Loganin Prevents Hepatic Steatosis by Blocking NLRP3 Inflammasome Activation

Activation of the NLRP3 inflammasome is a necessary process to induce fibrosis in nonalcoholic fatty liver disease (NAFLD). Nonalcoholic steatohepatitis (NASH) is a kind of NAFLD that encompasses the spectrum of liver disease. It is characterized by inflammation and ballooning of hepatocytes during steatosis. We tested whether inhibiting the NLRP3 inflammasome could prevent the development and pathology of NASH. We identified loganin as an inhibitor of the NLRP3 inflammasome and investigated whether in vivo administration of loganin prevented NASH symptoms using a methionine-choline deficient (MCD) diet model in mice. We found that loganin inhibited the NLRP3 inflammasome activation triggered by ATP or nigericin, as shown by suppression of the production of interleukin (IL)-1β and caspase-1 (p10) in mouse primary macrophages. The speck formation of apoptosisassociated speck-like protein containing a caspase recruitment domain (ASC) was blocked by loganin, showing that the assembly of the NLRP3 inflammasome complex was impaired by loganin. Administration of loganin reduced the clinical signs of NASH in mice fed the MCD diet, including hepatic inflammation, fat accumulation, and fibrosis. In addition, loganin reduced the expression of NLRP3 inflammasome components in the liver. Our findings indicate that loganin alleviates the inflammatory symptoms associated with NASH, presumably by inhibiting NLRP3 inflammasome activation. In summary, these findings imply that loganin may be a novel nutritional and therapeutic treatment for NASH-related inflammation.

Prediction of the Duration to Next Admission for an Acute Affective Episode in Patients with Bipolar I Disorder

Objective@#Predicting disease relapse and early intervention could reduce symptom severity. We attempted to identify potential indicators that predict the duration to next admission for an acute affective episode in patients with bipolar I disorder. @*Methods@#We mathematically defined the duration to next psychiatric admission and performed single-variate regressions using historical data of 101 patients with bipolar I disorder to screen for potential variables for further multivariate regressions. @*Results@#Age of onset, total psychiatric admissions, length of lithium use, and carbamazepine use during the psychiatric hospitalization contributed to the next psychiatric admission duration positively. The all-in-one found that hyperlipidemia during the psychiatric hospitalization demonstrated a negative contribution to the duration to next psychiatric admission; the last duration to psychiatric admission, lithium and carbamazepine uses during the psychiatric hospitalization, and heart rate on the discharge day positively contributed to the duration to next admission. @*Conclusion@#We identified essential variables that may predict the duration of bipolar I patients’ next psychiatric admission. The correlation of a faster heartbeat and a normal lipid profile in delaying the next onset highlights the importance of managing these parameters when treating bipolar I disorder.

Brain endothelial cell-derived extracellular vesicles with a mitochondria-targeting photosensitizer effectively treat glioblastoma by hijacking the blood‒brain barrier

Glioblastoma (GBM) is the most aggressive malignant brain tumor and has a high mortality rate. Photodynamic therapy (PDT) has emerged as a promising approach for the treatment of malignant brain tumors. However, the use of PDT for the treatment of GBM has been limited by its low blood‒brain barrier (BBB) permeability and lack of cancer-targeting ability. Herein, brain endothelial cell-derived extracellular vesicles (bEVs) were used as a biocompatible nanoplatform to transport photosensitizers into brain tumors across the BBB. To enhance PDT efficacy, the photosensitizer chlorin e6 (Ce6) was linked to mitochondria-targeting triphenylphosphonium (TPP) and entrapped into bEVs. TPP-conjugated Ce6 (TPP-Ce6) selectively accumulated in the mitochondria, which rendered brain tumor cells more susceptible to reactive oxygen species-induced apoptosis under light irradiation. Moreover, the encapsulation of TPP-Ce6 into bEVs markedly improved the aqueous stability and cellular internalization of TPP-Ce6, leading to significantly enhanced PDT efficacy in U87MG GBM cells. An in vivo biodistribution study using orthotopic GBM-xenografted mice showed that bEVs containing TPP-Ce6 [bEV(TPP-Ce6)] substantially accumulated in brain tumors after BBB penetration via transferrin receptor-mediated transcytosis. As such, bEV(TPP-Ce6)-mediated PDT considerably inhibited the growth of GBM without causing adverse systemic toxicity, suggesting that mitochondria are an effective target for photodynamic GBM therapy.

