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Distal biceps tendon ruptures have been reported to be associated with narrowing of the proximal radioulnar space. There have been no studies that three-dimensionally measured the distance between the bicipital tuberosity and the proximal ulna during active motion. The purpose of this study was to three-dimensionally measure the proximal radioulnar space during active forearm pronation in healthy subjects. Five healthy volunteers (10 forearms) were recruited for this study. They consisted of all males with a mean age of 37 years (range, 34-46 years). Lateral fluoroscopy of forearm rotation from maximum supination to maximum pronation was recorded for both forearms. Three-dimensional forearm kinematics were determined using model-image registration techniques with fluoroscopic images and CT-derived bone models, and the closest distance between the bicipital tuberosity and the proximal ulna was computed at each 30° increment of radial axial rotation relative to the distal humerus. The distance between the bicipital tuberosity and the proximal ulna decreased with pronation, reaching a minimum value at 90° of radial rotation (average 4.6 ± 1.3 mm), then increased with further rotation to maximum pronation (P = 0.004). The clearance between the proximal radioulnar space and the distal biceps tendon is very small (<1mm). Hypertrophy of the bicipital tuberosity or tendon can induce impingement and lead to tendon rupture., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Microtensile bond strength (μTBS) test was introduced in 1994. Since then, it has been utilized profoundly across many bond strength testing laboratories, making it currently one of the most standard and versatile bond strength test. Although it is a static and strength-based method, together with the morphological and spectroscopic investigations, it has been contributing immensely in the advancement of dentin adhesive systems. μTBS test has a greater discriminative capability than the traditional macro-shear bond test. During the early stage of its development, the authors predicted that this testing method would enable evaluation of the adhesive performances of resins to excavated carious or sclerotic dentin and the regional bond strengths of various portions of the cavity. In addition, they also stated the possibility of comparing the long-term stability of resin adhesion at various portions of the cavity walls on teeth extracted at various times after insertion of bonded restorations. In this review, we discussed the historical background, inception and the application of the μTBS test and proposed directions for further improvement of this testing method., (© 2019 Published by Elsevier Ltd on behalf of The Japanese Association for Dental Science This is an.)

This study aimed to evaluate the bonding performance of a new one-step self-etching adhesive system containing a novel hydrophilic amide monomer. Clearfil Universal Bond Quick (CUB) and Clearfil Megabond 2 (CMB) were used as the one-step and two-step adhesive systems, respectively. Flat dentin surfaces of human premolars were exposed using #600 SiC (silicon carbide) and bonded with the respective adhesives of each system. The teeth were sectioned to obtain beams (1 mm × 1 mm) after 24 h of water storage. The mean bond strength and standard deviations (MPa) on an occlusal surface were as follows: CUB: 45.9 ± 19.7 and CMB: 67.9 ± 25.3. The values for cervical ones were CUB: 56.0 ± 20.3 and CMB: 67.6 ± 16.0, respectively. In both conditions, the microtensile bond strength (μTBS) value was lower than that of CMB. As seen during the microscopic observation, no adhesive failure was observed after μTBS testing because CUB formed a firm and tight adhesive interface.

Background: Type 2 failure is a big issue after suture-bridge rotator cuff repair, which may be because of stress concentration at the medial row stitches. We have been performing medial knot-tying after suture-bridge lateral row repair to avoid the stress concentration. This study aimed to evaluate clinical and radiological outcomes after arthroscopic rotator cuff repair using this technique., Hypothesis: This technique would yield better radiological outcomes with a reduced type 2 failure rate compared with reported outcomes after conventional suture-bridge repair., Study Design: Case series; Level of evidence, 4., Methods: The inclusion criteria of this study were (1) full-thickness tears, (2) primary surgery, and (3) minimum 2-year follow-up with pre- and postoperative magnetic resonance imaging (MRI). We investigated active ranges of motion (forward elevation and external rotation), as well as the Japanese Orthopaedic Association (JOA) and University of California, Los Angeles (UCLA), scores preoperatively and at the final follow-up., Results: This study included 384 shoulders in 373 patients (205 men and 168 women) with a mean age of 65 years (range, 24-89 years) at the time of surgery. The mean follow-up was 29 months (range, 24-60 months). There were 91 small, 137 medium, 121 large, and 35 massive tears. Postoperative MRI scans demonstrated successful repair in 324 shoulders (84.4%, group S) and retear in 60 shoulders (15.6%). Among 60 retears, 40 shoulders (67%) had type 1 failure (group F1) and 20 shoulders (33%) had type 2 failure (group F2). Forward elevation and external rotation significantly improved after surgery ( P < .001 for both). Postoperative JOA and UCLA scores in group F2 were significantly lower than those in the other groups., Conclusion: The medial knot-tying after suture-bridge lateral row repair demonstrated excellent functional and radiological outcomes after surgery, with a retear rate of 15.6%. The type 2 failure showed significantly inferior functional outcomes; however, the rate of type 2 failure was less relative to previous studies using conventional suture-bridge techniques. Our technique could be a good alternative to conventional suture-bridging rotator cuff repair because it may reduce the rate of postoperative type 2 failure.

Background: Superior labral anterior-posterior (SLAP) lesions are common among elite gymnasts and throwing athletes. Although SLAP lesions in throwers are well-described in the literature, no study has described the characteristics of SLAP lesions in gymnasts. We aimed to reveal the characteristics of SLAP lesions in gymnasts by comparing the location and extension of these lesions between gymnasts and throwers., Hypothesis: The location and arc of SLAP lesions in gymnasts will be different from those in throwing athletes., Study Design: Case series; Level of evidence, 4., Methods: This study included 27 shoulders in 20 males and 3 females with a mean ± SD age of 20 ± 2.5 years (range, 16-25 years). We performed debridement alone for shoulders with a stable lesion. Anterior and/or posterior labral repair was added for unstable SLAP lesions depending on the extension and stability of the lesions. We investigated symptoms, onset, return to sport (based on patient records), and subjective shoulder values. SLAP lesions were evaluated through use of the Snyder classification. The location and arc of SLAP lesions were determined from surgical records and videos and described by use of the right shoulder clockface method. During the same period, 65 baseball players (65 shoulders; all males; mean age, 23 ± 7.0 years; range, 16-44 years) underwent arthroscopic SLAP surgery. We compared the location and arc of SLAP lesions between gymnasts and baseball players., Results: Symptoms during gymnastics included pain (100%), apprehension (48%), or catching (11%). We found that 20 shoulders had symptom onset during gymnastics, most commonly during rings events. Type II SLAP lesions were found in 17 shoulders, type III in 2 shoulders, and type IV in 8 shoulders. The mean center of SLAP lesions was at the 11:40 clockface position in 27 gymnasts and 10:40 clockface position in 65 baseball players, and the difference was statistically significant ( P < .001). The mean arc of SLAP lesions was 125° in gymnasts and 140° in baseball players, and the difference was not significant. We performed debridement in 2 shoulders (7%) and labral repair in 25 shoulders (93%). After surgery, all patients returned to gymnastics. The mean subjective shoulder value was 35 (range, 10-90) preoperatively and 76 (range, 40-100) postoperatively., Conclusion: SLAP lesions in gymnasts were significantly located anteriorly compared with those in baseball players. All patients returned to gymnastics after arthroscopic surgery. Secure repair of SLAP lesions may be important for good surgical outcomes, because 50% of patients experienced preoperative shoulder apprehension., Competing Interests: The authors declared that there are no conflicts of interest in the authorship and publication of this contribution. AOSSM checks author disclosures against the Open Payments Database (OPD). AOSSM has not conducted an independent investigation on the OPD and disclaims any liability or responsibility relating thereto., (© The Author(s) 2020.)

