Varus osteotomy as a salvage procedure for young patients with symptomatic patellofemoral arthritis and valgus malalignment at short- to mid-term follow-up: a case series.

PURPOSE: The purpose of the study was to report the clinical, functional and radiological outcome following varus osteotomy as a salvage procedure in young to middle-aged patients with patellofemoral arthritis (PFA) and associated valgus malalignment. It was hypothesized that a significant improvement in knee function and reduction in pain would be achieved. Moreover, no conversion to patellofemoral joint arthroplasty could be observed. MATERIAL AND METHODS: Patients (< 50 years of age) that underwent varus osteotomy between 08/2012 and 01/2020 for the treatment of symptomatic PFA and associated valgus malalignment were consecutively included (minimum follow-up: 24 months). Patient-reported outcome measures (PROM; International Knee Documentation Committee subjective knee form [IKDC]), Visual Analog Scale [VAS] for pain, Tegner Activity Scale [TAS], and satisfaction with the postoperative results (1-10-scale, 10 = highest satisfaction) and weight-bearing whole-leg anteroposterior radiographs were conducted pre- and postoperatively. The change in PROM and femorotibial angle (FTA) were tested for statistical significance. RESULTS: In total, 12 patients (14 knees) were included (66.7% female; mean age: 33.8 ± SD 6.6 years). In ten cases, lateral opening-wedge distal femoral osteotomies (DFO) were performed, of which three cases included a concomitant femoral derotation. Three medial closing-wedge DFO and one medial closing-wedge high tibial osteotomy were performed. At follow-up (55.3 ± 29.3 months), a significant improvement in knee function (IKDC: 56.4 ± 14.4 to 69.1 ± 11.2, p = 0.015) and reduction in pain (VAS for pain: 3.5 [interquartile range 2.3-5.8] to 0.5 [0-2.0], p = 0.018) were observed. Patients were able to reach their preoperative sporting activity level (TAS: 3.0 [3.0-4.0] to 3.5 [3.0-4.0], p = 0.854) and were highly satisfied with the postoperative result (9.0 [6.5-10]). Additionally, a significant correction of valgus malalignment was observed (5.0° ± 2.9° valgus to 0.7° ± 3.2° varus, p < 0.001). Regarding complications, two re-osteosyntheses were performed due to loss of correction and delayed union. No conversion to patellofemoral arthroplasty occurred. CONCLUSION: In patients with symptomatic PFA and associated valgus malalignment, varus osteotomy as a salvage procedure achieved a significant improvement in knee function and reduction in pain. No conversion to patellofemoral joint arthroplasty occurred at short- to mid-term follow-up. LEVEL OF EVIDENCE: Retrospective case series, Level IV.

Saccharomyces boulardii promoters for control of gene expression in vivo.

BACKGROUND: Interest in the use of engineered microbes to deliver therapeutic activities has increased in recent years. The probiotic yeast Saccharomyces boulardii has been investigated for production of therapeutics in the gastrointestinal tract. Well-characterised promoters are a prerequisite for robust therapeutic expression in the gut; however, S. boulardii promoters have not yet been thoroughly characterised in vitro and in vivo. RESULTS: We present a thorough characterisation of the expression activities of 12 S. boulardii promoters in vitro in glucose, fructose, sucrose, inulin and acetate, under both aerobic and anaerobic conditions, as well as in the murine gastrointestinal tract. Green fluorescent protein was used to report on promoter activity. Promoter expression was found to be carbon-source dependent, with inulin emerging as a favourable carbon source. Furthermore, relative promoter expression in vivo was highly correlated with expression in sucrose (R = 0.99). CONCLUSIONS: These findings provide insights into S. boulardii promoter activity and aid in promoter selection in future studies utilising S. boulardii to produce therapeutics in the gut.

[Treatment of osteochondritis dissecans]. / Therapie der Osteochondrosis dissecans.

