NOS inhibition reverses TLR2-induced chondrocyte dysfunction and attenuates age-related osteoarthritis.

Osteoarthritis (OA) is a joint disease featuring cartilage breakdown and chronic pain. Although age and joint trauma are prominently associated with OA occurrence, the trigger and signaling pathways propagating their pathogenic aspects are ill defined. Following long-term catabolic activity and traumatic cartilage breakdown, debris accumulates and can trigger Toll-like receptors (TLRs). Here we show that TLR2 stimulation suppressed the expression of matrix proteins and induced an inflammatory phenotype in human chondrocytes. Further, TLR2 stimulation impaired chondrocyte mitochondrial function, resulting in severely reduced adenosine triphosphate (ATP) production. RNA-sequencing analysis revealed that TLR2 stimulation upregulated nitric oxide synthase 2 (NOS2) expression and downregulated mitochondria function-associated genes. NOS inhibition partially restored the expression of these genes, and rescued mitochondrial function and ATP production. Correspondingly, Nos2-/- mice were protected from age-related OA development. Taken together, the TLR2-NOS axis promotes human chondrocyte dysfunction and murine OA development, and targeted interventions may provide therapeutic and preventive approaches in OA.

Autologous bone grafting in combination with autologous chondrocyte implantation yields favourable outcomes in the treatment of osteochondral defects of the knee: A systematic literature review.

PURPOSE: This study addresses the gap in the current literature by evaluating the combined treatment of autologous bone grafting and autologous chondrocyte implantation (ABCI) for osteochondral defects of the knee. It aims to evaluate clinical outcomes against methodological quality and to summarize histological results and surgical techniques. METHODS: A thorough search was conducted across Pubmed, Cochrane and Embase databases. Studies reporting clinical outcomes of ABCI for osteochondral defects of the knee were included. Patient-reported outcome measures (PROMs), failure rates, methodological quality and potential conflicts of interest were evaluated. Histological results and surgical techniques were summarized. RESULTS: Eighteen studies with 344 analyzed patients met the eligibility criteria for inclusion. All studies showed a significant improvement (p < 0.05) across different PROMs (subjective International Knee Documentation Committee score, Cincinnati Knee Rating System, Visual Analogue Scale, Lysholm Score, Tegner Activity Scale, Knee injury and Osteoarthritis Outcome Score and Knee Society Score) compared to the preoperative status. Failure rates ranged from 0% to 17.6%, with a mean follow-up of 73.2 months (range: 9.0-143.6 months). Methodological quality was low to medium, including only one comparative study. Six studies reviewed reported a potential conflict of interest. The histological assessment showed effective bonding between autologous chondrocytes and bone graft. A large degree of variability was observed in the operative technique used. CONCLUSION: The current literature suggests that ABCI yields good clinical outcomes at mid- to long-term follow-up with favourable histological results for osteochondral defects of the knee. However, future research should focus on high-quality comparative studies to better guide treatment choices. Introducing ABCI as the standard abbreviation may enhance clarity in future research. LEVEL OF EVIDENCE: Level IV.

No significant difference in skin contamination during anterior cruciate ligament reconstruction with and without preoperative skin cleaning.

PURPOSE: This prospective study aimed to assess whether preoperative antiseptic skin cleansing reduces bacterial contamination and surgical site infections (SSI) following anterior cruciate ligament reconstruction (ACLR). We hypothesized that antiseptic cleaning would lower bacterial load, reducing contamination and early infections. METHODS: One hundred and nineteen patients scheduled for ACLR were included in this prospective, nonrandomized study. Individuals were divided into two groups. Patients in the intervention group applied octenisan® wash lotion daily for three days before surgery and used the wash solution instead of their usual shower gel. Additionally, they swiped their leg with octenisan® soaked gloves on the morning of the operation. The control group followed their usual wash routine with no specific instructions. Fluid samples were taken before surgery from the irrigation bag and at 15-min intervals from the reservoir of the sterile surgical drape during the procedure. Suture material used for the ACL graft and meniscus repair were also collected for testing. The samples were subjected to a 14-day incubation period. Follow-up included outpatient visits at 6 weeks, 12 weeks and 6 months with a final evaluation at 12 months. RESULTS: Contamination rates showed no significant difference between the control and intervention groups. The mean contamination rate in the control group was 6.4% (n = 22) and 6.6% (n = 24) in the intervention group (p = 0.28). At 12-month follow-up, 110 out of 119 participants were included (52 control, 58 intervention). T tests for age (p = 0.19), BMI (p = 0.66), and surgery duration (p = 0.38) showed no significant differences. No early SSI were observed in either group postoperatively. CONCLUSION: Our results indicate that the use of antiseptic wash lotion and gloves does not influence the risk of bacterial contamination during surgery. LEVEL OF EVIDENCE: Level III.

