Development of MAPT S305 mutation models exhibiting elevated 4R tau expression, resulting in altered neuronal and astrocytic function.

Due to the importance of 4R tau in the pathogenicity of primary tauopathies, it has been challenging to model these diseases in iPSC-derived neurons, which express very low levels of 4R tau. To address this problem we have developed a panel of isogenic iPSC lines carrying the MAPT splice-site mutations S305S, S305I or S305N, derived from four different donors. All three mutations significantly increased the proportion of 4R tau expression in iPSC-neurons and astrocytes, with up to 80% 4R transcripts in S305N neurons from as early as 4 weeks of differentiation. Transcriptomic and functional analyses of S305 mutant neurons revealed shared disruption in glutamate signaling and synaptic maturity, but divergent effects on mitochondrial bioenergetics. In iPSC-astrocytes, S305 mutations induced lysosomal disruption and inflammation and exacerbated internalization of exogenous tau that may be a precursor to the glial pathologies observed in many tauopathies. In conclusion, we present a novel panel of human iPSC lines that express unprecedented levels of 4R tau in neurons and astrocytes. These lines recapitulate previously characterized tauopathy-relevant phenotypes, but also highlight functional differences between the wild type 4R and mutant 4R proteins. We also highlight the functional importance of MAPT expression in astrocytes. These lines will be highly beneficial to tauopathy researchers enabling a more complete understanding of the pathogenic mechanisms underlying 4R tauopathies across different cell types.

Chlamydia gallinacea: a widespread emerging Chlamydia agent with zoonotic potential in backyard poultry.

Chlamydia gallinacea, a new chlamydial agent, has been reported in four European countries as well as Argentina and China. Experimentally infected chickens with C. gallinacea in previous study showed no clinical signs but had significantly reduced gains in body weight (6·5-11·4%). Slaughterhouse workers exposed to infected chickens have developed atypical pneumonia, indicating C. gallinacea is likely a zoonotic agent. In this study, FRET-PCR confirmed that C. gallinacea was present in 12·4% (66/531) of oral-pharyngeal samples from Alabama backyard poultry. Phylogenetic comparisons based on ompA variable domain showed that 16 sequenced samples represented 14 biotypes. We report for the first time the presence of C. gallinacea in North America, and this warrants further research on the organism's pathogenicity, hosts, transmission, and zoonotic potential.

A rare case of primary epithelioid angiosarcoma of the ureter.

Angiosarcoma is an extremely rare vascular malignancy with a 1-year survival rate of 50%, regardless of tumour origin. Distant metastases are common and occur in the lungs, bone, lymph nodes and soft tissues. The majority of patients with angiosarcoma present with localised disease, although 25-45% have distant metastases at presentation. There are few reported cases of angiosarcomas of the bladder and we report the second case in the literature of primary angiosarcoma of the ureter. We suggest that, in atypical manifestations of suspected urothelial malignancy, the multidisciplinary team considers the possibility of rarer, aggressive tumours early. This may influence clinical decision making towards offering radical treatments earlier, before it is too late to do so owing to aggressive oncological behaviour.

The responses of professional groups to the use of Section 136 of the Mental Health Act (1983, as amended by the 2007 Act) in Gloucestershire.

BACKGROUND: Section 136 (S136) of the Mental Health Act (1983, as amended by the 2007 Act) empowers the police to detain those suspected of being mentally disordered in a public place and to convey them to a place of safety (POS) for further assessment. Gloucestershire has not had a specialist facility for S136 detentions and individuals were taken to the police cells or occasionally A&E departments for assessment. AIMS: This paper forms one part of three aspects under investigation. Two companion papers by the authors describe the use of S136 using anonymised audit data and the experiences of detainees. The objectives of this paper have been to assess the responses of the different professional groups involved in the process of S136. METHOD: An anonymous postal questionnaire was distributed to eight groups of professionals who were identified as having the potential to be involved in part of the process of a S136 detention. Results were collated and analysed, and formed the basis for a series of follow-up focus groups within groups to explore themes that warranted further investigation. RESULTS: An overall response rate of 59% was achieved. Seventy-four per cent of participants thought that there should be an alternative POS to the police station. A&E was thought to be an unsuitable alternative POS, with a psychiatric hospital being the first choice for 58%. CONCLUSIONS: There is a gap in the expectations of the different agencies involved in the S136 process, which have the potential to be divisive if interagency pathways and agreements are not in place.

