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Purpose: Despite the clinical utility of the pivot shift exam, the requisite applied forces and torques to elicit a pivot shift remain unclear. The purposes of this study are (1) to identify the greatest forces and torques applied to the knee during the pivot shift exam and (2) to evaluate if the applied loads differ among experienced surgeons., Methods: Three cadaveric hemipelvis-to-toe specimens (ages 53, 36 and 31 years; two males and one female) with no history of knee or hip injury were utilized. The experimental setup consisted of securing the hemipelvis to a mounting frame via an external fixator to simulate patient positioning during the clinical exam. The hemipelvis, femur, and tibia were spatially tracked by motion capture and the applied loads were measured using a 6-axis force-torque sensor. After sectioning the anterior cruciate ligament (ACL), three board-certified sports medicine surgeons then performed the pivot shift exam on each specimen utilizing their preferred technique. Forces (compression-distraction, anterior-posterior, and medial-lateral) and torques (varus-valgus, internal-external rotation, and flexion-extension) applied to the knee joint immediately preceding the reduction of the proximal lateral tibia during each pivot shift exam were calculated., Results: Compression was the largest applied force averaging 95 N ± 15 N for all surgeons and knees, which was at least 4.5 times greater, on average, than the applied anterior and applied medial tibial forces (p < 0.0001). Valgus was the largest of the three applied torques, averaging 8.5 ± 2.1 Nm. Internal rotation torque was 3.7 times less, on average, than the applied valgus torque (p < 0.0001). Each surgeon applied compressive force. However, anterior force was more variable among surgeons, with one of the three surgeons applying minimal anterior force (p ≤ 0.024). The magnitude of applied torques was similar among examiners (n.s)., Conclusion: Compressive force and valgus torque were the predominant applied loads during the pivot shift exam. A lower magnitude of internal rotation torque was also applied. The anterior force was not consistently applied among examiners. These data can better inform clinical, cadaveric, and computational studies utilizing the pivot shift exam to assess knee biomechanics and can be used to educate trainees in conducting this complex manoeuvre., Level of Evidence: An in vitro biomechanic study., (© 2024 European Society of Sports Traumatology, Knee Surgery and Arthroscopy.)

Physical examination of the knee joint is used to diagnose the type and severity of knee ligament injury; however, these exams are qualitative and subjective. To perform common physical examinations, we developed an arthrometer which quantitatively measures the load-displacement response in anterior-posterior (AP) translation, internal-external rotation (IER) and varus-valgus (VV) rotation. Here we describe safety, reliability, minimum detectable changes (MDCs), and absolute side-to-side differences in twenty young, healthy subjects (ten male, ten female, mean age: 28 ± 6 years). The arthrometer consists of an instrumented mechanical linkage, a force-moment sensor, and software for real-time visualization and recording of the load-displacement responses. During testing, the subject sits reclined in a chair with their knee fixed at 30° of flexion. Two examiners tested both knees of each subject twice to assess reliability via intraclass correlation coefficients (ICC). All subjects completed the test protocol with minimal pain and stated that they would volunteer to be tested again. Each knee required on average five minutes to test. All intra-test reliabilities were excellent (≥0.91). Intra-examiner reliabilities ranged from good to excellent (0.62-0.89), and inter-examiner reliabilities were good to excellent (≥0.72). MDCs for repeat measures were ≤ 4.5 mm, 4.6°, and 2.3° for AP, IER, and VV, respectively. The absolute side-to-side differences for this cohort averaged 3.8 mm in AP, 5.5° in IER, and 2.2° in VV. Our arthrometer was safe, testing was time-efficient, and MDCs in our cohort of healthy subjects support utilization of this device for clinical research., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Carl W. Imhauser – Received speaker fees for Corin Group. Erin E. Berube, Emmanuel Akinola Oladimeji, David Z. Shamritsky, Zaid A. Zayyad, Thomas J. Fraychineaud, Jennifer T. Vazquez, Hamidreza Jahandar, Stephen Lyman, Debi Jones – have no conflict of interest. Andrew D. Pearle − Received consulting fees from Depuy, Exactech, Medical Device Business Services, Smith & Nephew, Stryker Corporation, and Zimmer Biomet; royalties from Zimmer Biomet; and education support from Arthrex; and has ownership interest in Engage Surgical, Knee Guardian, PerfectFit, MyGemini, and Therma1. Michael Parides – have no conflict of interest. Danyal H. Nawabi − Received education support from Arthrex and Gotham Surgical Solutions; consulting fees from Conmed, Linvatec, and Newclip USA, hospitality payments from Stryker Corporation. Theresa A. Chiaia – have no conflict of interest. Thomas L. Wickiewicz − Received royalties from Stryker Corporation]., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Traumatic brain injury (TBI) is a significant global health concern and is characterized by brain dysfunction resulting from external physical forces, leading to brain pathology and neuropsychiatric disorders such as anxiety. This study investigates the effects of TC-DAPK6 on tau hyper-phosphorylation, gene expression, anxiety, and behavior impairment in the TBI mice model., Methods and Results: A weight drop model induced the TBI and the anxiety levels were evaluated using an elevated plus maze (EPM) test. TC-DAPK6 was intraperitoneally administered one-month post-TBI and continued for two months. The total cis-p-tau ratio in the brain was assessed using western blot and immunofluorescence staining. Molecular analysis was conducted on Aff2, Zkscan16, Kcna1, Pcdhac2, and Pcdhga8 to investigate the function and pathogenic role of TC-DAPK6 in neurological diseases in the cerebral cortex tissues of TBI-model mice, and the results were compared with TC-DAPK6 TBI-treatment group. The anxiety level and phosphorylation of tau protein in the TBI group were significantly increased compared to the sham groups and decreased substantially in the TBI-treatment group after TC-DAPK6 administration; the TBI group mostly spent their time with open arms. TC-DAPK6 decreased the expression level of genes as much as the sham group. Meanwhile, KCNA1 showed the highest fold of changes in the TBI and TBI-treatment groups., Conclusions: The study demonstrates a clear association between cis-p-tau and neuro-related gene expression levels in TBI-induced mice. Targeting these pathways with DAPK1 inhibitors, shows promise for therapeutic interventions in TBI and related neurodegenerative disorders., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Anterior cervical discectomy and fusion (ACDF) is known to elicit adverse biomechanical effects on immediately adjacent segments; however, its impact on the kinematics of the remaining nonadjacent cervical levels has not been understood. This study aimed to explore the biomechanical impact of ACDF on kinematics beyond the immediate fusion site. We hypothesized that compensatory motion following single-level ACDF is not predictably distributed to adjacent segments due to compensation from noncontiguous levels., Methods: Six fresh-frozen cervical spines (C2-T1) underwent fluoroscopic screening and sagittal and coronal reformats from computed tomography scans and were utilized to grade segmental degeneration. Each specimen was tested to 30° of flexion and extension intact and following single-level ACDF at the C5-C6 level. The motions of each vertebral body were tracked using 3-dimensional (3D) motion capture into an inverse kinematics model, facilitating correlations between the 3D reconstruction from computed tomography images and the 3D motion capture data. This model was used to calculate each level's flexion/extension range of motion (ROM)., Results: Single-level fusion at the C5-C6 level across all specimens resulted in a significant motion reduction of -6.8° ( P = 0.002). No significant change in ROM occurred in the immediate adjacent segments C4-C5 ( P = 0.07) or C6-C7 ( P = 0.15). Hypermobility was observed in 2 specimens (33%) exclusively in adjacent segments. In contrast, the other 4 spines (66%) displayed hypermobility at noncontiguous segments. Hypermobility occurred in 42% (5/12) of the adjacent segments, 28% (5/18) of the noncontiguous segments, and 50% (3/6) of the cervicothoracic segments., Conclusion: Single-level ACDF impacts ROM beyond adjacent segments, extending to noncontiguous levels. Compensatory motion, not limited to adjacent levels, may be influenced by degenerative changes in noncontiguous segments. Surprisingly, hypermobility may not occur in adjacent segments after ACDF., Clinical Relevance: Overall, the multifaceted biomechanical effects of ACDF underscore the need for a comprehensive understanding of cervical spine dynamics beyond immediate adjacency, and it needs to be taken into consideration when planning single-level ACDF., Competing Interests: Declaration of Conflicting Interests : The authors report no conflicts of interest in this work., (This manuscript is generously published free of charge by ISASS, the International Society for the Advancement of Spine Surgery. Copyright © 2024 ISASS. To see more or order reprints or permissions, see http://ijssurgery.com.)

