A novel simple phantom for verifying the dose of radiation therapy.

A standard protocol of dosimetric measurements is used by the organizations responsible for verifying that the doses delivered in radiation-therapy institutions are within authorized limits. This study evaluated a self-designed simple auditing phantom for use in verifying the dose of radiation therapy; the phantom design, dose audit system, and clinical tests are described. Thermoluminescent dosimeters (TLDs) were used as postal dosimeters, and mailable phantoms were produced for use in postal audits. Correction factors are important for converting TLD readout values from phantoms into the absorbed dose in water. The phantom scatter correction factor was used to quantify the difference in the scattered dose between a solid water phantom and homemade phantoms; its value ranged from 1.084 to 1.031. The energy-dependence correction factor was used to compare the TLD readout of the unit dose irradiated by audit beam energies with (60)Co in the solid water phantom; its value was 0.99 to 1.01. The setup-condition factor was used to correct for differences in dose-output calibration conditions. Clinical tests of the device calibrating the dose output revealed that the dose deviation was within 3%. Therefore, our homemade phantoms and dosimetric system can be applied for accurately verifying the doses applied in radiation-therapy institutions.

Laboratory and field evaluation of measurement methods for one-hour exposures to O3, PM2.5, and CO.

While researchers have linked acute (less than 12-hr) ambient O3, PM2.5, and CO concentrations to a variety of adverse health effects, few studies have characterized short-term exposures to these air pollutants, in part due to the lack of sensitive, accurate, and precise sampling technologies. In this paper, we present results from the laboratory and field evaluation of several new (or modified) samplers used in the "roll-around" system (RAS), which was developed to measure 1-hr O3, PM2.5, and CO exposures simultaneously. All the field evaluation data were collected during two sampling seasons: the summer of 1998 and the winter of 1999. To measure 1-hr O3 exposures, a new active O3 sampler was developed that uses two nitrite-coated filters to measure O3 concentrations. Laboratory chamber tests found that the active O3 sampler performed extremely well, with a collection efficiency of 0.96 that did not vary with temperature or relative humidity (RH). In field collocation comparisons with a reference UV photometric monitor, the active O3 sampler had an effective collection efficiency ranging between 0.92 and 0.96 and a precision for 1-hr measurements ranging between 4 and 6 parts per billion (ppb). The limits of detection (LOD) of this method were 9 ppb-hr for the chamber tests and approximtely 16 ppb-hr for the field comparison tests. PM2.5 and CO concentrations were measured using modified continuous monitors--the DustTrak and the Langan, respectively. A size-selective inlet and a Nafion dryer were placed upstream of the DustTrak inlet to remove particles with aerodynamic diameters greater than 2.5 microm and to dry particles prior to the measurements, respectively. During the field validation tests, the DustTrak consistently reported higher PM2.5 concentrations than those obtained by the collocated 12-hr PM2.5 PEM samples, by approximately a factor of 2. After the DustTrak response was corrected (correction factor of 2.07 in the summer and 2.02 in the winter), measurements obtained using these methods agreed well with R2 values of 0.87 in the summer and 0.81 in the winter. The results showed that the DustTrak can be used along with integrated measurements to measure the temporal and spatial variation in PM2.5 exposures. Finally, during the field validation tests, CO concentrations measured using the Langan were strongly correlated with those obtained using the reference method when the CO levels were above the LOD of the instrument [approximately 1 part per million (ppm)].

Paracrine function of cultured aortic endothelial cells in spontaneously hypertensive rats.

