Cutting rodent's bone ends and irrigation of the medullary canal is the common method used for cells collection in allogenic transplantation, however it does not yield sufficient cells for autologous transplantation. The aim of this experiment was to establish and validate a method for bone marrow collection for autologous MSCs transplantation. Two collection methods were examined: 1) Transection of the bone ends and irrigation of the medullary canal, 2) Trephining of the bone with a hypodermic needle without aspiration. Then cell harvesting was compared in the idealised laboratory situation and under simulated surgery. First, two lower limbs were harvested from the same rat cadaver for comparison, bone marrow in one limb was collected by cutting the femoral head and the distal tibia and irrigation of the canal through drilled holes at the distal end of the femur and proximal end of the tibia. Other limb, hypodermic needle was used as a trephining tool into the medullary canal multiple times without applying negative pressure and rinsed from inside and outside. Second, bone marrow was harvested from another rat's cadaver in the surgery room to simulate the conditions needed for autologous transplantation. The number of cells from irrigation method was 1.28*106 cells, whereas that from trephining method reached 17*106. The number cells from the bone marrow harvested in the surgery room was found 29.6*106. We report a novel technique for harvesting cells for autologous cell therapy from only one limb. A significantly larger number of cells from bone marrow could be collected using the needle trephining method. There is no negative effect on the viability of cells after bone marrow harvesting in the surgery room

To quantify bone-nail fit in response to varying nail placements by entry point translation in straight antegrade humeral nailing using three-dimensional (3D) computational analysis. CT scans of ten cadaveric humeri were processed in 3D Slicer to obtain 3D models of the cortical and cancellous bone. The bone was divided into individual slices each consisting of 2% humeral length (L) with the centroid of each slice determined. To represent straight antegrade humeral nail, a rod consisting of two cylinders with diameters of 9.5mm and 8.5mm and length of 0.22L mm and 0.44L mm respectively joined at one end was modelled. The humeral head apex (surgical entry point) was translated by 1mm in both anterior-posterior and medio-lateral directions to generate eight entry points. Total nail protrusion surface area, maximum nail protrusion distance into cortical shell and top, middle, bottom deviation between nail and intramedullary cavity centre were investigated. Statistical analysis between the apex and translated entry points was conducted using paired t-test. A posterior-lateral translation was considered as the optimal entry point with minimum protrusion in comparison to the anterior-medial translation experiencing twice the level of protrusion. Statistically significant differences in cortical protrusion were found in anterior-medial and posterior-lateral directions producing increased and decreased level of protrusion respectively compared to the apex. The bottom anterior-posterior deviation distance appeared to be a key predictor of cortical breach with the distal nail being more susceptible. Furthermore, nails with anterior translation generated higher anterior-posterior deviation (>4mm) compared to posterior translation (<3mm). Aside from slight posterolateral translation of the entry point from the apex, inclusion of a distal posterior-lateral bend into current straight nail design could improve nail fitting within the curved humeral bone, potentially improving distal working length within the flat and narrow medullary canal of the distal humeral shaft

In 2019, Lin et al. published a proof-of-concept study of electrical impedance spectroscopy as a simple and low-cost method to characterize progression of fracture repair (Lin et al., Sci Rep 2019). However, the electrical impedance sensors were placed in the fracture site which may impair the transfer to clinical use. To further explore the concept of monitoring fracture healing by electrical impedance spectroscopy, we established a tibial fracture model in the rabbit where sensors are positioned in proximity to the fracture site but without being placed in the fracture site. The aim of this pilot study was to explore whether distinct patterns of electrical impedance would evolve as tibial fractures in rabbits were evaluated until radiographic signs of healing. Approval was granted from the Inspectorate of the Animal Experimentation under the Danish Ministry of Justice. Four rabbits were anaesthetized, and in each rabbit a tibial osteotomy was made and stabilized by an external fixator. Electrical impedance was measured immediately postoperative and hereafter daily until euthanization after 3 weeks. Recordings were obtained within a wide frequency range (10 Hz to 1 MHz) from an inner electrode placed into the medullary canal and an outer electrode placed extracortical on the lateral with a distance of 3 mm to the defect. A similar pattern of electrical impedance over time was observed in the four rabbits. During the very early stages of fracture healing, an initial fluctuation in electrical impedance occurred. However, after 10 days the curves revealed a steady daily increase in electrical impedance. The first radiological signs of bone healing were detected after 14 days and progressed in all four rabbits in accordance with increments in the electrical impedance until termination of the pilot study after 21 days. Consistent electrical impedance patterns were detected during bone healing in a pilot study of four rabbits. Further research is needed to explore whether the presented method of electrical impedance measurements can be used to monitor bone healing over time

