A combination of isolated phytochemicals and botanical extracts lowers diastolic blood pressure in a randomized controlled trial of hypertensive subjects.

BACKGROUND/OBJECTIVES: Isolated phytochemicals have been shown to reduce blood pressure; however, combinations of phytochemicals have rarely been tested in humans. We hypothesized that a combination of extracts from grape seed and skin (330 mg), green tea (100 mg), resveratrol (60 mg) and a blend of quercetin, ginkgo biloba and bilberry (60 mg) would reduce blood pressure (BP) in hypertensive subjects. SUBJECTS/METHODS: Eighteen individuals meeting BP requirements (⩾130 mm Hg systolic or ⩾85 mm Hg diastolic) and criteria for metabolic syndrome were enrolled in a double-blinded, placebo-controlled, crossover trial (ClinicalTrials.gov, NCT01106170). The 28-day placebo and supplement arms were separated by a 2-week washout period, and 14 -h daytime ambulatory BP was assessed at baseline and at the end point of each arm. RESULTS: BP was not altered after placebo. After supplement treatment, diastolic pressure was reduced by 4.4 mm Hg (P=0.024, 95% CI, 0.6-8.1), systolic pressure was unchanged and mean arterial pressure trended (P=0.052) toward reduction. Serum angiotensin-converting enzyme activity was similar between placebo and supplement arms, but urinary nitrate and nitrite concentrations were significantly increased (P=0.022) after supplementation. Human aortic endothelial cells treated with metabolites of the polyphenols used in the human supplement trial had a significant increase (P=0.005) in insulin-stimulated eNOS phosphorylation and greater (P<0.001) accumulation of nitrates/nitrites. CONCLUSIONS: Our clinical and in vitro data support the theory that this combination of polyphenols reduced diastolic pressure by potentiating eNOS activation and nitric oxide production. Such supplements may have clinical relevance as stand-alone or adjunct therapy to help reduce BP.

Stereotactic radiotherapy for treatment of juxtapapillary choroidal melanoma: 3-year follow-up.

AIMS: To report the treatment outcomes and complication rates of stereotactic radiotherapy in the management of patients with juxtapapillary choroidal melanoma. METHODS: A retrospective review of 64 consecutive patients with juxtapapillary choroidal melanoma, located within 2 mm of the optic disc, treated with stereotactic radiotherapy at Princess Margaret Hospital between October 1998 and January 2006. RESULTS: The median age was 63 years. The median tumour height was 4.2 mm, and median largest basal diameter was 9.8 mm. The prescribed radiation dose was 70 Gy in five fractions over 10 days, and the median follow-up was 37 months. Post-treatment, the actuarial rates of local tumour control, metastases and survival at 37 months were 94%, 15% and 90%, respectively. Actuarial rates of radiation-induced complications at 37 months were neovascular glaucoma 42%, cataract 53%, retinopathy 81% and optic neuropathy 64%. Secondary enucleation was necessary for 10 patients (16%), in four patients for tumour recurrence and in six for painful neovascular glaucoma. CONCLUSIONS: Stereotactic radiotherapy offers a non-invasive alternative to enucleation and brachytherapy in the management of juxtapapillary choroidal melanoma with a high tumour control rate, however, at the expense of a significant rate of long-term ocular complications.

Effects of population size on performance and inbreeding depression in Lupinus perennis.

We investigated the relationships among population size, offspring performance, and inbreeding depression (delta) in Lupinus perennis by examining the effect of population size category (large vs. small) on seed production and offspring performance for three pollination treatments (open pollination, hand crossing and hand selfing). In each of our four pairs of populations, one member of the pair was substantially larger than the other. We then grew seeds from this factorial design (2 sizes x 4 pairs x 3 pollination treatments) in the greenhouse to investigate whether population size affects offspring performance in a common environment, and how small size affects purging of the inbreeding load. Multiplicative performance across four early life-stage components (seed production, seedling emergence, seedling survival and seedling growth) of smaller populations was not significantly lower, although biomass of seedlings declined in smaller populations. Self-pollination reduced seed production, seedling emergence and seedling growth, reflecting substantial inbreeding depression (delta = 0.404 +/- 0.043). Population size categories did not consistently differ in levels of inbreeding depression, suggesting that purging of genetic load in smaller populations has been limited, and that all populations still harbor inbreeding load. We also found a significant decrease in log performance with increases in the population inbreeding coefficient. These results indicate that even in seemingly large populations, lupines are susceptible to considerable fitness declines through both inbreeding load within populations, and drift load via genetic erosion and fixation of deleterious alleles between populations.

Focal stereotactic external beam radiotherapy as a vision-sparing method for the treatment of peripapillary and perimacular retinoblastoma: preliminary results.

