The E2F2 Transcription Factor Sustains Hepatic Glycerophospholipid Homeostasis in Mice

Increasing evidence links metabolic signals to cell proliferation, but the molecular wiring that connects the two core machineries remains largely unknown. E2Fs are master regulators of cellular proliferation. We have recently shown that E2F2 activity facilitates the completion of liver regeneration after partial hepatectomy (PH) by regulating the expression of genes required for S-phase entry. Our study also revealed that E2F2 determines the duration of hepatectomy-induced hepatic steatosis. A transcriptomic analysis of normal adult liver identified "lipid metabolism regulation" as a major E2F2 functional target, suggesting that E2F2 has a role in lipid homeostasis. Here we use wild-type (E2F2(+/+)) and E2F2 deficient (E2F2(-/-)) mice to investigate the in vivo role of E2F2 in the composition of liver lipids and fatty acids in two metabolically different contexts: quiescence and 48-h post-PH, when cellular proliferation and anabolic demands are maximal. We show that liver regeneration is accompanied by large triglyceride and protein increases without changes in total phospholipids both in E2F2(+/+) and E2F2(-/-) mice. Remarkably, we found that the phenotype of quiescent liver tissue from E2F2(-/-) mice resembles the phenotype of proliferating E2F2(+/+) liver tissue, characterized by a decreased phosphatidylcholine to phosphatidylethanolamine ratio and a reprogramming of genes involved in generation of choline and ethanolamine derivatives. The diversity of fatty acids in total lipid, triglycerides and phospholipids was essentially preserved on E2F2 loss both in proliferating and non-proliferating liver tissue, although notable exceptions in inflammation-related fatty acids of defined phospholipid classes were detected. Overall, our results indicate that E2F2 activity sustains the hepatic homeostasis of major membrane glycerolipid components while it is dispensable for storage glycerolipid balance.

Causes of fish depletion: a factor analysis approach

A study was conducted in 54 wetlands of 13 districts of Assam, India to evaluate the causes of fish depletion. Twenty-two variables were considered for the study. Seven factors were extracted through factor analysis (Principal Component Analysis) based on Eigen Value Criteria of more than one. These seven factors together accounted for 69.3% of the total variance. Based on the characteristics of the variables, all the factors were given descriptive names. These variables can be used to measure the extent of management deficiency of the causes of fish depletion in the wetlands. The factors are management deficiency, organic load interference, catchment condition, extrinsic influence, fishermen’s ignorance, external environment and aquaculture program. Management deficiency accounted for a substantial portion of the total variance.

Length-weight relationships and relative conditions of two sciaenids Otolithes cuvieri Trewavas and Johnius elongatus Mohan from the Karachi coast

Estimates of length-weight relationship in Otolithes cuvieri justify separate equations for males (log W =-5.0100+3.1365 log L) and females (log W =-5.2000+3.1006 log L). Relative condition factor "Kn" was found to be 0.877-1.946 in males and 0.879-1.328 in females. High "Kn" values during March to September at 180-220 mm TL in either sexes are indicative of the maturation of gonads. Separate equations for length-weight relationship are also justified for males (log W = -5.1126 + 3.0690 log L) and females (log W = -5.6400 + 3.3070 log L) of Johnius elongatus . "Kn" values were found to be 0.924-1.894 for males and 0.894-1.087 for females. High "Kn" values during January-May and August-September at 130 mm TL onward are indicative of gonadal maturation.

ISOLATION AND PROPERTIES OF A BLOOD-COAGULATION FACTOR-X ACTIVATOR FROM THE VENOM OF KING COBRA (OPHIOPHAGUS HANNAH)

A specific blood coagulation factor X activator was purified from the venom of Ophiophagus hannah by gel filtration and two steps of FPLC Mono-Q column ion-exchange chromatography. It showed a single protein band both in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and alkaline polyacrylamide gel electrophoresis. The mol. wt was estimated to be 62,000 in non-reducing conditions and 64,500 in reducing conditions by SDS-PAGE. The isoelectric point was found to be pH 5.6. The enzyme had weak amidolytic activities toward CBS 65-25, but it showed no activities on S-2266, S-2302, thrombin substrate S-2238, plasmin substrate S-2251 or factor Xa substrate S-2222. It had no arginine esterase activity toward substrate benzoylarginine ethylester (BAEE). The enzyme activated factor X in vitro and the effect was absolutely Ca2+ dependent, with a Hill coefficient of 6.83. It could not activate prothrombin nor had any effect on fibrinogen and thus appeared to act specifically on factor X. The procoagulant activity of the enzyme was almost completely inhibited by serine protease inhibitors like PMSF, TPCK and soybean trypsin inhibitor; partially inhibited by L-cysteine. Metal chelator EDTA did not inhibit its procoagulant activity. These results suggest that the factor X activator from O. hannah venom is a serine protease.