Electrical impedance spectroscopy measurements might be used for real-time monitoring of bone healing. Differences in electrical characteristics of different tissues during fracture healing can provide information of the tissue composition within the fracture region. This concept embraces the digital revolution of orthopaedics allowing for a sensor-based and home-based monitoring of bone healing. Furthermore, real-time monitoring will allow individualized and timely treatment adjustments to prevent bone healing complications. However, electrodes must be placed at a distance to the fracture site in order not to interfere with bone healing. Therefore, we investigated whether longitudinal and transverse electrical impedance measurements from electrodes placed at a distance to the bone defect can detect differences between intact bone and bone defects in vivo. Approval was granted from the Inspectorate of the Animal Experimentation under the Danish Ministry of Justice. Six rabbits were anaesthetized and had both tibias subjected to an osteotomy protocol where first the medial, then the lateral, and final the posterior cortex were removed resulting in a complete 2 mm bone defect. Electrical impedance was measured prior to and after each step of the osteotomy protocol. Recordings were obtained at different frequencies (10 Hz to 1 MHz) from an inner electrode placed into the medullary canal and two different electrodes placed extracortical on the lateral and posterior bone with a distance of 5 mm to the defect. For each rabbit, one tibia had measurements with a free inner electrode and the other tibia had measurements both with a nail and an isolated nail. For all tibias, the intact bone resulted in higher impedance compared with the complete defect, and this difference was most pronounced in the frequency range of 1 kHz to 100 kHz. This applied for all types of internal electrodes including electrode, nail, isolated nail. The isolated nail showed the biggest impedance difference between the intact bone and the complete defect. Incomplete bone defects had lower impedance compared with intact bone, but no consistent pattern for differences in impedance was observed between the different applied defects. Consistent impedance differences between intact bone and complete defects were detected in-vivo in rabbits. Further research is needed to explore whether the presented method of electrical impedance measurements can be used to characterize bone healing over time

Summary Statement. This 3-dimensional CT study on cadaveric proximal ulna provides further insight into the size and geometry of the proximal ulna intramedullary cavity with potential applications to design and sizing of proximal ulna components. Introduction. Total elbow arthroplasty (TEA) is an established treatment for varying pathologies of the elbow with very good functional outcomes. Optimal fit of ulna components in TEA is predicated on a detailed appreciation of the 3-dimensional anatomy of the proximal ulna intra-medullary cavity, but literature remains scarce. Three-dimensional (3D) models of the proximal ulna have been constructed using computed tomography (CT) programs, accurately defining the angular relations with the cross-sectional extra-medullary dimensions. However, current CT-based thresholding techniques lack accuracy in differentiating cortical from cancellous bone in the metaphyseal region, and thus cannot properly define the intra-medullary region of uncored proximal ulnae. We investigate the geometric dimensions of the proximal ulna intra-medullary cavity using CT studies of cored cadaveric ulnae. Patients & Methods. Three-dimensional models based on CT data of 17 manually cored cadaveric proximal ulnae were created using Matlab program and analyzed. The cadaveric specimens were dissected to identify the medullary canal and coring of the canal of the proximal ulna to exclude the cancellous bone was performed as for ulna canal preparation during TEA. Using the center of a circle fitted over the trochlear ridge as the origin, the diameter, coronal and sagittal angulation of the canal were determined. The diameter, posterior and lateral offsets of the proximal medullary canal were plotted against the normalised axial distance from the origin which was represented in terms of R, the radius of the circle fitted to the trochlear ridge. The normalization of the axial distance was done to compensate for the varying individual ulna lengths. Results. The mean of the radius of the circle which was fitted to the trochlear ridge, R, was found to be 16.64 ± 2.71 mm. The minimum diameter of the canal increased from the origin and peak at 2.3R (just distal to the coronoid process) with a value of 7.08 ± 1.74 mm, decreasing in a linear fashion to 4.48 ± 0.91 mm at 6R. There was an increasing posterior offset which was linear in nature (R. 2. = 0.954) up to 6R (approximately 97mm from the origin). Likewise, the lateral offset increased in a linear fashion (R. 2. = 0.996). Discussion. The findings of our study most likely represent the effective minimum diameter for optimal implant placement, which are consistent with previously documented cadaveric dimensions. The findings also support the current tapered component design of the proximal ulna stemmed implants without the need for significant angulation. Conclusion. Our study provides further insight into the size and geometry of the proximal ulna intramedullary cavity with potential applications to design and sizing of proximal ulna components

