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Purpose: The radius bone has a slight dorsoradial bow that allows for full forearm pronosupination around the ulna. However, radial malunion can lead to reversal of the radial bow and subsequent volar instability of the distal radioulnar joint (DRUJ), predominantly in supination. This study assessed the outcomes of corrective radial osteotomy for volar DRUJ instability after radial malunion in children., Methods: The charts of 7 children (2 boys and 5 girls) treated with corrective radial osteotomy for volar DRUJ instability after a radius fracture or deformity were reviewed. Demographic, diagnostic, treatment, and complication information was collected for each patient. Radiographs at initial injury, fracture union, diagnosis of DRUJ instability, and final follow-up were reviewed for radiographic measurements of radial deformity and subsequent correction., Results: Fractures included 4 distal radius, 2 proximal radius, and 1 plastic deformation of the radial shaft. Volar DRUJ instability was diagnosed an average of 2.7 years (range, 1-6 years) after fracture at an average age of 13.6 years (range, 12-17 years). Two of 7 patients had persistent symptoms despite having undergone previous soft tissue surgery for DRUJ instability. Radial osteotomy was performed on all patients (3 dorsal and 4 volar approaches), with an average sagittal plane correction of 23° ± 10° (range, 14° to 40°). Osteotomy site varied (3 proximal third, 1 middle third, and 3 distal third) based on the apex of maximal deformity. Patients were observed an average of 2.3 years (range, 1.0-5.7 years). At final follow-up, all patients had a stable DRUJ and no patient required soft tissue stabilization., Conclusions: Apex volar malunion of radial fractures may result in volar instability of the DRUJ. Radial osteotomy restored the normal apex dorsal radial bow and effectively stabilized the DRUJ without the need for soft tissue repair. Osteotomy should be tailored to the specific site of radiographic deformity., Type of Study/level of Evidence: Therapeutic V., (Copyright © 2018 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.)

Background and Objectives: Pediatric hospital-acquired venous thromboembolism (VTE) is costly, has high morbidity, and is often preventable. The objective of this quality-improvement effort was to increase the percentage of general surgery and orthopedic patients ≥10 years of age screened for VTE risk from 0% to 80%., Methods: At a freestanding children's hospital, 2 teams worked to implement VTE risk screening for postoperative inpatients. The general surgery team used residents and nurse practitioners to perform screening whereas the orthopedic team initially used bedside nursing staff. Both groups employed multiple small tests of change. Shared key interventions included refinement of a screening tool, provider education, mitigation of failures, and embedding the risk assessment task into staff workflow. The primary outcome measure, the percentage of eligible patients with a completed VTE risk assessment, was plotted on run charts. Secondary outcome measures for screened patients included the level of risk, the use of appropriate prophylaxis, and VTE events., Results: Median weekly percentage of general surgery patients screened for VTE risk increased from 0% to 86% within 12 months, and median weekly percentage of orthopedic patients screened for VTE risk increased from 0% to 46% within 8 months. Among screened patients, the majority were at low or moderate risk for VTE and received prophylaxis in accordance with or beyond guideline recommendations. No screened patients developed VTE., Conclusions: Quality-improvement methods were used to implement a VTE risk screening process for postoperative patients. Using providers as screeners, as opposed to bedside nurses, led to a greater percentage of patients screened., Competing Interests: POTENTIAL CONFLICT OF INTEREST: The authors have indicated they have no potential conflicts of interest to disclose., (Copyright © 2017 by the American Academy of Pediatrics.)