BACKGROUND: Vasoconstrictor and vasodilator release from vascular endothelial cells not only regulates vascular tone but also induces vascular smooth muscle cell proliferation. METHODS: In order to understand the role of vasoconstrictor and vasodilator release in the development of hypertension in spontaneously hypertensive rats (SHR), aortic endothelial cells were isolated and cultured from 4-week-old and 24-week-old SHR (SHR-4 and SHR-24) and age-matched Wistar-Kyoto rats (WKY-4 and WKY-24) used as control. Prostacyclin (PGI2), endothelin-1 (ET-1) and thromboxane A2 (TXA2) release from cultured endothelial cells in the culture medium, were measured after 30 min with or without treatment with acetylcholine, calcium ionophore A23187 or thrombin. RESULTS: The results showed that there was no significant difference in ET-1 secretion between SHR-4 and age-matched WKY rats, but ET-1 secretion was about twice as high in SHR-24 as in WKY-24. TXA2 secretion was significantly higher in SHR-4 than in WKY-4 and was also higher than in SHR-24, but there was no significant difference between SHR-24 and WKY-24. The secretion of PGI2 was higher in SHR-24 than in WKY-24 and also higher than in SHR-4 and WKY-4. The prostaglandin PGI2 and TXB2 secretions from all groups of cultured VECs treated with various reagents, acetylcholine, calcium ionophore A23187 or thrombin were increased in similar patterns. However, there was no significantly different response between SHR and WKY VECs. CONCLUSIONS: Similar levels of ET-1 secreted from endothelial cells between SHR-4 and WKY-4 indicated that ET-1 secretion seems not a crucial factor in early hypertension development in SHR. The high level of TXA2 secretion in SHR-4 may involve in early hypertension development in SHR.

Temporal and spatial sequence expression of cytokeratin K19 in cultured human keratinocyte.

The unique cytokeratin K19 specifically expresses in simple epithelial cells, basal cells of non-keratinized stratified squamous epithelium, epidermal cells during the embryonic stage and squamous carcinoma cells, but it is not expressed in adult epidermis. Interestingly, when epidermal cells are cultured in vitro, K19 is re-expressed in the supra-basal layer. K19 expression was used as a marker for epidermal cell growth and differentiation. In order to clarify the temporal and spatial sequential expression in cultured keratinocyte, two-stage human keratinocyte culture systems were used to examine K19 expression in keratinocytes in a proliferation and differentiation stages through immunoblotting and immunohistochemistry assay. According to our results, K19 was not expressed in cultured human keratinocytes in the proliferation stage but was re-expressed in keratinocytes three days after the cultured medium was changed to a differentiation medium. Immunohistochemical observation revealed that K19 was persistently expressed in the supra-basal layer of cultured keratinocytes during first three weeks of culturing, but none was detectable in the basal cell layer. When keratinocytes were cultured with an "inserted cultured dish," K19 was persistently expressed in all layers of keratinocytes nourished by medium both from an inner chamber and an outer chamber. The different expression of K19 in these two different culture systems seemed to indicate that down regulation of K19 expression in keratinocyte was related to the direction of medium supply.

Hourly personal exposures to fine particles and gaseous pollutants--results from Baltimore, Maryland.

A study to characterize 1-hr multi-pollutant exposures was performed in Baltimore, MD, during the summer of 1998 and the winter of 1999, and was conducted over a 15-day period in each of the two seasons. Personal exposures were measured by a trained field technician, who wore a newly developed Roll-Around System (RAS) to measure 1-hr PM2.5 and gaseous (CO, O3, NO2, SO2, volatile organic compounds [VOCs]) exposures. One-hour O3, NO2, and SO2 personal exposures were measured using samplers developed in our laboratory, while short-term PM2.5, CO, and VOCs exposures were measured using currently available monitors. All 1-hr multi-pollutant exposures were measured while the technician performed pre-determined activities, beginning at 7:00 a.m. and ending at 7:00 p.m. of the same day. Activities were scripted to simulate activities performed by older adults (65+ years of age). Corresponding 1-hr ambient pollutant concentrations were obtained from federal or state monitoring networks. In this paper, we discuss the results from our study and present our descriptive analysis of the 1-hr personal particulate and gaseous exposure data. Personal PM2.5, O3, CO, and VOCs exposures showed substantial variability over the 12-hr sampling periods. Multiple pairwise comparison tests showed that 1-hr personal O3 exposures were significantly lower in indoor microenvironments as compared with outdoor microenvironments. One-hour personal CO exposures measured in vehicles were significantly higher than those measured in other microenvironments. The associations between 1-hr personal exposures and corresponding ambient concentrations differed by pollutant and by microenvironment. For example, the correlation between personal PM2.5 exposures and ambient concentrations was lowest (rs = 0.36, p < 0.05) in the winter for indoor non-residential microenvironments, and was highest (rs = 0.90, p < 0.05) in the winter for in-vehicle microenvironments. For O3, the correlation between personal exposures and ambient levels was weakest in the winter for residential microenvironments (rs = 0.05, p > 0.05), and was strongest in the summer for outdoor near-roadway microenvironments (rs = 0.91, p < 0.05).

