Veterinary Forensic Pathology of Animal Sexual Abuse.

Animal sexual abuse (ASA) involves harm inflicted on animals for the purposes of human sexual gratification and includes such terms as bestiality, zoophilia, zoosadism, animal sexual assault, and others. The prevalence of ASA is not known, although it may be more common than is currently perceived. Veterinarians have the skills required to identify and document cases of ASA. This article reviews the terminology, legal definitions and forms of ASA, and its social and psychological context. An investigative approach is outlined, including an alternate light source examination; collection of swabs for DNA analysis; sampling vaginal washes, rectal washes, and toenails for trace evidence and biologic analyses; radiographic studies; and a complete forensic necropsy, including histopathology. Gross lesions identified in ASA victims include injuries to the anus, rectum, penis, scrotum, nipples, and vagina; the presence of foreign bodies; and abrasions, bruising, and other evidence of nonaccidental injury. Specialized procedures, including examination using alternate light sources and screening tests to identify human seminal fluid within samples from ASA victims, are of potential value but have not been validated for use in animals.

Viral vector-based improvement of optic nerve regeneration: characterization of individual axons' growth patterns and synaptogenesis in a visual target.

Lack of axon growth ability in the central nervous system poses a major barrier to achieving functional connectivity after injury. Thus, a non-transgenic regenerative approach to reinnervating targets has important implications in clinical and research settings. Previous studies using knockout (KO) mice have demonstrated long-distance axon regeneration. Using an optic nerve injury model, here we evaluate the efficacy of viral, RNA interference (RNAi) and pharmacological approaches that target the phosphatase and tensin homolog (PTEN) and signal transducer and activator of transcription-3 pathways to improve long-distance axon regeneration in wild-type mice. Our data show that adeno-associated virus (AAV) expressing short hairpin RNA (shRNA) against PTEN (shPTEN) enhances retinal ganglion cell axon regeneration after crush injury. However, compared with the previous data in PTEN KO mice, AAV-shRNA results in a lesser degree of regeneration, likely due to incomplete gene silencing inherent to RNAi. In comparison, an extensive enhancement in regeneration is seen when AAV-shPTEN is coupled to AAV encoding ciliary neurotrophic factor (CNTF) and to a cyclic adenosine monophosphate (cAMP) analog, allowing axons to travel long distances and reach their target. We apply whole-tissue imaging that facilitates three-dimensional visualization of single regenerating axons and document heterogeneous terminal patterns in the targets. This shows that some axonal populations generate extensive arbors and make synapses with the target neurons. Collectively, we show a combinatorial viral RNAi and pharmacological strategy that improves long-distance regeneration in wild-type animals and provide single fiber projection data that indicates a degree of preservation of target recognition.

Risks posed by climate change to the delivery of Water Framework Directive objectives in the UK.

The EU Water Framework Directive (WFD) is novel because it integrates water quality, water resources, physical habitat and, to some extent, flooding for all surface and groundwaters and takes forward river basin management. However, the WFD does not explicitly mention risks posed by climate change to the achievement of its environmental objectives. This is despite the fact that the time scale for the implementation process and achieving particular objectives extends into the 2020s, when climate models project changes in average temperature and precipitation. This paper begins by reviewing the latest UK climate change scenarios and the wider policy and science context of the WFD. We then examine the potential risks of climate change to key phases of the River Basin Management Process that underpin the WFD (such as characterisation of river basins and their water bodies, risk assessments to identify pressures and impacts, programmes of measures (POMs) options appraisal, monitoring and modelling, policy and management activities). Despite these risks the WFD could link new policy and participative mechanisms (being established for the River Basin Management Plans) to the emerging framework of national and regional climate change adaptation policy. The risks are identified with a view to informing policy opportunities, objective setting, adaptation strategies and the research agenda. Key knowledge gaps have already been identified during the implementation of the WFD, such as the links between hydromorphology and ecosystem status, but the overarching importance of linking climate change to these considerations needs to be highlighted. The next generation of (probabilistic) climate change scenarios will present new opportunities and challenges for risk analysis and policy-making.