From the environmental point of view, azo dye industrial effluent is a major public health concern due to its toxic, carcinogenic, and teratogenic characteristics. On the other hand, using enzyme-based technologies offers a promising systematic and controllable method for removing synthetic dyes from wastewater. In the present study, yttrium (Y 3+ ) phosphate was applied for the synthesis of hybrid nanoparticles (HNPs) consisting of laccase as the green catalyst. When the association of HNPs was fixed by glutaraldehyde (GA), three-dimensional cubic structures with the regular arrangement were provided. GA increased the reusability of the fabricated hybrid nanostructures (HNSs) up to 32 successive cycles. About 85% of Direct Blue-15 was removed after a 4 h-treatment using laccase@YPO 4 •HNPs and laccase@GA@YPO 4 •HNSs. The azo dye removal data were well-fitted with a pseudo-second-order model for both types of the prepared HNSs. For the model freshwater green alga Raphidocelis subcapitata, the half maximal effective concentration (EC 50 ) of the dye decreased 10- and 100-fold after the removal with laccase@YPO 4 •HNPs and laccase@GA@YPO 4 •HNSs, respectively. GA-treated HNSs (250 U L -1 ) inhibited the biofilm formation by approximately 78%, 82%, and 79% for Escherichia coli, Staphylococcus aureus, and Bacillus subtilis, respectively. Thus, the fabricated laccase@GA@YPO 4 •HNSs could be presented as a novel, efficient, and recyclable heterogeneous biocatalyst for wastewater treatment and clean-up., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

High-grade knee laxity is associated with early anterior cruciate ligament (ACL) graft failure, poor function, and compromised clinical outcome. Yet, the specific ligaments and ligament properties driving knee laxity remain poorly understood. We described a Bayesian calibration methodology for predicting unknown ligament properties in a computational knee model. Then, we applied the method to estimate unknown ligament properties with uncertainty bounds using tibiofemoral kinematics and ACL force measurements from two cadaver knees that spanned a range of laxities; these knees were tested using a robotic manipulator. The unknown ligament properties were from the Bayesian set of plausible ligament properties, as specified by their posterior distribution. Finally, we developed a calibrated predictor of tibiofemoral kinematics and ACL force with their own uncertainty bounds. The calibrated predictor was developed by first collecting the posterior draws of the kinematics and ACL force that are induced by the posterior draws of the ligament properties and model parameters. Bayesian calibration identified unique ligament slack lengths for the two knee models and produced ACL force and kinematic predictions that were closer to the corresponding in vitro measurement than those from a standard optimization technique. This Bayesian framework quantifies uncertainty in both ligament properties and model outputs; an important step towards developing subject-specific computational models to improve treatment for ACL injury., (Copyright © 2023 by ASME.)

