The yellow fever (YF) vaccine is usually produced with egg-based methods, which has limitations, including potential adverse effects and low production yields. Alternatively, producing the vaccine using Vero cells or HEK 293 cells can overcome some of these issues, but these methods are significantly more expensive. In the current study, the YF vaccine candidate 17DD virus was produced in primary chicken embryo fibroblast (CEF) cells. The primary CEF cells isolation from eggs was optimized through a two-step process. In the first step, the important parameters that contribute to the development of the egg embryo, such as egg position, relative humidity (RH), and incubation time are optimized. In second step, primary CEF release parameters namely; trypsin volume and incubation temperature are optimized. Both steps were optimized using statistical methods. Further, the seeding cell density of isolated CEF was also optimized. It was observed that 5 x 104 cells/cm2 gave the highest virus titer of 3.89 million PFU/ml. The 17DD yields achieved in primary CEFs are much higher than egg-based production and it is an economically viable method.

The aim of this study was to develop a low-cost and personalized method for external breast prosthesis production.
The projected light method was used for the acquisition of the 3D geometry of the left breast of a healthy 29-year-old woman, 69 kg and 1.69 m. The 3D modeling software Blender was used to make the prosthesis model and mold with adjustments to the model\'s mesh, such as smoothing, assigning thickness, and creating the walls of the prosthesis mold. Two counter-molds were created. The pieces were manufactured on the 3D printer Stella Lite 3 using polylactic acid filament. Finally, the silicone was pigmented, and the mold was filled.
Prototype 1 of the prosthesis was produced using a mold without a counter-mold, which resulted in a prosthesis of 495 g, considered heavy compared to traditional prostheses for the same breast size. To solve this issue, a counter-mold with pins was used to produce prototype 2 with a mass of 393 g, 20.6% lighter than prototype 1. Prototype 3 was made with a central-volume counter-mold and presented a mass of 355 g, a reduction of 28.3% compared to prototype 1. The definitive breast prosthesis was made with the pin counter-mold with a different silicone. It has nipple and areola pigmentation and a mass of 294 g, 25.2% lighter than prototype 2.
The results suggest that the projected light method and additive manufacturing are potential tools for developing external breast prostheses, which may improve the health conditions and quality of life of mastectomized women.

Microwire microelectrode arrays (MEAs) have been a popular low-cost tool for chronic electrophysiological recordings and are an inexpensive means to record the electrical dynamics crucial to brain function. However, both the fabrication and implantation procedures for multi-MEAs on a single rodent are time-consuming and the accuracy and quality are highly manual skill-dependent. To address the fabrication and implantation challenges for microwire MEAs, (1) a computer-aided designed and 3D printed skull cap for the pre-determined implantation locations of each MEA and (2) a benchtop fabrication approach for low-cost custom microwire MEAs were developed. A proof-of-concept design of a 32-channel 4-MEA (8-wire each) recording system was prototyped and tested through Sprague Dawley rat recordings. The skull cap design, based on the CT-scan of a single rat conforms well with multiple Sprague Dawley rats of various sizes, ages, and weight with a minimal bregma alignment error (A/P axis standard error of the mean = 0.25 mm, M/L axis standard error of the mean = 0.07 mm, n = 6). The prototyped 32-channel system was able to record the spiking activities over five months. The developed benchtop fabrication method and the 3D printed skull cap implantation platform would enable neuroscience groups to conduct in-house design, fabrication, and implantation of customizable microwire MEAs at a lower cost than the current commercial options and experience a shorter lead time for the design modifications and iterations.

Reverse transcription-quantitative RT-PCR (RT-qPCR) is a powerful tool for assessing gene transcription levels. The technique is especially useful for measuring estrogen receptor transcript levels as well as gene expression changes in response to estrogen stimulation as it is quick, accurate, and robust and allows the measurement of gene expression in a variety of tissues and cells. This chapter describes the protocols used for RNA extraction and analysis as well as for RT-qPCR assay using hydrolysis (TaqMan-type) probes.

BACKGROUND: Thus far, the hip revision surgery has been widely used and promoted, and the technology has been constantly innovated, such as tissue engineering, 3D printing prosthesis, etc. However, traditional standardized prosthesis, allograft, autograft, bone cement and reinforcing ring are still the main treatment methods in the mainstream pelvic defects classification systems for hip revision. In addition, the mainstream classification systems are still mainly focus on the peri-acetabulum, but less on the large-scale complex pelvic defects that widely affecting the regions far away from the acetabulum, which also have a significant impact on the holistic biomechanical properties of pelvis.
METHODS: After integrating the design experience of custom prostheses and the understanding of biomechanical properties of pelvis, an innovative pelvic defects classification for custom revision was preliminarily proposed, and was practiced in surgeries. Some typical cases were chosen for elucidation in this study, and two observers each evaluated their CT data independently twice. Intraobserver and interobserver agreement were calculated using the kappa statistic to evaluate the reliability. The pelvis defects were classified into five types and two subtypes. The corresponding reconstruction principles, as the main basis to support the classification, were also described in detail. Prosthesis position examination and Harris hip score were utilized to evaluate the clinical outcome.
RESULTS: The installed prostheses resulted in high concordance with preoperative position planning, significantly improved Harris score, low postoperative complication rate and no re-revision case. In addition, The interobserver and intraobserver agreement were both excellent.
CONCLUSIONS: The presenting revision system for complex pelvic defects utilizing 3D-printed custom prosthesis and corresponding classification of pelvic defects can preliminarily guide patients\' grouping and prosthesis design, and may potentially provide an innovative, feasible, and efficient basis for complex total hip arthroplasty (THA) revision.
UNASSIGNED: This study provides a novel method for prosthetic revision of peri-acetabular pelvic defects, and is expected to systematically improve the efficiency of prosthesis design and surgery in clinical practice.