Management after acute injury of the anterior cruciate ligament (ACL). Part 3: Recommendation on surgical treatment.

PURPOSE: The aim of this consensus project was to give recommendations regarding surgical treatment of the anterior cruciate ligament (ACL) injured patient. METHODS: For this consensus process, an expert, steering and rating group was formed. In an initial online meeting, the steering group, together with the expert group, formed various key topic complexes for which multiple questions were formulated. For each key topic, a structured literature search was performed by the steering group. The results of the literature review were sent to the rating group with the option to give anonymous comments until a final consensus voting was performed. Sufficient consensus was defined as 80% agreement. RESULTS: During this consensus process, 30 topics regarding the surgical management and technique of ACL reconstruction were identified. The literature search for each key question resulted in 30 final statements. Of these 30 final statements, all achieved consensus. CONCLUSIONS: This consensus process has shown that surgical treatment of ACL injury is a complex process. Various surgical factors influence patient outcomes. The proposed treatment algorithm can be used as a decision aid for the surgeon. LEVEL OF EVIDENCE: Level V.

An evaluation of a novel method for the MRI-based assessment of Caton-Deschamps index in the Knee.

INTRODUCTION: The radiographical assessment of patella height has historically been performed using X-Ray. The aim of this study was to evaluate a new method for the assessment of patella height using MRI and to assess the correlation with the X-Ray based assessment. MATERIALS AND METHODS: 159 patients who had both lateral radiographs and MRI images were included. Parameters measured included traditional radiographical CDI, MRI-based CDI, and TT-TG distance. On the basis of the TT-TG, the patients were divided into 2 groups. Two different methods were used to assess CDI using MRI: using a single slice image, and an alternative technique using two different cross-sectional images. The correlation of the two measurement methods was assessed using Pearson's correlation coefficient. The intraclass correlation coefficient (ICC) was determined from the measurements of the two investigators. RESULTS: The average TT-TG distance was 11.6 mm (± 4.6). In patients with a TT-TG < 15 mm, both measurement methods showed comparable correlation with measurements on X-Ray. In patients with a TT-TG of > 15 the the new cross-sectional imaging method showed higher correlation with traditional X-Ray assessment compared to CDI assessment using the traditional single slice method (r = 0.594, p < 0.001 vs. r = 0.302, p = 0.055). CONCLUSIONS: The assessment of CDI on MRI using a cross-sectional imaging method has a better correlation with traditional X-Ray assessment of CDI than single-slice assessment. This is particularly true in patients with elevated TT-TG and as such should be preferentially used in the assessment of Patellar height in this cohort.

High-fat, sucrose and salt-rich diet during rat spermatogenesis lead to the development of chronic kidney disease in the female offspring of the F2 generation.

Effects of feeding male rats during spermatogenesis a high-fat, high-sucrose and high-salt diet (HFSSD) over two generations (F0 and F1) on renal outcomes are unknown. Male F0 and F1 rats were fed either control diet (F0CD+F1CD) or HFSSD (F0HD+F1HD). The outcomes were glomerular filtration rate and urinary albumin excretion in F1 and F2 offspring. If both outcomes were altered a morphological and molecular assessment was done. F2 offspring of both sexes had a decreased GFR. However, increased urinary albumin excretion was only observed in female F2 F0HD+F1HD offspring compared with controls. F0HD+F1HD female F2 offspring developed glomerulosclerosis (+31%; p < .01) and increased renal interstitial fibrosis (+52%; p < .05). RNA sequencing followed by qRT-PCR validation showed that four genes (Enpp6, Tmem144, Cd300lf, and Actr3b) were differentially regulated in the kidneys of female F2 offspring. lncRNA XR-146683.1 expression decreased in female F0HD+F1HD F2 offspring and its expression was (r = 0.44, p = .027) correlated with the expression of Tmem144. Methylation of CpG islands in the promoter region of the Cd300lf gene was increased (p = .001) in female F2 F0HD+F1HD offspring compared to controls. Promoter CpG island methylation rate of Cd300lf was inversely correlated with Cd300lf mRNA expression in F2 female offspring (r = -0.483, p = .012). Cd300lf mRNA expression was inversely correlated with the urinary albumin-to-creatinine ratio in female F2 offspring (r = -0.588, p = .005). Paternal pre-conceptional unhealthy diet given for two generations predispose female F2 offspring to chronic kidney disease due to epigenetic alterations of renal gene expression. Particularly, Cd300lf gene promotor methylation was inversely associated with Cd300lf mRNA expression and Cd300lf mRNA expression itself was inversely associated with urinary albumin excretion in F2 female offspring whose fathers and grandfathers got a pre-conceptional unhealthy diet.