Biomechanical study on different high-strength sutures and suture site for repairing posterior root tear of the medial meniscus / 中国骨伤

OBJECTIVE@#To compare biomechanical characteristic of different high-strength sutures and suture sites for repairing posterior root tear of the medial meniscus with modified Mason-Allen technique.@*METHODS@#Forty-eight specimen of medial meniscus of knee joint from fresh porcine (female, aged from 5 to 9 months with an average of 7 months) were chosen and established experimental model. The samples were divided into red zone fixation group and red-white zone fixation group according to suture sites, 24 in each group; and then were randomly divided into 3 subgroups which 8 in each group, and fixed with Ethibond suture, Ultrabraid suture and FiberWire suture, respectively. Biomechanical tests were performedon universal electromagnetic and mechanical testing machine. Each specimen was underwent 1 000 cyclic tests on the first time, then pull out test until failure. The maximum failure load, yield load, stiffness and displacement were analyzed.@*RESULTS@#All specimen were successfully completed biomechanical tests. The failure mode of Ethibond group was caused by suture fracture; 6 cases of Ultrabraid suture group was caused by suture fracture which belong to red zone fixation group, 10 cases were caused by suture pull out, which 2 cases belong to red zone fixation group, 8 cases belong to red-white zone fixation group;8 cases of FiberWire group was caused by suture pull-out. Biomechanical test showed that:(1)In terms of suture strength, comparison of the maximum failure load, yield load and stiffness showed that Ethibond suture group

Radiological Assessment of the Effects of Anterior Cervical Discectomy and Fusion on Distraction of the Posterior Ligamentum Flavum in Patients with Degenerative Cervical Spines

Background@#This study aimed to assess the effects of anterior cervical discectomy and fusion (ACDF) on distraction of the posterior ligamentum flavum (LF) by increasing the intervertebral disc height and positioning a graft in patients with degenerative cervical spine disease. @*Methods@#Sixty-eight patients with degenerative cervical diseases who underwent single-level ACDF were included in the analysis. The intervertebral disc height, Cobb angle, and transverse thickness of the LF were measured, and magnetic resonance imaging was performed both preoperatively and 6 weeks postoperatively on each patient. Correlation analyses were performed to evaluate the relationships between age, sex, change in intervertebral disc height, Cobb angle, and position of the intervertebral implant according to the postoperative change in LF thickness. The position of the intervertebral implant was categorized as anterior, middle, or posterior. We also evaluated radiological effects according to the implant position. @*Results@#The mean intervertebral disc height increased from 5.88 mm preoperatively to 7.49 mm postoperatively. The Cobb angle was 0.88° preoperatively and 1.43° postoperatively. Age (p = 0.551), sex (p = 0.348), position of cage (p = 0.312), pre- and postoperative intervertebral disc height (p = 0.850, p = 0.900), Cobb angle (p = 0.977, p = 0.460), and LF thickness (p = 0.060, p = 1.00) were not related to changes in postoperative LF thickness. Postoperative increase in disc height was related to Cobb angle (r = 0.351, p = 0.038). No other factors were significantly related. The position of the cage was not related with the change of Cobb angle (p = 0.91), LF thickness (p = 0.31), or disc height (p = 0.54). @*Conclusions@#Change in the intervertebral disc height and the position of the intervertebral implant after ACDF did not affect the thickness of the LF after surgery in patients with degenerative cervical spine disease.

