Comparison of the COBAS TaqMan HBV test with the COBAS Amplicor monitor test for measurement of hepatitis B virus DNA in serum

Quantitation of low hepatitis B virus (HBV) DNA levels in patients with chronic hepatitis B is important for monitoring natural history of disease and treatment efficacy. This study aimed to compare the quantitation range and analytical sensitivity of the newly developed COBAS TaqMan HBV test (TaqMan test) with the COBAS Amplicor HBV Monitor Test (Amplicor test), using the Eurohep HBV reference plasma and serum samples from patients. Serial dilutions (2.7x10(1)-2.7x10(8) copies/ml) of the Eurohep HBV reference plasma and 50 serum samples from chronic hepatitis B patients were tested by both assays. The TaqMan test could detect seven (2.7x10(2)-2.7x10(8) copies/ml) of eight dilutions of the reference plasma, while the Amplicor test could only detect three of them (2.7x10(3)-2.7x10(5) copies/ml). The HBV DNA values measured by the TaqMan test correlated very well with the theoretical Eurohep standard values (r=0.998, P<0.001). There were good correlations between the HBV DNA levels measured by the two assays on both the Eurohep reference plasma (r=0.993, P<0.001) and serum samples from patients (r=0.904, P<0.001). Compared to the Amplicor test, the TaqMan test had a higher sensitivity (50 vs. 300 copies/ml), shorter assay time (6 vs. 10 hr), and wider dynamic range (8 vs. 3 logs), and was more cost-effective in a clinical setting. These data indicate that the TaqMan test is an excellent tool for HBV DNA quantitation.     

The effects of voluntary contraction effort on quadriceps femoris electromyogram median frequency in humans: a muscle and sex comparison

The purpose of this study was to examine the effect of voluntary contraction efforts on the median frequency (f _med) of the electromyogram (EMG) recorded from the quadriceps femoris muscle in healthy men and women. A group of 30 healthy volunteers (15 men, 15 women) were assessed for EMG activity of the vastus medialis (VM), vastus lateralis (VL), and rectus femoris (RF) muscles during isometric contractions with the knee at 60° flexion. Subjects performed a series of 5 s maximal voluntary isometric contractions that anchored the perceptual range with a "10" on a 10-point scale. Sub-maximal isometric contractions were then separately performed at the following perceived effort levels on the 10-point scale: 1, 2, 3, 4, 5, 6, 7, 8 and 9, in a random order. Subjects were instructed to maintain the contraction at each perceived level of effort for 5 s. The f _med of the three muscles was assessed using a power spectrum analysis performed over 11 consecutive, 512 ms, epochs overlapping each other by half their length during the middle 3 s of each contraction. The f _med for each of the 11 epochs was then determined for each muscle, followed by calculation of the means and normalized coefficients of variation [(standard deviation/mean)×100%] for each contraction. The results demonstrated that the mean f _med of VL was significantly greater than those of the other two muscles, and that f _med of RF was significantly greater than that of VM. The VL muscle demonstrated a significant increase in mean f _med across the contraction efforts, compared to the VM and RF muscles that displayed a significant decrease. The men displayed significantly higher f _med values for the VM muscle than did the women, as well as showing a significantly greater increase across the contraction efforts for the VL muscle. The variability of f _med was shown to be significantly higher for the VM muscle, compared to the VL and RF muscles. The findings of this study suggest that the f _med statistic is most sensitive to contraction intensity efforts for the VL muscle, and that men display significantly higher values for the VL and VM muscles, compared to women. Electronic Publication     

Nitric oxide participates in the intestinal pathology associated with murine syngeneic graft-versus-host disease

Syngeneic graft-versus-host disease (SGVHD) develops following lethal irradiation, reconstitution with syngeneic bone marrow, and treatment with a short course of cyclosporin A (CsA) therapy. The disease is characterized by the development of a T helper cell type 1-like cytokine response [interleukin (IL)-12, interferon-gamma (IFN-gamma), and tumor necrosis factor alpha], and macrophage activation is central to development of the syndrome. It has been shown that nitric oxide (NO) participates significantly in the development of allogeneic GVHD. Studies were initiated to determine if NO participates in the pathology associated with SGVHD. Significant increases in inducible NO synthase (iNOS) mRNA and circulating NO were found in the tissues of SGVHD versus control animals. Treatment of SGVHD animals with the iNOS inhibitor aminoguanidine (AG) reversed the pathology associated with this disease. Furthermore, AG treatment reduced the production of IL-12 and IFN-gamma mRNA in the colons of CsA-treated mice. These studies demonstrate that NO participates in the pathological processes that are associated with the development of murine SGVHD.     

