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PubMed1995

Island hopping.

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Family Practice↗
PubMed1997

-GG- specific cleavage via hole hopping through double-stranded DNA.

We examined the photoinduced DNA cleavage reaction of 32P-end labeled oligodeoxyribonucleotides which contain 5'-TGGGC-3', 5'-CGGGC-3', and 5'-TGGGT-3'. As for 5'-TGGGC-3' and 5'-TGGGT-3', the cleavage occurred most strongly at the middle of 5'-GGG-3' step, whereas the strongest cleavage occurred at the 5' side of 5'-GGG-3' in 5'-CGGGC-3' step. These results showed the 5' side base of 5'-GGG-3' step was very important for the sequence selectivity of 5'-GGG-3' cleavage.

Base Sequence↗
PubMed2000

UVA-Induced DNA single-strand cleavage by 1-hydroxypyrene and formation of covalent adducts between DNA and 1-hydroxypyrene.

1-Hydroxypyrene (HOP), a metabolite found in the urine of humans and laboratory animals exposed to polycyclic aromatic hydrocarbons (PAHs), is known to be both acutely toxic and genotoxic. It has been widely used as a biomarker for studying PAH exposure. In this research, we have found that, upon UVA irradiation, HOP causes DNA single-strand cleavages and forms HOP-DNA covalent adducts. The UVA-induced cleavage of supercoiled plasmid PhiX174 DNA is dependent upon both HOP concentration and UVA dosage. A longer irradiation time or higher HOP concentration induces more DNA cleavage. Results of the photocleavage experiments carried out in the presence of reactive oxygen species scavengers, histidine, sodium azide, mannitol, SOD, and desferal indicate that both the superoxide free radical and singlet oxygen are likely involved in causing DNA single-strand cleavage. The photocleavage is inhibited by the presence of an excited singlet-state quencher, KI, indicating that it is an excited-state reaction. Along with light-induced DNA cleavage, HOP also forms DNA covalent adducts while being degraded upon light irradiation. Light-induced degradation of 20 microM HOP follows first-order reaction kinetics in a 10% methanolic buffer (10 mM phosphate) solution in the absence or presence of 40 microM calf thymus DNA, with degradation half-lives of 20 or 15 min, respectively. The shorter degradation half-life in the presence of DNA is due to the formation of the HOP-DNA covalent adduct. The formation of the HOP-DNA covalent adduct is evidenced by comparing the UV-vis absorption and fluorescence emission spectra of the pure HOP with those of the HOP-DNA adduct. The covalent HOP-DNA adduct produced due to irradiation was purified by either extensive dialysis (3 x 500 mL buffer solutions), phenol and chloroform extraction followed by ethanol precipitation, or chloroform extraction alone. The isolated HOP-DNA adduct has an absorption peak at 353 nm, which is 8 nm red-shifted compared to that of free HOP. The fluorescence emission for HOP-DNA is at least 70 times weaker than that for free HOP in solution. In summary, the findings with HOP reveal that, in addition to metabolic activation that eventually leads to the formation of alkylated DNA adducts or other forms of DNA damage, HOP may be activated by light to produce DNA single-strand cleavage and covalent DNA adducts. These DNA lesions can be sources of toxicity.

Animals↗
PubMed2000

Long-term evaluation of bone formation by osteogenic protein 1 in the baboon and relative efficacy of bone-derived bone morphogenetic proteins delivered by irradiated xenogeneic collagenous matrices.

