Syndromic surveillance system based on near real-time cattle mortality monitoring.

Early detection of an infectious disease incursion will minimize the impact of outbreaks in livestock. Syndromic surveillance based on the analysis of readily available data can enhance traditional surveillance systems and allow veterinary authorities to react in a timely manner. This study was based on monitoring the number of cattle carcasses sent for rendering in the veterinary unit of Talavera de la Reina (Spain). The aim was to develop a system to detect deviations from expected values which would signal unexpected health events. Historical weekly collected dead cattle (WCDC) time series stabilized by the Box-Cox transformation and adjusted by the minimum least squares method were used to build the univariate cycling regression model based on a Fourier transformation. Three different models, according to type of production system, were built to estimate the baseline expected number of WCDC. Two types of risk signals were generated: point risk signals when the observed value was greater than the upper 95% confidence interval of the expected baseline, and cumulative risk signals, generated by a modified cumulative sum algorithm, when the cumulative sums of reported deaths were above the cumulative sum of expected deaths. Data from 2011 were used to prospectively validate the model generating seven risk signals. None of them were correlated to infectious disease events but some coincided, in time, with very high climatic temperatures recorded in the region. The harvest effect was also observed during the first week of the study year. Establishing appropriate risk signal thresholds is a limiting factor of predictive models; it needs to be adjusted based on experience gained during the use of the models. To increase the sensitivity and specificity of the predictions epidemiological interpretation of non-specific risk signals should be complemented by other sources of information. The methodology developed in this study can enhance other existing early detection surveillance systems. Syndromic surveillance based on mortality monitoring can reduce the detection time for certain disease outbreaks associated with mild mortality only detected at regional level. The methodology can be adapted to monitor other parameters routinely collected at farm level which can be influenced by communicable diseases.

Preclinical safety profile of plasma prepared using the INTERCEPT Blood System.

BACKGROUND AND OBJECTIVES: The newly developed INTERCEPT Blood System for plasma uses the addition of a new psoralen, amotosalen HCl (AMOTOSALEN), followed by illumination with ultraviolet A light, to inactivate viruses, bacteria, protozoa and leucocytes that may contaminate fresh-frozen plasma (FFP). Extensive toxicology studies were performed to characterize the safety of the photochemical treatment process for its intended use with plasma. MATERIALS AND METHODS: The studies of general toxicology, safety pharmacology, phototoxicity, reproductive toxicity and venous irritation, summarized in this review, provide a comprehensive toxicology profile for photochemically treated 100% plasma. RESULTS: No specific target organ toxicity (based on clinical or histological pathology), phototoxicity, or reproductive toxicity was observed. CONCLUSIONS: The results of an extensive series of studies have demonstrated no toxicologically relevant effects of photochemically treated 100% plasma prepared using the INTERCEPT Blood System for plasma.

Preclinical safety of a nucleic acid-targeted Helinx compound: a clinical perspective.

Helinx technology (Cerus Corp, Concord, CA) uses amotosalen HCl (S-59) and ultraviolet A (UVA) light in an ex vivo photochemical treatment (PCT) to inactivate viruses, bacteria, and leukocytes in platelet concentrates while preserving therapeutic function. A comprehensive preclinical safety program was conducted, which included carcinogenicity, single-dose and multiple-dose (up to 13 weeks' duration) toxicity, safety pharmacology (central nervous system [CNS], renal, and cardiovascular), reproductive toxicity, genotoxicity, vein irritation, phototoxicity, and toxicokinetic testing. The results of the toxicokinetic analyses indicated that the test articles provided large multiples of the clinical exposure to S-59, whether the comparison was based on dose, maximum plasma concentration, or area under the concentration-time curve. No specific target organ toxicity, reproductive toxicity, or carcinogenicity was observed. S-59 and/or PCT formulations demonstrated CNS toxicity, electrocardiographic (ECG) effects, and phototoxicity at supraclinical doses. On the basis of the extremely large safety margins, the CNS and ECG observations (at >30,000-fold the expected clinical exposure) as well as the results of genotoxicity and phototoxicity studies are not considered to be of toxicological relevance. The results of an extensive series of studies have thus demonstrated no toxicologically relevant effects of platelets treated with Helinx technology.

The genotoxicity profile of atorvastatin, a new drug in the treatment of hypercholesterolemia.

