Enzymes involved in the repair of damaged DNA

The multitude of enzymes responsible for removing damaged nucleotides from DNA in an error-free manner is reviewed. The most direct mechanisms include enzymatically catalyzed photoreversal of cyclobutane dimers and the removal of the O⁶-methylguanine adduct from alkylated DNA by an enzyme whose presence is dependent on adaptation. The direct removal of either damaged purines or pyrimidines or partial removal of photochemically induced diadducts is catalyzed by DNA glycosylases in the absence of phosphodiester bond hydrolysis. Incision of DNA containing apurinic or apyrimidinic sites arising either spontaneously or by the action of DNA glycosylases is catalyzed by specific endonucleases. The incision of DNA containing bulky adducts is attributed to a multigenically controlled uvr system in Escherichia coli. The mechanisms of damaged nucleotide excision and reinsertion of nucleotides are controlled by unique exonuclease functions in either direct or indirect association with DNA polymerases.     

Peripherally inserted central venous catheters. Low-risk alternatives for ongoing venous access.

We prospectively evaluated the use of peripherally inserted central venous catheters to provide ongoing venous access in general medical and surgical patients in a Department of Veterans Affairs medical center. Between 1985 and 1988 trained nurses successfully inserted 393 catheters in 460 suitable patients (an 85.4% success rate). Correct catheter tip placement in the superior vena cava was documented in 359 of the 393 (91.3%) catheter insertions, but an additional 30 catheters were in a position deemed adequate for the intended use. The mean duration of catheter use was 27.6 +/- 5.2 (1 standard deviation) days (median 20 days, range 1 to 370 days). A total of 65 patients left the hospital with catheters in place, with the mean length of catheter use at home being 36.2 +/- 6.0 days (range 2 to 266). In all, 79% of the catheters were in use until the successful completion of therapy or patient death; catheter-related complications led to premature catheter removal in the remaining 21%. Catheter-related complications included bland phlebitis (8.2%), occlusion (8.2%), local infection (3.6%), bacteremia or fungemia (2.1%), mechanical failure or rupture (2.6%), venous thrombosis (0.7%), and other (3.3%). One patient required vein excision for the management of suppurative phlebitis, but no deaths were attributed to catheter use. This study illustrates the use and safety of peripherally inserted central venous catheters to provide reliable vascular access over prolonged periods in an elderly veteran population. At our facility, percutaneous central venous catheters and surgically implanted (Hickman or Broviac) catheters are now reserved for use in patients in whom peripherally inserted catheters cannot be placed.     

Nucleotide sequence of the FokI restriction-modification system: separate strand-specificity domains in the methyltransferase

The genes for FokI, a type-IIS restriction-modification system from Flavobacterium okeanokoites (asymmetric recognition sequence: 5'-GGATG/3'-CCTAC), were cloned into Escherichia coli. Recombinants carrying the fokIR and fokIM genes were found to modify their DNA completely, and to restrict lambdoid phages weakly. The nt sequences of the genes were determined, and the probable start codons were confirmed by aa sequencing. The FokI endonuclease (R · FokI) and methyltransferase (M · FokI are encoded by single, adjacent genes, aligned in the same orientation, in the order M then R. The genes are large by the standards of type-II systems, 1.9 kb for the M gene, and 1.7 kb for the R gene. Preceding each gene is a pair of FokI recognition sites; it is conceivable that interactions between the sites and the FokI proteins could regulate expression of the genes. The aa sequences of the N- and C-terminal halves of M · FokI are similar to one another, and to certain other DNA-adenine methyltransferases, suggesting that the enzyme has a ‘tandem’ structure, such as could have arisen by the fusion of a pair of adjacent, ancestral M genes. Truncated derivatives of M · FokI were constructed by deleting the 5'- or 3' -ends of the fokIM gene. Deleting most of the C-terminus of M · FokI produced derivatives that methylated only the top (GGATG) strand of the recognition sequence. Conversely, deleting most of the N-terminus produced derivatives that methylated only the bottom (CATCC) strand of the recognition sequence. These results indicate that the domains in M · FokI for methylating the two strands of the recognition sequence are largely separate.     

Lack of Influence of Vitamin D Receptor BsmI (rs1544410) Polymorphism on the Rate of Bone Loss in a Cohort of Postmenopausal Spanish Women Affected by Osteoporosis and Followed for Five Years

A longitudinal study was conducted to investigate the relation between a polymorphism in the vitamin D receptor (VDR) gene and changes in bone mineral density (BMD) and quantitative ultrasound of the phalanges (QUS) over a five-year period. The subjects were 456 postmenopausal women with osteoporosis undergoing treatment, aged 59.95±7.97 years (mean±standard deviation [SD]) at baseline. BMD was measured at the hips and lumbar spine by dual-energy X-ray absorptiometry, and QUS was measured by means of amplitude-dependent speed of sound (Ad-SoS) at the phalanges. Lifestyle information was obtained via a questionnaire. The genotype frequencies of the BsmI (rs1544410) gene polymorphism were 29.4%, 47.1%, and 23.5% for bb, Bb, and BB, respectively. After five years, BMD (annual change in %/year) at the femoral neck (FN) showed a significant modification based on the rs1544410 genotype (BB vs Bb); there was an overall decrease in bone mass (-0.70±2.79%/year; P = 0.025). An analysis of covariance with adjustments for age, weight, height, percentage of weight change per year, baseline BMD and calcium intake showed that the observed associations were no longer significant (P = 0.429). No significant associations were found between the QUS measurements and the rs1544410 genotype after the five-year period. Our study limitations includes lack of information about type and length of duration of the osteoporosis treatment. Our results indicate that rs1544410 polymorphisms do not account significantly for the changes in bone mass in Spanish women with osteoporosis undergoing treatment.     