The use of Section 136 (Mental Health Act 1983) in Gloucestershire.

The small number of published studies on the use of Section 136 (S136) of the Mental Health Act 1983 may reflect neither the current situation nor the true national picture as most of the research comes from large urban centres and was published before 1997. We studied the use of S136 between 2002 and 2006 in Gloucestershire, a rural English county with a population of approximately 585,000, by analysing data held by the police and local mental health services. On average 192 subjects (range 176-203) were detained each year under S136, equating to a population rate of 32.8 S136 detentions per 100,000 per year in Gloucestershire. In contrast to other published studies, ethnic minorities, particularly Afro-Caribbean males, were overrepresented only in a minor way. Of those individuals detained, about a third were admitted, a lower rate than in other studies. Acts or threats of self-harm were common (55%), but acts or threats of violence (28%) and evidence of intoxication (16%) were present in a minority, suggesting that detainees are more likely to pose a risk to themselves than others.

A pilot evaluation of saline sonohysterography for postmenopausal bleeding with thickened endometrium.

The aim of this study was to assess the clinical and cost implications of pilot introduction of saline sonohysterography for postmenopausal bleeding with thickened endometrium (>or= 5 mm), at the Royal Glamorgan Hospital. The investigation was attempted in 48 patients who were found to have a thickened endometrium (>or= 5 mm) on transvaginal ultrasound scanning. The attempt was successful in 45 patients (93.8%). Failures were due to (1) cervical stenosis and (2) leakage of saline from the cervix. Difficult catheterisation of the cervix causing pain was encountered in a minority of cases. Two-thirds of patients were found have focal lesions and were booked for hysteroscopic resection. These patients included 30 with polyps, which were precisely described in all of them. In view of this low complication rate and high accuracy, the technique was integrated as a standard method of investigation for postmenopausal bleeding at the hospital.

Hyper-Rayleigh scattering as a means of monitoring crystal nucleation in solution.

Hyper-Rayleigh scattering is revealed as a very sensitive monitor of cluster formation in solution, and as a means of studying the mechanism of crystal nucleation in molecular species. Two compounds are selected with particularly high second harmonic generation (SHG) powers in the crystalline state and experimental conditions are defined allowing the measurement of the beta value for one of these as 18+/-1x10(-30) esu. It is found to agree with current theoretical prediction of 20x10(-30) esu. In the more powerful of these, two photon induced fluorescence is found to be partly responsible for the SHG. The solubilities of both compounds in methanol are measured and it is observed that these differ by a factor of ten. When the solution concentration is increased beyond 45% of the saturation value, the quadratic coefficient exhibits non-linear behaviour with respect to concentration. Additionally, the widths of the distributions of the HRS signals increase initially with concentration as expected, but, beyond 45% saturation concentrations, these narrow again. These phenomena are interpreted as indicators of cluster formation in these solutions well below saturation concentrations. A future experimental design is proposed in which the coherent component will yield information on the organisation of the molecules in such clusters.

Pharmacokinetics of ceftiofur in llamas and alpacas.