Aims: Mid-level constraint designs for total knee arthroplasty (TKA) are intended to reduce coronal plane laxity. Our aims were to compare kinematics and ligament forces of the Zimmer Biomet Persona posterior-stabilized (PS) and mid-level designs in the coronal, sagittal, and axial planes under loads simulating clinical exams of the knee in a cadaver model., Methods: We performed TKA on eight cadaveric knees and loaded them using a robotic manipulator. We tested both PS and mid-level designs under loads simulating clinical exams via applied varus and valgus moments, internal-external (IE) rotation moments, and anteroposterior forces at 0°, 30°, and 90° of flexion. We measured the resulting tibiofemoral angulations and translations. We also quantified the forces carried by the medial and lateral collateral ligaments (MCL/LCL) via serial sectioning of these structures and use of the principle of superposition., Results: Mid-level inserts reduced varus angulations compared to PS inserts by a median of 0.4°, 0.9°, and 1.5° at 0°, 30°, and 90° of flexion, respectively, and reduced valgus angulations by a median of 0.3°, 1.0°, and 1.2° (p ≤ 0.027 for all comparisons). Mid-level inserts reduced net IE rotations by a median of 5.6°, 14.7°, and 17.5° at 0°, 30°, and 90°, respectively (p = 0.012). Mid-level inserts reduced anterior tibial translation only at 90° of flexion by a median of 3.0 millimetres (p = 0.036). With an applied varus moment, the mid-level insert decreased LCL force compared to the PS insert at all three flexion angles that were tested (p ≤ 0.036). In contrast, with a valgus moment the mid-level insert did not reduce MCL force. With an applied internal rotation moment, the mid-level insert decreased LCL force at 30° and 90° by a median of 25.7 N and 31.7 N, respectively (p = 0.017 and p = 0.012). With an external rotation moment, the mid-level insert decreased MCL force at 30° and 90° by a median of 45.7 N and 20.0 N, respectively (p ≤ 0.017 for all comparisons). With an applied anterior load, MCL and LCL forces showed no differences between the two inserts at 30° and 90° of flexion., Conclusion: The mid-level insert used in this study decreased coronal and axial plane laxities compared to the PS insert, but its stabilizing benefit in the sagittal plane was limited. Both mid-level and PS inserts depended on the MCL to resist anterior loads during a simulated clinical exam of anterior laxity., Competing Interests: M. B. Cross reports royalties or licenses from Depuy Synthes and PSI, consulting fees from Depuy Synthes, Exactech, and 3M, payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing or educational events from Depuy Synthes and 3M, and stock or stock options in Ospitek, PSI, and Intellijoint, all of which are unrelated to this article. D. J. Mayman reports royalties or licenses from Orthalign and Smith & Nephew, and stock or stock options in Imagen, Orthalign, Smith & Nephew, Wishbone, Micare Path, and Cymedica, all of which are unrelated to this article. C. W. Imhauser reports payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing or educational events from Corin Medical, unrelated to this study. P. K. Sculco reports consulting fees from Zimmer Biomet, Intellijoint Surgical, Lima Corporate, Depuy Synthes, and ATEC, payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing or educational events from Zimmer Biomet, stock or stock options in Parvizi Surgical Innovation and Intellijoint Surgical, and receipt of equipment, materials, drugs, medical writing, gifts or other services, all of which are unrelated to this study., (© 2023 Author(s) et al.)

Objective: One of the main approaches to preventing skin ageing is to protect fibroblast cells from oxidative stress. The promoting effect of the human amniotic membrane extract (hAME) on re-epithelization, proliferation and migration of cells in wound healing has been already well studied. This experimental study aimed to investigate the antioxidant activity of hAME against hydrogen peroxide (H 2 O 2 )-induced dermal fibroblast damage., Methods: Here, to establish the ageing model, human foreskin fibroblasts (HFFs) were exposed to 200 μM H 2 O 2 for 2 h. HFFs were treated with 0.1 mg/ml AME for 24 or 48 h before or/and after H 2 O 2 exposure. A total of 48 h following the H 2 O 2 treatment, we measured cell proliferation, viability, senescence-associated β-galactosidase (SA-β-Gal), antioxidants and preinflammatory cytokine (IL-6) levels, as well as the expression of senescence-associated genes (P53 and P21)., Results: The obtained results indicated that under oxidative stress, AME significantly increased cellular viability and not only promoted the cell proliferation rate but also attenuated apoptotic induction condition (p < 0.001). AME also significantly reversed the SA-β-Gal levels induced by H 2 O 2 (p < 0.001). Additionally, both pre- and post-treatment regimen by AME down-regulated the expression of senescence marker genes (p < 0.001). Moreover, AME declined different oxidative stress biomarkers such as superoxide dismutase and catalase and increased the glutathione amount., Conclusion: Altogether, our results indicated that AME had a remarkable antioxidant and antiageing activity as pre- and post-treatment regimen, pointing to this compound as a potential natural-based cosmeceutical agent to prevent and treat skin ageing conditions., (© 2022 Society of Cosmetic Scientists and Societe Francaise de Cosmetologie.)

Aims & Background: Pilocarpine is an accepted treatment for xerostomia, but limited research has been conducted on the oral, topical form. The present study aimed to compare the effects of 1 and 2% pilocarpine mouthwash on xerostomic participants., Methods: In this double-blind clinical trial study, 48 subjects with xerostomia were randomly divided into three groups to measure the effects of 1 and 2% pilocarpine and placebo mouthwashes on saliva levels. The amount of saliva in the 1st and 14th days was measured at 0, 45, 60, and 75 mins, while participants used their mouthwash three times a day for 14 days. On the 1st and 14th days, they filled out the information forms on xerostomia and the medicine's side effects before and after the intervention., Results: On the 1st day, the mean salivary flow at 45, 60, and 75 mins in the 2 and 1% pilocarpine mouthwash were significantly higher than in the placebo mouthwash group (p < 0.05). On the 14th day, the mean salivary flow time at 45 mins in the 2% pilocarpine mouthwash group was significantly higher than in the placebo mouthwash group (p = 0.007). Furthermore, the mean salivary flow at 60 and 75 mins in the 2% (p < 0.001) and 1% pilocarpine mouthwash (p = 0.028) was significantly higher than in the placebo group. Moreover, the salivary flow in the 2% pilocarpine mouthwash group was significantly higher than the 1% pilocarpine mouthwash (p < 0.05) during these two times. No side effects were observed in any of the subjects., Conclusions: The study showed that 5 ml of 2 and 1% pilocarpine mouthwash for 2 weeks increased salivary flow in xerostomic participants compared to placebo without any side effects., (© 2022. The Author(s).)

