The relationships between standard length (SL) and daily growth increment (D) were expressed as linear equations: SL = 2.65 + 0.141D (r ² = 0.942) for larvae of 5.1–9.6 mm SL and SL = 3.54 + 0.129D (r ² = 0.933) for juveniles of 13.7–27.6 mm SL. The growth rates were 0.14 mm d⁻¹ in larvae and 0.13 mm d⁻¹ in juveniles; this is slow compared with tropical or subtropical mycto-phid species, in which growth occurs at about twice these rates. The larval period, including the metamorphic stage, was long compared with species at lower latitudes and was estimated to be 71 days. The slow growth rate and long period of larval stage of D. theta would be the life history pattern of high-latitudinal species adapted to a low-temperature habitat. Received: March 23, 2001 / Revised: July 5, 2001 / Accepted: July 19, 2001     

昆虫保幼激素促进家蚕杆状病毒系统的基因表达

杆状病毒表达载体系统(Baculovirus Expression VecterSvstem,BEVS)的一个最大优点是外源基因的高效表达(Hy-perexpression).但是,不同的外源基因在BEVS系统中的表达水平相差很大,较低的如α-干扰素,表达量为1～5mg/L培养细胞;高的如β-半乳糖苷酶,表达量可达600mg/L培养细胞.外源基因在BEVS系统中表达量受到诸多因素的影响,如细胞的类型与质量,外源基因蛋白的性质,启动子序列的完整性,是否为融合蛋白等[1].如何使外源基因在BEVS系统中高效表达,是近年来该领域中研究最活跃的方向之一.已证实家蚕杆状病毒的表达量受宿主遗传型的影响,最低和最高的遗传型相差达7倍以上[2].林水中等发现家蚕饲料中添食适当浓度的硫酸铜可提高外源基因单位表达量10%左右[3].杆状病毒在复制循环中表现出两种类型:芽生病毒和包涵体病毒,其中芽生病毒引起宿主体内不同组织间的感染,包涵体病毒则引起宿主之间感染[1].杆状病毒基因组中蜕皮激素尿苷二磷酸葡萄糖基转移酶(egt)基因影响激素在宿主体内的平衡[4],egt基因通过糖基化作用使蜕皮激素失活,打破宿主体内的激素平衡,延长幼虫期,以利于病毒的增殖[5].家蚕血淋巴中保幼激素(Juvenile hormone,JH)的滴度同样决定着幼虫发育的进程[6],本文通过体表使用保幼激素,以研究保幼激素对家蚕核型多角体病毒和宿主之间的相互关系及对外源基因表达量的影响.     

Short neuropeptide F (sNPF) is a stage-specific suppressor for juvenile hormone biosynthesis by corpora allata,and a critical factor for the initiation of insect metamorphosis

Molting and metamorphosis are essential events for arthropod development, and juvenile hormone (JH) and its precursors play critical roles for these events. We examined the regulation of JH biosynthesis by the corpora allata (CA) in Bombyx mori, and found that intact brain-corpora cardiaca (CC)–CA complexes produced a smaller amount of JH than that in CC–CA complexes and CA alone throughout the 4th and 5th (last) instar stadium. The smaller amount of synthesis was due to allatostatin-C (AST-C) produced by the brain. The CC synthesized short neuropeptide F (sNPF) that also suppressed the JH synthesis, but only in day 3 4th stadium and after the last larval ecdysis. For the suppression, both peptides prevented the expression of some of the distinct JH biosynthetic enzymes in the mevalonate pathway. Allatotropin (AT) stimulated sNPF expression in the CC of day 1 5th instar stadium, not of day 3 4th; therefore the stage-specific inhibition of JH synthesis by sNPF was partly due to the stimulative action of AT on the sNPF expression besides the stage-specific expression of the sNPF receptors in the CA, the level of which was high in day 2 4th and day 0 5th instar larvae. The cessation of JH biosynthesis in the last instar larvae is a key event to initiate pupal metamorphosis, and both sNPF and AST-C are key factors in shutting down JH synthesis, along with the decline of ecdysone titer and dopamine.     

