How does visible light flicker impact laying hen pullet behavior, fear, and stress levels?

Many characteristics of artificial light have been evaluated; however, light-flicker frequency (F) has not been assessed extensively in poultry. Pullets (1,344 per strain [S]; Lohmann Brown-Lite [LB] and LSL-Lite [LW]) were placed into 8 light-tight rooms, containing 6 floor pens (15 pen replicates per F × S for 30 and 250 Hz; 18 pen replicates per F x S for 90 Hz), and assigned 1 of 3 F treatments (30, 90, 250 Hz). The experiment took place over 2 trials (blocks). To evaluate long-term effects of F during rearing, birds were followed through the hen phase. Data were analyzed using Proc Mixed (SAS 9.4). Differences were considered significant when P ≤ 0.05, and behaviors are expressed as percentage of time. Pullets reared under 30 Hz spent more time performing nutritive behaviors (P < 0.01) and as "unidentified" (P = 0.02) than other treatments. Active behavior demonstrated an age x F interaction, with pullets being more active at 16 wk, regardless of F (P < 0.01). Comfort behaviors were higher at 16 wk compared to other ages, regardless of F (P < 0.01). Exploratory behaviors were lowest at 4 wk in pullets under 30 Hz (P < 0.01). Aggressive behaviors (12 wk) were higher in pullets reared under 250 Hz than those under 90 Hz (P < 0.01). Comb score was unaffected by F (P = 0.79) and all birds scored had a full plumage. Heterophil-to-lymphocyte ratio was unaffected by F at 7 or 15 wk (P = 0.85 and P = 0.54, respectively). In trial 1, pullets reared under 90 Hz had higher corticosterone concentrations than those reared under 250 Hz (P = 0.02) and trial 2 there were no effects of F (P = 0.97). For novel object test, LW pullets reared under 90 Hz had a higher latency to peck than LW pullets under 30 Hz or 250 Hz (P = 0.03). Hen behavior (wk 39) and fear tests (36 wk; novel object test (P = 0.86) and tonic immobility (P = 0.37)) were unaffected by F. Overall, minimal effects of F were seen on pullet and hen behavior and stress.

Do flickering LED lights reduce productivity of layer pullets and hens?

Most characteristics of artificial light sources are well studied, however light-flicker frequency (F) has been overlooked. The purpose of this study was to determine the effect of F on performance of Lohmann LSL-Lite (LW) pullets and Lohmann Brown-Lite (LB) pullets. In addition, pullets were followed through to the laying phase to evaluate long-term effects of F during rearing on productivity. Two trials were conducted with 3 F (30, 90, or 250 Hz) treatments. LW and LB pullets (n = 2,688 per strain [S]) were randomly assigned to floor pens within 8 light-tight rooms (15 pen replicates per F × S for 30 and 250 Hz; 18 pen replicates per F × S for 90 Hz). At 16 wk, pullets were transferred to conventional layer cages, with no flicker treatment applied. Pullet data collected included BW, feed disappearance, flock uniformity, and overall mortality. Hen data collected included BW, feed intake (feed efficiency calculated), mortality, egg production, and egg quality. Data were analyzed using Proc Mixed (SAS 9.4) and differences were considered significant when P ≤ 0.05. Frequency did not affect pullet uniformity or feed disappearance (0-8 wk and 0-16 wk). Pullets reared under 30 Hz had higher mortality (caused by "other") than those reared under 250 Hz. Lohmann Brown-Lite pullets reared under 30 Hz had the highest feed disappearance. Overall mortality was higher for LW pullets reared under 30 Hz compared to LB reared under 30 Hz or 250 Hz. Lohmann Brown-Lite hens reared under 30 Hz were heavier at the beginning of the hen phase (17 wk), however differences related to F were not seen at 40 or 48 wk. Hen day production (%) was higher for hens reared under 30 compared to 90 Hz (P = 0.03), however no other egg parameters were affected by F. Hen feed efficiency and mortality were unaffected by F. These results indicate minor effects of F, during either the pullet or hen phases. The data also suggest that S (LW vs. LB) may affect response to F.

Prediction of adeno-associated virus neutralizing antibody activity for clinical application.