The influence of visual illusions on grasp position

 Visual size illusions have been shown to affect perceived object size but not the aperture of the hand when reaching to those same objects. Thus, vision for perception is said to be dissociated from vision for action. The present study examines the effect of visual-position and visual-shape illusions on both the visually perceived center of an object and the position of a grasp on that object when a balanced lift is required. The results for both experiments show that although the illusions influence both the perceived and the grasped estimates of the center position, the grasp position is more veridical. This partial dissociation is discussed in terms of its implications for streams of visual processing. Received: 17 November 1997 / Accepted: 11 September 1998     

Sleep and daytime sleepiness in retinitis pigmentosa patients

OBJECTIVE: We examined sleep, daytime sleepiness and the ability to stay awake during the day in patients affected with retinitis pigmentosa (RP), to further delineate the role of photoreceptors in the circadian cycle. METHODS: Twelve individuals diagnosed with RP (40 +/- 8 years) And 12 normally sighted healthy individuals (39 +/- 7 years) matched for age, body mass index (BMI) and sex were selected for the study. Participants had their sleep recorded on two consecutive nights and were monitored on the two following days. On the first day, their ability to stay awake and on the second, their sleep propensity were assessed using the Maintenance of Wakefulness Test (MWT) and the Multiple Sleep Latency Test (MSLT), respectively. Self-report measures were obtained using the Pittsburgh Sleep Quality Index (PSQI), the Epworth Sleepiness Scale (ESS), and the Toronto Hospital Alertness Test (THAT). RESULTS: Subjective daytime sleepiness (ESS: 9 +/- 5 vs. 6 +/- 4, P=0.053) and objectively measured sleep propensity (MSLT: 10 +/- 5 vs. 17 +/- 3 min, P < 0.000) were significantly higher in RP patients than controls, whilst their alertness (THAT: 29 +/- 9 vs. 38 +/- 7, P=0.016) and ability to stay awake (MWT: 21 +/- 9 vs. 29 +/- 2 min, P=0.006) were significantly reduced. Retinitis pigmentosa participants had more disturbed nighttime sleep, with significantly more awakenings (arousal index: 14 +/- 8 vs. 8 +/- 6 h, P=0.039), and tended to have less rapid eye movement (REM) sleep (19 +/- 5 vs. 22 +/- 3%, P=0.094). CONCLUSION: Patients with RP have increased daytime sleepiness, reduced alertness and more disturbed nighttime sleep of poorer quality than their normally sighted counterparts, suggesting an influence of photoreceptor degeneration on the circadian cycle.     

Patterns of aneuploidy in stage IV clear cell renal cell carcinoma revealed by comparative genomic hybridization and spectral karyotyping

We report the use of spectral karyotyping (SKY) and comparative genomic hybridization (CGH) to describe the numerous genomic imbalances characteristic of stage IV clear cell renal cell carcinoma (CCRCC). SKY and CGH were performed on 10 cell lines established from nephrectomy specimens, and CGH on uncultured material from five of the primary renal tumors. The mutational status of VHL (3p25) and MET (7q31), genes implicated in renal carcinogenesis, were determined for each case. Each case showed marked aneuploidy, with an average number of copy alterations of 14.6 (+/-2.7) in the primary tumors and 19.3 (+/-4.6) in the cell lines. Both whole-chromosome and chromosome-segment imbalances were noted by CGH: consistent losses or gains included +5q23-->ter (100%), -3p14-->ter (80%), and +7 (70%). All VHL mutations and 83% of the genomic imbalances found in the primary tumors were also found in the cell lines derived from them. SKY showed many complex structural rearrangements that were undetected by conventional banding analysis in these solid tumors. All cases with VHL inactivation had 3p loss and 5q gain related primarily to unbalanced translocations between 3p and 5q. In contrast, gains of chromosome 7 resulted primarily from whole-chromosome gains and were not associated with mutations of MET. SKY and CGH demonstrated that genomic imbalances in advanced RCC were the result of either segregation errors [i.e., whole chromosomal gains and losses (7.8/case)] or chromosomal rearrangements (10.7/case), of which the majority were unbalanced translocations.     

Multiple phenotypic divergence of mammary adenocarcinoma cell clones.