Bioindicator potential of Spathosternum prasiniferum prasiniferum (Orthoptera; Acridoidea) in pesticide (azadirachtin)-induced radical toxicity in gonadal/nymphal tissues; correlation with eco-sustainability

Acridids are highly abundant living-organism/major-component in agricultural field, globally. It may act as a dependable bio-indicator species in response to environmental-stress. Antioxidants protect insects by scavenging free radicals. Here, we investigated the dose dependant azadirachtin (C₃₅H₄₄O₁₆ AZT) toxicity on oxidative-biomarker; Alkaline-phosphatase (ALP), malondialdehyde (MDA), non-protein-soluble-thiol (NPSH) and antioxidants like superoxide-dismutase (SOD), catalase (CAT), glutathione-peroxidase (GPx) and amylase of gonads from both sexes and juvenile tissues of common grasshopper, Spathosternum prasiniferum prasiniferum (Walker, 1871). The newly hatched adults (female: male = 1:1) are exposed to 1 to 20 ppm AZT for 48 h and have compared to control and 6 h incubation with same concentration of azadirachtin for in vitro experiments. Both in vivo and in vitro experiment demonstrated significant influences on oxidative biomarkers with increasing antioxidant enzymatic activities in either sex. The male gonads represents decreasing antioxidant enzyme activities compared to female gonads. Lesser protection by CAT and SOD are noticed in male than female in response to AZT exposure. This experiment suggests that azadirachtin increased the major biomarkers with decreasing antioxidant enzyme activities resulting in more free-radicals related threat in adult male gonads. Variable dose responses were noticed on ALP, AchE and MDA in either gender suggesting multiphasic action of the pesticides. Higher mortality rate is noticed in male with lower nymphal life span. Moreover, nymph II^nd is more susceptible than nymph IV^th in vitro intoxication of azadirachtin. Possible life threat of vast representatives of agricultural ecosystem by pesticide should be avoided to maintain different bio-geo cycle and eco-sustainability.     

Methanol-free biosynthesis of fatty acid methyl ester (FAME) in Synechocystis sp. PCC 6803

To meet the increasing global demand of biodiesel over the next decades, alternative methods for producing one of the key constituents of biodiesel (e.g. fatty acid methyl esters (FAMEs)) are needed. Algal biodiesel has been a long-term target compromised by excessive costs for harvesting and processing. In this work, we engineered cyanobacteria to convert carbon dioxide into excreted FAME, without requiring methanol as a methyl donor. To produce FAME, acyl-ACP, a product of the fatty acid biosynthesis pathway, was first converted into free fatty acid (FFA) by a thioesterase, namely ’UcFatB1 from Umbellularia californica. Next, by employing a juvenile hormone acid O-methyltransferase (DmJHAMT) from Drosophila melanogaster and S-adenosylmethionine (SAM) as a methyl donor, FFAs were converted into corresponding FAMEs. The esters were naturally secreted extracellularly, allowing simple product separation by solvent overlay as opposed to conventional algae biodiesel production where the algae biomass must first be harvested and processed for transesterification of extracted triacylglycerols (TAGs). By optimizing both the promoter and RBS elements, up to 120 mg/L of FAMEs were produced in 10 days. Quantification of key proteins and metabolites, together with constructs over-expressing SAM synthetase (MetK), indicated that ’UcFatB1, MetK, and DmJHAMT were the main factors limiting pathway flux. In order to solve the latter limitation, two reconstructed ancestral sequences of DmJHAMT were also tried, resulting in strains showing a broader methyl ester chain-length profile in comparison to the native DmJHAMT. Altogether, this work demonstrates a promising pathway for direct sunlight-driven conversion of CO₂ into excreted FAME.