Laccase was successfully hybridized with polyamidoamine (PAMAM) dendrimer-grafted silica-coated hercynite-copper phosphate magnetic hybrid nanoflowers (MHNFs) to increase the catalytic performance of the enzyme and apply in an effective bioremoval of gemifloxacin. For this purpose, the magnetic nanoparticles (MNPs) of hercynite were covered with a silica layer, and the core-shell SiO 2 @hercynite was then modified with PAMAM dendrimer to increase the surface area of the carrier for the enzyme attachment. Subsequently, the whole complex was hybridized with laccase and copper phosphate to attain a large surface area (104.3 m 2  g -1 ). The fabricated MHNFs acquired the entrapment yield and efficiency of 90 ± 3% and 66 ± 5%, respectively. The catalytic activity of the fabricated biocatalyst was remained up to 50% after 13 reusability cycles. Approximately 90% of gemifloxacin was removed by the constructed MHNFs after 3 h incubation by adsorption and degradation mechanisms. The biotransformation products were then identified, and degradation pathways were proposed as defluorination, decarboxylation, elimination of a cyclopropyl group, and cleavage of the pyrrolidine moiety. Furthermore, the toxicity of gemifloxacin was effectively diminished against some bacterial strains., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

An efficient heterogeneous natural polymer-based biocatalyst was fabricated through the immobilization of laccase onto dialdehyde inulin (DAI)-coated silica-caped magnetic nanoparticles (laccase@DAI@SiO 2 @Fe 3 O 4 ⋅MNPs). The carrier was developed using SiO 2 @Fe 3 O 4 ⋅MNPs and functionalized with DAI. The construction of immobilized laccase was confirmed by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy. Immobilization yield and efficiency were calculated as 61.0 ± 0.3% and 93.0 ± 0.6%, respectively. The immobilized laccase maintained 50% and 85% of its relative activity after 25 repeated cycles and 20 days of storage at 4 °C, respectively. The prepared biocatalyst effectively eliminated ofloxacin, a fluoroquinolone-type antibiotic, with a 63% removal capacity. Besides, antimicrobial activity study on some soil microorganisms involved in the biodegradation of xenobiotics revealed that the laccase-treated ofloxacin resulted in less toxic metabolites. The obtained data indicated that the fabricated biocatalyst is promising for the removal of ofloxacin or other analogs of fluoroquinolones in the environment., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

A novel approach termed the "concentrated method" was developed for the instant fabrication of laccase@Co 3 (PO 4 ) 2 •hybrid nanoflowers (HNFs). The constructed HNFs were obtained by optimizing the concentration of cobalt chloride and phosphate buffer to reach the highest activity recovery. The incorporation of 30 mM CoCl 2 and 160 mM phosphate buffer (pH 7.4) resulted in a fast anisotropic growth of the nanomaterials. The purposed method did not involve harsh conditions and prolonged incubation of precursors, as the most reported approaches for the synthesis of HNFs. The catalytic efficiency of the immobilized and free laccase was 460 and 400 M -1 S -1 , respectively. Also, the enzymatic activity of the prepared biocatalyst was 113% of the free enzyme (0.5 U mL -1 ). The stability of the synthesized HNFs was enhanced by 400% at pH 6.5-9.5 and the elevated temperatures. The activity of laccase@Co 3 (PO 4 ) 2 •HNFs declined to 50% of the initial value after 10 reusability cycles, indicating successful immobilization of the enzyme. Structural studies revealed a 32% increase in the α-helix content after hybridization with cobalt phosphate, which improved the activity and stability of the immobilized laccase. Furthermore, the fabricated HNFs exhibited a considerable ability to remove moxifloxacin as an emerging pollutant. The antibiotic (10 mg L -1 ) was removed by 24% and 75% after 24 h through adsorption and biodegradation, respectively. This study introduces a new method for synthesizing HNFs, which could be used for the fabrication of efficient biocatalysts, biosensors, and adsorbents for industrial, biomedical, and environmental applications., (© 2022. The Author(s).)

Objectives: Metformin (Met) and liraglutide (Lira) have been approved to treat type 2 diabetes mellitus and have cardioprotective effects., Materials and Methods: Human umbilical vein endothelial cells (HUVECs) were incubated with Met, Lira, or their combination in this study., Results: Results showed that the synergistic inhibitory effect of the two drugs on HUVECs proliferation was significant (75%) after 48 hr drug exposure. In addition, either Lira or Met alone had a marked tendency to inhibit the migration of HUVECs (42% and 39%). Almost a complete inhibition (97%) was demonstrated in combinational use after 48 hr treatment. After combining these two drugs, the apoptosis rate raised to 68%, which was a significant approval of synergistic apoptosis induction of Met and Lira. The combinational group indicated a substantial increase in VEGF, PDGF, and MMP-9 at 24 hr compared with the control., Conclusion: This study showed that combination therapy with Lira and Met could effectively reduce cell proliferation, induce apoptosis, and inhibit cell migration in the HUVECs. This study provides evidence to support using Met in combination with Lira as a treatment option for patients with type-2 diabetes and cancer., Competing Interests: The authors declare that there are no conflict of interests regarding this publication.

