Synthesis and in vitro anti-hepatitis B and C virus activities of ring-expanded ('fat') nucleobase analogues containing the imidazo[4,5-e][1,3]diazepine-4,8-dione ring system

As part of our structure–activity relationship studies, we report here the synthesis and in vitro anti-HBV and anti-HCV activities of a number of ring-expanded (‘fat’) nucleobases containing the imidazo[4,5-e][1,3]diazepine-4,8-dione ring system. One of the compounds, ZP-88, exhibited a good activity/toxicity profile against HBV by inhibition of the synthesis of extracellular virion release (EC₅₀ = 1.7 μM, CC₅₀ = 286 μM, SI = 168) and intracellular HBV replication intermediates (EC₅₀ = 8.4 μM, CC₅₀ = 286 μM, SI = 34) in cultured human hepatoblastoma 2.2.15 cells. By contrast, most of the compounds tested against HCV had only marginal activity/toxicity profile, although that was still better than that of the reference compound ribavirin.     

Cadmium biosorption on Spirulina platensis biomass

Dry biomass of Spirulina platensis re-hydrated for 48 h was employed as a biosorbent in tests of cadmium(II) removal from water. Various concentrations of biomass (from 1 to 4 g l⁻¹) and metal (from 100 to 800 mg l⁻¹) were tested. Low biomass levels (X_o  2 g l⁻¹) ensured metal removal up to 98% only at Cd₀= 100 and 200 mg l⁻¹, while X_o  2.0 g l⁻¹ were needed at Cd₀ = 400 mg l⁻¹ to achieve satisfactory results. Whereas X_o = 4.0 g l⁻¹ was effective to remove up to Cd₀ = 500 mg l⁻¹, a further increase in metal concentration (Cd₀ = 600 and 800 mg l⁻¹) led to progressive worsening of the system performance. At a given biomass levels, the kinetics of the process was better at low Cd²⁺ concentrations, while, raising the adsorbent level from 1.0 to 2.0 g l⁻¹ and then to 4.0 g l⁻¹, the rate constant of biosorption increased by about one order of magnitude in both cases and the adsorption capacity of the system progressively decreased from 357 to 149 mg g⁻¹.     

The effect of anoxic treatment on the larvae of six species of dermestids (Coleoptera)

Abstract: Based on surveys of species of museum pest insects commonly found in Finland, Norway, Denmark and Sweden, six species were selected for a study of the effect of exposure to anoxic treatment on the larval stage. An oxygen level of 0.3% (the rest, nitrogen) was applied and lethal exposure times were determined. Anthrenus museorum L. was the most susceptible species (LT₉₉ = 32.2 h), while Attagenus woodroffei Halst. & Green and Attagenus smirnovi Zhantiev (LT₉₉ = 88.1 h) were the most tolerant species. The LT₉₉ value of At. woodroffei was not calculated, as only 50% of the larvae were killed by the treatment. The results indicate that large intraspecific variation is present in these two Attagenus species. Larvae of Anthrenus verbasci (L.) (LT₉₉ = 43.9 h), Reesa vespulae (Milliron) (LT₉₉ = 53.6 h), and Trogoderma angustum (Solier) (LT₉₉ = 57.2 h) showed intermediate tolerance.     

Structure-activity relationships of adenosines with heterocyclic N6-substituents

Two series of N⁶-substituted adenosines with monocyclic and bicyclic N⁶ substituents containing a heteroatom were synthesized in good yields. These derivatives were assessed for their affinity ([³H]CPX), potency, and intrinsic activity (cAMP accumulation) at the A₁ adenosine receptor in DDT₁ MF-2 cells. In the monocyclic series, the N⁶-tetrahydrofuran-3-yl and thiolan-3-yl adenosines (1 and 26, respectively) were found to possess similar activities, whereas the corresponding selenium analogue 27 was found to be more potent. A series of nitrogen containing analogues showed varying properties, N⁶-((3R)-1-benzyloxycarbonylpyrrolidin-3-yl)adenosine (30) was the most potent at the A₁AR; IC₅₀ = 3.2 nM. In the bicyclic series, the effect of a 7-azabicyclo[2.2.1]heptan-2-yl substituent in the N⁶-position was explored. N⁶-(7-Azabicyclo[2.2.1]heptan-2-yl)adenosine (38) proved to be a reasonably potent A₁ agonist (K_i = 51 nM, IC₅₀ = 35 nM) while further substitution on the 7″-nitrogen with tert-butoxycarbonyl (31, IC₅₀ = 2.5 nM) and 2-bromobenzyloxycarbonyl (34, IC₅₀ = 9.0 nM) gave highly potent A₁AR agonists.     