AIMS: Chemotherapy with aggressive focal ablative therapy is now the mainstay of retinoblastoma therapy. Our experience presents an evolution from conventional radiotherapy by treating posterior pole tumours with focal stereotactic fractionated radiotherapy (SRT). MATERIALS AND METHODS: A retrospective chart review was conducted of five patients (six eyes) treated with SRT at the Hospital for Sick Children and Princess Margaret Hospital, Toronto, Canada, between 1999 and 2004. The prescribed dose was 40 Gy delivered in 20 fractions once daily using 6 MV photons. RESULTS: Five patients (six eyes) were treated. The median age at the time of SRT was 18 months. The median follow-up was 46.5 months as of September 2004. Four patients were treated for a posterior pole focal tumour by focal SRT, and one patient was treated for vitreous seeding with whole-eye SRT. In patients treated with focal SRT, the median doses to the tumour, optic chiasm and brainstem were 41.92, 0.25 and 0.07 Gy, respectively, and to the ipsilateral optic nerve, globe and lens were 9.98, 19.11 and 3.74 Gy, respectively. The median doses to the ipsilateral and contralateral orbital bone were 6.73 Gy (range 5.99-8.29 Gy) and 2.31 Gy (range 0.88-7.08 Gy), respectively. A complete response (residual inactive scar tissue) was seen in four of the five focal tumours treated, with one tumour responding with a partial response (suspicious residual scar tissue). No acute or late side-effects occurred in patients treated with focal SRT. Only the patient treated with whole-eye SRT developed late effects of cataract and corneal ulceration. One patient suffered recurrence within the radiation field 5 months after focal SRT. Control of this recurrence was successful using chemotherapy and focal therapy. No eye has been enucleated. CONCLUSION: Vision-sparing focal SRT for localised tumour masses in critical locations can control tumours with minimal side-effects and a minimal dose to the surrounding critical normal tissue.

Intraperitoneal therapy of malignant ascites associated with carcinoma of ovary and breast using radioiodinated monoclonal antibody 2G3.

A phase I/II study of intraperitoneal (ip) radioimmunotherapy was conducted in ovarian or breast cancer patients with symptomatic chemotherapy-resistant ascites using a novel anti-mucin monoclonal antibody (mAb) 2G3 labeled with 131I. Tracer doses of 2 mCi [131I]2G3 were given by ip injection to 11 patients, followed by increasing therapeutic doses up to 150 mCi (cumulative) in 9 patients. There was no serious toxicity. Temporary palliation of ascites was observed in 3 of 4 patients who received doses greater than 50 mCi. Total body elimination half-life of the radiolabeled antibody assessed by gamma scintigraphy ranged from 95 to 250 hr, longer than data previously reported in patients without ascites treated with ip administered radiolabeled antibodies. However, uptake of radiolabel by tumor nodules was small and variable (2 x 10(-4) - 2 x 10(-2) % ID/g), and preferential uptake by tumor compared to normal peritoneum was observed in only 2 of 5 patients in whom biopsies were obtained. These results suggest that the observed palliation of ascites is due to prolonged retention of radiolabeled antibody in the peritoneal cavity even in the absence of specific targeting.

Measurements in high-intensity beams from medical linear accelerators.

There is a potential to produce very high doses and dose rates on dual mode (electron and x-ray capability) medical electron accelerators [C. J. Karzmark, Br. J. Radiol. 40, 697 (1967)]. Measurements were taken on two accelerators to determine the size of the doses which were possible. The experiments demonstrate the limitations of many of the commonly used dosimeters when they are used outside of the dose and dose rate ranges normally encountered in a radiotherapy department. Dose rates from 110 to 165 cGy/pulse were measured using ionization chamber, ferrous sulphate and thermoluminescent dosimetry, and water calorimetry.

Oxygen depletion in irradiated aqueous solutions containing electron affinic hypoxic cell radiosensitizers.

The oxygen concentration in stirred aqueous solutions contained in sealed glass vessels was continuously monitored during irradiation with a sensitive Clark-type oxygen probe. The yield of radiolytic oxygen depletion, g(-O2), in alpha medium was determined to be about 0.44 microM/Gy (equivalent to 3.6 ppm/rad) over a range of oxygen from about 1,000 to 209,000 ppm. Over this same range of oxygen concentration, it was observed that oxygen is depleted in the presence of misonidazole, and that g(-O2) is slightly reduced at low oxygen and at high misonidazole concentrations. Oxygen depletion was observed in solutions of other nitroaromatic sensitizers of widely varying electron affinities: metronidazole, paranitroacetophenone, nitrofurazone, and nifurpipone. Significant protection of oxygen from radiolytic depletion was observed in concentrated solutions of nifurpipone, the most electron affinic drug studied (E17 = -214 mV). No such effect was observed for the least electron affinic compound, metronidazole (E17 = -486 mV).