Background: Due to the association of hypermutated colorectal cancer (CRC) with many neo-antigens, poly-neo-epitopes are attractive vaccines. The molecular features of murine CT26 are similar to those of aggressive human CRC. CT26 contains some antigenic mutations, which can provide specific immunotherapy targets. Herein, we aimed to express, and purify the previously designed hexatope containing CT26 neoepitopes, CT26-poly-neoepitopes., Methods and Results: In the current study, expression of the CT26-poly-neoepitopes was optimized in three different Escherichia coli strains including BL21 (DE3), Origami (DE3), and SHuffle®. Furthermore, the effect of ethanol on the CT26-poly-neoepitopes expression was investigated. The highest amount of CT26-poly-neoepitopes, which included CT26-poly-neoepitopes with the uncleaved pelB signal sequence and the processed one, was achieved when BL21 containing pET-22 (CT26-poly-neoepitopes) was induced with 0.1 mM IPTG for 48 h at 22 ºC in the presence of 2% ethanol. However, 37 ºC was the optimized induction temperature for expression of the CT26-poly-neoepitopes in the absence of ethanol. To purify the CT26-poly-neoepitopes, Ni-NTA affinity chromatography under denaturing and hybrid conditions were applied. High and satisfactory CT26-poly-neoepitopes purity was achieved by the combined urea and imidazole method., Conclusion: The effect of ethanol on expression of the CT26-poly-neoepitopes was temperature-dependent. Furthermore, the pelB-mediated translocation of the CT26-poly-neoepitopes into the periplasm was inefficient. Moreover, higher concentration of imidazole in the washing buffer improved the CT26-poly-neoepitopes purification under hybrid condition. Overall, the immunogenicity of CT26-poly-neoepitopes expressed in BL21 under the optimum condition and purified under hybrid condition can be studied in our future in vivo study., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: There is concern that utilization of lateral extra-articular tenodesis (LET) in conjunction with anterior cruciate ligament (ACL) reconstruction (ACLR) may disturb lateral compartment contact mechanics and contribute to joint degeneration., Hypothesis: ACLR augmented with LET will alter lateral compartment contact mechanics in response to simulated pivoting maneuvers., Study Design: Controlled laboratory study., Methods: Loads simulating a pivot shift were applied to 7 cadaveric knees (4 male; mean age, 39 ± 12 years; range, 28-54 years) using a robotic manipulator. Each knee was tested with the ACL intact, sectioned, reconstructed (via patellar tendon autograft), and, finally, after augmenting ACLR with LET (using a modified Lemaire technique) in the presence of a sectioned anterolateral ligament and Kaplan fibers. Lateral compartment contact mechanics were measured using a contact stress transducer. Outcome measures were anteroposterior location of the center of contact stress (CCS), contact force from anterior to posterior, and peak and mean contact stress., Results: On average, augmenting ACLR with LET shifted the lateral compartment CCS anteriorly compared with the intact knee and compared with ACLR in isolation by a maximum of 5.4 ± 2.3 mm ( P < .001) and 6.0 ± 2.6 mm ( P < .001), respectively. ACLR augmented with LET also increased contact force anteriorly on the lateral tibial plateau compared with the intact knee and compared with isolated ACLR by a maximum of 12 ± 6 N ( P = .001) and 17 ± 10 N ( P = .002), respectively. Compared with ACLR in isolation, ACLR augmented with LET increased peak and mean lateral compartment contact stress by 0.7 ± 0.5 MPa ( P = .005) and by 0.17 ± 0.12 ( P = .006), respectively, at 15° of flexion., Conclusion: Under simulated pivoting loads, adding LET to ACLR anteriorized the CCS on the lateral tibial plateau, thereby increasing contact force anteriorly. Compared with ACLR in isolation, ACLR augmented with LET increased peak and mean lateral compartment contact stress at 15° of flexion., Clinical Relevance: The clinical and biological effect of increased anterior loading of the lateral compartment after LET merits further investigation. The ability of LET to anteriorize contact stress on the lateral compartment may be useful in knees with passive anterior subluxation of the lateral tibia.

Lipase-catalyzed esterification is an efficient technique in the production of polyunsaturated fatty acid (PUFA) concentrates which are applied for nutrition and health purposes. In this project, a solvent-tolerant lipase from Streptomyces pratensis MV1 was immobilized and purified by a hydrophobic support. The purified lipase revealed enhanced activity and stability towards chemicals, organic solvents, and a broad range of pH values. The production of lipase was enhanced to 7.0 U/mL after optimization by a central composite design. Acylglycerols (AGs) rich in α-linolenic acid (45%, w/w) were produced and a favorable n-6/n-3 free fatty acid (FFA) ratio of 1.1 was achieved in fenugreek seed oil using the immobilized lipase. The ability of S. pratensis lipase in ester synthesis and the improvement of n6/n3 FFA ratio make it a suitable candidate in food production industries., (© Association of Food Scientists & Technologists (India) 2020.)

Pseudomonas aeruginosa (PA) is a leading cause of nosocomial infections and death in cystic fibrosis patients. The study was conducted to evaluate the physicochemical structure, biological activity and serum stability of a recombinant anti-PcrV single chain variable antibody fragment genetically attached to the mCH3cc domain. The stereochemical properties of scFv-mCH3 (YFL001) and scFv (YFL002) proteins as well as molecular interactions towards Pseudomonas aeruginosa PcrV were evaluated computationally. The subcloned fragments encoding YFL001 and YFL002 in pET28a were expressed within the E. coli BL21-DE3 strain. After Ni-NTA affinity chromatography, the biological activity of the proteins in inhibition of PA induced hemolysis as well as cellular cytotoxicity was assessed. In silico analysis revealed the satisfactory stereochemical quality of the models as well as common residues in their interface with PcrV. The structural differences of proteins through circular dichroism spectroscopy were confirmed by NMR analysis. Both proteins indicated inhibition of ExoU positive PA strains in hemolysis of red blood cells compared to ExoU negative strains as well as cytotoxicity effect on lung epithelial cells. The ELISA test showed the longer serum stability of the YFL001 molecule than YFL002. The results were encouraging to further evaluation of these two scFv molecules in animal models.