Osteochondritis dissecans (OD) is a rare condition with an incidence of 30/100,000. It especially affects male patients aged 10-20 years old. During the staged progression the osteochondral fragments can detach from their base. These can damage the adjacent articular cartilage, which can lead to premature osteoarthritis. Most commonly affected are the knee, ankle and elbow joints. The exact pathogenesis of OD has so far not been clearly confirmed. Several risk factors that can lead to the development of OD are discussed. These include repeated microtrauma and vascularization disorders that can lead to ischemia of the subchondral bone and to a separation of the fragments close to the joint and therefore to the development of free joint bodies. For an adequate clarification patients should undergo a thorough radiological evaluation including X­ray imaging followed by magnetic resonance imaging (MRI) to assess the integrity of the cartilage-bone formation with determination of the OD stage. The assessment is based on criteria of the International Cartilage Repair Society (ICRS). The instability of the cartilage-bone fragment increases with higher stages. Stages I and II with stable cartilage-bone interconnection can be treated conservatively. For stages III and IV, i.e., instability of the OD fragment or the presence of free fragments, surgical treatment should be performed. Primarily, refixation of a free joint body should be carried out depending on the size and vitality of the fragment. In cases of unsuccessful conservative treatment or fixation, a debridement, if necessary in combination with a bone marrow stimulating procedure, can be employed corresponding to the size of the defect. For larger cartilage defects, an osteochondral graft transplantation should be considered. Overall, OD lesions in stages I and II show a good healing tendency under conservative treatment. In cases of incipient unstable OD, refixation can also lead to good clinical and radiological results.

Engineered phage with antibacterial CRISPR-Cas selectively reduce E. coli burden in mice.

Antibiotic treatments have detrimental effects on the microbiome and lead to antibiotic resistance. To develop a phage therapy against a diverse range of clinically relevant Escherichia coli, we screened a library of 162 wild-type (WT) phages, identifying eight phages with broad coverage of E. coli, complementary binding to bacterial surface receptors, and the capability to stably carry inserted cargo. Selected phages were engineered with tail fibers and CRISPR-Cas machinery to specifically target E. coli. We show that engineered phages target bacteria in biofilms, reduce the emergence of phage-tolerant E. coli and out-compete their ancestral WT phages in coculture experiments. A combination of the four most complementary bacteriophages, called SNIPR001, is well tolerated in both mouse models and minipigs and reduces E. coli load in the mouse gut better than its constituent components separately. SNIPR001 is in clinical development to selectively kill E. coli, which may cause fatal infections in hematological cancer patients.

Changes in liver metabolic pathways demonstrate efficacy of the combined dietary and microbial therapeutic intervention in MASLD mouse model.

OBJECTIVE: Metabolic dysfunction-associated steatotic liver disease (MASLD), formerly known as non-alcoholic fatty liver disease (NAFLD), is the most prevalent liver disease globally, yet no therapies are approved. The effects of Escherichia coli Nissle 1917 expressing aldafermin, an engineered analog of the intestinal hormone FGF19, in combination with dietary change were investigated as a potential treatment for MASLD. METHODS: MASLD was induced in C57BL/6J male mice by American lifestyle-induced obesity syndrome diet and then switched to a standard chow diet for seven weeks. In addition to the dietary change, the intervention group received genetically engineered E. coli Nissle expressing aldafermin, while control groups received either E. coli Nissle vehicle or no treatment. MASLD-related plasma biomarkers were measured using an automated clinical chemistry analyzer. The liver steatosis was assessed by histology and bioimaging analysis using Fiji (ImageJ) software. The effects of the intervention in the liver were also evaluated by RNA sequencing and liquid-chromatography-based non-targeted metabolomics analysis. Pathway enrichment studies were conducted by integrating the differentially expressed genes from the transcriptomics findings with the metabolites from the metabolomics results using Ingenuity pathway analysis. RESULTS: After the intervention, E. coli Nissle expressing aldafermin along with dietary changes reduced body weight, liver steatosis, plasma aspartate aminotransferase, and plasma cholesterol levels compared to the two control groups. The integration of transcriptomics with non-targeted metabolomics analysis revealed the downregulation of amino acid metabolism and related receptor signaling pathways potentially implicated in the reduction of hepatic steatosis and insulin resistance. Moreover, the downregulation of pathways linked to lipid metabolism and changes in amino acid-related pathways suggested an overall reduction of oxidative stress in the liver. CONCLUSIONS: These data support the potential for using engineered microbial therapeutics in combination with dietary changes for managing MASLD.

