Stepwise Solid-Phase Synthesis of Serine-, Tyrosine- and Homoserine-nucleopeptides

Abstract

Hydroxylated amino acids can be introduced in nucleopeptides using the acetyl group for their side chain protection. Base-stable nucleopeptide analogues are obtained if homoserine is used as the linking residue.     

Paracellular absorption in laboratory mice: Molecule size-dependent but low capacity

Water-soluble nutrients are absorbed by the small intestine via transcellular and paracellular processes. The capacity for paracellular absorption seems lower in nonfliers than in fliers, although that conclusion rests largely on a comparison of relatively larger nonflying mammals (> 155 g) and relatively smaller flying birds (< 155 g). We report on paracellular absorption in laboratory mice, the smallest nonflying mammal species studied to date. Using a standard pharmacokinetic technique, we measured the extent of absorption (fractional absorption = f) of inert carbohydrate probes: l-arabinose (M_r = 150.13 Da) and cellobiose (342.3) that are absorbed exclusively by the paracellular route, and 3-O-methyl d-glucose (3OMD-glucose) (M_r = 194) absorbed both paracellularly and transcellularly. f was measured accurately in urine collection trials of 5–10 h duration. Absorption of 3OMD-glucose by mice was essentially complete (f = 0.95 ± 0.07) and much higher than that for l-arabinose (f = 0.21 ± 0.02), indicating that in mice, like other nonflying mammals, > 80% of glucose is absorbed by mediated process(es) rather than the passive, paracellular route. As in all other vertebrates, absorption of cellobiose (f = 0.13 ± 0.02) was even lower than that for l-arabinose, suggesting an equivalent molecular size cut-off for flying and nonflying animals and thus a comparable effective TJ aperture. An important ecological implication is that smaller water-soluble plant secondary metabolites that have been shown to be absorbed by the paracellular path in cell culture, such as phenolics and alkaloids, might be absorbed in substantial amounts by bats and small birds relative to nonflying mammals such as mice.     

Serum Malondialdehyde Correlates with Therapeutic Efficiency of High Activity Antiretroviral Therapies (HAART) in HIV-1 Infected Children

Serum malondialdehyde (MDA) levels are increased in human immunodeficiency virus (HIV)-infected children, as it happens also in infected adult individuals. Introduction of high activity antiretroviral therapy (HAART) has promoted an intense decline in morbidity and mortality of these patients. Here we present data on the effect of HAART on serum MDA of HIV+ children and compare them with levels prior to HAART. MDA levels reflect, as other markers do, the HAART-induced clinical improvement and probably also the pro-oxidant/antioxidant side effects of the different drugs used. The results herein allow the proposal of including serum MDA levels as an additional parameter for the clinical management of HIV+ children.     

Eating catch of the day: the diet of porbeagle shark Lamna nasus (Bonnaterre 1788) based on stomach content analysis,and the interaction with trawl fisheries in the south-western Atlantic (52° S–56° S)

This work examined the diet of the porbeagle shark Lamna nasus in the south-west Atlantic Ocean (SWAO, Argentina, 52° S–56° S) by analysing the stomach content information obtained by scientific observers who sampled specimens captured as by-catch on-board commercial fishing vessels from 2010 to 2020. A total of 148 fishing sites were analysed, in which the estimated catch was composed mainly of hoki Macruronus magellanicus (56.00%) and southern blue whiting Micromesistius australis (33.13%). From 413 porbeagle sharks sampled (292 females and 121 males) ranging from 71 to 241 cm total length (L_T) (mean: 179.76 ± 26.74 cm), 310 (75.06%) contained food in the stomachs. The forage fish were mainly hoki M. magellanicus (23.53%) and southern blue whiting M. australis (19.05%), followed by the Patagonian sprat Sprattus fuegensis (4.48%) and nototheniids (1.4%). Cephalopods and crustaceans accounted for 10% of the diet. The estimated trophic level was 4.35. Generalized linear models revealed that the consumption of hoki M. magellanicus and southern blue whiting M. australis increased with the L_T of the porbeagle shark. Moreover, smaller porbeagle sharks preyed upon both small and large teleost fish, whereas larger porbeagle sharks predated exclusively upon large fish. The diet of porbeagle shark involved interactions with fisheries as it fed upon the fish species that constituted the main catch in the analysed fishing sites, as well as the main catches of the austral trawl fisheries. The ecological role of porbeagle shark observed in the SWAO exposed implications for fisheries management from a multispecies perspective.     

Genetic improvement of spinosad resistance in the biocontrol agent Orius laevigatus

BioControl - Augmentative biological control has experienced a remarkable success, particularly in protected crops. Yet integrated pest management (IPM) still requires corrective treatments to...     