Gender and Age Differences in Hourly and Daily Patterns of Sedentary Time in Older Adults Living in Retirement Communities

Background

Total sedentary time varies across population groups with important health consequences. Patterns of sedentary time accumulation may vary and have differential health risks. The purpose of this study is to describe sedentary patterns of older adults living in retirement communities and illustrate gender and age differences in those patterns.

Methods

Baseline accelerometer data from 307 men and women (mean age = 84±6 years) who wore ActiGraph GT3X+ accelerometers for ≥ 4 days as part of a physical activity intervention were classified into bouts of sedentary time (<100 counts per minute). Linear mixed models were used to account for intra-person and site-level clustering. Daily and hourly summaries were examined in mutually non-exclusive bouts of sedentary time that were 1+, 5+, 10+, 20+, 30+, 40+, 50+, 60+, 90+ and 120+ minutes in duration. Variations by time of day, age and gender were explored.

Results

Men accumulated more sedentary time than women in 1+, 5+, 10+, 20+, 30+, 40+, 50+ and 60+ minute bouts; the largest gender-differences were observed in 10+ and 20+ minute bouts. Age was positively associated with sedentary time, but only in bouts of 10+, 20+, 30+, and 40+ minutes. Women had more daily 1+ minute sedentary bouts than men (71.8 vs. 65.2), indicating they break up sedentary time more often. For men and women, a greater proportion of time was spent being sedentary during later hours of the day than earlier. Gender differences in intra-day sedentary time were observed during morning hours with women accumulating less sedentary time overall and having more 1+ minute bouts.

Conclusions

Patterns identified using bouts of sedentary time revealed gender and age differences in the way in which sedentary time was accumulated by older adults in retirement communities. Awareness of these patterns can help interventionists better target sedentary time and may aid in the identification of health risks associated with sedentary behavior. Future studies should investigate the impact of patterns of sedentary time on healthy aging, disease, and mortality.     

Unidirectional Expiratory Valve Method to Assess Maximal Inspiratory Pressure in Individuals without Artificial Airway

Introduction

Maximal Inspiratory Pressure (MIP) is considered an effective method to estimate strength of inspiratory muscles, but still leads to false positive diagnosis. Although MIP assessment with unidirectional expiratory valve method has been used in patients undergoing mechanical ventilation, no previous studies investigated the application of this method in subjects without artificial airway.

Objectives

This study aimed to compare the MIP values assessed by standard method (MIP_sta) and by unidirectional expiratory valve method (MIP_uni) in subjects with spontaneous breathing without artificial airway. MIP_uni reproducibility was also evaluated.

Methods

This was a crossover design study, and 31 subjects performed MIP_sta and MIP_uni in a random order. MIP_sta measured MIP maintaining negative pressure for at least one second after forceful expiration. MIP_uni evaluated MIP using a unidirectional expiratory valve attached to a face mask and was conducted by two evaluators (A and B) at two moments (Tests 1 and 2) to determine interobserver and intraobserver reproducibility of MIP values. Intraclass correlation coefficient (ICC_[2,1]) was used to determine intraobserver and interobserver reproducibility.

Results

The mean values for MIP_uni were 14.3% higher (-117.3 ± 24.8 cmH₂O) than the mean values for MIP_sta (-102.5 ± 23.9 cmH₂O) (p<0.001). Interobserver reproducibility assessment showed very high correlation for Test 1 (ICC_[2,1] = 0.91), and high correlation for Test 2 (ICC_[2,1] = 0.88). The assessment of the intraobserver reproducibility showed high correlation for evaluator A (ICC_[2,1] = 0.86) and evaluator B (ICC_[2,1] = 0.77).

Conclusions

MIP_uni presented higher values when compared with MIP_sta and proved to be reproducible in subjects with spontaneous breathing without artificial airway.     

Progressive Cognitive Deficit,Motor Impairment and Striatal Pathology in a Transgenic Huntington Disease Monkey Model from Infancy to Adulthood

One of the roadblocks to developing effective therapeutics for Huntington disease (HD) is the lack of animal models that develop progressive clinical traits comparable to those seen in patients. Here we report a longitudinal study that encompasses cognitive and motor assessment, and neuroimaging of a group of transgenic HD and control monkeys from infancy to adulthood. Along with progressive cognitive and motor impairment, neuroimaging revealed a progressive reduction in striatal volume. Magnetic resonance spectroscopy at 48 months of age revealed a decrease of N-acetylaspartate (NAA), further suggesting neuronal damage/loss in the striatum. Postmortem neuropathological analyses revealed significant neuronal loss in the striatum. Our results indicate that HD monkeys share similar disease patterns with HD patients, making them potentially suitable as a preclinical HD animal model.