Larval Therapy and Larval Excretions/Secretions: A Potential Treatment for Biofilm in Chronic Wounds? A Systematic Review.

Chronic wounds present a global healthcare challenge and are increasing in prevalence, with bacterial biofilms being the primary roadblock to healing in most cases. A systematic review of the to-date knowledge on larval therapy's interaction with chronic-wound biofilm is presented here. The findings detail how larval therapy-the controlled application of necrophagous blowfly larvae-acts on biofilms produced by chronic-wound-relevant bacteria through their principle pharmacological mode of action: the secretion and excretion of biologically active substances into the wound bed. A total of 12 inclusion-criteria-meeting publications were identified following the application of a PRISMA-guided methodology for a systematic review. The findings of these publications were qualitatively analyzed to provide a summary of the prevailing understanding of larval therapy's effects on bacterial biofilm. A further review assessed the quality of the existing evidence to identify knowledge gaps and suggest ways these may be bridged. In summary, larval therapy has a seemingly unarguable ability to inhibit and degrade bacterial biofilms associated with impaired wound healing. However, further research is needed to clarify and standardize the methodological approach in this area of investigation. Such research may lead to the clinical application of larval therapy or derivative treatments for the management of chronic-wound biofilms and improve patient healing outcomes at a time when alternative therapies are desperately needed.

Efficient Synthesis for Altering Side Chain Length on Cannabinoid Molecules and Their Effects in Chemotherapy and Chemotherapeutic Induced Neuropathic Pain.

(1) Background: Recently, a number of side chain length variants for tetrahydrocannabinol and cannabidiol have been identified in cannabis; however, the precursor to these molecules would be based upon cannabigerol (CBG). Because CBG, and its side chain variants, are rapidly converted to other cannabinoids in the plant, there are typically only small amounts in plant extracts, thus prohibiting investigations related to CBG and CBG variant therapeutic effects. (2) Methods: To overcome this, we developed an efficient synthesis of corresponding resorcinol fragments using the Wittig reaction which, under acid catalyzed coupling with geraniol, produced the desired side chain variants of CBG. These compounds were then tested in an animal model of chemotherapeutic-induced neuropathic pain and to reduce colorectal cancer cell viability. (3) Results: We found that all side-chain variants were similarly capable of reducing neuropathic pain in mice at a dose of 10 mg/kg. However, the molecules with shorter side chains (i.e., CBGV and CBGB) were better at reducing colorectal cancer cell viability. (4) Conclusions: The novel synthesis method developed here will be of utility for studying other side chain derivatives of minor cannabinoids such as cannabichromene, cannabinol, and cannabielsoin.

The Importance of Evaluating Positive Welfare Characteristics and Temperament in Working Therapy Dogs.

To date, investigations of the welfare of therapy dogs have focused largely on examining physiological and behavioral measures that could indicate if the animal is experiencing stress or distress. However, this approach does not fully address the definition of welfare which is often described as existing on a continuum from negative (or stressful) to positive. With therapy dogs, it would be worth addressing if they experience positive emotional affect while working since the quality and efficacy of animal-assisted interventions for the human recipient is likely to be influenced by the animal's emotional state during the interaction. The purpose of this review is to articulate how objective measurements of the HPA axis and measurements of behavioral observations and standardized questions can be used to evaluate positive welfare in therapy dogs. A potentially relevant indicator of positive welfare is the peripheral concentration of the neurohormone oxytocin, which has been found to increase in systemic circulation within a variety of species during positive social and affiliative contexts, including during human-dog interaction. Oxytocin is also a negative-feedback regulator of the Hypothalamic-Pituitary-Adrenal (HPA) axis, which culminates with the production of the stress hormone cortisol. Cortisol is widely used as a physiological indicator to assess negative welfare states in animals, including therapy dogs. Observable behavior during interactions with humans that may convey enjoyment could provide indicators of positive welfare in dogs such as engagement in play, or human-directed affiliative behaviors including leaning against, nudging, or licking the patient. However, in assessing positive welfare, it is also critical to consider that all animal behavioral displays and physiological responses are dependent on the dog's individual (and breed) temperament. Temperament directly drives how the animal copes and responds to its current physical and social environment, including during stressful situations such as when therapy dogs interact with unfamiliar humans in novel healthcare settings. Coupled with both positive and negative physiological and behavioral welfare indicators, questionnaire data can provide further context to, and enhance interpretations of, therapy dog welfare assessment results. Overall, to date, no studies have measured all of these factors to assess therapy dog welfare.

The Role of Mitochondria-Linked Fatty-Acid Uptake-Driven Adipogenesis in Graves Orbitopathy.