F-box Protein βTrCP1 Is a Substrate of Extracellular Signal-regulated Kinase 2

F-box proteins, consisting of 69 members which are organized into the three subclasses FBXW, FBXL, and FBXO, are the substrate specific recognition subunits of the SKP1-Cullin 1-F-box protein E3 ligase complex. Although βTrCP 1 and 2, members of the FBXW subfamily, are known to regulate some protein stability, molecular mechanisms by which these proteins can recognize proper substrates are unknown. In this study, it was found that βTrCP1 showed strong interaction with members of mitogen-activated protein kinases. Although extracellular signal-regulated kinase (ERK) 3, p38β, and p38δ showed weak interactions, ERK2 specifically interacted with βTrCP1 as assessed by immunoprecipitation. In interaction domain determination experiments, we found that ERK2 interacted with two independent ERK docking sites located in the F-box domain and linker domain, but not the WD40 domain, of βTrCP1. Notably, mutations of βTrCP1 at the ERK docking sites abolished the interaction with ERK2. βTrCP1 underwent phosphorylation by EGF stimulation, while the presence of the mitogen-activated protein kinase kinases inhibitor U0126, genetic silencing by sh-ERK2, and mutation of the ERK docking site of βTrCP1 inhibited phosphorylation. This inhibition of βTrCP1 phosphorylation resulted in a shortened half-life and low protein levels. These results suggest that ERK2-mediated βTrCP1 phosphorylation may induce the destabilization of βTrCP1.

F-box Protein βTrCP1 Is a Substrate of Extracellular Signal-regulated Kinase 2

F-box proteins, consisting of 69 members which are organized into the three subclasses FBXW, FBXL, and FBXO, are the substrate specific recognition subunits of the SKP1-Cullin 1-F-box protein E3 ligase complex. Although βTrCP 1 and 2, members of the FBXW subfamily, are known to regulate some protein stability, molecular mechanisms by which these proteins can recognize proper substrates are unknown. In this study, it was found that βTrCP1 showed strong interaction with members of mitogen-activated protein kinases. Although extracellular signal-regulated kinase (ERK) 3, p38β, and p38δ showed weak interactions, ERK2 specifically interacted with βTrCP1 as assessed by immunoprecipitation. In interaction domain determination experiments, we found that ERK2 interacted with two independent ERK docking sites located in the F-box domain and linker domain, but not the WD40 domain, of βTrCP1. Notably, mutations of βTrCP1 at the ERK docking sites abolished the interaction with ERK2. βTrCP1 underwent phosphorylation by EGF stimulation, while the presence of the mitogen-activated protein kinase kinases inhibitor U0126, genetic silencing by sh-ERK2, and mutation of the ERK docking site of βTrCP1 inhibited phosphorylation. This inhibition of βTrCP1 phosphorylation resulted in a shortened half-life and low protein levels. These results suggest that ERK2-mediated βTrCP1 phosphorylation may induce the destabilization of βTrCP1.

Fractures of implant fixtures: a retrospective clinical study / 대한악안면성형재건외과학회지

Background@#The aim of this study was to evaluate the factors that may affect implant fixture fractures. @*Methods@#Patients who experienced implant fixture removal at Seoul National University Bundang Hospital from 2007 to 2015 due to implant fixture fracture were included. Implant/crown ratio, time of implant fracture, clinical symptoms before implant fracture, treatment of fractured implants, and the success and survival rate of the replaced implants were evaluated retrospectively. @*Results@#Thirteen implants were fractured in 12 patients. Patient mean age at the time of fracture was 59.3 years. Of the 13 implants, 7 implants were placed at our hospital, and 6 were placed at a local clinic. The mean crown/ implant ratio was 0.83:1. The clinical symptoms before fracture were screw loosening in five implants, marginal bone loss in five implants, and the presence of peri-implant diseases in five implants. All the fractured implants were removed, and 12 out of the 13 sites were re-implanted. Parafunctions were observed in two patients: one with bruxism and one with attrition due to a strong chewing habit. @*Conclusions@#Several clinical symptoms before the fracture of an implant can predict implant fixture failure. Therefore, if these clinical symptoms are observed, appropriate treatments can be taken before more serious complications result.

Repurposing Auranofin, an Anti-Rheumatic Gold Compound, to Treat Acne Vulgaris by Targeting the NLRP3 Inflammasome