Age, growth and feeding of the common stingray (Dasyatis pastinaca, L., 1758) in the Cilician coastal basin, northeastern Mediterranean Sea

Growth and feeding of stingray ( Dasyatis pastinaca ) were studied using 346 specimens from the Cilician Basin coastal area (northeastern Mediterranean). Age classes between 0 to XII were found. The total length of all specimens ranged from 14.6 to 100.9 cm, and total weight was between 22.5 and 6800 g. Total length (TL)-weight (W) and disc width (DW)-weight (W) relationships were W = 0.0033*L^3.1429 and W = 0.0039*DW^3.4914, respectively. The age data, derived from central readings, were used to estimate the von Bertalanffy length and weight growth parameters. The results were L_∞ = 294.9 cm, W_∞ = 198690.1 g, K = 0.029 year⁻¹; t₀ = −2.2 year. The D. pastinaca diet was composed mainly of crustaceans.     

Biology of the salema, Sarpa salpa (L. 1758) (Pisces, Sparidae) from the middle-eastern Adriatic

Salema Sarpa salpa (Linnaeus, 1758), were caught in the middle-eastern Adriatic Sea from August to November 2004. Length range of the samples was between 10.3 and 43.9 cm, with mean values of 25.5 cm in males, 32.6 cm in females, and 11.4 and 20.2cm in immature and hermaphrodite specimens, respectively. Sex ratio (males : females) was 3.1 : 1. Males were observed up to 37 cm length. Hermaphrodites showed lengths between 18 and 21 cm. The species was characterized by protandric hermaphroditism. Size at sexual maturity was 20.6 cm (2 years old) for males. Total length-total weight relationship for the entire population is described by the parameters a  = 0.00893, and b  = 3.1055. Otolith age readings showed that the population consisted of 15 age groups (1–15 years), including a very high proportion of individuals 1–7 years old. The von Bertalanffy growth parameters were L _∞ = 33.11cm, k  = 0.514, and t ₀ = −0.392 years for males and L _∞ = 40.85cm, k  = 0.179, and t ₀ = −2.606 years for females. Survival rate of females ( S  = 0.870) was much greater than for males ( S  = 0.769).     

The reproduction, age and growth of the spotted rockling

Biology of the Macaronesian endemic rockling Gaidropsarus guttatus was studied in the Azores. The overall sex ratio from the samples was highly in favour of females (1 : 6·33). The growth parameters were L _∞ = 24·23, k  = 1·219 and t ₀ = −0·059. Fish matured at 15 cm L _T and the spawning season was strongly clustered in April.     

Structure-activity studies on different modifications of nociceptin/orphanin FQ: identification of highly potent agonists and antagonists of its receptor

Nociceptin/orphanin FQ (N/OFQ) and its receptor system modulate a variety of biological functions and further understandings of physiological and pathological roles of this system require new potent agonists and antagonists of its receptor. Two series of N/OFQ related analogues were synthesized to investigate the relationship of different modifications. We combined modifications including: (a) Phe⁴→(pF)Phe⁴; (b) Ala⁷, Ala¹¹→Aib⁷, Aib¹¹; (c) Leu¹⁴, Ala¹⁵→Arg¹⁴, Lys¹⁵. Compared with the first series, N-terminus of the second series was changed from Phe¹ to Nphe¹. All the analogues were amidated at C-terminus. These compounds were tested in binding studies on rat brain membranes and mouse vas deferens assay. Results indicated that the compounds of the first series showed higher affinity and potency than N/OFQ (pK_i = 9.33; pEC₅₀ = 7.50). In particular, [(pF)Phe⁴, Aib⁷, Aib¹¹, Arg¹⁴, Lys¹⁵] N/OFQ-NH₂ was found to be a highly potent agonist with pK_i = 10.78 in binding studies and pEC₅₀ = 9.37 in mouse vas deferens assay. The second series all competitively antagonized the effects of N/OFQ in mouse vas deferens assay. [Nphe¹, (pF)Phe⁴, Aib⁷, Aib¹¹, Arg¹⁴, Lys¹⁵] N/OFQ-NH₂ was the best antagonist with pA₂ = 8.39 and showed high binding affinity with pK_i = 9.99. Thus modifications which increase the potency of agonist have synergistic effect on biological activity and a replacement of N-terminus leads to shift of analogues from agonist to antagonist.     