Cold Exposure and Oral Delivery of GLP-1R Agonists by an Engineered Probiotic Yeast Strain Have Antiobesity Effects in Mice.

Advanced microbiome therapeutics (AMTs) holds promise in utilizing engineered microbes such as bacteria or yeasts for innovative therapeutic applications, including the in situ delivery of therapeutic peptides. Glucagon-like peptide-1 receptor agonists, such as Exendin-4, have emerged as potential treatments for type 2 diabetes and obesity. However, current administration methods face challenges with patient adherence and low oral bioavailability. To address these limitations, researchers are exploring improved oral delivery methods for Exendin-4, including utilizing AMTs. This study engineered the probiotic yeast Saccharomyces boulardii to produce Exendin-4 (Sb-Exe4) in the gastrointestinal tract of male C57BL/6 mice to combat diet-induced obesity. The biological efficiency of Exendin-4 secreted by S. boulardii was analyzed ex vivo on isolated pancreatic islets, demonstrating induced insulin secretion. The in vivo characterization of Sb-Exe4 revealed that when combined with cold exposure (8 °C), the Sb-Exe4 yeast strain successfully suppressed appetite by 25% and promoted a 4-fold higher weight loss. This proof of concept highlights the potential of AMTs to genetically modify S. boulardii for delivering active therapeutic peptides in a precise and targeted manner. Although challenges in efficacy and regulatory approval persist, AMTs may provide a transformative platform for personalized medicine. Further research in AMTs, particularly focusing on probiotic yeasts such as S. boulardii, holds great potential for novel therapeutic possibilities and enhancing treatment outcomes in diverse metabolic disorders.

Physiology of Saccharomyces cerevisiae during growth on industrial sugar cane molasses can be reproduced in a tailor-made defined synthetic medium.

Fully defined laboratory media have the advantage of allowing for reproducibility and comparability of results among different laboratories, as well as being suitable for the investigation of how different individual components affect microbial or process performance. We developed a fully defined medium that mimics sugarcane molasses, a frequently used medium in different industrial processes where yeast is cultivated. The medium, named 2SMol, builds upon a previously published semi-defined formulation and is conveniently prepared from some stock solutions: C-source, organic N, inorganic N, organic acids, trace elements, vitamins, Mg + K, and Ca. We validated the 2SMol recipe in a scaled-down sugarcane biorefinery model, comparing the physiology of Saccharomyces cerevisiae in different actual molasses-based media. We demonstrate the flexibility of the medium by investigating the effect of nitrogen availability on the ethanol yield during fermentation. Here we present in detail the development of a fully defined synthetic molasses medium and the physiology of yeast strains in this medium compared to industrial molasses. This tailor-made medium was able to satisfactorily reproduce the physiology of S. cerevisiae in industrial molasses. Thus, we hope the 2SMol formulation will be valuable to researchers both in academia and industry to obtain new insights and developments in industrial yeast biotechnology.

Dual Bracing for Ulnar Collateral Ligament Injuries Restores Native Valgus Laxity and Native Medial Joint Gapping of the Elbow.

