High-level expression of the Endo-beta-N-acetylglucosaminidase F2 gene in E.coli: one step purification to homogeneity

The Endo F2gene was overexpressed in E.coli as a fusion protein joined to the maltose-binding protein. MBP-Endo F2was found in a highly enriched state as insoluble, inactive inclusion bodies. Extraction of the inclusion bodies with 20% acetic acid followed by exhaustive dialysis rendered the fusion protein active and soluble. MBP-Endo F2was digested with Factor Xaand purified on Q-Sepharose. The enzyme was homogeneous by SDS-PAGE, and appeared as a single symmetrical peak on HPLC. Analysis of the amino-terminus demonstrated conclusively that recombinant Endo F2was homogeneous and identical to the native enzyme.      

The role of lateral roots in bypass flow in rice (Oryza sativa L.)

Although an apoplastic pathway (the so‐called bypass flow) is implicated in the uptake of Na⁺ by rice growing in saline conditions, the point of entry of this flow into roots remains to be elucidated. We investigated the role of lateral roots in bypass flow using the tracer trisodium‐8‐hydroxy‐1,3,6‐pyrenetrisulphonic acid (PTS) and the rice cv. IR36. PTS was identified in the vascular tissue of lateral roots using both epifluorescence microscopy and confocal laser scanning microscopy. Cryo‐scanning electron microscopy and epifluorescence microscopy of sections stained with berberine‐aniline blue revealed that the exodermis is absent in the lateral roots. We conclude that PTS can move freely through the cortical layers of lateral roots, enter the stele and be transported to the shoot via the transpiration stream.     

A novel alpha-D-galactosynthase from Thermotoga maritima converts beta-D-galactopyranosyl azide to alpha-galacto-oligosaccharides

The large-scale production of oligosaccharides is a daunting task, hampering the study of the role of glycans in vivo and the testing of the efficacy of novel glycan-based drugs. Glycosynthases, mutated glycosidases that synthesize oligosaccharides in high yields, are becoming important chemo-enzymatic tools for the production of oligosaccharides. However, while β-glycosynthase can be produced with a rather well-established technology, examples of α-glycosynthases are thus far limited only to enzymes from glycoside hydrolase 29 (GH29), GH31 and GH95 families. α-L-Fucosynthases from GH29 use convenient glycosyl azide derivatives as a strategic alternative to glycosyl fluoride donors. However, the general applicability of this method to other α-glycosynthases is not trivial and remains to be confirmed. Here, β-D-galactopyranosyl azide was converted to α-galacto-oligosaccharides with good yields and high regioselectivity, catalyzed by a novel α-galactosynthase based on the GH36 α-galactosidase from the hyperthermophilic bacterium Thermotoga maritima. These results open a new avenue to the practical synthesis of biologically interesting α-galacto-oligosaccharides and demonstrate more widespread use of β-glycosyl-azide as donors, confirming their utility to expand the repertoire of glycosynthases.     

An investigation into the acute effects of depth jumps on maximal strength performance

Research has demonstrated that high-load low-velocity (HLLV) exercises (≥85% 1 repetition maximum [1RM]) increase performance in subsequent low-load high-velocity (LLHV) exercises, when separated by a rest period ≥4 minutes. To date, few studies have investigated LLHV exercises on subsequent HLLV exercises. The purpose of this study was to compare the effects of 2, 4, or 6 depth jumps (DJs) on subsequent 1RM back squat performance. Fourteen subjects (age 22 ± 4 years, height 177 ± 10 cm, body mass 80.3 ± 14.4 kg) completed five 1RM back squat testing sessions, either control, retest, or 1 of 3 interventions (2, 4, or 6 DJs from a height of 33 cm, 4 minutes before the first 1RM attempt), in a counterbalanced order. Intraclass correlation coefficients demonstrated a high test-retest reliability for the 1RMs (r = 0.989, p < 0.001). Repeated-measures analysis of variance with Bonferroni post hoc analysis revealed significantly greater 1RM performance (140.71 ± 35.68 kg: p = 0.004, 140.50 ± 33.77 kg: p < 0.001, 141.43 ± 34.39 kg: p = 0.002, respectively) for each intervention (2, 4, or 6 repetitions, respectively) compared to the control condition (132.43 ± 34.56 kg). No significant differences were found between interventions (p > 0.05). The findings of this investigation demonstrate that the inclusion of 2, 4, or 6 DJs, 4 minutes before a maximal squat, enhances subsequent strength performance.     