Physiotherapy following cardiac surgery: is it necessary during the intubation period?

This randomised controlled clinical trial investigated whether physiotherapy during the period of mechanical ventilation following cardiac surgery influenced subject outcomes. Two hundred and thirty-six subjects admitted to the intensive care unit (ICU) following elective or semi-urgent cardiac surgery were randomised to either a treatment group, which received physiotherapy during the intubated phase, or a control group where physiotherapy was commenced only once the subject was extubated. No significant differences between the two groups were detected for length of intubation period, length of ICU stay, length of hospital stay, maximal daily incentive spirometry values or the incidence of post-operative pulmonary complications. For individuals following routine uncomplicated cardiac surgery, the provision of physiotherapy interventions during the post-operative intubation period does not improve outcomes.

The relationship between size and fecundity in Aedes albopictus.

Recent use of correlates of the intrinsic rate of increase and models to study population dynamics in mosquitoes has emphasized the need for species specific data on the relationship of morphological factors such as body size to life history parameters such as fecundity. Aedes albopictus is of concern because of its continued spread and population growth in the U.S. and the potential for virus transmission. We examined the relationships between pupal wet weight and adult female wing length and between female wing length and fecundity in the first gonotrophic cycle in U.S. strains of Ae. albopictus. There was a significant positive relationship between pupal wet weight and adult wing length. The relationship between wing length and fecundity was also significant and positive, but with lower correlation coefficients and higher variance indicating that other factors also affect fecundity. The regression coefficients for the fecundity-wing length equation did not differ between two strains of Ae. albopictus.

Role of Anopheles quadrimaculatus and Coquillettidia perturbans (Diptera: Culicidae) in the transmission cycle of Cache Valley virus (Bunyaviridae: Bunyavirus) in the midwest, USA.

Midwestern populations of Coquillettidia perturbans (Walker) and Anopheles quadrimaculatus (Say) were tested for their ability to transmit Cache Valley virus (CV), a recognized human and animal pathogen. Field-collected mosquitoes were fed artificial blood meals containing 5.2-6.2 log10 pfu/ml of CV. After 9-23 d at 28 degrees C, 75-93% of blood-fed Cq. perturbans had disseminated infections and 6-62% transmitted the virus to suckling mice. However, when infected with a lower virus titer (3.3 log10 pfu/ml), only 10-36% of the mosquitoes had disseminated infections and 0-10% transmitted the virus to suckling mice. A similar infection rate (21%) was observed in Cq. perturbans fed on viremic (3.2 log10 pfu/ml) hamsters. An. quadrimaculatus were infected (81-100%) by both doses used, with transmission rates ranging from 13-67% after 16-23 d of incubation. Transmission rates for the laboratory strain An. quadrimaculatus SAVANNAH ranged from 20 to 33% after 7-14 d of incubation. Our data show that although An. quadrimaculatus is more susceptible to CV infections than Cq. perturbans, both mosquito species could be involved in the midwestern transmission cycle of the virus.

Rat renal cortical slices: Maintenance of viability and use in in vitro nephrotoxicity testing.

The majority of in vitro toxicology studies employing precision-cut rat renal slices are carried out using simple incubation systems over relatively short periods of time (usually up to 8 hr). We aimed to develop a system for longer-term (up to 24 hr) culture of rat renal slices. The viability of precision-cut rat renal cortical slices was therefore investigated under a variety of incubation conditions. Slices cultured using a dynamic organ culture system were found to be more viable than those cultured in shaken multiwell dishes. This improved viability was evident after only 2 hr of culture (as assessed by the measurement of LDH leakage). An oxygen enriched gaseous environment was found to be important for the maintenance of slice viability using the dynamic organ culture system and conditions were established which permitted the culture of viable slices for periods up to 24 hr. Using these optimized incubation conditions, less than 8% LDH leakage was observed at 24 hr and slices retained 84% of fresh ATP values. Histologically, few signs of slice degeneration were noted until 20 hr of culture. The renal slice system was shown to discriminate between the most toxic and least toxic of a series of cephalosporin antibiotics, based on measurements of slice ATP content.