Sperm motility and concentration are commonly evaluated parameters in semen analysis. Those parameters are assessed objectively with commercial instrumentation such as computer-assisted sperm analysis systems (CASA) and hemocytometer. In CASA systems, sperm motility is assessed in the horizontal plane imposed by the stage of the microscope. Thus, there is lack of measurement of the vertical velocity of sperm. The female reproductive tract is a tridimensional space which the sperm traverse to reach the ovum, and there is a need for instruments measuring parameters more relevant to this real-world situation. In this report we describe the design, construction and use of an open-source hardware (OSH) device for evaluation of the vertical velocity of sperm, called UPSPERM. This device was also used to measure sperm concentration, and agreement with hemocytometer was evaluated. Bland-Altman analysis shows good agreement between these two methods of sperm counting. As a first application of UPSPERM, we evaluated the changes in boar sperm motility at distinct pH values between 7.0 and 8.0. The UPSPERM results showed that the vertical velocity of sperm was highest at pH 7.6 and 7.8. We propose that our UPSPERM offers a reliable and affordable option for obtaining measurements of vertical velocity and sperm concentration.

The present study aimed to evaluate the biomechanical behavior of a custom 3D-printed polyetheretherketone (PEEK) condylar prosthesis using finite element analysis and mechanical testing. The Mimics software was used to create a 3D model of the mandible, which was then imported into Geomagic Studio software to perform osteotomy of the lesion area. A customized PEEK condyle prosthesis was then designed and the finite element model of the PEEK condyle prosthesis, mandible and fixation screw was established. The maximum stress of the prosthesis and screws, as well as stress and strain of the cortical and cancellous bones in the intercuspal position, incisal clench, left unilateral molar clench and right unilateral molar clench was analyzed. The biomechanical properties of the prosthesis were studied using two models with different lesion ranges. To simulate the actual clinical situation, a special fixture was designed. The compression performance was tested at 1 mm/min for the condyle prosthesis, prepared by fused deposition modeling (FDM). The results of a finite element analysis suggested that the maximum stress of the condyle was 10.733 MPa and the maximum stress of the screw was 9.7075 MPa; both were far less than the yield strength of the material. The maximum force that the two designed prostheses were able to withstand was 3,814.7±442.6 N (Model A) and 4,245.7±348.3 N (Model B). Overall, the customized PEEK condyle prostheses prepared by FDM exhibited a uniform stress distribution and good mechanical properties, providing a theoretical basis for PEEK as a reconstruction material for repairing the temporomandibular joint.

Orthodontic treatment is more complicated when both soft and hard tissues must be considered because an impacted maxillary canine has important effects on function and esthetics. Compared with extraction of impacted maxillary canines, exposure followed by orthodontic traction can improve esthetics and better protect the patient\'s teeth and alveolar bone. Therefore, in order to achieve desirable tooth movement with minimal unexpected complications, a precise diagnosis is indispensable to establish an effective and efficient force system. In this report, we describe the case of a 31-year-old patient who had a labio-palatal horizontally impacted maxillary left canine with a severe occlusal alveolar bone defect and a missing maxillary left first premolar. Herein, with the aid of three-dimensional imaging, sequential traction was performed with a three-directional force device that finally achieved acceptable occlusion by bringing the horizontally impacted maxillary left canine into alignment. The maxillary left canine had normal gingival contours and was surrounded by a substantial amount of regenerated alveolar bone. The 1-year follow-up stability assessment demonstrated that the esthetic and functional outcomes were successful.

To use computational simulations to compare the hemodynamic characteristics of a classic bifurcated stent-graft to an equally long endograft design with \"dog bone\"-shaped limbs (DB), which have large diameter proximal and distal ends and significant narrowing at the midsection to accommodate aneurysms with an extremely narrow bifurcation.
A 3-dimensional model was constructed using commercially available validated software. Inlet and outlet diameters were 28 and 14 mm, respectively. The total length of both models was kept constant to 180 mm, but the main body of the DB model was 20 mm shorter than the bifurcated endograft. The iliac limbs of the DB model had a 9-mm stenosis over a 30-mm segmental length in the midsection. Flow was quantified by time-averaged wall shear stress, oscillatory shear index (OSI), and relative residence time (RRT). The displacement forces in newtons (N) and maximum wall shear stress (WSS) in pascals (Pa) were compared during a cardiac cycle at 3 segments (main body, bifurcation, and iliac limbs) of both models with computational fluid dynamics analysis.
The DB accommodation was associated with higher forces at the main body (range 3.15-4.9 Ν) compared with the classic configuration (1.56-2.34 N). On the contrary, the forces at the bifurcation (3.81-5.98 vs 3.76-5.54 N) and at the iliac limbs (0.34-0.85 vs 0.49-0.74 N) were comparable for both models. Accordingly, maximum WSS was detected at the iliac sites for both models throughout the cardiac cycle. The highest values were detected at peak systole and equaled 26.6 and 12 Pa for the DB and bifurcated configurations, respectively. The narrow segments in the DB model displayed high stress values but low OSI and very low RRT.
The DB accommodation seems to correlate with higher displacement forces at the main body and higher stresses at the iliac limbs. Consequently, regular imaging follow-up of the DB design deems necessary to delineate its mid- and long-term clinical performance.

The evaluation concerns on custom design of implant for bony defect focus on engineered transformation of clinical needs, pre-operation performance verification and clinical validation. These concerns refer to manipulation on imageology information, tissue adhesion and fixation, surgery planning and instruments, design specification and instruction, finite element analysis, bench testing, around market clinical research et al. The evaluation integrates adaptive risk evaluation methods such as biomechanics, mechanobiology, blind crossover reproducility and dynamic design of clinical study et al.