While HMG-CoA reductase inhibitors such as fluvastatin, lovastatin, pravastatin and simvastatin demonstrate lack of in vitro and in vivo mutagenicity and clastogenicity in bacterial and mammalian cells, long term rodent carcinogenicity studies resulted in an increased incidence in neoplasms at high doses. These effects may be attributable to an exaggeration of the desired biochemical effect of the drug and/or a tumor promoting effect. The genotoxicity of atorvastatin, a newly developed HMG-CoA reductase inhibitor, was evaluated in a variety of test systems. In bacterial mutagenicity tests, the E. coli tester strain WP2(uvrA) and S. typhimurium strains TA98, TA100, TA1535, TA1537, and TA1538 were exposed to concentrations of atorvastatin as high as 5000 micrograms/plate both in the absence (S9-) and presence (S9+) of metabolic activation. Atorvastatin was not mutagenic in either E. coli or S. typhimurium. Chinese hamster lung V79 cell cultures were exposed to atorvastatin at concentrations of 50-300 micrograms/ml (S9-) and 100-300 micrograms/ml (S9+) and structural chromosome aberrations were assessed. Mutation at the hgprt locus was assessed at concentrations of 100-300 micrograms/ml (S9-) and 150-275 micrograms/ml (S9+). Atorvastatin was neither mutagenic nor clastogenic in the absence or presence of S9. The lack of in vitro genotoxicity was corroborated in vivo in a mouse micronucleus study in which single oral doses of atorvastatin were administered to male and female CD-1 mice at 1, 2500, or 5000 mg/kg. No biologically significant increases in the frequency of micronucleated polychromatic erythrocytes in bone marrow at 24, 48, or 72 h postdosing were observed. Thus, atorvastatin, as with the other tested HMG-CoA reductase inhibitors, is not genotoxic.

Hyperbaric oxygen as prophylaxis or treatment for radiation myelitis.

This animal study was designed to investigate HBO as a treatment or prophylaxis for radiation myelitis. All animals received identical spinal cord radiation doses of 69 Gy in 10 daily fractions. Group I received no HBO; group II began HBO at the onset of signs of myelitis; group III received HBO with prophylactic intent beginning 6 wk after irradiation; and group IV received both modalities on the same day, but radiation always preceded HBO by at least 4 h. HBO consisted of 90 min oxygen at 2.4 atm abs for 20 daily treatments. Animals were objectively assessed for the loss of certain neurologic reflexes indicative of four levels of myelitis. Although all animals progressed to severe myelitis, group III animals had group-averaged levels of myelitis consistently less than control. The differences were statistically significant for several weeks. Group IV animals progressed to severe myelitis much more rapidly than any other group. Additional study is justified by this trial. Key questions to be answered include the optimal timing of HBO to produce a beneficial rather than detrimental effect.

High capacity in vitro micronucleus assay for assessment of chromosome damage: results with quinolone/naphthyridone antibacterials.

A high capacity in vitro micronucleus assay was developed to evaluate the ability of selected 6-fluorinated quinolone and naphthyridone antibacterial compounds to induce micronuclei (MN) in vitro in V79 Chinese hamster lung cells. Log-phase cells in six-well cluster dishes were exposed for 3 h in the absence of S9 to 34 compounds. After treatment, cells were refed with media containing cytochalasin B, incubated for 16 h, and harvested for cell-cycle kinetics (CCK) and MN analyses. The quinolones tested were grouped according to the substituent at the 8-position. All 4 compounds having a halogen substitution at position 8, five of the six 8-trifluoromethyl quinolones, and all eight 8-methoxy-substituted compounds induced a significant increase in MN. Only 5 of the 10 naphthyridone compounds tested, having a variety of substituents at the 7-position, were inducers of MN and the overall magnitude of the response was less than with the quinolones. The minimum clastogenic concentration for the quinolones ranged from 4 to 400 micrograms/ml and for the naphthyridones this range was from 22.5 to 100 micrograms/ml. In the groups examined, napthyridone compounds were less likely than quinolones to induce in vitro MN, particularly when the substituent at the 7-position in the naphthyridone contains some bulk (methyl groups) around the amine side-chain. Most of the quinolones tested induced MN, irrespective of the substituents at positions 7 or 8.

Genotoxicity assessment of pirmenol, a new antiarrhythmic drug.

