The effect of oral corticosterone, prolactin and prolactin deprivation on weight gain and locomotor function in neonatal rats

In order to simulate the elevated corticosteroid and prolactin levels that offspring of stressed mothers may be subjected during breast feeding, rat pups were treated daily with oral corticosterone (200ng/ml milk intake) or prolactin (140ng/ml milk intake) from the 2nd to the 151h postnatal day. To investigate the potential influence of reduced prolactin intake, the mothers were either treated with bromocriptine (2ug—12ug/rat/day) or 1% ethanol (vehicle). The rat pups were subjectedto swim tests from their 8th postnatal day to examine their neuromuscular development. Results from swim tests showed latency in development in the prolactin, corticosterone and prolactin deprived/cortieosterone administered groups, compared with the controls. There was decreased daily weight gains in the treatment groups compared to the control. This study demonstrates that increased prolactin and corticosterone and decreased prolactin combined with elevated enrticoisterone levels to which suckling neonates were exposed to, had a significant negative effect on their neuromuscular adaptive mechanism involved in the normal development of the locomotor system

Teaching a culture of care: Why it matters

The use of animal in biomedical research remains a critical compromise. Research and higher education institutions play a major role in educating on the use of animal and such training is expected to translate into the development of a culture of care practice across all staff working with animals. But nurturing a “culture of care” and impacting in professional attitudes in the field of animal research remains challenging due to its social, ethical and different institutional frameworks. From an educational perspective, current practice remains challenged by the need for better integration of inter-cultural perceptions on animal welfare, supported by more cross disciplinary integration in educational curriculum including the relevance of the 3Rs principles and promoting reflective practice strategies.  Institutional support is crucial to provide a safe, and supportive framework to promote such caring ethos. Our aim is to discuss practical actions to implement and assess culture of care, highlighting its direct impact on the professional integrity of staff which is directly linked to research and education excellence. Seeking a global welfare for all the beings involved and supporting individual and team reflective practice will provide better tools to guarantee the best care of the animals

Assessment of F/HN-Pseudotyped Lentivirus as a Clinically Relevant Vector for Lung Gene Therapy

RATIONALE: Ongoing efforts to improve pulmonary gene transfer thereby enabling gene therapy for the treatment of lung diseases, such as cystic fibrosis (CF), has led to the assessment of a lentiviral vector (simian immunodeficiency virus [SIV]) pseudotyped with the Sendai virus envelope proteins F and HN. OBJECTIVES: To place this vector onto a translational pathway to the clinic by addressing some key milestones that have to be achieved. METHODS: F/HN-SIV transduction efficiency, duration of expression, and toxicity were assessed in mice. In addition, F/HN-SIV was assessed in differentiated human air-liquid interface cultures, primary human nasal epithelial cells, and human and sheep lung slices. MEASUREMENTS AND MAIN RESULTS: A single dose produces lung expression for the lifetime of the mouse (~2 yr). Only brief contact time is needed to achieve transduction. Repeated daily administration leads to a dose-related increase in gene expression. Repeated monthly administration to mouse lower airways is feasible without loss of gene expression. There is no evidence of chronic toxicity during a 2-year study period. F/HN-SIV leads to persistent gene expression in human differentiated airway cultures and human lung slices and transduces freshly obtained primary human airway epithelial cells. CONCLUSIONS: The data support F/HN-pseudotyped SIV as a promising vector for pulmonary gene therapy for several diseases including CF. We are now undertaking the necessary refinements to progress this vector into clinical trials

Chapter 12: Bioluminescent Monitoring In Vivo Colonisation and Clearance Dynamics by LightEmitting Bacteria, Bioluminescence ,Methods in Molecular Biology. Human press, a part of Springer Science and Business Media LCC

Abstract Bioluminescence is an excellent reporter system for analysing bacterial colonization and clearance dynamics in vivo. Many bacterial species have been rendered bioluminescent, allowing the sensitive detection of bacterial burden and metabolic activity in real-time and in situ in living animals. In this chapter we describe the protocols for characterizing in vivo infection models using bioluminescent bacteria: from real-time imaging in living animals by bioluminescence imaging (BLI) to ex vivo BLI of harvested organs and tissues and, finally, to quantification of bacterial numbers in organ and tissue homogenates by luminometry and viable counts. While the lux operon from Photorhabdus luminescens is ideally suited for use in such models, there may be times when alternative luciferases, such as those from the firefly (luc) or marine copepods (Gluc), may be more appropriate. Here we describe the protocols required to monitor colonization and clearance dynamics using bioluminescent bacteria that are lux-, luc-, or Gluc-positive

Use of biophotonic imaging as a training aid for administration of substances in laboratory rodents.