Cytogenetic analysis of Australian tiger beetles (Coleoptera: Cicindelidae): chromosome number, sex-determining system and localization of rDNA genes

The chromosomes of four species of Australian tiger beetle are documented. The male meioformulae of three species of Cicindela (tribe Cicindelini) were determined as follow: Cicindela cardinalba Sumlin 1987, n = 10 + X₁ X₂ X₃Y; C. sp. ( saetigera group) and Cicindela gillesensis Hudson 1994, n = 11 + X₁ X₂ X₃Y. The male meioformula of Megacephala whelani Sumlin 1992 (tribe Megacephalini) is n = 12 + XY. Fluorescence in situ hybridization (FISH) was used to localize the 18S-28S ribosomal gene clusters on male mitotic and meiotic chromosomes and nuclei using a polymerase chain reaction-amplified ribosomal probe. Fluorescence in mitosis and meiotic first prophase stages in the three Cicindela species showed that rDNA genes are in two of the four chromosomes that form the sex vesicle. In M. whelani hybridization in mitosis and first metaphase stages indicate that rDNA genes are in three, medium- to small-sized, autosomal pairs. Silver staining of male meiotic nuclei reveals the presence of active nucleolar organizing regions (NORs) in the sex vesicle of the three species of Cicindela . The cytogenetic and evolutionary implications of these findings are discussed.     

Habitat edges as a potential ecological trap for an insect predator

Abstract.  1. Ecological traps, where animals actively select poor habitat for reproduction over superior habitat, are generally associated with birds at forest edges. This study examines oviposition preference, predation, and parasitism rates in the mantid Stagmomantis limbata to determine the potential generality of this phenomenon.
2. Egg case (oothecae) densities were measured across two edge types (cottonwood and desert scrub) within desert riparian ecosystems. A positive edge effect in oothecae density was found with an approximate three-fold increase in density at cottonwood ( X _edge = 0.05 oothecae/100 m² vs. X _interior = 0.015 oothecae/100 m²) and desert scrub ( X _edge = 0.20 oothecae/100 m² vs. X _interior = 0.06 oothecae/100 m²) edges ( P  < 0.01).
3. Rates of bird predation were significantly higher for oothecae at desert scrub edges and showed a trend of higher predation rates at cottonwood edges, suggesting that riparian habitat edges may be acting as an ecological trap for this mantid species. There was no effect of edges on oothecal parasitism rates.
4. These results provide an example of the effect of habitat edges on a generalist insect predator and indicate that an ecological trap may exist with respect to one of its natural enemies.     

Age and growth analysis of the central mudminnow, Umbra limi (Kirtland, 1840)

A rudimentary understanding of age, growth, and life-span is lacking for many non-game fishes. Growth characteristics of the central mudminnow ( Umbra limi ) have not yet been accurately described using reliable hard part analysis. The utility of scales and otoliths as ageing structures and quantified growth was examined in one lake and one stream population of central mudminnow. Scales were found to be of no utility in determining age due to inconsistent formation of yearly annuli and a high incidence of regenerated scales, while otoliths were easily extracted and considered to be an accurate ageing structure. Ages determined from scales were low compared to those from otoliths, and the difference in age interpreted from the two structures increased with fish age. A power function was fitted to describe the length-weight relationship for this species ( a  = 0.0069, b  = 3.175). Von Bertalanffy growth parameters were estimated and compared for each population (Lake: L _∞ = 114.20 mm, K  = 0.30, t ₀ = −0.93; Stream: L _∞ = 77.59 mm, K  = 0.63, t ₀ = −0.76). The lake population showed greater size at age compared to the stream population, especially at older ages, and achieved a larger maximum size. Growth rate was also greater in the lake population (Lake: 1.74; Stream: 1.09 g year⁻¹). Females were larger at age than males in both populations, however all individuals greater than age 3 were males. This work represents the first successful account of central mudminnow growth using hard part analysis.     

DFT studies of the electronic structure and geometry of 18-crown-6, hexaaza[18]annulene, and their complexes with cations of the heavier alkali and alkaline earth metals

DFT calculations for M@C₁₂H₁₂N₆ and M@C₁₂H₂₄O₆ (where M = Rb⁺, Cs⁺, Sr²⁺, and Ba²⁺, C₁₂H₁₂N₆ = hexaaza[18]annulene, and C₁₂H₂₄O₆ = 18-crown-6) were performed using the recently developed model core potential parametrization. Results show that the ions bind more strongly to C₁₂H₁₂N₆ than to 18-crown-6 moiety; the difference is more pronounced for cations with smaller radii.     