The genotoxicity of pirmenol was tested in the E. coli and S. typhimurium mutagenesis assay, an in vitro mammalian cell chromosome-aberration assay and an in vivo mouse micronucleus assay. The E. coli tester strain WP2s was exposed to concentrations of pirmenol as high as 10,000 micrograms/plate both in the absence (S9-) and presence (S9+) of metabolic activation. Five strains of S. typhimurium (TA98, TA100, TA1535, TA1537, TA1538) were exposed to concentrations of pirmenol as high as 5000 micrograms/plate in the absence and presence of S9. Pirmenol was not mutagenic toward either E. coli or S. typhimurium. Chinese hamster lung V79 cell cultures were exposed to pirmenol at concentrations of 500-2500 micrograms/ml (S9-) and 500-3000 micrograms/ml (S9+). Pirmenol increased the frequency of structural chromosome aberrations (SCAs). The minimum clastogenic concentration was 1500 micrograms/ml (both S9- and S9+) with a peak clastogenic response of 6% (S9-) and 34% (S9+) cells with aberrations. Although there were statistically significant results in the S9- experiment, the percent cells with aberration values for treated groups were within the historical control range (0-6%) of this laboratory. The observed effects in both the absence and presence of S9 appear at high concentrations compared to human circulating plasma levels of 1-3 micrograms/ml and the clastogenicity was confined to chromosome gaps and breaks. Consequently, this in vitro effect would not be expected to be reflected by either in vivo clastogenic or carcinogenic activity. This was supported by findings in the mouse micronucleus study of pirmenol in which single oral doses administered to male CD-1 mice at 5, 55, or 115 mg/kg (80% LD50) produced no statistically significant increases in the frequency of micronucleated polychromatic erythrocytes in bone marrow at 24, 48 or 72 h postdosing. Additionally, no evidence of carcinogenicity was seen in a mouse or rat bioassay.

Simultaneous micronucleus and chromosome aberration assessment in the rat.

Using a cellulose column fractionation procedure to eliminate nucleated cells for micronucleus assessment, micronucleus and chromosome aberration endpoints in the same animal were compared in male and female rats following i.p. injection with cyclophosphamide (CP). Groups of 5 Wistar rats per sex were given single doses of CP at 0, 20, or 40 mg/kg. Two hours prior to sacrifice, animals were given colchicine (4 mg/kg) to arrest cells in metaphase. One femur from each animal was used for micronucleus assessment and the other for chromosome aberration assessment. In the micronucleus assessment, 2000 polychromatic erythrocytes (PCEs) per animal and in the chromosome aberration assessment, 50 metaphase cells per animal were scored. This experiment was repeated once. In both experiments, significant increases in micronucleated PCEs and chromosome aberrations were noted at both doses of CP in both sexes. In general, the clastogenic effects of CP were more pronounced in males than females. Both doses of CP caused a decrease in the proportion of PCEs and in mitotic index in both experiments, indicating toxicity of CP to the bone marrow. These results show the usefulness of this rat model for simultaneous evaluation of two cytogenetic endpoints in the same animal and indicate that assessment of MNPCE frequency in the bone marrow of male rats may be an appropriate model for screening test substances for in vivo clastogenic activity in this species.

Absence of a synergistic effect between moderate-power radio-frequency electromagnetic radiation and adriamycin on cell-cycle progression and sister-chromatid exchange.

In our laboratories we are conducting investigations of potential interactions between radio-frequency electromagnetic radiation (RFR) and chemicals that are toxic by different mechanisms to mammalian cells. The RFR is being tested at frequencies in the microwave range and at different power levels. We report here on the 1) ability of simultaneous RFR exposures to alter the distribution of cells in first and second mitoses from that after treatment by adriamycin alone, and 2) on the ability of simultaneous RFR exposure to alter the extent of sister chromatid exchanges (SCEs) induced by adriamycin alone. This chemical was selected because of its reported mechanism of action and because it is of interest in the treatment of cancer. In our studies, Chinese hamster ovary (CHO) cells were exposed for 2 h simultaneously to adriamycin and pulsed RFR at a frequency of 2,450 MHz and a specific absorption rate of 33.8 W/Kg. The maximal temperature (in the tissue-culture medium) was 39.7 +/- 0.2 degrees C. The experiments were controlled for chemical and RFR exposures, as well as for temperature. Verified statistically, the data indicate that the RFR did not affect changes in cell progression caused by adriamycin, and the RFR did not change the number of SCEs that were induced by the adriamycin, which adriamycin is known to affect cells by damaging their membranes and DNA.

Time lapse and microscopic examinations of insonated in vitro cells.