Background. The treatment of olecranon fractures frequently involves the use of tension band fixation. Although associated with high union rates, this method has a high incidence of morbidity associated with soft tissue compromise and limitation of range of movement requiring frequent re-operation for removal of metal. Objectives. We describe the use of a simple jig to ensure intramedullary placement of longitudinal K-wires and compare the accuracy of placement of wires using this device with the traditional free hand method. Methods & Materials. 10 orthopaedic surgical trainees passed 2 longitudinal k wires into synthetic ulnae. This was done using the free hand method and then plastic jig. The ulnae were then sectioned to identify the position of the wires relative to the intramedullary canal. Results. Of the free hand wires, only 11 of 20 were found to be intramedullary versus 20 of 20 in the group passed using the jig. (p<0.001). The mean distance from the center of the ulnar canal was 6.5 mm in the freehand group (range 1-18mm, SD 6.1mm) and 1.6 mm in the O-Jig group (range 1-2mm, SD 0.5mm). The difference in the mean distance from the centre was 4.9mm which was significant (p< 0.001). Conclusions. We found the distance from the centre of the medullary canal, the range and standard deviations of wire positions to be significantly more precise when the jig was used

Summary Statement. We used three-dimensional software to assess different anatomic variables in the femur. The canal of Femur twisted slightly below the lesser trochanter in cases with a larger angle of anteversion. Introduction. Accurate positioning of the joint prosthesis is essential for successful total hip arthroplasty (THA). To aid in tailoring of the prosthesis, we used three-dimensional software to assess different anatomic variables in the femur. Patients & Methods. We used CT imaging data of the unaffected normal side of the 25 patients (22 females, age range 30 to 81 years) who underwent THA in 2012 in our hospital. The femur was reconstructed from CT data and measured using three-dimensional modeling software (Mimics 16.0 Materialise, Leuven, Belgium). We measured ellipse fitting to the medullary canal in the axial plane of the femur at 20-mm intervals. The angle between the major axis of those ellipses and the axis of the femoral neck was measured and expressed as the canal rotation. The distance between the lesser trochanter and the center of the femoral head was measured along the Z axis. Results. The major axes of the ellipses direct to medial, front and medial side in the level of epiphysis, above isthmus and distal portion respectively in all cases. The maximum rotated level was above isthmus. The rotation angle in the proximal portion ranged from 36 to 84 degrees (mean, 60.6 degrees, SD ± 12.1). The rotation angle of the distal portion ranged from 71 to 95 degrees (mean, 86.1 degrees, SD ± 6.1). Discussion/Conclusion. The torsion of the canal varied more widely between individuals in the proximal portion than did the distal portion. In addition, the torsion of the proximal aspect, although more variable, was on average smaller when the angle of anteversion was large. Because the canal twisted slightly below the lesser trochanter in cases with a larger angle of anteversion, it is suggested that attention to the degree of anteversion of a flat prosthesis stem is warranted

This study investigated the quality and quantity of healing of a bone defect following intramedullary reaming undertaken by two fundamentally different systems; conventional, using non-irrigated, multiple passes; or suction/irrigation, using one pass. The result of a measured re-implantation of the product of reaming was examined in one additional group. We used 24 Swiss mountain sheep with a mean tibial medullary canal diameter between 8 mm and 9 mm. An 8 mm ‘napkin ring’ defect was created at the mid-diaphysis. The wound was either surgically closed or occluded. The medullary cavity was then reamed to 11 mm. The Reamer/Irrigator/Aspirator (RIA) System was used for the reaming procedure in groups A (RIA and autofilling) and B (RIA, collected reamings filled up), whereas reaming in group C (Synream and autofilling) was performed with the Synream System. The defect was allowed to auto-fill with reamings in groups A and C, but in group B, the defect was surgically filled with collected reamings. The tibia was then stabilised with a solid locking Unreamed Humerus Nail (UHN), 9.5 mm in diameter. The animals were killed after six weeks. After the implants were removed, measurements were taken to assess the stiffness, strength and callus formation at the site of the defect. There was no significant difference between healing after conventional reaming or suction/irrigation reaming. A significant improvement in the quality of the callus was demonstrated by surgically placing captured reamings into the defect using a graft harvesting system attached to the aspirator device. This was confirmed by biomechanical testing of stiffness and strength. This study suggests it could be beneficial to fill cortical defects with reaming particles in clinical practice, if feasible

