Dynamic radiostereometry can objectively quantify the kinematic laxity patterns and rotation instability of the knee during a pivot-shift test.

PURPOSE: The pivot-shift test is used to clinically assess knee instability in patients with anterior cruciate ligament (ACL) lesions; however, it has low interobserver reliability. Dynamic radiostereometry (dRSA) is a highly precise and noninvasive method for the objective evaluation of joint kinematics. The purpose of this study was to quantify precise knee kinematics during a pivot-shift test using dRSA imaging. METHOD: Eight human donor legs, including hemipelvises, were evaluated. Arthroscopic intervention was performed inducing ligament lesions in the ACL, and anterolateral ligament (ALL) section was performed as a capsular incision. The pivot-shift test was recorded with dRSA on knees with intact ligaments, ACL-deficient and ACL + ALL-deficient knees. RESULTS: A pivot-shift pattern was identifiable after ligament lesion, as a change in tibial posterior drawer velocity from 7.8 mm/s (95% CI: 3.7; 11.9) in ligament intact knees to 30.4 mm/s (95% CI 23.0; 38.8) after ACL lesion to 35.1 mm/s (95% CI 23.4; 46.7) after combined ACL-ALL lesion. The anterior-posterior drawer excursion increased from 2.8 mm (95% CI 2.1; 3.4) in ligament intact knees to 7.2 mm (95% CI 5.5; 8.9) after ACL lesion to 7.6 mm (95% CI 5.5; 9.8) after combined lesion. A statistically significant increase in tibial external rotation towards the end of the pivot-shift motion was observed when progressing from intact to ACL + ALL-deficient knees (p < 0.023). CONCLUSION: This experimental study demonstrates the feasibility of dRSA to objectively quantify the kinematic laxity patterns of the knee during the pivot-shift test. The dynamic parameters obtained through dRSA revealed the kinematic changes from ACL to combined ACL-ALL ligament lesion. LEVEL OF EVIDENCE: Not applicable.

Clean PDI-1 SQ: Suppression of HSQC artifacts in 2D proton-detected INADEQUATE spectra by pulse sequence redesign.

Proton-detected INADEQUATE NMR experiments are widely used for structure elucidation of small molecules, in particular the implementations that display 13C single-quantum rather than double-quantum frequencies in the indirect dimension of 2D spectra. But unfortunately, such spectra in addition to the desired 1H-13C two-bond correlations also contain HSQC artifacts of comparable magnitude. The redesigned versatile experiment presented in this paper requires no compromise based on different 13C multiplicities and suppresses the HSQC artifacts that are a source of possible spectral misinterpretation. Demonstration of the new method is shown by applications to typical small molecules of different complexity.

Risks, solutions and implementation of a participatory ergonomic intervention in 16 day nurseries: A process evaluation of the TOY wait-list cluster-randomized controlled trial.

More evidence-based initiatives to reduce physical work demands during childcare work to prevent ill health and promote the ability to care for the children among childcare workers are needed. In a process evaluation performed alongside a two-arm, cluster-randomized study with a waiting-list control among 16 day nurseries lasting 20-weeks that significantly reduced musculoskeletal pain-related sickness absence we investigated 1) risk factors and solutions perceived by the childcare workers, and 2) implementation of the intervention. Most of the perceived risk factors were categorized as physical (70 %) with most of the suggested solutions also being categorized as physical (61 %). The remaining risk factors were categorized as organizational risk factors (16 %) and psychosocial risk factors (13 %). The remaining solutions were distributed almost equally between the organizational (20 %) and psychosocial categories (19 %). About half (51 %) of the action plans showed high implementation success. Of 16 workshops, 100 % were delivered with a fidelity of 83 %. Average participation, exposure, responsiveness and implementation were 68 %, 56 %, 83 % and 47 %. The implementation score differed for timing of intervention but not for nursery characteristics. This study showed that complex and diverse participatory ergonomic interventions should focus on physical, organizational and psychosocial factors to have a positive effect.

Single- or two-stage revision after failed ACL reconstruction: No differences in re-revision rates and clinical outcomes.