Management after acute injury of the anterior cruciate ligament (ACL), part 2: management of the ACL-injured patient.

PURPOSE: The aim of this consensus project was to create a treatment algorithm for the management of the ACL-injured patient which can serve as an aid in a shared decision-making process. METHODS: For this consensus process, a steering and a rating group were formed. In an initial face-to-face meeting, the steering group, together with the expert group, formed various key topic complexes for which various questions were formulated. For each key topic, a structured literature search was performed by the steering group. The results of the literature review were sent to the rating group with the option to give anonymous comments until a final consensus voting was performed. Sufficient consensus was defined as 80% agreement. RESULTS: During this consensus process, 15 key questions were identified. The literature search for each key question resulted in 24 final statements. Of these 24 final statements, all achieved consensus. CONCLUSIONS: This consensus process has shown that ACL rupture is a complex injury, and the outcome depends to a large extent on the frequently concomitant injuries (meniscus and/or cartilage damage). These additional injuries as well as various patient-specific factors should play a role in the treatment decision. The present treatment algorithm represents a decision aid within the framework of a shared decision-making process for the ACL-injured patient. LEVEL OF EVIDENCE: Level V.

Bacterial contamination of irrigation fluid and suture material during ACL reconstruction and meniscus surgery : Low infection rate despite increasing contamination over surgery time.

PURPOSE: During knee arthroscopy, irrigation fluid from the surgical site accumulates in the sterile reservoir. Whether these fluid collections and also suture material used during the arthroscopic surgical processes show bacterial contamination over time during surgery remains unclear. The purpose of this study was to determine this contamination rate and to analyze its possible influence on postoperative infection. MATERIALS AND METHODS: In this study, 155 patients were included. Fifty-eight underwent reconstruction of the anterior cruciate ligament (ACL), 63 meniscal surgery and 34 patients combined ACL reconstruction and meniscus repair. We collected pooled samples of irrigation fluid from the reservoir on the sterile drape every 15 min during the surgery. In addition, we evaluated suture material of ACL graft and meniscus repair for bacterial contamination. Samples were sent for microbiological analysis, incubation time was 14 days. All patients were seen in the outpatient department 6, 12 weeks and 12 months postoperatively and examined for clinical signs of infection. RESULTS: A strong statistical correlation (R2 = 0.81, p = 0.015) was found between an advanced duration of surgery and the number of positive microbiological findings in the accumulated fluid. Suture and fixation material showed a contamination rate of 28.4% (29 cases). Despite the high contamination rate, only one infection was found in the follow-up examinations, caused by Staphylococcus lugdunensis. CONCLUSION: Since bacterial contamination of accumulated fluid increases over time the contact with the fluid reservoirs should be avoided. LEVEL OF EVIDENCE: IV.

The subjective knee value is a valid single-item survey to assess knee function in common knee disorders.

