Melanocortin peptide therapeutics: historical milestones, clinical studies and commercialization

The melanocortins (MCs) are a family of multifunctional peptidergic hormones. Several superpotent, prolonged acting, enzymatically resistant, MC analogs have been designed and synthesized to help clarify the nature and role of MCs and their receptors (MCRs) in physiological functions. Two of these analogs, a linear peptide, melanotan I, and a cyclic truncated peptide, melanotan II (MTI and MTII, respectively) have been patented and tested clinically for studies on tanning of the skin (MTI) and for diagnosis and treatment of male erectile dysfunction (MTII). A new MTII analog, PT-141 (Palatin Technologies) has initial phase I/II trials and is scheduled to enter pivotal stage III clinical trials leading to commercialization. MTI may provide a therapeutic tan with the potential to lower the risk of skin cancer. PT-141 may improve sexual functionality in both males and females.     

Absence of strong strain effects in behavioral analyses of Shank3-deficient mice

Haploinsufficiency of SHANK3, caused by chromosomal abnormalities or mutations that disrupt one copy of the gene, leads to a neurodevelopmental syndrome called Phelan-McDermid syndrome, symptoms of which can include absent or delayed speech, intellectual disability, neurological changes and autism spectrum disorders. The SHANK3 protein forms a key structural part of the post-synaptic density. We previously generated and characterized mice with a targeted disruption of Shank3 in which exons coding for the ankyrin-repeat domain were deleted and expression of full-length Shank3 was disrupted. We documented specific deficits in synaptic function and plasticity, along with reduced reciprocal social interactions, in Shank3 heterozygous mice. Changes in phenotype owing to a mutation at a single locus are quite frequently modulated by other loci, most dramatically when the entire genetic background is changed. In mice, each strain of laboratory mouse represents a distinct genetic background and alterations in phenotype owing to gene knockout or transgenesis are frequently different across strains, which can lead to the identification of important modifier loci. We have investigated the effect of genetic background on phenotypes of Shank3 heterozygous, knockout and wild-type mice, using C57BL/6, 129SVE and FVB/Ntac strain backgrounds. We focused on observable behaviors with the goal of carrying out subsequent analyses to identify modifier loci. Surprisingly, there were very modest strain effects over a large battery of analyses. These results indicate that behavioral phenotypes associated with Shank3 haploinsufficiency are largely strain-independent.KEY WORDS: Shank3, Phelan-McDermid syndrome, Autism spectrum disorders, 22q13, Mouse strain, Genetic modifier, Behavior     

Impacts of colonial waterbirds on vegetation and potential restoration of island habitats

Colonial waterbirds have impacted forested island ecosystems throughout their breeding range, changing vegetation, and soil characteristics and bird communities. Our objectives were to (1) determine effects of three levels of colonial waterbird exclusion on overall vegetation diversity and growth, and survival of a candidate restoration species (black elderberry; Sambucus nigra canadensis); (2) investigate effects of different planting techniques on survival and growth of black elderberry; and (3) determine effects of waterbird colonization on soil chemistry. In 2012, we investigated effects of three levels of waterbird exclusion (none control plots [CON]; partial, which excluded waterbirds larger than gulls [PEX]; and full which excluded all waterbirds [FEX]) on bird use, existing vegetation growth and diversity, and survival of planted black elderberry on three islands in Door County, WI, Lake Michigan. In 2013, we evaluated survival of black elderberry established with four planting treatments within three waterbird exclusion treatments on two islands in 2013. We also compared soil chemistry characteristics between islands with and without nesting waterbirds for 2 years. Overall plant growth was greater in exclosures, but elderberry survival was similar among treatments. Soil replacement and weed suppression planting treatments did not affect survival, but generally increased overall elderberry biomass. Soil from nesting islands was more acidic and had greater nutrient concentrations than reference islands. Exclusion or removal of colonial nesting waterbirds from islands may improve overall vegetation growth, but successful restoration of woody vegetation may require significant soil manipulation and planting.     