Patients with neutralizing antibodies (Nabs) against adeno-associated virus (AAV) are usually excluded from the treatment with AAV vectors. To develop a standard assay for detecting Nab inhibition activity, we systematically studied current AAV Nab assays in vitro and in vivo. Several factors were found that influence the Nab titers based on the in vitro assay, including sera volume, AAV dose per cell, cell number and choice of transgenes. When the Nab titer assay was performed in vivo via intramuscular (IM) or systemic administration, a fourfold increase in sensitivity for measurement of Nab titers was observed compared with an identical in vitro test. To better mimic the clinical setting, after passively transferring human Nabs into mice, blood was collected before systemic injection of AAV vector and used for Nab titer analysis in vitro or via IM injection. The results showed that AAV delivered via IM injection had a similar inhibition pattern to systemic administration. These studies indicate critical parameters necessary for optimizing Nab sensitivity and that an in vivo Nab assay is more sensitive than an in vitro assay for inclusion/exclusion criteria. The variables identified by this study may explain some of the compounding clinical data seen to date with respect to efficiency of AAV transduction in various phase I clinical trials.

Efficient transduction of vascular smooth muscle cells with a translational AAV2.5 vector: a new perspective for in-stent restenosis gene therapy.

Coronary artery disease represents the leading cause of mortality in the developed world. Percutaneous coronary intervention involving stent placement remains disadvantaged by restenosis or thrombosis. Vascular gene therapy-based methods may be approached, but lack a vascular gene delivery vector. We report a safe and efficient long-term transduction of rat carotid vessels after balloon injury intervention with a translational optimized AAV2.5 vector. Compared with other known adeno-associated virus (AAV) serotypes, AAV2.5 demonstrated the highest transduction efficiency of human coronary artery vascular smooth muscle cells (VSMCs) in vitro. Local delivery of AAV2.5-driven transgenes in injured carotid arteries resulted in transduction as soon as day 2 after surgery and persisted for at least 30 days. In contrast to adenovirus 5 vector, inflammation was not detected in AAV2.5-transduced vessels. The functional effects of AAV2.5-mediated gene transfer on neointimal thickening were assessed using the sarco/endoplasmic reticulum Ca(2+) ATPase isoform 2a (SERCA2a) human gene, known to inhibit VSMC proliferation. At 30 days, human SERCA2a messenger RNA was detected in transduced arteries. Morphometric analysis revealed a significant decrease in neointimal hyperplasia in AAV2.5-SERCA2a-transduced arteries: 28.36±11.30 (n=8) vs 77.96±24.60 (n=10) µm(2), in AAV2.5-green fluorescent protein-infected, P<0.05. In conclusion, AAV2.5 vector can be considered as a promising safe and effective vector for vascular gene therapy.

Bladder outflow problems in females.

This is a prospective study that included 188 females who had bladder outflow problems, either due to urethral stricture disease or failure to empty the bladder. The data was taken from a computer database and included patient data since 11 years, who were on follow-up for their conditions. The women ranged from 23 to 97 years old, with a median value of 59 years. The aim of the study was to assess the bladder outflow problems in females and their treatment, which included either cystoscopy alone or in combination with urethral dilatation followed by long-term clean intermittent self catheterisation or dilatation (CISC/ISD). Out of the 188 patients, there were 135 patients who had problems with emptying the bladder due to various conditions. These females did well withCISC, which is an accepted treatment modality in this group of patients. They also used catheters of a smaller size as compared to patients using ISD for stricture disease. Thirty-eight patients among the total number had urethral stricture disease, whereas 15 patients presented with both conditions. Among the patients having treatment for stricture disease, the majority needed urethral dilatation repeated during follow-up. However, none of them needed any reconstructive surgery during that period of time. Most of the patients were comfortable doing ISD, in spite of using larger catheters comparatively. The catheters most used were the Lofric catheters. It was also seen that the majority of patients were satisfied doing CISC/ISD.

Smoking prevalence and factors associated with smoking status among Vietnamese in California.