Head injury in facial impact--a finite element analysis of helmet chin bar performance.

The chin bar of a motorcycle helmet protects the rider from facial and head injuries. To evaluate the protective performance of chin bars against head injuries from facial impacts, an explicit finite element method was used to simulate the Snell Memorial Foundation test and a proposed drop test. The maximum acceleration and Head Injury Criterion (HIC) were employed to assess the impact-absorbing capability of the chin bar. The results showed that the proposed approach should be more practical than the Snell test, and provided more information for improving the chin bar design to protect against head injuries. The shell stiffness was important in determining the protective ability of the chin bar, but a chin bar with only an outer shell and comfort foam offered inadequate protection. An energy-absorbing liner was essential to increase the protective performance of the chin bar and the liner density should be denser than that used in the cranial portion of the helmet. For the chin bar with energy-absorbing liner, a shell design that is less stiff would provide better protection.

Gonadal transformation in male Rana catesbeiana tadpoles intraperitoneally implanted with estradiol capsules.

Tadpoles of Rana catesbeiana at TK stages X-XII, about 9 months old, were laparotomized, and the males were implanted intraperitoneally with silastic tubes containing estradiol (E2) for various periods. Male tadpoles implanted with empty tubes served as the controls. Histology, secretions of E2, and testosterone (T) in the gonads were investigated. A rough estimate of estradiol released from silastic tubes suggested that about 90 micrograms per tadpole in 6 months. Histological observation revealed various degrees of transformation from testes toward ovaries in E2-treated testes. Ten in thirteen (77%), the testes were transformed into ovaries 6 months after the treatment. The testes of the controls, however, displayed normal histology. Radioimmunoassay showed that E2 level was increased while T level was decreased in E2-treated testes. These results indicate that a low dose of exogenous E2 may induce transformation of the testes into ovaries.

Craniofacial injuries in unhelmeted riders of motorbikes.

We studied 1160 consecutive craniofacial injuries sustained by unhelmeted motorbike riders in Taipei, Taiwan, between 1990 and 1993, in order to investigate the distribution, type and severity of these injuries. The average age of the victims was 31 years (SD 13.2), with 84 per cent of them being between ages 16 and 45. The facial and cranial areas were defined as being separated by the line between eyebrows and ears. The incidence of facial injuries was the same as that of cranial injuries (both 68 per cent). While facial injuries occurred most often in the cheek and chin, most cranial injuries occurred in the forehead and parietal region. Although the majority of facial injuries resulted in mild brain injuries, they may also cause serious cosmetic problems, and some were associated with serious brain damage. Motorbike riders need good face protection. Since most motorbikes in Taipei travel relatively slowly, these results may also apply to bicyclists; in other words, cyclists may also need good face protection.

Voluntary running in male S5B/P1Ras rats fed high fat or high carbohydrate diets.

