Impact of testing strategy on expression of upper-body work capacity and one-repetition maximum prediction after resistance training in college-aged men and women

The purpose of this study was to assess the effect of resistance training on upper-body muscular strength and the expression of work capacity and muscular endurance. In addition, a training-induced change in the relationship between muscular strength and endurance was assessed by testing changes in the accuracy of using endurance repetitions to predict 1 repetition maximum (1RM) bench press before and after training. College-aged men (n = 85) and women (n = 62) completed a 12-week linear periodization resistance training program. Before and after training, the subjects were assessed for 1RM and repetitions to fatigue (RTFs) with a submaximal load. After pretraining 1RM determination, the subjects were randomly assigned to perform RTFs at 65% 1RM (n = 74) or 90% 1RM (n = 73). Pretraining and posttraining RTFs were conducted at the same respective % 1RM. Work capacity was determined from repetition weight × RTF. After training, there was a significant increase in 1RM in both men (～14%) and women (～23%). Posttraining RTF was not different from pretraining RTF at 65 %1RM (18.2 ± 5.1 and 19.0 ± 6.0, respectively) but was significantly reduced in the 90% 1RM group (6.1 ± 3.6 vs. 4.5 ± 2.7, respectively). Likewise, there was a differential effect of training on the expression of work capacity, which increased in the 65 % 1RM group (123 ± 155 kg-reps) but decreased in the 90% 1RM group (-62 ± 208 kg-reps); the effect was independent of gender within each testing group. In conclusion, the changes in muscular strength associated with resistance training produced an increase in work capacity when tested with a 65 % 1RM load without a change in endurance. In contrast, both work capacity and endurance decreased when tested with 90% 1RM. Thus, the impact of strength training on work capacity and muscle endurance is specific to the load at which endurance testing is performed.     

A Whole Recombinant Yeast-Based Therapeutic Vaccine Elicits HBV X,S and Core Specific T Cells in Mice and Activates Human T Cells Recognizing Epitopes Linked to Viral Clearance

Chronic hepatitis B infection (CHB) is characterized by sub-optimal T cell responses to viral antigens. A therapeutic vaccine capable of restoring these immune responses could potentially improve HBsAg seroconversion rates in the setting of direct acting antiviral therapies. A yeast-based immunotherapy (Tarmogen) platform was used to make a vaccine candidate expressing hepatitis B virus (HBV) X, surface (S), and Core antigens (X-S-Core). Murine and human immunogenicity models were used to evaluate the type and magnitude of HBV-Ag specific T cell responses elicited by the vaccine. C57BL/6J, BALB/c, and HLA-A*0201 transgenic mice immunized with yeast expressing X-S-Core showed T cell responses to X, S and Core when evaluated by lymphocyte proliferation assay, ELISpot, intracellular cytokine staining (ICS), or tumor challenge assays. Both CD4⁺ and CD8⁺ T cell responses were observed. Human T cells transduced with HBc18–27 and HBs183–91 specific T cell receptors (TCRs) produced interferon gamma (IFNγ following incubation with X-S-Core-pulsed dendritic cells (DCs). Furthermore, stimulation of peripheral blood mononuclear cells (PBMCs) isolated from CHB patients or from HBV vaccine recipients with autologous DCs pulsed with X-S-Core or a related product (S-Core) resulted in pronounced expansions of HBV Ag-specific T cells possessing a cytolytic phenotype. These data indicate that X-S-Core-expressing yeast elicit functional adaptive immune responses and supports the ongoing evaluation of this therapeutic vaccine in patients with CHB to enhance the induction of HBV-specific T cell responses.     

Effects of single- vs. multiple-set resistance training on maximum strength and body composition in trained postmenopausal women

The purpose of this study was to examine the effect of a single- vs. a multiple-set resistance training protocol in well-trained early postmenopausal women. Subjects (N = 71) were randomly assigned to begin either with 12 weeks of the single-set or 12 weeks of the multiple-set protocol. After another 5 weeks of regenerational resistance training, the subgroup performing the single-set protocol during the first 12 weeks crossed over to the 12-week multiple-set protocol and vice versa. Neither exercise type nor exercise intensity, degree of fatigue, rest periods, speed of movement, training sessions per week, compliance and attendance, or periodization strategy differed between exercise protocols. Body mass, body composition, and 1 repetition maximum (1RM) values for leg press, bench press, rowing, and leg adduction were measured at baseline and after each period. Multiple-set training resulted in significant increases (3.5-5.5%) for all 4 strength measurements, whereas single-set training resulted in significant decreases (-1.1 to -2.0%). Body mass and body composition did not change during the study. The results show that, in pretrained subjects, multiple-set protocols are superior to single-set protocols in increasing maximum strength.     

