Double Transgenesis of Humanized fat1 and fat2 Genes Promotes Omega-3 Polyunsaturated Fatty Acids Synthesis in a Zebrafish Model

Omega-3 long-chain polyunsaturated fatty acid (n-3 LC-PUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are essential nutrients for human health. However, vertebrates, including humans, have lost the abilities to synthesize EPA and DHA de novo, majorly due to the genetic absence of delta-12 desaturase and omega-3 desaturase genes. Fishes, especially those naturally growing marine fish, are major dietary source of EPA and DHA. Because of the severe decline of marine fishery and the decrease in n-3 LC-PUFA content of farmed fishes, it is highly necessary to develop alternative sources of n-3 LC-PUFA. In the present study, we utilized transgenic technology to generate n-3 LC-PUFA-rich fish by using zebrafish as an animal model. Firstly, fat1 was proved to function efficiently in fish culture cells, which showed an effective conversion of n-6 PUFA to n-3 PUFA with the n-6/n-3 ratio that decreased from 7.7 to 1.1. Secondly, expression of fat1 in transgenic zebrafish increased the 20:5n-3 and 22:6n-3 contents to 1.8- and 2.4-fold, respectively. Third, co-expression of fat2, a fish codon-optimized delta-12 desaturase gene, and fat1 in fish culture cell significantly promoted n-3 PUFA synthesis with the decreased n-6/n-3 ratio from 7.7 to 0.7. Finally, co-expression of fat1 and fat2 in double transgenic zebrafish increased the 20:5n-3 and 22:6n-3 contents to 1.7- and 2.8-fold, respectively. Overall, we generated two types of transgenic zebrafish rich in endogenous n-3 LC-PUFA, fat1 transgenic zebrafish and fat1/fat2 double transgenic zebrafish. Our results demonstrate that application of transgenic technology of humanized fat1 and fat2 in farmed fishes can largely improve the n-3 LC-PUFA production.     

Fatty acids profile and temperature in the cultured marine diatom Odontella aurita

The diatom Odontella aurita has now been industrially cultured and commercialized as a dietary supplement rich in omega-3 fatty acids for several years. In this study, we investigated the effect of three temperatures (8, 16, and 24 °C) on the growth and fatty acid composition of cells harvested during the exponential and stationary growth phases. These temperatures were selected on the basis of photosynthesis responses previously obtained at different temperatures using a modulated fluorometer. Our results confirm that both growth and lipid composition were sensitive to culture temperature. Growth was reduced when O. aurita was cultured at low temperature (8 °C) compared to when it was cultured at high temperatures (16 and 24 °C), but the proportion of polyunsaturated fatty acids (PUFAs, 20:5 n-3 and 22:6 n-3) increased while the level of saturated fatty acids (SFAs, 14:0 and 16:0) decreased in the cells harvested during both the exponential and stationary growth phases. On the other hand, the cells grown at 24 °C displayed a marked decrease in PUFA and an increase in SFA levels. Harvesting time is also a critical parameter in achieving optimum n-3 PUFA productivity during batch cultivation. Indeed, changes in fatty acid composition with growth phase seem to be dependent on the culture temperature, with the most marked effects being observed at 24 °C. PUFA levels (i.e., levels of 20:5 n-3 and 22:6 n-3) increased during the stationary growth phase, while the proportion of SFAs and monounsaturated fatty acids (MUFAs) fell with time. As this species is currently grown in outdoor ponds with seasonal temperature variations (minimal and maximal average temperatures in winter and summer, from 3 to 9 °C and from 13 to 26 °C, respectively), this factor can be expected to have a strong influence on the fatty acid content and composition of the algal biomass harvested and commercialized.     