Activation of the NLRP3 inflammasome is critical for host defense as well as the progression of inflammatory diseases through the production of the proinflammatory cytokine IL-1β, which is cleaved by active caspase-1. It has been reported that overactivation of the NLRP3 inflammasome contributes to the development and pathology of acne vulgaris. Therefore, inhibiting activation of the NLRP3 inflammasome may provide a new therapeutic strategy for acne vulgaris. In this study, we investigated whether auranofin, an anti-rheumatoid arthritis agent, inhibited NLRP3 inflammasome activation, thereby effectively treating acne vulgaris.Auranofin suppressed NLRP3 inflammasome activation induced by Propionibacterium acnes, reducing the production of IL-1β in primary mouse macrophages and human sebocytes. In a P. acnes-induced acne mouse model, injection of P. acnes into the ears of mice induced acne symptoms such as redness, swelling, and neutrophil infiltration. Topical application of auranofin (0.5 or 1%) to mouse ears significantly reduced the inflammatory symptoms of acne vulgaris induced by P. acnes injection. Topical application of auranofin led to the downregulation of the NLRP3 inflammasome activated by P. acnes in mouse ear skin. These results show that auranofin inhibits the NLRP3 inflammasome, the activation of which is associated with acne symptoms. The results further suggest that topical application of auranofin could be a new therapeutic strategy for treating acne vulgaris by targeting the NLRP3 inflammasome.

Implant Placement Using Palatal Bone in Patients with Severe Maxillary Alveolar Bone Defect: Case Series Study

Purpose@#The purpose of this case study series was to introduce successful implant cases that used the palatal bone in patients with severe maxillary alveolar bone atrophy or defects.Case Presentation: In this case series study, a total of four patients underwent implant placement in the palatal bone of the maxilla. A total of 6 implants were installed using the palatine bone. The patients’ ages ranged from 40 to 73 years with an average age of 63.5 years. The patients had maxillary sinus-related diseases, such as maxillary sinusitis, oroantral fistula, and dentigerous cysts, prior to implantation. To achieve initial stability, the implants were placed on the palatal side, and buccally tilted. The average postoperative recovery period was 8 months. No postoperative complications occurred in any of the cases, and the approach was used without reported issues. @*Conclusion@#Implant treatment by securing the initial fixation in the palatal bone is a good alternative when an implant must be installed in a patient who requires extensive and invasive bone graft.

Implant Placement Using Palatal Bone in Patients with Severe Maxillary Alveolar Bone Defect: Case Series Study

Purpose@#The purpose of this case study series was to introduce successful implant cases that used the palatal bone in patients with severe maxillary alveolar bone atrophy or defects.Case Presentation: In this case series study, a total of four patients underwent implant placement in the palatal bone of the maxilla. A total of 6 implants were installed using the palatine bone. The patients’ ages ranged from 40 to 73 years with an average age of 63.5 years. The patients had maxillary sinus-related diseases, such as maxillary sinusitis, oroantral fistula, and dentigerous cysts, prior to implantation. To achieve initial stability, the implants were placed on the palatal side, and buccally tilted. The average postoperative recovery period was 8 months. No postoperative complications occurred in any of the cases, and the approach was used without reported issues. @*Conclusion@#Implant treatment by securing the initial fixation in the palatal bone is a good alternative when an implant must be installed in a patient who requires extensive and invasive bone graft.

Kudoa ogawai (Myxosporea: Kudoidae) Infection in Cultured Olive Flounder Paralichthys olivaceus

Since Kudoa septempuntata was identified as a causative agent of food poisoning associated with raw olive flounder Paralichthys olivaceus, interest and concern regarding the parasite have increased. However, there have been no investigations or reports of other Kudoa species infecting the fish (except for K. paralichthys, which infects the brain) in Korea. We found cysts filled with myxospores of Kudoa species in muscles of cultured olive flounder specimens and identified these to the species level. Mature spores were quadrate, measuring 8.7±0.5 μm in length, 9.2±0.4 μm in thickness, and 12.9±0.6 μm in width. The spores containing 4 polar capsules had a length of 2.1±0.2 μm and a width of 1.8±0.3 μm. The partial 18S and 28S rDNA of isolates showed 99–100% similarities with K. ogawai. Using these morphological and molecular analyses, the species was identified as K. ogawai. This study is the first report of K. ogawai infection in cultured olive flounder in Korea.

