Cloning of a membrane protein that induces a slow voltage-gated potassium current

A rat kidney messenger RNA that induces a slowly activating, voltage-dependent potassium current on its expression in Xenopus oocytes was identified by combining molecular cloning with an electrophysiological assay. The cloned complementary DNA encodes a novel membrane protein that consists of 130 amino acids with a single putative transmembrane domain. This protein differs from the known ion channel proteins but is involved in the induction of selective permeation of potassium ions by membrane depolarization.     

Analysis of antibody response by temperature-sensitive measles vaccine strain in the cotton rat model

Measles virus (MeV) vaccine strain, AIK-C, is temperature sensitive (ts), which is thought to be associated with attenuation of virus pathogenicity. In this study, replication and antibody response were examined in cotton rats using viruses carrying different forms of the P gene, which is responsible for the ts phenotype of strain AIK-C and its parental Edmonston strain. When cotton rats were inoculated intranasally, ts viruses neither replicated in lungs, nor reproducibly generated an antibody response. When inoculated intramusculary (i.m.), however, ts strains raised an antibody titer in all animals. This response was not observed when ultraviolet-inactivated virus was used. ts virus, inoculated i.m., was recovered from cotton rat drainage lymph nodes. These results suggest that ts virus, inoculated i.m., could replicate in the cotton rat, presumably at the superficial lymph node, and induce an antibody response. Therefore, cotton rats can serve as a small-animal model for investigating immune responses to safer ts vaccine, as well as recombinant vaccine using AIK-C as a vector for protection against other infectious agents.     

Single spaghetti tagging as a high-retention marking method for Japanese common sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

Populations of sea cucumbers, including the Japanese common sea cucumber Apostichopus japonicus, have been seriously depleted worldwide due to overfishing. Mark–recapture study is an efficient means of collecting ecological data. However, the use of such a method in sea cucumbers is difficult because they lack hard tissues in the body wall. Here we tested the viability of various tagging methods on A. japonicus. First, we applied conventional tags using four different methods [single spaghetti (T-bar) tagging, double spaghetti tagging, ribbon tagging, and Atkins tagging] to ten individuals per method in aquaria for 14 days. Of the methods used, single spaghetti tagging had the highest retention rate. Then we examined the retention rate of single spaghetti tags on ten individual sea cucumbers for up to approximately 6 months in rearing conditions. The single spaghetti tagging method showed a retention rate of 100% over at least 7 days, and 50% of the tags remained embedded after 56 days. The longest duration of tag retention was 174 days, at which time the experiment was terminated. These results indicate that single spaghetti tagging is reliable for both short- and longer-term studies, making it a useful tool for ecological and conservation studies in sea cucumbers.     

Evaluation of a d-Octaarginine-linked polymer as a transfection tool for transient and stable transgene expression in human and murine cell lines

Poly(N-vinylacetamide-co-acrylic acid) coupled with d-octaarginine (VP-R8) promotes the cellular uptake of peptides/proteins in vitro; however, details of the transfection efficacy of VP-R8, such as the cell types possessing high gene transfer, are not known. Herein, we compared the ability of VP-R8 to induce the cellular uptake of plasmid DNA in mouse and human cell lines from different tissues and organs. A green fluorescent protein (GFP)-expression plasmid was used as model genetic material, and fluorescence as an indicator of uptake and plasmid-derived protein expression. Three mouse and three human cell lines were incubated with a mixture of plasmid and VP-R8, and fluorescence analysis were performed two days after transfection. To confirm stable transgene expression, we performed drug selection three days after transfection. A commercially available polymer-based DNA transfection reagent (PTR) was used as the transfection control and standard for comparing transgene expression efficiency. In the case of transient transgene expression, slight-to-moderate GFP expression was observed in all cell lines transfected with plasmid via VP-R8; however, transfection efficiency was lower than using the PTR for gene delivery. In the case of stable transgene expression, VP-R8 promoted drug-resistance acquisition more efficiently than the PTR did. Cells that developed drug resistance after VP-R8-mediated gene transfection expressed GFP more efficiently than cells that developed drug resistance after transfection with the PTR. Thus, VP-R8 shows potential as an in vitro or ex vivo nonviral transfection tool for generating cell lines with stable transgene expression.     

Surgical outcomes in dogs with tracheal collapse treated with a novel cross-and-hook braided endoluminal stent

BackgroundStenting is an effective treatment option for tracheal collapse in dogs. Cross-braided tracheal stents are currently the norm in veterinary medicine, but cross-and-hook braided stents have recently been adopted in human medicine. We examined whether stents manufactured using this novel braiding technique provided additional advantages for the treatment of tracheal collapse in dogs.ObjectivesTo evaluate the outcomes of cross-and-hook braided stent implantation in the treatment of tracheal collapse in dogs.MethodsThe medical records of 22 client-owned dogs that underwent luminal placement of cross-and-hook braided Fauna Stents for the treatment of tracheal collapse between January 2018 and July 2021 were examined and data on canine signalment, clinical signs, diagnostic test results, surgical outcomes, and postoperative complications were retrieved and analyzed statistically.ResultsTwenty-six stents were surgically implanted, with 20 dogs (90.9%) receiving one stent and the remaining two (9.1%) receiving two or more stents. All dogs survived the procedure. The median survival time at a median follow-up of 990 days was 879 days. At the final follow-up examination, loss or mild improvement of cough was observed in all dogs.ConclusionsCompared with conventional lumen stents, the cross-and-hook braided Fauna Stent offered a higher survival rate and improved clinical symptoms in all patients. The results of this study suggest that the Fauna Stent may be a promising treatment option for dogs with tracheal collapse.     

Clip ligation for treatment of patent ductus arteriosus occlusion in three cats

Patent ductus arteriosus (PDA) is a rare congenital cardiovascular anomaly in cats. Due to their small body, intercostal thoracotomy is the most common option to close the PDA. However, few reports detail the surgical technique for ligating PDA in kittens. In this case report, three cats weighing 1.4 kg, 1.2 kg, and 2.9 kg were diagnosed PDA. Clip ligation via left fourth intercostal thoracotomy was performed and the cats were successfully treated. Postoperative echocardiography showed no residual flow in any of the cases. This case report highlights clip occlusion for small cats with PDA could be safe and effective.