Background: No reports have been published on the clinical outcomes, including return to sport, after rotator cuff repair in middle-aged and elderly swimmers with rotator cuff tears., Purpose: To retrospectively investigate clinical outcomes and return to sport after arthroscopic rotator cuff repair in middle-aged and elderly swimmers., Study Design: Case series; Level of evidence, 4., Methods: Patients who underwent arthroscopic rotator cuff repair and met the following criteria were included: (1) age older than 45 years, (2) rotator cuff tears confirmed during surgery, (3) arthroscopic suture bridge rotator cuff repair, (4) primary surgery, and (5) swam more than once a week before surgery. The exclusion criteria were as follows: (1) irreparable large and massive tears, (2) shoulder instability, (3) arthritis or rheumatic disorders, or (4) less than 2-year follow-up. We investigated functional outcomes including range of motion; the University of California, Los Angeles (UCLA) score; the American Shoulder and Elbow Surgeons (ASES) score; return to swimming; and the return rates for each style of swimming. In addition, the functional outcomes and findings of magnetic resonance imaging were compared between the group with complete return and the group with incomplete or failed return., Results: A total of 32 shoulders in 31 middle-aged and elderly swimmers (5 males, 26 females) were included. The mean age was 65 years (range, 47-78 years), and the mean follow-up was 47 months (range, 24-86 months). Return to swimming was achieved in 31 (97%) shoulders at a mean of 8 months (range, 3-24 months), and complete return was achieved in 18 (56%) shoulders at a mean of 12 months (range, 3-24 months). The return rate was 97% for freestyle, 83% for breaststroke, 74% for backstroke, and 44% for butterfly stroke. Postoperative UCLA and ASES scores were significantly higher in the group with complete return than in the group with incomplete or failed return ( P = .001 and .01, respectively). Postoperative forward elevation was significantly better in the complete return group ( P = .01)., Conclusion: This study demonstrated that 97% of elderly swimmers who underwent arthroscopic rotator cuff repair could return to swimming. The complete return rate was 56%; however, the group with incomplete or failed return showed poorer active forward elevation. Freestyle had the highest complete return rate, whereas the butterfly stroke had the lowest return rate. It may be important to achieve good active forward elevation postoperatively to return to swimming., Competing Interests: One or more of the authors has declared the following potential conflict of interest or source of funding: H.S. has received grants from Wright Medical. N.T. has received grants from Mochidaseiyaku Inc. K.M. has received grants from Exactech and Arthrex., (© The Author(s) 2020.)

Purpose: To investigate the incidence of axillary nerve palsy after arthroscopic shoulder stabilization and to measure the distance between the nerve and capsule in shoulders with a capsular lesion., Methods: This retrospective study included 2,027 shoulders (1,909 patients; 1,433 male and 476 female patients; mean age, 32 years [age range, 13-81 years]) subjected to arthroscopic soft-tissue stabilization for recurrent shoulder instability from 2005 to 2017. The exclusion criteria were bone grafting or transfer and preoperative axillary nerve symptoms. We retrospectively reviewed patient records and investigated the incidence and clinical features of axillary nerve palsy. We measured the closest distance between the axillary nerve and capsule on preoperative magnetic resonance images., Results: Postoperative axillary nerve palsy occurred in 4 shoulders (0.2% of all arthroscopic stabilizations). Capsular repair was performed in 2 shoulders (1.2% of 160 capsular repairs); humeral avulsion of the glenohumeral ligament (HAGL) repair, 1 shoulder (2% of 47 HAGL repairs); and isolated Bankart repair, 1 shoulder (0.05% of 1,941 Bankart repairs). The closest distance between the nerve and capsule was 3.4 ± 3.2 mm in shoulders with capsular or HAGL lesions and less than 1 mm in the 3 shoulders with palsy. The common symptoms in axillary nerve palsy cases were shoulder discomfort, delayed recovery of range of motion, and deltoid weakness and atrophy. A definitive diagnosis was made with electromyography in all cases. Nerve injury by a suture was confirmed during revision surgery in 3 shoulders subjected to capsular or HAGL repair during the initial operation. The palsy was transient and fully recovered in 1 shoulder with isolated Bankart repair., Conclusions: The incidence of axillary nerve palsy after arthroscopic soft-tissue shoulder stabilization was low but higher in shoulders subjected to capsular or HAGL repair. We should always consider the possibility of axillary nerve palsy in shoulders that require capsular or HAGL repair., Level of Evidence: Level IV, therapeutic case series., (Copyright © 2020 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.)

Epithelial tissues undergo cell turnover both during development and for homeostatic maintenance. Cells that are no longer needed are quickly removed without compromising the barrier function of the tissue. During metamorphosis, insects undergo developmentally programmed tissue remodeling. However, the mechanisms that regulate this rapid tissue remodeling are not precisely understood. Here, we show that the temporal dynamics of endocytosis modulate physiological cell properties to prime larval epidermal cells for cell elimination. Endocytic activity gradually reduces as tissue remodeling progresses. This reduced endocytic activity accelerates cell elimination through the regulation of Myosin II subcellular reorganization, junctional E-cadherin levels, and caspase activation. Whereas the increased Myosin II dynamics accelerates cell elimination, E-cadherin plays a protective role against cell elimination. Reduced E-cadherin is involved in the amplification of caspase activation by forming a positive-feedback loop with caspase. These findings reveal the role of endocytosis in preventing cell elimination and in the cell-property switching initiated by the temporal dynamics of endocytic activity to achieve rapid cell elimination during tissue remodeling., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2020. Published by The Company of Biologists Ltd.)

Background: Flexor digitorum superficialis (FDS) muscle provides dynamic stabilization and medial elbow support for ulnar collateral ligament (UCL). The FDS contraction significantly affects the medial joint distance (MJD) through grip contraction. However, it remains unclear whether FDS activity alone contributes to medial elbow stability, or together with the activation of the flexor digitorum profundus during grip contraction, and which finger's FDS is the main contributor to elbow stability. We investigated the resistive effects of isolated FDS contraction in individual fingers against valgus stress in the elbow joint using stress ultrasonography (US)., Methods: We investigated 17 healthy males (mean age, 27 ± 5 years). Valgus stress US was performed using the Telos device, with the elbow at 30° flexion. MJD was measured for each arm during 3 separate conditions: at rest (unloaded), under valgus load (50 N) (loaded), and under valgus load with FDS contracted in individual fingers (loaded-contracted)., Results: MJD was significantly longer when loaded (5.4 ± 0.4 mm) than unloaded (4.1 ± 0.2 mm, P = 0.007) or loaded-contracted (4.6 ± 0.3 mm, P = 0.003) for each finger. When loaded-contracted, MJD differed statistically between the index and ring fingers (P = 0.03) and between the middle and ring fingers (P = 0.04). However, the difference between the index and middle fingers was not statistically significant (P = 0.08)., Conclusions: Individual FDS contraction, particularly of the index and middle fingers, contributes most to stabilization against valgus stress. Thus, injury care programs should incorporate FDS exercises of these fingers.

The Varkud satellite ribozyme catalyses site-specific RNA cleavage and ligation, and serves as an important model system to understand RNA catalysis. Here, we combine stereospecific phosphorothioate substitution, precision nucleobase mutation and linear free-energy relationship measurements with molecular dynamics, molecular solvation theory and ab initio quantum mechanical/molecular mechanical free-energy simulations to gain insight into the catalysis. Through this confluence of theory and experiment, we unify the existing body of structural and functional data to unveil the catalytic mechanism in unprecedented detail, including the degree of proton transfer in the transition state. Further, we provide evidence for a critical Mg 2+ in the active site that interacts with the scissile phosphate and anchors the general base guanine in position for nucleophile activation. This novel role for Mg 2+ adds to the diversity of known catalytic RNA strategies and unifies functional features observed in the Varkud satellite, hairpin and hammerhead ribozyme classes.