A fibroblast cell line (Balb/c 3T3) was sham/exposed to ultrasound (2.2 MHz resonant frequency, 35.4 W/cm2 SPTP, 3 microseconds pulse duration, 200 Hz prf) for 30 min and subsequently scored for cell motility and alterations in cell morphology. Cell motility was assessed by computerized tracking of cells. Cell morphology ("normal" or "abnormal") was determined blindly by three independent scorers. A positive control (1-4 Gy) yielded statistically significant alterations in cell motility and morphology; no such differences were observed for the ultrasound regimen. The results thus fail to confirm an earlier report by Liebeskind et al. (Brit. J. Cancer 45(Suppl. V):176-186; 1982) of ultrasound-induced alterations in cell motility and morphology.

Effect of fetal exposure to ultrasound on the development of functional, antigen-specific B lymphocytes in fetal and neonatal BALB/c mice.

In this study, the influence of fetal exposure to ultrasound on the development of immunocompetence is addressed. Pregnant BALB/c mice were exposed to various intensities of ultrasound ranging from 0.1-3.0 W/cm2. B cells from 19 day old fetal livers or 5 and 10 day old neonatal spleens were assessed for the following: (a) differentiation into plasma cells after mitogenic stimulation with lipopolysaccharide (LPS); (b) development and frequency of 2,4-dinitrophenyl (DNP)-specific B cells, and (c) the ability to produce antibody in response to DNP. Comparison between ultrasound-exposed and sham-treated groups did not reveal any evident differences in the above tests. The results suggest that ultrasound neither hinders nor augments the development of immunocompetent B cells.

Effect of fetal exposure to ultrasound on B lymphocyte function and antibody class production.

The purpose of this study was to determine if ultrasound radiation would affect the ability of antigen-stimulated B cells and their clonal progeny to undergo the isotype switch and produce diverse antibody classes. Pregnant BALB/c mice were exposed to various intensities of ultrasound (0.1-3.0 W/cm2) at different ages of gestation. Spleens from the resulting offspring were removed five or ten days after birth. The splenocytes were analyzed for the frequency of B cells capable of responding to the hapten 2,4 dinitrophenyl (DNP) using the splenic fragment assay, a B cell cloning assay. The DNP-responsive B cell clones were then classified on the basis of isotype expression. It was found that ultrasonic radiation during gestation did not alter the frequency of B cells responding to DNP. Furthermore, ultrasound had no apparent effect on the B lymphocyte's capacity to switch to different isotypes after antigenic stimulation. Thus, the results indicate that prenatal exposure to ultrasound does not appreciably affect the genetic and cellular processes necessary for the isotype switch and antibody class production.

Effect of fetal exposure to ultrasound on B cell development in BALB/c mice.

Ultrasound is a major tool for clinical diagnosis, especially during prenatal life. Therefore, it is important that the potential bioeffects of ultrasound be determined. In this report, the effects of ultrasound on the development of B lymphocytes is studied. Pregnant BALB/c mice were exposed to intensities of ultrasound ranging from 0.1-3.0 W/cm2, receiving either a single exposure on day 11, or multiple exposures on days 14, 15, and 18 or 19 of gestation. Fetal livers were removed on day 19 of gestation whereas spleen cells were obtained from 5 and 10 day old neonates. These cell populations were analyzed for: (a) the frequency of B220+ B lineage cells; (b) the frequency of immunoglobulin-positive (Ig+) cells, and (c) the ability to proliferate in response to the B cell mitogen lipopolysaccharide (LPS). None of the tests performed revealed any substantial differences between ultrasound-exposed versus sham-treated control animals. In addition, the development of blood cell types other than B lineage cells remained unaffected by exposure to ultrasound. Therefore, exposure to ultrasound at the intensities used does not appear to hinder hemopoiesis or the normal development of B lymphocytes during fetal and early neonatal life.

DSA in acute gastrointestinal hemorrhage: clinical and in vitro studies.

Selective intraarterial digital subtraction angiography (DSA) was used to examine 37 patients with acute gastrointestinal (GI) tract bleeding. Conventional screen-film angiography was used as an adjunct to DSA when a larger field of view was needed (five patients) and when bowel motion prevented the acquisition of adequate image quality with DSA (two patients). Conventional angiography was also performed in all cases in which there were negative DSA examinations. DSA reduced the mean examination time considerably (20% reduction overall), especially for cases involving embolization therapy (35% reduction). DSA was especially valuable in the upper GI tract, where it was used to rapidly locate and/or assist in the embolization of bleeding sites in 19 of 20 patients with positive angiograms. There were 12 true-negative DSA examinations and one false-negative examination due to the limited field of view (9 inches [22.9 cm]). Bowel and respiratory motion were not important problems in the upper GI tract. In the lower GI tract, the usefulness of DSA was severely limited by the small field of view and the misregistration artifact caused by bowel motion. In an in vitro study, DSA and conventional angiography were compared as to their ability to depict several rates of extravasation of contrast material in a model of GI bleeding. DSA tended to be more sensitive for the detection of simulated extravasation (P less than .07).