Cloning and characterization of a blood coagulation factor IX-binding protein from the venom of Trimeresurus stejnegeri

A blood coagulation factor IX-binding protein (TSV-FIX-BP) was isolated from the snake venom of Trimeresurus stejnegeri. On SDS-polyacrylamide gel electrophoresis, TSV-FIX-BP showed a single band with an apparent molecular weight of 23,000 under non-reducing conditions. and two distinct bands with apparent molecular weights of 14,800 and 14,000 under reducing conditions. cDNA clones containing the coding sequences of TSV-FIX-BP were isolated and sequenced to determine the structure of the precusors of TSV-FIX-BP subunits. The deduced amino acid sequences of two subunits of TSV-FIX-BP were confirmed by N-terminal protein sequencing and trypsin-digested peptide mass fingerprinting. TSV-FIX-BP was a nonenzymatic C-type lectin-like anti-coagulant. The anti-coagulant activity of TSV-FIX-BP was mainly caused by its dose dependent interaction with blood coagulation factor IX but not with blood coagulation factor X. (C) 2003 Elsevier Science Ltd. All rights reserved.

Bm-TFF2, a trefoil factor protein with platelet activation activity from frog Bombina maxima skin secretions

In mammals, trefoil factor family (TFF) proteins are involved in mucosal maintenance and repair, and they are also implicated in tumor suppression and cancer progression. A novel two domain TFF protein from frog Bombina maxima skin secretions (Bm-TFF2) has been purified and cloned. It activated human platelets in a dose-dependent manner and activation of integrin a(11b)beta(3) was involved. Aspirin and apyrase did not largely reduce platelet response to Bm-TFF2 (a 30% inhibition), indicating that the aggregation is not substantially dependent on ADP and thromboxane A2 autocrine feedback. Elimination of external Ca2+ with EGTA did not influence the platelet aggregation induced by Bm-TFF2, meanwhile a strong calcium signal (cytoplasmic Ca2+ release) was detected, suggesting that activation of phospholipase C (PLC) is involved. Subsequent immunoblotting revealed that, unlike in platelets activated by stejnulxin (a glycoprotein VI agonist), PLC gamma 2 was not phosphorylated in platelets activated by Bm-TFF2. FITC-labeled Bm-TFF2 bound to platelet membranes. Bm-TFF2 is the first TFF protein reported to possess human platelet activation activity. (c) 2005 Elsevier Inc. All rights reserved.

Length-weight relationship and relative condition factor in Oreochromis mossambicus (Peters) of Bhima River

Oreochromis mossambicus (Peters), a common freshwater fish of Bhima river, has high economic value and considerable fishery importance. The length-weight relationship in the logarithmic way for this fish can be written as: Log W = - 4.50241627 + 2.884822741 log L. This is close to the cubic law indicating the isometric growth of the fish in its natural habitat. The correlation coefficient (r) was found to be 0.9865 which showed a good relationship between the two parameters. The mean relative condition factor (K sub(n)) was 1.00 suggesting the well-being of the fish.

Length-weight relationship and condition factor of Perna viridis (Linnaeus, 1758) and Meretrix meretrix (Linnaeus, 1758) from Mumbai waters

Length weight relationship of the two commercially important molluscs P. viridis and M. meretrix was found to be W=-0.40263L super(2.044719) & W=-0.04359L super(2.2315498) respectively. Condition factor was recorded to be less than 1.0 for most part of the year in P. viridis and for M. meretrix it ranged from 0.39 to 4.61. The present study reveals that there was allometric growth in both the species and the growth was not satisfactory since it showed lower K-value during most part of the year.

Two Immunoregulatory Peptides with Antioxidant Activity from Tick Salivary Glands

Ticks are blood-feeding arthropods that may secrete immunosuppressant molecules, which inhibit host inflammatory and immune responses and provide survival advantages to pathogens at tick bleeding sites in hosts. In the current work, two families of immunoregulatory peptides, hyalomin-A and -B, were first identified from salivary glands of hard tick Hyalomma asiaticum asiaticum. Three copies of hyalomin-A are encoded by an identical gene and released from the same protein precursor. Both hyalomin-A and -B can exert significant anti-inflammatory functions, either by directly inhibiting host secretion of inflammatory factors such as tumor necrosis factor-alpha, monocyte chemotectic protein-1, and interferon-gamma or by indirectly increasing the secretion of immunosuppressant cytokine of interleukin-10. Hyalomin-A and -B were both found to potently scavenge free radical in vitro in a rapid manner and inhibited adjuvant-induced inflammation in mouse models in vivo. The JNK/SAPK subgroup of the MAPK signaling pathway was involved in such immunoregulatory functions of hyalomin-A and -B. These results showed that immunoregulatory peptides of tick salivary glands suppress host inflammatory response by modulating cytokine secretion and detoxifying reactive oxygen species.