Expanding the number of nucleotides in DNA increases the information density of functional DNA molecules, creating nanoassemblies that cannot be invaded by natural DNA/RNA in complex biological systems. Here, we show how six-letter GACTZP DNA contributes this property in two parts of a nanoassembly: 1) in an aptamer evolved from a six-letter DNA library to selectively bind liver cancer cells; and 2) in a six-letter self-assembling GACTZP nanotrain that carries the drug doxorubicin. The aptamer-nanotrain assembly, charged with doxorubicin, selectively kills liver cancer cells in culture, as the selectivity of the aptamer binding directs doxorubicin into the aptamer-targeted cells. The assembly does not kill untransformed cells that the aptamer does not bind. This architecture, built with an expanded genetic alphabet, is reminiscent of antibodies conjugated to drugs, which presumably act by this mechanism as well, but with the antibody replaced by an aptamer., (© 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Fatty degeneration of the rotator cuff muscles is reported to be associated with retear after rotator cuff repair. The purpose of this study was to assess the relationship between retear and preoperative fatty degeneration, as quantified by T2 mapping., Methods: This prospective cohort study included 83 large and 24 massive rotator cuff tears (average age, 67 years; range, 46-82 years). All patients preoperatively underwent T2 mapping magnetic resonance imaging, and T2 values of the supraspinatus and infraspinatus muscles were quantified. Cuff integrity was evaluated with magnetic resonance imaging 1 year postoperatively. Preoperative T2 values were compared between the retear and intact groups. The preoperative Goutallier stage, Constant score, and the shoulder score of the University of California at Los Angeles were also compared between the 2 groups., Results: Retear was found in 32 shoulders (30%). Postoperative Constant and University of California at Los Angeles scores were significantly higher in intact shoulders than in retear shoulders (P < .001 for both). Mean preoperative T2 values of supraspinatus and infraspinatus were 77.4 ± 13.2 ms and 73.2 ± 15.3 ms in retear shoulders and 66.5 ± 11.1 ms and 58.6 ± 11.7 ms in intact shoulders, respectively; the differences were significant in both muscles (P < .001). Cutoff values for prediction of retear were 71.8 ms in supraspinatus and 63.1 ms in infraspinatus. There were no significant differences in the preoperative Goutallier stages of supraspinatus and infraspinatus between the 2 groups., Conclusions: Retear shoulders demonstrated significantly higher preoperative T2 values than intact shoulders. T2 mapping can be a useful tool for predicting postoperative retears., (Copyright © 2019 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.)

Background: Asians generally have smaller stature than Europeans and Americans, and currently available implants used in reverse shoulder arthroplasty might not fit smaller bony anatomies. However, few articles have reported glenoid geometry in the Asian population. The purpose of this study was to measure the dimensions and orientations of the glenoid from three-dimensional computed tomography reconstructions of elderly Japanese subjects., Methods: This study included 100 shoulders (50 males and 50 females with >50 years of age). The mean age was 67 ± 7 years for both sexes, and the mean height was 167 ± 7 cm for males and 154 ± 6 cm for females. Three-dimensional scapular models were created from computed tomographic images, and the glenoid height, glenoid width, glenoid version, glenoid inclination, vault depth, and vault width were measured., Results: The mean glenoid height and width were 38.6 and 29.4 mm for males and 33.1 and 24.4 mm for females, respectively. Both retroversion and superior inclination were approximately 3° in both sexes. The glenoid vault was deeper in the posterior region with the maximum depth of 26.1 and 23.6 mm in males and females. The vault width was narrower in the anterior region with the anterior width of 2.5 mm at 15 mm medial from the glenoid face in females., Conclusion: Glenoids of Japanese females are small compared to currently available baseplates for reverse shoulder arthroplasty. These results may be helpful to aid design in smaller baseplates that better fit the anatomic geometry of the Asian glenoid., (Copyright © 2018 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

Background: Partial articular supraspinatus tendon avulsion (PASTA) lesions are often seen in shoulders of throwing athletes. PASTA lesions in anterior instability are also found in recurrent anterior glenohumeral instability., Purpose: To investigate the prevalence and location of rotator cuff tears (RCTs), including PASTA lesions, in shoulders with recurrent anterior instability., Study Design: Case series; Level of evidence, 4., Methods: A total of 842 shoulders in 801 patients with recurrent anterior instability (647 men, 154 women; mean age, 24 years; age range, 13-40 years) who underwent arthroscopic surgery at the age of 40 years or younger were enrolled. During surgery, the thickness and the sites of RCTs as well as patient factors associated with RCTs were examined., Results: RCTs were found in 57 shoulders (7%) in 56 patients. There were 4 shoulders with full-thickness RCTs and 54 shoulders with PASTA lesions. All lesions but 1 involved the anterior border of the supraspinatus (SSP). Participation in high-level athletics and older age at injury were associated with RCTs., Conclusion: The incidence of RCTs was 7% in shoulders with recurrent anterior instability at age 40 years or younger. Most RCTs were PASTA lesions that involved the anterior border of the SSP., Competing Interests: One or more of the authors has declared the following potential conflict of interest or source of funding: H.S. is a paid speaker/presenter for Wright Medical, DePuy Synthes, and Smith & Nephew. AOSSM checks author disclosures against the Open Payments Database (OPD). AOSSM has not conducted an independent investigation on the OPD and disclaims any liability or responsibility relating thereto.

To improve the management outcomes and diagnostic accuracy of the ulnar collateral ligament (UCL) injury, the anatomy of the medial side of the elbow joint is necessary to be understood in terms of the periarticular surroundings rather than the specific ligaments. The aim of this study was to anatomically clarify the medial side of the elbow joint in terms of the tendinous structures and joint capsule. We conducted a descriptive anatomical study of 23 embalmed cadaveric elbows. We macroscopically analyzed the relationship between the flexor pronator muscles (FPMs) and the joint capsule in 10 elbows, histologically analyzed in 6 elbows, and observed the bone morphology through micro computed tomography in 7 elbows. The two tendinous septa (TS) were found: between the pronator teres (PT) and flexor digitorum superficial (FDS) muscles, and between the FDS and flexor carpi ulnaris (FCU) muscles. These two TS are connected to the medial part of the brachialis tendon, deep aponeurosis of the FDS, and FCU to form the tendinous complex, which linked the humeroulnar joint and could not be histologically separated from each other. Moreover, the capsule of the humeroulnar joint under the tendinous complex had attachment on the ST of 7 mm width. The two TS, the brachialis tendon, the deep FDS and FCU aponeuroses, and the joint capsule linked the humeroulnar joint. These anatomical findings could lead to a paradigm shift in the prevention, diagnosis, and treatment of UCL injuries in baseball players. Clin. Anat. 32:379-389, 2019. © 2018 The Authors. Clinical Anatomy published by Wiley Periodicals, Inc. on behalf of American Association of Clinical Anatomists., (© 2018 The Authors. Clinical Anatomy published by Wiley Periodicals, Inc. on behalf of American Association of Clinical Anatomists.)

We report DNA- and RNA-like systems built from eight nucleotide "letters" (hence the name "hachimoji") that form four orthogonal pairs. These synthetic systems meet the structural requirements needed to support Darwinian evolution, including a polyelectrolyte backbone, predictable thermodynamic stability, and stereoregular building blocks that fit a Schrödinger aperiodic crystal. Measured thermodynamic parameters predict the stability of hachimoji duplexes, allowing hachimoji DNA to increase the information density of natural terran DNA. Three crystal structures show that the synthetic building blocks do not perturb the aperiodic crystal seen in the DNA double helix. Hachimoji DNA was then transcribed to give hachimoji RNA in the form of a functioning fluorescent hachimoji aptamer. These results expand the scope of molecular structures that might support life, including life throughout the cosmos., (Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Purpose: To evaluate the potential of an additional application of two novel hydrophobic experimental adhesive resins with or without bioactive zinc fluoride glass to promote the bond strength of a one-step self-etch universal adhesive., Materials and Methods: Three self-etch universal adhesives, G-Premio Bond (GPB), Scotchbond Universal (SBU) and Clearfil SE Bond 2 (SE2), and two experimental adhesive resins, BZF210 and BZF21, were used in this study; thus, five groups were formed: GPB, GPB+BZF210, GPB+BZF21, SBU, and SE2. The adhesives were applied to flat dentin surfaces according to each manufacturer's instructions. The microtensile bond strengths (μTBS) were evaluated after 24-h water storage. The fracture modes and interfacial structures were analyzed using SEM, while elemental analysis was performed using SEM-EDS. The data were analyzed using one-way ANOVA and the Games-Howell test (p < 0.05)., Results: Significantly higher μTBS was achieved by additional application of BZF210 (48.68 ± 6.59 MPa) and BZF21 (58.58 ± 2.84 MPa) compared with GPB (33.57 ± 4.22 MPa) alone. Most failures occurred above the smear layer in GBP, while more cohesive and mixed failures were observed in GBP+BZF210, GPB+BZF21, SBU, and SE2. The interfacial structures revealed that GBP+BZF210 and GPB+BZF21 had more and longer resin tags than did GPB. SEM-EDS showed a particularly high peak of zinc in GPB+BZF21., Conclusions: The bond strength of GPB was significantly improved by the additional application of BZF210 and BZF21. Using an additional bioactive hydrophobic layer on a one-step, self-etch universal adhesive can significantly improve its bonding efficacy and extend its clinical options.