INTRODUCTION: The patient's perspective plays a key role in judging the effect of knee disorders on physical function. We have introduced the Subjective Knee Value (SKV) to simplify the evaluation of individual's knee function by providing one simple question. The purpose of this prospective study was to validate the SKV with accepted multiple-item knee surveys across patients with orthopaedic knee disorders. MATERIALS AND METHODS: Between January through March 2020, consecutive patients (n = 160; mean age 51 ± 18 years, range from 18 to 85 years, 54% women) attending the outpatient clinic for knee complaints caused by osteoarthritis (n = 69), meniscal lesion (n = 45), tear of the anterior cruciate ligament (n = 23) and focal chondral defect (n = 23) were invited to complete a knee-specific survey including the SKV along with the Knee Injury Osteoarthritis Outcome Score (KOOS) and the International Knee Documentation Committee subjective knee form (IKDC-S). The Pearson correlation coefficient was used to evaluate external validity between the SKV and each patient-reported outcome measure (PROM) separately. Furthermore, patient's compliance was assessed by comparing responding rates. RESULTS: Overall, the SKV highly correlated with both the KOOS (R = 0.758, p < 0.05) and the IKDC-S (R = 0.802, p < 0.05). This was also demonstrated across all investigated diagnosis- and demographic-specific (gender, age) subgroups (range 0.509-0.936). No relevant floor/ceiling effects were noticed. The responding rate for the SKV (96%) was significantly higher when compared with those for the KOOS (81%) and the IKDC-S (83%) (p < 0.05). CONCLUSION: At baseline, the SKV exhibits acceptable validity across all investigated knee-specific PROMs in a broad patient population with a wide array of knee disorders. The simplified survey format without compromising the precision to evaluate individual's knee function justifies implementation in daily clinical practice. LEVEL OF EVIDENCE: II, cohort study (diagnosis).

[Influence of the COVID-19 pandemic on the physiotherapeutic and medical follow-up care after reconstructions of the anterior cruciate ligament]. / Einfluss der COVID-19-Pandemie auf die physiotherapeutische und ärztliche Nachsorge nach Rekonstruktionen des vorderen Kreuzbandes.

BACKGROUND: The aim of this study was to examine the influence of the COVID-19(coronavirus disease 2019) pandemic on the medical and physiotherapeutic follow-up care after reconstructions of the anterior cruciate ligament. METHODS: 116 patients (72 men and 44 women) who had received a reconstruction of the anterior cruciate ligament between September 2019 and December 2020 were included in this study. These patients were divided into two groups depending on the date of surgery: one group that had received surgery in 2019 before the COVID-19 pandemic and one group that had undergone surgery in 2020 and was directly affected by the COVID-19 pandemic. All patients were interviewed using a standardized questionnaire regarding the influence of the coronavirus pandemic on the medical and physiotherapeutic follow-up care, as well as the moment of return to work. In addition, the range of motion of the respective knee 3 months postoperatively was analysed based on physical examination. RESULTS: At 3 months postoperatively, patients who had undergone surgery in 2020 showed a clear trend towards a higher frequency of extension deficits of ≥ 5° (18.8% vs. 4.3%, p = 0.097) or an inability to bend the knee ≥ 120° (23.3% vs. 10%, p = 0.197) compared to those who had received surgery in 2019. Patients who had undergone surgery in 2020 reported significantly longer delays for appointments, a higher number of futile attempts to get an appointment and a higher number of cancelled appointments, regarding both medical and physiotherapeutic follow-up care. 34.9% of the patients who received surgery in 2020 indicated that they were able to reduce the duration of their sick leave due to the increased possibilities of working in a home-office situation during the pandemic. Alternative treatment options due to the pandemic were offered by 13.3% of the physiotherapists and 12.2% of the physicians. CONCLUSION: Although the physiotherapeutic and medical follow-up care was not directly affected by a "lockdown", the pandemic led to significant restrictions, which are also reflected in a clear trend towards worse clinical outcomes. Consequently, a further expansion of alternative treatment options, which were only offered by 12-13% of practices and that are presumably of comparable relevance for various other diseases, is needed.

Posterior laxity increases over time after PCL reconstruction.