Pharmacokinetic studies of antibiotics in South American camelids are uncommon, therefore drugs are often administered to llamas and alpacas based on dosages established in other domestic species. The disposition of ceftiofur sodium was studied in llamas following intramuscular administration and in alpacas following intravenous and intramuscular administration. Eleven adult llamas were given ceftiofur sodium by intramuscular injection. Each animal received either a standard dose of 2.2 mg/kg or an allometrically scaled dose ranging from 2.62 to 2.99 mg/kg in a crossover design. Ten adult alpacas were given ceftiofur sodium by intravenous and intramuscular injections. Each animal received a standard dosage of 1 mg/kg or an allometrically scaled dose ranging from 1.27 to 1.44 mg/kg i.v., and 1.31-1.51 mg/kg i.m. Blood samples were collected at 0, 0.25, 0.5, 1, 2, 4, 8, 12, 24, 36, 48, and 72 h after administration of the ceftiofur. Pharmacokinetic parameters of ceftiofur in llamas and alpacas were similar following i.m. dosing at both dose levels. The only differences noted were in the total AUC between dose levels, but the AUC/dose values were not different. A sequence effect was noted in the alpaca data, which resulted in lower AUCs for the second dose when the i.v. dose was given first, and with higher AUCs for the second dose when the i.m. dose was given first. Overall, ceftiofur pharmacokinetics in llamas and alpacas are similar, and also very similar to reported parameters for sheep and goats.

Leptin, tumor necrosis factor-alpha (TNF), and CD14 in ovine adipose tissue and changes in circulating TNF in lean and fat sheep.

Four studies were designed to determine whether 1) tumor necrosis factor-alpha (TNF) and the Lipopolysaccharide (LPS) binding ligand, CD14, are produced by sheep adipose tissue; 2) nutritional reserves and/or short-term fasting affect circulating concentrations of TNF; 3) there is a relationship between TNF and metabolic factors in sheep; and 4) inflammation alters circulating concentrations of leptin. In Exp. 1 and 2, ewes were assigned, based on ultrasonic assessments of last-rib subcutaneous fat measurements to fat (fat thickness > 1 cm; mean = 1.52 +/- 0.03 cm) or thin (fat thickness < 1 cm; mean = 0.25 +/- 0.03 cm) groups. Fat and thin ewes were assigned to fed or fasted groups for a total of four groups (fed-fat; fasted-fat; fed-thin; fasted-thin). Fed-ewes had ad libitum access to feed, and fasted-ewes were prohibited feed 48 h before initiation of sample collection. In Exp. 1, subcutaneous fat samples were collected from just above the last rib for detection of TNF and CD14 mRNA, and immunoreactivity. Tumor necrosis factor-alpha-like immunoreactivity in adipocytes was sparse, more pronounced in cells in fed-ewes than fasted-ewes, and localized to membranes between adjacent cells in nucleated regions. Immunoreactivity for CD14 was minimally observed but present in adipocytes and widely expressed in infiltrating monocytes and epithelial vascular cells. Leptin was detected in adipocytes. In Exp. 2, plasma samples collected every 6 h for 24 h were analyzed for plasma concentrations of TNF. Fat ewes had greater plasma concentrations of TNF than thin ewes (P = 0.039). In Exp. 3, wethers were injected i.v. with interleukin-1beta or TNF. Blood samples were collected every 15 min for 8 h following injection. Plasma concentration of leptin was not affected by treatment (P > 0.39). In Exp. 4, wethers were injected with LPS. Blood samples were collected every 15 min for 8 h following injection. Plasma concentration of leptin was not altered by LPS (P > 0.20). These results provide evidence: 1) of TNF-like immunoreactivity within fat tissue; 2) that elements within fatty tissues have CD14 that may allow adipocyte function to be directly affected by LPS; 3) that plasma concentrations of leptin are not altered by LPS treatment; and 4) that circulating concentrations of TNF are elevated with obesity in sheep.

Evaluation of the safety and efficacy of testicular biopsies in llamas.