Background: Changes over time in shoulder kinematics and function after reverse shoulder arthroplasty have not been reported. The purpose of this study was to compare shoulder kinematics and function at 6 months and 1 year after reverse shoulder arthroplasty., Methods: Twenty patients with a mean age of 74 years (range, 63-91 years) were enrolled in this study. Fluoroscopic images during scapular plane elevation were recorded at the mean of 6 months (range, 5-8 months) and 14 months (range, 11-21 months). CT-derived glenosphere models and computer-aided design humeral implant models were matched with the silhouette of the implants in the fluoroscopic images using model-image registration techniques. Glenosphere and humeral implant kinematics during scaption were compared between the two time points. Patients were also clinically examined with active range of motion and Constant score, and postoperative improvement in shoulder function were assessed., Results: Active flexion and Constant score improved after surgery (p < 0.001 for both), but there was no significant improvement after six months. There was no significant improvement in active external rotation at either postoperative exam. There were no significant differences in glenosphere or humeral kinematics between six months and one year., Interpretation: There was no significant additional improvement in either shoulder kinematics during scapular plane elevation or function between the sixth and twelfth postoperative months. We can assess kinematics at six months after reverse shoulder arthroplasty to determine how the shoulder will move. Clinically, treatment in the first six postoperative months should be emphasized to achieve better surgical outcomes., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

According to the iconic model, the Watson-Crick double helix exploits nucleobase pairs that are both size complementary (big purines pair with small pyrimidines) and hydrogen bond complementary (hydrogen bond donors pair with hydrogen bond acceptors). Using a synthetic biology strategy, we report here the discovery of two new DNA-like systems that appear to support molecular recognition with the same proficiency as standard Watson-Crick DNA. However, these both violate size complementarity (big pairs with small), retaining hydrogen bond complementarity (donors pair with acceptors) as their only specificity principle. They exclude mismatches as well as standard Watson-Crick DNA excludes mismatches. In crystal structures, these "skinny" and "fat" systems form the expected hydrogen bonds, while conferring novel minor groove properties to the resultant duplex regions of the DNA oligonucleotides. Further, computational tools, previously tested primarily on natural DNA, appear to work well for these two new molecular recognition systems, offering a validation of the power of modern computational biology. These new molecular recognition systems may have application in materials science and synthetic biology, and in developing our understanding of alternative ways that genetic information might be stored and transmitted.

The next challenge in synthetic biology is to be able to replicate synthetic nucleic acid sequences efficiently. The synthetic pair, 2-amino-8-(1-beta-d-2'- deoxyribofuranosyl) imidazo [1,2-a]-1,3,5-triazin-[8H]-4-one (trivially designated P) with 6-amino-3-(2'-deoxyribofuranosyl)-5-nitro-1H-pyridin-2-one (trivially designated Z), is replicated by certain Family A polymerases, albeit with lower efficiency. Through directed evolution, we identified a variant KlenTaq polymerase (M444V, P527A, D551E, E832V) that incorporates dZTP opposite P more efficiently than the wild-type enzyme. Here, we report two crystal structures of this variant KlenTaq, a post-incorporation complex that includes a template-primer with P:Z trapped in the active site (binary complex) and a pre-incorporation complex with dZTP paired to template P in the active site (ternary complex). In forming the ternary complex, the fingers domain exhibits a larger closure angle than in natural complexes but engages the template-primer and incoming dNTP through similar interactions. In the binary complex, although many of the interactions found in the natural complexes are retained, there is increased relative motion of the thumb domain. Collectively, our analyses suggest that it is the post-incorporation complex for unnatural substrates that presents a challenge to the natural enzyme and that more efficient replication of P:Z pairs requires a more flexible polymerase.

Described here are the synthesis, enzymology and some applications of a purine nucleoside analog (H) designed to have two tautomeric forms, one complementary to thymidine (T), the other complementary to cytidine (C). The performance of H is compared by various metrics to performances of other 'biversal' analogs that similarly rely on tautomerism to complement both pyrimidines. These include (i) the thermodynamic stability of duplexes that pair these biversals with various standard nucleotides, (ii) the ability of the biversals to support polymerase chain reaction (PCR), (iii) the ability of primers containing biversals to equally amplify targets having polymorphisms in the primer binding site, and (iv) the ability of ligation-based assays to exploit the biversals to detect medically relevant single nucleotide polymorphisms (SNPs) in sequences flanked by medically irrelevant polymorphisms. One advantage of H over the widely used inosine 'universal base' and 'mixed sequence' probes is seen in ligation-based assays to detect SNPs. The need to detect medically relevant SNPs within ambiguous sequences is especially important when probing RNA viruses, which rapidly mutate to create drug resistance, but also suffer neutral drift, the second obstructing simple methods to detect the first. Thus, H is being developed to detect variants of viruses that are rapidly mutating.

Purpose: To determine the microtensile bond strength (µTBS) of two 2-step self-etch adhesives (SEAs) to bur-cut and uncut enamel., Materials and Methods: The buccal and lingual enamel surfaces of 15 teeth were ground flat ("bur-cut" enamel), while the enamel surface of another set of 15 teeth was solely prophylactically cleaned ("uncut" enamel). Resin composite was bonded to the surfaces using the GPDM-based SEA OptiBond XTR (Kerr), the 10-MDP-based SEA Clearfil SE Bond (Kuraray Noritake), or the 3-step etch-and-rinse adhesive (E&RA) OptiBond FL (Kerr) that served as the gold-standard control. After 1-week water storage at 37°C, specimens were serially cut into 1-mm2 stick-shaped microspecimens, of which half per tooth were further subjected to 20,000 thermocycles (TC) prior to µTBS testing. Data were statistically analyzed using ANOVA and the post-hoc Tukey test. The interfacial ultrastructure of the GPDM-based SEA OptiBond XTR with uncut and bur-cut enamel was additionally characterized with transmission electron microscopy (TEM)., Results: After 1-week water storage and upon TC aging, both SEAs bonded significantly (p < 0.05) less effectively to both bur-cut and uncut enamel than did the control OptiBond FL, except when OptiBond XTR was bonded to bur-cut enamel; then a similarly high µTBS was recorded (p > 0.05). TEM interfacial characterization revealed a tight interaction of Optibond XTR with both bur-cut and uncut enamel., Conclusion: The best bonding efficacy to enamel is still achieved by etching with phosphoric acid, following an E&R approach, while the higher etching efficacy of the GPDM-based SEA may result in equally effective bonding, provided that the enamel is bur-roughened first.