CONTEXT: Depot-specific expansion of orbital adipose tissue (OAT) in Graves orbitopathy (GO; an autoimmune condition producing proptosis, visual impairment and reduced quality of life) is associated with fatty acid (FA)-uptake-driven adipogenesis in preadipocytes/fibroblasts (PFs). OBJECTIVE: This work sought a role for mitochondria in OAT adipogenesis in GO. METHODS: Confluent PFs from healthy OAT (OAT-H), OAT from GO (OAT-GO) and white adipose tissue in culture medium compared with culture medium containing a mixed hormonal cocktail as adipogenic medium (ADM), or culture-medium containing FA-supplementation, oleate:palmitate:linoleate (45:30:25%) with/without different concentration of mitochondrial biosubstrate adenosine 5'-diphosphate/guanosine 5'-diphosphate (ADP/GDP), AICAR (adenosine analogue), or inhibitor oligomycin-A for 17 days. Main outcome measures included oil-red-O staining and foci count of differentiated adipocytes for in vitro adipogenesis, flow cytometry, relative quantitative polymerase chain reaction, MTS-assay/106 cells, total cellular-ATP detection kit, and Seahorse-XFe96-Analyzer for mitochondria and oxidative-phosphorylation (OXPHOS)/glycolysis-ATP production analysis. RESULTS: During early adipogenesis before adipocyte formation (days 0, 4, and7), we observed OAT-specific cellular ATP production via mitochondrial OXPHOS in PFs both from OAT-H and OAT-GO, and substantially disrupted OXPHOS-ATP/glycolysis-ATP production in PFs from OAT-GO, for example, a 40% reduction in OXPHOS-ATP and trend-increased glycolysis-ATP production on days 4 and 7 compared with day 0, which contrasted with the stable levels in OAT-H. FA supplementation in culture-medium triggered adipogenesis in PFs both from OAT-H and OAT-GO, which was substantially enhanced by 1-mM GDP reaching 7% to 18% of ADM adipogenesis. The FA-uptake-driven adipogenesis was diminished by oligomycin-A but unaffected by treatment with ADP or AICAR. Furthermore, we observed a significant positive correlation between FA-uptake-driven adipogenesis by GDP and the ratios of OXPHOS-ATP/glycolysis-ATP through adipogenesis of PFs from OAT-GO. CONCLUSION: Our study confirmed that FA uptake can drive OAT adipogenesis and revealed a fundamental role for mitochondria-OXPHOS in GO development, which provides potential for therapeutic interventions.

Detection and Inhibition of IgE for cross-reactive carbohydrate determinants evident in an enzyme-linked immunosorbent assay for detection of allergen-specific IgE in the sera of dogs and cats.

BACKGROUND: It has been demonstrated recently that immunoglobulin (Ig)E specific for cross-reactive carbohydrate determinants (CCD) is present in the serum of allergen-sensitized dogs and cats, and that these CCD-specific antibodies might confound serological testing. HYPOTHESIS/OBJECTIVE: The objective was to document the prevalence of CCD detectable in a monoclonal cocktail-based enzyme-linked immunosorbent assay designed for the detection of allergen-specific IgE in the sera of dogs and cats, and to define a means for successful inhibition of these CCD. METHODS AND MATERIALS: The incidence of reactivity to bromelain and a commercially available inhibitor of carbohydrate-specific antibodies (RIDA-CCD) was evaluated in 100 dog sera samples before and after inhibition with RIDA-CCD and a proprietary inhibitor containing carbohydrates derived from bromelain (BROM-CCD). Subsequently, sera from 600 dogs and 600 cats were evaluated using a serum diluent with and without BROM-CCD. RESULTS: Both the RIDA-CCD and BROM-CCD inhibitors demonstrated successful reduction of CCD reactivity, although a more efficient profile of inhibition was evident with BROM-CCD. Mite reactivity in dog and cat sera was largely unaffected; however, substantial inhibition for pollen allergens (trees, grasses and weeds) was shown. After BROM-CCD inhibition, 1% of canine samples and 13% of feline samples were rendered completely negative for allergen reactivity. CONCLUSIONS AND CLINICAL IMPORTANCE: The results demonstrate that BROM-CCD is effective in reducing reactions with irrelevant carbohydrates, and that inhibition of CCD reactivity might substantially alter the outcome of the in vitro reactivity profile used for selection of allergens to be included in an immunotherapeutic regime.

Risks associated with animal-assisted intervention programs: A literature review.

The benefits of animal-assisted interventions (AAI), to utilize companion animals as an adjunctive treatment modality, is well-established and a burgeoning research field. However, few studies have evaluated the potential hazards of these programs, such as the potential for therapy animals to transfer hospital-associated pathogens between individuals and the hospital environment. Here we review the current literature on the possible risks of hospital-based AAI programs, including zoonotic pathogen transmission. We identified twenty-nine articles encompassing reviews of infection control guidelines and epidemiological studies on zoonotic pathogen prevalence in AAI. We observed substantial heterogeneity in infection control practices among hospital AAI programs. Few data confirmed pathogen transmission between therapy animals and patients. Given AAI's known benefits, we recommend that future research utilize a One Health framework to evaluate microbial dynamics among therapy animals, patients, and hospital environments. This framework may best promote safe practices to ensure the sustainability of these valuable AAI programs.

