Comprehensive Physicochemical and Biological Characterization of the Proposed Biosimilar Darbepoetin Alfa, LBDE, and Its Originator Darbepoetin Alfa, NESP®.

BACKGROUND: For regulatory approval, the comparability of a biosimilar product to an originator product should be ensured through thorough physicochemical and biological characterization. OBJECTIVE: To evaluate the biosimilarity between LBDE, the proposed biosimilar darbepoetin alfa, and NESP®, its originator, we performed a comprehensive physicochemical and biological characterization study. METHODS: Primary and higher-order protein structures were analyzed using Lys-C peptide mapping with liquid chromatography-mass spectrometry (LC-MS), disulfide bond identification, circular dichroism, and fluorescence spectroscopy. Glycosylation and isoform distribution were analyzed using MS, LC, and capillary zone electrophoresis. Size variants were evaluated with size-exclusion chromatography-high-performance liquid chromatography (SEC-HPLC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Biological characterization included binding affinity for human erythropoietin receptor, in vitro cell proliferation, and in vivo potency. Pharmacokinetics (PK) were evaluated using rats through two injection routes. RESULTS: Non-reducing and reducing Lys-C peptide mapping showed a highly similar peak profile, confirming that LBDE and NESP® have the same primary structure and disulfide bonds. Glycosylation and isoform analyses showed that the attached N-glycan and O-glycan structures were the same and their relative contents were similar. Spectroscopic analysis of LBDE showed indistinguishable spectra with NESP®. For both LBDE and NESP®, a very small amount of size variants was found in SEC-HPLC, and no minor bands were detected in SDS-PAGE. Furthermore, LBDE did not show any difference with NESP® in the in vitro and in vivo functional analyses. PK parameters of LBDE were in good agreement with those of NESP®. CONCLUSION: LBDE shows high similarity to NESP® with regard to structure and function.

Zanthoxylum schinifolium leaf ethanol extract inhibits adipocyte differentiation through inactivation of the extracellular signal regulated kinase and phosphoinositide 3-kinase/Akt signaling pathways in 3T3-L1 pre-adipocytes.

Zanthoxylum schinifolium is widely used as a food flavoring in east Asia. Although this plant has also been used in traditional oriental medicine for the treatment of the common cold, toothache, stomach ache, diarrhea and jaundice, its anti-obesity activity remains to be elucidated. The present study investigated the effects of ethanol extract from the leaves of Z. schinifolium (EEZS) on adipocyte differentiation, and its underlying mechanism, in 3T3-L1 pre-adipocytes. The results demonstrated that EEZS effectively suppressed intracellular lipid accumulation at non-toxic concentrations, and was associated with the downregulation of several adipocyte-specific transcription factors, including peroxisome proliferation-activity receptor γ (PPARγ), CCAAT/enhancer binding protein (C/EBP)α and C/EBPß, in a concentration-dependent manner. Furthermore, it was observed that EEZS markedly inactivated the extracellular signal-regulated protein kinase (ERK) and phosphatidylinositide 3-kinase (PI3K)/Akt pathways, which act upstream of PPARγ and C/EBPs in adipogenesis. These results suggested that EEZS inhibited lipid accumulation by downregulating the major transcription factors involved in the pathway of adipogenesis, including PPARγ, C/EBPα and C/EBPß, via regulation of the ERK and PI3K/Akt signaling pathways in 3T3-L1 adipocyte differentiation. This indicated the potential use of EEZS as an anti-obesity agent.

The effects of kinesio taping on architecture, strength and pain of muscles in delayed onset muscle soreness of biceps brachii.

[Purpose] This study aimed to confirm the effects of kinesio taping (KT) on muscle function and pain due to delayed onset muscle soreness (DOMS) of the biceps brachii. [Subjects and Methods] Thirty-seven subjects with induced DOMS were randomized into either Group I (control, n=19) or Group II (KT, n=18). Outcome measures were recorded before the intervention (application of KT) and at 24, 48, and 72 hours after the intervention. DOMS was induced, and muscle thickness was measured using ultrasonic radiography. Maximal voluntary isometric contraction (%MVIC) was measured via electromyography (EMG). Subjective pain was measured using a visual analogue scale (VAS). [Results] Group I exhibited a positive correlation between muscle thickness and elapsed time from intervention (24, 48, and 72 hours post induction of DOMS); they also showed a significant decrease in MVIC(%). Group II showed significant increases in muscle thickness up to the 48-hour interval post induction of DOMS, along with a significant decrease in MVIC (%). However, in contrast to Group I, Group II did not show a significant difference in muscle thickness or MVIC (%) at the 72-hour interval in comparison with the values prior to DOMS induction. [Conclusion] In adults with DOMS, activation of muscles by applying KT was found to be an effective and faster method of recovering muscle strength than rest alone.

The effects of sensorimotor training on anticipatory postural adjustment of the trunk in chronic low back pain patients.

[Purpose] This study aimed to examine the effects of sensorimotor training on the anticipatory postural adjustment (APA) of chronic low back pain (CLBP) patients. [Subjects and Methods] Fourteen CLBP patients were randomly assigned to Group II (ordinary physical therapy, n=7) and Group III (sensorimotor training, n=7). In addition, a normal group (Group I) consisting of seven subjects was chosen as the control group. The two CLBP groups received their own treatment five times per week, for four weeks, for 40 minutes each time. Changes in pain and functional performance evaluation were examined by the visual analogue scale (VAS) and the Oswestry Disability Index (ODI). In order to look at the change in APA, muscle onset time was examined using electromyography (EMG). [Results] Group III showed significant changes in both VAS and ODI. According to comparison of the results for muscle onset time, there were significant decreases in Group III's transversus abdominis muscle (TrA) and external oblique muscle (EO) in the standing and sitting positions. There were significant differences between Group II and III in terms of the TrA in the sitting position. [Conclusion] Sensorimotor training makes patients capable of learning how to adjust muscles, thereby alleviating pain and improving muscle performance.

A case of portal vein thrombosis by protein C and s deficiency completely recanalized by anticoagulation therapy.

Portal vein thrombosis (PVT) is a rare form of venous thrombosis that affects the hepatic portal vein flow, which can lead to portal hypertension. Treatment of PVT includes anticoagulants, thrombolysis, insertion of shunts, bypass surgery, and liver transplantation. Single anticoagulation therapy is not regarded as a curative treatment but can be associated with a reduction in new thrombotic episodes. We experienced a case of acute total occlusion of PVT provoked by protein C and S deficiency syndrome. PVT was completely recanalized with oral anticoagulant therapy following low molecular weight heparin therapy.