To investigate the long-term efficacy of irradiated recombinant human osteogenic protein 1 (hOP-1) in bone regeneration and morphogenesis, hOP-1 was combined with a bovine collagenous matrix carrier (0, 0.1, 0.5, and 2.5 mg hOP-1/g of matrix), sterilized with 2.5 Mrads of y-irradiation, and implanted in 80 calvarial defects in 20 adult baboons (Papio ursinus). The relative efficacy of partially purified bone-derived baboon bone morphogenetic proteins (BMPs), known to contain several osteogenic proteins, was compared with the recombinant hOP-1 device in an additional four baboons. Histology and histomorphometry on serial undecalcified sections prepared from the specimens harvested on day 90 and day 365 showed that gamma-irradiated hOP-1 devices induced regeneration of the calvarial defects by day 90, although with reduced bone area compared with a previous published series of calvarial defects treated with nonirradiated hOP-1 devices. One year after application of the irradiated hOP-1 devices, bone and osteoid volumes and generated bone tissue areas were comparable with nonirradiated hOP-1 specimens. Moreover, 365 days after healing regenerates induced by 0.5 mg and 2.5 mg of irradiated hOP-1 devices showed greater amounts of bone and osteoid volumes when compared with those induced by nonirradiated hOP-1 devices. On day 90, defects treated with 0.1 mg and 0.5 mg of bone-derived baboon BMPs, combined with irradiated matrix, showed significantly less bone compared with defects receiving irradiated devices containing 0.1 mg and 0.5 mg hOP-1; 2.5 mg of partially purified BMPs induced bone and osteoid volumes comparable with the 0.1-mg and 0.5-mg hOP-1 devices. Control specimens of y-irradiated collagenous matrix without hOP-1 displayed a nearly 2-fold reduction in osteoconductive bone repair when compared with nonirradiated controls. These findings suggest that the reduction in bone volume and bone tissue area on day 90 may be caused by a reduced performance of the irradiated collagenous matrix substratum rather than to a reduction in the biological activity of the irradiated recombinant osteogenic protein. This is supported by the results of in vitro and in vivo studies performed to determine the structural integrity of the recovered gamma-irradiated hOP-1 before application in the baboon. Recoveries by high-performance liquid chromatography (HPLC) and sodium dodecyl sulfate/ polyacrylamide gel electrophoresis (SDS/PAGE)/immunoblot analyses indicated that doses of 2.5-3 Mrads of gamma-irradiation did not significantly affect the structural integrity of the recovered hOP-1. Biological activity of the recovered hOP-1 was confirmed in vitro by showing induction of alkaline phosphatase activity in rat osteosarcoma cells (ROS) and in vivo by de novo endochondral bone formation in the subcutaneous space of the rat. These findings in the adult primate indicate that a single application of gamma-irradiated hOP-1 combined with the irradiated xenogeneic bovine collagenous matrix carrier is effective in regenerating and maintaining the architecture of the induced bone at doses of 0.5 mg/g and 2.5 mg/g of carrier matrix.

Alkaline Phosphatase↗
PubMedDate not supplied

Pharmacokinetics and metabolism of the pharmacologically active 4'-hydroxylated metabolite of propranolol in the dog.

The disposition of 4'-hydroxypropranolol (HOP) was determined after iv administration to dogs (2 mg/kg; N = 5) and the pharmacokinetic parameters were calculated from plasma measurements. The clearance of HOP, 66 +/- 6 ml/min/kg (mean +/- SE), was considerably higher than that of propranolol previously determined, suggesting extrahepatic as well as hepatic clearance of HOP. The plasma half-life of HOP, 77 +/- 6 min, was shorter than that of propranolol. Although HOP is considerably less lipophilic than propranolol, its volume of distribution, 6.4 +/- 0.8 liter/kg, surprisingly, was larger. Like propranolol, HOP appeared to be cleared entirely by metabolism. Whereas propranolol is metabolized mainly by oxidation, HOP was metabolized to sulfate (HOPS) and glucuronic acid (HOPG) conjugates. The plasma half-lives of these conjugates were 2 to 3 times longer than for HOP, reflecting a slow, continuous formation from HOP. This was established for HOPS by iv administration of synthetic HOPS. Morover, after HOP administration both formation and renal clearance of HOPS were stereoselective in favor of the R-enantiomer. In summary, the main conclusion of this study is that the large volume of distribution as well as high clearance through sulfation and glucuronidation may explain the low plasma HOP levels observed during propranolol therapy.

Animals↗
PubMed1997

Prevention of heterotopic ossification about the hip: final results of two randomized trials in 410 patients using either preoperative or postoperative radiation therapy.