Introduction. Hip resurfacing arthroplasty (HRA) with metal-on-metal bearing is currently emerging as a major evolution of hip arthroplasty. It could be an alternative to total hip arthroplasty. HRA also may allow young patients to perform high sports activities. It preserves proximal femoral bone stock and keeps the medullary canal intact. A large diameter of the femoral head provides wider range of the hip motion. Incidence of dislocation is very low. Most of HRA have been reported for young patients with stage of osteoarthritis. But, reports of HRA for osteonecrosis of the femoral head (ONFH) are rare and the outcomes are uncertain. Methods. This study was performed to introduce our indication and technique of HRA for the patients with ONFH and to assess the outcomes after a minimum 5-year follow up. After the biomechanical study, we set our own indications for HRA as follows: when the size of a lesion was less than 50% of the entire head and the rim at the head-neck junction was intact at least to a 5 mm above from the head-neck junction. From December 1998 to July 2004; 157 hips (139 patients) underwent HRA under the diagnosis of ONFH. Their mean age was 37 years. One hundred and fourteen hips were in men and 25 hips were in women. The average follow-up period was 88 months (range, 60 to 128 months). Clinical outcomes were evaluated with Harris Hip Scores, hip or thigh pain, limb length discrepancy, and range of motion. As a radiological evaluation, the preoperative size and staging of the lesion, patterns of bone remodeling, and complications such as femoral neck fracture, neck narrowing, loosening, and osteolysis were analyzed. Results. Sixty-two percent of the hips were staged as Steinberg IV or more. The size of the necrotic lesion in magnetic resonance imaging ranged from 23 to 50%. The Harris Hip Score increased from 77 points pre-operatively to 98 points at the final visit. All range of motions significantly improved after the operation. No patient complained of limb length discrepancy. Twenty-eight hips showed painless radiographic changes including impingement (22 hips), focal radiolucency (4 hips), and neck narrowing (2 hips). All patients showed no clinical symptoms. There was no detectable wear, dislocation, femoral neck fracture, or other complications. No revision or additional surgery was required in all patients. Conclusion. Even for the patients with ONFH, the overall minimum 5-year outcomes of HRA are promising and superior to other reports of conventional THA. Patient satisfaction is extremely high. A strict application of indications and meticulous surgical technique are critical for these successful results

Aims

This study intended to investigate the effect of vericiguat (VIT) on titanium rod osseointegration in aged rats with iron overload, and also explore the role of VIT in osteoblast and osteoclast differentiation.

The feasibility of bone transport with bone substitute and the factors which are essential for a successful bone transport are unknown. We studied six groups of 12 Japanese white rabbits. Groups A to D received cylindrical autologous bone segments and groups E and F hydroxyapatite prostheses. The periosteum was preserved in group A so that its segments had a blood supply, cells, proteins and scaffold. Group B had no blood supply. Group C had proteins and scaffold and group D had only scaffold. Group E received hydroxyapatite loaded with recombinant human bone morphogenetic protein-2 and group F had hydroxyapatite alone.

Distraction osteogenesis occurred in groups A to C and E which had osteo-conductive transport segments loaded with osteo-inductive proteins. We conclude that scaffold and proteins are essential for successful bone transport, and that bone substitute can be used to regenerate bone.

Objectives

We investigated the effects on fracture healing of two up-regulators of inducible nitric oxide synthase (iNOS) in a rat model of an open femoral osteotomy: tadalafil, a phosphodiesterase inhibitor, and the recently reported nutraceutical, COMB-4 (consisting of L-citrulline, Paullinia cupana, ginger and muira puama), given orally for either 14 or 42 days.