Background: Despite growing evidence on the role of the posterior ulnar collateral ligament (pUCL) in elbow stability, current ligament bracing techniques are mainly focused on the anterior ulnar collateral ligament (aUCL). A dual-bracing technique combines the repair of the pUCL and aUCL with a suture augmentation of both bundles. Purpose: To biomechanically assess a dual-bracing approach addressing aUCL and pUCL for humeral-sided complete UCL lesions to restore medial elbow laxity without overconstraining. Study Design: Controlled laboratory study. Methods: A total of 21 unpaired human elbows (11 right, 10 left; 57.19 ± 11.7 years) were randomized into 3 groups to compare dual bracing with aUCL suture augmentation and aUCL graft reconstruction. Laxity testing was performed with 25 N applied 12 cm distal to the elbow joint for 30 seconds at randomized flexion angles (0°, 30°, 60°, 90°, and 120°) for the native condition and then for each surgical technique. A calibrated motion capture system was used for assessment, allowing the 3-dimensional displacement during the complete valgus stress cycle between the optical trackers to be quantified as joint gap and laxity. The repaired constructs were then cyclically tested through a materials testing machine starting with 20 N for 200 cycles at a rate of 0.5 Hz. The load was increased stepwise by 10 N for 200 cycles until displacement reached 5.0 mm or complete failure occurred. Results: Dual bracing and aUCL bracing resulted in significantly (P = .045) less joint gapping at 120° of flexion compared with aUCL reconstruction. No significant differences in valgus laxity were found among the surgical techniques. Within each technique, there were no significant differences between the native and the postoperative state in valgus laxity and joint gapping. No significant differences between the techniques were observed in cycles to failure and failure load. Conclusion: Dual bracing restored native valgus joint laxity and medial joint gapping without overconstraining and provided similar primary stability regarding failure outcomes as established techniques. Furthermore, it was able to restore joint gapping in 120° of flexion significantly better than aUCL reconstruction. Clinical Relevance: This study provides biomechanical data on the dual-bracing approach that may help surgeons to consider this new method of addressing acute humeral UCL lesions.

Biocontainment strategies for in vivo applications of Saccharomyces boulardii.

The human gastrointestinal tract is a complex and dynamic environment, playing a crucial role in human health. Microorganisms engineered to express a therapeutic activity have emerged as a novel modality to manage numerous diseases. Such advanced microbiome therapeutics (AMTs) must be contained within the treated individual. Hence safe and robust biocontainment strategies are required to prevent the proliferation of microbes outside the treated individual. Here we present the first biocontainment strategy for a probiotic yeast, demonstrating a multi-layered strategy combining an auxotrophic and environmental-sensitive strategy. We knocked out the genes THI6 and BTS1, causing thiamine auxotrophy and increased sensitivity to cold, respectively. The biocontained Saccharomyces boulardii showed restricted growth in the absence of thiamine above 1 ng/ml and exhibited a severe growth defect at temperatures below 20°C. The biocontained strain was well tolerated and viable in mice and demonstrated equal efficiency in peptide production as the ancestral non-biocontained strain. In combination, the data support that thi6∆ and bts1∆ enable biocontainment of S. boulardii, which could be a relevant chassis for future yeast-based AMTs.

Superior Capsular Reconstruction Partially Restores Native Glenohumeral Joint Loads in a Dynamic Biomechanical Shoulder Model.

PURPOSE: To evaluate the effect of an irreparable posterosuperior rotator cuff tear (PSRCT) on glenohumeral joint loads and to quantify improvement after superior capsular reconstruction (SCR) using an acellular dermal allograft. METHODS: Ten fresh-frozen cadaveric shoulders were tested using a validated dynamic shoulder simulator. A pressure mapping sensor was placed between the humeral head and glenoid surface. Each specimen underwent the following conditions: (1) native, (2) irreparable PSRCT, and (3) SCR using a 3-mm-thick acellular dermal allograft. Glenohumeral abduction angle (gAA) and superior humeral head migration (SM) were measured using 3-dimensional motion-tracking software. Cumulative deltoid force (cDF) and glenohumeral contact mechanics, including glenohumeral contact area and glenohumeral contact pressure (gCP), were assessed at rest, 15°, 30°, 45°, and maximum angle of glenohumeral abduction. RESULTS: The PSRCT resulted in a significant decrease of gAA along with an increase in SM, cDF, and gCP (P < .001, respectively). SCR did not restore native gAA (P < .001); however, SM was significantly reduced (P < .001). Further, SCR significantly reduced deltoid forces at 30° (P = .007) and 45° of abduction (P = .007) when compared with the PSRCT. SCR did not restore native cDF at 30° (P = .015), 45° (P < .001), and maximum angle (P < .001) of glenohumeral abduction. Compared with the PSRCT, SCR resulted in a significant decrease of gCP at 15° (P = .008), 30° (P = .002), and 45° (P = .006). However, SCR did not completely restore native gCP at 45° (P = .038) and maximum abduction angle (P = .014). CONCLUSIONS: In this dynamic shoulder model, SCR only partially restored native glenohumeral joint loads. However, SCR significantly decreased glenohumeral contact pressure, cumulative deltoid forces, and superior migration, while increasing abduction motion, when compared with the posterosuperior rotator cuff tear. CLINICAL RELEVANCE: These observations raise concerns regarding the true joint-preserving potential of SCR for an irreparable posterosuperior rotator cuff tear, along with its ability to delay progression of cuff tear arthropathy and eventual conversion to reverse shoulder arthroplasty.