Evaluation of the reproductive function of Lama glama is generally considered to be a challenging task due to the difficulty of obtaining representative semen samples. One method that has been proposed for evaluation of testicular function in these animals is histologic examination of testicular needle biopsies. This study was undertaken to examine the safety and efficacy of using needle biopsies to assess testicular function in this species. One randomly selected testicle from each of 16 sexually mature llamas was biopsied with a 14-gauge self-firing biopsy instrument. The llamas were evaluated over a 6-week period with thermography for temperature changes of the scrotum. At the end of the 6-week trial, the llamas were castrated and sections of each testis were fixed in Bouin's solution for histologic examination. Immediately prior to castration, an additional biopsy was taken from each testis to compare the tissue obtained via biopsy with sections from the corresponding testis obtained after castration. A qualitative grading scale was used to compare the seminiferous tubules from each testis. No difference was found between the biopsied and the nonbiopsied testes (P = 0.69). The percentage of normal tubules between the biopsied and the nonbiopsied sides also did not differ (P = 0.70). Furthermore, the percentage of normal seminiferous tubules did not differ between the needle biopsy samples and the corresponding tissue samples obtained at castration (P = 0.48). The number of round seminiferous tubules counted in each biopsy section ranged from 3 to 67. There was no significant difference in the thermographic images of the scrotum between the biopsied and the nonbiopsied testes. This study supports testicular biopsies as a safe and useful procedure in the evaluation of testicular function.

The EU precautionary bans of animal feed additive antibiotics.

Toxicologists, with good reason, will feel that the biological safety of chemical products across the market sectors rests largely on their efforts. However, one sector has received much adverse attention from the media, consumers, politicians, legislators and advisory groups in recent years. It is food animal production in intensive systems and, within those, various types of chemical additives included in the compound diets fed. No additive class received more adverse comment than those antibiotics used for the purpose of enhancing the efficiency of animal production. This paper considers the safety of the antibiotic feed additives (AFAs) against the background of the regulatory measures in place, defines their role and describes the relevant concerns. It closes with comment on the microbiologically-based health risk which underpinned the AFA bans and sounds a warning over the precedent created by the use of the precautionary principle in the recent banning of six of their number.

Internal parasite control strategies.

To maintain the general health of small ruminants, owners, managers, and clinicians should be familiar with the body condition-scoring [table: see text] methods, general health, and overall appearance of the animals in their care. Any changes in the overall health of individual animals or the flock should be noted immediately, the cause determined, and immediate measures should be taken to correct any problems. Parasite management in small ruminants is a difficult and arduous task. When a program that reduces parasite infestation is discovered, it should be carefully implemented, maintained with vigilance, and constantly "tweaked" to maximize its effectiveness.

A comparison of physical examination and clinicopathologic parameters between sheared and nonsheared alpacas (Lama pacos).

The purpose of this study was to determine the physiological changes associated with chronic heat stress in sheared versus nonsheared alpacas. Fourteen intact male adult alpacas were randomly assigned to one of the two groups: Group S alpacas were sheared to within 2cm of their skin; Group NS alpacas were not sheared. These animals were maintained from June through August in east central Alabama. Data collected in the morning, every two weeks, included vital signs, body weight, body condition score, complete blood counts, serum chemistries and electrolytes, whole blood selenium, and plasma cortisol. S and NS groups were contrasted using the repeated measures analysis of variance, and pertinent correlations with weather parameters were calculated. Clinical heat stress was not evident in any animals during the study. Significant differences between treatment groups were seen in rectal temperature (P=0.0095), sodium concentration (P=0.0219), and blood urea nitrogen (BUN) (P=0.0189). The mean rectal temperature of the NS group was above the normal range on five sampling times compared to only once for the S group. However, mean sodium and serum urea nitrogen levels were within normal limits in both groups at all sampling times. Rectal temperature of only the S group was positively correlated to weather parameters. Sodium of both S and NS groups and BUN of the NS group were negatively correlated with weather parameters. This study indicates that there are differences between sheared and nonsheared alpacas in physical examination and clinicopathologic parameters that can be correlated with changes in ambient conditions. These differences suggest that nonsheared alpacas are less heat tolerant than sheared alpacas. Therefore, shearing is recommended for animals exposed to similar conditions.