The mosquitoes of Yellowstone National Park (Diptera:Culicidae).

Twenty-nine species belonging to 4 genera of mosquitoes are reported for Yellowstone National Park. W.Y. (USA). Phenology, local distribution, and ecology of each species are addressed along with taxonomic notes on certain species. Merinithid (nematode) parasitism of some of the park's Aedes species is also discussed.

(1)H NMR spectroscopic studies on the characterization of renal cell lines and identification of novel potential markers of in vitro nephrotoxicity.

Abstract Cell cultures are increasingly used in the evaluation of chemically-induced nephrotoxicity. The utili of renal cell culture systems in toxicology would be improved, however, if better characterized and more specific markers of toxicity were available. High resolution proton nuclear magnetic resonance ((1)H NMR) spectroscopy is well suited to the study of toxicological events and has identified many novel markers of nephrotoxicity in vivo. In this study, (1)H NMR spectroscopy has been used to characterize the biochemical composition of two renal cell lines of different nephronal origin, LLC-PK1 (pig proximal tubule) and Madin-Darby canine kidney (MDCK, distal tubule). The early biochemical responses of these cell lines to the model proximal tubular toxin S-(1,2dichlorovinyl)i-L-cysteine (DCVC) and the renal medullary toxin 2-chloroethanamine (CEA) have also been investigated. For each line, 500 MHz (1)H NMR spectra of protein-free acetone extracts of cells and culture medium gave characteristic and reproducible profiles of low MW constituents, including amino and organic acids, glucose and soluble membrane precursors, such as choline and myo-inositol. Treatment-related changes in several low MW compounds not routinely measured in toxicological studies were revealed by NMR specboscopy before marked cytotoxicity was observed by phase contrast microscopy. For example, LLC-PK1 cells treated with 60 µM DCVC showed a marked decrease in intracellular choline levels within 3 h which suggests an effect on the balance of choline synthesis and utilization. Wrthin 9 h of treatment with DCVC there were decreases in intracellular acetate and alanine concentrations which may be indicative of a decrease in fatty acid oxidation and biglyceride metabolism accompanied by an increase in gluconeogenesis. In MDCK cells, 1 h post treatment with 5 mM CEA, intracellular glycine was decreased. This study indicates the potential power and applicability of (1)H NMR spectroscopy for evaluating the biochemical and metabolic effects of toxins in cell culture systems and provides a novel approach to identifying new markers of tissue damage.

1H NMR spectroscopic studies on the reactions of haloalkylamines with bicarbonate ions: formation of N-carbamates and 2-oxazolidones in cell culture media and blood plasma.

1H NMR spectroscopic methods have been applied to compare the in vitro reactivity of the renal papillary nephrotoxin 2-bromoethanamine (BEA) with those of selected halide-substituted nephrotoxic analogues, 2-chloroethanamine (CEA), 2-fluoroethanamine (FEA), and 1-phenyl-2-iodoethanamine (PIEA). The primary 1H NMR-detectable transformation during a 24 h incubation of confluent Madin Darby canine kidney (MDCK) cells with BEA, CEA, and FEA (at concentrations up to the IC50 determined by neutral red uptake) was the appearance in cell culture media of 2-oxazolidone (OX). Additional novel signals assigned as FEA carbamate (N-carbamoyl-2-fluoroethanamine) were observed in media collected following incubation of cells with FEA. We propose that N-carbamate intermediates are formed from the spontaneous reaction of these haloalkylamines with HCO(3-)-buffered growth media and that OX is formed from the carbamate via elimination of the hydrogen halide. Further 1H NMR experiments, conducted for up to 8 h at 25 degrees C on 5 mM solutions of BEA, CEA, and FEA in 2H2O containing a 20-fold excess of HCO3- at pH 7.6, demonstrated a time-dependent decrease in the concentration of the free haloalkylamines accompanied by the production of N-carbamate intermediates and OX. Under these pseudo-first-order reaction conditions, the formation of OX from BEA was complete within approximately 6 h. In similar reaction conditions OX formation from CEA (24 h after initiation) had reached 54% of its final equilibrium concentration. Equivalent experiments demonstrated that PIEA was almost completely converted to 4-phenyl-2-oxazolidinone (PHOX) within 2 h. These observations reveal the strong disposition of this series of haloalkylamines toward reaction with HCO3- and indicate that the compounds in this family may exist only transiently as free amines in vivo, where there will virtually always be excess HCO3-. The physiological relevance of the in vitro findings is further indicated by the NMR-detectable conversion of BEA to OX and also an alkylating aziridine (AZ) moiety in rat plasma containing BEA. The ability to form carbamoylated species and OX (or PHOX) may mediate the toxicity of this series of haloalkylamines and hence is potentially of considerable significance.

