The N1 domain of the peroxisomal AAA-ATPase Pex6 is required for Pex15 binding and proper assembly with Pex1.

The heterohexameric ATPases associated with diverse cellular activities (AAA)-ATPase Pex1/Pex6 is essential for the formation and maintenance of peroxisomes. Pex1/Pex6, similar to other AAA-ATPases, uses the energy from ATP hydrolysis to mechanically thread substrate proteins through its central pore, thereby unfolding them. In related AAA-ATPase motors, substrates are recruited through binding to the motor's N-terminal domains or N terminally bound cofactors. Here, we use structural and biochemical techniques to characterize the function of the N1 domain in Pex6 from budding yeast, Saccharomyces cerevisiae. We found that although Pex1/ΔN1-Pex6 is an active ATPase in vitro, it does not support Pex1/Pex6 function at the peroxisome in vivo. An X-ray crystal structure of the isolated Pex6 N1 domain shows that the Pex6 N1 domain shares the same fold as the N-terminal domains of PEX1, CDC48, and NSF, despite poor sequence conservation. Integrating this structure with a cryo-EM reconstruction of Pex1/Pex6, AlphaFold2 predictions, and biochemical assays shows that Pex6 N1 mediates binding to both the peroxisomal membrane tether Pex15 and an extended loop from the D2 ATPase domain of Pex1 that influences Pex1/Pex6 heterohexamer stability. Given the direct interactions with both Pex15 and the D2 ATPase domains, the Pex6 N1 domain is poised to coordinate binding of cofactors and substrates with Pex1/Pex6 ATPase activity.

Efficacy of a Dual-Epitope Dendritic Cell Vaccine as Part of Combined Immunotherapy for HER2-Expressing Breast Tumors.

Previous work from our group and others has shown that patients with breast cancer can generate a T cell response against specific human epidermal growth factor 2 (HER2) epitopes. In addition, preclinical work has shown that this T cell response can be augmented by Ag-directed mAb therapy. This study evaluated the activity and safety of a combination of dendritic cell (DC) vaccination given with mAb and cytotoxic therapy. We performed a phase I/II study using autologous DCs pulsed with two different HER2 peptides given with trastuzumab and vinorelbine to a study cohort of patients with HER2-overexpressing and a second with HER2 nonoverexpressing metastatic breast cancer. Seventeen patients with HER2-overexpressing and seven with nonoverexpressing disease were treated. Treatment was well tolerated, with one patient removed from therapy because of toxicity and no deaths. Forty-six percent of patients had stable disease after therapy, with 4% achieving a partial response and no complete responses. Immune responses were generated in the majority of patients but did not correlate with clinical response. However, in one patient, who has survived >14 y since treatment in the trial, a robust immune response was demonstrated, with 25% of her T cells specific to one of the peptides in the vaccine at the peak of her response. These data suggest that autologous DC vaccination when given with anti-HER2-directed mAb therapy and vinorelbine is safe and can induce immune responses, including significant T cell clonal expansion, in a subset of patients.

A prospective cohort study identifies two types of HIV+ Kaposi Sarcoma lesions: proliferative and inflammatory.

Kaposi sarcoma (KS) is the most common cancer in people living with HIV (PLWH) in many countries where KS-associated herpesvirus is endemic. Treatment has changed little in 20 years, but the disease presentation has. This prospective cohort study enrolled 122 human immunodeficiency virus (HIV) positive KS patients between 2017 and 2019 in Malawi. Participants were treated with bleomycin, vincristine and combination antiretroviral therapy, the local standard of care. One-year overall survival was 61%, and progression-free survival was 58%. The 48-week complete response rate was 35%. RNAseq (n = 78) differentiated two types of KS lesions, those with marked endothelial characteristics and those enriched in inflammatory transcripts. This suggests that different KS lesions are in different disease states consistent with the known heterogeneous clinical response to treatment. In contrast to earlier cohorts, the plasma HIV viral load of KS patients in our study was highly variable. A total of 25% of participants had no detectable HIV; all had detectable KSHV viral load. Our study affirms that many KS cases today develop in PLWH with well-controlled HIV infection and that different KS lesions have differing molecular compositions. Further studies are needed to develop predictive biomarkers for this disease.