Characteristics of electron beams from a new 25-MeV linear accelerator.

Clinically useful electron fields are produced on the Atomic Energy of Canada, Limited Therac 25 linear accelerator by computer-controlled scanning of the electron beam. Measurements were made to determine the properties of these electron fields. Central axis percentage depth dose and bremsstrahlung background were compared for these fields and for the fields from selected machines that use scattering foils. Dose calibrations were made in both water and polystyrene using the American Association of Physicists in Medicine Task Group 21 protocol. Measurements were made to determine the relative output factors, virtual source position, and the attenuation of the electron fields by lead.

Characteristics of the photon beam from a new 25-MV linear accelerator.

The Therac 25 is a relatively compact therapy machine, the heart of which is a double-pass electron linear accelerator. The electron beam is injected into the accelerator at the treatment head end of the machine and is accelerated back down the arm to an energy of 13 MeV. At this end of the machine a magnet system reflects the beam back into the structure where it gains up to an additional 12 MeV of energy. After leaving the linear accelerator the beam is bent by an achromatic head magnet through 270 degrees to the treatment head. The machine produces eight electron beams and a 25-MV photon beam. In this work only the parameters of the photon beam are addressed based on measurements at the first two clinical sites. Percentage depth doses, tissue phantom ratios, and beam symmetry and stability are presented and discussed.

Radiation protection aspects of a new high-energy linear accelerator.

The Therac-25 is a new 25-MeV linear accelerator manufactured by Atomic Energy of Canada, Ltd. The first two units have recently been installed in Toronto, Ontario and Halifax, Nova Scotia. Calculations and measurements of primary and secondary radiation levels were made. Neutron dose-equivalent rates were measured inside and outside the room. The maximum leakage rate at 1 m from the accelerator target was 0.4% Sv per peak photon Gy. The tenth value layer for neutrons from the Therac-25, at the entrance to a one-legged maze was found to be 5.5 cm of polyethylene. Measurements were done to estimate daily technologist exposure due to induced activity in the treatment room.

The oxygen dependence of the reduction of nitroimidazoles in a radiolytic model system.

Radiation chemical reductions using eaq- and CO2- have been carried out in the presence of oxygen with metronidazole, p-nitroacetophenone, misonidazole and three other 2-nitroimidazoles. Low concentrations of oxygen were found to effectively inhibit the reduction of the first two compounds while much higher concentrations of oxygen were required for all of the 2-nitroimidazoles. These results parallel in vitro and in vivo experiments with metronidazole and misonidazole which also indicate that the reduction of the latter is significantly less inhibited by oxygen. Kinetic modelling of the radiochemical system suggests that the explanation for the differences lies in different reactions of the nitro radical anions; it appears that the anion derived from metronidazole undergoes disproportionation while that derived from misonidazole undergoes a unimolecular decay.

Increased radiosensitivity of aerobic mammalian cells following pre-incubation with misonidazole in hypoxia.

The effect of pre-incubation of CHO cells with misonidazole in hypoxia on the aerobic radiosensitivity of these same cells was observed and analyzed according to the concepts of independent and additive interaction. It was found that the sensitivity of cells irradiated in air, in the presence or absence of misonidazole, increased when the cells had been pre-incubated with misonidazole in hypoxia at 37 degrees C. This increase in aerobic radiosensitivity was evident as a loss of the shoulder of the survival curve, and the data were consistent with an additive, rather than an independent interaction mechanism. The effect was also observed when the cells were washed prior to irradiation. These data are consistent with our previous work with hypoxic cells, and support the hypothesis that a toxic metabolite of the sensitizer is produced during hypoxic incubation, whose effect is manifested as an additional increment of cell killing equivalent to an additional dose of radiation. Given the possibility of a diffusible toxic product of sensitizer nitroreduction, the hypoxic cell toxicity of nitroimidazoles, generally perceived to be of importance only in hypoxic cells, may be a factor in the radiosensitivity of non-hypoxic cells in tumours.

Night shift work.

The deleterious effects of night shift work are discussed in relation to emergency physicians. Much of the early research is drawn from heavy industry sources and must be extrapolated to medical work. Disruption of circadian rhythms and Zeitgebers, caused by adherence to contemporary social schedules, causes recurrent difficulty for physicians who constantly change schedules. Adjustments in both physical surroundings and social structure that can make the effects of traumatic schedule change more tolerable are discussed.

Effects of modifiers of the yield of hydroxyl radicals on the radiosensitivity of mammalian cells at ultrahigh dose rates.