Hydroxyapatite-coated standard anatomical and customised femoral stems are designed to transmit load to the metaphyseal part of the proximal femur in order to avoid stress shielding and to reduce resorption of bone. In a randomised in vitro study, we compared the changes in the pattern of cortical strain after the insertion of hydroxyapatite-coated standard anatomical and customised stems in 12 pairs of human cadaver femora. A hip simulator reproduced the physiological loads on the proximal femur in single-leg stance and stair-climbing. The cortical strains were measured before and after the insertion of the stems.

Significantly higher strain shielding was seen in Gruen zones 7, 6, 5, 3 and 2 after the insertion of the anatomical stem compared with the customised stem. For the anatomical stem, the hoop strains on the femur also indicated that the load was transferred to the cortical bone at the lower metaphyseal or upper diaphyseal part of the proximal femur.

The customised stem induced a strain pattern more similar to that of the intact femur than the standard, anatomical stem.

In a rabbit model we investigated the efficacy of a silk fibroin/hydroxyapatite (SF/HA) composite on the repair of a segmental bone defect. Four types of porous SF/HA composites (SF/HA-1, SF/HA-2, SF/HA-3, SF/HA-4) with different material ratios, pore sizes, porosity and additives were implanted subcutaneously into Sprague-Dawley rats to observe biodegradation. SF/HA-3, which had characteristics more suitable for a bone substitite based on strength and resorption was selected as a scaffold and co-cultured with rabbit bone-marrow stromal cells (BMSCs). A segmental bone defect was created in the rabbit radius. The animals were randomised into group 1 (SF/HA-3 combined with BMSCs implanted into the bone defect), group 2 (SF/HA implanted alone) and group 3 (nothing implanted). They were killed at four, eight and 12 weeks for visual, radiological and histological study.

The bone defects had complete union for group 1 and partial union in group 2, 12 weeks after operation. There was no formation of new bone in group 3. We conclude that SF/HA-3 combined with BMSCs supports bone healing and offers potential as a bone-graft substitute.

Allograft bone is widely used in orthopaedic surgery, but peri-operative infection of the graft remains a common and disastrous complication. The efficacy of systemic prophylactic antibiotics is unproven, and since the graft is avascular it is likely that levels of antibiotic in the graft are low.

Using an electrical potential to accelerate diffusion of antibiotics into allograft bone, high levels were achieved in specimens of both sheep and human allograft. In human bone these ranged from 187.1 mg/kg in endosteal (sd 15.7) to 124.6 (sd 46.2) in periosteal bone for gentamicin and 31.9 (sd 8.9) in endosteal and 2.9 (sd 1.1) in periosteal bone for flucloxacillin. The antibiotics remained active against bacteria in vitro after iontophoresis and continued to elute from the allograft for up to two weeks.

Structural allograft can be supplemented directly with antibiotics using iontophoresis. The technique is simple and inexpensive and offers a potential means of reducing the rate of peri-operative infection in allograft surgery. Iontophoresis into allograft bone may also be applicable to other therapeutic compounds.

We investigated the effect of pre-heating a femoral component on the porosity and strength of bone cement, with or without vacuum mixing used for total hip replacement.

Cement mantles were moulded in a manner simulating clinical practice for cemented hip replacement. During polymerisation, the temperature was monitored. Specimens of cement extracted from the mantles underwent bending or fatigue tests, and were examined for porosity.

Pre-heating the stem alone significantly increased the mean temperature values measured within the mantle (+14.2°C) (p < 0.001) and reduced the mean curing time (−1.5 min) (p < 0.001). The addition of vacuum mixing modulated the mean rise in the temperature of polymerisation to 11°C and reduced the mean duration of the process by one minute and 50 seconds (p = 0.01 and p < 0.001, respectively). In all cases, the maximum temperature values measured in the mould simulating the femur were < 50°C. The mixing technique and pre-heating the stem slightly increased the static mechanical strength of bone cement. However, the fatigue life of the cement was improved by both vacuum mixing and pre-heating the stem, but was most marked (+ 280°C) when these methods were combined.

Pre-heating the stem appears to be an effective way of improving the quality of the cement mantle, which might enhance the long-term performance of bone cement, especially when combined with vacuum mixing.