Mixing rule for calculating the effective refractive index beyond the limit of small particles.

Considering light transport in disordered media, the medium is often treated as an effective medium requiring accurate evaluation of an effective refractive index. Because of its simplicity, the Maxwell-Garnett (MG) mixing rule is widely used, although its restriction to particles much smaller than the wavelength is rarely satisfied. Using 3D finite-difference time-domain simulations, we show that the MG theory indeed fails for large particles. Systematic investigation of size effects reveals that the effective refractive index can be instead approximated by a quadratic polynomial whose coefficients are given by an empirical formula. Hence, a simple mixing rule is derived which clearly outperforms established mixing rules for composite media containing large particles, a common condition in natural disordered media.

Paladin is a phosphoinositide phosphatase regulating endosomal VEGFR2 signalling and angiogenesis.

Cell signalling governs cellular behaviour and is therefore subject to tight spatiotemporal regulation. Signalling output is modulated by specialized cell membranes and vesicles which contain unique combinations of lipids and proteins. The phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2 ), an important component of the plasma membrane as well as other subcellular membranes, is involved in multiple processes, including signalling. However, which enzymes control the turnover of non-plasma membrane PI(4,5)P2 , and their impact on cell signalling and function at the organismal level are unknown. Here, we identify Paladin as a vascular PI(4,5)P2 phosphatase regulating VEGFR2 endosomal signalling and angiogenesis. Paladin is localized to endosomal and Golgi compartments and interacts with vascular endothelial growth factor receptor 2 (VEGFR2) in vitro and in vivo. Loss of Paladin results in increased internalization of VEGFR2, over-activation of extracellular regulated kinase 1/2, and hypersprouting of endothelial cells in the developing retina of mice. These findings suggest that inhibition of Paladin, or other endosomal PI(4,5)P2 phosphatases, could be exploited to modulate VEGFR2 signalling and angiogenesis, when direct and full inhibition of the receptor is undesirable.

The AAA+ ATPase Msp1 is a processive protein translocase with robust unfoldase activity.

Msp1 is a conserved eukaryotic AAA+ ATPase localized to the outer mitochondrial membrane, where it is thought to extract mislocalized tail-anchored proteins. Despite recent in vivo and in vitro studies supporting this function, a mechanistic understanding of how Msp1 extracts its substrates is still lacking. Msp1's ATPase activity depends on its hexameric state, and previous characterizations of the cytosolic AAA+ domain in vitro had proved challenging due to its monomeric nature in the absence of the transmembrane domain. Here, we used a hexamerization scaffold to study the substrate-processing mechanism of the soluble Msp1 motor, the functional homo-hexameric state of which was confirmed by negative-stain electron microscopy. We demonstrate that Msp1 is a robust bidirectional protein translocase that is able to unfold diverse substrates by processive threading through its central pore. This unfoldase activity is inhibited by Pex3, a membrane protein proposed to regulate Msp1 at the peroxisome.

Acute Vasoreactivity Testing and Outcomes in Pulmonary Arterial Hypertension: A Call for Increased Testing.