PURPOSE: The surgeons' choice of a single-stage or a two-stage procedure in revision anterior cruciate ligament reconstruction (ACLr) is based on the possibility of reuse of the tibia and femoral bone tunnels after primary ACLr. The purpose of this study was to compare failure rates and clinical outcomes following single-stage and two-stage ACL revisions in a cohort of patients from The Danish Knee Ligament Reconstruction Registry. METHODS: Patients identified from 2005 to 2022 with ACL revision and met the following criteria: minimum 2-year follow-up, isolated ACL revision and registered single- or two-stage ACL revision. The primary outcome was ACL re-revision rate. Secondary outcomes were arthrometer sagittal knee laxity (side-to-side difference) and pivot shift (rotational stability difference) evaluated at 1-year follow up. RESULTS: One thousand five hundred seventy-four ACL revisions were included in the study (1331 = single-stage and 243 = two stage). Baseline characteristics showed no difference in relation to age, gender, knee laxity, pivot shift, meniscus injury, cartilage damage or injury mechanism between the two groups. Significant differences were found in relation to the type of graft. No statistical difference in 2-years revision rates between single-stage group 2.79 (95% CI 2.03-3.84) and two-stage group 2.93 (95% CI 1.41-6.05) was found. No significant difference was seen in knee laxity and pivot shift between stage-groups at 1-year follow up. Both groups demonstrated significant improvements in knee stability from baseline to 1-year follow-up. CONCLUSION: The present study found that ACL revision outcomes were similar in terms of rerevision rates and knee laxity for patients managed with a single- or a two-stage surgical strategy. LEVEL OF EVIDENCE: Level III.

Bioinformatics and functional selection of GH77 4-α-glucanotransferases for potato starch modification.

4-α-glucanotransferases (4αGTs, EC 2.4.1.25) from glycoside hydrolase family 77 (GH77) catalyze chain elongation of starch amylopectin chains and can be utilized to structurally modify starch to tailor its gelation properties. The potential relationship between the structural design of 4αGTs and functional starch modification is unknown. Here, family GH77 was mined in silico for enzyme candidates based on sub-grouping guided by Conserved Unique Peptide Patterns (CUPP) bioinformatics categorization. From + 12,000 protein sequences a representative set of 27 4αGTs, representing four different domain architectures, different bacterial origins and diverse CUPP groups, was selected for heterologous expression and further study. Most of the enzymes catalyzed starch modification, but their efficacies varied substantially. Five of the 4αGTs were characterized in detail, and their action was compared to that of the industrial benchmark enzyme, Tt4αGT (CUPP 77_1.2), from Thermus thermophilus. Reaction optima of the five 4αGTs ranged from ∼40-60 °C and pH 7.3-9.0. Several were stable for a minimum 4 h at 70 °C. Domain architecture type A proteins, consisting only of a catalytic domain, had high thermal stability and high starch modification ability. All five novel 4αGTs (and Tt4αGT) induced enhanced gelling of potato starch. One, At4αGT from Azospirillum thermophilum (CUPP 77_2.4), displayed distinct starch modifying abilities, whereas T24αGT from Thermus sp. 2.9 (CUPP 77_1.2) modified the starch similarly to Tt4αGT, but slightly more effectively. T24αGT and At4αGT are thus interesting candidates for industrial starch modification. A model is proposed to explain the link between the 4αGT induced molecular modifications and macroscopic starch gelation.

A woman in her fifties with cirrhosis of the liver and postural dyspnoea. / En kvinne i 50-årene med levercirrhose og stillingsavhengig dyspné.

A woman in her fifties with advanced cirrhosis of the liver was admitted multiple times with recurrent pleural effusion and ascites. She was accepted for liver transplantation, at which time she developed postural dyspnoea and a drop in oxygen saturation.

Combined anterior cruciate ligament revision with reconstruction of the antero-lateral ligament does not improve outcome at 2-year follow-up compared to isolated acl revision; a randomized controlled trial.