PURPOSE: Experimental and clinical data support effectiveness of perioperative radiotherapy to prevent heterotopic ossification after hip surgery or trauma. Since 1987, two prospectively randomized trials were performed in patients with high-risk factors to develop heterotopic ossification: the first (HOP 1) to assess the prophylactic efficacy of postoperative low vs. medium dose radiotherapy, and the second (HOP 2) to assess the prophylactic efficacy of pre vs. postoperative radiotherapy. METHODS AND MATERIAL: 410 patients with high risk to develop heterotopic ossifications about the hip following hip surgery were recruited. Between June 1987 and June 1992, 249 patients were randomized in HOP 1 to postoperative "low dose" (5 x 2 Gy; total: 10 Gy) or "medium dose" (5 x 3.5 Gy; total: 17.5 Gy) radiotherapy. Between July 1992 and December 1995, 161 patients were randomized in HOP 2 to either 1 x 7 Gy preoperatively (< or = 4 h before surgery) or 5 x 3.5 Gy (total: 17.5 Gy) postoperatively (< or = 96 h after surgery). With exception of age and type of implant (cemented vs. uncemented prosthesis) all confounding patient variables (gender, prior surgery) and predisposing risk factors were similarly distributed between both trials and treatment arms. Portals encompassed the periacetabular and intertrochanteric soft tissues. Radiographs were obtained prior and immediately after surgery and at least 6 months after surgery to assess the extent of ectopic bone formation about the hip. Modified Brooker grading was used to score the extent of heterotopic ossification. Harris scoring was applied to evaluate the functional hip status. If the scores decreased from immediate post or preoperative status, respectively, to the last follow-up, radiological or functional failures were assumed. RESULTS: Effective prophylaxis was achieved in 227 (91%) hips of HOP 1 and in 142 (88%) of HOP 2. In HOP 1, 15 (11%) radiological failures were observed in the low-dose group compared to 7 (6%) in the medium dose group (p > 0.05). In HOP 2, 4 (5%) radiological failures were observed in the postoperative and 11 (19%) in the preoperative group (p < 0.05). Subgroup analysis of the preoperative group revealed that the highest failure rate occurred in patients with prophylactic radiotherapy prior to removal of ipsilateral Brooker Grade III and IV ossification (39%) (p < 0.001), while all other patients in the preoperative group had a failure rate that was comparable to postoperative treatment groups. In multivariate logistic regression analysis the number of high-risk factors for development of heterotopic ossification (p = 0.03) and the time to RT initiation (p = 0.05) were independent prognostic factors in the HOP 1 study. For the HOP 2 study, the multivariate logistic regression analysis revealed the number of high-risk factors for development of heterotopic ossification (p = 0.003), the preoperative HO grade (p = 0.001) and the RT dose concept (p = 0.05) as independent prognostic factors. Other factors including type of implant (cemented vs. uncemented) did not affect the prophylactic efficacy of radiotherapy. There were no increased intra- and postoperative complications seen in the preoperative group, and no long-term complications were observed in both HOP studies. For functional failures (decrease of Harris score) no statistically prognostic factors were found. There were less functional failures in HOP 1 (18 = 7%) than in HOP 2 (23 = 14%, but this difference was not statistically significant. Only patients with high Brooker Grade III and IV at last FU achieved a lower Harris score than those with low Brooker Grade 0, I and II (p < 0.05). CONCLUSION: With the exception of a small subgroup of patients with ipsilateral high Brooker Grade III and IV, pre- and postoperative radiotherapy are equally effective to prevent heterotopic ossification about the hip after hip surgery and total hip arthroplasty. Fractionated medium dose radiotherapy resulted in the low

Adult↗
PubMed1997

Mutation in the Jak kinase JH2 domain hyperactivates Drosophila and mammalian Jak-Stat pathways.

The Jak (Janus) family of nonreceptor tyrosine kinases plays a critical role in cytokine signal transduction pathways. In Drosophila melanogaster, the dominant hop(Tum-l) mutation in the Hop Jak kinase causes leukemia-like and other developmental defects. Previous studies have suggested that the Hop(Tum-l) protein might be a hyperactive kinase. Here, we report on the new dominant mutation hop(T42), which causes abnormalities that are similar to but more extreme than those caused by hop(Tum-l). We determined that Hop(T42) contains a glutamic acid-to-lysine substitution at amino acid residue 695 (E695K). This residue occurs in the JH2 (kinase-like) domain and is conserved among all Jak family members. We determined that Hop(Tum-1) and Hop(T42) both hyperphosphorylated and hyperactivated D-Stat when overexpressed in Drosophila cells. Moreover, we found that the hop(T42) phenotype was partially rescued by a reduction of wild-type D-stat activity. Finally, generation of the corresponding E695K mutation in murine Jak2 resulted in increased autophosphorylation and increased activation of Stat5 in COS cells. These results demonstrate that the mutant Hop proteins do indeed have increased tyrosine kinase activity, that the mutations hyperactivate the Hop-D-Stat pathway, and that Drosophila is a relevant system for the functional dissection of mammalian Jak-Stat pathways. Finally, we propose a model for the role of the Hop-D-Stat pathway in Drosophila hematopoiesis.