Background: Capitellar osteochondritis dissecans (OCD) in skeletally immature athletes has often been seen in baseball players and gymnasts. The choice of surgical procedure for unstable lesions in skeletally immature athletes remains controversial., Purpose: To investigate functional outcomes and radiographic changes in the midterm to long-term postoperative period after arthroscopic (AS) resection for small to large capitellar OCD lesions in skeletally immature athletes., Study Design: Case series; Level of evidence, 4., Methods: A total of 38 elbows in 38 patients (33 boys, 5 girls; mean age, 14 years [range, 13-15 years]) with skeletally immature elbows underwent AS resection for capitellar OCD. Patients were observed for at least 5 years (mean, 8 years [range, 5-12 years]). Elbows with a lesion width that did not exceed one-half of the radial head diameter were assigned to group 1 (n = 17 elbows), and larger lesions were assigned to group 2 (n = 21 elbows). Functional scores, patient satisfaction, range of motion (ROM), and osteoarthritis (OA) grades were evaluated between the groups., Results: All patients returned to sports activity. Functional scores at the final follow-up were not significantly different between the groups. Patient satisfaction scores were significantly higher in group 1 than in group 2. There was significant improvement in flexion ROM at the final follow-up compared with preoperative values in group 1 ( P = .017), and there was a significant between-group difference (group 1: 141°; group 2: 133°; P = .002). Extension ROM showed significant improvement in both groups (group 1: from -8° to 3°; group 2: from -17° to -1°; P < .001 for both). Group 1 tended to have better extension than group 2, but the difference was not significant. There were no elbows with severe OA in either group, but the OA grade progressed in 5 elbows (29%) in group 1 and 9 elbows (43%) in group 2, and this rate of OA progression was statistically significant between groups ( P = .005)., Conclusion: Both functional outcomes and radiological findings after AS fragment resection were excellent in elbows with small lesions. Although overall outcomes were acceptable in elbows with larger lesions, flexion ROM and patient satisfaction scores were significantly inferior to those in elbows with smaller lesions., Competing Interests: The authors declared that they have no conflicts of interest in the authorship and publication of this contribution.

A goal of synthetic biology is to develop new nucleobases that retain the desirable properties of natural nucleobases at the same time as expanding the genetic alphabet. The nonstandard Watson-Crick pair between imidazo[1,2-a]-1,3,5-triazine-2(8H)-4(3H)-dione (X) and 2,4-diaminopyrimidine (K) does exactly this, pairing via complementary arrangements of hydrogen bonding in these two nucleobases, which do not complement any natural nucleobase. Here, we report the crystal structure of a duplex DNA oligonucleotide in B-form including two consecutive X:K pairs in GATCXK DNA determined as a host-guest complex at 1.75 Å resolution. X:K pairs have significant propeller twist angles, similar to those observed for A:T pairs, and a calculated hydrogen bonding pairing energy that is weaker than that of A:T. Thus, although inclusion of X:K pairs results in a duplex DNA structure that is globally similar to that of an analogous G:C structure, the X:K pairs locally and energetically more closely resemble A:T pairs.

Synthetic nucleobases presenting non-Watson-Crick arrangements of hydrogen bond donor and acceptor groups can form additional nucleotide pairs that stabilize duplex DNA independent of the standard A:T and G:C pairs. The pair between 2-amino-3-nitropyridin-6-one 2'-deoxyriboside (presenting a {donor-donor-acceptor} hydrogen bonding pattern on the Watson-Crick face of the small component, trivially designated Z) and imidazo[1,2-a]-1,3,5-triazin-4(8H)one 2'-deoxyriboside (presenting an {acceptor-acceptor-donor} hydrogen bonding pattern on the large component, trivially designated P) is one of these extra pairs for which a substantial amount of molecular biology has been developed. Here, we report the results of UV absorbance melting measurements and determine the energetics of binding of DNA strands containing Z and P to give short duplexes containing Z:P pairs as well as various mismatches comprising Z and P. All measurements were done at 1 M NaCl in buffer (10 mM Na cacodylate, 0.5 mM EDTA, pH 7.0). Thermodynamic parameters (ΔH°, ΔS°, and ΔG° 37 ) for oligonucleotide hybridization were extracted. Consistent with the Watson-Crick model that considers both geometric and hydrogen bonding complementarity, the Z:P pair was found to contribute more to duplex stability than any mismatches involving either nonstandard nucleotide. Further, the Z:P pair is more stable than a C:G pair. The Z:G pair was found to be the most stable mismatch, forming either a deprotonated mismatched pair or a wobble base pair analogous to the stable T:G mismatch. The C:P pair is less stable, perhaps analogous to the wobble pair observed for C:O 6 -methyl-G, in which the pyrimidine is displaced into the minor groove. The Z:A and T:P mismatches are much less stable. Parameters for predicting the thermodynamics of oligonucleotides containing Z and P bases are provided. This represents the first case where this has been done for a synthetic genetic system.

The implants used in reverse shoulder arthroplasty were originally designed for white patients; thus, they might not be optimal for smaller Asian patients. The selection of reverse shoulder arthroplasty systems is limited in Japan. The purpose of this study was to measure the geometry of the proximal humerus in elderly Japanese patients to inform implant selection according to humerus size. This study included 155 shoulders from 148 patients 50 years or older who underwent computed tomography for shoulder disorders other than arthritis and trauma. There were 67 male and 81 female patients with mean ages of 68 and 66 years, respectively. The humeral head diameter, head height, neck-shaft angle, neck diameters, and osteotomy diameters for 155° inclination implants were measured using surface models created from computed tomography scans. The mean diameter of head curvature, humeral head height, and neck-shaft angle were 48.2 mm, 19.8 mm, and 136° for male patients and 42.4 mm, 17.1 mm, and 136° for female patients, respectively. The mean mediolateral and anteroposterior neck diameters were 46.9 mm and 43.7 mm for men and 41.6 mm and 38.4 mm for women, respectively. The mean mediolateral and anteroposterior osteotomy diameters were 42.2 mm and 41.2 mm for male patients and 38.6 mm and 36.7 mm for female patients, respectively. There were significant differences between the sexes in all measurements except neck-shaft angle. Humeral implants with 155° inclination will not fit the humerus of smaller Japanese women. Thus, implants with anatomical inclination and an onlay humeral tray may be a better choice for smaller patients. [Orthopedics. 2017; 40(3):e485-e490.]., (Copyright 2017, SLACK Incorporated.)

Background: Fatty degeneration of the cuff muscles is usually evaluated at the Y-view in oblique sagittal images. It was recently proposed that muscle shift after repair may influence the fatty degeneration values, and the evaluation of the muscles at a more medial site was recommended. However, the differences in muscle quality in accord with measurement sites have been unclear. Here we evaluated differences in fatty degeneration of the rotator cuff muscles measured quantitatively at different sites, using T2 mapping., Methods: We assessed 702 shoulders of 675 patients (335 males, 340 females; mean age, 62 years) who underwent MRI including T2 mapping. There were 345 shoulders without rotator cuff tears and 357 shoulders with tears: partial tear = 103 shoulders; small = 63; medium = 94; large = 71; massive = 26. T2 values of the supraspinatus and infraspinatus muscles were measured on the Y-view and on the image that was 15 mm medial to the Y-view., Results: The T2 values at the medial site increased with the tear extent, as did those on the Y-view. There were no significant differences in supraspinatus T2 values between those on the Y-view and at the medial site in all tear size groups except medium and large tears (p = 0.008 and p < 0.001, respectively). There were also no significant differences in infraspinatus T2 values between the two sites in all tear size groups except large tears (p = 0.002). However, the differences were relatively small (2.4-5.6 ms), which were within the standard deviations of the measurements., Conclusions: The T2 values of the supraspinatus and infraspinatus muscles on the Y-view and at 15 mm medial to it were almost identical, with the exception of small differences in the case of larger tears., (Copyright © 2016. Published by Elsevier B.V.)