PURPOSE: Restoration of posterior tibial translation (PTT) after reconstruction of the posterior cruciate ligament (PCL) is deemed necessary to restore physiological knee kinematics. However, current surgical techniques have failed to show a complete reduction of posterior laxity. It was hypothesized that early postoperative PTT increases over time. METHODS: The study comprised of 46 patients (10 female, 36 male; 30 ± 9 years), who underwent PCL reconstruction in a single-surgeon series. Patients were evaluated by bilateral stress radiographs in a prospective manner preoperatively; at 3, 6, 12 and 24 months; and at a final follow-up (FFU) of at least 5 years. Covariants included age, gender, BMI, tibial slope (TS) and the number of operated ligaments. Two blinded observers reviewed all radiographs, evaluating the TS and the posterior tibial translation. RESULTS: All patients were evaluated at a mean final follow-up of 102 (range 65-187) months. Mean side-to-side difference of the PTT significantly improved from preoperative to 3-month postoperative values (10.9 ± 3.1 vs. 3.6 ± 3.8 mm; P < 0.0001). The PTT increased to 4.6 ± 3.7 mm at 6 months, to 4.8 ± 3.3 mm at 12 months, to 4.8 ± 3.2 mm at 24 months, to 5.4 ± 3.4 mm at FFU. Consequently, there was a significant increase of PTT between 3-month and final follow-up (3.6 ± 3.8 vs. 5.4 ± 3.4 mm; P = 0.02). Flattening of the TS resulted in a significantly higher PTT compared to subjects with a high TS at 24 months and FFU. There was no significant influence by BMI, age, gender and the number of operated ligaments. CONCLUSIONS: Early results after PCL reconstruction seem promising as posterior tibial translation is significantly improved. However, there is a significant increase in PTT from early postoperative values to the final follow-up of at least 5 years. This is particularly notable in patients with flattening of the TS. As a consequence, surgeons and patients need to be aware that initial posterior stability should not be equated with the final outcome. LEVEL OF EVIDENCE: Cohort study, Level III.

Treatment of osteochondral defects: chondrointegration of metal implants improves after hydroxyapatite coating.

PURPOSE: The treatment of osteochondral defects in joint cartilage remains challenging due to its limited repair capacity. This study presents a metallic osteochondral plug with hydroxyapatite (HA)-coated cap edges for improved implant-tissue contact. The hypothesis was that improved attachment prevents from synovial fluid-influx and thereby avoids osteolysis and resulting implant instability. METHODS: In total, 24 female, adult sheep were randomized into three groups. All animals received an Episealer®-implant in the medial condyle of the right knee. The implants were coated with two different HA versions or uncoated (control group). After 12 weeks, the implant-tissue connections were analysed radiologically and histologically. RESULTS: In general, the groups with the coated cap edges showed a better quality of tissue connection to the implant. The occurrence of gaps between tissue and implant was more seldom, the binding of calcified and hyaline cartilage to the cap was significantly better than in the uncoated group. A histomorphometrically measured lower amount of void space in these groups compared to the group with the uncoated edges confirmed that. CONCLUSIONS: The hypothesis of a tighter cartilage bone contact was confirmed. The HA coating of the implant's cap edges resulted in better adherence of cartilage to the implant, which was not previously reported. In conclusion, this led to a better contact between implant and cartilage as well as neighbouring bone. In clinical routine, joint fluid is aggressive, penetrates through cartilage rifts, and promotes osteolysis and loosening of implants. The observed sealing effect will act to prevent joint fluid to get access to the implant-tissue interfaces. Joint fluid is aggressive, can cause osteolysis, and can, clinically cause pain. These effects are liable to decrease with these findings and will further the longevity of these osteochondral implants.

Functional Characterization of Three Specific Acyl-Coenzyme A Synthetases Involved in Anaerobic Cholesterol Degradation in Sterolibacterium denitrificans Chol1S.

