Pollination, herbivory, and habitat fragmentation: Their effects on the reproductive fitness of Angadenia berteroi, a native perennial plant of the south Florida pine rocklands

Angadenia berteroi is a tropical perennial subshrub of the pine rocklands with large yellow flowers that set very few fruits. My dissertation seeks to elucidate the factors that affect the reproductive fitness of Angadenia berteroi a native species of the south Florida pine rocklands. I provide novel information on the pollination biology of this native species. I also assess the effects of herbivory on growth and the reproductive success of A. berteroi. Finally, I elucidate how habitat fragmentation and quality are correlated with reproductive fitness of this native perennial plant.^ Using a novel experimental approach, I determined the most effective pollinator group. I used nylon fishing line of widths corresponding to proboscis diameter of the major groups of visitors to examine pollen removal and deposition. In the field, I estimated visitation frequency and efficacy of each pollinator type. Using potted plants, I exposed flowers to single visit from different types of pollinators to measure fruit set. I performed artificial defoliation with scissors on plants growing in the greenhouse to assess the effects of defoliation before flowering as well as during flowering. Additionally, I used structural equation modelling (SEM) to elucidate how A. berteroi reproductive fitness was affected by habitat fragmentation and quality. ^ My experiments provide evidence that Angadenia berteroi is specialized for bee pollination; though butterflies, skippers and others also visit its flowers, A. berteroi is exclusively pollinated by two native bees of the South Florida pine rocklands . This research also demonstrated that herbivory by the oleander moth may have direct and indirect effects on Angadenia berteroi growth and reproductive success. The SEM results suggested that habitat quality (litter depth and subcanopy cover) may favor reproduction in native species of the South Florida pine rocklands that are properly maintained by periodic fires and exotic control. Insights from this threatened and charismatic species may provide impetus to properly manage remaining pine rocklands in South Florida for this and other endemic understory species.^

Sex-Positive Curricula: An Investigation of the Relationship Between Physical Fitness, Self-Concept and Sexual Functioning

Despite the well-recognized benefits of exercise, Americans are gaining weight in astounding proportions and levels of physical activity are on the decline. The purpose of this study was to investigate a relationship between physical fitness, self-concept and sexual health. There is a dearth of knowledge on this relationship specifically in the context of sex-negative curricula, which is the dominate discourse in the United States. One hundred and thirty-three participants between the ages of 18 - 50 volunteered for fitness testing and data collection. Physical fitness was assessed through body fat, resting metabolic rate, cardiovascular endurance, muscular strength, muscular endurance and flexibility. Self-reported exercise was measured using the International Physical Activity Questionnaire. Self-concept was measured by the Six Factor Self-Concept Scale, which presented a total self-concept score and as six individual concepts of self (likability, morality, task accomplishment, giftedness, power and vulnerability). Additionally, sexual function was measured by Derogatis Interview for Sexual Functioning and presented as both an aggregate score and five separate constructs of sexual functioning (fantasy/cognition, arousal, orgasm, behavior/experience, and drive/desire). Questions pertaining to sexual partners, sex education, and demographic information were also included. The results of the General Linear Model indicated significant relationships between physical fitness, self-concept and total sexual functioning. The sexual behavior/experience of men was predicted by body fat percentage and flexibility. In women, behavior/experience was predicted by body fat percentage and arousal was predicted by cardiovascular endurance. Total self-concept was related to muscular endurance. When men were isolated in the analysis, likability was positively related to sexual behavior/experience, and task accomplishment was inversely related to sexual behavior/experience. In women, giftedness was related to cognition/fantasy, arousal, orgasm and total sexual functioning. No relationships were found between physical fitness and the number of sexual partners in men; however, both muscular strength and the power self-concept were significantly related to number of sexual partners in women. As a result of these findings, women may be inclined to exercise to improve arousal and sexual functioning. Furthermore, educators should note the findings of a positive relationship between physical and psychological health and sexual well-being because they provide support for the development and adoption of sex-positive curricula that incorporate potential benefits of sexual activity.