Arthroscopic rod technique compared to stress ultrasound in the dynamic evaluation of lateral ligament instabilities of the elbow.

INTRODUCTION: The purpose was to compare the arthroscopic rod technique to stress ultrasound in the dynamic assessment of lateral elbow instabilities. MATERIALS AND METHODS: Fifteen elbows of eight specimen with a mean age of 66.4 ± 13.3 years were assessed in a prone position following a defined dissection setup. After evaluation of the native status, an arthroscopic dissection of the radial collateral ligament (RCL) or lateral ulnar collateral ligament (LUCL), and finally of entire capsuloligamentous structures was performed. Three raters examined each state (native, RCL or LUCL lesion, complete lesion) with the arthroscopic rod technique in 90° flexion and with stress ultrasound in 30 and 90° flexion. The intra-class correlation coefficient (ICC) was calculated to assess the interrater reliability as well as test-retest reliability for each testing modality (arthroscopy and ultrasound). RESULTS: The arthroscopic rod technique showed a superior interrater and test-retest reliability of 0.953 and 0.959 (P < 0.001), respectively, when compared to stress ultrasound with an ICC of 0.4 and 0.611 (P < 0.001). A joint space opening during arthroscopy of > 6 mm humero-ulnar or > 7 mm humero-radial was indicative for a lateral collateral ligament lesion. However, a differentiation between an isolated RCL or LUCL tear was not possible. A lateral joint opening of ≥ 9 mm was only observed in complete tears of the lateral capsuloligamentous complex. CONCLUSIONS: The arthroscopic rod technique showed a superior interrater and test-retest reliability when compared to stress ultrasound. Arthroscopic assessment for radial elbow instability was found to be reliable and reproducible. A joint gapping ≥ 9 mm in the arthroscopic evaluation is a sign for a complete insufficiency of the radial capsuloligamentous complex. However, it is not possible to precisely distinguish between a lesion of the RCL or LUCL by arthroscopy. On the basis of our results, dynamic ultrasound testing may be inappropriate to objectify lateral elbow instability.

Examination of corneal microstructure in the quiescent phase of vernal keratoconjunctivitis using in vivo confocal microscopy.

INTRODUCTION: This study investigated the morphological characteristics of corneal microstructure in the quiescent phase of vernal keratoconjunctivitis (VKC). METHODS: Twenty patients with quiescent VKC and 25 healthy subjects were included. In vivo confocal microscopy (IVCM) of the central cornea was performed. Cellular density of each layer and the morphology of subbasal nerve plexus (SBNP) was analysed. Langerhans cell density (LCD), morphology (LCM), and field area (LCF) were also examined. RESULTS: No differences were found either in cell densities nor in SBNP morphology (p > 0.05). LCD, LCM and LCF were significantly higher in the VKC group (p = 0.005, p < 0.001 and p < 0.001, respectively). The severity of papillary hypertrophy had a significant impact on LCD, LCM and LCF (ß-coefficient: 19.541, p < 0.001; ß-coefficient: 0.283, p < 0.001 and ß-coefficient: 595.255, p < 0.001, respectively). DISCUSSION: In quiescent VKC, LCD, LCM, and LCF were increased, and they were associated with the severity of papillary hypertrophy. Alterations of Langerhans cells indicate a subclinical inflammatory process without ocular symptoms.

Laboratory performance prediction using virtual reality behaviometrics.