PURPOSE: It is essential to obtain rotational stability of the knee after anterior cruciate ligament reconstruction (ACL-R) and it is suggested that a supplementary reconstruction of the antero-lateral ligament (ALL-R) may supports this. Theoretically, ALL-R may be particularly advantageous to support revision of failed ACL-Rs. It was hypothesized that ACL revision combined with ALL-R will result in superior outcome compared to isolated ACL revision. METHODS: The study was designed as a randomized controlled trial. Patients eligible for first time ACL revision were randomized to either isolated ACL revision (- ALL group) or ACL revision combined with a single-stranded allograft ALL-reconstruction (+ ALL group). Patient reported outcomes and function were evaluated at two-year follow-up by KNEES-ACL, KOOS, and Tegner activity scale. Objective knee laxity was evaluated at one-year follow-up using an instrumented Rolimeter test, the pivot shift test, and a manual Lachman test. RESULTS: A total of 103 patients were enrolled with 49 patients randomized to the + ALL group and 54 patients in the - ALL group. There were no differences at baseline between groups regarding age, gender, body mass index, preoperative patient reported outcome scores and concomitant meniscus or cartilage injury. The ACL revision was performed with an allograft in 10 patients (20%) in the + ALL group and 8 patients (15%) in the -ALL group. At follow-up there was no significant difference between the groups in patient reported outcome scores and clinical knee laxity. CONCLUSION: Supplementary ALL-R does not improve subjective outcome of first time ACL revision at two-years and clinical knee stability at one-year follow-up compared to isolated ACL revision. LEVEL OF EVIDENCE: Level I.

Origin and remedy for HSQC artifacts in proton-detected INADEQUATE spectra.

NMR pulse sequences visualizing 1JCC and nJCC bond connectivity via an intermediate state of 13C-13C double-quantum coherence and 1H detection are an indispensable tool to solve small-molecule structures at the natural abundance level of 13C. A longstanding issue with these experiments set up to display 2D spectra with single-quantum frequencies is that in addition to the 1H-13C-13C correlations of interest, appearance of HSQC-type artifacts can complicate analysis and obscure JCC connectivities. The origin of these artifacts is described and remedies for their suppression are introduced. They include refocusing of 1JCH couplings prior to creation of 13C-13C double-quantum coherence, which is known to enhance sensitivity by reducing loss into zero-quantum coherence for pairs of two protonated 13C.

Comparing spatial management tools to protect highly migratory shark species in the Eastern Mediterranean Sea hot spots.

Bycatch of non-target species is a pressing problem for ocean management. It is one of the most concerning issues related to human-wildlife interactions and it affects numerous species including sharks, seabirds, sea turtles, and many critically endangered marine mammals. This paper compares different policy tools for ocean closure management around a unique shark aggregation site in Israel's nearshore coastal waters. We provide a set of recommendations based on an optimal management approach that allows humans to enjoy marine recreational activities such as fishing, while maintaining safe conditions for these apex predators which are vital to the local marine ecosystem. To learn more about recreational fishers' derived benefits, we use a benefit transfer method. Our main conclusion is that dynamic time-area closures offer sustainable and effective management strategies. Since these closures are based on near real-time data, they might successfully preserve specific species in limited areas (i.e., small areas).

Importance of Inactivation Methodology in Enzymatic Processing of Raw Potato Starch: NaOCl as Efficient α-Amylase Inactivation Agent.

Efficient inactivation of microbial α-amylases (EC 3.2.1.1) can be a challenge in starch systems as the presence of starch has been shown to enhance the stability of the enzymes. In this study, commonly used inactivation methods, including multistep washing and pH adjustment, were assessed for their efficiency in inactivating different α-amylases in presence of raw potato starch. Furthermore, an effective approach for irreversible α-amylase inactivation using sodium hypochlorite (NaOCl) is demonstrated. Regarding inactivation by extreme pH, the activity of five different α-amylases was either eliminated or significantly reduced at pH 1.5 and 12. However, treatment at extreme pH for 5 min, followed by incubation at pH 6.5, resulted in hydrolysis yields of 42-816% relative to controls that had not been subjected to extreme pH. "Inactivation" by multistep washing with water, ethanol, and acetone followed by gelatinization as preparation for analysis gave significant starch hydrolysis compared to samples inactivated with NaOCl before the wash. This indicates that the further starch degradation observed in samples subjected to washing only took place during the subsequent gelatinization. The current study demonstrates the importance of inactivation methodology in α-amylase-mediated raw starch depolymerization and provides a method for efficient α-amylase inactivation in starch systems.

Effects of topical corticosteroid versus tacrolimus on insulin sensitivity and bone homeostasis in adults with atopic dermatitis-A randomized controlled study.