Small-plasmid-mediated antibiotic resistance is enhanced by increases in plasmid copy number and bacterial fitness

Plasmids play a key role in the horizontal spread of antibiotic resistance determinants among bacterial pathogens. When an antibiotic resistance plasmid arrives in a new bacterial host, it produces a fitness cost, causing a competitive disadvantage for the plasmid-bearing bacterium in the absence of antibiotics. On the other hand, in the presence of antibiotics, the plasmid promotes the survival of the clone. The adaptations experienced by plasmid and bacterium in the presence of antibiotics during the first generations of coexistence will be crucial for the progress of the infection and the maintenance of plasmid-mediated resistance once the treatment is over. Here we developed a model system using the human pathogen Haemophilus influenzae carrying the small plasmid pB1000 conferring resistance to β-lactam antibiotics to investigate host and plasmid adaptations in the course of a simulated ampicillin therapy. Our results proved that plasmid-bearing clones compensated for the fitness disadvantage during the first 100 generations of plasmid-host adaptation. In addition, ampicillin treatment was associated with an increase in pB1000 copy number. The augmentation in both bacterial fitness and plasmid copy number gave rise to H. influenzae populations with higher ampicillin resistance levels. In conclusion, we show here that the modulations in bacterial fitness and plasmid copy number help a plasmid-bearing bacterium to adapt during antibiotic therapy, promoting both the survival of the host and the spread of the plasmid.

Fitness cost and interference of Arm/Rmt aminoglycoside resistance with the RsmF housekeeping methyltransferases

Arm/Rmt methyltransferases have emerged recently in pathogenic bacteria as enzymes that confer high-level resistance to 4,6-disubstituted aminoglycosides through methylation of the G1405 residue in the 16S rRNA (like ArmA and RmtA to -E). In prokaryotes, nucleotide methylations are the most common type of rRNA modification, and they are introduced posttranscriptionally by a variety of site-specific housekeeping enzymes to optimize ribosomal function. Here we show that while the aminoglycoside resistance methyltransferase RmtC methylates G1405, it impedes methylation of the housekeeping methyltransferase RsmF at position C1407, a nucleotide that, like G1405, forms part of the aminoglycoside binding pocket of the 16S rRNA. To understand the origin and consequences of this phenomenon, we constructed a series of in-frame knockout and knock-in mutants of Escherichia coli, corresponding to the genotypes rsmF(+), ΔrsmF, rsmF(+) rmtC(+), and ΔrsmF rmtC(+). When analyzed for the antimicrobial resistance pattern, the ΔrsmF bacteria had a decreased susceptibility to aminoglycosides, including 4,6- and 4,5-deoxystreptamine aminoglycosides, showing that the housekeeping methylation at C1407 is involved in intrinsic aminoglycoside susceptibility in E. coli. Competition experiments between the isogenic E. coli strains showed that, contrary to expectation, acquisition of rmtC does not entail a fitness cost for the bacterium. Finally, matrix-assisted laser desorption ionization (MALDI) mass spectrometry allowed us to determine that RmtC methylates the G1405 residue not only in presence but also in the absence of aminoglycoside antibiotics. Thus, the coupling between housekeeping and acquired methyltransferases subverts the methylation architecture of the 16S rRNA but elicits Arm/Rmt methyltransferases to be selected and retained, posing an important threat to the usefulness of aminoglycosides worldwide.

Large-scale screening of a targeted Enterococcus faecalis mutant library identifies envelope fitness factors

Spread of antibiotic resistance among bacteria responsible for nosocomial and community-acquired infections urges for novel therapeutic or prophylactic targets and for innovative pathogen-specific antibacterial compounds. Major challenges are posed by opportunistic pathogens belonging to the low GC% gram-positive bacteria. Among those, Enterococcus faecalis is a leading cause of hospital-acquired infections associated with life-threatening issues and increased hospital costs. To better understand the molecular properties of enterococci that may be required for virulence, and that may explain the emergence of these bacteria in nosocomial infections, we performed the first large-scale functional analysis of E. faecalis V583, the first vancomycin-resistant isolate from a human bloodstream infection. E. faecalis V583 is within the high-risk clonal complex 2 group, which comprises mostly isolates derived from hospital infections worldwide. We conducted broad-range screenings of candidate genes likely involved in host adaptation (e.g., colonization and/or virulence). For this purpose, a library was constructed of targeted insertion mutations in 177 genes encoding putative surface or stress-response factors. Individual mutants were subsequently tested for their i) resistance to oxidative stress, ii) antibiotic resistance, iii) resistance to opsonophagocytosis, iv) adherence to the human colon carcinoma Caco-2 epithelial cells and v) virulence in a surrogate insect model. Our results identified a number of factors that are involved in the interaction between enterococci and their host environments. Their predicted functions highlight the importance of cell envelope glycopolymers in E. faecalis host adaptation. This study provides a valuable genetic database for understanding the steps leading E. faecalis to opportunistic virulence.