Study Design: Comparative analysis of 2 groups of patients who underwent surgical treatment of adolescent idiopathic scoliosis (AIS)., Objective: To compare a segmental pedicle screw only system with a hybrid system for the treatment of Lenke type 1 AIS curves., Summary of Background Data: Although previous AIS studies have tried to compare various constructs with the all-pedicle screw fixation, all have failed to address important confounding variables, such as skeletal maturity, preoperative flexibility of the curve, and factors associated with a multicenter or multisurgeon analysis., Methods: The medical records and spinal radiographs of patients with AIS treated surgically by a single surgeon between 2000 and 2009 were retrospectively reviewed. Patients with Lenke type 1 curves and minimum follow-up of 2 years were divided into 2 groups that were meticulously matched: group 1 consisted of patients in whom the all-pedicle screw construct was used, whereas group 2 included patients who were treated with the hybrid hook-screw system., Results: Group 1 included 34 patients and group 2 included 29 patients. At the last follow-up, thoracic curve correction averaged 70.4% for the all-pedicle screw group and 60% for the hybrid group (P = 0.19). The all-pedicle screw group showed a significantly greater increase in thoracic kyphosis than the hybrid group system (P = 0.04). Global sagittal balance showed greater improvement in the all-pedicle screw group during the immediate postoperative that was lost by the last follow-up. The all-pedicle screw system revealed less intraoperative blood loss but greater operating time than the hybrid construct. After controlling for length of follow-up, no statistical difference in any of the radiographical parameters measured was recorded., Conclusion: With the exception of global sagittal balance, the pedicle screw system provided better maintenance of its corrective parameters when followed for greater than two years., Level of Evidence: 3.

Study Design: Retrospective cross-sectional assessment of radiographs of adolescent idiopathic scoliosis., Objective: To assess accuracy, reliability, and reproducibility of the central sacral vertical line (CVSL) on scoliosis radiographs and its influence on determining clinically relevant parameters: stable vertebra (SV) and lumbar modifier (LM)., Background Summary: The central sacral vertical line is frequently used on scoliosis radiographs for surgical planning. However, no studies have assessed how accurately and reliably a physician draws CSVL in routine clinical practice., Methods: We provided 30 digital posteroanterior X-rays of adolescent idiopathic scoliosis to 5 raters (3 fellowship-trained spine surgeons and 2 fellows) at 2 different times (3-week interval) to determine SV and LM. An independent observer then assessed the accuracy of CSVL drawn by the raters. The CSVL was considered accurate when it was drawn vertical from the middle of the S1 vertebra. To avoid conscious bias, we kept raters blinded as to the actual purpose of the study. To avoid technical bias, the study was conducted with research Picture Archiving and Communication Systems (PACS) software and raters used desktop computers similar to those in clinics or operating rooms., Results: Based on absolute values, the CSVL was drawn on average 2.3 (±1.9) mm away from the center. Based on raw values, the lines were drawn on average 1.0 (±2.8) mm left of the center. We observed fair inter-rater reliability among the 5 raters, with an intra-class correlation of 0.23 (95% confidence interval, -0.33-0.59). Intra-rater reproducibility was moderate across 2 assessments, with an ICC of 0.47 (95% confidence interval, 0.27-0.62). Variation in drawing CSVL from center was not significantly associated with the selection of SV (r=-0.02; p=.78) or LM (r=-0.02; p=.78)., Conclusion: Although, physicians draw CSVL significantly away from the center of the S1 vertebra (mean, 2.3 mm), its influence on determining SV or LM is not affected in routine practice., (Copyright © 2013 Scoliosis Research Society. Published by Elsevier Inc. All rights reserved.)

The pathophysiology of paradoxical elbow flexion contractures following neonatal brachial plexus injury (NBPI) is incompletely understood. The current study tests the hypothesis that this contracture occurs by denervation-induced impairment of elbow flexor muscle growth. Unilateral forelimb paralysis was created in mice in four neonatal (5-day-old) BPI groups (C5-6 excision, C5-6 neurotomy, C5-6 neurotomy/repair, and C5-T1 global excision), one non-neonatal BPI group (28-day-old C5-6 excision), and two neonatal muscle imbalance groups (triceps tenotomy ± C5-6 excision). Four weeks post-operatively, motor function, elbow range of motion, and biceps/brachialis functional lengths were assessed. Musculocutaneous nerve (MCN) denervation and reinnervation were assessed immunohistochemically. Elbow flexion motor recovery and elbow flexion contractures varied inversely among the neonatal BPI groups. Contracture severity correlated with biceps/brachialis shortening and MCN denervation (relative axon loss), with no contractures occurring in mice with MCN reinnervation (presence of growth cones). No contractures or biceps/brachialis shortening occurred following non-neonatal BPI, regardless of denervation or reinnervation. Neonatal triceps tenotomy did not cause contractures or biceps/brachialis shortening, nor did it worsen those following neonatal C5-6 excision. Denervation-induced functional shortening of elbow flexor muscles leads to variable elbow flexion contractures depending on the degree, permanence, and timing of denervation, independent of muscle imbalance., (Copyright © 2012 Orthopaedic Research Society.)