Estimation of RNA/DNA ratio in two north-Indian natural populations of mahseer (Tor tor) and its relationship with growth and hydrobiology

Samples of Tor tor were collected from Bari Reservoir of Udaipur and Narmada River at Hoshangabad (India), in the months of July and November 2005, respectively. Twenty-five samples were collected from each location. Bari Reservoir samples ranged from 17.0 to 24.5 cm in total length and from 75 to 155 g in weight, while Narmada samples ranged from 20.0 to 42.0 cm in length and 90 to 425 g in weight. The nucleic acid content in body muscle of Tor tor and the RNA/DNA ratio were estimated. The age of fishes was estimated by the scale study method and specimens were classified into four age groups. RNA/DNA ratio showed significant linear increase with increase in weight and age till the age of three years after which, the growth rate reduced. The 1-2 year group was the only one common between the two water bodies and a comparison of RNA/DNA ratios showed higher growth rate in Bari Reservoir. The gross primary productivity was also higher in Bari Reservoir being 551 mg cmˉ³ dˉ¹ compared to 404 mg cmˉ³ dˉ¹ observed for Narmada River. The condition factor (K) was found to be higher (1.21) in the fish from the Bari Reservoir compared to those of Narmada River (1.14). The growth rate was higher in females compared to males in >100 g specimens.

Length-weight relationship and relative condition factor in Daysciaena albida (Cuv.) and Gerres filamentosus (Cuv.)

The length-weight relationship of Daysciaena albida and Gerres filamentosus were calculated separately for indeterminants, mature males and mature females. The logarithmic regression equation obtained for D. albida - males: log w = -1.5055 + 2.8618 log l; females: log w = -0.9260 + 2.4089 log l; indeterminants: log w = -l.7188 + 3.0616 log l. The regression co-efficients between males and females, males and in determinants and female and in determinants showed significant differences. In G. filamentosus the relationship can be expressed as males: log w = -1.3224 + 2.8740 log 1; females: log w = -1.2874 + 2.8381 log l; indeterminants: log w = -0.8167 + 2.2558 log l. The difference in regression co-efficients between male and female are insignificant at 5% level whereas significant differences were observed between males and indeterminants and females and indeterminants. The relative condition factor (Kn) was calculated for the above two species. In D. albida the reasons for the fluctuations of Kn values can be attributed to both spawning cycle as well as feeding intensity whereas in G. filamentosus it synchronies mainly with spawning cycle.

Investigation on reproduction process in two species of crab, Eriphia sebana and Ocypode saratan in the intertidal zone of Chabahar area

The present study with headline investigation on reproduction in two species of crab Eriphia sebana and Ocypode saratan was carried out in the intertidal zone of Chabahar in thirteen month from December 2004 to December 2005. Checked samplings have been taken, 45 number Crab monthly from any four stations by manual or use trap. During this study the following subjects were measured: temperature range and salinity, measurable coast granule, determination of sex ratio, relations carapace width with carapace length, Carapace width with total body weight, Gonad weight, gonadosomatic index, condition factor, gastrosomatic index, investigation content in stomach, LM50, growth parameters, plenty distribution length and width and gonad weight and total body weight. Studied on measurable coast granule were expressed that Ocypode saratan in Desalination station, were nest in soils equable sand and this quantity were confirmed in Pozm station. Sex ratio were assign in desalination area and Pozm M: F 0/44:0/56 and in Tiss and Chabahar M:F 0/45:0/55. Carapace length and carapace width (cm) and body weight (g) Furthest were designated in Ocypode saratan within carapace width sequential: In female: 5/42-6/15-105/13 and in male: 5/53-6/25-108/91 and in Eriphia sebana within Tiss area sequential: in female: 5/12-5/94-110/21 and in male 5/14-60/01-114/37. Have been linear relationship between carapace length and carapace width and equaled CW=aCL+b. Weight growth in two species were be modal and equaled BW=aCLb And increased crab weight by built up carapace width. Maximum gonad weight in Ocypode saratan within Desalination area in female have been outcome 3/39 and in male 0/84g and in Eriphia sebana extreme within Tiss during may in female were be 4/18 and in male 1/1g. Stomach content in Eriphia sebana were involved a black until half-purplish liquid and yellowish in Ocypode saratan. Stomach contents identifiable were being in four groups: Molluscoid, Crustacean, Plankton and Fish. Carapace width during the first year of maturation have been LM50:3/77 in Desalination area and LM50:3/92 in Pozm for Ocypode saratan and LM50:4/26 in Tiss and LM50:4/62 in Chabahar. Ability spawning in Eriphia sebana within Tiss has been CW=4/17cm and in Ocypode saratan within Desalination area CW=4/23cm. Maximun value of Loo for Eriphia sebana was equal 59/67 and growth factor K=0/68 within Tiss and Loo =61/64, K=0/65 for Ocypode saratan within Desalination area. Maximun GSI and GI have been within Desalination area and Tiss and minimum within Pozm and Chabahar. The maturity stages of two species were classifed into six stages. Review on GSI, CF have been showed that relation with temperature and salinity and definer in two species have been spawned in two period that Maximun in spring premier than autumn.