Background: This study investigated the morphologic changes in the biceps tendon using ultrasonography before and after successful arthroscopic posterosuperior rotator cuff repair., Methods: Forty-four patients (44 shoulders) underwent arthroscopic posterosuperior rotator cuff repair with 1-year postoperative magnetic resonance imaging (MRI) follow-up. The patients comprised 22 men and 22 women with an average age of 61 years. The cross-sectional area (CSA) of the biceps tendon in the bicipital groove was measured, and the vascularity in the bicipital groove was graded as 0 to 3, based on the signal density of the anterior circumflex artery, using power Doppler ultrasonography. The preoperative and postoperative CSA and vascularity grades were compared. The pain score on the University of California, Los Angeles Shoulder Rating Scale was used to analyze the correlation between vascularity and postoperative pain., Results: The average preoperative and postoperative CSA of the biceps tendon was 15.4 ± 6.5 and 17.9 ± 7.5 mm 2 , respectively. The postoperative CSA was significantly larger than the preoperative CSA (P < .01). Although no significant difference in the vascularity of the bicipital groove was observed between preoperative and postoperative grading, a negative correlation was observed between the vascularity and postoperative pain score on the University of California, Los Angeles scale (r = -0.369)., Conclusions: The biceps tendon in the bicipital groove becomes thicker over time, even after successful posterosuperior rotator cuff repair. In addition, an increase in the vascularity around the biceps tendon in the groove is correlated with pain symptoms after successful repair., (Copyright © 2017 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.)

Reported here is a laboratory in vitro evolution (LIVE) experiment based on an artificially expanded genetic information system (AEGIS). This experiment delivers the first example of an AEGIS aptamer that binds to an isolated protein target, the first whose structural contact with its target has been outlined and the first to inhibit biologically important activities of its target, the protective antigen from Bacillus anthracis We show how rational design based on secondary structure predictions can also direct the use of AEGIS to improve the stability and binding of the aptamer to its target. The final aptamer has a dissociation constant of ∼35 nM. These results illustrate the value of AEGIS-LIVE for those seeking to obtain receptors and ligands without the complexities of medicinal chemistry, and also challenge the biophysical community to develop new tools to analyze the spectroscopic signatures of new DNA folds that will emerge in synthetic genetic systems replacing standard DNA and RNA as platforms for LIVE., (© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.)

In its "grand challenge" format in chemistry, "synthesis" as an activity sets out a goal that is substantially beyond current theoretical and technological capabilities. In pursuit of this goal, scientists are forced across uncharted territory, where they must answer unscripted questions and solve unscripted problems, creating new theories and new technologies in ways that would not be created by hypothesis-directed research. Thus, synthesis drives discovery and paradigm changes in ways that analysis cannot. Described here are the products that have arisen so far through the pursuit of one grand challenge in synthetic biology: Recreate the genetics, catalysis, evolution, and adaptation that we value in life, but using genetic and catalytic biopolymers different from those that have been delivered to us by natural history on Earth. The outcomes in technology include new diagnostic tools that have helped personalize the care of hundreds of thousands of patients worldwide. In science, the effort has generated a fundamentally different view of DNA, RNA, and how they work., (Copyright © 2016 Cold Spring Harbor Laboratory Press; all rights reserved.)

Noroviruses are the major cause of global viral gastroenteritis with short incubation times and small inoculums required for infection. This creates a need for a rapid molecular test for norovirus for early diagnosis, in the hope of preventing the spread of the disease. Non-chemists generally use off-the shelf reagents and natural DNA to create such tests, suffering from background noise that comes from adventitious DNA and RNA (collectively xNA) that is abundant in real biological samples, especially feces, a common location for norovirus. Here, we create an assay that combines artificially expanded genetic information systems (AEGIS, which adds nucleotides to the four in standard xNA, pairing orthogonally to A:T and G:C) with loop-mediated isothermal amplification (LAMP) to amplify norovirus RNA at constant temperatures, without the power or instrument requirements of PCR cycling. This assay was then validated using feces contaminated with murine norovirus (MNV). Treating stool samples with ammonia extracts the MNV RNA, which is then amplified in an AEGIS-RT-LAMP where AEGIS segments are incorporated both into an internal LAMP primer and into a molecular beacon stem, the second lowering background signaling noise. This is coupled with RNase H nicking during sample amplification, allowing detection of as few as 10 copies of noroviral RNA in a stool sample, generating a fluorescent signal visible to human eye, all in a closed reaction vessel., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This paper combines two advances to detect MERS-CoV, the causative agent of Middle East Respiratory Syndrome, that have emerged over the past few years from the new field of "synthetic biology". Both are based on an older concept, where molecular beacons are used as the downstream detection of viral RNA in biological mixtures followed by reverse transcription PCR amplification. The first advance exploits the artificially expanded genetic information systems (AEGIS). AEGIS adds nucleotides to the four found in standard DNA and RNA (xNA); AEGIS nucleotides pair orthogonally to the A:T and G:C pairs. Placing AEGIS components in the stems of molecular beacons is shown to lower noise by preventing unwanted stem invasion by adventitious natural xNA. This should improve the signal-to-noise ratio of molecular beacons operating in complex biological mixtures. The second advance introduces a nicking enzyme that allows a single target molecule to activate more than one beacon, allowing "signal amplification". Combining these technologies in primers with components of a self-avoiding molecular recognition system (SAMRS), we detect 50 copies of MERS-CoV RNA in a multiplexed respiratory virus panel by generating fluorescence signal visible to human eye and/or camera., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Laboratory in vitro evolution (LIVE) might deliver DNA aptamers that bind proteins expressed on the surface of cells. In this work, we used cell engineering to place glypican 3 (GPC3), a possible marker for liver cancer theranostics, on the surface of a liver cell line. Libraries were then built from a six-letter genetic alphabet containing the standard nucleobases and two added nucleobases (2-amino-8H-imidazo[1,2-a][1,3,5]triazin-4-one and 6-amino-5-nitropyridin-2-one), Watson-Crick complements from an artificially expanded genetic information system (AEGIS). With counterselection against non-engineered cells, eight AEGIS-containing aptamers were recovered. Five bound selectively to GPC3-overexpressing cells. This selection-counterselection scheme had acceptable statistics, notwithstanding the possibility that cells engineered to overexpress GPC3 might also express different off-target proteins. This is the first example of such a combination., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

In addition to completing the Watson-Crick nucleobase matching "concept" (big pairs with small, hydrogen bond donors pair with hydrogen bond acceptors), artificially expanded genetic information systems (AEGIS) also challenge DNA polymerases with a complete set of mismatches, including wobble mismatches. Here, we explore wobble mismatches with AEGIS with DNA polymerase 1 from Escherichia coli. Remarkably, we find that the polymerase tolerates an AEGIS:standard wobble that has the same geometry as the G:T wobble that polymerases have evolved to exclude but excludes a wobble geometry that polymerases have never encountered in natural history. These results suggest certain limits to "structural analogy" and "evolutionary guidance" as tools to help synthetic biologists expand DNA alphabets.

One frontier in synthetic biology seeks to move artificially expanded genetic information systems (AEGIS) into natural living cells and to arrange the metabolism of those cells to allow them to replicate plasmids built from these unnatural genetic systems. In addition to requiring polymerases that replicate AEGIS oligonucleotides, such cells require metabolic pathways that biosynthesize the triphosphates of AEGIS nucleosides, the substrates for those polymerases. Such pathways generally require nucleoside and nucleotide kinases to phosphorylate AEGIS nucleosides and nucleotides on the path to these triphosphates. Thus, constructing such pathways focuses on engineering natural nucleoside and nucleotide kinases, which often do not accept the unnatural AEGIS biosynthetic intermediates. This, in turn, requires assays that allow the enzyme engineer to follow the kinase reaction, assays that are easily confused by ATPase and other spurious activities that might arise through "site-directed damage" of the natural kinases being engineered. This article introduces three assays that can detect the formation of both natural and unnatural deoxyribonucleoside triphosphates, assessing their value as polymerase substrates at the same time as monitoring the progress of kinase engineering. Here, we focus on two complementary AEGIS nucleoside diphosphates, 6-amino-5-nitro-3-(1'-β-D-2'-deoxyribofuranosyl)-2(1H)-pyridone and 2-amino-8-(1'-β-D-2'-deoxyribofuranosyl)-imidazo[1,2-a]-1,3,5-triazin-4(8H)-one. These assays provide new ways to detect the formation of unnatural deoxyribonucleoside triphosphates in vitro and to confirm their incorporation into DNA. Thus, these assays can be used with other unnatural nucleotides.