Weak low frequency electromagnetic fields and chick embryogenesis: failure to reproduce positive findings.

Fertilised chicken eggs were incubated for 48 hours while exposed to pulsed trains of magnetic fields having a duration of 0-5 ms, a rise time of 42 microsecond, and a pulse repetition rate of 100 at a magnetic field flux density of 1 microT. Some eggs were exposed to 1,552 rad X-rays as a positive control. After exposure the embryos were scored blind for a variety of morphological features. X-irradiated eggs displayed highly significant and repeatable anatomical alterations. There were no differences between magnetic field-exposed, sham-exposed and control eggs.

Effects of radiofrequency radiation and simultaneous exposure with mitomycin C on the frequency of sister chromatid exchanges in Chinese hamster ovary cells.

Chinese hamster ovary (CHO) cells were exposed for 2 hr with and without mitomycin C (MMC) (1 X 10(-8)M) to pulsed wave radiofrequency radiation (RFR) at 2450 MHz. The repetition rate of 25,000 pulses per sec (pps), pulse width of 10 microseconds, and exposure geometry used, resulted in a specific absorption rate (SAR) of 33.8 W/kg. The following exposure regimens were used: a 37 degrees C water bath control; a water bath temperature control (TC) in which the continuously monitored medium temperature closely followed the temperature rise in the RFR-exposed flasks; and the RFR-exposed cells in a water bath set at 37 degrees C prior to exposure. RFR exposure resulted in a maximum cell culture medium temperature of 39.2 degrees C. In the absence of MMC, there was no significant increase in sister chromatid exchange (SCE) in the RFR-exposed or TC groups over that of the 37 degrees C control. When a simultaneous treatment of RFR and MMC occurred there was no statistical difference in SCE frequency from that caused by chemical treatment alone.

Sister-chromatid exchanges in human lymphocytes exposed in vitro to therapeutic ultrasound.

Human lymphocytes were exposed in vitro to therapeutic levels of ultrasound (1 W/cm2, CW, 0.87 MHz, durations of 80 and 160 sec). There were no significant differences in sister-chromatid exchange frequencies between controls and ultrasound-exposed cells. Exposure of lymphocytes to the positive control (mitomycin C) resulted in a significant increase in sister-chromatid exchanges. The data do not verify a report by Stella et al. (Mutation Res., 138 (1984) 75-85) that such exposures result in increased frequencies of SCEs.

Factors modifying contrast media induced fibrillation.

Factors effecting contrast media induced ventricular fibrillation were studied in anesthetized dogs using contact time as the measured parameter. Injections of meglumine/sodium diatrizoate (370 mg I/ml) were made into the right coronary artery at 0.4 ml/s until fibrillation occurred. A contrast medium containing calcium chelators was found to produce fibrillation in a significantly shorter contact time than a similar medium without calcium chelators. Pre-treatment by the cardiac glycoside, ouabain, increased the contact time for fibrillation as did the production of a sub-acute infarction in the left coronary artery and a previous fibrillation and resuscitation. The data suggest that calcium binding additives increase the risk of fibrillation while pre-treatment with cardiac glycosides, the presence of stable infarcts in the non-injected areas, or a previous fibrillation and resuscitation do not increase risk.

Lack of effect of high-intensity pulsed ultrasound on sister chromatid exchange and in vitro Chinese hamster ovary cell viability.

The frequency of sister chromatid exchanges (SCEs) in in vitro Chinese hamster ovary cells and their viability were not affected by 3-min exposures to 2-4 MHz, focused, pulsed ultrasound with a pulse repetition rate of 200 Hz, pulse duration of 10 mu sec and intensities (SPTP) of 500 W/cm2 and 2500 W/cm2. The viability results are consistent with those reported elsewhere; the SCE response does not verify a specific previously reported positive response.

Diagnostic ultrasound and sister chromatid exchanges: failure to reproduce positive findings.

Human lymphocytes were exposed in vitro to ultrasound from two clinical devices, one of which was previously reported to have increased the frequency of sister chromatid exchanges. The ultrasonic exposures had no significant effect on the frequency of sister chromatid exchanges from three blood donors. Exposure to ultrasound also had no effect on cell cycle progression. A concomitant positive control (mitomycin C) resulted in a significant increase in sister chromatid exchanges.