Anti-TNF inhibitors exert their therapeutic effect by inhibition of the excessive amounts of TNF-α within the body. Recombinant TNF-α should be produced in a soluble refolded form to investigate the effectiveness and efficiency of anti-TNF-α compounds. In this research, the designed cassette was subcloned in the pET28a expression vector and expressed in E. coli BL21 (DE3). The identity of the protein was confirmed through SDS-PAGE and Western blotting. After optimizing expression conditions, protein purification was performed using native Ni-NTA affinity chromatography. The biological activity of the soluble recombinant TNF-α was investigated using MTT assay. Also, the affinity of an anti-TNF-α agent, Altebrel, was investigated against the expressed protein through ELISA. Optimization of TNF-α expression conditions represented that the highest expression could be achieved at 37 °C using 0.5 mM IPTG 6 h post-induction. The recombinant protein represented an inhibitory effect on the L929 murine fibroblast cell line and was successfully detected by Altebrel in ELISA. Binding kinetics were also studied using Cimzia as an anti-TNF-α molecule and 7.2 E -13 M was calculated as the equilibrium dissociation constant value (K D ). The significant expression level of the recombinant protein in the soluble form, its high purity, and assessment of its biological activity showed that the expressed protein could be used in tests of ELISA and MTT to assess the activity of anti-TNF-α agents., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: The biomechanical effect of lateral extra-articular tenodesis (LET) performed in conjunction with anterior cruciate ligament (ACL) reconstruction (ACLR) on load sharing between the ACL graft and the LET and on knee kinematics is not clear., Purpose/hypothesis: The purpose was to quantify the effect of LET on (1) forces carried by both the ACL graft and the LET and (2) tibiofemoral kinematics in response to simulated pivot shift and anterior laxity tests. We hypothesized that LET would decrease forces carried by the ACL graft and anterior tibial translation (ATT) in response to simulated pivoting maneuvers and during simulated tests of anterior laxity., Study Design: Controlled laboratory study., Methods: Seven cadaveric knees (mean age, 39 ± 12 years [range, 28-54 years]; 4 male) were mounted to a robotic manipulator. The robot simulated clinical pivoting maneuvers and tests of anterior laxity: namely, the Lachman and anterior drawer tests. Each knee was assessed in the following states: ACL intact, ACL sectioned, ACL reconstructed (using a bone-patellar tendon-bone autograft), and after performing LET (the modified Lemaire technique after sectioning of the anterolateral ligament and Kaplan fibers). Resultant forces carried by the ACL graft and LET at the peak applied loads were determined via superposition. ATT was determined in response to the applied loads., Results: With the applied pivoting loads, performing LET decreased ACL graft force up to 80% (44 ± 12 N; P < .001) and decreased ATT of the lateral compartment compared with that of the intact knee up to 7.6 ± 2.9 mm ( P < .001). The LET carried up to 91% of the force generated in the ACL graft during isolated ACLR (without LET). For simulated tests of anterior laxity, performing LET decreased ACL graft force by 70% (40 ± 20 N; P = .001) for the anterior drawer test with no significant difference detected for the Lachman test. No differences in ATT were deteced between ACLR with LET and the intact knee on both the Lachman and the anterior drawer tests ( P = .409). LET reduced ATT compared with isolated ACLR on the simulated anterior drawer test by 2.4 ± 1.8 mm ( P = .032) but not on the simulated Lachman test., Conclusion: In a cadaveric model, LET in combination with ACLR transferred loads from the ACL graft to the LET and reduced ATT with applied pivoting loads and during the simulated anterior drawer test. The effect of LET on ACL graft force and ATT was less pronounced on the simulated Lachman test., Clinical Relevance: LET in addition to ACLR may be a suitable option to offload the ACL graft and to reduce ATT in the lateral compartment to magnitudes less than that of the intact knee with clinical pivoting maneuvers. In contrast, LET did not offload the ACL graft or add to the anterior restraint provided by the ACL graft during the Lachman test.

Bisphenol A (BPA) is an environmental pollutant with adverse effects on different ecosystems. In this study, immobilized laccase enzymes onto inorganic supports were used to remove BPA. Laccase was successfully immobilized on sodium zeolite Y (NaY) and its modified desilicated (DSY) and dealuminated (DAY) forms. NaY-based supports were instrumentally characterized. The immobilized laccase on NaY (laccase@NaY), desilicated (laccase@DSY), and dealuminated (laccase@DAY) forms showed significant improvement on immobilization yield (IY%) and efficiency (IE%). Laccase@DSY and laccase@NaY showed IY% = 73.18 ± 3.33 % and 46.23 ± 1.81 % and IE% = 94.50 ± 1.86 %, and 74.39 ± 1.41 %, respectively, whereas IY% and IE% for laccase@DAY were achieved as 81.12 ± 1.32 % and 98.56 ± 2.93 %, respectively. The supports also increased the enzyme characteristics such as pH-temperature range, catalytic stability, and reusability. K m values were 0.73 ± 0.05, 0.26 ± 0.09, 0.31 ± 0.5, and 1.01 ± 0.03 mM for laccase@NaY, laccase@DAY, laccase@DSY, and the free enzyme, respectively. The enzyme demonstrated higher biodegradation ability of bisphenol A upon immobilization on the supports compared to that of the soluble enzyme. A bio-removal yield of 86.7 % was obtained considering three parameters including amount of laccase@DAY (8 U mg -1 ), concentration of BPA (0.5 mM), and treatment time (1 h) based on response surface methodology (RSM). Biodegradation metabolites (49 ± 5.8 %) and unconverted BPA (14 ± 5.2 %) were analyzed by gas chromatography-mass spectrometry., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interest., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Injury to the lateral ulnar collateral ligament (LUCL) complex of the elbow often results in posterolateral rotatory instability. Although surgical reconstruction of the LUCL is often required, gaps in our understanding of the LUCL complex remain. The purpose of this study was to provide a robust and accurate characterization of the lateral elbow ligamentous complex., Methods: The LUCLs, radial collateral ligaments, and annular ligaments in 10 cadaveric elbows were 3-dimensionally digitized and reconstructed using computed tomography. Surface areas, origin and insertion footprint areas, distances between perceived footprint centers and geometric footprint centroids, distances to key landmarks, and ligament isometry were measured., Results: The mean surface area of the LUCL was 229.3 mm 2 . The mean origin and insertion footprint areas were 26.0 mm 2 and 22.9 mm 2 , respectively. The mean distance between the apparent centers and the geometric centroids of the footprints was 1 mm. The center of the LUCL origin was 10.7 mm distal to the lateral epicondyle and 8.2 mm from the capitellar articular margin. The center of the LUCL insertion was 3.3 mm distal to the apex of the supinator crest. The LUCL showed anisometric properties as elbow flexion increased (P < .001)., Conclusions: The LUCL origin center was 10.7 mm from the lateral epicondyle, whereas the insertion center was 3.3 mm from the apex of the supinator crest. The visually estimated footprint centers were generally within 1 mm of the geometric centroid. These geometries and distances to key landmarks will be informative for surgeons seeking to perform anatomic ligament reconstruction procedures., (Copyright © 2018 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.)

