Phosphoregulation accommodates Type III secretion and assembly of a tether of ER-Chlamydia inclusion membrane contact sites.

Membrane contact sites (MCS) are crucial for nonvesicular trafficking-based interorganelle communication. Endoplasmic reticulum (ER)-organelle tethering occurs in part through the interaction of the ER resident protein VAP with FFAT motif-containing proteins. FFAT motifs are characterized by a seven amino acidic core surrounded by acid tracks. We have previously shown that the human intracellular bacterial pathogen Chlamydia trachomatis establishes MCS between its vacuole (the inclusion) and the ER through expression of a bacterial tether, IncV, displaying molecular mimicry of eukaryotic FFAT motif cores. Here, we show that multiple layers of host cell kinase-mediated phosphorylation events govern the assembly of the IncV-VAP tethering complex and the formation of ER-Inclusion MCS. Via a C-terminal region containing three CK2 phosphorylation motifs, IncV recruits CK2 to the inclusion leading to IncV hyperphosphorylation of the noncanonical FFAT motif core and serine-rich tracts immediately upstream of IncV FFAT motif cores. Phosphorylatable serine tracts, rather than genetically encoded acidic tracts, accommodate Type III-mediated translocation of IncV to the inclusion membrane, while achieving full mimicry of FFAT motifs. Thus, regulatory components and post-translational modifications are integral to MCS biology, and intracellular pathogens such as C. trachomatis have evolved complex molecular mimicry of these eukaryotic features.

Clinical validation of a next-generation sequencing-based multi-cancer early detection "liquid biopsy" blood test in over 1,000 dogs using an independent testing set: The CANcer Detection in Dogs (CANDiD) study.

Cancer is the leading cause of death in dogs, yet there are no established screening paradigms for early detection. Liquid biopsy methods that interrogate cancer-derived genomic alterations in cell-free DNA in blood are being adopted for multi-cancer early detection in human medicine and are now available for veterinary use. The CANcer Detection in Dogs (CANDiD) study is an international, multi-center clinical study designed to validate the performance of a novel multi-cancer early detection "liquid biopsy" test developed for noninvasive detection and characterization of cancer in dogs using next-generation sequencing (NGS) of blood-derived DNA; study results are reported here. In total, 1,358 cancer-diagnosed and presumably cancer-free dogs were enrolled in the study, representing the range of breeds, weights, ages, and cancer types seen in routine clinical practice; 1,100 subjects met inclusion criteria for analysis and were used in the validation of the test. Overall, the liquid biopsy test demonstrated a 54.7% (95% CI: 49.3-60.0%) sensitivity and a 98.5% (95% CI: 97.0-99.3%) specificity. For three of the most aggressive canine cancers (lymphoma, hemangiosarcoma, osteosarcoma), the detection rate was 85.4% (95% CI: 78.4-90.9%); and for eight of the most common canine cancers (lymphoma, hemangiosarcoma, osteosarcoma, soft tissue sarcoma, mast cell tumor, mammary gland carcinoma, anal sac adenocarcinoma, malignant melanoma), the detection rate was 61.9% (95% CI: 55.3-68.1%). The test detected cancer signal in patients representing 30 distinct cancer types and provided a Cancer Signal Origin prediction for a subset of patients with hematological malignancies. Furthermore, the test accurately detected cancer signal in four presumably cancer-free subjects before the onset of clinical signs, further supporting the utility of liquid biopsy as an early detection test. Taken together, these findings demonstrate that NGS-based liquid biopsy can offer a novel option for noninvasive multi-cancer detection in dogs.

In vitro evaluation of negative pressure generated during application of negative suction volumes by use of various syringes with and without thoracostomy tubes.

OBJECTIVE: To determine the amount of negative pressure generated by syringes of various sizes with and without an attached thoracostomy tube and whether composition of thoracostomy tubes altered the negative pressure generated. SAMPLE: Syringes ranging from 1 to 60 mL and 4 thoracostomy tubes of various compositions (1 red rubber catheter, 1 polyvinyl tube, and 2 silicone tubes). PROCEDURES: A syringe or syringe with attached thoracostomy tube was connected to a pneumatic transducer. Each syringe was used to aspirate a volume of air 10 times. Negative pressure generated was measured and compared among the various syringe sizes and various thoracostomy tubes. RESULTS: The negative pressure generated decreased as size of the syringe increased for a fixed volume across syringes. Addition of a thoracostomy tube further decreased the amount of negative pressure. The red rubber catheter resulted in the least amount of negative pressure, followed by the polyvinyl tube and then the silicone tubes. There was no significant difference in negative pressure between the 2 silicone tubes. The smallest amount of negative pressure generated was -74 to -83 mm Hg. CONCLUSIONS AND CLINICAL RELEVANCE: Limited data are available on the negative pressure generated during intermittent evacuation of the thoracic cavity. For the present study, use of a syringe of ≥ 20 mL and application of 1 mL of negative suction volume resulted in in vitro pressures much more negative than the currently recommended pressure of -14.71 mm Hg for continuous suction. Additional in vitro or cadaveric studies are needed.

The use of rim excision as a treatment for canine acanthomatous ameloblastoma.

This study reviews rim excision as a treatment for canine acanthomatous ameloblastomas (CAA) in dogs with <3 mm of bone involvement. Removal of a canine tooth was involved in 47% of the cases; 33% cases involved the caudal dentition. Follow-up ranged from 3 months to 5 years. No evidence of recurrence was seen. Client satisfaction with cosmesis and the animal's ability to masticate was judged to be good. With appropriate case selection, rim excision appears to be a viable option for CAA and results in improved dental occlusion, cosmesis, and no evidence of epulis recurrence.