Bone marrow mononuclear cells for joint therapy: The role of macrophages in inflammation resolution and tissue repair

Osteoarthritis(OA)is the most prevalent joint disease causing major disability and medical expenditures.Synovitis is a central feature of OA and is primarily driven by macrophages.Synovial macrophages not only drive inflammation but also its resolution,through a coordinated,simultaneous expression of pro-and anti-inflammatory mechanisms that are essential to counteract damage and recover homeostasis.Current OA therapies are largely based on anti-inflammatory principles and therefore block pro-inflammatory mechanisms such as prostaglandin E2 and Nuclear factor-kappa B signaling pathways.However,such mechanisms are also innately required for mounting a pro-resolving response,and their blockage often results in chronic low-grade inflammation.Following minor injury,macrophages shield the damaged area and drive tissue repair.If the damage is more extensive,macrophages incite inflammation recruiting more macrophages from the bone marrow to maximize tissue repair and ultimately resolve inflammation.However,sustained damage and inflammation often overwhelms pro-resolving mechanisms of synovial macrophages leading to the chronic inflammation and related tissue degeneration observed in OA.Recently,experimental and clinical studies have shown that joint injection with autologous bone marrow mononuclear cells replenishes inflamed joints with macrophage and hematopoietic progenitors,enhancing mechanisms of inflammation resolution,providing remarkable and long-lasting effects.Besides creating an ideal environment for resolution with high concentrations of interleukin-10 and anabolic growth factors,macrophage progenitors also have a direct role in tissue repair.Macrophages constitute a large part of the early granulation tissue,and further transdifferentiate from myeloid into a mesenchymal phenotype.These cells,characterized as fibrocytes,are essential for repairing osteochondral defects.Ongoing“omics”studies focused on identifying key drivers of macrophagemediated resolution of joint inflammation and those required for efficient osteochondral repair,have the potential to uncover ways for developing engineered macrophages or off-the-shelf pro-resolving therapies that can benefit patients suffering from many types of arthropaties,not only OA.

Diclazuril Protects against Maternal Gastrointestinal Syndrome and Congenital Toxoplasmosis

Background: Toxoplasmosis is a common cause of foodborne, gastrointestinal and congenital syndrome with particularly severe or unknown health consequences. There is no safe and effective preventive or therapeutic modality against congenital toxoplasmosis or to eliminate the persistent chronic infection. Hypothesis: Diclazuril to be safe in pregnancy and effective against gastrointestinal toxoplasmosis. Methods: CD1 programmed pregnant mice were divided into groups and administered a diet containing diclazuril, or sham control. Treatments were initiated on Day 5 of pregnancy and continued until Day 16 when dams were euthanatized. On Day 8 of pregnancy dams were infected intraperitoneally with escalating doses of tachyzoites (0, 100, 300, 600) from Type II strain. Dams were monitored daily for distress, pain, and abortion and samples collected at the end of the experiments. Results: Infected dams developed moderate to severe Toxoplasma related complications in tachyzoites dose dependent manner. Animals became anemic and showed hydrothorax, and ascities. Diclazuril effectively protected dams from ascities and anemia (p < 0.05). Infected dams showed splenomegaly, with massive infiltration of epithelioid cells compared with the protective effect of diclazuril in treated animals. Infected dams exhibited severe hepatitis (score 0 to 4 scale = 3.5 ± 0.01) with influx of inflammatory and plasma cells, dysplastic hepatocytes, multinucleated giant cell transformation and hepatic cells necrosis. Diclazuril treatment significantly protected dams from hepatitis, also in tachyzoites dose (100, 300, 600) dependent manner (respectively infected-treated versus infected controls, p < 0.001, p < 0.01 and p with infiltration of lymphocytes, and macrophages and microabscess formations in the cryptic structures, with significant improvement in diclazuril treated animals. Additionally, the number of fetuses, fetal length and fetal weight were preserved in diclazuril treated dams. Conclusions: This is the first report describing of diclazuril safety in pregnancy as well as efficacy against mild to moderate hepato-gastrointestinal syndrome in dams and fetal toxoplasmosis (Special issue, “Treatment of Liver Diseases”).