Sinuolinea capsularis (Myxosporea: Sinuolineidae) Isolated from Urinary Bladder of Cultured Olive Flounder Paralichthys olivaceus

Sinuolinea capsularis Davis, 1917 is myxosporean that infect the urinary system of the host fish. Insufficient morphological and molecular data of S. capsularis exits, and it is therefore difficult to make an accurate identification of the parasite. We tried a series of morphological and molecular analysis to identify an myxosporean isolated from urinary bladder of cultured olive flounder, Paralichthys olivaceus, from Jeju island in the Republic of Korea. Some of them were observed under a light microscope and SEM, and remain samples were used molecular and phylogenetic analysis. Mature spores were subspherical, measuring 13.9±0.6 μm in length and 13.8±0.8 μm in width. Two spherical polar capsules on opposite sides in the middle of the spore had a diameter range of 4.3±0.4 μm. Scanning electron microscopy revealed that spores a severely twisted the suture line. By the morphological comparison and analysis, it was identified as S. capsularis. In addition, we obtained the partial 18S rDNA of S. capsularis and first registered it in NCBI. Phylogenetic analysis showed that S. capsularis clustered with Zschokkella subclade infecting the urinary system of marine fish, and it supported the infection site tropism effect on phylogeny of marine myxosporeans as well as the origin of Sinuolinea is not monophyly.

Influence of Atlantoaxial Fusion on Sagittal Alignment of the Occipitocervical and Subaxial spines in Os Odontoideum with Atlantoaxial Instability

STUDY DESIGN: Retrospective case analysis. PURPOSE: We hypothesized that larger the C1–C2 fusion angle, greater the severity of the sagittal malalignment of C0–C1 and C2–C7. OVERVIEW OF LITERATURE: In our experience, instances of sagittal malalignment occur at C0–C1 and C2–C7 following atlantoaxial fusion in patients with Os odontoideum (OO). METHODS: We assessed 21 patients who achieved solid atlantoaxial fusion for reducible atlantoaxial instability secondary to OO. The mean patient age at the time of the operation was 42.8 years, and the mean follow-up duration was 4.9 years. Radiographic parameters were preoperatively measured and at the final follow-up. The patients were divided into two groups (A and B) depending on the C1–C2 fusion angle. In group A (n=11), the C1–C2 fusion angle was ≥22°, whereas in group B, it was <22°. The differences in the radiographic parameters of the two groups were evaluated. RESULTS: At the final follow-up, the C1–C2 angle was increased. However, this increase was not statistically significant (18° vs. 22°, p=0.924). The C0–C1 angle (10° vs. 5°, p<0.05) and C2–C7 angle (22° vs. 13°, p<0.05) significantly decreased. The final C1–C2 angle was negatively correlated with the final C0–C1 and C2–C7 angles. The final C0–C1 angle (4° vs. 6°, p<0.05) and C2–C7 angle (8° vs. 20°, p<0.05) were smaller in group A than in group B. After atlantoaxial fusion, the C0–C1 range of motion (ROM; 17° vs. 9°, p<0.05) and the C2–C7 ROM (39° vs. 31°, p<0.05) were significantly decreased. CONCLUSIONS: We found a negative association between the sagittal alignment of C0–C1 and C2–C7 after atlantoaxial fusion and the C1–C2 fusion angle along with decreased ROM. Therefore, overcorrection of C1–C2 kyphosis should be avoided to maintain good physiologic cervical sagittal alignment.

Posterior Sublaminar Wiring and/or Transarticular Screw Fixation for Reducible Atlantoaxial Instability Secondary to Symptomatic Os Odontoideum: A Neglected Technique?

STUDY DESIGN: Retrospective case analysis. PURPOSE: We retrospectively evaluated the clinical and radiological outcomes of posterior sublaminar wiring (PSLW) and/or transarticular screw fixation (TASF) for reducible atlantoaxial instability (AAI) secondary to os odontoideum. OVERVIEW OF LITERATURE: Limited information is available about the surgical outcomes of symptomatic os odontoideum with AAI. METHODS: We examined 23 patients (12 women and 11 men) with os odontoideum and reducible AAI. The average age of the patients at the time of the operation was 44.2 years. The average follow-up duration was 4.5 years. Thirteen patients with anterior AAI underwent PSLW alone, while 10 patients with combined (anterior+posterior) AAI underwent PSLW and TASF. An autogenous iliac bone graft was used for all patients. Nine patients complained of neck or suboccipital pain, and 14 complained of myelopathy. RESULTS: Angulational instability (preoperative 18.7°±8.9° vs. postoperative 2.1°±4.6°, p<0.001), translational instability (16.3±4.9 mm vs. 1.8±2.2 mm, p<0.001), and segmental angle of the C1–C2 joint (23.7°±7.2° vs. 28.4°±3.8°, p<0.05) showed significant improvement postoperatively. Neck Visual Analog Scale score (6.2±2.4 vs. 2.5±1.8, p<0.05) and the modified Japanese Orthopedic Association (9.1±3.1 vs. 13.2±2.6, p<0.05) score also improved, with a recovery rate of 51.8%. Among the three patients who developed nonunion and/or wire breakage, one underwent revision surgery with repeat PSLW and was finally able to achieve fusion. The final fusion rate was 91.3%. CONCLUSIONS: PSLW and/or TASF provided satisfactory clinical and radiological outcomes in reducible AAI secondary to os odontoideum without significant neurological complications. Our results suggest that PSLW and/or TASF can be considered a viable surgical option over segmental fixation in highly selected cases of os odontoideum with reducible AAI.

