A single amino acid change to Taq DNA polymerase enables faster PCR, reverse transcription and strand-displacement

A change of an aspartic acid to asparagine of Taq

Mutants of Taq DNA polymerase resistant to PCR inhibitors allow DNA amplification from whole blood and crude soil samples

Potent PCR inhibitors in blood and soil samples can cause false negative results from PCR-based clinical and forensic tests. We show that the effect of these inhibitors is primarily upon Taq DNA polymerase, since mutational alteration of the polymerase can overcome the inhibition to the extent that no DNA purification is now required. An N-terminal deletion (Klentaq1) is some 10–100-fold inhibition resistant to whole blood compared to full-length, wild-type (w.t.) Taq, which is strongly inhibited by 0.1–1% blood. Further mutations at codon 708, both in Klentaq 1 and Taq, confer enhanced resistance to various inhibitors of PCR reactions, including whole blood, plasma, hemoglobin, lactoferrin, serum IgG, soil extracts and humic acid, as well as high concentrations of intercalating dyes. Blood PCR inhibitors can predominantly reduce the DNA extension speed of the w.t. Taq polymerase as compared to the mutant enzymes. Single-copy human genomic targets are readily amplified from whole blood or crude soil extract, without pretreatment to purify the template DNA, and the allowed increase in dye concentration overcomes fluorescence background and quenching in real-time PCR of blood

Globular Cluster Population of Hickson Compact Group 22a and 90c

We present the first measurement of the globular cluster populations of galaxies in Hickson compact groups, in order to investigate the effect of these high density environments on the formation and evolution of globular cluster systems. Based on V and R band images that we obtained of HCG 22a and HCG 90c with the ESO New Technology Telescope (NTT), we find a total globular cluster population of $1590\pm 854$ for HCG 22a and $2136\pm 718$ for 90c. The specific frequency for HCG 22a was found to be $S_{N}=1.9\pm 1.0$ and $S_{N}= 3.4\pm 1.1$ for HCG 90c. A power-law fit to the globular cluster radial profile of HCG 22a yields $\sigma\sim R^{-2.01\pm 0.30}$ and for HCG 90c we found $\sigma \sim R^{-1.20\pm0.16}$. A comparison of the globular cluster radial profiles with the surface brightness of the parent galaxy shows that the globular cluster systems are at least as extended as the halo light. The measured values for the specific frequency are consistent with a scenario in which the host galaxies were in a low density ``field-like'' environment when they formed their globular cluster systems.Comment: 28 pages, 13 figures, 3 tables, accepted for publication in the Astrophysical Journa

Physiological Profile of Male Competitive and Recreational Surfers

Surfing consists of both high- and low-intensity paddling of varying durations, using both the aerobic and anaerobic systems. Surf-specific physiological studies lack adequate group sample sizes, and V[Combining Dot Above]O2peak values are yet to determine the differences between competitive and recreational surfers. The purpose of this study was therefore to provide a comprehensive physiological profile of both recreational and competitive surfers. This multisite study involved 62 male surfers, recreational (n = 47) and competitive (n = 15). Anthropometric measurements were conducted followed by dual-energy x-ray absorptiometry, anaerobic testing and finally aerobic testing. V[Combining Dot Above]O2peak was significantly greater in competitive surfers than in recreational surfers (M = 40.71 ± 3.28 vs. 31.25 ± 6.31 ml·kg·min, p \u3c 0.001). This was also paralleled for anaerobic power (M = 303.93 vs. 264.58 W) for competitive surfers. Arm span and lean total muscle mass was significantly (p ≤ 0.01) correlated with key performance variables (V[Combining Dot Above]O2peak and anaerobic power). No significant (p ≥ 0.05) correlations were revealed between season rank and each of the variables of interest (V[Combining Dot Above]O2peak and anaerobic power). Key performance variables (V[Combining Dot Above]O2peak and anaerobic power) are significantly higher in competitive surfers, indicating that this is both an adaptation and requirement in this cohort. This battery of physiological tests could be used as a screening tool to identify an athlete\u27s weaknesses or strengths. Coaches and clinicians could then select appropriate training regimes to address weaknesses