An NMR-Guided Screening Method for Selective Fragment Docking and Synthesis of a Warhead Inhibitor.

Selective hits for the glutaredoxin ortholog of Brucella melitensis are determined using STD NMR and verified by trNOE and (15)N-HSQC titration. The most promising hit, RK207, was docked into the target molecule using a scoring function to compare simulated poses to experimental data. After elucidating possible poses, the hit was further optimized into the lead compound by extension with an electrophilic acrylamide warhead. We believe that focusing on selectivity in this early stage of drug discovery will limit cross-reactivity that might occur with the human ortholog as the lead compound is optimized. Kinetics studies revealed that lead compound 5 modified with an ester group results in higher reactivity than an acrylamide control; however, after modification this compound shows little selectivity for bacterial protein versus the human ortholog. In contrast, hydrolysis of compound 5 to the acid form results in a decrease in the activity of the compound. Together these results suggest that more optimization is warranted for this simple chemical scaffold, and opens the door for discovery of drugs targeted against glutaredoxin proteins-a heretofore untapped reservoir for antibiotic agents.

PREGO (presentation of Graves' orbitopathy) study: changes in referral patterns to European Group On Graves' Orbitopathy (EUGOGO) centres over the period from 2000 to 2012.

BACKGROUND/AIMS: The epidemiology of Graves' orbitopathy (GO) may be changing. The aim of the study was to identify trends in presentation of GO to tertiary centres and initial management over time. METHODS: Prospective observational study of European Group On Graves' Orbitopathy (EUGOGO) centres. All new referrals with a diagnosis of GO over a 4-month period in 2012 were included. Clinical and demographic characteristics, referral timelines and initial decisions about management were recorded. The data were compared with a similar EUGOGO survey performed in 2000. RESULTS: The demographic characteristics of 269 patients studied in 2012 were similar to those collected in the year 2000, including smoking rates (40.0% vs 40.2%). Mild (60.5% vs 41.2%, p<0.01) and inactive GO (63.2% vs 39.9%, p<0.01) were more prevalent in 2012. The times from diagnosis of thyroid disease to being seen in EUGOGO centres (6 vs 16 months) and from first symptoms of GO (9 vs 16 months) or from diagnosis of GO (6 vs 12 months) to first consultation in EUGOGO centres were shorter in 2012 (p<0.01). The initial management plans for GO were no different except surgical treatments for patients with mild inactive disease were more frequently offered in the 2012 cohort than in 2000 (27.3% vs 17%, p<0.05), and selenium supplements were offered only in the 2012 cohort (21.2% vs 0%, p<0.01). CONCLUSIONS: These findings suggest that the clinical manifestations of patients with GO may be changing over time in Europe.

Vibration-assisted anesthesia in eyelid surgery.

PURPOSE: To investigate whether a vibrating device applied in a circular motion to the forehead reduces the pain of local anesthetic injection in upper eyelid surgery. DESIGN: Prospective, interventional, cross-over, randomized, controlled clinical trial. PARTICIPANTS: Eighty patients undergoing bilateral upper eyelid surgery under local anesthesia. INTERVENTION: A vibrating device was applied in a circular motion to participants' foreheads while local anesthetic was injected into 1 eyelid. During injection of anesthetic on the contralateral lid, the device was applied to the forehead in static fashion with the vibration switched off (placebo). The order of intervention was randomized. MAIN OUTCOME MEASURES: After both injections had been given, participants were asked to specify their pain ratings for each injection on a scale of 0 to 10, with 0 representing no pain and 10 indicating the worst pain imaginable. Participants were also asked to qualitatively compare the 2 sides. RESULTS: The mean pain scores were 3.3 for the vibration-assisted side and 4.5 for the placebo. This difference was statistically significant (P=0.0003); 73% of participants found the vibrated side to be better than the placebo, with 35% finding it a lot or quite a bit better. CONCLUSIONS: Vibration-assisted anesthesia during upper eyelid surgery has a beneficial effect that is highly statistically significant and is clinically significant in terms of patients' qualitative assessment of pain. Further research is needed to determine whether this constitutes a quantitatively clinically significant improvement in pain management. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Cataract surgery and quality of life implications.

Cataract surgery in the developed world has undergone a revolution over the last 20 years. An operation which used to require a stay in hospital and long visual rehabilitation is now a quick day-case procedure with immediate benefits. As with any surgery there is an associated morbidity, but there is now the potential to provide cataract surgery at an earlier stage of cataract maturation and save patients from a period of severe visual impairment. This article reviews the new techniques available to measure the impact that cataracts have not only on a patient's visual acuity but also their general physical health, function, cognition, and emotional well-being. New research is described that takes into account these more holistic tests and how they can be used to judge the best time to refer and operate on a patient with cataracts.