Barriers to patient recruitment in a poststroke neurorehabilitation multicenter trial in Brazil.

There is a high demand for stroke rehabilitation in the Brazilian public health system, but most studies that have addressed rehabilitation for unilateral spatial neglect (USN) after stroke have been performed in high-income countries. Therefore, the aim of this study was to analyze USN patient recruitment in a multicenter noninvasive brain stimulation clinical trial performed in Brazil and to provide study design recommendations for future studies. We evaluated the reasons for exclusion of patients from a multicenter, randomized, double-blinded clinical trial of rehabilitation of USN patients after stroke. Clinical and demographic variables were compared between the included and excluded patients. A descriptive statistical analysis was performed. Only 173 of the 1953 potential neglect patients (8.8%) passed the initial screening. After screening evaluation, 87/173 patients (50.3%) were excluded for clinical reasons. Cognitive impairment led to the exclusion of 21/87 patients (24.1%). Low socioeconomic status led to the exclusion of 37/173 patients (21.4%). Difficulty obtaining transportation to access treatment was the most common reason for their exclusion (16/37 patients, 43.3%). The analyzed Brazilian institutions have potential for conducting studies of USN. The recruitment of stroke survivors with USN was restricted by the study design and limited financial support. A history of cognitive impairment, intracranial stenting or craniectomy, and lack of transportation were the most common barriers to participating in a multicenter noninvasive brain stimulation trial among patients with USN after stroke.

Barriers to patient recruitment in a poststroke neurorehabilitation multicenter trial in Brazil

There is a high demand for stroke rehabilitation in the Brazilian public health system, but most studies that have addressed rehabilitation for unilateral spatial neglect (USN) after stroke have been performed in high-income countries. Therefore, the aim of this study was to analyze USN patient recruitment in a multicenter noninvasive brain stimulation clinical trial performed in Brazil and to provide study design recommendations for future studies. We evaluated the reasons for exclusion of patients from a multicenter, randomized, double-blinded clinical trial of rehabilitation of USN patients after stroke. Clinical and demographic variables were compared between the included and excluded patients. A descriptive statistical analysis was performed. Only 173 of the 1953 potential neglect patients (8.8%) passed the initial screening. After screening evaluation, 87/173 patients (50.3%) were excluded for clinical reasons. Cognitive impairment led to the exclusion of 21/87 patients (24.1%). Low socioeconomic status led to the exclusion of 37/173 patients (21.4%). Difficulty obtaining transportation to access treatment was the most common reason for their exclusion (16/37 patients, 43.3%). The analyzed Brazilian institutions have potential for conducting studies of USN. The recruitment of stroke survivors with USN was restricted by the study design and limited financial support. A history of cognitive impairment, intracranial stenting or craniectomy, and lack of transportation were the most common barriers to participating in a multicenter noninvasive brain stimulation trial among patients with USN after stroke.

AN INTERNATIONAL COOPERATION BY USING AN ALL-ENCOMPASSING PASSIVE RADON MONITOR.

The recently developed radon film-badge makes it possible to measure radon indoors, in soil, in water and/or in aqueous media (e.g. mud). As a result of its wide response linearity, this monitor has been successfully used to measure radon in-water with concentrations from 10 to ~10 000 Bq/L. By exploiting the unique characteristics of this badge, a mini-survey has been carried out by Health Canada in which radon in water was measured from 12 private wells, as well as in tap water originating from the Ottawa River. Due to the widespread interest of different laboratories in using these passive monitors, laboratories were provided with plastic films to construct their own badges by using in-house CR-39 detectors. Monitors were then irradiated by a known radon concentration at the National Institute of Radiation Metrology (ENEA)'s radon chamber and sent back to each laboratory for processing and counting. Even though these laboratories have been using different etching- and counting-procedures, the film-badge responses varied only within ~12%.

Exploring the effects of immunity and life history on the dynamics of an endogenous retrovirus.

Mammalian DNA is littered with the signatures of past retroviral infections. For example, at least 8% of the human genome can be attributed to endogenous retroviruses (ERVs). We take a single-locus approach to develop a simple susceptible-infected-recovered model to investigate the circumstances under which a disease-causing retrovirus can become incorporated into the host genome and spread through the host population if it were to confer an immunological advantage. In the absence of any fitness benefit provided by the long terminal repeat (LTR), we conclude that signatures of ERVs are likely to go to fixation within a population when the probability of evolving cellular/humoral immunity to a related exogenous version of the virus is extremely small. We extend this model to examine whether changing the speed of the host life history influences the likelihood that an exogenous retrovirus will incorporate and spread to fixation. Our results reveal the parameter space under which incorporation of exogenous retroviruses into a host genome may be beneficial to the host. In our final model, we find that the likelihood of an LTR reaching fixation in a host population is not strongly affected by host life history.