Haemophilus influenzae clinical isolates with plasmid pB1000 bearing blaROB-1: fitness cost and interspecies dissemination

Plasmid pB1000 is a mobilizable replicon bearing the bla(ROB-1) beta-lactamase gene that we have recently described in Haemophilus parasuis and Pasteurella multocida animal isolates. Here we report the presence of pB1000 and a derivative plasmid, pB1000', in four Haemophilus influenzae clinical isolates of human origin. Pulsed-field gel electrophoresis showed unrelated patterns in all strains, indicating that the existence of pB1000 in H. influenzae isolates is not the consequence of clonal dissemination. The replicon can be transferred both by transformation and by conjugation into H. influenzae, giving rise to recipients resistant to ampicillin and cefaclor (MICs, > or =64 microg/ml). Stability experiments showed that pB1000 is stable in H. influenzae without antimicrobial pressure for at least 60 generations. Competition experiments between isogenic H. influenzae strains with and without pB1000 revealed a competitive disadvantage of 9% per 10 generations for the transformant versus the recipient. The complete nucleotide sequences of nine pB1000 plasmids from human and animal isolates, as well as the epidemiological data, suggest that animal isolates belonging to the Pasteurellaceae act as an antimicrobial resistance reservoir for H. influenzae. Further, since P. multocida is the only member of this family that can colonize both humans and animals, we propose that P. multocida is the vehicle for the transport of pB1000 between animal- and human-adapted members of the Pasteurellaceae.

Socio-economic status, growth, physical activity and fitness: the Madeira growth study

Within a country social conditions change over time and these conditions vary from country to country. The associations between these conditions, somatic growth, physical activity and fitness reflect these changes. Aim: The study documented variation in somatic growth, physical activity and fitness associated with socio-economic status (SES). Subjects and methods: The study involved 507 subjects (256 boys and 251 girls) from the Madeira Growth Study, a mixed longitudinal study of five cohorts (8, 10, 12, 14 and 16 years of age) followed at yearly intervals over 3 years (1996–1998). A total of 1493 observations were made. Anthropometric measurements included lengths, body mass, skeletal breadths, girths and skinfolds. Physical activity and SES were collected via questionnaire and interview. Physical fitness was assessed using the Eurofit test battery. Variation in somatic growth, physical activity and physical fitness by SES (high, average and low) was tested with analysis of variance. Results: Significant differences between SES groups were observed for height, body mass and skinfolds. Boys and girls from high SES groups were taller, heavier and fatter (subscapular and triceps skinfolds) than their peers from average and low SES groups. At some age intervals, the high SES group had larger skeletal breadths (girls) and girths (boys and girls) than low SES. Small SES differences were observed for physical activity (sport and leisure-time indices). SES was significantly associated with physical fitness. At some age levels, boys from the low SES group performed better for muscular and aerobic endurance whereas girls from the high SES group performed better for power. Conclusion: Considerable variation in somatic growth and physical fitness in association with SES has been demonstrated, but little association was found for physical activity.

Developmental and physical-fitness associations with gross motor coordination problems in peruvian children

he aims of this cross-sectional study were to examine the developmental characteristics (biological maturation and body size) associated with gross motor coordination problems in 5193 Peruvian children (2787 girls) aged 6–14 years from different geographical locations, and to investigate how the probability that children suffer with gross motor coordination problems varies with physical fitness. Children with gross motor coordination problems were more likely to have lower flexibility and explosive strength levels, having adjusted for age, sex, maturation and study site. Older children were more likely to suffer from gross motor coordination problems, as were those with greater body mass index. However, more mature children were less likely to have gross motor coordination problems, although children who live at sea level or at high altitude were more likely to suffer from gross motor coordination problems than children living in the jungle. Our results provide evidence that children and adolescents with lower physical fitness are more likely to have gross motor coordination difficulties. The identification of youths with gross motor coordination problems and providing them with effective intervention programs is an important priority in order to overcome such developmental problems, and help to improve their general health status.