We have shown that, with in vitro passage, subclones derived from clonal cell populations of 13762NF mammary adenocarcinoma undergo phenotypic drift and diversification in their cellular properties. Here we examine whether phenotypic divergence of 13762NF cell clones extends to therapeutic treatments used in eliminating mammary tumors and whether the apparent rates of phenotypic divergence vary for different treatments. Six subclones isolated from low passage clone MTF7 (T11 l; tissue culture passage 11) cells were compared to a similar number of subclones isolated from high passage clone MTF7 (T35; tissue culture passage 35) cells. Subclones derived from clone MTF7 (TI l) were relatively homogeneous (not significantly different) in their inherent sensitivities to ionizing radiation, extrapolation coefficients and quasithreshold dose values (D_o = 1·61–1·99 Gy; n=0·89–3·42; D_q = 0–2·34). When the MTF7 (Tll) subclones were examined for their sensitivities to 45°C hyperthermic treatment, the inherent sensitivities and dose-response curve parameters (D_o = 5·24–10·05 min; n = 1·08–10·47; D_q = 0·78–12·31) were heterogeneous (significantly different). In addition, the MTF7 (T1 l) subclones were heterogeneous (significantly different) in their sensitivities and dose-response curve parameters to 5-fluoro-2-deoxyuridine (FUdR) treatment (slope= –0·70 to –1·59; y-intercept = 1·31 × 10² to 47·80 × 10²). The LD₅₀ values for FUdR ranged from 14–150 nm for the MTF7 (T11) subclones. At high passage MTF7 (T35) subclones were heterogeneous in their dose-response parameters to ionizing radiation (D_o = 1·17–2·05 Gy; n = 0·80–41·18; D_q = 1·79-\24·94), hyperthermia (D_o = 3·57–6·32 min; n = 2·08–13·54; D_q = 3·68–9·30) and FUdR (slope= –0·77 to –0·93; y-intercept = 4·64 × 10² to 8·83 × 10²; LD₅₀ = 50–160nm). The results indicate that clonal cells diverge for distinct phenotypic properties at differing rates to form heterogeneous cell populations with unique sensitivities to various therapeutic treatments.     

Epidemiology of group C rotavirus infection in Western New York women of childbearing age.

Umbilical cord serum samples (380), an average of 10 per month for 3 years (1990 to 1992), were tested by indirect immunofluorescence assay for group C rotavirus immunoglobulin G. Thirty percent were positive, suggesting that approximately one-third of women of childbearing age in western New York have experienced group C rotavirus infection.     

An optimal algorithm for configuring delivery options of a one-dimensional intensity-modulated beam

The problem of generating delivery options for one-dimensional intensity-modulated beams (1D IMBs) arises in intensity-modulated radiation therapy. In this paper, we present an algorithm with the optimal running time, based on the 'rightmost-preference' method, for generating all distinct delivery options for an arbitrary 1D IMB. The previously best known method for generating delivery options for a 1D IMB with N left leaf positions and N right leaf positions is a 'brute-force' solution, which first generates all N! possible combinations of the left and right leaf positions and then removes combinations that are not physically allowed delivery options. Compared with the brute-force method, our algorithm has several advantages: (1) our algorithm runs in an optimal time that is linearly proportional to the total number of distinct delivery options that it actually produces. Note that for a 1D IMB with multiple peaks, the total number of distinct delivery options in general tends to be considerably smaller than the worst case N!. (2) Our algorithm can be adapted to generating delivery options subject to additional constraints such as the 'minimum leaf separation' constraint. (3) Our algorithm can also be used to generate random subsets of delivery options; this feature is especially useful when the 1D IMBs in question have too many delivery options for a computer to store and process. The key idea of our method is that we impose an order on how left leaf positions should be paired with right leaf positions. Experiments indicated that our rightmost-preference algorithm runs dramatically faster than the brute-force algorithm. This implies that our algorithm can handle 1D IMBs whose sizes are substantially larger than those handled by the brute-force method. Applications of our algorithm in therapeutic techniques such as intensity-modulated arc therapy and 2D modulations are also discussed.     

Specificity and mechanism of the histone methyltransferase Pr-Set7

Methylation of lysine residues of histones is an important epigenetic mark that correlates with functionally distinct regions of chromatin. We present here the crystal structure of a ternary complex of the enzyme Pr-Set7 (also known as Set8) that methylates Lys 20 of histone H4 (H4-K20). We show that the enzyme is exclusively a mono-methylase and is therefore responsible for a signaling role quite distinct from that established by other enzymes that target this histone residue. We provide evidence from NMR for the C-flanking domains of SET proteins becoming ordered upon addition of AdoMet cofactor and develop a model for the catalytic cycle of these enzymes. The crystal structure reveals the basis of the specificity of the enzyme for H4-K20 because a histidine residue within the substrate, close to the target lysine, is required for completion of the active site. We also show how a highly variable component of the SET domain is responsible for many of the enzymes' interactions with its target histone peptide and probably also how this part of the structure ensures that Pr-Set7 is nucleosome specific.