Experiments were conducted to investigate the effects of increasing or decreasing the yield of hydroxyl radicals (.OH) reacting with target molecules on the survival of CHO cells irradiated in a thin layer with single 3-nsec pulses of electrons. Dimethyl sulfoxide (DMSO), a known radioprotector, was used as an .OH scavenger. The gas nitrous oxide (N2O), which scavenges hydrated electrons and in the process generates an additional yield of .OH, was used in an attempt to produce sensitization by increasing the amount of .OH-induced cellular damage. It was found that DMSO at high concentration was an effective radioprotective agent in cells equilibrated with nitrogen, air, and N2O and irradiated at ultrahigh dose rates. Sensitization by N2O was observed, but only under certain conditions, specifically, when a high concentration of the .OH scavenger DMSO was also present. The enhancement ratio (ER) for oxygen sensitization was reduced in the presence of DMSO, as was the ER for sensitization by misonidazole. Interpretation of these results according to radiation chemistry considerations will be discussed.

Irradiation of mammalian cells in the presence of diamide and low concentrations of oxygen at conventional and at ultrahigh dose rates.

The response of cultured CHO cells to ultrahigh-dose-rate radiation (approximately 10(9) Gy/sec) has been previously studied extensively using the thin-layer cell-handling technique developed in this laboratory. When the cells are equilibrated with a low concentration of oxygen, e.g., 0.44% O2, a breaking survival curve, due to radiolytic depletion of the oxygen, is observed. Hypoxic cells irradiated in the presence of the nitroimidazoles (e.g., misonidazole) are sensitized at ultrahigh dose rates in a dose-modifying manner, similar to that observed at conventional dose rates. These radiosensitizer compounds, if present in cells equilibrated with a low concentration of oxygen, prevent the breaking behavior of the survival curve, an observation believed to be due to the sensitizer interfering with the oxygen depletion process, leaving oxygen free to sensitize. Such experiments have recently been extended to studies with diamide, which, unlike the other sensitizers tested, acts primarily as a shoulder-modifying rather than a dose-modifying agent in hypoxic mammalian cells. These data indicate that diamide is active as a sensitizer at ultrahigh dose rates in a manner similar to that observed at conventional dose rates, and does modify the shape of the breaking survival curve observed with low concentrations of oxygen.

Influence of low pH on the development and decay of 42 degrees C thermotolerance in CHO cells.

The development, magnitude and decay of thermotolerance in Chinese hamster ovary cells heated to 42 degrees C at pH 6.7 was examined. The cells were exposed to single or fractionated heat treatments with 0 to 168 hours elapsing between the treatments. Administration of a specified heat treatment in two fractions at low pH substantially reduced the lethal effect of heat. The rate of hyperthermic cell killing was reduced by a factor of approximately 2.5 in preheated cells compared to cells not receiving prior heat treatment. Tolerance to second heat treatments was apparent when 3 hours at 37 degrees C separated the treatments, and was near maximum when 6 hours separated the treatments. Beginning about 96 hours after the initial heat treatment, resistance to second heat treatments began to decline, and was not evident when 168 hours at 37 degrees C separated the treatments. In contrast to these results, a sparing effect as a result of dose fraction was not observed at pH 7.4. At pH 7.4, cells developed thermotolerance during the initial heat treatment and additional culturing at 37 degrees C was without additional effect. Nevertheless, the sensitivity of cells to hyperthermia was greater at pH 6.7 than at pH 7.4. This pH sensitizing effect was more pronounced in cells exposed to single heat treatments than in cells exposed to fractionated treatments at 42 degrees C.

Effect of electron affinic hypoxic cell sensitizers on the radiolytic depletion of oxygen in mammalian cells irradiated at ultrahigh dose rates.

When CHO cells are equilibrated with a low level of oxygen (e.g. 0.4% O2) and irradiated with single 3 ns pulses of electrons, a breaking survival curve is observed. This effect is believed to be the result of radiolytic oxygen depletion and can be prevented by the presence of a relatively low concentration of hypoxic cell radiosensitizer. This prevention of the breaking survival curve has been observed for 2- and 5-nitroimidazoles, nitrofurans, and diamide. It is hypothesized that the sensitizer acts by competing with oxygen for the radiation-induced intracellular oxygen-binding species, perhaps a hydrated electron adduct, leaving oxygen free to participate in radiosensitization reactions during the lifetime of the oxygen-sensitive radiation-induced target sites for lethal damage, probably DNA radicals produced by hydroxyl radical attack. The proposed role of the sensitizer in the interference with oxygen depletion is a transient phenomenon, occurring on the microsecond to millisecond time scale.