Parasitism of Aedes aegypti and Ae. albopictus (Diptera: Culicidae) by Ascogregarina spp. (Apicomplexa: Lecudinidae) in Florida.

Aedes aegypti (L.) and Ae. albopictus (Skuse) from 40 sites in 17 counties of Florida were surveyed for gregarine parasites during the spring and autumn of 1993 and in July 1994. Larvae collected from containers (mainly tires) were dissected to determine the number of gregarine trophozoites present. Ascogregarina spp. were found at 70% of the sites and occurred as frequently in Ae. aegypti populations as in Ae. albopictus. Within sites, parasite distributions were highly variable and aggregated in host populations. Parasite loads ranged from 1 to 486 trophozoites per host. Mean parasite load was significantly higher in Ae. aegypti larvae (52.5 per host) than in Ae. albopictus (33.5 per host). Parasite prevalence was significantly higher in Ae. aegypti populations that were not sympatric with Ae. albopictus compared with allopatric Ae. albopictus or sympatric populations of either species. In sympatric populations, Ae. aegypti tended to have equal or higher parasite prevalence than the cohabiting Ae. albopictus. Ascogregarina taiwanensis (Lien & Levine) prevalence in Ae. albopictus was significantly higher in areas where these hosts have been present for at least 3 yr. These data contribute to the hypothesis that parasite-mediated competition may be a factor in the apparent displacement of Ae. aegypti by Ae. albopictus in Florida.

Measuring health status in psoriatic arthritis: the Health Assessment Questionnaire and its modification.

OBJECTIVE: The Health Assessment Questionnaire (HAQ) has proven to be a reliable and valid measure of outcome for a variety of arthritides. A recent modification of HAQ for spondyloarthropathy (HAQ-S) has also been reported. Our purpose was to evaluate the HAQ and HAQ-S as outcome measures in the assessment of patients with psoriatic arthritis (PsA). METHODS: The HAQ, including HAQ-S was administered to all patients attending our Psoriatic Arthritis Clinic between June and December, 1993. Clinical and radiological assessments were performed according to a standard protocol that measures disease activity, fibrositic tender points (TP), disease severity and damage. Analysis was performed using SAS for the PC. RESULTS: The patient population included 114 patients, 70 men and 44 women with a mean age of 49.3 years and a mean arthritis duration of 15.1 years. The mean HAQ score was 0.50, while the mean HAQ-S score was 0.53 (scores range 0 to 3 for this instrument). The overall HAQ and HAQ-S disability scores were highly correlated with several clinical measures of function, including grip strength (r = -0.63 and -0.59, respectively). American College of Rheumatology functional class (r = 0.59 and 0.60, respectively), as well as the number of fibrositic TP (r = 0.54 and 0.57, respectively). These disability scores also correlated highly with the overall number of actively inflamed joints (r = 0.49 and 0.50, respectively); however, they correlated only moderately or poorly with other measures of disease activity such as morning stiffness, total number of joint effusions, erythrocyte sedimentation rate (ESR) and the PASI score for psoriasis and with all measures of disease severity. A similar pattern of correlations was found between the individual subscales of the HAQ and HAQ-S and the clinical measures of function, activity, and severity, as well as between the pain scale and the various clinical measures. However, the correlations are generally lower. CONCLUSION: Our data suggest that HAQ and HAQ-S capture clinical measures of function and pain in PsA but do not correlate with disease severity. The HAQ and its modification for spondyloarthropathy may reflect fibromyaglia as a measure of pain and tenderness in these patients. Thus, the clinical assessment of disease activity and both clinical and radiological assessments of joint damage remain important outcome measures in PsA.

