The design, synthesis, and evaluation of two universal doxorubicin-linkers: preparation of conjugates that retain topoisomerase II activity

The design, synthesis, and evaluation of two N-alkylmaleimide aldehydes have been achieved, which upon reductive alkylation with the C3'-amino group of doxorubicin (DOX) permits the preparation of DOX conjugates via Michael addition of thiol-containing vectors. This method enables the mild, facile, and high-throughput preparation of DOX conjugates that retain the basic C3'-nitrogen, a pre-requisite for topoisomerase II inhibition. Seven DOX-amino acid conjugates were prepared, each displaying similar inhibitory activity as the parent drug.     

Sociosexual behavior, male mating tactics, and the reproductive cycle of giraffe Giraffa camelopardalis

Female distribution exerts a major impact on male mating tactics. Giraffe cows have a reproductive cycle, and a social system, that should favor a male roaming reproductive tactic. We conducted a 2-year study of female Rothschild's giraffe (G. c. rothschildi) reproductive endocrinology in order to characterize attributes of the reproductive cycle and investigate how female endocrine and behavioral cues influence mating activity. We used non-invasive fecal steroid methods to determine reproductive state among females residing in a herd in a large outdoor enclosure. We found that females had an estrous cycle of 14.7 days and that they regularly had multiple ovarian cycles prior to conception. Adult males were more likely to associate with, and sexually investigate, females when they were cycling than when they were either pregnant or acyclic. During the estrous cycle, male-female proximity and sociosexual behavior were more pronounced during the probable fertile phase than the rest of the cycle. Sexual activity between giraffe coincided with the periovulatory period, with male interest in females peaking during the fertile window in the absence of proceptive behavior by females. We conclude that males detect reliable cues revealing female reproductive status and partition their reproductive effort in response to such cues. We propose that male giraffe adopt a roaming reproductive strategy with their large size, enabling them to search for and mate guard fertile females while minimizing metabolic costs.     

Interactions Between Nitrate Assimilation and 2,4-Dinitrophenol Cometabolism in Rhodobacter capsulatus E1F1

The phototrophic, nitrate-photoassimilating bacterium Rhodobacter capsulatus E1F1 cometabolizes 2,4-dinitrophenol (DNP) by photoreducing it to 2-amino-4-nitrophenol under anaerobic conditions. DNP uptake and nitrate metabolism share some biochemical features, and in this article we show that both processes are influenced by each other. Thus, as was demonstrated for nitrate assimilation, DNP uptake requires a thermolabile periplasmic component. Nitrate assimilation is inhibited by DNP, which probably affects the nitrite reduction step because neither nitrate reductase activity nor the transport of nitrate or nitrite is inhibited. On the other hand, DNP uptake is competitively inhibited by nitrate, probably at the transport level, because the nitroreductase activity is not inhibited in vitro by nitrate, nitrite, or ammonium. In addition, the decrease in the intracellular DNP concentration in the presence of nitrate probably inactivates the nitroreductase. These results allow prediction of a negative environmental effect if nitrate and DNP are released together to natural habitats, because it may lead to a lower rate of DNP metabolism and to nitrite accumulation.     

Effects of estradiol on phenylephrine contractility associated with intracellular calcium release in rat aorta

The ability of estradiol to affect phenylephrine-induced contraction and the subsequent increase in resting tone, associated with capacitative Ca²⁺ entry across the plasma membrane, was evaluated in rat aortic rings incubated in Ca²⁺-free solution. The incubation with estradiol (1–100 nM, 5 min) inhibited both the phenylephrine-induced contraction and the IRT. Neither cycloheximide (1 µM; inhibitor of protein synthesis) nor tamoxifen (1 µM; blocker of estrogenic receptors) modified the effects of estradiol. Estradiol (100 µM) also blocked the contractile response to serotonin (10 µM) but not to caffeine (10 mM). In addition, estradiol (100 µM) inhibited the contractile responses to cyclopiazonic acid (1 µM; selective Ca²⁺-ATPase inhibitor) associated with capacitative Ca²⁺ influx through non-L-type Ca²⁺ channels. Finally, estradiol inhibited the Ca²⁺-induced increases in intracellular free Ca²⁺ (after pretreatment with phenylephrine) in cultured rat aorta smooth muscle cells incubated in Ca²⁺-free solution. In conclusion, estradiol interfered in a concentration-dependent manner with Ca²⁺-dependent contractile effects mediated by the stimuli of ₁-adrenergic and serotonergic receptors and inhibited the capacitative Ca²⁺ influx through both L-type and non-L-type Ca²⁺ channels. Such effects are in essence nongenomic and not mediated by the intracellular estrogenic receptor. estrogen; ₁-adrenergic agonists     

