Military Working Dogs Operating in Afghanistan Theater: Comparison between Pre-and Post-Mission Blood Analyses to Monitor Physical Fitness and Training

The intergovernmental organization known as the United Nations (UN) was born “to maintain international peace and security” through different operations and tasks, including “mine action” and “explosive detection”. Explosives are the most frequent cause of injuries in military personnel and an enormous danger for civilians. The role of explosive detection dogs (EDDs) and mine detection dogs has gained great consideration over time, leading to their intense use in military operations. Literature regarding working injuries reported by EDDs during missions is limited. The aim of the present study is to investigate the hematological changes that occurred between pre-and post-mission blood analyses in military working dogs deployed to Afghanistan in order to evaluate signs of health problems or physical adjustments. Examining the clinical records, only three dogs reported a medical issue, one with gastric dilatation-volvulus (GDV), and two with lameness episodes. Lack of health issues occurring during the missions was reflected by the absence of significant differences between pre-and post-mission blood analyses. Blood results were also examined by dividing the EDDs into groups considering age at departure, sex, breed and mission length. A few categories demonstrated significant changes in some parameters; however, the mean values were always included in the ranges of normality, indicating that their physical fitness and training were adequate for the required tasks

Bologna Healing Stifle Injury Index: A Comparison of Three Surgical Techniques for the Treatment of Cranial Cruciate Ligament Rupture in Dogs

The aim of this retrospective study was to test the efficacy of the Bologna Healing Stifle Injury Index (BHSII) in assessing the medium-term outcomes of dogs treated for cranial cruciate ligament rupture. This tool can be used for comparison across surgical interventions. The study population included 53 dogs with unilateral cranial cruciate ligament rupture treated using either Paatsama, Tight-Rope or tibial tuberosity advancement techniques, and 20 orthopedically sound dogs for comparative purposes. The BHSII was utilized for all the treated dogs at the time of surgery, and 1, 3, and 6 months postoperatively, while it was utilized twice in the control group. Although all the techniques achieved a successful outcome at the end of the evaluation, the application of the BHSII permitted differentiating results at each time point and stimulating discussion regarding the rapidity and degree of the healing process for each technique. It also pointed out some incongruities between the owner's and the clinician's assessment of the process. These achievements demonstrated that the BHSII should be considered by the research and clinical communities as an effective and easy tool which can be used as a repeatable and standardized method of comparison of the progress at different time points toward a final good outcome in dogs treated for cranial cruciate ligament rupture

Genetic construction, properties and application of a green fluorescent protein-tagged ciliary neurotrophic factor

The in vitro and in vivo actions of ciliary neurotrophic factor (CNTF) suggest that endogenous CNTF plays a role in nervous system development and maintenance. CNTF produces most, possibly all, of its effects by binding to a protein referred to as CNTF receptor alpha (CNTFRalpha). Information on CNTFRalpha tissue expression and dynamics would be advanced by the availability of reagents suitable for studying the subcellular localization and trafficking of CNTFRalpha. This paper describes the genetic construction, synthesis, purification and properties of a chimeric protein in which a highly fluorescent form of the green fluorescent protein (GFP) has been fused to human CNTF. The fusion protein, termed GFP-CNTF, was expressed in Escherichia coli. Histidine tagging of GFP-CNTF permitted ready purification by means of immobilized Ni(II) chromatography. Under non-reducing conditions GFP-CNTF migrated on SDS-PAGE with an apparent molecular mass of 50 kDa, although under reducing conditions it behaved electrophoretically as a 67 kDa species. Despite these discrepancies, the molecular mass of GFP-CNTF determined by mass spectrometry (54755) agreed well with its deduced relative molecular mass of 54536. Importantly, the absorbance profile of the GFP chromophore in GFP-CNTF was not modified by the presence of the CNTF domain. Moreover, the fluorescence emission spectrum of GFP-CNTF overlapped that of GFP, showing neither a change in absorbance shift nor a difference in the fluorescence quantum yield. Circular dichroism spectroscopy confirmed that the CNTF and GFP domains of GFP-CNTF folded independently of each other. GFP-tagged CNTF was equipotent to human CNTF in supporting the survival of cultured embryonic chicken sensory and ciliary ganglion neurons. GFP-CNTF, but not GFP, bound to immobilized CNTFRalpha and was displaced by an excess of human CNTF. GFP-CNTF specifically labeled the Purkinje cell layer in cerebellar slices from adult rat. This report is the first to describe a GFP chimera with a neurotrophic factor as the fusion partner. GFP-CNTF should provide a valuable tool for elucidating the role of CNTFRalpha in nervous system function

Identification, sequencing and mutagenesis of the gene for a D-carbamoylase from Agrobacterium radiobacter

A clone positive for D-carbamoylase activity (2.7 kb HindIII-BamHI DNA fragment) was obtained by screening a genomic library of Agrobacterium radiobacter in Escherichia coli. This DNA fragment contains an open reading frame of 912 bp which is predicted to encode a peptide of 304 amino acids with a calculated molecular mass of 34247 Da. The D-carbamoylase gene, named cauA, was placed under the control of T7 RNA-dependent promoter and expressed in E. coli BL21(DE3). After induction with isopropyl-thio-beta-D-galactopyranoside, the synthesis of D-carbamoylase in E. coli reached about 40% of the total protein. The expressed protein was shown to possess a molecular mass, on SDS-PAGE, of 36 kDa and showed an enhanced stability with respect to that of the wild-type enzyme derived from A. radiobacter. Site-directed mutagenesis experiments allowed us to establish that a Pro14-->Leu14 exchange leads to an inactive enzyme species, while a Cys279-->Ser279 exchange did not impair the functional properties of the enzyme