Serum methylarginines and spirometry-measured lung function in older adults

Rationale: Methylarginines are endogenous nitric oxide synthase inhibitors that have been implicated in animal models of lung disease but have not previously been examined for their association with spirometric measures of lung function in humans. Objectives: This study measured serum concentrations of asymmetric and symmetric dimethylarginine in a representative sample of older community-dwelling adults and determined their association with spirometric lung function measures. Methods: Data on clinical, lifestyle, and demographic characteristics, methylated arginines, and L-arginine (measured using LC-MS/MS) were collected from a population-based sample of older Australian adults from the Hunter Community Study. The five key lung function measures included as outcomes were Forced Expiratory Volume in 1 second, Forced Vital Capacity, Forced Expiratory Volume in 1 second to Forced Vital Capacity ratio, Percent Predicted Forced Expiratory Volume in 1 second, and Percent Predicted Forced Vital Capacity. Measurements and Main Results: In adjusted analyses there were statistically significant independent associations between a) higher asymmetric dimethylarginine, lower Forced Expiratory Volume in 1 second and lower Forced Vital Capacity; and b) lower L-arginine/asymmetric dimethylarginine ratio, lower Forced Expiratory Volume in 1 second, lower Percent Predicted Forced Expiratory Volume in 1 second and lower Percent Predicted Forced Vital Capacity. By contrast, no significant associations were observed between symmetric dimethylarginine and lung function. Conclusions: After adjusting for clinical, demographic, biochemical, and pharmacological confounders, higher serum asymmetric dimethylarginine was independently associated with a reduction in key measures of lung function. Further research is needed to determine if methylarginines predict the decline in lung function

Disease prevention strategies for QX disease (Marteilia sydneyi) of Sydney rock oysters (Saccostrea glomerata)

The Sydney rock oyster (Saccostrea glomerata) forms the basis of an important aquaculture industry on the east coast of Australia. During the 1970s, production of S. glomerata began to decline, in part as a result of mortalities arising from Queensland unknown (QX) disease. Histological studies implicated the paramyxean parasite Marteilia sydneyi in the disease outbreaks. Disease zoning was implemented to prevent the spread of M. sydneyi-infected oysters. This control measure hindered rock oyster farming, which historically has relied on transferring wild-caught spat between estuaries for on-growing to market size and has not prevented the subsequent occurrence of QX disease in the Georges and Hawkesbury rivers in central New South Wales. Management of QX disease has been hampered by the complicated life cycle of M. sydneyi, with outbreaks of QX disease likely to be regulated by a combination of the abundance of intermediate host of M. sydneyi, environmental stressors, and the immunocompetence of S. glomerata. The future of the Sydney rock oyster industry relies on understanding these factors and progressing the industry from relying on farming wild-caught seed to the successful commercialization of hatchery-produced QX-resistant S. glomerata

Unanticipated Insights into Biomedicine from the Study of Acupuncture

Research into acupuncture has had ripple effects beyond the field of acupuncture. This paper identifies five exemplars to illustrate that there is tangible evidence of the way insights gleaned from acupuncture research have informed biomedical research, practice, or policy. The first exemplar documents how early research into acupuncture analgesia has expanded into neuroimaging research, broadening physiologic understanding and treatment of chronic pain. The second describes how the acupuncture needle has become a tool to enhance biomedical knowledge of connective tissue. The third exemplar, which illustrates use of a modified acupuncture needle as a sham device, focuses on emergent understanding of placebo effects and, in turn, on insights into therapeutic encounters in treatments unrelated to acupuncture. The fourth exemplar documents that two medical devices now in widespread use were inspired by acupuncture: transcutaneous electrical nerve stimulators for pain control and antinausea wrist bands. The final exemplar describes how pragmatic clinical trial designs applied in acupuncture research have informed current general interest in comparative effectiveness research. In conclusion, these exemplars of unanticipated outcomes of acupuncture research comprise an additional rationale for continued support of basic and clinical research evaluating acupuncture and other under-researched therapies

Physical activity to improve cognition in older adults: can physical activity programs enriched with cognitive challenges enhance the effects? A systematic review and meta-analysis

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