Purpose: The purposes of this study were (1) to evaluate the percentage of gluteus medius and minimus tendon footprint restoration that can be achieved with fixation using single-row repair versus double-row repair and (2) to evaluate the yield load of a repair of the gluteus medius and minimus tendon using single-row versus double-row repair techniques., Methods: Twelve human fresh-frozen cadaveric hip specimens (6 matched pairs, 4 female, mean age 47.5 ± 14.5 years) were tested. Specimens were excluded if they had any prior hip surgery or injury, if any abnormality of the tendon was noted on dissection, or if they had a body mass index <20 or >35 or a T-score <2.0 on dual-energy x-ray absorptiometry scanning. Matched pairs were randomized to receive either double-row repair with 2 standard suture anchors and 2 knotless anchor devices or a single-row repair with suture anchors only. The percentage of the footprint area covered after repair was determined using a computer-assisted digitization algorithm. With a mechanical testing system, each repaired specimen was tested for mechanical strength first with cyclic loading and then load to failure testing., Results: Footprint coverage of the lateral facet was significantly greater for double-row repair (mean 76.6%) compared with single-row repair (mean 50.3%) (P = .03). There was no significant difference between single- and double-row repair for posterior-superior or anterior facet coverage. Mechanical testing showed a higher mean yield load for double-row anchor repair (197.6 ± 61.7 N vs 163.5 ± 35.4 N for single-row repair), but this did not reach statistical significance (P = .15). The predominant mode of failure was suture pullout through the musculotendinous unit (9/12 specimens: 5 double-row and 4 single-row)., Conclusions: For hip abductor tears, double-row suture repair yields improved footprint coverage compared with single-row repair. Although it did not reach statistical significance, there was a higher mean yield load in the double-row group., Clinical Relevance: Double-row suture fixation technique for hip abductor tears maximizes strength and footprint coverage of the repair., (Copyright © 2019 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.)

Background: Intestinal parasites are the most common causes of gastrointestinal disease in canine. Stray dogs and wild candies can represent potential reservoirs of enteropathogens to other hosts. Therefore, present study determined the prevalence of intestinal parasites in canine in Mazandaran Province, northern, Iran., Methods: Overall, 58 small intestinal samples of animals (42 stray dogs and 16 jackals) were collected from Oct 2012 to Dec 2013. The intestine contents were studied to detect and identify helminth infections. Then, the helminths were collected and their morphological traits were identified., Results: Overall among infected stray dogs and jackals, 11 species were found. Three species of nematodes, seven species of cestodes and one trematode were observed. The prevalence of gastrointestinal helminths of stray dogs and jackals were 59.5% and 50%, respectively. Among registered zoonotic helminths A. caninum was the predominant parasite both stray dogs and jackals. Interestingly, Spirometra spp. was reported in these animals. Moreover, A. caninum showed a higher percentage rate in center region of province., Conclusion: There are the clear risks of zoonotic helminths parasites infection in this region. Therefore, understanding the epidemiology of zoonotic parasite infection is useful for health care access both domestic animals and humans health., Competing Interests: Conflict interest We have no conflict of interest related to this work.

Background: A more detailed assessment of the anatomy of the entire medial ulnar collateral ligament complex (MUCLC) is desired as the rate of medial elbow reconstruction surgery continues to rise., Purpose: To quantify the anatomy of the MUCLC, including the anterior bundle (AB), posterior bundle (PB), and transverse ligament (TL)., Study Design: Descriptive laboratory study., Methods: Ten unpaired, fresh-frozen cadaveric elbows underwent 3-dimensional (3D) digitization and computed tomography with 3D reconstruction. Ligament footprint areas and geometries, distances to key bony landmarks, and isometry were determined. A surgeon digitized the visual center of each footprint, and this location was compared with the geometric centroid calculated from the outline of the digitized footprint., Results: The mean surface area of the AB was 324.2 mm 2 , with an origin footprint of 32.3 mm 2 and an elongated insertional footprint of 187.6 mm 2 (length, 29.7 mm). The mean area of the PB was 116.6 mm 2 (origin, 25.9 mm 2 ; insertion, 15.8 mm 2 ), and the mean surface area of the TL was 134.5 mm 2 (origin, 21.2 mm 2 ; insertion, 16.7 mm 2 ). The geometric centroids of all footprints could be predicted within 0.8 to 1.3 mm, with the exception of the AB insertion centroid, which was 7.6 mm distal to the perceived center at the apex of the sublime tubercle. While the PB remained relatively isometric from 0° to 90° of flexion ( P = .606), the AB lengthened by 2.2 mm ( P < .001)., Conclusion: Contrary to several historical reports, the insertional footprint of the AB was larger, elongated, and tapered. The TL demonstrated a previously unrecognized expansive soft tissue insertion directly onto the AB, and additional analysis of the biomechanical contribution of this structure is needed., Clinical Relevance: These findings may serve as a foundation for future study of the MUCLC and help refine current surgical reconstruction techniques., Competing Interests: One or more of the authors has declared the following potential conflict of interest or source of funding: This work was funded by internal institutional funds, including the Hospital for Special Surgery Shoulder and Sports Medicine Research Fund and Surgeon in Chief Fund, the Clark Foundation, the Kirby Foundation, and the Gosnell Family. C.L.C. has received financial or material support from Arthrex. J.S.D. is a paid consultant for Arthrex and ConMed Linvatec, is a paid presenter/speaker for Arthrex, receives research support from Arthrex, receives royalties from Biomet, and receives publishing royalties from Wolters Kluwer Health–Lippincott Williams & Wilkins.