Modified repeated intracyclic clomiphene citrate therapy after conventional clomiphene therapy.

PURPOSE OF INVESTIGATION: We compared modified repeated intracyclic clomiphene citrate therapy (RICCT) to gonadotropin therapy to determine whether this modified regimen was an effective alternative after conventional clomiphene therapy. METHODS: Patients with ovulation disorder received treatment with modified RICCT and gonadotropin, and ovulation, pregnancy, total drug cost, and adverse effects were compared. RESULTS: Among a total of 16 patients, 14 successfully ovulated after modified RICCT and 11 ovulated after gonadotropin therapy; two did not respond to either therapy. The total drug cost was US $36.3+/-17.9 for modified RICCT, which was significantly lower than the cost of gonadotropin therapy, US $213.9+/-100.4 (p=0.0001). CONCLUSIONS: Because modified RICCT does not require the discomfort of daily injection and has excellent ovulation-inducing effects, it is a useful treatment after conventional clomiphene therapy.

Chromosome aberrations in peripheral blood lymphocytes of individuals living in high background radiation areas of Ramsar, Iran.

In order to investigate the biological effects of exposure to low-dose radiation and to assess the dose-effect relationship in residents of high background radiation areas (HBRAs) of Ramsar, cytogenetic investigation of unstable-type aberrations was performed in 15 healthy elderly women in a HBRA of Ramsar, Talesh mahalle, and in 10 elderly women living in a nearby control area with normal background radiation. In total, 77,714 cells were analyzed; 48,819 cells in HBRA residents and 28,895 cells in controls. On average, 3,108 cells per subject were analyzed (range 1,475-5,007 cells). Significant differences were found in the frequency of dicentric plus centric rings in 100 cells (0.207 ± 0.103 vs. 0.047 ± 0.027, p < 0.0005), total chromosome-type aberrations per 100 cells (0.86 ± 0.44 vs. 0.23 ± 0.17, p < 0.0005), and chromatid-type aberrations per 100 cells (3.31 ± 2.01 vs. 1.66 ± 0.63, p = 0.01) by the Mann-Whitney U test between HBRA and the control, respectively. Using chromosomal aberrations as the main endpoint to assess the dose-effect relationship in residents of HBRAs in Ramsar, no positive correlation was found between the frequency of dicentric plus centric ring aberrations and the cumulative dose of the inhabitants estimated by direct individual dosimetry; however, obvious trends of increase with age appeared in the control group. Based on these results, individuals residing in HBRAs of Ramsar have an increased frequency of detectable abnormalities in unstable aberrations.

Population diversity in batches of the varicella Oka vaccine.

The Oka vaccine is a live attenuated vaccine for the prevention of varicella. Although the vaccine differs from the progenitor virus by over 40 mutations, only three of these are fixed, the rest being a mixture of the wildtype and the vaccine allele. To examine the extent of this variability between two of the three commercially available vaccine preparations, we analysed the vaccine/wildtype allele frequencies present at fifteen vaccine loci in five preparations each from two different manufacturers of the vOka vaccine. Our results suggest that differences in manufacturing processes between the two companies have resulted in significant variation in the frequencies of the vaccine/wildtype alleles in their vaccines. Yet despite these differences, the allele frequencies in the vaccines from the two companies are strongly correlated. We discuss the significance of these findings and the role of evolutionary processes that influence the production of this live attenuated vaccine.

Repeated intracyclic clomiphene citrate therapy can be more effective than hMG therapy in inducing ovulation: case report.

PURPOSE OF INVESTIGATION: When clomiphene citrate is ineffective in the treatment of anovulation, hMG administration is typically selected. However, high-dose hMG therapy is associated with a variety of adverse events. We describe the use of a modified clomiphene citrate regimen that was successful in increasing the effectiveness of ovulation induction. CASE REPORT: A patient who did not initially respond to clomiphene citrate therapy required a total dose of 2400 IU hMG to prodeuce mature follicles. However, because of the physical and emotional burdens on the patient, and the possibility of multiple pregnancy and ovarian hyperstimulation syndrome, re-treatment with clomiphene citrate was then selected. Two courses of clomiphene citrate administered at a fixed interval during the same cycle safely induced ovulation. After initial induction of ovulation, her ovulatory failure improved and natural ovulation occurred. CONCLUSIONS: Repeated intracycle clomiphene cirate therapy may be more effective than hMG therapy in inducing ovulation in some patients.