INTRODUCTION: Vietnamese American men have smoking prevalence rates higher than the general population. We analyzed Vietnamese American smoking behavior by demographic and health-related factors, including some specific to Vietnamese, in the largest tobacco-specific survey yet targeting the Vietnamese population. METHODS: Using a statewide surname probability sample and computer-assisted telephone interviewing, we surveyed 1,101 Vietnamese men and 1,078 Vietnamese women in California (63.5% participation among successfully contacted eligible individuals) in 2007-2008. We conducted multivariate regression models to analyze the association between Vietnamese male smoking status and demographic and health-related factors. RESULTS: Among women, <1% were current smokers and <2% were former smokers. Among men, 25% were current and 24% were former smokers. Regression models for Vietnamese men delineated factors associated with both current and former smoking (vs. never smoking): being married, being employed, having lower educational attainment, and consuming alcohol. Other factors associated with current smoking (vs. never smoking) were having no health insurance, having seen a Vietnamese doctor or no doctor visit in the past year, having Vietnamese military or Vietnamese reeducation camp experience, having less knowledge about the harms of smoking, and reporting higher depression symptoms. Increasing age and not being Buddhist were associated with former (vs. never) smoking. DISCUSSION: Smoking patterns of Vietnamese women and Vietnamese men are significantly different from the general California population. Tobacco control efforts targeting Vietnamese men should include community outreach since current smokers have low health care access, utilization, and knowledge.

Intermittent self dilatation--still a viable option for treatment of urethral stricture disease.

The study was conducted to find out if intermittent self dilatation (ISD) is still a viable option for treatment of urethral strictures in patients who had undergone either urethral dilatation or urethrotomy prior to ISD. The study included 310 male patients with the age range from 17 to 93 years old from January 1996 to March 2007. The data was gathered from the computer data base kept by the urology unit in the hospital. A questionnaire was used to evaluate their tolerance towards the procedure and quality of life. Of the total number of patients 262 replied to the questionnaire. The mean follow up was 57.68 months with 67.7% continuing with the procedure. The procedure was well tolerated by 84.1% of patients and 79.6% had no technical difficulty. The most number of patients carrying out this procedure were in the 6th and 7th decade of life. There was a recurrence rate of urethral strictures of 16.9%. It is seen that even today ISD following an endoscopic procedure is a viable treatment option. The patients seem to tolerate this procedure well and it is an option in the elderly patient group who may not be fit or willing to undergo reconstructive surgery or urethroplasty.

Anti-cyclooxygenase effects of lipid extracts from the New Zealand green-lipped mussel, Perna canaliculus.

Total lipid extracts of P. canaliculus (a bivalve marine mollusc native to New Zealand, commonly called the green-lipped mussel) and Mytilus edulis (commonly called the common blue mussel) moderately inhibited ovine COX-1 and COX-2 pure enzymes in vitro. The inhibition was increased after the mussel extracts were saponified by KOH hydrolysis. Protease- and protease-lipase-hydrolysed lipid extracts of P. canaliculus exhibited similarly strong COX inhibition as the KOH-hydrolysed extract. Lyprinol(R) (a commercial extract from P. canaliculus) also exhibited strong inhibition of both COX isoforms, an effect that was increased 10-fold upon subsequent hydrolysis. In contrast, fish oil was not as anti-COX active as Lyprinol. The Lyprinol free fatty acid fraction, and to a lesser extent the Lyprinol triglyceride fraction, were the only lipid classes of Lyprinol to exhibit strong inhibition of the COX isoforms. The purified PUFA extracts were all bioactive, potently inhibiting COX-1 and COX-2. Incubation of Lyprinol in the absence of exogenous arachidonic acid (AA) showed the appearance of alternate prostaglandin metabolites, confirming Lyprinol PUFA as a competitive substrate inhibitor of AA metabolism.

Natural history of Canavan disease revealed by proton magnetic resonance spectroscopy (1H-MRS) and diffusion-weighted MRI.

Canavan disease is a childhood leukodystrophy caused by mutations in the gene for human aspartoacylase ( ASPA), which leads to an abnormal accumulation of the substrate molecule N-acetyl-aspartate (NAA) in the brain. This study was designed to model the natural history of Canavan disease using MRI and proton magnetic resonance spectroscopy ( (1)H-MRS). NAA and various indices of brain structure (morphology, quantitative T1, fractional anisotropy, apparent diffusion coefficient) were measured in white and gray matter regions during the progression of Canavan disease. A mixed-effects statistical model was used to fit all outcome measures. Longitudinal data from 28 Canavan patients were directly compared in each brain region with reference data obtained from normal, age-matched pediatric subjects. The resultant model can be used to non-invasively monitor the natural history of Canavan disease or related leukodystrophies in future studies involving drug, gene therapy, or stem cell treatments.