The denitrifying betaproteobacterium Sterolibacterium denitrificans Chol1S catabolizes steroids such as cholesterol via an oxygen-independent pathway. It involves enzyme reaction sequences described for aerobic cholesterol and bile acid degradation as well as enzymes uniquely found in anaerobic steroid-degrading bacteria. Recent studies provided evidence that in S. denitrificans, the cholest-4-en-3-one intermediate is oxygen-independently oxidized to Δ4-dafachronic acid (C26-oic acid), which is subsequently activated by a substrate-specific acyl-coenzyme A (acyl-CoA) synthetase (ACS). Further degradation was suggested to proceed via unconventional ß-oxidation, where aldolases, aldehyde dehydrogenases, and additional ACSs substitute for classical ß-hydroxyacyl-CoA dehydrogenases and thiolases. Here, we heterologously expressed three cholesterol-induced genes that putatively code for AMP-forming ACSs and characterized two of the products as specific 3ß-hydroxy-Δ5-cholenoyl-CoA (C24-oic acid)- and pregn-4-en-3-one-22-oyl-CoA (C22-oic acid)-forming ACSs, respectively. A third heterologously produced ATP-dependent ACS was inactive with C26-, C24-, or C22-oic-acids but activated 3aα-H-4α-(3'propanoate)-7aß-methylhexahydro-1,5-indanedione (HIP) to HIP-CoA, a rather late intermediate of aerobic cholesterol degradation that still contains the CD rings of the sterane skeleton. This work provides experimental evidence that anaerobic steroid degradation proceeds via numerous alternate CoA-ester-dependent or -independent enzymatic reaction sequences as a result of aldolytic side chain and hydrolytic sterane ring C-C bond cleavages. The aldolytic side chain degradation pathway comprising highly exergonic ACSs and aldehyde dehydrogenases is considered to be essential for driving the unfavorable oxygen-independent C26 hydroxylation forward.IMPORTANCE The biological degradation of ubiquitously abundant steroids is hampered by their low solubility and the presence of two quaternary carbon atoms. The degradation of cholesterol by aerobic Actinobacteria has been studied in detail for more than 30 years and involves a number of oxygenase-dependent reactions. In contrast, much less is known about the oxygen-independent degradation of steroids in denitrifying bacteria. In the cholesterol-degrading anaerobic model organism Sterolibacterium denitrificans Chol1S, initial evidence has been obtained that steroid degradation proceeds via numerous alternate coenzyme A (CoA)-ester-dependent/independent reaction sequences. Here, we describe the heterologous expression of three highly specific and characteristic acyl-CoA synthetases, two of which play key roles in the degradation of the side chain, whereas a third one is specifically involved in the B ring degradation. The results obtained shed light into oxygen-independent steroid degradation comprising more than 40 enzymatic reactions.

Kyrpidia spormannii sp. nov., a thermophilic, hydrogen-oxidizing, facultative autotroph, isolated from hydrothermal systems at São Miguel Island, and emended description of the genus Kyrpidia.

A Gram-stain-positive, rod-shaped, non-motile, spore-forming bacterium, strain EA-1T, was isolated from hydrothermal sediment samples from the Azores (São Miguel, Portugal). 16S rRNA gene sequence analysis of the isolated bacterium revealed a phylogenetic affiliation with the genus Kyrpidia. The sequence similarity of the five 16S rRNA gene copies to its closest relative, Kyrpidia tusciae, ranged from 97.79 to 97.85 %. The in silico estimate of DNA-DNA hybridization was 56.0 %. The dominant fatty acids of the novel isolate were anteiso-C17 : 0 (49.9 %), iso-C17 : 0 (23.0 %) and iso-C16 : 0 (13.3 %), while the quinone detected was menaquinone MK-7. Analysis of polar lipids identified phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and additional unidentified compounds comprising two glycolipids, two phospholipids and two lipids. The presence of meso-diaminopimelic acid in the peptidoglycan and mannose, arabinose and ribose in the cell wall of strain EA-1T were detected. The strain was able to grow heterotrophically as well as autotrophically with carbon dioxide as the sole carbon source and with hydrogen and oxygen as electron donor and acceptor, respectively. Based on its chemotaxonomic, physiological and genomic characteristics, the new strain is considered to represent a novel species within the genus Kyrpidia, for which the name Kyrpidiaspormannii sp. nov. is proposed. The type strain is strain EA-1T (=DSM 106492T=CCOS1194T).

High tibial slope correlates with increased posterior tibial translation in healthy knees.