Reproduction, age, growth and mortality of axillary seabream, Pagellus acarne (Sparidae), from the Canarian archipelago

Axillary seabream Pagellus acarne (Risso 1826) caught off the Canary Islands from January 1991 to December 1994 were studied. The length range of the catches was between 11 and 31 cm, with a modal distribution between 17 and 21 cm. The overall ratio of males to females was 1:1.74. Males were observed up to a length of 24 cm. Hermaphrodites were recorded at lengths between 15 and 23 cm. The species was characterized by protandric hermaphroditism. The reproductive season extended from October to March, with a peak in spawning activity in December–January. The size at sexual maturity was 15.8 cm total length (2 years old) for males and 19.4 cm total length (3 years old) for females. The total length–total weight relationship for the entire population is described by the parameters a = 0.0068, and b = 3.2401. Otolith age readings showed that the population exploited consisted of 10 age groups (1–10 years), including a very high proportion of individuals between 1 and 4 years old. The von Bertalanffy growth parameters for all individuals were L_∞ = 32.98 cm, k = 0.22 years⁻¹, and t₀ = −0.87 years. Males grew comparatively slower than females. The instantaneous rates of mortality for all fish were Z = 0.96 years⁻¹, M = 0.30 years⁻¹, and F = 0.66 years⁻¹. The exploitation ratio and the length at first capture were, respectively, E = 0.69 and LC₅₀ = 16.1 cm. The stock is overexploited, therefore measures such as closed seasons or changes in fishing patterns would be desirable to safeguard the spawning stock and recruits.     

Structure and thermal decomposition of methylamine borane

The crystal structure of methylamine borane has been determined and contains parallel chains of dihydrogen-bonded CH₃NH₂BH₃ molecules. Thermal decomposition takes place from the melt (ΔH_fusion = 8.5 kJ mol⁻¹) and begins with the formation of an ionic borohydride. Hydrogen is liberated in two stages, at ca. 100 and 190 °C, with the observed rates during the first stage (ΔH = −25 kJ mol⁻¹, E_a = 115 kJ mol⁻¹) strongly dependent on temperature and time. cis- and trans-N-trimethylcyclotriborazane are formed during the first stage and subsequently cross-link to yield a non-volatile solid. Before this cross-linking, the system exhibited a high degree of volatility, with weight losses in excess of 80% observed in TG experiments using flowing gas.     

Synthesis and characterization of trans-4-(4-chlorophenyl)pyrrolidine-3-carboxamides of piperazinecyclohexanes as ligands for the melanocortin-4 receptor

A series of trans-N-alkyl-4-(4-chlorophenyl)pyrrolidine-3-carboxamides of piperazinecyclohexanemethylamines was synthesized and characterized for binding and function at the melanocortin-4 receptor (MC4R), and several potent benzylamine derivatives were identified. Compound 18v was found to bind MC4R with potent affinity (K_i = 0.5 nM) and high selectivity over the other melanocortin subtypes and behaved as a functional antagonist (IC₅₀ = 48 nM).     

Structural stability of Sclerotium rolfsii ATCC 201126 β-glucan with fermentation time: a chemical, infrared spectroscopic and enzymatic approach

Aims:  Sclerotium rolfsii ATCC 201126 exopolysaccharides (EPSs) recovered at 48 h (EPS I) and 72 h (EPS II) of fermentation, with differences in rheological parameters, hydrogel topography, salt tolerance, antisyneresis, emulsifying and suspending properties, were subjected to a polyphasic characterization in order to detect structural divergences.
Methods and Results:  Fermenter-scale production led to productivity ( P _r) and yield ( Y _P/C) values higher at 48 h ( P _r = 0·542 g l⁻¹ h⁻¹; Y _P/C = 0·74) than at 72 h ( P _r = 0·336 g l⁻¹ h⁻¹; Y _P/C = 0·50). Both EPSs were neutral glucose-homopolysaccharides with a β-(1,3)-glycosidic backbone and single β-(1,6)-glucopyranosyl sidechains regularly attached every three residues in the main chain, as revealed by chemical analyses. The infra-red diagnostic peak at 890 cm⁻¹ confirmed β-glycosidic linkages, while gentiobiose released by β-(1,3)-glucanases confirmed single β-1,6-glycosidic branching for both EPSs.
Conclusions:  The true modular repeating unit of S. rolfsii ATCC 201126 scleroglucan could be resolved. Structural stability was corroborated and no structural differences could be detected as to account for the variations in EPSs behaviour.
Significance and Impact of the Study:  Recovery of S. rolfsii ATCC 201126 scleroglucan at 48 h might be considered based on better fermentation kinetic parameters and no detrimental effects on EPS structural features.     