Amino Acid Sequence↗
PubMed2005

Functional-Performance Deficits in Volunteers With Functional Ankle Instability.

Context: Although functional-performance tests are dynamic measures used to assess general lower body function, studies investigating these tests for ankle instability have yielded conflicting results.Objective: To determine if a relationship exists between a measure of functional ankle instability and deficits in functional performance.Design: A case-control study correlating subject performance on a set of lower extremity functional-performance tests with a measure of ankle instability.Setting: University athletic training research laboratory.Patients or Other Participants: We recruited 60 participants (43 females, 17 males, age = 22.4 +/- 4.9 years, height = 169.9 +/- 9.7 cm, mass = 72.6 +/- 16.3 kg; 42 injured, 18 uninjured) to participate in the study. Six questions were used to determine if functional ankle instability was present in each participant. A point was added for each yes response to produce an index that represents a continuous variable of functional ankle instability.Main Outcome Measure(s): Four unilateral hopping tests were used in this study: figure-of-8 hop, side hop, up-down hop, and single hop. For the first 3 tests, the total time was recorded with a handheld stopwatch to the nearest 0.01 second; for the single hop-for-distance test, the distance was recorded to the nearest 0.01 m. Correlations were computed with the functional ankle instability index and each of the 4 functional-performance tests.Results: No relationship was revealed between the functional ankle instability index and single hop for distance or up-down hop, with r values of -.008 and .245, respectively. A significant relationship was found between the functional ankle instability index and the side hop (r = .35, P </= .01) and the figure-of-8 hop (r = .31, P </= .02).Conclusions: A positive relationship existed between functional ankle instability and performance deficits on the side hop and figure-of-8 hop. Conversely, a relationship did not exist between functional ankle instability and frontal-plane functional-performance activities.

Journal Article↗
PubMed2006

Selective inhibition of COX-2 by a standardized CO2 extract of Humulus lupulus in vitro and its activity in a mouse model of zymosan-induced arthritis.

A standardized CO(2) extract from Humulus lupulus L. (hop extract) was investigated for its selective COX-1/2 inhibitory properties. An in vitro model of inflammation using lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMC) was used as a model to investigate the effect of hop extract on PGE(2) production. COX-1/2 selective inhibition by the hop extract was investigated in a COX-1 whole blood assay (WBA) and a COX-2 WBA. To evaluate the in vivo activity of hop extract, it was administered orally to C57BL/6 mice in which inflammation of the right joint was induced by injecting zymosan intra-articularly. Ex vivo PGE(2) production of LPS-stimulated blood cells was determined. Also, the effect of hop extract on healthy and arthritic cartilage was investigated as well as effects on inflammatory joint swelling. Hop extract inhibited PGE(2) production by LPS-stimulated PBMC without compromising the metabolic activity of these cells. Furthermore, hop extract showed a decline in PGE(2) production in the COX-2 whole blood assay (WBA) with an IC(50) of 20.4 microg/mL, while in the COX-1 WBA no inhibition of PGE(2) production was observed. This indicates a COX-2 selective inhibition. The COX-1 inhibitor SC-560 inhibited PGE(2) production in the COX-1 WBA but not in the COX-2 WBA. At 2 microM, celecoxib inhibited PGE(2) production in the COX-2 WBA by 92 % and in the COX-1 WBA by 50 %. When hop extract was administered orally to C57BL/6 mice in which joint inflammation was induced with zymosan, PGE(2) production in ex vivo LPS-stimulated whole blood was significantly decreased by 24 %, suggesting that hop extract becomes bioavailable. Furthermore, oral administration of hop extract showed no negative or positive effects on healthy cartilage proteoglycan synthesis, or on zymosan-induced arthritic cartilage proteoglycan synthesis. However, no effect of oral administration of 1.25 mg hop extract daily was observed on joint swelling. In conclusion, this standardized CO(2) extract of Humulus lupulus could be a useful agent for intervention strategies targeting inflammatory disorders and/or inflammatory pain.