Common climatic signals affecting oak tree-ring growth in SE Central Europe

A network of 41 local tree-ring chronologies of oak (Quercus petraea and Quercus robur) in Austria, Hungary, Slovenia, Croatia and Serbia (latitudes 45.00–48.00N, longitudes 13.14–21.63E, altitudes 80–800 m a.s.l.) was constructed and used to establish common climatic signals in oak tree rings in the region. Co-variation of residual chronologies could be resumed in 11 significant principal components (PC), explaining 79 % of common variability. Three of them, PC1, PC2 and PC3, made it possible to identify similarities among the sites. PC1, significantly correlated with all 41 chronologies, indicated a common positive response to precipitation in spring and summer (March and June) and a negative response to temperature in spring and summer (April and June). PC2, significantly correlated with 12 chronologies, indicated a common positive response to precipitation especially in spring (May) and a negative one to high summer temperatures (especially in August) with a pronounced north to south gradient. PC3, significantly correlated with ten chronologies, indicated that a warm previous December and warm current September have a positive effect on tree growth, especially in the south-western part of the study area. The obtained climate–growth relationships will help to understand better the variability of oak growth, to fill palaeoclimatic gaps and to improve dendrochronological research in the region.     

SLDP: a Novel Protein Related to Caleosin Is Associated with the Endosymbiotic Symbiodinium Lipid Droplets from Euphyllia glabrescens

Intracellular lipid droplets (LDs) have been proposed to play a key role in the mutualistic endosymbiosis between reef-building corals and the dinoflagellate endosymbiont Symbiodinium spp. This study investigates and identifies LD proteins in Symbiodinium from Euphyllia glabrescens. Discontinuous Percoll gradient centrifugation was used to separate Symbiodinium cells from E. glabrescens tentacles. Furthermore, staining with a fluorescent probe, Nile red, indicated that lipids accumulated in that freshly isolated Symbiodinium cells and lipid analyses further showed polyunsaturated fatty acids (PUFA) was abundant. The stable LDs were purified from endosymbiotic Symbiodinium cells. The structural integrity of the Symbiodinium LDs was maintained via electronegative repulsion and steric hindrance possibly provided by their surface proteins. Protein extracts from the purified LDs revealed a major protein band with a molecular weight of 20 kDa, which was termed Symbiodinium lipid droplet protein (SLDP). Interestingly, immunological cross-recognition analysis revealed that SLDP was detected strongly by the anti-sesame and anti-cycad caleosin antibodies. It was suggested that the stable Symbiodinium LDs were sheltered by this unique structural protein and was suggested that SLDP might be homologous to caleosin to a certain extent.     

Changes of the testes parameters,carcasses and antlers mass of harvested roe deer (Capreolus capreolus) bucks

The research was conducted during the hunting season in Poland from May to September 2012 in which materials of 58 roe deer bucks carcass, antler and testis masses were compared. In addition, the size of testes with epididymides was assessed. Significantly enlarged testes masses were observed in May till July (one-way ANOVA and LSD test). In August and September, the testes underwent abrupt involution. According to the literature and the results of this study, the enlargement of the testes which starts from the end of February, indicates that spring time is a preparatory period for the rut. On the other hand, monthly carcass mass fluctuations were not observed. There is, however, a trend suggesting that heavier roe deer individuals can display larger antler mass.     

Hatching patterns and larval growth of a triplefin from central Chile inferred by otolith microstructure analysis

The subtidal rocky reefs are home to a diverse range of marine animals, including small cryptic fishes, characterised by a bipartite life cycle, with benthic adults and pelagic larval stage that lasts from several days to several months. Using the otolith microstructure analysis, this study determines the hatching and larval growth patterns of the abundant triplefin Helcogrammoides chilensis (Pisces: Tripterygiidae). Fish larvae were collected during September–October 2010 and between July 2012 and April 2013 in nearshore waters (<500 m) of central Chile. Nearshore time series of ichthyoplankton samples showed that large abundance of this species occurs during early austral spring and autumn seasons. Body lengths ranged from 3.11 to 16.57 mm (1–57 days old). Sagittal microincrement analyses estimate that during the main reproductive season, larval growth rates are slow, varying between 0.145 and 0.156 mm day^?1 at a weekly scale. Back-calculated hatch days and circular statistics indicate a major hatch pulse occurring near full moon of the lunar cycle. These results suggest that reproduction occurs coupled with the upwelling season, which reduces the probability of starvation, and hatching occurs during spring tides (full moon), which increases larval dispersion and population connectivity.     