An electromyographical comparison of trunk muscle activity during isometric trunk and dynamic strengthening exercises

The purpose of this study was to compare rectus abdominis and erector spinae muscle activity during isometric (prone bridge [PB] and superman [SM]) and dynamic strengthening exercises (back squat, front squat [FS], and military press). Participants (n = 10, age 21.8 ± 2.6 years; body mass 82.65 ± 10.80 kg, 174.5± 7.2 cm), performed each exercise in a randomized order, using a repeated-measures design. Electromyographical (EMG) activity (sampling at 2,000 Hz) of the rectus abdominis (RA) and the erector spinae (ES) muscles was recorded throughout the duration of the exercises. Intraclass correlations demonstrated the highest levels of reliability for muscle activity during the isometric exercises; however, all exercises demonstrated high level of reliability (r = 0.764-0.998, p ≤ 0.01). The PB demonstrated significantly greater (p < 0.01) RA activity compared to all other exercises. The ES activity was significantly (p < 0.01) greater during the FS (1.010 ± 0.308 root mean square value [RMS (V)]) and SM (0.951 ± 0.217 RMS[V]) and compared to all other exercises, although there was no significant difference (p > 0.05) between the FS and the SM exercise. The PB may be the most suitable exercise for strengthening the RA, compared to dynamic exercises at a low to moderate load, because of a higher level of muscle activity. The FS may be a useful alternative to isometric exercises when strengthening the ES, because it results in slightly higher muscle activity levels when using only a light to moderate load. Because of the dynamic nature of the FS, this may also be more beneficial in transferring to activities of daily living and sporting environments.     

A comparison of maximal squat strength and 5-, 10-, and 20-meter sprint times, in athletes and recreationally trained men

The purpose of this study was to identify whether there was a relationship between relative strength during a 1 repetition maximum (1RM) back squat and 5-, 10-, and 20-m sprint performances in both trained athletes and recreationally trained individuals. Professional rugby league players (n = 24) and recreationally trained individuals (n = 20) participated in this investigation. Twenty-meter sprint time and 1RM back squat strength, using free weights, were assessed on different days. There were no significant (p ≥ 0.05) differences between the well-trained and recreationally trained groups for 5-m sprint times. In contrast, the well-trained group's 10- and 20-m sprint times were significantly quicker (p = 0.004; p = 0.002) (1.78 + 0.06 seconds; 3.03 + 0.09 seconds) compared with the recreationally trained group (1.84 + 0.07 seconds; 3.13 + 0.11 seconds). The athletes were significantly stronger (170.63 + 21.43 kg) than the recreationally trained individuals (135.45 + 30.07 kg) (p = 0.01); however, there were no significant differences (p > 0.05) in relative strength between groups (1.78 + 0.27 kg/kg; 1.78 + 0.33 kg/kg, respectively). Significant negative correlations were found between 5-m sprint time and relative squat strength (r = -0.613, power = 0.96, p = 0.004) and between relative squat strength and 10- and 20-m sprint times in the recreationally trained group (r = -0.621, power = 0.51, p = 0.003; r = -0.604, power = 0.53, p = 0.005, respectively). These results, indicating that relative strength, are important for initial sprint acceleration in all athletes but more strongly related to sprint performance over greater distances in recreationally trained individuals.     

Ambiguity,Ambivalence, and Apprehensions of Taking HIV-1 Pre-Exposure Prophylaxis among Male Couples in San Francisco: A Mixed Methods Study

Objective

We conducted a mixed-methods study to examine serodiscordant and seroconcordant (HIV-positive/HIV-positive) male couples'' PrEP awareness, concerns regarding health care providers offering PrEP to the community, and correlates of PrEP uptake by the HIV-negative member of the couple.

Design

Qualitative sub-study included one-on-one interviews to gain a deeper understanding of participants'' awareness of and experiences with PrEP and concerns regarding health care providers offering PrEP to men who have sex with men (MSM). Quantitative analyses consisted of a cross-sectional study in which participants were asked about the likelihood of PrEP uptake by the HIV-negative member of the couple and level of agreement with health care providers offering PrEP to anyone requesting it.

Methods

We used multivariable regression to examine associations between PrEP questions and covariates of interest and employed an inductive approach to identify key qualitative themes.

Results

Among 328 men (164 couples), 62% had heard about PrEP, but approximately one-quarter were mistaking it with post-exposure prophylaxis. The majority of participants had low endorsement of PrEP uptake and 40% were uncertain if health care providers should offer PrEP to anyone requesting it. Qualitative interviews with 32 men suggest that this uncertainty likely stems from concerns regarding increased risk compensation. Likelihood of future PrEP uptake by the HIV-negative member of the couple was positively associated with unprotected insertive anal intercourse but negatively correlated with unprotected receptive anal intercourse.

Conclusions

Findings suggest that those at greatest risk may not be receptive of PrEP. Those who engage in moderate risk express more interest in PrEP; however, many voice concerns of increased risk behavior in tandem with PrEP use. Results indicate a need for further education of MSM communities and the need to determine appropriate populations in which PrEP can have the highest impact.