Interactions between insulins and liver membrane receptors of guinea pig,calf and chicken. Exclusion of a species-specific insulin receptor

Insulin binding experiments were performed with liver plasma membranes from guinea pig, calf and chicken. Bound insulin was separated from free insulin by a simple and rapid centrifugation of membranes through a layer of silicon oil. ¹²⁵I-labeled beef insulin was displaced from receptor sites by unlabelled guinea pig, beef and chicken insulin. The receptors of animals with insulins of different biological activity show similar basic characteristics and affinities to the different insulin molecules and thus are not specialised for the interactions with the homologous insulin molecule. The binding capacity of the membranes for beef insulin seems to be inversely related to the affinity of the homologous insulin to the receptor, guinea pig membranes showing the highest and chicken membranes the lowest receptor concentration     

Role of Mex67-Mtr2 in the Nuclear Export of 40S Pre-Ribosomes

Nuclear export of mRNAs and pre-ribosomal subunits (pre40S and pre60S) is fundamental to all eukaryotes. While genetic approaches in budding yeast have identified bona fide export factors for mRNAs and pre60S subunits, little is known regarding nuclear export of pre40S subunits. The yeast heterodimeric transport receptor Mex67-Mtr2 (TAP-p15 in humans) binds mRNAs and pre60S subunits in the nucleus and facilitates their passage through the nuclear pore complex (NPC) into the cytoplasm by interacting with Phe-Gly (FG)-rich nucleoporins that line its transport channel. By exploiting a combination of genetic, cell-biological, and biochemical approaches, we uncovered an unanticipated role of Mex67-Mtr2 in the nuclear export of 40S pre-ribosomes. We show that recruitment of Mex67-Mtr2 to pre40S subunits requires loops emanating from its NTF2-like domains and that the C-terminal FG-rich nucleoporin interacting UBA-like domain within Mex67 contributes to the transport of pre40S subunits to the cytoplasm. Remarkably, the same loops also recruit Mex67-Mtr2 to pre60S subunits and to the Nup84 complex, the respective interactions crucial for nuclear export of pre60S subunits and mRNAs. Thus Mex67-Mtr2 is a unique transport receptor that employs a common interaction surface to participate in the nuclear export of both pre-ribosomal subunits and mRNAs. Mex67-Mtr2 could engage a regulatory crosstalk among the three major export pathways for optimal cellular growth and proliferation.     

Production of a New Thiopeptide Antibiotic,TP-1161, by a Marine Nocardiopsis Species

Twenty-seven marine sediment- and sponge-derived actinomycetes with a preference for or dependence on seawater for growth were classified at the genus level using molecular taxonomy. Their potential to produce bioactive secondary metabolites was analyzed by PCR screening for genes involved in polyketide and nonribosomal peptide antibiotic synthesis. Using microwell cultures, conditions for the production of antibacterial and antifungal compounds were identified for 15 of the 27 isolates subjected to this screening. Nine of the 15 active extracts were also active against multiresistant Gram-positive bacterial and/or fungal indicator organisms, including vancomycin-resistant Enterococcus faecium and multidrug-resistant Candida albicans. Activity-guided fractionation of fermentation extracts of isolate TFS65-07, showing strong antibacterial activity and classified as a Nocardiopsis species, allowed the identification and purification of the active compound. Structure elucidation revealed this compound to be a new thiopeptide antibiotic with a rare aminoacetone moiety. The in vitro antibacterial activity of this thiopeptide, designated TP-1161, against a panel of bacterial strains was determined.Natural products remain the most prolific source of new antimicrobials, and the chemical diversity of natural compounds is still unmatched by combinatorial chemistry approaches (9, 31). While the latter has been successfully applied for lead optimization, it basically failed to deliver genuinely new pharmacophores, especially in the field of antimicrobials (31), mainly due to limitations in the structural variety of compounds represented in combinatorial libraries.Most of the antibiotics in clinical use today have been developed from compounds isolated from bacteria and fungi, with members of the actinobacteria being the dominant source (34). Traditionally, most of these antimicrobials have been isolated from soil-derived actinomycetes of the genus Streptomyces. However, isolation strategies in recent years have been directed to unexploited environments like marine sources (40). Bioprospecting efforts focusing on the isolation and screening of actinobacteria from ocean habitats (25, 27) have added new biodiversity to the order Actinomycetales and revealed a range of novel natural products of pharmacological value. The existence of marine actinobacterial species physiologically and phylogenetically distinct from their terrestrial relatives is now widely accepted, and new taxonomic groups of marine actinomycetes have been described for at least six different families within the order Actinomycetales (12). Apart from being phylogenetically distinct from their terrestrial relatives, marine isolates have been shown to possess specific physiological adaptations (e.g., to high salinity/osmolarity and pressure) to their maritime surroundings and many were found to produce novel and chemically diverse secondary metabolites (10, 13, 35).Most streptomycetes and other filamentous actinomycetes possess numerous gene clusters for the biosynthesis of secondary metabolites (2, 32), and genome sequence studies have shown that large portions of their genomes are devoted to secondary metabolite biosynthesis. Twenty gene clusters coding for known or predicted secondary metabolites were identified in the 8.7-Mb genome of Streptomyces coelicolor A3(2) (2), and 6.4% of the 8.7-Mb genome of Streptomyces avermitilis is dedicated to gene clusters for secondary metabolite biosynthesis (32). The marine actinomycete Salinispora allocates nearly 10% of its 5.2-Mb genome to 17 diverse biosynthetic loci, including polyketide synthases (PKSs), nonribosomal peptide synthetases (NRPSs), and several hybrid clusters (4, 43). Many medicinally important natural products, including antibacterials and antifungals, are synthesized by these multimodular assembly lines (14), and genome mining for secondary metabolite gene clusters has become a common tool to assess the genetic capability of bacteria to produce novel bioactive compounds. However, even for well-studied model antibiotic producers like S. coelicolor A3(2), discrepancies between the number of known metabolites on the one hand and the number of pathways identified from genomic data on the other hand are tremendous (2). These discrepancies can only be explained by the facts that most gene clusters for secondary metabolites are silenced under standard laboratory cultivation conditions and that an expression or upregulation of these pathways is only triggered in response to certain environmental signals. It has been shown that by cultivating bacteria under a range of conditions, it is possible to obtain products of many of these “orphan” biosynthetic pathways (4). Using the OSMAC (one strain-many compounds) approach, Bode et al. were able to isolate more than 100 compounds comprising 25 structural classes from only six microorganisms (4).In this study, marine sediment-derived actinomycete isolates were analyzed for the production of antimicrobial secondary metabolites by using microwell plate fermentations and a range of media and conditions. This approach led to the isolation of a new thiopeptide antibiotic, designated TP-1161, produced by a marine sediment-derived Nocardiopsis isolate. Here we report the isolation and structural and biological characterization of TP-1161.     