Synonymous co-variation across the E1/E2 gene junction of hepatitis C virus defines virion fitness

Hepatitis C virus is a positive-sense single-stranded RNA virus. The gene junction partitioning the viral glycoproteins E1 and E2 displays concurrent sequence evolution with the 3′-end of E1 highly conserved and the 5′-end of E2 highly heterogeneous. This gene junction is also believed to contain structured RNA elements, with a growing body of evidence suggesting that such structures can act as an additional level of viral replication and transcriptional control. We have previously used ultradeep pyrosequencing to analyze an amplicon library spanning the E1/E2 gene junction from a treatment naïve patient where samples were collected over 10 years of chronic HCV infection. During this timeframe maintenance of an in-frame insertion, recombination and humoral immune targeting of discrete virus sub-populations was reported. In the current study, we present evidence of epistatic evolution across the E1/E2 gene junction and observe the development of co-varying networks of codons set against a background of a complex virome with periodic shifts in population dominance. Overtime, the number of codons actively mutating decreases for all virus groupings. We identify strong synonymous co-variation between codon sites in a group of sequences harbouring a 3 bp in-frame insertion and propose that synonymous mutation acts to stabilize the RNA structural backbone.

Physical exercise minimizes the toxic triad for cancer: Physical inactivity, low fitness, and obesity

The so-called toxic triad of factors linked to cancer, namely obesity, poor cardiorespiratory fitness and physical inactivity, increase the risk of cancer and, when cancer is present, worsen its prognosis. Thus, obesity and a sedentary lifestyle have been linked to an elevated cancer risk whereas regular physical exercise and good cardiorespiratory function (CRF) diminish this risk. Despite genetic risk factors, there is evidence to show that some lifestyle modifications are capable of reducing the incidence of cancer and its associated morbidity and mortality. Regular physical exercise targeted at maintaining body weight within healthy limits and improving CRF will reduce a person's cancer risk and, once diagnosed, will also improve its prognosis, reducing mortality and the risk of disease recurrence through similar effects. In this review, we describe how physical activity can be used as a pleiotropic, coadjuvant tool to minimize the toxic triad for cancer and update the mechanisms proposed to date for the effects observed.

Preparing fitness programmes

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Safety guidelines for group fitness to music

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Analysis of the aerobic-anaerobic transition in elite cyclists during incremental exercise with the use of electromyography

To investigate the validity and reliability of surface electromyography (EMG) as a new non-invasive determinant of the metabolic response to incremental exercise in elite cyclists. The relation between EMG activity and other more conventional methods for analysing the aerobic-anaerobic transition such as blood lactate measurements (lactate threshold (LT) and onset of blood lactate accumulation (OBLA)) and ventilatory parameters (ventilatory thresholds 1 and 2 (VT1 and VT2)) was studied.Twenty eight elite road cyclists (age 24 (4) years; VO2MAX 69.9 (6.4) ml/kg/min; values mean (SD)) were selected as subjects. Each of them performed a ramp protocol (starting at 0 W, with increases of 5 W every 12 seconds) on a cycle ergometer (validity study). In addition, 15 of them performed the same test twice (reliability study). During the tests, data on gas exchange and blood lactate levels were collected to determine VT1, VT2, LT, and OBLA. The root mean squares of EMG signals (rms-EMG) were recorded from both the vastus lateralis and the rectus femoris at each intensity using surface electrodes. Results - A two threshold response was detected in the rms-EMG recordings from both muscles in 90% of subjects, with two breakpoints, EMG(T1) and EMG(T2), at around 60-70% and 80-90% of VO2MAX respectively. The results of the reliability study showed no significant differences (p > 0.05) between mean values of EMG(T1) and EMG(T2) obtained in both tests. Furthermore, no significant differences (p > 0.05) existed between mean values of EMG(T1), in the vastus lateralis and rectus femoris, and VT1 and LT (62.8 (14.5) and 69.0 (6.2) and 64.6 (6.4) and 68.7 (8.2)% of VO2MAX respectively), or between mean values of EMG(T2), in the vastus lateralis and rectus femoris, and VT2 and OBLA (86.9 (9.0) and 88.0 (6.2) and 84.6 (6.5) and 87.7 (6.4)% of VO2MAX respectively). Rms-EMG may be a useful complementary non-invasive method for analysing the aerobic-anaerobic transition (ventilatory and lactate thresholds) in elite cyclists.

