A novel placental tissue biologic, PTP-001, inhibits inflammatory and catabolic responses in vitro and prevents pain and cartilage degeneration in a rat model of osteoarthritis.

OBJECTIVE: Characterization of a novel human placental tissue-derived biologic, PTP-001, which is in development as a candidate therapeutic for the treatment of osteoarthritis symptoms and pathophysiology. METHODS: Human placental tissues from healthy donors were prepared as a particulate formulation, PTP-001. PTP-001 extracts were assayed for the presence of disease-relevant biofactors which could have beneficial effects in treating osteoarthritis. PTP-001 eluates were tested in human chondrocyte cultures to determine effects on the production of a key collagen-degrading matrix metalloproteinase, MMP-13. PTP-001 eluates were also assessed for anti-inflammatory potential in human monocyte/macrophage cultures, as well as for growth-stimulating anabolic effects in human synoviocytes. The in vivo effects of PTP-001 on joint pain and histopathology were evaluated in a rat model of osteoarthritis induced surgically by destabilization of the medial meniscus. RESULTS: PTP-001 was found to contain an array of beneficial growth factors, cytokines and anti-inflammatory molecules. PTP-001 eluates dose-dependently inhibited the production of chondrocyte MMP-13, and the secretion of proinflammatory cytokines from monocyte/macrophage cultures. PTP-001 eluates also promoted proliferation of cultured synovial cells. In a rat osteoarthritis model, PTP-001 significantly reduced pain responses throughout 6 weeks post-dosing. The magnitude and duration of pain reduction following a single intraarticular treatment with PTP-001 was comparable to that observed for animals treated with a corticosteroid (active control). For rats dosed twice with PTP-001, significant reductions in cartilage histopathology scores were observed. CONCLUSIONS: PTP-001 represents a promising biologic treatment for osteoarthritis, with a multi-modal mechanism of action that may contribute to symptom management and disease modification.

Osteoarthritis induced by destabilization of the medial meniscus is reduced in germ-free mice.

OBJECTIVE: To determine the contribution of the gut microbiota to the development of injury-induced osteoarthritis (OA). DESIGN: OA was induced using the destabilized medial meniscus (DMM) model in 20 germ-free (GF) C57BL/6J male mice housed in a gnotobiotic facility and 23 strain-matched specific pathogen free (SPF) mice in 2 age groups -13.5 weeks avg age at DMM (17 SPF and 15 GF) and 43 weeks avg age at DMM (6 SPF and 5 GF). OA severity was measured using scores for articular cartilage structure (ACS), loss of safranin O (SafO) staining, osteophyte size, and synovial hyperplasia. Microbiome analysis by 16S rRNA amplicon sequencing was performed on stool samples and LPS and LPS binding protein (LBP) were measured in plasma. RESULTS: Compared to the SPF DMM mice, the maximum (MAX) ACS score per joint was 28% lower (p = 0.036) in GF DMM mice while the SafO sum score of all sections evaluated per joint was decreased by 31% (p = 0.009). The differences between SPF and GF mice in these scores were greater when only the younger mice were included in the analysis. The younger GF DMM mice also had significant reductions in osteophyte size (36%, P = 0.0119) and LBP (27%, P = 0.007) but not synovial scores or LPS. Differences in relative abundance of a number of Operational Taxonomic Units (OTUs) were noted between SPF mice with high vs low maximum ACS scores. CONCLUSIONS: These results suggest factors related to the gut microbiota promote the development of OA after joint injury.

Avian hosts of Ixodes pacificus (Acari: Ixodidae) and the detection of Borrelia burgdorferi in larvae feeding on the Oregon junco.

Larval and nymphal western blacklegged tick, Ixodes pacificus Cooley & Kohls (Acari: Ixodidae), were collected from birds, rodents, and lizards at Quail Ridge Reserve located in Napa County in northwestern California. Species from three vertebrate classes were sampled simultaneously from two transects during two consecutive spring seasons. Feeding larval and nymphal ticks were removed and preserved for counting, examination and testing for the presence of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner. Mean infestations with I. pacificus subadults on lizards were 10.0, on birds 2.9, and on rodents 1.3. I. pacificus larvae (204) collected from 10 avian species and (215) collected from two rodent species were tested for the presence of B. burgdorferi s.s. via real-time polymerase chain reaction. Three B. burgdorferi-infected larvae were taken from two Junco hyemalis and two infected larvae from one Neotoma fuscipes Baird. This is the detection of B. burgdorferi ss in an Ixodes pacificus larvae feeding on a Junco hyemalis L., [corrected] in western North America.

Influence of dietary fatty acid composition on the relationship between CETP activity and plasma lipoproteins in monkeys.