Interannual variations in primary and secondary growth of Nothofagus pumilio and their relationships with climate

Key message

The aim of this study is to evaluate the relationships between primary and secondary growth as well as the influences of climate variations on both types of growth.

Abstract

The relationships between apical (or primary) and radial (or secondary) growth, and climatic influences on both types of growth, were evaluated for Nothofagus pumilio (Nothofagaceae), the dominant subalpine tree in Patagonia. We measured the spacing and number of nodes of annual shoots developed in the period 2001–2010 in 40 N. pumilio trees growing near the upper treeline in the northern Patagonian Andes (41°S). Variations in ring width at the base of each trunk were also recorded. Interannual variations in primary and secondary growth were significantly related to each other, and to several climate variables. Mean temperatures in winter and early spring (June–October) prior to the period of shoot extension were positively associated with both primary and secondary growth. In addition, total summer precipitation (December–March) was positively related to shoot extension, whereas mean summer temperature during the previous growth season (January–March) was directly related to radial growth. These climatic influences on N. pumilio growth may play a major role in regulating the expressions of preformation and neoformation.     

Hexamermis glossinae [Nematoda: Mermithidae] parasitizing tsetse [Diptera: Glossinidae] in Kenya

Glossina pallidipes Austen,G. brevipalpis Newstead andG. austeni Newstead were collected from 5 sites along the south Kenyan coast over a 2 year period. They were dissected and examined for nematodes. Three of the sites yielded tsetse parasitized by juvenile mermithids identified asHexamermis glossinae Poinar et al. Glossina pallidipes andG. brevipalpis are new host records for this parasite, whileG. austeni was captured infrequently and only at a site that failed to yield other parasitized tsetse. Parasite prevalence was low (0.16–0.61 %) and did not differ between male and female hosts. More tsetse than expected by chance harboured nematodes during the long rains season (April–August) than during the short rains (September–November) or dry season (December–March). Early juvenile stages (0.5–2.5 mm long) were recovered mainly from tsetse less than 50 days old, while late juvenile stages (35–85 mm long) were only found in flies older than 30 days. Late stages occurred singly while early ones usually occurred as two or more per host.     

Seasonal variation of fatty acid content in natural microplankton from the Tumpat coastal waters of the South China Sea

In the search for better understanding on the nutritional quality of natural tropical plankton, samples were collected from shallow coastal waters facing the South China Sea during the dry monsoon (May-September) and the wet monsoon (November-April) seasons from March 1993 to July 1994. The total fatty acid content of the predominantly phytoplankton communities (25-200 microns sieve nets) varied four to fivefold with the lowest value occurring during the dry monsoon when blue-green became predominant. Saturated fatty acid content (SAFA), polyunsaturated fatty acid (PUFA) and total omega 3 (sigma omega 3) showed the same seasonal pattern as the total fatty acid with high values in October to December 1993. When species of the dinoflagellate Peridinium and Ceratium were present in considerable amount, the docosahexaenoic acid DHA content was high, especially from March to May 1993. The maximum content of eicosapentaenoic acid EPA, total omega-3 fatty acid, PUFA and sigma omega 3 in phytoplankton occurred during the pre-monsoon period (October and November 1993) when the diatoms were present in large amounts. The larger fraction sample (> 200 microns sieve nets) which consisted predominantly of zooplankton had high amounts of PUFA from September to November 1993.     