The purpose of this study was to compare the micro-tensile bond strength (µTBS) of three resin core composites to dentin and to examine the bonded interface of the composites. One experimental TDK-03(TD) and, two commercial, DC core Automix One (DC) and Unifil core EM(UN) were used. Flat dentin surfaces of human molars were exposed using #600 SiC paper and bonded with the respective adhesive of each system. After bonding, the composites were built up on the surfaces and cured under two conditions: "light condition" or "dark condition". µTBSs (MPa) in the light condition were: TD; 60.02±17.08, DC; 38.21±13.70, and UN; 29.50±9.71; in the dark condition: TD; 54.62±17.11, DC; 8.40±4.81, and UN; 9.47±6.56. Dark curing negatively affected the bond strength of the two commercial resin-core materials. The experimental material was not affected by the curing conditions.

The purpose of this study was to evaluate the µTBS (microtensile bond strength) of currently available self-etching adhesives with an experimental self-etch adhesive in normal and caries-affected dentin, using a portable hardness measuring device, in order to standardize dentin Knoop hardness. Normal (ND) and caries-affected dentin (CAD) were obtained from twenty human molars with class II natural caries. The following adhesive systems were tested: Mega Bond (MB), a 2-step self-etching adhesive; MTB-200 (MTB), an experimental 1-step self-etching adhesive (1-SEA), and two commercially available one-step self-etching systems, G-Bond Plus (GB) and Adper Easy Bond (EB). MB-ND achieved the highest µTBS (p<0.05). The mean µTBS was statistically lower in CAD than in ND for all adhesives tested (p<0.05), and the 2-step self-etch adhesive achieved better overall performance than the 1-step self-etch adhesives.

Background: Arthroscopic osseous Bankart repair for shoulders with chronic recurrent anterior instability has been reported as an effective procedure with promising short-term outcomes. However, to date, we know of no report describing longer-term outcomes and glenoid morphologic change. The purpose of the present study was to report intermediate to long-term outcomes and glenoid morphologic change after arthroscopic osseous Bankart repair in patients with substantial glenoid bone loss., Methods: A consecutive series of eighty-five patients with traumatic anterior glenohumeral instability associated with a chronic osseous Bankart lesion underwent arthroscopic repair from January 2005 through December 2006. Forty-six patients with bone loss of >15% of the inferior glenoid diameter relative to the assumed inferior circle regardless of the fragment size were selected as candidates for this study. Thirty-eight patients (83%), including thirty-four male and four female patients, with a mean age of 23.4 years (range, fifteen to thirty-six years) at the time of surgery, were available for final follow-up at a mean of 6.2 years (range, 5.0 to 8.1 years) after surgery., Results: One patient had a redislocation during a traffic accident five months after surgery before obtaining an osseous union. The mean Rowe score and the mean Western Ontario Shoulder Instability Index improved significantly from 30.7 points preoperatively to 95.4 points postoperatively and from 26.5% to 81.5%, respectively. Although the mean preoperative fragment size was measured as only 4.7%, the mean glenoid bone loss improved from 20.4% preoperatively to -1.1% postoperatively., Conclusions: Arthroscopic osseous Bankart repair is an effective primary treatment for shoulders with substantial glenoid bone loss as it provides successful outcomes without recurrence of instability once osseous union is obtained. Glenoid morphology can be normalized during the intermediate to long-term postoperative period, even in shoulders with a smaller fragment., (Copyright © 2015 by The Journal of Bone and Joint Surgery, Incorporated.)

Nucleic acid (NA)-targeted tests detect and quantify viral DNA and RNA (collectively xNA) to support epidemiological surveillance and, in individual patients, to guide therapy. They commonly use polymerase chain reaction (PCR) and reverse transcription PCR. Although these all have rapid turnaround, they are expensive to run. Multiplexing would allow their cost to be spread over multiple targets, but often only with lower sensitivity and accuracy, noise, false positives, and false negatives; these arise by interactions between the multiple nucleic acid primers and probes in a multiplexed kit. Here we offer a multiplexed assay for a panel of respiratory viruses that mitigates these problems by combining several nucleic acid analogs from the emerging field of synthetic biology: (i) self-avoiding molecular recognition systems (SAMRSs), which facilitate multiplexing, and (ii) artificially expanded genetic information systems (AEGISs), which enable low-noise PCR. These are supplemented by "transliteration" technology, which converts standard nucleotides in a target to AEGIS nucleotides in a product, improving hybridization. The combination supports a multiplexed Luminex-based respiratory panel that potentially differentiates influenza viruses A and B, respiratory syncytial virus, severe acute respiratory syndrome coronavirus (SARS), and Middle East respiratory syndrome (MERS) coronavirus, detecting as few as 10 MERS virions in a 20-μl sample., (Copyright © 2015 Elsevier Inc. All rights reserved.)

As one of its goals, synthetic biology seeks to increase the number of building blocks in nucleic acids. While efforts towards this goal are well advanced for DNA, they have hardly begun for RNA. Herein, we present a crystal structure for an RNA riboswitch where a stem C:G pair has been replaced by a pair between two components of an artificially expanded genetic-information system (AEGIS), Z and P, (6-amino-5-nitro-2(1H)-pyridone and 2-amino-imidazo[1,2-a]-1,3,5-triazin-4-(8H)-one). The structure shows that the Z:P pair does not greatly change the conformation of the RNA molecule nor the details of its interaction with a hypoxanthine ligand. This was confirmed in solution by in-line probing, which also measured a 3.7 nM affinity of the riboswitch for guanine. These data show that the Z:P pair mimics the natural Watson-Crick geometry in RNA in the first example of a crystal structure of an RNA molecule that contains an orthogonal added nucleobase pair., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Idiopathic adhesive capsulitis is defined as a frozen shoulder with severe and global range-of-motion loss of unknown etiology. The purpose of our study was to clarify the prevalence of rotator cuff lesions according to patterns and severity of range-of-motion loss in a large cohort of patients with stiff shoulders., Methods: Rotator cuff pathology was prospectively investigated with use of magnetic resonance imaging (MRI) or ultrasonography in a series of 379 stiff shoulders; patients with traumatic etiology, diabetes, or radiographic abnormalities were excluded. Eighty-nine shoulders demonstrated severe and global loss of passive motion (≤100° of forward flexion, ≤10° of external rotation with the arm at the side, and internal rotation not more cephalad than the L5 level) and were classified as having severe and global loss of motion (Group 1). The remaining 290 shoulders were divided into two groups: those with severe but not global loss (Group 2; 111 shoulders) and mild to moderate limitation (Group 3; 179 shoulders)., Results: Among all shoulders, imaging demonstrated an intact rotator cuff in 51%, a full-thickness tear in 35%, and a partial-thickness tear in 15%. In Group 1, 91% had an intact rotator cuff and 9% had a partial-thickness rotator cuff tear. No patient in this group demonstrated a full-thickness tear. In Group 2 and Group 3, respectively, 44% and 35% of the shoulders were intact, 17% and 16% had a partial-thickness tear, and 39% and 50% had a full-thickness tear., Conclusions: Shoulder stiffness with severe and global loss of passive range of motion is not associated with full-thickness rotator cuff tears, although some patients may have a partial-thickness tear. Shoulders with severe and global loss of range of motion at a first visit are likely to be cases of idiopathic adhesive capsulitis and may not require further imaging studies., (Copyright © 2015 by The Journal of Bone and Joint Surgery, Incorporated.)