CETP activity, measured as transfer of cholesteryl ester from exogenous HDL to exogenous VLDL and LDL, reflecting CETP mass as determined by ELISA, was documented in three groups of St. Kitts vervet monkeys fed diets enriched in saturated (Sat), monounsaturated (Mono), or n-6 polyunsaturated (Poly) fatty acids. CETP activity was not different when comparing the three dietary fats. However, CETP activity was significantly higher when cholesterol was added to each of the diets. Significant positive associations between CETP activity and VLDL and LDL cholesterol concentrations were found whereas significant negative associations were seen between CETP activity and HDL cholesterol in each of the diet groups. The strength of these associations was highest in the Sat group. Cholesteryl ester (CE) fatty acid composition of lipoproteins varied widely among diet groups, with the more polyunsaturated CE of the Poly group being associated with a higher rate of CE transfer to endogenous acceptor apolipoprotein B-containing lipoproteins. Finally, only the Sat diet group showed significant positive correlations of CETP activity with LDL particle diameter (r = 0.76), cholesteryl ester percentage (r = 0.67), and a strong negative correlation (r = -0.86) with LDL receptor function, estimated as the difference between native and methylated LDL turnover rates. We speculate that strong associations between CETP and LDL metabolism may explain, at least in part, the increased atherogenicity of dietary saturated fat.

Partial characterization of retroviruses from boid snakes with inclusion body disease.

OBJECTIVE: To characterize retroviruses isolated from boid snakes with inclusion body disease (IBD). ANIMALS: 2 boa constrictors with IBD and 1 boa exposed to an affected snake. PROCEDURE: Snakes were euthanatized, and tissue specimens and blood samples were submitted for virus isolation. Tissue specimens were cultured with or without commercially available viper heart cells and examined by use of transmission electron microscopy (TEM) for evidence of viral replication. Reverse transcriptase activ ty was determined in sucrose gradient-purified virus. Western blotting was performed, using polyclonal antibodies against 1 of the isolated viruses. Specificity of the rabbit anti-virus antibody was evaluated, using an immunogold-labeling TEM technique. RESULTS: 3 viruses (RV-1, RV-2, and RV-3) were isolated. The isolates were morphologically comparable to members of the Retroviridae family. Reverse transcriptase activity was high in sucrose gradient fractions that were rich in virus. Polyclonal antibody against RV-1 reacted with proteins of similar relative mobility in RV-1 and RV-2. By use of immunogold labeling, this antibody also recognized virions of both RV-1 and RV-2. CONCLUSIONS AND CLINICAL RELEVANCE: A retrovirus was isolated from boid snakes with IBD or exposed to IBD. Western blot analysis of viral proteins indicated that viruses isolated from the different snakes were similar. Whether this virus represents the causative agent of IBD is yet to be determined. The isolation of retroviruses from boid snakes with IBD is an important step n the process of identifying the causative agent of this disease.

Co-isolation of heat stress and cytoskeletal proteins with plasma membrane proteins.

Previous reports have suggested the possible existence of a plasma-cell-membrane function associated with some heat stress proteins (HSPs). To investigate the effect of hyperthermia on plasma membrane proteins, rat mammary tumour clone C (MTC) cells were heated at 42 degrees C for 1 h. Their surface proteins were (1) labelled with [3H]leucine, (2) biotinylated, (3) affinity isolated with streptavidin-agarose beads under denaturing or non-denaturing conditions, and (4) analysed by one- and two-dimensional polyacrylamide-gel electrophoresis and protein blotting under denaturing conditions. Affinity isolation of biotinylated proteins enriched for a protein subfraction believed to be membrane-associated. Several proteins analogous to HSP or their heat-stress cognates (HSC) were present with these biotinylated protein subfractions in control or heated cells. The major and most consistent feature of affinity isolates from heated cells was the presence of a small fraction of the induced 68-kD HSP. The 112-, 90-, 70- and 22-kD HSC/HSP were also present in small amounts in affinity isolates of control cells, and the fraction increased in heated cells. Several structural proteins, including actin and the tubulins were present in the same affinity isolates. Protein blotting experiments indicated that none were exposed on the exterior of the plasma-cell membrane or biotinylated and thus none were exposed on the exterior of the plasma-cell membrane or biotinylated intracellularly through membrane damage. These results suggest that small fractions of several HSC are located at or near the cytoplasmic face of the plasma membrane along with cytoskeletal proteins, and that additional submembranous localization of HSP occurs after heat stress and may be part of the processes associated with membrane damage or cellular responses to heat. Further studies will be directed at establishing the relationships between these proteins and the role, if any, of the changes associated with heat stress.