Endogenous heptapeptide Met-enkephalin-Gly-Tyr binds differentially to duplicate delta opioid receptors from zebrafish

Met-enkephalin-Gly-Tyr (MEGY) is an endogenous peptide that binds to opioid sites in zebrafish and in rat brain homogenates. The aim of this work is to characterize the binding profile of this opioid ligand on two duplicate delta receptors from zebrafish, ZFOR1 and ZFOR4. Our results show that, while ZFOR1 presents one single binding site for [³H]-MEGY (K_D = 4.0 ± 0.4 nM), the experimental data from ZFOR4 fit better to the two-site binding model (K_D1 = 0.8 ± 0.2 nM and K_D2 = 30.2 ± 10.2 nM). Two other MEGY synthetic analogues, (D-Ala²)-MEGY and (D-Ala², Val⁵)-MEGY were also prepared and tested, together with the original peptide MEGY and other opioid ligands, in competition binding assays. While these peptides presented K_i values on the nanomolar range when using [³H]-MEGY as radioligand, these parameters were two orders higher in competition binding assays with the antagonist [³H]-diprenorphine. Functional [³⁵S]GTPγS stimulation analysis has revealed that these two receptors can be activated by several opioid agonists. Our results prove that although the MEGY peptide acts as an agonist on ZFOR1 and ZFOR4, there are subtle pharmacological differences between these two delta opioid receptors from zebrafish.     

Effect of feeding level and feeding frequency on specific dynamic action in Silurus meridionalis

The effect of feeding level ( F _L; 0·5 to 4% dry diet mass per wet fish body mass) and feeding frequency (once every 4 days to twice per day) on postprandial metabolic response was investigated in southern catfish Silurus meridionalis at 27·5° C. The results showed that there was no significant difference in the specific dynamic action (SDA) coefficient among the groups of different feeding levels ( P  > 0·05). The duration increased from 26·0 to 40·0 h and the peak metabolic rate increased from 207·8 to 378·8 mg O₂ kg⁻¹ h⁻¹ when the feeding level was increased from 0·5 to 4%. The relationship between the peak metabolic rate ( R _P, mg O₂ kg⁻¹ h⁻¹) and F _L could be described as: R _P = 175·4 + 47·3 F _L( r ² = 0·943, n  = 40, P  < 0·001). The relationship between the SDA duration ( D , h) and F _L could be described as D =30·97 F _L^0·248 ( r ²=0·729, n =40, P  < 0·001).     

Growth and reproduction of Mesopotamian spiny eel (Mastacembelus mastacembelus Banks & Solander, 1794) in Ataturk Dam Lake (Şanliurfa), Turkey

Age at length, growth and reproduction of 220 Mastacembelus mastacembelus specimens from the Atatürk Dam Lake were studied from July 2005 to July 2006. Total lengths and weights ranged from 7.0 to 85.0 cm and from 6 to 1100 g, respectively. Maximum age was 13 years. The regression model fitted to length and weight data was W  =   0.0228  L ^2.43 for males and W  =   0.0029  L ^2.95 for females. The von Bertalaffy growth equations for males and females were L _t  = 99.2 [1−e^{−0.11 ( t −0.12)}] and L _t  = 69.2 [1−e^{−0.26 ( t −0.35)}], respectively. Males dominated especially at an older age; the overall sex ratio was 1 : 0.63 (M:F). The breeding period was from May to July. Fecundity ranged from 2540 to 24 000 eggs per female.     

N,N′-Ethylene-bis(3-methoxysalicylideneimine) mononuclear (4f) and heterodinuclear (3d–4f) metal complexes: Synthesis, crystal structure and luminescent properties

The stepwise synthesis of mononuclear (4f) and heterodinuclear (3d–4f) Salen-like complexes has been investigated through structural determination of the intermediate and final products occurring in the process. In the first step, reactions of ligand H₂L and Ln(NO₃)₃ · 6H₂O give rise to three mononuclear lanthanide complexes Ln(H₂L)(NO₃)₃ [H₂L = N,N′-ethylene-bis(3-methoxysalicylideneimine), Ln = Nd (1), Eu (2) and Tb (3)], in which N,N′-ethylene-bis(3-methoxysalicylideneimine) acts as tetradentate ligands with the O₂O₂ set of donor atoms capable of effective coordination. These species are fairly stable and have been isolated. Then, addition of Cu(Ac)₂ · H₂O to the mononuclear lanthanide complex yields expected heterodinuclear (3d–4f) complexes Cu(L)Ln(NO₃)₃ · H₂O [Ln = Nd (4) and Eu (5)] where the Cu(II) ion is inserted to the inner N₂O₂ cavity. Luminescent analysis reveals that complex 3 exhibits characteristic metal-centered fluorescence of Tb(III) ion. However, the characteristic luminescence of both Sm(III) and Eu(III) ions is not observed both in solution and solid state of the complexes.