In this study, we show that virtual reality (VR) behaviometrics can be used for the assessment of compliance and physical laboratory skills. Drawing on approaches from machine learning and classical statistics, significant behavioral predictors were deduced from a logistic regression model that classified students and biopharma company employees as experts or novices on pH meter handling with 77% accuracy. Specifically, the game score and number of interactions in VR tasks requiring practical skills were found to be performance predictors. The study provides biopharma companies and academic institutions the possibility of assessing performance using an automatic, reliable, and simple alternative to traditional in-person assessment methods. Integrating the assessment into the training tool renders such laborious post-training assessments unnecessary.

Characterization of Eight Bacterial Biosensors for Microbial Diagnostic and Therapeutic Applications.

The engineering of microbial cells to produce and secrete therapeutics directly in the human body, known as advanced microbial therapeutics, is an exciting alternative to current drug delivery routes. These living therapeutics can be engineered to sense disease biomarkers and, in response, deliver a therapeutic activity. This strategy allows for precise and self-regulating delivery of a therapeutic that adapts to the disease state of the individual patient. Numerous sensing systems have been characterized for use in prokaryotes, but a very limited number of advanced microbial therapeutics have incorporated such sensors. We characterized eight different sensors that respond to physiologically relevant conditions and molecules found in the human body in the probiotic strain Escherichia coli Nissle 1917. The resulting sensors were characterized under aerobic and anaerobic conditions and were demonstrated to be functional under gut-like conditions using the nematode Caenorhabditis elegans as an in vivo model. We show for the first time how a biosensor is able to detect in vivo the bile acid-like molecule Δ4-dafachronic acid, a small molecule in C. elegans that regulates lifespan. Furthermore, we exemplify how bacterial sensors can be used to dynamically report on changes in the intestinal environment of C. elegans, by demonstrating the use of a biosensor able to detect changes in lactate concentrations in the gut lumen of individual C. elegans. The biosensors presented in this study allow for dynamic control of expression in vivo and represent a valuable tool in further developing advanced microbiome therapeutics.

Screening for effective cell-penetrating peptides with minimal impact on epithelial cells and gut commensals in vitro.

One of the biggest challenges for oral drug absorption is the epithelial barrier of the gastrointestinal tract. The use of cell-penetrating peptides (CPPs) to modulate the epithelial barrier function is known to be an effective strategy to improve drug absorption and bioavailability. In this study we compare side-by-side, 9 most promising CPPs to study their cytotoxicity (Cytotox Red dye staining) and cell viability (AlamarBlue staining) on epithelial cells and their effects on paracellular permeability of the intestinal barrier in vitro in a differentiated Caco-2 epithelial monolayer model. The data revealed that 4 out of 9 well-studied CPPs significantly improved Caco-2 paracellular permeability without compromising on cellular health. To assess the impact of CPPs on the human microbiota we studied the antimicrobial effects of the 4 effective CPPs from our permeation studies against 10 representative strains of the gut microbiota in vitro using microbroth dilution. Our data revealed that these 4 CPPs affected the growth of almost all tested commensal strains. Interestingly, we found that two synthetic CPPs (Shuffle and Penetramax) outperformed all the other CPPs in their ability to increase intestinal paracellular permeability at 50 µM and had only a small to moderate effect on the tested gut commensal strains. Based on these data Shuffle and Penetramax represent relevant CPPs to be further characterized in vivo for safe delivery of poorly absorbed therapeutics while minimizing negative impacts on the gut microbiota.

Arthroscopic Rotator Cuff Repair with a Fibrin Scaffold Containing Growth Factors and Autologous Progenitor Cells Derived From Humeral cBMA Improves Clinical Outcomes in High Risk Patients.