Seasonal changes in the content of lipids,fatty acids,and pigments in brown alga <Emphasis Type="Italic">Costaria costata</Emphasis>

We compared the content of total lipids, particular lipid classes, fatty acids (FAs), and pigments in the brown alga Costaria costata [Turn.] Saund (Laminariaceae) in different seasons: during algal active life (April–June) and in the beginning of their degradation (the end of July). Lipids comprised only a small part of algal extractable compounds (10.9–13.4%). Their higher content was observed in May, in the period of alga preparation to sporogenesis. Polar lipids, glyceroglycolipids (GL) and phospholipids (PL), comprised a great part of total lipids in spring months and June, whereas neutral lipids, in July. GL predominated in polar lipids in spring and summer. The highest content of GL was observed in May and PL in April. Storage lipids comprised a great part of lipids in July. The content of sterols was the highest in April and May. In summer months, the content of sterols decreased and remained on unchanged level until thallus degradation. Polyunsaturated FAs (PUFAs) comprised the most part of FAs. Their highest content was observed in April. The main part of PUFAs were those of ω-3 series. In July, the level of these PUFA decreased substantially. The levels of ω-6 PUFAs were approximately similar in spring and summer months. The lowest content of monounsaturated FAs was detected in April; in May and July; the relative content of these FAs increased sharply. We also detected seasonal changes in the content of pigments and the ratios between them.     

Alteration in the zooplankton community of a fly ash treated lake

The zooplankton community in Lake Charles East, Indiana, was sampled from June, 1974 through September, 1977 as part of a lake restoration study. About 1.8 × 10⁴ kg of lime and 1.8 × 10⁶ kg of ponded fly ash were added to the lake during May through August, 1975 to precipitate phosphate and seal the sediments. Annual mean species number (5.2–11.3) and annual mean species diversity (H′, 0.9–1.3) were highest in 1976, the first year after treatment. By the second post-treatment year these variables had returned to pretreatment levels. Prior to treatment Cladocera were dominant during fall and early winter (Sept.–Dec. 1974), with Copepoda dominant in late winter and spring (Jan.–May 1975). After treatment Cladocera were dominant throughout the fall and winter (Nov. 1957–May 1976). Copepoda were again dominant in August 1976. The short term effect of the treatment appeared to be termination of the latter part of the copepod annual cycle through elimination of aestivating copepodites in the summer and increased abundance of Cladocera during the winter and spring immediately following treatment. Community composition one year after treatment was similar to that observed prior to treatment.     

Sons and daughters beyond your control: episodes in the prehistory of the attention deficit/hyperactivity syndrome

Hyperactive and inattentive children have been discussed in both the pedagogic and medical literature since the nineteenth century, and many controversies associated with attention deficit and hyperactivity disorder (ADHD) have been repeatedly analyzed in different contexts. The ‘prehistory’ of the ADHD concept—that is, up to the definition of ADHD in DSM-III and of the corresponding ‘hyperkinetic disorder’ in ICD-9—is outlined, with an emphasis on the literature not previously discussed in English language reviews of the subject.     

Life History of Ilybius fenestratus (Fabricius) (Coleoptera,Dytiscidae) in a Central Norwegian Lake

The life history of Ilybius fenestratus was studied in Målsj?en, a lake in S?r-Tr?ndelag, Central Norway (63°14'N, 10°26'E), during 1971–1972. Adults and larvae were sampled in activity traps every week during March–October and every 2–3 weeks during November–February. Newly-emerged adults were generally found from mid-July to September, and after overwintering in torpidity, probably in the water, they again appeared in the second half of June to August/September. Laying of eggs took place from early/mid July to early August. Larvae occurred in the traps from late July to early May. I. fenestratus was thus found to be a semivoltine summer breeder, with overwintering larvae the first winter and overwintering adults the next. Both adults and larvae lived in large areas of the lake's littoral zone.     