Nineteen week-old male S5B/P1Ras (S5B) rats were randomly assigned to one of 4 groups as follows: (a) activity wheel access (running)/high fat diet (RF); (b) no activity wheel access (non-running)/high fat diet (NRF); (c) activity wheel access (running)/high carbohydrate diet (RC); and (d) no activity wheel access (non-running)/high carbohydrate diet (NRC) for the seven weeks duration of the experiment. Throughout the 7 wk of the experiment, rats ran more during subsequent weeks than they did the previous week. RC rats ran more than RF rats as measured by the running slopes. All groups of rats lost weight at the initiation of the experiment but significantly more weight was lost by running rats than their nonrunning counterparts. The inguinal, epididymal and perirenal/retroperitoneal (P/R) fat depots weighed significantly less in the running than in the nonrunning groups. From among the 3 fat depots, the difference was greatest in the P/R depot. There were no diet or voluntary activity effects on plasma corticosterone concentrations except at week 2 when running rats had higher concentrations than nonrunning rats.

Gonadal differentiation and secretions of estradiol and testosterone of the ovaries of Rana catesbeiana tadpoles treated with 4-hydroxyandrostenedione.

Laparotomized female tadpoles of Rana catesbeiana at TK stages X-XII, about 9 months old, were implanted intraperitoneally with empty capsules or capsules containing 4-hydroxyandrostenedione (4-OHA), known as an aromatase inhibitor in vertebrates. Histology, gonosomatic index, and secretions of estradiol (E2) and testosterone (T) of the ovaries were investigated. Three months after the treatment, histological examination revealed various degrees of sex reversal in the ovaries treated with 4-OHA and 79% (57 in 72) were transformed into testes. The ovaries of control tadpoles, however, displayed normal histological appearance. Radioimmunoassay showed that secretion of E2 was decreased while that of T was increased in 4-OHA treated ovaries. The gonosomatic index displayed a decline tendency from control females through experimental animals to untreated control males. These results indicated that activity of aromatase in the ovaries was inhibited by 4-OHA, resulting in accumulation of T which induced transformation of the ovaries into testes.

Decreased production of para-hydroxypenicillin V in penicillin V fermentations.

Penicillin V (phenoxymethyl penicillin) is produced by industrial strains of Penicillium chrysogenum in the presence of phenoxyacetic acid (POAc), a side-chain precursor for the penicillin V molecule. The wild-type strain of P. chrysogenum produces an undesirable penicillin byproduct, para-hydroxypenicillin V (p-OH penicillin V), in addition to penicillin V, via para-hydroxylation of POAc and subsequent incorporation of the p-OH phenoxyacetic acid into the penicillin molecule. Most of the p-OH penicillin V is produced late in cycle when the POAc concentration in the medium is nearly depleted. The level of p-OH penicillin V produced by the control strain ranges up to 10-15% of the total penicillins produced. 3-Phenoxypropionic acid and p-bromophenylacetic acid partially inhibit the formation of p-OH penicillin V with a minimal effect on penicillin V productivity. Mutants deficient in their ability to hydroxylate POAc were found to produce lower levels of p-OH penicillin V. Multi-step mutation and screening, starting with the wild-type strain, have culminated in isolation of mutants which produce p-OH penicillin V as 1% of the total penicillins with no adverse effect on penicillin V productivity.

Penicillin production by glucose-derepressed mutants of Penicillium chrysogenum.

Wild-type strains of Penicillium chrysogenum produce lower penicillin V titers in media containing excess glucose. Two mutant strains were isolated and shown to produce normal penicillin V titers in the presence of excess glucose. These strains, designated as glucose-repression insensitive (GRI) mutants, produced higher penicillin V titers than the wild-type strain in media containing lactose as the main carbohydrate source. In lactose-based media, the production of penicillin V was depressed to a much lesser extent by in-cycle additions of glucose with the GRI mutants when compared to the wild-type strain. In short-term biosynthesis experiments using washed cells in a medium containing glucose as the sole carbon source, the GRI mutants produced penicillin V at a faster rate than the wild-type strain. In fed-batch fermentations in 14-liter fermentors, where glucose was fed continuously and pH controlled, both GRI mutants produced more than 10% higher penicillin V titers than the wild-type strain. These results suggest that isolation of GRI mutants is an effective way to select for higher producing strains and that the synthesis of penicillin synthesizing enzymes in GRI mutants may be less repressed by glucose than in wild-type strains.