PURPOSE: Notwithstanding the importance of the tibial slope (TS) for anterior tibial translation, little information is available regarding the implications on posterior laxity, particularly in healthy subjects. It was hypothesized that increased TS is associated with decreased posterior tibial translation (PTT) in healthy knees. METHODS: A total of 124 stress radiographs of healthy knees were enrolled in this study. Tibial slope and the posterior tibial translation were evaluated using a Telos device with a 150-N force at 90° of knee flexion. Two blinded observers reviewed independently on two different occasions. RESULTS: One hundred and twenty-four patients [35 females and 89 males; 41 (range 18-75) years] were enrolled in this study, with a mean PTT of 2.8 mm (±1.9 mm; range 0-8 mm) and a mean TS of 8.6° (±2.6°; range 1°-14°). Pearson correlation showed a significant correlation between the PTT and TS in the overall patient cohort (P < 0.0001) with r = 0.76 and R 2 = 0.58. There was no statistical difference between female and male patients regarding the PTT or the TS. Subgrouping of the patient cohort (four groups with n = 31) according to their TS (groups I < 7°; II = 7°-8.5°; III = 9°-10.5°; IV ≥ 11°) revealed significant differences between each subgroup, respectively. Furthermore, there was a weak but significant correlation between age and PTT (P = 0.004, r = 0.26). CONCLUSION: In addition to the substantial variance in tibial slope and posterior laxity among healthy knees, high tibial slope significantly correlates with increased posterior tibial translation. Increasing age is further associated with a greater magnitude of posterior tibial translation. Consequently, knowledge of the tibial slope facilitates simple estimation of posterior knee laxity, which is mandatory for PCL reconstruction and knee arthroplasty.

Degenerative changes after posterior cruciate ligament reconstruction are irrespective of posterior knee stability: MRI-based long-term results.

INTRODUCTION: Posterior cruciate ligament reconstruction (PCLR) is advocated to prevent an early onset of osteoarthritis. We hypothesized that posterior instability after PCLR correlates with degenerative changes. MATERIALS AND METHODS: MRIs of 42 (12 female/30 male; 39 ± 9 years) patients were enrolled with a minimum 5-year follow-up (FFU) after PCLR. In addition, 25 contralateral and 15 follow-up MRIs (12 months after baseline) were performed. Degenerative changes were graded using WORMS. Posterior tibial translation (PTT) was measured using posterior stress radiographs. Outcome parameters included WORMS/cartilage subscore for the whole joint, patellofemoral (PFJ), medial (MFTJ), and lateral femorotibial joint (LFTJ). RESULTS: Final follow-up was 101 (range 68-168) months. WORMS reached 41.5 [18.5-56.8]. Regional WORMS for PFJ was significantly higher than MFTJ and LFTJ. Cartilage subscore yielded 7 [2.8-15]. MFTJ and PFJ were significantly higher than LFTJ. Primary outcome parameters were significantly higher than the contralateral knee (P < 0.0001) and significantly increased within 12 months (P = 0.0002). There was a significant correlation between the intraoperative degree of cartilage injury and WORMS (P < 0.0001 with r = 0.64) and between the number of previous surgery and the cartilage subscore (P = 0.03 with r = 0.32). Meniscal surgery led to a significantly higher WORMS (P = 0.035). Combined risk models revealed that women below the mean age had significantly lower WORMS (P = 0.001) and cartilage subscores (P = 0.003). CONCLUSIONS: Patients undergo degenerative changes after PCLR, which are significantly higher compared to the contralateral knee. These occur predominantly at PFJ/MFTJ and are irrespective of posterior stability. Concomitant meniscus/cartilage injuries and a high number of previous surgeries are further risk factors.

Biomechanical performance of the Actifit® scaffold is significantly improved by selection of irrigation fluid.

PURPOSE: Clinical reports on meniscal scaffolds seem promising, albeit relatively paucity exists regarding their biomechanical behavior. The aim of the study is to delineate the impact of differing suture materials and the type as well as the temperature of the irrigation fluid on the pull-out strength of a polyurethane meniscal scaffold (Actifit®). MATERIALS AND METHODS: 128 specimens were utilized with horizontal sutures and uniaxial load-to-failure testing was performed. We compared two different suture materials-polydioxanone (PDS) and non-absorbable, braided polyester sutures (NABP)-as well as two common irrigation fluids-lactated Ringer's and electrolyte-free, hypotonic Mannitol-Sorbitol. All specimens were further evaluated according to two different temperatures [room temperature (20 °C) and near-core body temperature (37 °C)]. RESULTS: Mean load-to-failure was 53.3 ± 6.5 N. There was no significant difference between the NABP and the PDS group. Ringer group showed a significantly higher load-to-failure compared to Purisole (P = .0002). This was equivalent for both PDS (P = .0008) and NABP sutures (P = .0008). Significantly higher failure loads could be established for the 37° group (P = .041); yet, this difference was neither confirmed for the PDS or in in the NABP subgroup. Only the subgroup using Purisole at 37° showed significantly higher failure loads compared to 20° (P = .017). CONCLUSIONS: This study underlines the potential to improve pull-out strength during implantation of an Actifit® scaffold by alteration of the type of irrigation fluid. Lactated Ringer solution provided the highest construct stability in regard to load-to-failure testing and should be considered whenever implantation of a polyurethane meniscal scaffold is conducted.