Mitochondria-targeting drug conjugates for cytotoxic, anti-oxidizing and sensing purposes: current strategies and future perspectives

Mitochondrial targeting is a promising approach for solving current issues in clinical application of chemotherapy and diagnosis of several disorders. Here, we discuss direct conjugation of mitochondrial-targeting moieties to anticancer drugs, antioxidants and sensor molecules. Among them, the most widely applied mitochondrial targeting moiety is triphenylphosphonium (TPP), which is a delocalized cationic lipid that readily accumulates and penetrates through the mitochondrial membrane due to the highly negative mitochondrial membrane potential. Other moieties, including short peptides, dequalinium, guanidine, rhodamine, and F16, are also known to be promising mitochondrial targeting agents. Direct conjugation of mitochondrial targeting moieties to anticancer drugs, antioxidants and sensors results in increased cytotoxicity, anti-oxidizing activity and sensing activity, respectively, compared with their non-targeting counterparts, especially in drug-resistant cells. Although many mitochondria-targeted anticancer drug conjugates have been investigated and , further clinical studies are still needed. On the other hand, several mitochondria-targeting antioxidants have been analyzed in clinical phases I, II and III trials, and one conjugate has been approved for treating eye disease in Russia. There are numerous ongoing studies of mitochondria-targeted sensors.

In vitro characterization of jellyfish venom fibrin(ogen)olytic enzymes from Nemopilema nomurai

Background: Because jellyfish are capable of provoking envenomation in humans, they are considered hazardous organisms. Although the effects of their toxins are a matter of concern, information on the venom components, biological activity and pathological mechanisms are still scarce. Therefore, the aim of the present study was to investigate a serine protease component of Nemopilema nomurai jellyfish venom (NnV) and unveil its characteristics. Methods: To determine the relationship between fibrinolytic activity of NnV and the serine protease, fibrin zymography was performed using metalloprotease and serine protease inhibitors. The biochemical characterization of serine proteases of NnV were determined by the amidolytic assay. Fractions with fibrinolytic activity were obtained by DEAE cation exchange column. Results: NnV displayed fibrinolytic activities with molecular masses of approximately 70, 35, 30, and 28 kDa. The fibrinolytic activity of NnV was completely obliterated by phenylmethylsulfonyl fluoride, a prototype serine protease inhibitor. Based on amidolytic assays using chromogenic substrates specific for various kinds of serine proteases, NnV predominantly manifested a chymotrypsin-like feature. Its activity was completely eliminated at low pH (< 6) and high temperatures (> 37 °C). Some metal ions (Co2+, Cu2+, Zn2+ and Ni2+) strongly suppressed its fibrinolytic activity, while others (Ca2+ and Mg2+) failed to do so. Isolation of a serine protease with fibrionolytic activity from NnV revealed that only p3 showed the fibrinolytic activity, which was completely inhibited by PMSF. Conclusion: The present study showed that N. nomurai jellyfish venom has a chymotrypsin-like serine protease with fibrinolytic activity. Such information might be useful for developing clinical management of jellyfish envenomation and pharmacological agents with therapeutic potential for thrombotic diseases in the future.(AU)

Progress on graft and fixation options of arthroscopic anterior cruciate ligament reconstruction / 中国骨伤

Anterior cruciate ligament injury is one of the most common injuries of knee joint. Arthroscopic reconstruction of anterior cruciate ligament is the most commonly performed procedure. A variety type of fixation techniques are being used nowadays, such as interference screw, cortical suspension fixation and transfix, all of which can achieve good effect, with both advantages and disadvantages. However, the healing of reconstructed grafts is a complicated and long-term process, which can be affected by many factors. The mechanical properties of the grafts are also changed greatly in all stages of healing process, adding variables for the post-operative rehabilitation.