Removal of endocrine disrupting compounds during conventional wastewater treatment.

There is evidence that aquatic organisms downstream of some sewage treatment works show endocrine disruption as a result of exposure to substances in the effluent. As a result, the Environment Agency of England and Wales, in collaboration with the UK Government and the water industry, has started an intensive programme to determine the fate and behaviour of endocrine disrupting compounds in sewage treatment works. Sampling sites for the endocrine disruption demonstration programme are located throughout England and Wales. This paper presents data from Nuneaton sewage treatment works (Warwickshire, England), a modem nitrifying activated sludge plant serving an equivalent population of 98,000 and one of the selected sites for the demonstration programme. Results for the 24-hour survey carried out in June 2006 in which manual grab samples were taken hourly show excellent removal of estrone, estradiol, nonyl-phenol and the nonylphenolethoxylates (3-5 EO units) at 97, 99, 94 and 98% respectively. They also show excellent removal (99%) of estrogenicity, measured by the YES bioassay. However the removal of ethynylestradiol was poor at only 3%. In November 2006, a further survey was carried out comprising grab samples taken at 4-hourly intervals across a continuous 7-day period. This monitoring confirmed the good removal of estrone and estradiol, at 97.8% and 96.3% respectively as well as an excellent reduction in estrogenicity (98.3%), but again showed poor removal of ethynylestradiol of 5.6%. There was evidence of a diurnal pattern for estrone and estradiol concentrations and to a lesser extent for ethynylestradiol in samples of crude sewage with works returns. Peak concentrations tended to occur at around midday.

Chemically induced premature chromosome condensation in short-term cultured human peripheral lymphocytes: applications to biodosimetry.

We describe here the chemical induction of premature condensed chromosomes in human peripheral lymphocytes after culture for 6 h. Many have attempted this induction without culture or with short-term culture, because this technique permits prompt cytogenetic biodosimetry of radiation accidents. Lymphocytes were separated from blood, incubated in the presence of phytohemagglutinin, ATP, and p34(cdc2)/cyclin B kinase, then treated with calyculin A during the last hour. The culture medium was supplemented with a lower concentration of fetal calf serum than conventionally used to minimize its possible interference with the effects of these drugs. We obtained, rarely, a suitable morphology of premature chromosome condensation in short-term cultured lymphocytes for conventional chromosome aberration analysis.

Investigation of new cytogenetic biomarkers specific to high-LET radiation using in vivo and in vitro exposed human lymphocytes.

PURPOSE: To find detectable cytogenetic biomarkers that can offer information about the radiation quality of in vivo exposure retrospectively. MATERIALS AND METHODS: Chromosome-type aberrations of peripheral lymphocytes of uterine cancer patients that received internal gamma- and external X-ray therapy or carbon beam therapy and of victims severely exposed to neutrons and gamma-rays in a criticality accident that occurred in Tokai-mura, Japan were analysed. Data obtained from in vitro irradiation experiments using 60Co gamma-rays and 10 MeV neutrons were compared with the in vivo exposure data. RESULTS: The ratio of acentric rings to dicentric chromosomes (termed RaD ratio) and that of excess fragments to dicentrics (termed EfD ratio) showed significant (p < 0.05) differences between the two groups of cancer patients, and these ratios for accidental victims were in between the values of the two groups of cancer patients. The in vitro studies using doses equivalent to 1 - 3 Gy of gamma-rays have confirmed that the EfD ratios were increased with the high LET (linear energy transfer) and RaD ratios decreased. CONCLUSION: The present data show that the RaD and EfD ratios can be used as cytogenetic biomarkers of exposure to high-LET radiation at least within a few years of exposure.

Sister chromatid exchanges in ring chromosomes following X-irradiation of human lymphocytes.

PURPOSE: To examine whether X-rays induce sister chromatid exchanges (SCE). MATERIALS AND METHODS: Peripheral lymphocytes irradiated in vitro or in vivo were cultured and treated with okadaic acid to generate premature chromosome condensation (PCC). When identical spreads were analysed using conventional Giemsa staining and pan-centromeric fluorescence in situ hybridization painting, ring chromosomes were observed. RESULTS: In PCC preparations, cells in the late G(2) phase and late M phase were observed. In late M phase cells, 17-20% of ring chromosomes lacked one chromatid (single-chromatid ring), irrespective of dose. Both the distribution patterns of centromeres in rings and intercentromere distances in dicentric rings indicate that a considerable number of single-chromatid rings might be formed by SCE occurring in a chromosome-type ring, thereby joining strands of two rings, followed by a transformation into one ring. These single-chromatid rings were less stable in vivo than chromosome-type rings. CONCLUSION: Single-chromatid rings visualized clearly using PCC techniques indicate SCE in the respective rings. Contrary to the conventional SCE-detecting technique, this approach does not require the use of bromodeoxyuridine, which itself leads to SCE. Some of the observed SCE might be secondary products resulting from the repair of radiation-induced DNA damage, while others may be spontaneous.