Interfacial defects between the cement mantle and a hip implant may arise from constrained shrinkage of the cement or from air introduced during insertion of the stem. Shrinkage-induced interfacial porosity consists of small pores randomly located around the stem, whereas introduced interfacial gaps are large, individual and less uniformly distributed areas of stem-cement separation. Using a validated CT-based technique, we investigated the extent, morphology and distribution of interfacial gaps for two types of stem, the Charnley-Kerboul and the Lubinus SPII, and for two techniques of implantation, line-to-line and undersized.

The interfacial gaps were variable and involved a mean of 6.43% (sd 8.99) of the surface of the stem. Neither the type of implant nor the technique of implantation had a significant effect on the regions of the gaps, which occurred more often over the flat areas of the implant than along the corners of the stems, and were more common proximally than distally for Charnley-Kerboul stems cemented line-to-line. Interfacial defects could have a major effect on the stability and survival of the implant.

A complete cement mantle is important for the longevity of a total hip replacement. In the minimally-invasive direct anterior approach used at the Innsbruck University hospital, the femoral component has to be inserted into the femoral canal by an angulated movement. In a cadaver study, the quality and the extent of the cement mantle surrounding 13 Exeter femoral components implanted straight through a standard anterolateral transgluteal approach were compared with those of 13 similar femoral components implanted in an angulated fashion through a direct anterior approach. A third-generation cementing technique was used. The inner and outer contours of the cement mantles was traced from CT scans and the thickness and cross-sectional area determined.

In no case was the cement mantle incomplete. The total mean thickness of the cement mantle was 3.62 mm (95% confidence interval 3.59 to 3.65). The mean thickness in the group using the minimally-invasive approach was 0.16 mm less than that in the anterolateral group. The distribution of the thickness was similar in the two groups. The mean thickness was less on the anteromedial and anterolateral aspect than on the posterior aspect of the femur.

There is no evidence that the angulated introduction of Exeter femoral components in the direct anterior approach in cadavers compromises the quality, extent or thickness of the cement mantle.

The use of impaction bone grafting during revision arthroplasty of the hip in the presence of cortical defects has a high risk of post-operative fracture. Our laboratory study addressed the effect of extramedullary augmentation and length of femoral stem on the initial stability of the prosthesis and the risk of fracture.

Cortical defects in plastic femora were repaired using either surgical mesh without extramedullary augmentation, mesh with a strut graft or mesh with a plate. After bone impaction, standard or long-stem Exeter prostheses were inserted, which were tested by cyclical loading while measuring defect strain and migration of the stem.

Compared with standard stems without extramedullary augmentation, defect strains were 31% lower with longer stems, 43% lower with a plate and 50% lower with a strut graft. Combining extramedullary augmentation with a long stem showed little additional benefit (p = 0.67). The type of repair did not affect the initial stability. Our results support the use of impaction bone grafting and extramedullary augmentation of diaphyseal defects after mesh containment.

The treatment of chronic osteomyelitis often includes surgical debridement and filling the resultant void with antibiotic-loaded polymethylmethacrylate cement, bone grafts or bone substitutes. Recently, the use of bioactive glass to treat bone defects in infections has been reported in a limited series of patients. However, no direct comparison between this biomaterial and antibiotic-loaded bone substitute has been performed.

In this retrospective study, we compared the safety and efficacy of surgical debridement and local application of the bioactive glass S53P4 in a series of 27 patients affected by chronic osteomyelitis of the long bones (Group A) with two other series, treated respectively with an antibiotic-loaded hydroxyapatite and calcium sulphate compound (Group B; n = 27) or a mixture of tricalcium phosphate and an antibiotic-loaded demineralised bone matrix (Group C; n = 22). Systemic antibiotics were also used in all groups.

After comparable periods of follow-up, the control of infection was similar in the three groups. In particular, 25 out of 27 (92.6%) patients of Group A, 24 out of 27 (88.9%) in Group B and 19 out of 22 (86.3%) in Group C showed no infection recurrence at means of 21.8 (12 to 36), 22.1 (12 to 36) and 21.5 (12 to 36) months follow-up, respectively, while Group A showed a reduced wound complication rate.

Our results show that patients treated with a bioactive glass without local antibiotics achieved similar eradication of infection and less drainage than those treated with two different antibiotic-loaded calcium-based bone substitutes.

Cite this article: Bone Joint J 2014; 96-B:845–50.