INTRODUCTION: Topical corticosteroids (TCS), used to treat atopic dermatitis (AD), have been associated with type 2 diabetes and osteoporosis in epidemiological studies, possibly explained by systemic absorption. OBJECTIVES: We examined whether intensive daily whole-body TCS treatment over 2 weeks followed by twice weekly application for 4 weeks could elicit insulin resistance and increase bone resorption in adults with AD. METHODS: A randomized parallel-group double-blind double-dummy non-corticosteroid-based active comparator study design was completed in Copenhagen, Denmark. Thirty-six non-obese, non-diabetic adults with moderate-to-severe AD were randomized to whole-body treatment with betamethasone 17-valerate 0.1% plus a vehicle once daily or tacrolimus 0.1% twice daily after washout. Insulin sensitivity assessed by the hyperinsulinemic-euglycemic clamp combined with tracer infusions and biomarkers of bone formation (P1NP) and resorption (CTX) were evaluated at baseline, after 2 weeks of daily treatment and after further 4 weeks of twice-weekly maintenance treatment. RESULTS: AD severity improved with both treatments and systemic inflammation was reduced. After 2 weeks, we observed similar increase in peripheral insulin sensitivity with use of betamethasone (n = 18) and tacrolimus (n = 18). Bone resorption biomarker, CTX, was unchanged, while bone formation marker, P1NP, decreased after betamethasone treatment after both 2 and 6 weeks but remained unchanged in the tacrolimus arm. CONCLUSIONS: Whole-body treatment with TCS leads to systemic exposure but appears not to compromise glucose metabolism during short-term use, which may be a result of reduced systemic inflammatory activity. The negative impact on bone formation could be regarded an adverse effect of TCS.

Enzymatic potato starch modification and structure-function analysis of six diverse GH77 4-alpha-glucanotransferases.

4-α-glucanotransferase (EC 2.4.1.25) mediated glucan transfer in starch provides opportunities for production of clean label starch ingredients with unique gelling properties. 4-α-glucanotransferases can be found in glycoside hydrolase (GH) family GH13, GH57, and in the monospecific glycoside hydrolase family 77 (GH77). Here, pH-temperature optima, steady-state kinetics, potato starch modifying properties and structural folds are reported for six phylogenetically distinct GH77 members, representing four different domain architectures including a novel multi-domain 4-α-glucanotransferase from Lactococcus lactis. Four of the enzymes exhibited starch modifying activity leading to a gradual decrease of the amylose content, elongation of amylopectin chains, and enabled formation of firm starch gels. Unexpectedly, these diverse enzymes catalyzed similar changes in chain length distributions. However, the amylose depletion and amylopectin elongation rates spanned more than two orders of magnitude between the enzyme showing very different specific activities. Tt4αGT from Thermus thermophilus had highest temperature optimum (73 °C) and superior potato starch modifying efficacy compared to the other five enzymes.

Four-in-one: HSQC, HSQC-TOCSY (or H2BC), TOCSY, and enhanced HMBC spectra integrated into a single NO Relaxation Delay (NORD) NMR experiment.

The NMR pulse sequence design strategy of NORD (NO Relaxation Delay) is extended to design of two new three-module experiments, NORD {HMBC}-{HSQC-TOCSY}-{TOCSY} and NORD {HMBC}-{2BOB}-{TOCSY}, each delivering four spectra - HMBC, HSQC, TOCSY, and either HSQC-TOCSY or H2BC. Compared to individual recording of these spectra particularly the sensitivity of the least sensitive module, HMBC, is enhanced by designing the homonuclear TOCSY module to allow buildup of magnetization pertinent to HMBC during its execution. Effectively, the sensitivity of the heteronuclear modules is boosted at the expense of the inherently much higher TOCSY sensitivity, thus resulting in a significant saving in spectrometer time.

A comparison of multi-ligament reconstruction and isolated anterior cruciate ligament reconstruction at one year follow-up: results from the Danish Knee Ligament Reconstruction Registry.