Purpose: To report the clinical outcomes after biologically augmented rotator cuff repair (RCR) with a fibrin scaffold derived from autologous whole blood and supplemented with concentrated bone marrow aspirate (cBMA) harvested at the proximal humerus. Methods: Patients who underwent arthroscopic RCR with biologic augmentation using a fibrin clot scaffold ("Mega- Clot") containing progenitor cells and growth factors from proximal humerus BMA and autologous whole blood between April 2015 and January 2018 were prospectively followed. Only high-risk patients in primary and revision cases that possessed relevant comorbidities or physically demanding occupation were included. Minimum follow-up for inclusion was 1 year. The visual analog score for pain (VAS), American Shoulder and Elbow Surgeons (ASES), Simple Shoulder Test (SST), Single Assessment Numerical Evaluation (SANE), and Constant-Murley scores were collected preoperatively and at final follow-up. In vitro analyses of the cBMA and fibrin clot using nucleated cell count, colony forming units, and live/dead assays were used to quantify the substrates. Results: Thirteen patients (56.9 ± 7.7 years) were included. The mean follow-up was 26.9 ± 17.7 months (n = 13). There were significant improvements in all outcome scores from the preoperative to the postoperative state: VAS (5.6 ± 2.5 to 3.1 ± 3.2; P < .001), ASES (42.0 ± 17.1 to 65.5 ± 30.6; P < .001), SST (3.2 ± 2.8 to 6.5 ± 4.7; P = .002), SANE (11.5 ± 15.6 to 50.3 ± 36.5; P < .001), and Constant-Murley (38.9 ± 17.5 to 58.1 ± 26.3; P < .001). Six patients (46%) had retears on postoperative MRI, despite all having improvements in pain and function except one. All failures were chronic rotator cuff tears, and all were revision cases except one (1.6 ± 0.5 previous RCRs). The representative sample of harvested cBMA showed an average of 28.5 ± 9.1 × 106 nucleated cells per mL. Conclusions: Arthroscopic rotator cuff repairs that are biologically augmented with a fibrin scaffold containing growth factors and autologous progenitor cells derived from autologous whole blood and humeral cBMA can improve clinical outcomes in primary, as well as revision cases in high-risk patients. However, the incidence of retears remains a concern in this population, demanding further improvements in biologic augmentation. Level of Evidence: IV, therapeutic case series.

A Novel Efficient L-Lysine Exporter Identified by Functional Metagenomics.

Lack of active export system often limits the industrial bio-based production processes accumulating the intracellular product and hence complexing the purification steps. L-lysine, an essential amino acid, is produced biologically in quantities exceeding two million tons per year; yet, L-lysine production is challenged by efficient export system at high titers during fermentation. To address this issue, new exporter candidates for efficient efflux of L-lysine are needed. Using metagenomic functional selection, we identified 58 genes encoded on 28 unique metagenomic fragments from cow gut microbiome library that improved L-lysine tolerance. These genes include a novel L-lysine transporter, belonging to a previously uncharacterized EamA superfamily, which is further in vivo characterized as L-lysine exporter using Xenopus oocyte expression system as well as Escherichia coli host. This novel exporter improved L-lysine tolerance in E. coli by 40% and enhanced yield, titer, and the specific production of L-lysine in an industrial Corynebacterium glutamicum strain by 7.8%, 9.5%, and 12%, respectively. Our approach allows the sequence-independent discovery of novel exporters and can be deployed to increase titers and productivity of toxicity-limited bioprocesses.

Effects of broad-spectrum antibiotics on the colonisation of probiotic yeast Saccharomyces boulardii in the murine gastrointestinal tract.

Mouse models are commonly used to study the colonisation profiles of microorganisms introduced to the gastrointestinal tract. Three commonly used mouse models include conventional, germ-free, and antibiotic-treated mice. However, colonisation resistance in conventional mice and specialised equipment for germ-free mice are usually limiting factors in their applications. In this study, we sought to establish a robust colonisation model for Saccharomyces boulardii, a probiotic yeast that has caught attention in the field of probiotics and advanced microbiome therapeutics. We characterised the colonisation of S. boulardii in conventional mice and mice treated with a cocktail of broad-spectrum antibiotics, including ampicillin, kanamycin, metronidazole and vancomycin. We found colonisation levels increased up to 10,000-fold in the antibiotic-treated mice compared to nonantibiotic-treated mice. Furthermore, S. boulardii was detected continuously in more than 75% of mice for 10 days after the last administration in antibiotic-treated mice, in contrast to in nonantibiotic-treated mice where S. boulardii was undetectable in less than 2 days. Finally, we demonstrated that this antibiotic cocktail can be used in two commonly used mouse strains, C57BL/6 and ob/ob mice, both achieving ~ 108 CFU/g of S. boulardii in faeces. These findings highlight that the antibiotic cocktail used in this study is an advantageous tool to study S. boulardii based probiotic and advanced microbiome therapeutics.