Dynamic analysis of chromosome aberrations in three victims of the Tokai-mura criticality accident.

PURPOSE: To investigate the dynamics of chromosome aberrations in the blood cells of three workers severely exposed to neutrons and gamma-rays in a criticality accident that occurred in Tokai-mura, Japan, in 1999. MATERIALS AND METHODS: The change with time of the frequency of' chromosome aberrations in the three patients was examined using a new analysis to score drug-induced prematurely condensed ring chromosomes (PCC-R) and a conventional meta-phase analysis. RESULTS: The frequencies and cellular distributions of PCC-R, dicentrics and rings did not change significantly among the samples obtained at 9-48h after the accident while the first depletion of lymphocytes occurred. The distributions of these aberrations in the cells of two patients showed a slight overdispersion compared with a Poisson distribution reflecting neutron and non-uniform exposures. The dose response curve of rings paralleled that of dicentrics, but not PCC-R. The half-lives of PCC-R (8.5 months) and of rings (8.7 months) were shorter than that of dicentrics (13.5 months). CONCLUSIONS: In the three patients of the Tokai-mura accident, lymphocytes in the circulating and extravascular pools had reached equilibrium at 9h, and highly damaged lymphocytes did not selectively move away from the circulatory system during the first rapid depletion of lymphocytes after exposure. Data on the in vivo half-life of PCC-R as well as dicentrics and rings obtained in the present study may be useful for retrospective dosimetry.

Cytogenetical dose estimation for 3 severely exposed patients in the JCO criticality accident in Tokai-mura.

A dose estimation by chromosome analysis was performed on the 3 severely exposed patients in the Tokai-mura criticality accident. Drastically reduced lymphocyte counts suggested that the whole-body dose of radiation which they had been exposed to was unprecedentedly high. Because the number of lymphocytes in the white blood cells in two patients was very low, we could not culture and harvest cells by the conventional method. To collect the number of lymphocytes necessary for chromosome preparation, we processed blood samples by a modified method, called the high-yield chromosome preparation method. With this technique, we could culture and harvest cells, and then make air-dried chromosome slides. We applied a new dose-estimation method involving an artificially induced prematurely condensed ring chromosome, the PCC-ring method, to estimate an unusually high dose with a short time. The estimated doses by the PCC-ring method were in fairly good accordance with those by the conventional dicentric and ring chromosome (Dic+R) method. The biologically estimated dose was comparable with that estimated by a physical method. As far as we know, the estimated dose of the most severely exposed patient in the present study is the highest recorded among that chromosome analyses have been able to estimate in humans.

Improvement of accuracy of chromosome aberration analysis for biological radiation dosimetry.

The frequency of chromosome aberrations in circulating lymphocytes is accepted as being the most reliable indicator of the absorbed dose of radiation. Researches done to improve the accuracy of cytogenetic analysis are described in this review. These include investigations of in vitro factors that affect the yield of radiation-induced aberrations and of in vivo factors that affect the chromosomal radiosensitivity of individuals. Improved chromosome-painting methods for accurate judgment of dicentrics and translocations are introduced. The practicality of these advanced cytogenetic techniques is shown by examinations of individuals exposed in the radiation accident at Tokaimura in 1999.

Effect of estradiol on radiation-induced chromosome aberrations in human lymphocytes.

As a part of studies on physiological factors that affect radiosensitivity, we examined the in vitro effect of estradiol (E2) on the yield of radiation-induced chromosome aberrations in human peripheral lymphocytes. Lymphocytes were cultured for 3 days in the medium containing E2 at 0-100,000 ng/ml. On the second day, they were irradiated by X-rays at 3 Gy, and then 2% phytohemagglutinin and 0.05 microgram/ml colcemid were added to the medium. After further 48 h, mitotic indices and the yields of chromosome aberrations were examined at various E2 concentrations. E2 treatment at concentrations above 1000 ng/ml resulted in dose-related inhibition of mitosis. Repeated experiments showed that the yield of dicentrics plus centric rings in the culture containing E2 at 100 ng/ml was significantly higher than the yields at 0 ng/ml. Similarly, the yield of total chromosome breaks in the culture containing E2 at 100 ng/ml was significantly higher than that at 1 ng/ml. This study provides the direct evidence in human that radiosensitivity may vary in relation to hormonal conditions.