A comparison of surface and rectal temperatures between sheared and non-sheared alpacas (Lama pacos).

The objective of this research was to determine if whole-body shearing would effect gross thermoregulation in alpacas. Eight mature, intact male alpacas were randomly assigned to one of two groups and maintained in outdoor pastures with adequate artificial shade from June through August (summer climate) in east central Alabama, USA. Group one animals (N=4) were sheared to remove all fiber to within 2cm of their skin. Group 2 animals (N=4) were left non-sheared. Sheared alpacas tended to have lower rectal temperatures during high ambient temperatures than did non-sheared alpacas (P=0.06). Thermographic studies of the scrotum revealed cooler surface temperatures in sheared versus non-sheared alpacas (P=0.05). Temperatures in the right medial thigh of sheared animals were 0.9 degrees C cooler than the thigh region of non-sheared animals in the morning (P<0.03). Right medial thigh temperatures were 1.6 degrees C cooler in sheared alpacas in the afternoon (P<0.01). Significant positive correlations were found in non-sheared animals between ambient temperature and rectal temperature in the morning (r=0.612, P=0.014). In sheared animals during the morning significant positive correlations were established between the Heat Stress Index (HSI) and the right medial thigh surface temperatures (r=0.648, P=0.003), the HSI and rectal temperature (r=0.729, P=0.0003), the ambient temperature and right medial thigh surface temperature (r=0.485, P=0.04), and the ambient temperature and the rectal temperature (r=0.823, P<0.0001). In the afternoon a significant positive correlation was found in the sheared alpacas between the HSI and the right medial thigh surface temperature, rectal temperature and surface scrotal temperature (r=0.538, P=0.02, r=0.543, P=0.019 and r=0.522, P=0.045), respectively. These data indicate that whole-body shearing of alpacas could have a beneficial effect on thermoregulation when used as a preventative measure against heat stress. Shearing may assist heat dissipation resulting in a cooler surface body temperature and rectal temperature in alpacas when challenged by the heat and humidity experienced in the summer months in the southeastern United States.

Oral treatment with a nutraceutical (Cosequin) for ameliorating signs of navicular syndrome in horses.

Fourteen horses with a progressive forelimb lameness of 3 to 12 months' duration, diagnosed as navicular syndrome, were selected from clinical cases admitted to Auburn University Equine Hospital for evaluation of the efficacy of an orally administered nutraceutical (Cosequin, Nutramax Laboratories, Inc., Edgewood, MD) for ameliorating clinical signs associated with naturally occurring navicular syndrome. Horses were randomly allocated to treatment with the nutraceutical or a placebo. Treatment was five scoops (16.5 g) of powder twice daily in the feed. The test group (n = 8) received a patented nutraceutical consisting of 9 g of FCHG49 (a highly purified glucosamine HCl), 3 g of TRH122 (a specific purified low-molecular-weight sodium chondroitin sulfate), and 600 mg of manganese ascorbate. The placebo group (n = 6) received an indistinguishable oral powder containing only excipients. Owners and the investigator were unaware of group assignments. The same investigator assessed lameness and overall clinical condition at enrollment and after 4 and 8 weeks of treatment. Lameness was assessed by an algofunctional lameness index, comprising a combined sum score of standing posture, hoof tester examination, and lameness scores at various levels of work. Overall clinical efficacy was rated on a visual analogue scale. Owners assessed lameness via a preassigned questionnaire, incorporating an algofunctional lameness index and overall clinical condition at weekly intervals. Radiographic examinations of the navicular bones were performed at enrollment and after 8 weeks of treatment. The median algofunctional lameness index and overall clinical condition scores assigned the investigator were significantly improved (P = .05) for horses treated with the nutraceutical compared with placebo-treated horses. The degree of improvement in algofunctional lameness index assigned by owners after 8 weeks was also significant (P = .045) between the treatment groups. Radiographic scores after treatment were not significantly different between the groups (P > .05).