Cellular immune responses associated with occult hepatitis C virus infection of the liver

Occult hepatitis C virus (HCV) infection is a type of recently identified chronic infection that is evidenced only by detection of HCV RNA in liver; patients consistently test negative for antibodies to HCV and HCV RNA in serum. Using ex vivo and in vitro measures of T-cell responses, we have identified functional virus-specific memory CD4(+) and CD8(+) T cells in the peripheral blood of patients with occult HCV infection. The features of the virus-specific T cells were consistent with immune surveillance functions, supporting previous exposure to HCV. In addition, the magnitudes of CD4(+) and CD8(+) T-cell responses were in parallel and correlated inversely with the extent of liver HCV infection. The detection of HCV-specific T cells in individuals in whom HCV RNA can persist in the liver despite the absence of viremia and antibodies indicates that HCV replication is prolonged in the face of virus-specific CD4(+) and CD8(+) T-cell responses. These findings demonstrate that HCV-specific cellular immune responses are markers not only of previous exposure to and recovery from HCV but also of ongoing occult HCV infection.     

Seasonal and interannual variation of bacterial production in lowland rivers of the Orinoco basin

1. We examined the influence of hydrologic seasonality on temporal variation of planktonic bacterial production (BP) in relatively undisturbed lowland rivers of the middle Orinoco basin, Venezuela. We sampled two clearwater and two blackwater rivers over 2 years for dissolved organic carbon (DOC), chlorophyll, phosphorus and bacterial abundance to determine their relationship to temporal variation in BP. 2. Dissolved organic carbon concentration was greater in blackwater (543–664 μm ) than in clearwater rivers (184–240 μm ), and was generally higher during periods of rising and high water compared with low water. Chlorophyll concentration peaked (3 μg L^?1) during the first year of study when discharge was lowest, particularly in blackwater rivers. Soluble reactive phosphorus (SRP) was very low in the study rivers (<3.8 μg L^?1) and concentration increased during low water. 3. Average BP was higher in clearwater (0.20–0.26 μg C L^?1 h^?1) than in blackwater rivers (0.14–0.17 μg C L^?1 h^?1), although mean bacterial abundance was similar among rivers (0.6–0.8 × 10⁶ cells mL^?1). 4. Periods of higher chlorophyll a concentration (low water) or flushing of terrestrial organic material (rising water) were accompanied by higher BP, while low BP was observed during the period of high water. 5. Interannual variation in BP was influenced by variations in discharge related to El Niño Southern Oscillation events. 6. Seasonal variation in BP in the study rivers and other tropical systems was relatively small compared with seasonal variation in temperate rivers and lakes. In addition to the low seasonal variation of temperature in the tropics, low overall human disturbance could result in less variation in the inputs of nutrients and carbon to the study rivers compared with more disturbed temperate systems.     

Effect of exogenous arginine on sugarcane ( Saccharum sp.) somatic embryogenesis, free polyamines and the contents of the soluble proteins and proline

The aim of this study was to evaluate the effect of arginine on sugarcane (Saccharum sp.) somatic embryogenesis, free polyamines and other nitrogenous compounds contents. Segments of leaves were used as explants to establish embryogenic cultures on media with 0 and 50.0 mg l⁻¹ arginine. Somatic embryos formation and free polyamines, free proline and total soluble proteins contents were compared. Arginine significantly induced sugarcane somatic embryogenesis. Free proline and protein levels determined in embryogenic cell masses during embryo differentiation-maturation, showed an arginine-induced promotion associated to the enhancement of the embryogenic process. In addition, free putrescine and, in a minor extent, spermidine and spermine contents were enhanced by arginine.     

Potential use of seaweeds in the laying hen ration to improve the quality of n-3 fatty acid enriched eggs

An association between dietary fish oil and decreased yolk weight and reduced sensory quality of eggs has been reported when eggs are enriched with n-3 FA from fish oil. Seaweeds are an important source of compounds that seem to increase egg weight when included in the laying hen diet. The objectives of this study were to determine the influence of the dietary seaweeds Macrocystis pyrifera, Sargassum sinicola and Enteromorpha sp. on the physical quality, lipid composition and consumer acceptability of n-3 FA enriched eggs. One-hundred and forty-four 35-week-old Leghorn hens were randomly distributed in four treatments that consisted of the inclusion of 2% of sardine oil (SO) and 10% of each marine alga (MA) in laying hens' diets; a control diet (C) was also prepared. The study lasted 8 weeks and egg physical quality, egg lipids and sensory attributes were evaluated. The results showed that incorporation of 10% M. pyrifera in the diets is an effective way of increasing the n-3 FA content, the albumen height and yolk color, but not the egg weight, when these are enriched with n-3 FA from fish oil. The egg flavor was also not affected.