Clinical and Radiographic Evaluation of Cattle Tail before and after the <i>Vaquejada</i>Race

The vaquejada race is a sport with animals which has the greatest socio-economic impact in Brazil and is responsible for generating an intense cultural and economic movement. However, this activity has been the subject of questions about well-being involving the environmental and mental comfort conditions offered to equine and cattle interactions. It is necessary to perform technical evaluations of the cattle involved on the basis of scientific study to better understand the processes of sport and, with this, promote their evolution on good welfare practices in the 21st century. Therefore, to test the hypothesis that the use of the official rules of the vaquejada where cattle use a protective harness on the tail protects the cattle’s tail from injuries in this region that compromise health. Before, during and after races, a study was developed that aimed to perform a general and radiological clinical evaluation in the proximal coccygeal region in cattle that ran in a modern cattle stall. Forty male bovines were evaluated (average: 2 year-old and 420 kg). Oxen were subjected to clinical and radiological examination before and after racing with official rules. Results of the clinical and radiological evaluations performed by veterinarians, did not observe clinical occurrences before and after the races in the 40 cattle evaluated. The increase in heart and respiratory rates was a reflection of temporary management stressors, as the animals returned to the feeding/rumination/interaction pattern after running in the rest pen. The radiographic findings in the tail, represented by the reduction of the intervertebral space and areas of subcutaneous radiolucency, suggest technical artifacts derived from the manipulation of the tail for proper positioning to perform the tests in the condition of retention in the corral. The partial fracture found by an evaluator showed signs of calcification and was accompanied by swelling in the soft tissue but without pain in the pre-run examination. It is concluded that the modern vaquejada race, which meets the official rules, is not related to the increase in lesions in the locomotor apparatus or other evaluated tissues, including the tail.

Effort and Recovery in Nellore Oxen during Vaquejada Assessed with Ocular and Tail Infrared Thermography Superficial Temperature

Infrared thermography (IRT) has emerged for evaluation of animal welfare. To test the hypothesis that cattle subjected to vaquejada increased temperature in the eye (CLO) and tail due to physical effort, a research was developed that aimed to measure maximum surface temperature in the CLO and base of tail using IRT. Eighty Nellore cattle were used, which were subjected to physical effort in three periods (morning, afternoon and night). IRT was performed at CLO and base of tail, in rest pen/corral (control) and during the vaquejada (pre- and post-run). Tails’ analysis was divided in 3 points (E1, E2 and E3). ANOVA and Tukey’s test (p < 0.001) were used for analysis. Temperatures were higher in morning and afternoon and different from those at night on control (p < 0.001). During vaquejada with one run, CLO was higher in pre-run, followed by a slight reduction in post-run (p < 0.001). Analyzing temperatures variations at tail, higher temperatures were observed in the morning, followed by the evening and night (p < 0.001), but without differences within the pre- and post-run periods. Temperatures at tail’s points were higher in morning period in cattle with two runs (p < 0.001). Finally, it was concluded that there were no increases in CLO or at tail’s points after vaquejada races. The IRT method was efficient in determining surface temperatures in CLO and tail points in cattle under the same conditions and may be a good noninvasive method for clinical and welfare assessments.

Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2

Leptospirosis is an infectious bacterial disease caused by Leptospira species. In this study, we cloned and se- quenced the gene encoding the immunodominant protein GroEL from L. interrogans serovar Autumnalis strain N2, which was isolated from the urine of a patient during an outbreak of leptospirosis in Chennai, India. This groEL gene encodes a protein of 60 kDa with a high degree of homology （99% similarity） to those of other leptospiral serovars. Recombinant GroEL was overexpressed in Escherichia coli. lmmunoblot analysis indi- cated that the sera from confirmed leptospirosis patients showed strong reactivity with the recombinant GroEL while no reactivity was observed with the sera from seronegative control patient. In addition, the 3D structure of GroEL was constructed using chaperonin complex cpn60 from Thermus thermophilus as template and vali- dated. The results indicated a Z-score of -8.35, which is in good agreement with the expected value for a pro- tein. The superposition of the Ca traces of cpn60 structure and predicted structure of leptospiral GroEL indicates good agreement of secondary structure elements with an RMSD value of 1.5A. Further study is necessary to evaluate GroEL for serological diagnosis of leptospirosis and for its potential as a vaccine component.