INTRODUCTION: The Danish Knee Ligament Reconstruction Registry (DKRR) has monitored the outcomes of surgeries for multi-ligament knee injuries (MLKI) since 2005. This study aimed to compare the subjective clinical outcomes of patients who had undergone surgery after MLKI with those of patients who had received isolated anterior cruciate ligament (ACL) reconstruction. MATERIALS AND METHODS: This study used patient-reported outcome scores at 1-year follow-up as the primary outcome and contains the outcome data of knee ligament surgeries retrieved from the DKRR. Clinical subjective outcomes and knee function were evaluated with Knee Injury and Osteoarthritis Outcome Scores (KOOS) and Tegner Activity Scale (Tegner) scores. Demographic differences were examined using the Student's t-test and the chi-square test. Multiple linear regression was used to analyse the data and adjust for potentially confounding factors. P-values < 0.05 were considered to be statistically significant. RESULTS: A total of 31,686 knee ligament surgeries were registered in the DKRR between 2005 and 2017, resulting in 1,160 multi-ligament patients and 28,843 isolated ACL patients. The mean age of the MLKI group was significantly higher than that of the isolated ACL group (33.2 years [95% CI 32.5-33.9] vs. 28.3 years [95% CI 28.1-28.4]). The adjusted KOOS Sport and Quality of Life (QoL) sub-scores and Tegner scores of the MLKI group significantly improved from the baseline to the 1-year follow-up (16.7 points [95%CI 12.8;20.6], 12.6 points [95%CI 9.6;15.6] and 1.76 points [95%CI 1.43;2.08], respectively). The KOOS Sport and QoL sub-scores of the isolated ACL group were significant and increased more than those of the MLKI group. No differences in the Tegner scores were observed. CONCLUSIONS: Surgical reconstruction after multi-ligament knee injury resulted in significant subjective outcome improvements at 1- year follow-up. The KOOS Sport and QoL sub-scores of the isolated ACL group significantly increased compared to those of the MLKI group.

Synergy and sensitivity-balance in concatenating experiments in NO relaxation delay NMR (NORD).

The NMR experiment design strategy of NO Relaxation Delay (NORD), introduced mostly as an idealized theoretical approach, is extended and put to practical use by considering synergy and sensitivity-balance in concatenation of experiments. It is illustrated by a novel experiment, NORD {HMBC}-{HSQC}-{TOCSY}, where magnetization of non-13C attached protons effectively is channeled from the TOCSY spectrum toward primarily the least sensitive spectrum of HMBC. The experiment is expected to find its place as a full-package NMR method for metabolomics, carbohydrates, peptides and small-molecules in general.

NORD: NO Relaxation Delay NMR Spectroscopy.

The novel concept of NORD (NO relaxation delay) NMR spectroscopy is introduced. The idea is to design concatenated experiments in a way that the magnetization used in the first relaxes toward equilibrium during the second and vice versa, thus saving instrument time. Applications include complete well-resolved 1 H-1 H and 1 H-13 C one-bond and long-range correlation maps of an 80 mM solution of a trisaccharide recorded in less than two minutes and hydrocortisone with extensive spectral overlap.

Effects of a participatory organisational, core work task focused workplace intervention on employees' primary healthcare consultations: secondary analysis of a cluster RCT.

OBJECTIVES: We aimed to examine whether a participatory organisational workplace intervention focusing on core tasks at work resulted in lower primary healthcare utilisation of employees. METHODS: The cluster randomised controlled trial included 78 preschools, 44 allocated to the intervention group (1745 employees) and 34 allocated to the control group (1267 employees). The intervention aimed to involve employees in improving the psychosocial work environment while focusing on core tasks at work. Using Poisson regression, we tested the rate ratios (RRs) of consultations in the intervention compared with the control group in terms of all consultations in primary healthcare and general practitioner (GP) consultations, respectively, per person-year during 31 months of follow-up. The fully adjusted model included adjustment for sex, age, job group, workplace type and size, and previous primary healthcare utilisation. RESULTS: During the follow-up, intervention group employees had 11.0 consultations/person-year, while control group employees had 11.6 consultations/person-year (RR 0.97, 95% CI 0.92 to 1.01). Employees in the intervention group had 7.5 GP consultations/person-year, while control group employees had 8.2 GP consultations/person-year (RR 0.95, 95% CI 0.90 to 0.99). Post hoc analyses indicated that the effect of the intervention was particularly strong in employees in preschools with a moderate or high level of implementation. CONCLUSIONS: The participatory organisational workplace intervention focusing on core tasks at work among preschool employees had a small, statistically non-significant effect on overall primary healthcare utilisation and a small, statistically significant effect on GP consultations. These results suggest a beneficial effect of the participatory organisational intervention on employees' health. TRIAL REGISTRATION NUMBER: ISRCTN16271504.