1. Homomeric cyclic nucleotide-gated (CNG) channels composed of alpha2 subunits from bovine cone photoreceptors were heterologously expressed in the human embryonic kidney (HEK) 293 cell line. Modulation of cGMP sensitivity by protein kinase C (PKC)-mediated phosphorylation and by binding of calmodulin (CaM) was investigated in inside-out patches. 2. A peptide encompassing the putative CaM-binding site within the N-terminus of the channel protein binds Ca(2+)-CaM with high affinity, yet the ligand sensitivity of alpha2 channels is not modulated by CaM. 3. PKC-mediated phosphorylation increased the activation constant (K(1/2)) for cGMP from 19 to 56 microM and decreased the Hill coefficient (from 2.5 to 1.5). The change in ligand sensitivity involves phosphorylation of the serine residues S577 and S579 in the cGMP-binding domain. The increase in K(1/2) was completely abolished in mutant channels in which the two serine residues were replaced by alanine. 4. An antibody specific for the delta isoform of PKC strongly labels the cone outer segments. 5. Modulation of cGMP affinity of bovine alpha2 CNG channels by phosphorylation could play a role in the regulation of photoreceptor sensitivity.

Upon stimulation by odorants, Ca(2+) and Na(+) enter the cilia of olfactory sensory neurons through channels directly gated by cAMP. Cyclic nucleotide-gated channels have been found in a variety of cells and extensively investigated in the past few years. Glutamate residues at position 363 of the alpha subunit of the bovine retinal rod channel have previously been shown to constitute a cation-binding site important for blockage by external divalent cations and to control single-channel properties. It has therefore been assumed, but not proven, that glutamate residues at the corresponding position of the other cyclic nucleotide-gated channels play a similar role. We studied the corresponding glutamate (E340) of the alpha subunit of the bovine olfactory channel to determine its role in channel gating and in permeation and blockage by Ca(2+) and Mg(2+). E340 was mutated into either an aspartate, glycine, glutamine, or asparagine residue and properties of mutant channels expressed in Xenopus laevis oocytes were measured in excised patches. By single-channel recordings, we demonstrated that the open probabilities in the presence of cGMP or cAMP were decreased by the mutations, with a larger decrease observed on gating by cAMP. Moreover, we observed that the mutant E340N presented two conductance levels. We found that both external Ca(2+) and Mg(2+) powerfully blocked the current in wild-type and E340D mutants, whereas their blockage efficacy was drastically reduced when the glutamate charge was neutralized. The inward current carried by external Ca(2+) relative to Na(+) was larger in the E340G mutant compared with wild-type channels. In conclusion, we have confirmed that the residue at position E340 of the bovine olfactory CNG channel is in the pore region, controls permeation and blockage by external Ca(2+) and Mg(2+), and affects channel gating by cAMP more than by cGMP.

Gating by cGMP and voltage of the alpha subunit of the cGMP-gated channel from rod photoreceptor was examined with a patch-clamp technique. The channels were expressed in Xenopus oocytes. At low [cGMP] (<20 microM), the current displayed strong outward rectification. At low and high (700 microM) [cGMP], the channel activity was dominated by only one conductance level. Therefore, the outward rectification at low [cGMP] results solely from an increase in the open probability, P(o). Kinetic analysis of single-channel openings revealed two exponential distributions. At low [cGMP], the larger P(o) at positive voltages with respect to negative voltages is caused by an increased frequency of openings in both components of the open-time distribution. In macroscopic currents, depolarizing voltage steps, starting from -100 mV, generated a time-dependent current that increased with the step size (activation). At low [cGMP] (20 microM), the degree of activation was large and the time course was slow, whereas at saturating [cGMP] (7 mM) the respective changes were small and fast. The dose-response relation at -100 mV was shifted to the right and saturated at significantly lower P(o) values with respect to that at +100 mV (0.77 vs. 0.96). P(o) was determined as function of the [cGMP] (at +100 and -100 mV) and voltage (at 20, 70, and 700 microM, and 7 mM cGMP). Both relations could be fitted with an allosteric state model consisting of four independent cGMP-binding reactions and one voltage-dependent allosteric opening reaction. At saturating [cGMP] (7 mM), the activation time course was monoexponential, which allowed us to determine the individual rate constants for the allosteric reaction. For the rapid rate constants of cGMP binding and unbinding, lower limits are determined. It is concluded that an allosteric model consisting of four independent cGMP-binding reactions and one voltage-dependent allosteric reaction, describes the cGMP- and voltage-dependent gating of cGMP-gated channels adequately.