Immune responses to AAV in a phase I study for Canavan disease.

BACKGROUND: Canavan disease is a rare leukodystrophy with no current treatment. rAAV-ASPA has been developed for gene delivery to the central nervous system (CNS) for Canavan disease. This study represents the first use of a viral vector in an attempt to ameliorate a neurodegenerative disorder. METHODS: Subjects received intracranial infusions via six cranial burr holes. Adeno-associated virus, serotype 2 (AAV2), mediated intraparenchymal delivery of the human aspartoacylase cDNA at a maximum dose of 1 x 10(12) vector genomes per subject. The immune response and safety profiles were monitored in the follow-up of ten subjects. RESULTS: Following rAAV2 administration, we found no evidence of AAV2 neutralizing antibody titers in serum for the majority of subjects tested (7/10). In a subset (3/10) of subjects, low to moderately high levels of AAV2 neutralizing antibody with respect to baseline were detected. In all subjects, there were minimal systemic signs of inflammation or immune stimulation. In subjects with catheter access to the brain lateral ventricle, cerebrospinal fluid was examined and there was a complete absence of neutralizing antibody titers with no overt signs of brain inflammation. CONCLUSIONS: rAAV2 vector administration to the human CNS appears well tolerated. The low levels of immune response to AAV2 detected in 3/10 subjects in this study suggest at this dose and with intraparenchymal administration this approach is relatively safe. Long-term monitoring of subjects and expansion to phase II/III will be necessary in order to make definitive statements on safety and efficacy.

Effects of AAV-2-mediated aspartoacylase gene transfer in the tremor rat model of Canavan disease.

The tremor rat is a spontaneous epilepsy model with a seizure phenotype caused by a deletion in the aspartoacylase (ASPA) gene. The absence of ASPA expression in these animals results in undetectable levels of enzyme activity and the accumulation of the substrate N-acetyl-aspartate (NAA) in brain, leading to generalized myelin vacuolation and severe motor and cognitive impairment. In support of human gene therapy for CD, recombinant adeno-associated viral vector (AAV-2) expressing ASPA was stereotactically delivered to the tremor rat brain and effects on the mutant phenotype were measured. AAV-ASPA gene transfer resulted in elevated aspartoacylase bioactivity compared to untreated mutant animals and elicited a significant decrease in the pathologically elevated whole-brain NAA levels. Assessment of motor function via quantitative rotorod testing demonstrated that rats injected with AAV-ASPA significantly improved on tests of balance and coordinated locomotion compared to animals receiving control vectors. This study provides evidence that AAV-2-mediated aspartoacylase gene transfer to the brain improves biochemical and behavioral deficits in tremor rat mutants (tm/tm) and supports the rationale of human gene transfer for Canavan disease.

Do house officers learn from their mistakes?

Mistakes are inevitable in medicine. To learn how medical mistakes relate to subsequent changes in practice, we surveyed 254 internal medicine house officers. One hundred and fourteen house officers (45%) completed an anonymous questionnaire describing their most significant mistake and their response to it. Mistakes included errors in diagnosis (33%), prescribing (29%), evaluation (21%), and communication (5%) and procedural complications (11%). Patients had serious adverse outcomes in 90% of the cases, including death in 31% of cases. Only 54% of house officers discussed the mistake with their attending physicians, and only 24% told the patients or families. House officers who accepted responsibility for the mistake and discussed it were more likely to report constructive changes in practice. Residents were less likely to make constructive changes if they attributed the mistake to job overload. They were more likely to report defensive changes if they felt the institution was judgmental. Decreasing the work load and closer supervision may help prevent mistakes. To promote learning, faculty should encourage house officers to accept responsibility and to discuss their mistakes.

Viral-based gene transfer to the mammalian CNS for functional genomic studies.

A fundamental problem in neuroscience has been the creation of suitable in vivo model systems to study basic neurological phenomena and pathology of the central nervous system (CNS). Somatic cell genetic engineering with viral vectors provides a versatile tool to model normal brain physiology and a variety of neurological diseases.