In vitro characterization of jellyfish venom fibrin(ogen)olytic enzymes from Nemopilema nomurai

Abstract Background: Because jellyfish are capable of provoking envenomation in humans, they are considered hazardous organisms. Although the effects of their toxins are a matter of concern, information on the venom components, biological activity and pathological mechanisms are still scarce. Therefore, the aim of the present study was to investigate a serine protease component of Nemopilema nomurai jellyfish venom (NnV) and unveil its characteristics. Methods: To determine the relationship between fibrinolytic activity of NnV and the serine protease, fibrin zymography was performed using metalloprotease and serine protease inhibitors. The biochemical characterization of serine proteases of NnV were determined by the amidolytic assay. Fractions with fibrinolytic activity were obtained by DEAE cation exchange column. Results: NnV displayed fibrinolytic activities with molecular masses of approximately 70, 35, 30, and 28 kDa. The fibrinolytic activity of NnV was completely obliterated by phenylmethylsulfonyl fluoride, a prototype serine protease inhibitor. Based on amidolytic assays using chromogenic substrates specific for various kinds of serine proteases, NnV predominantly manifested a chymotrypsin-like feature. Its activity was completely eliminated at low pH ( 6) and high temperatures (> 37 °C). Some metal ions (Co2+, Cu2+, Zn2+ and Ni2+) strongly suppressed its fibrinolytic activity, while others (Ca2+ and Mg2+) failed to do so. Isolation of a serine protease with fibrionolytic activity from NnV revealed that only p3 showed the fibrinolytic activity, which was completely inhibited by PMSF. Conclusion: The present study showed that N. nomurai jellyfish venom has a chymotrypsin-like serine protease with fibrinolytic activity. Such information might be useful for developing clinical management of jellyfish envenomation and pharmacological agents with therapeutic potential for thrombotic diseases in the future.

Anterior Cervical Fusion Using a Zero-Profile Stand-Alone Cage: Radiological and Clinical Outcomes after More than 2 Years of Follow-Up

STUDY DESIGN: Retrospective analysis. OBJECTIVES: To analyze the usefulness of Zero-P® by analyzing the radiological and clinical outcomes with a minimum of 2 years of follow-up. SUMMARY OF LITERATURE REVIEW: Anterior discectomy and fusion (ACDF) using Zero-P® showed excellent results. However, there is a lack of studies focusing on long-term outcomes. MATERIALS AND METHODS: Ninety-eight patients who underwent single-level ACDF using Zero-P® with more than 2 years of follow-up were included for analysis. In the radiological analysis, we evaluated disc height, segmental lordosis, and subsidence in the preoperative, postoperative, and last follow-up periods. The Neck Disability Index (NDI), Neck Visual Analogue Scale (VAS), and Arm VAS were also evaluated for clinical assessment. Radiological bony fusion was assessed, and radiological and clinical differences according to bony fusion were evaluated. RESULTS: Intervertebral disc height had increased 2.92 mm after surgery and subsided to 2.36 mm at the final follow-up. Subsidence of the screw was 0.58 mm at final follow-up. Segmental lordosis was 3.97° at the preoperative assessment, 8.39° in the postoperative follow-up, and 5.83° in the last follow-up. The Neck VAS score was 4.47, 2.28, and 1.27, respectively. The Arm VAS score was 5.73, 3.13, and 2.18; and NDI score was 17.8, 11.7, and 7.89, respectively. There was no association between the radiological and clinical results. Radiological nonunion was found in 18 subjects. There were no significant differences in radiological and clinical parameters according to bony union. CONCLUSIONS: ACDF with Zero-P® for treatment of degenerative cervical disease showed subsidence in 55.1% and nonunion in 18% of cases. However, the radiological results were not related to the clinical results.