Estradiol secretion by the ovarian tissue, in response to hypophyseal stimulation, during ontogenesis of the bullfrog.

Ovaries of tadpoles, froglets, young frogs, and mature frogs of Rana catesbeiana were cut into small pieces. They were incubated for 6 hr in Krebs-Ringer bicarbonate buffer as controls. Another series of ovaries of the same developmental stages were incubated with pituitary extracts in the buffer as experimentals. Media were then analyzed for estradiol secretion by radioimmunoassay. The results showed that estradiol secretion by tadpole ovaries during development was not affected by the addition of pituitary extracts of mature frogs in the media at any stage while those of young and mature frogs with pituitary extracts secreted more estradiol than those without. These findings indicate that tadpole ovaries are unresponsive to pituitary agents to produce estradiol while frog ovaries are dependent on some pituitary hormones to synthesize estradiol. Thus frog ovaries acquire dependence on the pituitary agent only after metamorphosis.

Estrogen suppression of preincubated tadpole ovaries in vitro by cyanoketone.

In a previous RIA study we found that cyanoketone (CK) inhibited ovarian E2 secretion of tadpoles in vitro and that this inhibition effect was through inactivation of delta 5-3 beta-HSD activity. A complete 100% inhibition was expected at a CK dosage of 0.1 microgram/ml of the medium, but, instead, it was 85%. The discrepancy might be due to the fact that the previous experiments did not preincubate the ovaries with CK in order to get rid of the residual E2. To this end, the present study was designed. Tadpole ovaries of Rana catesbeiana were preincubated with CK of a dosage of 0, 0.001, 0.01, 0.1, 1, or 10 micrograms/ml of the KRbb medium for 30 min. The media were discarded. Fresh media with the same series of CK doses were added to the ovaries and were incubated again for 6 hr. The media were collected for RIA of estrogen. The results showed the same tendency of estrogen inhibition as the previous study. However, a maximal inhibition effect of 95% was obtained at the dose of 0.1 microgram/ml. Therefore, the difference between non-preincubation of the previous experiments and preincubation of the present study does exist as we predicted.

In vitro inhibition of estradiol secretion of tadpole ovaries by cyanoketone.

We have demonstrated the presence of delta 5-3 beta-hydroxysteroid dehydrogenase (HSD) activity in tadpole ovaries of Rana catesbeiana. In the present study, we wish to determine whether estradiol (E2) secretion of tadpole ovaries could be influenced by cyanoketone (CK), a specific inhibitor of delta 5-3 beta-HSD. R catesbeina tadpoles at the premetamorphic climax were used, and pooled ovaries were incubated, 30 mg/tube, with CK at dosages of 0, 0.001, 0.01, 0.1, 1, and 10 micrograms/ml of Krebs-Ringer bicarbonate buffer for 6 hr. Media were collected for assay of E2 by radioimmunoassay (RIA). Results showed an inhibition of E2 secretion by CK that was positively correlated with CK dosage, but plateaued at doses of 0.1 microgram/ml and higher. This finding was comparable to that of G.F. Young, H. Kagawa, and Y. Nagahama (1982, Gen. Comp. Endocrinol. 47, 357-360) on adult fish ovaries. However, adult vertebrates depend on gonadotropins to regulate secretion of E2 while tadpoles, being immature, might secrete E2 independently of pituitaries. When the histochemical test for delta 5-3 beta-HSD activity was performed on in vitro CK-treated ovaries, there was a decrease of enzyme activity by CK. The RIA and histochemical findings may contribute to the concept of sex transformation in which a disturbance of steroidogenesis may induce sex reversal from females to males, at least in tadpoles.