Purification and characterization of a novel alkaline α-<Emphasis Type="SmallCaps">L</Emphasis>-rhamnosidase produced by <Emphasis Type="Italic">Acrostalagmus luteo albus</Emphasis>

Rhamnosidases are enzymes that catalyze the hydrolysis of terminal nonreducing L-rhamnose for the bioconversion of natural or synthetic rhamnosides. They are of great significance in the current biotechnological area, with applications in food and pharmaceutical industrial processes. In this study we isolated and characterized a novel alkaline rhamnosidase from Acrostalagmus luteo albus, an alkali-tolerant soil fungus from Argentina. We also present an efficient, simple, and inexpensive method for purifying the A. luteo albus rhamnosidase and describe the characteristics of the purified enzyme. In the presence of rhamnose as the sole carbon source, this fungus produces a rhamnosidase with a molecular weight of 109 kDa and a pI value of 4.6, as determined by SDS–PAGE and analytical isoelectric focusing, respectively. This enzyme was purified to homogeneity by chromatographic and electrophoretic techniques. Using p-nitrofenil-α-L-rhamnopiranoside as substrate, the enzyme activity showed pH and temperature optima of 8.0 and 55°C, respectively. The enzyme exhibited Michaelis–Menten kinetics, with K _M and V _max values of 3.38 mmol l⁻¹ and 68.5 mmol l⁻¹ min⁻¹, respectively. Neither divalent cations such as Ca²⁺, Mg²⁺, Mn²⁺, and Co²⁺ nor reducing agents such as β-mercaptoethanol and dithiothreitol showed any effect on enzyme activity, whereas this activity was completely inhibited by Zn²⁺ at a concentration of 0.2 mM. This enzyme showed the capacity to hydrolyze some natural rhamnoglucosides such as hesperidin, naringin and quercitrin under alkaline conditions. Based on these results, and mainly due to the high activity of the A. luteo albus rhamnosidase under alkaline conditions, this enzyme should be considered a potential new biocatalyst for industrial applications.     

Characterization of dequalinium as a XIAP antagonist that targets the BIR2 domain

Inhibitor of apoptosis proteins (IAPs) regulate the activity of caspases in apoptosis. The human X chromosome-encoded IAP (XIAP) is one of the more potent members of the IAP family and it has been described as a central regulator of apoptosis. Thus, molecules that inhibit XIAP could offer therapeutic opportunities to treat unwanted apoptosis inhibition. In the present study we have applied the selective optimization of side activities (SOSA) approach to the discovery of XIAP inhibitors. In this sense, we have identified dequalinium hydrochloride (Dq) as an inhibitor of the XIAP/caspase-3 interaction both in vitro and in cellular assays.     

The evolution of mitochondrial genomes in subterranean caviomorph rodents: adaptation against a background of purifying selection

South American tuco-tucos (Ctenomys) and the related coruro (Spalacopus) are two rodent lineages that have independently colonised the subterranean niche. The energetically demanding lifestyles of these species, coupled with the hypoxic atmospheres characteristic of subterranean environments, may have altered the selective regimes on genes encoding proteins related to cellular respiration. Here, we examined the molecular evolution of 13 protein-coding genes in the mitochondrial genome of seven caviomorph rodents, including these two subterranean genera and their above-ground relatives. Using maximum-likelihood and Bayesian approaches, we estimated rates of synonymous (dS) and nonsynonymous (dN) substitutions. We found a significantly higher ω ratio (dN/dS) in subterranean groups as compared to their non-subterranean counterparts in 11 of 13 genes, although no ω ratio was larger than 1. Additionally, we applied a method based on quantitative physicochemical properties to test for positive selection. Amino acid changes implicated in radical structural or functional shifts in the protein property were found to be ubiquitous across the phylogeny, but concentrated in the subterranean lineages. Convergent changes were also found between the subterranean genera used in this study and other mammals adapted to hypoxia. The results of this study suggest a link between niche shifts and weak directional (or episodic) selection at the molecular level against a background of purifying selection.     

A case study of virilizing adrenal tumor in an adolescent female elite tennis player--insight into the use of anabolic steroids in young athletes

A 14-year-old Caucasian girl was referred to the endocrine clinic for evaluation of voice deepening, facial hirsutism, and acne starting 2 years previously. She had been a competitive tennis player since age 7 years, practicing for 4-6 hours daily. On physical examination she was noticed to have a masculine appearance with mild facial acne and moderate hirsutism. Tanner stage was 1 for breast tissue and 5 for pubic hair. Her androgen levels (testosterone, androstenedione, dehydroepiandrosterone sulfate) were extremely elevated. Adrenal ultrasonography revealed a round left 4.6 × 5.3-cm adrenal mass. Laparoscopic left adrenalectomy was performed. The histologic findings were compatible with a benign adrenocortical tumor. Postoperatively, androgen levels dropped to within the normal range. Breast development proceeded normally, menarche occurred 2 months after tumor resection, and menses has been regular since then. Muscle strength of the dominant and nondominant upper and lower extremities was measured 1 month before surgery and 1 year later, using an isokinetic dynamometer (Biodex Systems II, Biodex, Shirley, NY, USA). There was no significant decrease in overall muscle strength after removal of the virilizing tumor and the marked drop in circulating androgens. In addition, the patient maintained her age category, number 1, national tennis ranking. The results suggest that even extremely high levels of tumor-related circulating androgens had no evident effect on muscle strength and competitive performance in a female adolescent tennis player. The lack of beneficial effect on performance in adolescents, combined with the potentially hazardous side effects of anabolic steroids, suggests that teenage athletes should avoid their use.