Autocrine growth stimulation of human renal Wilms' tumour G401 cells by a gastrin-like peptide.

The role of gastrin in the control of growth of renal G401 cells isolated from a human nephroblastoma (Wilms' tumour) was investigated. G401 cell growth was enhanced in the presence of exogenous gastrin. Addition of anti-gastrin antibodies to serum-free medium significantly inhibited the growth of G401 cells. G401 cells contained the equivalent of 4.3 pg/10(6) cells of gastrin, and serum-free medium collected over 48 hr from G401 cells contained the equivalent of 38 ng/10(6) cells of gastrin, as determined by radioimmunoassay. Growth of G401 cells was inhibited in a concentration-related way by a variety of gastrin/CCK receptor antagonists. Devazepide and proglumide were, respectively, the most and the least potent inhibitors of G401 cell growth (potency order devazepide > L-365,260 = lorglumide > loxiglumide > benzotript > proglumide). These gastrin/CCK receptor antagonists had similar growth-inhibitory activities in human colonic adenocarcinoma HCT-116 cells. Growth of HCT-116 cells was stimulated to a lesser extent, as compared with G401 cells, by exogenous gastrin, and endogenous gastrin was not detectable in HCT-116 cells. The results are consistent with a role for a gastrin-like peptide in the control of growth of a renal cell line. The data suggest that gastrin/CCK receptor antagonists warrant further investigation as therapeutic agents for the control of gastrin-responsive tumours derived from outside, as well as inside, the gastrointestinal tract, including tumours derived from the kidney.

Use of scanning electron microscopy in the study of nephrotoxin-induced renal proximal tubular damage in vitro.

Scanning electron microscopy was used to study cephaloridine and gentamicin-induced renal cell injury in vitro. Exposure of renal proximal tubular cells to these toxicants produced changes in cell surface morphology that occurred at concentrations similar to (or below) those affecting neutral red uptake or monolayer permeability.

In vitro toxicity of cephalosporin and aminoglycoside antibiotics to LLC-PK(1) cells assessed by changes in glucose uptake.

Nephrotoxin-induced damage to cultured kidney cells (LLC-PK(1)) was investigated using assays of general cytotoxicity (neutral red uptake) and of functional impairment (glucose accumulation). The latter correctly ranked both the cephalosporin and aminoglycoside antibiotics (acute and chronic proximal tubule toxicants respectively) in terms of their relative in vivo toxicities. Functional impairment after low-dose long-term exposure to the aminoglycosides occurred at sublethal drug concentrations. Assessment of general cytotoxicity correctly ranked only the aminoglycoside antibiotics.

Records of mosquito-parasitic mermithid nematodes in the northcentral United States.

Mermithid parasites (Nematoda: Mermithidae) were observed in 11 mosquito species in 22 counties of Indiana, Michigan, Minnesota, Ohio and Wisconsin. Natural hosts included adult Aedes vexans, Ae. canadensis, Ae. cinereus, Ae. diantaeus, Ae. punctor, Anopheles punctipennis and Coquillettidia perturbans and larvae of Ae. abserratus, Ae. canadensis, Ae. cinereus, Ae. communis, Ae. diantaeus, Ae. provocans, Ae. punctor, Ae. stimulans and Cq. perturbans. These are the first records of such parasites in Indiana, Minnesota, Ohio and Wisconsin.