Orthogeriatric co-management: differences in outcome between major and minor fractures.

PURPOSE: Literature shows that orthogeriatric co-management improves the outcomes of patients with hip fractures. Corresponding research with more diverse fragility fracture groups is lacking. Therefore, an examination was performed prospectively as a 2 year-follow-up on an orthogeriatric co-managed ward, comparing relevant outcome parameters for major and minor fragility fractures. METHODS: All patients treated on an orthogeriatric co-managed ward from February 2014 to January 2015 were included and their injuries, orthogeriatric parameters such as the Barthel Index (BI), Parker Mobility Score (PMS) and place of residence (POR). Patients were separated into two groups of either immobilizing major (MaF) or non-immobilizing minor (MiF) fractures. 2 years later, a follow-up was conducted via telephone calls and questionnaires mailed to patients and/or their relatives. RESULTS: 740 (574 major vs. 166 minor injuries) patients were initially assessed, with a follow-up rate of 78.9%. The in-house, 1-year, and 2-year-mortality rates were 2.7, 27.4, and 39.2%, respectively. Mortality was significantly higher for MaF in the short term, but not after 2 years. On average, during the observation period, patients regained their BI by 36.7 points (95% CI: 33.80-39.63) and PMS was reduced by 1.4 points (95% CI: 1.16-1.68). No significant differences were found in the readmission rate, change in BI, PMS or POR between the MaF and MiF groups. CONCLUSION: The relevance of orthogeriatric treatment to improving functional and socioeconomic outcomes was confirmed. The similarity of the results from both fracture groups emphasizes the need for a multidisciplinary approach also for minor fractures.

Semitendinosus vs Gracilis Grafts With 1- vs 2-Tunnel Techniques for Coracoclavicular Ligament Reconstruction: A Biomechanical Study.

BACKGROUND: Despite the evolution of acromioclavicular joint surgery to a more anatomic coracoclavicular (CC) ligament reconstruction, no definitive guidance regarding the number and position of bone tunnels in the clavicle, as well as the ideal graft choice, is established. PURPOSE/HYPOTHESIS: The purpose of this study was to biomechanically compare the reconstruction of the CC ligament complex between gracilis- and semitendinosus-tendon grafts in 1- and 2-tunnel techniques. It was hypothesized that the gracilis tendon graft will provide comparable primary stability in both tunnel techniques while utilizing a smaller tunnel diameter. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 24 cadaveric shoulders (13 men, 11 women; 66 ± 7.5 years) were randomly allocated to 4 repair groups: gracilis with 1 tunnel (GT-1), gracilis with 2 tunnels (GT-2), semitendinosus with 1 tunnel (ST-1), and semitendinosus with 2 tunnels (ST-2). First, specimens were tested for native anterior, posterior, and superior translations. Then, specimens were randomly assigned to 1 of the 4 CC reconstruction groups before undergoing the same testing, followed by cyclic loading and load to failure (LTF). RESULTS: The GT-2 reconstruction demonstrated significantly less translation when compared with ST-2 in anterior (P = .024) and posterior (P = .048) directions. GT-1 and ST-2 both showed significantly less translation than ST-1 in anterior and superior directions (P < .001). All reconstructions demonstrated less superior translation compared with native testing, with GT-1 and ST-2 significantly less than ST-1 (P < .001). There were no significant differences for peak displacement and LTF between groups. CONCLUSION: Gracilis tendon grafts using a 1- or 2-tunnel technique for CC ligament reconstruction provided comparable translation, displacement, and LTF as corresponding semitendinosus grafts. Therefore, the gracilis tendon should be considered as a biomechanical equivalent graft choice for the reconstruction of the CC ligament complex. CLINICAL RELEVANCE: In a cadaveric model, the gracilis tendon demonstrated adequate fixation with minimal translation in CC ligament reconstruction while utilizing smaller diameter bone tunnels, which may help minimize the risk of complications such as loss of reduction and fracture.