Using chemiluminescence imaging of cells (CLIC) for relative protein quantification.

Cell physiology and cellular responses to external stimuli are partly controlled through protein binding, localization, and expression level. Thus, quantification of these processes is pivotal in understanding cellular biology and disease pathophysiology. However, it can be methodologically challenging. Immunofluorescence is a powerful technique, yet quantification by this method can be hampered by auto-fluorescence. Here we describe a simple, sensitive and robust chemiluminescence-based immunoassay (chemiluminescence imaging of cells; CLIC) for relative quantification of proteins. We first employed this method to quantify complement activation in cultured mammalian cells, and to quantify membrane protein expression, shedding, binding and internalization. Moreover, through specific membrane permeabilization we were able to quantify both cytosolic and nuclear proteins, and their translocation. We validated the CLIC quantification method by performing parallel experiments with other quantification methods like ELISA, qPCR, and immunofluorescence microscopy. The workflow of the immunoassay was found to be advantageous in certain instances when compared to these quantification methods. Since the reagents used for CLIC are common to other immunoassays with no need for specialized equipment, and due to the good linearity, dynamic range and signal stability inherent to chemiluminescence, we suggest that this assay is suitable for both small scale and high throughput relative protein quantification studies in whole cells.

Bone ingrowth into open architecture PEEK interference screw after ACL reconstruction.

PURPOSE: Open or fenestrated interference screw design that allow bone ingrowth is a concept for improved bone healing to softtissue graft and bone filling in bone tunnels after anterior cruciate ligament reconstruction (ACLR) The aim of the current study was to assess CT scanning evaluated bone ingrowth into an open architecture interference screws in the tibial tunnel of patients undergoing ACL with soft tissue grafts. It was hypothesized that open architecture interference screws would stimulate bone ingrowth into the screw cavities. METHODS: Twelve patients requiring ACLR were included. They underwent arthroscopic ACLR with semitendinosus-/gracilis tendon graft and an open architecture polyetheretherketone (PEEK) interference screw. The patients were scanned with a multi-slice CT scanner two weeks, six and twelve months postoperatively. On CT reconstruction slices bone ingrowth into the screw was measured. Subjective and objective clinical outcome international knee documentation committee score and instrumented knee laxity determination were collected. RESULTS: At six months no implants demonstrated more than 10% bone ingrowth. At twelve months 42% (5/12) implants had more than 10% bone ingrowth (p = 0.009). The average bone filling into the screws was 7.7%. There was no tunnel widening or cyst formation seen in relation to any of the implants. Subjective IKDC score improved significantly from 50.6 baseline to 80.1 at 24 month follow-up. Preoperative side-to-side knee laxity improved from 3.7 (2.1) to 1.4 (1.2) mm at twelve months. There were no serious adverse events in relation to the new open architecture thread PEEK interference screw during or after hamstring ACL reconstruction. CONCLUSION: The present study demonstrated that open architecture thread PEEK interference screw can stimulate bone ingrowth into the screws after soft tissue ACL reconstruction with at 12 months with an average bone filling into screws was 7.7%. Knee stability, functional, subjective and objective outcomes were similar to large volume ACL outcome studies. TRIAL REGISTRATION: The study was registered at ClinicalTrials # NCT02382341 . 12-09-2014. LEVEL OF EVIDENCE: IV.

Double and adiabatic BANGO for concatenating two NMR experiments relying on the same pool of magnetization.

It is shown how the same pool of magnetization can be tapped twice in two different concatenations of three experiments into a single pulse sequence with only one relaxation delay. This is accomplished by using the BANGO pulse sequence element twice for independent rotations of 1H magnetization attached or not attached to 13C and it includes a refinement of BANGO with an adiabatic 13C inversion pulse resulting in improved tolerance to a spread in 1JCH coupling constants that translates directly into improved sensitivity of the modular experiment relying on 1H magnetization attached to 13C. The two new pulse sequences are SEA XLOC-HMBC-H2OBC/2BOB and SEA XLOC(ZQ)-SEA XLOC(2Q)-H2OBC/2BOB which both represent a rapid route to complete heteronuclear one-bond and long-range JCH correlation maps for small molecules, as is demonstrated on ibuprofen and prednisolone.