The importance of physical fitness in the performance of adequate cardiopulmonary resuscitation

The aim of the present investigation was to evaluate the influence of the physical fitness of a cardiopulmonary resuscitation (CPR) provider on the performance of and physiologic response to CPR. To this end, comparisons were made of sedentary and physically active subjects in terms of CPR performance and physiologic variables. Two study groups were established: group P (n = 14), composed of sedentary, professional CPR rescuers (mean [± SD]; age, 34 ± 6 years; V̇O2max, 32.5 ± 5.5 mL/kg/min), and group Ex (n = 14), composed of physically active, nonexperienced subjects (age, 34 ± 6 years; V̇O2max, 44.5 ± 8.5 mL/kg/min). Each subject was required to perform an 18-min CPR session, which involved manual external cardiac compressions (ECCs) on an electronic teaching mannequin following accepted standard CPR guidelines. Subjects' gas exchange parameters and heart rates (HRs) were monitored throughout the trial. Variables indicating the adequacy of the ECCs (ECC depth and the percentage of incorrect compressions and hand placements) also were determined. Overall CPR performance was similar in both groups. The indicators of ECC adequacy fell within accepted limits (ie, an ECC depth between 38 and 51 mm). However, fatigue prevented four subjects from group P from completing the trial. In contrast, the physiologic responses to CPR differed between groups. The indicators of the intensity of effort during the trial, such as HR or percentage of maximum oxygen uptake (V̇O2max) were higher in group P subjects than group Ex subjects, respectively (HRs at the end of the trial, 139 ± 22 vs 115 ± 17 beats/min, p < 0.01; percentage of V̇O2max after 12 min of CPR, 46.7 ± 9.7% vs 37.2 ± 10.4%, p < 0,05). These results suggest that a certain level of physical fitness may be beneficial to CPR providers to ensure the adequacy of chest compressions performed during relatively long periods of cardiac arrest.

Aerobic power and lean mass are indicators of competitive sprint performance among elite female cross-country skiers

The purpose of this study was to establish the optimal allometric models to predict International Ski Federation’s ski-ranking points for sprint competitions (FISsprint) among elite female cross-country skiers based on maximal oxygen uptake (V̇O2max) and lean mass (LM). Ten elite female cross-country skiers (age: 24.5±2.8 years [mean ± SD]) completed a treadmill roller-skiing test to determine V̇O2max (ie, aerobic power) using the diagonal stride technique, whereas LM (ie, a surrogate indicator of anaerobic capacity) was determined by dual-emission X-ray anthropometry. The subjects’ FISsprint were used as competitive performance measures. Power function modeling was used to predict the skiers’ FISsprint based on V̇O2max, LM, and body mass. The subjects’ test and performance data were as follows: V̇O2max, 4.0±0.3 L min-1; LM, 48.9±4.4 kg; body mass, 64.0±5.2 kg; and FISsprint, 116.4±59.6 points. The following power function models were established for the prediction of FISsprint: 3.91×105 ∙ VO -6.002maxand 6.95×1010 ∙ LM-5.25; these models explained 66% (P=0.0043) and 52% (P=0.019), respectively, of the variance in the FISsprint. Body mass failed to contribute to both models; hence, the models are based on V̇O2max and LM expressed absolutely. The results demonstrate that the physiological variables that reflect aerobic power and anaerobic capacity are important indicators of competitive sprint performance among elite female skiers. To accurately indicate performance capability among elite female skiers, the presented power function models should be used. Skiers whose V̇O2max differs by 1% will differ in their FISsprint by 5.8%, whereas the corresponding 1% difference in LM is related to an FISsprint difference of 5.1%, where both differences are in favor of the skier with higher V̇O2max or LM. It is recommended that coaches use the absolute expression of these variables to monitor skiers’ performance-related training adaptations linked to changes in aerobic power and anaerobic capacity.