Contrasting effects of ACTH and cyanoketone on delta 5-3 beta-hydroxysteroid dehydrogenase activity in interrenal glands of tadpoles of Rana catesbeiana in vitro.

The present communication describes an investigation of stimulation and inhibition of delta 5-3 beta-hydroxysteroid dehydrogenase in interrenal glands of tadpoles of Rana catesbeiana. Frozen sections of interrenal glands, together with kidneys, were prepared histochemically for assay of delta 5-3 beta-HSD activity. Concentrations of 0.01, 0.1, 1, and 10 IU/ml of ACTH or of 0.01, 0.1, 1, and 10 micrograms/ml of cyanoketone were added to the incubation media. The reaction products of the histochemically prepared slides, in terms of absorbance, were scanned at a defined area with a computerized microscope spectrophotometer. The results indicate that ACTH causes a significant dose-response stimulation of delta 5-3 beta-HSD activity in tadpole interrenals; cyanoketone, on the other hand, causes significant dose-dependent inhibition.

In vitro estradiol synthesis and secretion by tadpole ovaries of different developmental stages.

The tadpole ovaries at TK stage XIX can synthesize and secrete estradiol (E2), yet it is unknown when this ovarian function starts and how it develops. To this end, the present work has been carried out. The ovaries of different developmental stages of tadpoles and young frogs of Rana catesbeiana were taken, cut into small pieces, and incubated for 6 hr at 20 degrees in Krebs-Ringer bicarbonate buffer. The media were then analyzed for E2 by radioimmunoassay. The results indicate that the tadpoles synthesize and release E2 as early as stage X, that E2 increases slowly and gradually through stage XXV, and that E2 increases rapidly in juvenile frogs. This trend of growth of the ovarian function in estradiol secretion in vitro is in accordance with that reported for female chick embryos. There exists the possibility that the growth of E2 secretion could be biphasic, one with slow increment of autonomous E2 secretion at early stages and the other with quick increase due to pituitary stimulation at the stage of metamorphic climax.

Molecular basis of an inherited retinal defect in Drosophila.

Fruitflies carrying the autosomal recessive mutation transient receptor potential (trp) are blind in bright light because the receptor potential of such a mutant decays almost completely during an intense stimulus. The trp gene has been localized and a set of partially overlapping genomic clones that include the trp gene has been isolated. The stretch of DNA represented by these genomic clones is found to contain genes that encode for four RNA species. Two of these RNA species are missing in the mutant. This observation is consistent with the notion that the mutation alters the DNA sequence in a region containing signals necessary for the expression of the gene. Accordingly, the molecular basis of the mutant phenotype may be due to the lack of a protein(s) that is/are important for normal visual transduction.

Positively charged filters for virus recovery from wastewater treatment plant effluents.

Positively charged Zeta Plus filters were used to concentrate enteroviruses from 19 liters of effluent from activated sludge units. Neither the addition of salts nor the acidification of the effluent was required for adsorption of viruses to the filters. Viruses adsorbed to the filters were eluted by treating the filters with a solution of 4 M urea buffered at pH 9 with 0.05 M lysine. Eluted viruses were concentrated into final volumes of 1 to 2 ml by using a two-step concentration procedure that employed inorganic and organic flocculation. Approximately 50% of the viruses added to effluents could be recovered in the final sample. The procedure was used to monitor effluents from activated sludge units at two wastewater treatment plants for the presence of enteroviruses.

Recovery of coliphages from wastewater effluents and polluted lake water by the magnetite-organic flocculation method.

A magnetite-organic flocculation method was developed for the concentration of coliphages from wastewater effluents and polluted lake water. A high percent (68 to 100%) recovery of coliphages from sewage effluents was achieved by this procedure. Coliphage recovery from Lake Alice, a sewage-contaminated lake, showed phage concentrations ranging from 2.3 X 10(2) to 1.9 X 10(3) plaque-forming units per liter. This method is simple and inexpensive and may be carried out under field conditions.