The purpose of this study was to evaluate the effects of sclerostin inhibition by treatment with a sclerostin antibody (Scl-AbII) on bone formation, bone mass, and bone strength in an aged, gonad-intact male rat model. Sixteen-month-old male Sprague-Dawley rats were injected subcutaneously with vehicle or Scl-AbII at 5 or 25 mg/kg twice per week for 5 weeks (9-10/group). In vivo dual-energy X-ray absorptiometry (DXA) analysis showed that there was a marked increase in areal bone mineral density of the lumbar vertebrae (L 1 to L 5 ) and long bones (femur and tibia) in both the 5 and 25 mg/kg Scl-AbII-treated groups compared with baseline or vehicle controls at 3 and 5 weeks after treatment. Ex vivo micro-computed tomographic (mCT) analysis demonstrated improved trabecular and cortical architecture at the fifth lumbar vertebral body (L 5 ), femoral diaphysis (FD), and femoral neck (FN) in both Scl-AbII dose groups compared with vehicle controls. The increased cortical and trabecular bone mass was associated with a significantly higher maximal load of L 5 , FD, and FN in the high-dose group. Bone-formation parameters (ie, mineralizing surface, mineral apposition rate, and boneformation rate) at the proximal tibial metaphysis and tibial shaft were markedly greater on trabecular, periosteal, and endocortical surfaces in both Scl-AbII dose groups compared with controls. These results indicate that sclerostin inhibition by treatment with a sclerostin antibody increased bone formation, bone mass, and bone strength in aged male rats and, furthermore, suggest that pharmacologic inhibition of sclerostin may represent a promising anabolic therapy for low bone mass in aged men. ß

Background-Preeclampsia is a serious disorder of pregnancy characterized by hypertension, proteinuria, edema, and coagulation and vascular abnormalities. At the cellular level, abnormalities include increased calcium concentration in platelets, lymphocytes, and erythrocytes. Recent studies have shown that antibodies directed against angiotensin II type I (AT 1 ) receptors are also highly associated with preeclampsia. Methods and Results-We tested the hypothesis that AT 1 receptor-agonistic antibodies (AT 1 -AAs) could activate AT 1 receptors, leading to an increased intracellular concentration of free calcium and to downstream activation of Ca 2ϩ signaling pathways. Sera of 30 pregnant patients, 16 diagnosed with severe preeclampsia and 14 normotensive, were examined for the presence of IgG capable of stimulating intracellular Ca 2ϩ mobilization. IgG from all preeclamptic patients activated AT 1 receptors and increased intracellular free calcium. In contrast, none of the normotensive individuals had IgG capable of activating AT 1 receptors. The specific mobilization of intracellular Ca 2ϩ by AT 1 -AAs was blocked by losartan, an AT 1 receptor antagonist, and by a 7-amino-acid peptide that corresponds to a portion of the second extracellular loop of the AT 1 receptor. In addition, we have shown that AT 1 -AA-stimulated mobilization of intracellular Ca 2ϩ results in the activation of the transcription factor, nuclear factor of activated T cells. Conclusions-These results suggest that maternal antibodies capable of activating AT 1 receptors are likely to account for increased intracellular free Ca 2ϩ concentrations and changes in gene expression associated with preeclampsia. Key Words: autoimmunity Ⅲ antibodies Ⅲ signal transduction Ⅲ hypertension Ⅲ pregnancy P reeclampsia is a pregnancy-induced hypertensive disorder that affects Ϸ3% to 5% of first pregnancies and is a leading cause of maternal and fetal mortality and morbidity. This disorder is characterized by the development of a maternal syndrome that includes hypertension, coagulation abnormalities, edema, proteinuria, and vascular abnormalities. These symptoms normally develop after 20 weeks of gestation and disappear within 7 to 10 days after delivery. However, subtle signs, such as increased protein excretion, may persist for several months. The main target organs affected in preeclampsia are the brain, kidneys, liver, lungs, placenta, and heart. Advanced stages of the disease include cerebral hemorrhage, renal failure, and the HELLP syndrome (hemolysis, elevated liver enzymes, and low platelets). Two well-known pathogenic features of preeclampsia are placental and vascular abnormalities. Shallow trophoblast invasion and improper remodeling of spiral arteries are among the bestrecognized and most frequently associated features of this disease. Altered levels of circulating vasoactive substances, such as endothelin and prostaglandins, have been reported in preeclampsia. 1 Preeclampsia is also associated with abnormalities in Ca 2ϩ metabolism and increased ...

The 2015 9th Workshop on Recent Issues in Bioanalysis (9th WRIB) took place in Miami, Florida with participation of 600 professionals from pharmaceutical and biopharmaceutical companies, biotechnology companies, contract research organizations and regulatory agencies worldwide. WRIB was once again a 5 day, week-long event - A Full Immersion Bioanalytical Week - specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest in bioanalysis. The topics covered included both small and large molecules, and involved LCMS, hybrid LBA/LCMS and LBA approaches, including the focus on biomarkers and immunogenicity. This 2015 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2015 edition of this comprehensive White Paper has been divided into three parts. Part 3 discusses the recommendations for large molecule bioanalysis using LBA, biomarkers and immunogenicity. Part 1 (small molecule bioanalysis using LCMS) and Part 2 (hybrid LBA/LCMS and regulatory inputs from major global health authorities) have been published in volume 7, issues 22 and 23 of Bioanalysis, respectively.

The 2016 10th Workshop on Recent Issues in Bioanalysis (10th WRIB) took place in Orlando, Florida with participation of close to 700 professionals from pharmaceutical/biopharmaceutical companies, biotechnology companies, contract research organizations, and regulatory agencies worldwide. WRIB was once again a weeklong event - A Full Immersion Week of Bioanalysis for PK, Biomarkers and Immunogenicity. As usual, it is specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest including both small and large molecules involving LCMS, hybrid LBA/LCMS, and LBA approaches, with the focus on PK, biomarkers and immunogenicity. This 2016 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. This White Paper is published in 3 parts due to length. This part (Part 3) discusses the recommendations for large molecule bioanalysis using LBA, biomarkers and immunogenicity. Parts 1 (small molecule bioanalysis using LCMS) and Part 2 (Hybrid LBA/LCMS and regulatory inputs from major global health authorities) have been published in the Bioanalysis journal, issues 22 and 23, respectively.

The 2013 7th Workshop on Recent Issues in Bioanalysis was held in Long Beach, California, USA, where close to 500 professionals from pharmaceutical and biopharmaceutical companies, CROs and regulatory agencies convened to discuss current topics of interest in bioanalysis. These 'hot' topics, which covered both small and large molecules, were the starting point for fruitful exchanges of knowledge, and sharing of ideas among speakers, panelists and attendees. The discussions led to specific recommendations pertinent to bioanalytical science. Such as the previous editions, this 2013 White Paper addresses important bioanalytical issues and provides practical answers to the topics presented, discussed and agreed upon by the global bioanalytical community attending the 7th Workshop on Recent Issues in Bioanalysis.