Piscivorous Bird Use of Aquaculture and Natural Water Bodies in Mississippi

Double-crested cormorants (Phalacrocorax auritus) and great egrets (Ardea alba) have an extensive history of human-wildlife conflict with the aquaculture industry of western Mississippi, USA, due to their depredation of cultured catfish (Ictalurus spp.). Although aquaculture is abundant, western Mississippi also contains naturally occurring water bodies that offer alternative forage opportunities to these species. How cormorants or egrets distribute themselves among these 2 foraging options is unknown, but it has been generally assumed each species uses aquaculture disproportionately more because of the high density of available prey. To test this assumption, we surveyed these species on aquaculture and naturally occurring water bodies using aerial surveys from October through April of 2015–2016, 2016–2017, and 2017–2018. We modeled the proportion of each species on aquaculture as a function of year, date, and weather-related variables using quasi-binomial generalized linear models. Egrets used aquaculture consistently more than what was proportionally available to them and use was not influenced by any of the variables we measured. Proportional use of aquaculture by cormorants was lowest during October through January but steadily increased through April, indicating a distribution shift toward aquaculture in the months immediately prior to their migration. The highest proportional use of aquaculture by cormorants occurred in 2016, a year when lethal control measures were not allowed against cormorants. Conversely, the least proportion of cormorants on aquaculture was in 2015 when cormorants could be lethally controlled under authority of an Aquaculture Depredation Order. This trend highlights the potential influence of changes in mortality risk, caused by changes in policy regarding lethal take of cormorants, on cormorant distribution between foraging options. © 2020 The Wildlife Society.     

Fura-2 handling in a polarized epithelial barrier: the toad urinary bladder.

Toad bladders sacs were placed inside quartz cuvettes. When fura-2 AM was added to the mucosal compartment, low temperature (4 degrees C) almost completely blocked the transepithelial transfer of fluorescence observed at 20 degrees C (20 degrees C = 371 +/- 56, 4 degrees C = 29 +/- 29 fluorescence intensity in arbitrary units (FIAU), excitation at 340 nm, emission at 510 nm). Simultaneously, fluorescence accumulation inside the tissue was significantly higher (20 degrees C = 25 +/- 5, 4 degrees C = 91 +/- 24% increase on basal levels (%IBL)). When fura-2 AM was added to the serosal side, low temperature also reduced the serosal to mucosal transfer (20 degrees C = 149 +/- 36, 4 degrees C = 61 +/- 35 FIAU). Nevertheless, in this situation tissue accumulation, that was significantly higher that the one observed when fura-2 AM was added to the mucosal side, was reduced at low temperature (20 degrees C = 300 +/- 30, 4 degrees C = 48 +/- 7 %IBL). Spectral analysis of the mucosal and serosal compartments indicated that free fura-2 was transferred from the intracellular to the serosal compartment, but not to the mucosal one. These results indicate that fura-2 appears as a useful tool to evaluate the cellular distribution and traffic of polycyclic charged and non-charged molecules.     

Simulation of fretting wear at orthopaedic implant interfaces

Osteolysis due to wear debris is a primary cause of failure of total joint replacements. Although debris produced by the joint articulating surfaces has been studied and simulated extensively, fretting wear debris, produced at nonarticulating surfaces, has not received adequate attention. We developed a three-station fretting wear simulator to reproduce in vivo motion and stresses at the interfaces of total joint replacements. The simulator is based on the beam bending theory and is capable of producing cyclic displacement from 3 to 1000 microns, under varying magnitudes of contact stresses. The simulator offers three potential advantages over previous studies: The ability to control the displacement by load, the ability to produce very small displacements, and dynamic normal loads as opposed to static. A pilot study was designed to test the functionality of the simulator, and verify that calculated displacements and loads produced the predicted differences between two commonly used porous ingrowth titanium alloy surfaces fretting against cortical bone. After 1.5 million cycles, the simulator functioned as designed, producing greater wear of bone against the rougher plasma-sprayed surface compared to the fiber-mesh surface, as predicted. A novel pin-on-disk apparatus for simulating fretting wear at orthopaedic implant interfaces due to micromotion is introduced. The test parameters measured with the fretting wear simulator were as predicted by design calculations, and were sufficient to measure differences in the height and weight of cortical bone pins rubbing against two porous ingrowth surfaces, plasma-sprayed titanium and titanium fiber mesh.