Relative growth and reproductive cycle of the date mussel Lithophaga lithophaga (Linnaeus, 1758) sampled from the Bizerte Bay (Northern Tunisia)

Relative growth and the reproductive cycle of the date mussel Lithophaga lithophaga were studied from September 2002 to October 2003 in the Bizerte Bay (Northern Tunisia). Relationships between shell length and shell width, shell height as well as volume-related variables (shell dry weight, soft tissues dry weight and total wet weight) showed negative allometries in both sexes except for shell width in males (isometry) and shell height in both sexes (positive allometry). The sex ratio was unbalanced, within the length range of 6–48 mm, 49.43 % of the animals were males, 24.39 % females and 26.17 % sexually undifferentiated; within the length range of 49–92 mm, the respective values were 41.19, 52.7 and 6.1 %. Histological investigations as well as analyses of the condition index and the gonadosomatic index (CI and GSI) revealed the presence of a single reproductive cycle per year. Spawning occurred at the end of August and early September and was associated with a decrease in seawater temperature and salinity. A resting phase occurred in winter, coinciding with the lowest water temperatures. Histological examinations of the gonads of a total of 130 specimens revealed only two cases of hermaphroditism. The present study constitutes a useful baseline for a sustainable management of local wild stocks of L. lithophaga.     

REPRODUCTIVE PERIODICITY OF THE SEA URCHIN,TRIPNEUSTES GRATILLA (L.) IN TAIWAN COMPARED WITH OTHER REGIONS

An annual reproductive periodicity of Tripneustes gratilla in northern Taiwan is revealed by both the gonad development stages and the gonad index. The gonads of T. gratilla recovered from spawning in February—March, propagated gametocytes in April—June, came to prematurity in July—August, and matured in September—November. Spawning occurred in October—December. The breeding pattern of T. gratilla over a broad geographical area appears to be related to seasonal changes of sea temperatures.     

Effects of the season and growth stage on the contents of lipids and photosynthetic pigments in brown alga <Emphasis Type="Italic">Undaria pinnatifida</Emphasis>

Seasonal changes in the contents of lipids and photosynthetic pigments (PSP) in the brown alga Undaria pinnatifida (Harvey) Suringar (Phaeophyceae, Alariaceae) on different stages of its growth were studied. Lipids of all plant growth group comprised glyceroglycolipids (GL), phospholipids, and neutral lipids (NL). The ratio between these lipid groups and the content of particular lipids depended on the season and algal growth stage: NL predominated in seedlings; juvenile algae comprised approximately similar amounts of NL and GL; and in adult algae, GL predominated. In winter and spring, algal tissues contained relatively more free sterols than in summer. Total lipid content in seedlings and juvenile algae was higher then in adult plants. Lipid fatty acid (FA) composition was similar on all growth stages, but the content of major components differed; this is mainly related to 18:4 n-3, 20:4 n-6, and 20:5 n-3 acids. The predominant FAs in seedling lipids were saturated FAs, whereas in the lipids of juvenile and adult algae, polyunsaturated FAs predominated.     

The prognostic value of osteopontin expression in non-small cell lung cancer: a meta-analysis

To investigate the association of Osteopontin (OPN) expression in tumor tissue with clinicopathological features of non-small cell lung carcinoma (NSCLC) patients. Publications assessing the clinicopathological characteristics and prognostic significance of OPN in expression NSCLC were identified up to March 2014. A meta-analysis of eligible studies was performed using standard statistical methods to clarify the association between OPN expression and these clinical parameters. A total of eleven studies met the inclusion criteria, and included 1536 cases of NSCLC tumor tissue and 340 cases of normal lung tissue. The OPN expression rate in NSCLC tissue was higher than normal tissue [Odds ratio (OR) 6.427; 95 % confidence interval (CI) 4.689–8.808; P = 0.000]. Simultaneously, we also found that OPN expression was positively associated with stage (OR 0.332; 95 % CI 0.250–0.440; P = 0.000), lymph node metastasis (OR 3.094; 95 % CI 2.295–4.172; P = 0.000), tumor size (tumor size <3 cm vs. ≥3 cm; OR 0.484; 95 % CI 0.303–0.773; P = 0.002) and pathology (OR 0.611; 95 % CI 0.466–0.800; P = 0.000). It was unrelated that OPN expression in NSCLC tissue with and degree of differentiation and other clinical features (P > 0.05). Experimental findings indicate that, OPN plays a crucial role in the development of NSCLC.