Cyclic nucleotide-gated (CNG) channels play an important role in Ca2+ signaling in many cells. CNG channels from various tissues differ profoundly in their Ca2+ permeation properties. Using the voltage-dependent Ca2+ blockage of monovalent current in wild-type channels, chimeric constructs and point mutants, we have identified structural elements that determine the distinctively different interaction of Ca2+ with CNG channels from rod and cone photoreceptors and olfactory neurons. Segments S5 and S6 and the extracellular linkers flanking the pore region are the only structural elements that account for the differences between channels. Ca2+ blockage is strongly modulated by external pH. The different pH dependence of blockage suggests that the pKa of intrapore glutamates and their protonation pattern differ among channels. The results support the hypothesis that the S5-pore-S6 module, by providing a characteristic electrostatic environment, determines the protonation state of pore glutamates and thereby controls Ca2+ affinity and permeation in each channel type.

The complete amino-acid sequence of the bovine olfactory epithelium adenosine 3',5'-cyclic monophosphate (cAMP)-gated channel has been determined by cloning and sequencing its cDNA. It exhibits a high degree of sequence homology with the cGMP-gated channel of rod photoreceptors, suggesting that cyclic nucleotide-gated channels fall into a new family of genetically related proteins.

We have analyzed protein-DNA complexes formed between lac repressor and linear or differently supercoiled lac DNA (802 or 816 base-pairs in length), which carry all three natural lac operators (O1, O2 and O3) in their wild-type sequence context and spacing and compared them with constructs that contain specifically mutated "pseudo-operators" O2 or O3. We used gel retardation assays to identify the nature of the complexes according to their characteristic electrophoretic mobility and dissociation rate measurements to determine their stability. With linear DNA we found only indirect evidence for loop formation between O1 and O2. In covalently closed DNA minicircles the formation of a loop between O1 and O2 could be demonstrated by the observation that O1-O2 containing DNA with low negative supercoiling (sigma = -0.013 and less) is constricted by binding of lac repressor, resulting in an increased electrophoretic mobility. At elevated negative supercoiling (sigma = -0.025, -0.037, -0.05) O1-O2 containing DNA complexed with lac repressor migrates significantly slower than the corresponding O1-DNA, indicating loop formation. The dissociation of lac repressor-operator complexes is decreased with increasing negative supercoiling for all tested operator combinations of O1, O2 and O3. However, in the presence of at least two natural lac operators on the same DNA minicircle the enhancement of stability is particularly large. This indicates that a DNA loop is formed between these two lac operators, O1 and O2 as well as O1 and O3, since negative supercoiling is known specifically to promote the formation of looped structures. Additionally, we observe a dependence of dissociation rate on the spatial alignment of the operators as a result of changing helical periodicity in differently supercoiled DNA and consider this to be further evidence for loop formation between O1 and O2 as well as O1 and O3.

We tested the effect of systematic destruction of all three lac operators of the chromosomal lac operon of Escherichia coli on repression by Lac repressor. Absence of just one 'pseudo-operator' O2 or O3 decreases repression by wild-type tetrameric Lac repressor approximately 2- to 3-fold; absence of both 'pseudo-operators' decreases repression greater than 50-fold. O1 alone represses under these conditions only approximately 20-fold. Dimeric active Lac repressor (iadi) represses the wild-type lac operon to about the same low extent. This indicates that cooperative interaction between lac operators is due to DNA loop formation mediated by tetrameric Lac repressor. Under conditions where loop formation is impossible, occupation of O3 but not of O2 may lead to weak repression. This suggests that under these conditions CAP activation may be inhibited and that stopping transcription at O2 does not significantly contribute to repression.

The second operator of the lac operon, located within the 5'-coding region of the lacZ gene, was specifically destroyed by means of oligonucleotide-directed mutagenesis. Eight of its bases were exchanged without altering the wild-type amino acid sequence of beta-galactosidase. The mutation was transferred onto an F'lac+I+O+Z+pro+ episome. We observed a fivefold decrease in repression of beta-galactosidase expression compared to that in the wild-type.