Predicting maximal strength in trained postmenopausal woman

The purpose of this study was to present an equation that accurately predicts 1 repetition maximum (RM) over a wide range of repetitions to fatigue (RTF) for 4 different machine resistance exercises in postmenopausal women. Seventy trained women (age = 57.4 +/- 3.1 years) performed maximal and submaximal repetitions on leg press, bench press, rowing, and leg adduction machines at the conclusion of a 2-year training program. Maximal repetitions were performed on each exercise in the following ranges: 3-5RM, 6-10RM, 11-15RM, and 16-20RM. Special regard was taken to maintain the identical execution of each test (i.e., range of motion, starting angle, speed of movement). One cubic polynomial (w(i) [0.988-0.0000584 r(i)(3) + 0.00190 r(i)(2) + 0.0104 r(i),] where w(i) is the load of measurement I, and r(i) is the number of repetitions) accurately predicted 1RM from RTF with mean absolute differences between actual 1RM and predicted 1RM for the 4 exercises of 1.5-3.1% and with coefficients of variation of <3.3%. Equation accuracy was independent of the exercise type or the number of RTF. Thus, this study supported the validity of RTF to adequately estimate 1RM over a wide range of repetitions and within different exercises in trained, older female subjects.     

Non-FG mediated transport of the large pre-ribosomal subunit through the nuclear pore complex by the mRNA export factor Gle2

Multiple export receptors passage bound pre-ribosomes through nuclear pore complexes (NPCs) by transiently interacting with the Phe-Gly (FG) meshwork of their transport channels. Here, we reveal how the non-FG interacting yeast mRNA export factor Gly-Leu-FG lethal 2 (Gle2) functions in the export of the large pre-ribosomal subunit (pre-60S). Structure-guided studies uncovered conserved platforms used by Gle2 to export pre-60S: an uncharacterized basic patch required to bind pre-60S, and a second surface that makes non-FG contacts with the nucleoporin Nup116. A basic patch mutant of Gle2 is able to function in mRNA export, but not pre-60S export. Thus, Gle2 provides a distinct interaction platform to transport pre-60S to the cytoplasm. Notably, Gle2’s interaction platforms become crucial for pre-60S export when FG-interacting receptors are either not recruited to pre-60S or are impaired. We propose that large complex cargos rely on non-FG as well as FG-interactions for their efficient translocation through the nuclear pore complex channel.     

Polo-like kinase-2 is required for centriole duplication in mammalian cells

Centriole duplication initiates at the G1-to-S transition in mammalian cells and is completed during the S and G2 phases. The localization of a number of protein kinases to the centrosome has revealed the importance of protein phosphorylation in controlling the centriole duplication cycle. Here we show that the human Polo-like kinase 2 (Plk2) is activated near the G1-to-S transition of the cell cycle. Endogenous and overexpressed HA-Plk2 localize with centrosomes, and this interaction is independent of Plk2 kinase activity. In contrast, the kinase activity of Plk2 is required for centriole duplication. Overexpression of a kinase-deficient mutant under S-phase arrest blocks centriole duplication. Downregulation of endogenous Plk2 with small hairpin RNAs interferes with the ability to reduplicate centrioles. Furthermore, centrioles failed to duplicate during the cell cycle of human fibroblasts and U2OS cells after overexpression of a Plk2 dominant-negative mutant. These results show that Plk2 is a physiological centrosomal protein and that its kinase activity is likely to be required for centriole duplication near the G1-to-S phase transition.