Two spawning stocks of icefish Neosalanx taihuensis revealed from annual reproductive cycle analyses

Neosalanx taihuensis were sampled from the Tian-e-zhou Oxbow from March 2006 through November 2007. Two separate spawning seasons were identified based on the annual reproductive cycles of the females, designated as the autumn-spawning season and the spring-spawning season. Lifespan of the offspring of the spring-spawning fish was 1 year, with them dying after the subsequent spring spawning. Autumn-spawned females seem to be the offspring of the spring-spawning fish, based on monthly changes in the length-frequency distributions. Spring-mature females had higher absolute fecundity, gonadosomatic index, and relative condition factor in 2007 than in 2006. Relative condition factor of the immature female offspring of spring-spawning fish was higher in 2007 than in 2006, portending a further increase in reproductive investment during the spring spawning of 2008. The increasing reproductive investment suggests that the population of N. taihuensis in the Tian-e-zhou Oxbow may be recovering from its recent decline.

FLUCTUATION OF SIZE-FRACTIONATED ALKALINE PHOSPHATASE AFTER BLOOM DISAPPEARANCE IN TWO SHALLOW PONDS

The temporal and vertical fluctuations of size fractionated alkaline phosphatase activity (APA) and kinetics parameters as well as orthophosphate (o-P) and chlorophyll concentrations were investigated after bloom disappearance in two shallow ponds A and B from 27 October 2001 to 15 April 2002. Pond A (Microcystis) bloomed seriously but pond B did not. The data of o-P and chlorophyll suggested that phosphorus was the principal limiting nutrimental element and its vertical flux should be regarded as an important driving factor for algal growth. In pond A, the accumulation of algae-derived detritus after bloom disappearance in overlying water stimulated excretion of algal fraction APA, mainly produced by attached bacteria responsible for detritus decomposition, whereas bacterial fraction APA preferred to function in surface water. Interestingly, completely contrary phenomena were observed in pond B. In season, even though no obvious difference for size-fractionated APA in both ponds, the total APA in pond A peaked earlier showing higher activity and efficiency (low K-m and high V-max values) as a result of algal-derived detritus input. In summary, it is suggested that the excretion of alkaline phosphatase with strongly catalyzing efficiency and high activity should be taken as important contributor to algal-derived detritus decomposition, further fueling nutrient recycle and accelerating algal development next year. Furthermore, some inhibitors and surfactants were testified to be good tools to identify the origin of dissolved alkaline phosphatase.

Gene structures and promoter characteristics of interferon regulatory factor 1 (IRF-1), IRF-2 and IRF-7 from snakehead Channa argus

Three interferon regulatory factor (IRF) genes, CaIRF-1, CaIRF-2 and CaIRF-7, and their promoters of snakehead (Channa argus) were cloned and characterized. The CaIRF-1 gene consists of ten exons, spans 4.3 kb and encodes a putative peptide of 299 aa. The CaIRF-2 gene consists of nine exons, spans 8 kb and encodes a putative peptide of 328 aa. The gene organizations of CaIRF-1 and CaIRF-2 are very similar to that of human IRF-1 and IRF-2 except more compact. Comparison of exon-intron organization of the two genes indicated a common evolutionary structure, notably within the exons encoding the DNA binding domain (DBD) of the two factors. The CaIRF-7 gene spans 4.1 kb and encodes a putative peptide of 437 aa. However, the gene organization of CaIRF-7 consisting of ten exons is different to human IRF-7a gene which has an intron in 5' UTR. Three CaIRFs share homology in N-terminal encompassing the DBD that contains a characteristic repeat of tryptophan residues. The promoters of CaIRF-1 and CaIRF-2 genes contain the conserved sites for NF-kappa B and Sp1. The gamma-IFN activation sites (GAS) were found in the promoters of CaIRF-1 and CaIRF-7. The promoter of CaIRF-7 contains conserved interferon stimulating response element (ISRE) which is characteristic of IFN-induced gene promoter, and suggests that there also exist intracellular amplifier circuit in fish IFN signal pathway. Moreover, the element GAAANN oriented in both directions is repeated in CaIRF promoter regions, which confers to further inducibility by IFN. The constitutive expression of CaIRF genes were found to increase obviously in response to induction by the known IFN-inducer poly I:C. (c) 2008 Published by Elsevier Ltd.