Assays that detect DNA or RNA (xNA) are highly sensitive, as small amounts of xNA can be amplified by PCR. Unfortunately, PCR is inconvenient in low-resource environments, and requires equipment and power that might not be available in these environments. Isothermal procedures, which avoid thermal cycling, are often confounded by primer dimers, off-target priming, and other artifacts. Here, we show how a "self avoiding molecular recognition system" (SAMRS) eliminates these artifacts and gives clean amplicons in a helicase-dependent isothermal amplification (SAMRS-HDA). We also show that incorporating SAMRS into the 3'-ends of primers facilitates the design and screening of primers for HDA assays. Finally, we show that SAMRS-HDA can be twofold multiplexed, difficult to achieve with HDA using standard primers. Thus, SAMRS-HDA is a more versatile approach than standard HDA, with a broader applicability for xNA-targeted diagnostics and research., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Axiomatically, the density of information stored in DNA, with just four nucleotides (GACT), is higher than in a binary code, but less than it might be if synthetic biologists succeed in adding independently replicating nucleotides to genetic systems. Such addition could also add functional groups not found in natural DNA, but useful for molecular performance. Here, we consider two new nucleotides (Z and P, 6-amino-5-nitro-3-(1'-β-D-2'-deoxyribo-furanosyl)-2(1H)-pyridone and 2-amino-8-(1'-β-D-2'-deoxyribofuranosyl)-imidazo[1,2-a]-1,3,5-triazin-4(8H)-one). These are designed to pair via complete Watson-Crick geometry. These were added to a library of oligonucleotides used in a laboratory in vitro evolution (LIVE) experiment; the GACTZP library was challenged to deliver molecules that bind selectively to liver cancer cells, but not to untransformed liver cells. Unlike in classical in vitro selection, low levels of mutation allow this system to evolve to create binding molecules not necessarily present in the original library. Over a dozen binding species were recovered. The best had Z and/or P in their sequences. Several had multiple, nearby, and adjacent Zs and Ps. Only the weaker binders contained no Z or P at all. This suggests that this system explored much of the sequence space available to this genetic system and that GACTZP libraries are richer reservoirs of functionality than standard libraries.

Expanded genetic systems are most likely to work with natural enzymes if the added nucleotides pair with geometries that are similar to those displayed by standard duplex DNA. Here, we present crystal structures of 16-mer duplexes showing this to be the case with two nonstandard nucleobases (Z, 6-amino-5-nitro-2(1H)-pyridone and P, 2-amino-imidazo[1,2-a]-1,3,5-triazin-4(8H)one) that were designed to form a Z:P pair with a standard "edge on" Watson-Crick geometry, but joined by rearranged hydrogen bond donor and acceptor groups. One duplex, with four Z:P pairs, was crystallized with a reverse transcriptase host and adopts primarily a B-form. Another contained six consecutive Z:P pairs; it crystallized without a host in an A-form. In both structures, Z:P pairs fit canonical nucleobase hydrogen-bonding parameters and known DNA helical forms. Unique features include stacking of the nitro group on Z with the adjacent nucleobase ring in the A-form duplex. In both B- and A-duplexes, major groove widths for the Z:P pairs are approximately 1 Å wider than those of comparable G:C pairs, perhaps to accommodate the large nitro group on Z. Otherwise, ZP-rich DNA had many of the same properties as CG-rich DNA, a conclusion supported by circular dichroism studies in solution. The ability of standard duplexes to accommodate multiple and consecutive Z:P pairs is consistent with the ability of natural polymerases to biosynthesize those pairs. This, in turn, implies that the GACTZP synthetic genetic system can explore the entire expanded sequence space that additional nucleotides create, a major step forward in this area of synthetic biology.

Expanding the synthetic biology of artificially expanded genetic information systems (AEGIS) requires tools to make and analyze RNA molecules having added nucleotide "letters". We report here the development of T7 RNA polymerase and reverse transcriptase to catalyze transcription and reverse transcription of xNA (DNA or RNA) having two complementary AEGIS nucleobases, 6-amino-5-nitropyridin-2-one (trivially, Z) and 2-aminoimidazo[1,2a]-1,3,5-triazin-4(8H)-one (trivially, P). We also report MALDI mass spectrometry and HPLC-based analyses for oligomeric GACUZP six-letter RNA and the use of ribonuclease (RNase) A and T1 RNase as enzymatic tools for the sequence-specific degradation of GACUZP RNA. We then applied these tools to analyze the GACUZP and GACTZP products of polymerases and reverse transcriptases (respectively) made from DNA and RNA templates. In addition to advancing this 6-letter AEGIS toward the biosynthesis of proteins containing additional amino acids, these experiments provided new insights into the biophysics of DNA.

Mosquito-borne arboviruses are emerging world-wide as important human and animal pathogens. This makes assays for their accurate and rapid identification essential for public health, epidemiological, ecological studies. Over the past decade, many mono- and multiplexed assays targeting arboviruses nucleic acids have been reported. None has become established for the routine identification of multiple viruses in a "single tube" setting. With increasing multiplexing, the detection of viral RNAs is complicated by noise, false positives and negatives. In this study, an assay was developed that avoids these problems by combining two new kinds of nucleic acids emerging from the field of synthetic biology. The first is a "self-avoiding molecular recognition system" (SAMRS), which enables high levels of multiplexing. The second is an "artificially expanded genetic information system" (AEGIS), which enables clean PCR amplification in nested PCR formats. A conversion technology was used to place AEGIS component into amplicon, improving their efficiency of hybridization on Luminex beads. When Luminex "liquid microarrays" are exploited for downstream detection, this combination supports single-tube PCR amplification assays that can identify 22 mosquito-borne RNA viruses from the genera Flavivirus, Alphavirus, Orthobunyavirus. The assay differentiates between closely-related viruses, as dengue, West Nile, Japanese encephalitis, and the California serological group. The performance and the sensitivity of the assay were evaluated with dengue viruses and infected mosquitoes; as few as 6-10 dengue virions can be detected in a single mosquito., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The purpose of this study was to evaluate the microtensile bond strength (µTBS) of a newly developed resin cement, ECD-89 (ECD, Tokuyama Dental, Tokyo, Japan) to dentin and to observe the interfacial micromorphology by comparing with two commercial resin cements, Multilink Automix (MA, Ivoclar Vivadent AG, Schaan, Liechtenstein) and Panavia F2.0 (PF, Kuraray Noritake Dental, Tokyo, Japan). Flat dentin surfaces of human third molars were exposed using #600 SiC. After application of primer and cement to the dentin surface, each cement was applied and cured with light (light condition) or without light (dark condition). The teeth were sectioned to obtain beams (1 mm×1 mm) after 24 h of water storage. The mean bond strengths and SDs (MPa) were: ECD: 68.6±14.9, MA: 39.2±18.9, PF: 39.4±18.5 and ECD: 54.5±22.4, MA: 36.7±15.6, PF: 13.4±4.46 when cured in light and dark condition, respectively. In both conditions, ECD-89 showed statistically higher µTBS than the others.

The purpose of this study was to evaluate the effect of remaining dentin thickness (RDT) on the bond strength of current adhesive systems. Third molars were randomly allocated among four groups depending on the adhesive system used: Clearfil SE Bond ONE (SE1), G-Bond PLUS (GB), BeautiBond (BB), and Clearfil Mega Bond (MB). Bonded specimens were stored in water at 37°C for 24 h. Teeth were then sectioned perpendicular to the adhesive interface to produce beams. After measuring RDT of each beam, microtensile bond strength test was carried out using a universal testing machine at a crosshead speed of 1 mm/min. All data were analyzed by linear regression analysis. Bond strengths of one-step self-etch materials used in this study increased with an increase in RDT. In contrast, that of two-step self-etch adhesive system was not affected by RDT.

Recombinase polymerase amplification (RPA) is an isothermal method to amplify nucleic acid sequences without the temperature cycling that classical PCR uses. Instead of using heat to denature the DNA duplex, RPA uses recombination enzymes to swap single-stranded primers into the duplex DNA product; these are then extended using a strand-displacing polymerase to complete the cycle. Because RPA runs at low temperatures, it never forces the system to recreate base-pairs following Watson-Crick rules, and therefore it produces undesired products that impede the amplification of the desired product, complicating downstream analysis. Herein, we show that most of these undesired side products can be avoided if the primers contain components of a self-avoiding molecular recognition system (SAMRS). Given the precision that is necessary in the recombination systems for them to function biologically, it is surprising that they accept SAMRS. SAMRS-RPA is expected to be a powerful tool within the range of amplification techniques available to scientists., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)