Dosing of experimental animals and the removal of blood are two of the most frequent procedures performed in biomedical research using live animals. Despite the apparently simple nature of these procedures, they can, if not correctly carried out, have significant effects on the welfare of the animals and the scientific value of the results. There are several methods by which research staff may obtain training in the administration of substances. These include practical demonstrations during teaching courses; observation of techniques; videos and educational computer programs and practising on recently killed animal cadavers or plastic animal models. Each method has its own advantages and disadvantages. A common factor encountered during training is the difficulty in assessing competency. This paper reports a pilot study on the use of bioluminescent imaging technology to assess competency in the administration of substances to rodents. Bioluminescence was rapidly detected after dosing of animals with a bioluminescent substance. However, living animals were required for a signal to be generated. The data presented suggest that this technology is ideal for use as a teaching aid and may also prove valuable in assessing the effectiveness of 'complex' and novel administration routes in 'realtime'

Microbial hazards during harvesting and processing at an outdoor United States cannabis farm

<p>Cannabis cultivation is an emerging industry within the United States. Organic dust derived in part from naturally occurring microorganisms is known to cause byssinosis in the hemp industry. In this pilot study, bacteria and fungi encountered by workers at an outdoor cannabis farm that utilized organic practices were elucidated by 16 S ribosomal RNA (rRNA) and Internal Transcribed Spacer (ITS) region sequencing, respectively. Area (n = 14) and personal air samples (n = 12) were collected during harvesting and processing activities. 16 S rRNA and ITS regions of extracted bacterial and fungal genomic DNA were amplified and sequenced using Sanger sequencing. Bacterial sequencing resolved 1,077 sequences that were clustered into 639 operational taxonomic units (OTUs) and predominantly placed in the phylum, Actinobacteria (46%). Personal air samples revealed higher bacterial and Actinobacteria diversity compared to outdoor area samples collected within the facility (<i>p</i> < 0.05). A high degree of dissimilarity between bacteria was identified within and between samples. Fungal sequences (n = 985) were identified and predominantly clustered in the phylum Ascomycota (53%). Of the 216 fungal OTUs elucidated, the cannabis plant pathogenic species, <i>Botrytis cinerea</i>, was the most prevalent and accounted for 34% of all fungal sequences. The relative abundance of <i>B. cinerea</i> was highest in personal air samples (59%) compared to area samples collected in the drying room (19%), greenhouse (18%), and outdoor environment (6%). There was 49% sample similarity between fungi identified within personal air samples, but higher dissimilarity coefficients were observed within and between greenhouse, drying room, and outdoor area air samples. The results of this pilot study suggest that the cannabis farm workers are potentially exposed to Actinobacteria as well as the cannabis plant pathogen, <i>B. cinerea</i> during harvesting, bud-stripping, and hand-trimming processes.</p

Accelerated syntheses of amine-bis(phenol) ligands in polyethylene glycol or “on water” under microwave irradiation

Pure amine-bis(phenol) ligands are readily accessible in high yield, often >90%, when the Mannich condensation reactions are performed “on water” or in poly(ethyleneglycol) (PEG). Microwave-assisted synthesis dramatically reduces the time and energy required to prepare these molecules, typically from 24 h to 5 min. The approach seems to be widely applicable (7 amines and 5 phenols were tested to yield a diverse set of bis(phenol) ligands). Significant improvements in yield were observed for ligands derived from di-tert-amyl and di-tert-butyl phenols, possibly resulting from a hydrophobic effect. Single crystal X-ray diffraction data for the ligand derived from p-cresol and N,N′-dimethylethylenediamine is reported

Sex Inclusive Research Framework (SIRF): an evaluation tool to assess whether an in vivo research proposal follows the sex inclusive research philosophy.

A new, interactive framework supports the evaluation of in vivo and ex vivo research proposals from a sex inclusive research perspective delivering a traffic light classification, indicating whether a proposal is appropriate, risky, or insufficient with regards to sex inclusion. This tool is designed for use by researchers, (animal) ethical review boards, and funders to generate a rigorous and reproducible assessment of sex inclusion at the proposal level, thus helping address the embedded sex bias in preclinical research