Administration, Oral↗
PubMed1988

The aerobic cost of saltatory locomotion in the fowler's toad (Bufo woodhousei fowleri).

Studies of kangaroos suggest that hopping provides energy savings during locomotion at high speeds, although studies of small mammals suggest that hopping is no more economical than running. To obtain comparative data on anurans, we exercised Fowler's toads (Bufo woodhousei fowleri, 25.8 g) on treadmills at speeds ranging from 0.09 to 0.63 km h-1 while measuring oxygen consumption (VO2), endurance or hop kinematics. The toads walked at slow speeds and hopped at fast speeds. Steady-state VO2 (VO2,ss) increased linearly with speed to a maximum (VO2, max) of 1.17 ml O2 g-1 h-1 at 0.27 km h-1 and was nine times the average pre-exercise VO2. The maximum rate of oxygen consumption during treadmill exercise was comparable to VO2,max previously reported for less natural exercise regimes. At speeds greater than or equal to 0.27 km h-1, VO2,ss was independent of speed. At speeds less than or equal to 0.36 km h-1, toads moved for over 1h, but endurance decreased sharply at higher speeds. Hop rate, hop length, hop height and angle of take-off increased with speed. Hopping in B.w. fowleri was not less costly than running in other animals of similar body size and was inefficient at converting metabolic to mechanical energy. The present study suggests that hopping in toads, as in small mammals, is not economical during sustained locomotion and is most important during short bursts of high-intensity activity.

Animals↗
PubMed1997

Reliability of lower extremity functional performance tests.

Clinicians routinely have used functional performance tests as an evaluation tool in deciding when an athlete can safely return to unrestricted sporting activities. These practitioners assumed that these tests provide a reliable measure of lower extremity performance; however, little research has been reported on the reliability of these measures. The purpose of this investigation was to determine the reliability of lower extremity functional performance tests. Five male and 15 female volunteers were evaluated using the single hop for distance, triple hop for distance, 6-m timed hop, and cross-over hop for distance as described by Noyes (10). One clinician measured each subject's performance using a standardized protocol and retested subjects in the same manner approximately 48 hours later. The order of testing was randomly determined. Subjects' average and individual scores on each functional performance test were used for statistical analysis. Intraclass correlation coefficients (ICCs) and standard error of measurement (SEM) values based on average day 1 and day 2 scores were used to estimate the reliability of each functional performance test. Intraclass correlation coefficients were .96, .95, and .96, and SEMs were 4.56 cm, 15.44 cm, and 15.95 cm, respectively, for the single hop, triple hop, and cross-over hop for distance tests. An ICC of .66 and SEM of .13 seconds for the 6-m timed hop resulted from limited variability between measurements; however, its small SEM value inferred that the inconsistency of measurement would occur in an acceptably small range. A repeated measures analysis of variance revealed no significant difference ( p > .05) between individual trial scores except for the single hop for distance. We concluded that this difference represented a learning effect not found with the other tests. The results of this investigation demonstrate that clinicians can use functional performance testing to obtain reliable measures of lower extremity performance when using a standardized protocol.

Adult↗
PubMed1985

A review of urinary hydroxyproline as a biochemical marker on health effects of smoking and air pollution with nitrogen dioxide.

A series of epidemiologic studies on the health effects caused by smoking and air pollution with NO2 were carried out. Urinary hydroxyproline (HOP) which is excreted into the urine by catabolism of lung collagen was used as a biochemical marker. Urinary HOP of smokers increased dose-dependently with increasing numbers of cigarettes smoked and this result was accepted under blind conditions. Urinary HOP of nonsmokers also increased with increasing the amounts of involuntary smoking caused by smokers in their families although the increases in HOP by passive smoking were less than those by active smoking. Health effects caused by indoor air pollution with NO2 in winter and automobile exhaust in summer were also demonstrated using urinary HOP. The effects of cessation of smoking were followed up for 14 weeks by the decrease in urinary HOP. In practice, the HOP to creatinine ratio (HOP ratio) in random urine samples was used as representative of the 24-hour urine samples. Some confounding factors for urinary HOP were taken into consideration in these studies.

Air Pollution↗