α-Lufin, found in Luaf cylindrica seeds, is a type I ribosome inactivating proteins. Cytotoxic effects make it an appropriate candidate for the construction of immunotoxins and conjugates. Because of limited natural resources, recombinant technology is the best approach to achieve large-scale production of plant-based proteins. In the present study, α-lufin protein was expressed in E. coli and the effects of different temperature conditions, SUMO fusion tag, and cultivation strategies on total expression and solubility were investigated. Protein expression was evaluated at different intervals (0, 4, 6, 8, 24 h) postinduction. Our results showed that EnBase had higher eficiency than LB, and maximum solubility and total protein expression were achieved 24 h after induction at 30 °C and 25 °C, respectively. It was shown that SUMO tag is an effective strategy to improve protein solubility.

The steady maturation of computational biomechanics is providing the musculoskeletal health community with exciting avenues for enhancing orthopedic practice and rehabilitation. Computational knee models deliver tools that may improve the efficiency and outcomes of orthopedic research and methods through analysis of virtual surgeries and devices. They also provide insight into the interaction of knee structures and can predict what cannot be directly measured such as loading on our cartilage and ligaments during movement. This project created subject-specific computational knee models of two young adult females using magnetic resonance imaging-derived knee geometries and passive leg motion measured by a motion capture system. The knee models produced passive ligament lengthening patterns similar to experimental measurements available in the literature. The models also predicted cruciate ligament forces during passive flexion with and without applying anterior-posterior tibia forces that were similar to experimental measurements available in the literature. The biomechanics of the posterior oblique ligament (POL) and the anterior cruciate ligament bundles during combined tibia internal-external rotation torque and anterior-posterior forces through deep flexion were then examined. The study showed that the central arm of the POL: (1) produces a maximum constraining force when the knee is at full extension, (2) constrains internal tibial rotation at extension, and (3) constrains posterior tibial translation at extension. The POL reinforces the constraint of the anterior cruciate ligament to internal rotation at extension and provides constraint for posterior tibial translation at extension, a position where the posterior cruciate ligament provides minimal posterior translation constraint., (Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.)

The manipulation of redox potential in secretory pathway by thiol reducing agents can be a strategy to improve the production levels of disulfide-bonded proteins including recombinant antibodies. Here we have studied the influence of cysteamine on viability and the production level of IgG4 in Sp2.0 cells. For this purpose, the recombinant Sp2.0 cells producing an anti CD33 IgG4, were subjected to different concentrations of cysteamine. At concentrations of 2, 4 and 5 mM cysteamine, the secreted levels of IgG4 did not change significantly. However, in concentration of 7 mM cysteamine, a significant decrease was observed in IgG4 levels which may indicate the cytotoxicity of this compound in higher concentrations. Our results show that the cysteamine treatment reduces the cell viability in a dose-dependent manner. Also it was observed that 2 mM cysteamine had no late effect on IgG4 production level and only at day 3, this concentration of cysteamine decreased the cell viability significantly. To test whether the addition of cysteamine can affect the expression level of protein disulfide isomerase, RT-PCR analysis was carried out. The results revealed that cysteamine does not affect the PDI transcription and expression level of IgG4 in this type of recombinant cells.

Radiographic measurements have established a link between meniscus extrusion and meniscus degeneration as well as with knee osteoarthritis. The presented work combines medical imaging with motion capture data from two healthy female subjects to create subject specific knee models that predict tibio-menisco-femoral contact forces and ligament forces during muscle driven simulations of barefoot gait. The developed computational models were used to explore the relationship between the extent of meniscal extrusion and biomechanical function by altering the laxity of the meniscal horn attachments during gait. The extrusion distance increased as laxity increased and the amount of contact force transferred through the menisci during gait decreased rapidly as the meniscal attachments became more lax. Horn attachment lengths that were 20% longer than MRI attachment lengths resulted in an almost complete loss of force transfer through the menisci during the gait cycle. Relatively small changes (2-3mm) in the lengths at which horn bundles first become taut, manifested in large changes in the capacity of the tissue to transmit forces. As meniscal horn attachment laxity increased from 80% to 120% of the MRI measured horn distance, medial meniscus extrusion increased 3.9mm for the first subject and 2.7mm for the second subject. For the same horn laxity changes, the percent of medial tibiofemoral contact force transmitted through the medial meniscus during early stance decreased from 51% to 8% and from 36% to 14% for the two subjects. The results of our study show that increased meniscal extrusion occurs with increased laxity of the meniscal tibia attachments and this increased laxity results in loss of meniscal function., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The strain of Acremonium strictum PTCC 5282 was applied to investigate the biotransformation of androst-1,4-dien-3,17-dione (I; ADD). Microbial products obtained were purified by preparative TLC and the pure metabolites were characterized on the basis of their spectroscopic features (13C NMR, 1H NMR, FTIR, MS) and physical constants (melting points and optical rotations). The 15 alpha-Hydroxyandrost-1,4-dien-3,17-dione (II), 17 beta-hydroxyandrost-1,4-dien-3-one (III), androst-4-en-3,17-dione (IV; AD), 15 alpha-hydroxyandrost-4-en-3,17-dione (V), 15 alpha,17 beta-dihydroxyandrost-1,4-dien-3-one (VI) and testosterone (VII) were produced during this fermentation. Formation of the 15 alpha,17 beta-dihydroxy derivative of ADD is reported for the first time during steroid biotransformation. The bioconversion reactions observed were 1,2-hydrogenation, 15 alpha-hydroxylation and 17-ketone reduction. From the time course profile of this biotransformation, ketone reduction and 1,2-hydrogenation were observed from the first day of fermentation while 15 alpha-hydroxylation occurred from the third day. Optimum concentration of the substrate, which gave the maximum bioconversion efficiency, was 0.5 mg ml(-1) in one batch. The highest yield of the microbial products recorded in this work was achieved within the pH range 6.5-7.3 and at the temperature of 27 degrees C.