A patchwork pathway for oxygenase-independent degradation of side chain containing steroids.

The denitrifying betaproteobacterium Sterolibacterium denitrificans serves as model organism for studying the oxygen-independent degradation of cholesterol. Here, we demonstrate its capability of degrading various globally abundant side chain containing zoo-, phyto- and mycosterols. We provide the complete genome that empowered an integrated genomics/proteomics/metabolomics approach, accompanied by the characterization of a characteristic enzyme of steroid side chain degradation. The results indicate that individual molybdopterin-containing steroid dehydrogenases are involved in C25-hydroxylations of steroids with different isoprenoid side chains, followed by the unusual conversion to C26-oic acids. Side chain degradation to androsta-1,4-diene-3,17-dione (ADD) via aldolytic C-C bond cleavages involves acyl-CoA synthetases/dehydrogenases specific for the respective 26-, 24- and 22-oic acids/-oyl-CoAs and promiscuous MaoC-like enoyl-CoA hydratases, aldolases and aldehyde dehydrogenases. Degradation of rings A and B depends on gene products uniquely found in anaerobic steroid degraders, which after hydrolytic cleavage of ring A, again involves CoA-ester intermediates. The degradation of the remaining CD rings via hydrolytic cleavage appears to be highly similar in aerobic and anaerobic bacteria. Anaerobic cholesterol degradation employs a composite repertoire of more than 40 genes partially known from aerobic degradation in gammaproteobacteria/actinobacteria, supplemented by unique genes that are required to circumvent oxygenase-dependent reactions.

25-Hydroxyvitamin†D3 Synthesis by Enzymatic Steroid Side-Chain Hydroxylation with Water.

The hydroxylation of vitamin D3 (VD3, cholecalciferol) side chains to give 25-hydroxyvitamin D3 (25OHVD3) is a crucial reaction in the formation of the circulating and biologically active forms of VD3 . It is usually catalyzed by cytochrome P450 monooxygenases that depend on complex electron donor systems. Cell-free extracts and a purified Mo enzyme from a bacterium anaerobically grown with cholesterol were employed for the regioselective, ferricyanide-dependent hydroxylation of VD3 and proVD3 (7-dehydrocholesterol) into the corresponding tertiary alcohols with greater than 99 % yield. Hydroxylation of VD3 strictly depends on a cyclodextrin-assisted isomerization of VD3 into preVD3 , the actual enzymatic substrate. This facile and robust method developed for 25OHVD3 synthesis is a novel example for the concept of substrate-engineered catalysis and offers an attractive alternative to chemical or O2 /electron-donor-dependent enzymatic procedures.

Reliable assessment of liver function using LiMAx.

BACKGROUND: (13)C-liver function breath tests can facilitate the assessment of hepatic function in-vivo and may help surgeons to identify candidates for safe liver surgery. However, their acceptance into clinical practice is dependent on evaluation of technical efficacy and repeatability. The aims of this study were to evaluate the within-subject repeatability of the LiMAx (maximum liver function capacity) test in healthy individuals and in surgical patients to determine liver function in the perioperative workup. MATERIAL AND METHODS: The LiMAx test, which is based on intravenous injection of (13)C-methacetin at a dosage of 2 mg/kg body weight was performed in eighty-six healthy subjects to determine a reference range. Twenty-four subjects underwent repeat LiMAx testing the following day to assess within-subject repeatability. Twenty-one patients undergoing elective extra-abdominal surgery under general anesthesia (GA group) received pre- and post-operative examinations. RESULTS: The normal range of LiMAx was found to be 430 ± 86 µg/kg/h and revealed a one-sided cut-off value of 315 µg/kg/h. The intraclass correlation coefficient of the repeat LiMAx tests was 0.85 (95% confidence interval 0.69-0.93) in the control group and 0.81 (95% confidence interval 0.60-0.92) in the group of patients with GA. CONCLUSIONS: The LiMAx test shows excellent reproducibility in subjects with normal liver function. GA has no effect on test results.