Easy biodosimetry for high-dose radiation exposures using drug-induced, prematurely condensed chromosomes.

PURPOSE: To propose a practical method of biodosimetry for high-dose estimation. MATERIALS AND METHODS: Okadaic acid in the range 100 nM to 500 nM was given for 1-6 h to cultured lymphocytes that had been exposed in the range of 0 Gy to 40 Gy. The frequency of the cells with prematurely condensed chromosomes (PCC) was compared with that of metaphases accumulated by 0.05 microg/ml colcemid. The dose-response relationship of the yield of prematurely condensed ring chromosomes (PCC rings) in the irradiated lymphocytes was examined. RESULTS: The 0.05 microg/ml colcemid collected few metaphases when lymphocytes were irradiated to over 10 Gy. However, 500 nM okadaic acid produced a substantial number of analysable PCC in lymphocytes that had been exposed in the range of 0 Gy to 40 Gy. After 1 h of treatment, the 500 nM okadaic acid gave the most suitable chromosome morphology for the analysis. PCC rings were narrow, which facilitated their detection in Giemsa-stained preparations. The yield of PCC rings increased dose-dependently up to 20 Gy. CONCLUSION: PCC rings are a more suitable indicator than dicentrics at high doses over 10 Gy. This technique requires no particular equipment or exceptional skill. After a serious radiation accident, it can be easily performed in a short time in most medical facilities to obtain dose information to assist physicians.

Allocyclic X chromosome visualized by drug-induced premature chromosome condensation.

Calyculin A, an inhibitor of serine/threonine protein phosphatases, can induce premature chromosome condensation (PCC) in human peripheral lymphocytes. In female lymphocytes treated with calyculin, one highly condensed chromosome per nucleus was frequently observed at early to middle S phase. When these PCC spreads were hybridized with a whole chromosome-painting probe for the X chromosome, one large spot and a cluster of small spots were observed in a cell. Judging from the Giemsa stained image, the former seemed to have finished DNA replication, while the latter was in the process of replication. Because this large spot was seldom observed in male PCC spreads, it was considered an inactive X chromosome. The inconsistency of this observation with earlier reports that the inactive X chromosome replicates later than autosomes and the active homolog is discussed.

Phosphatase inhibitors and premature chromosome condensation in human peripheral lymphocytes at different cell-cycle phases.

The cytogenetical reaction of human peripheral lymphocytes to okadaic acid and calyculin A was examined. Calyculin A could induce PCC about 20 times more effectively than okadaic acid. Their mechanisms of PCC induction were judged similar by their dose-dependent manner and chromosome morphology. Contrary to earlier studies suggesting that chemicals could not induce PCC in G1 cells where little cyclin B is present, the present study showed that calyculin A could induce PCC in lymphocytes not only at S and G2/M but also at the second G1 phase after PHA stimulation in vitro. PCC was induced slightly in lymphocytes both at G0 and the first in vitro G1 phase even when the calyculin A concentration increased one hundred fold. It was found that calcium ionophore A23187 increased frequencies of G0-PCC induced by calyculin A, although a further refinement is necessary to obtain a suitable morphology of G0-PCC for cytogenetic studies.

Chemically induced premature chromosome condensation in human fibroblast cell lines: fundamental study for applications to the biodosimetry of local exposure.

The premature chromosome condensation (PCC) of human peripheral lymphocytes treated with inhibitors of protein phosphatase has been demonstrated to be an excellent tool for the estimation of high-dose whole-body exposure. To develop a new biodosimetry for local exposure, the cytogenetical reaction of human fibroblast lines to PCC inducers was examined and compared with that of lymphocytes. The efficiency of the induction by calyculin A was greater than that by okadaic acid in both cell types. Calyculin A induced PCC in 5-Gy-irradiated and unirradiated samples at almost the same frequency in the lymphocytes, whereas the efficacy was considerably lower in irradiated fibroblasts than in unirradiated ones. Calcium ionophore enhanced the induction of PCC in irradiated fibroblasts, although PCC frequencies were still much lower than those in the lymphocytes. The frequency of ring chromosomes observed in 2- and 5-Gy-irradiated fibroblasts was too low to be used as a marker for cytogenetic dosimetry, and that of excess fragments, scored as the observed chromosome number minus 46, might be substituted. The frequency of excess fragments for 2-, 5-, and 10-Gy-irradiated fibroblasts was less than 0.75, about 1 and a few per cell, respectively, although these values changed with the culture period. The prospects and limitations of the application of PCC techniques to fibroblasts are discussed.