BACKGROUND: Pulmonary arterial hypertension (PAH) has a progressive, unremitting clinical course. Vasoreactivity testing (VdT) during right heart catheterisation (RHC) identifies a subgroup with excellent long-term response to calcium channel blockade (CCB). Reporting on these patients is limited. Established in 2011, the Pulmonary Hypertension Society of Australia and New Zealand (PHSANZ) registry offers the opportunity to assess the frequency of VdT during RHC, treatment and follow up of PAH patients. METHODS: Registry data from 3,972 PAH patients with index RHC revealed 1,194 VdT appropriate patients. Data was analysed in three groups: 1) VdT+CCB+: VdT positive, CCB treated; 2) VdT+CCB-: VdT positive, no CCB prescribed, 3) VdT-/noVdT: VdT negative, or VdT not tested. Data was reviewed for adherence to guidelines, clinical response (World Health Organization functional class [WHO FC], 6-minute-walk-distance [6MWD], RHC), and outcomes (survival or lung transplantation). RESULTS: Patients included had idiopathic (IPAH=1,087), heritable (HPAH=67) and drug or toxin-induced PAH (DPAH=40). A VdT was performed in 22% (268/1,194), with incomplete data in 26% (70/268); 28% (55/198) were VdT+. Analysis group allocation was: VdT+CCB+ (33/55), VdT+CCB- (22/55), VdT- (143)/noVdT (996). From patients with 1-year data VdT+CCB+ and VdT-/noVdT patients improved WHO FC, 6MWD and cardiac index (CI); VdT+CCB- data remained similar. Within the VdT+CCB+ group, 30% (10/33) were long-term CCB responders with a 100% 5-year survival; non-responders had a 61% survival at 5.4 years. Long-term responders were younger at diagnosis (40 yrs vs 54 yrs). CONCLUSION: Use of VdT testing and documentation is poor in this contemporary patient cohort. Nonetheless, survival in VdT+CCB+ patients from the PHSANZ registry is excellent, supporting guidelines promoting VdT testing. Strategies to promote the use of VdT are warranted.

Localized Antiaromaticity Hotspot Drives Reductive Dehydrogenative Cyclizations in Bis- and Mono-Helicenes.

We describe reductive dehydrogenative cyclizations that form hepta-, nona-, and decacyclic anionic graphene subunits from mono- and bis-helicenes with an embedded five-membered ring. The reaction of bis-helicenes can either proceed to the full double annulation or be interrupted by addition of molecular oxygen at an intermediate stage. The regioselectivity of the interrupted cyclization cascade for bis-helicenes confirms that relief of antiaromaticity is a dominant force for these facile ring closures. Computational analysis reveals the unique role of the preexisting negatively charged cyclopentadienyl moiety in directing the second negative charge at a specific remote location and, thus, creating a localized antiaromatic region. This region is the hotspot that promotes the initial cyclization. Computational studies, including MO analysis, molecular electrostatic potential maps, and NICS(1.7)ZZ calculations, evaluate the interplay of the various effects including charge delocalization, helicene strain release, and antiaromaticity. The role of antiaromaticity relief is further supported by efficient reductive closure of the less strained monohelicenes where the relief of antiaromaticity promotes the cyclization even when the strain is substantially reduced. The latter finding significantly expands the scope of this reductive alternative to the Scholl ring closure.

Phase II Single-Arm Study of Palbociclib and Cetuximab Rechallenge in Patients with KRAS/NRAS/BRAF Wild-Type Colorectal Cancer.

BACKGROUND: Cetuximab is often administered to patients with KRAS wild-type (KRAS-WT) metastatic colorectal cancer (mCRC), although resistance inevitably develops. We hypothesized that co-inhibition of the epidermal growth factor receptor (EGFR) with cetuximab and downstream cyclin-dependent kinases (CDK) 4/6 with palbociclib would be effective for anti-EGFR rechallenge in KRAS-WT mCRC. METHODS: We designed a single-arm, Simon's 2-stage, phase II trial of cetuximab and palbociclib in KRAS-WT mCRC treated with ≥2 prior lines of therapy. We report here on cohort B rechallenging patients with anti-EGFR-based therapy who had disease control of at least 4 months on prior anti-EGFR therapy. Primary endpoint was disease control rate (DCR) at 4 months. RESULTS: Ten evaluable patients were enrolled in this cohort. The 4-month DCR was 20%, which did not fulfill the prespecified 4-month DCR rate to continue. Median progression-free survival was 1.8 months and median overall survival was 6.6 months. Three patients had stable disease, although overall response rate was 0%. Most common treatment-related grades 3-4 adverse events were lymphopenia and leukopenia. CONCLUSION: Selection of patients for anti-EGFR rechallenge using clinical criteria alone was insufficient to identify response to palbociclib + cetuximab. Additional biomarkers are needed to select anti-EGFR rechallenge and circulating tumor DNA (ctDNA) analysis is planned for samples collected in this study. (ClinicalTrials.gov Identifier: NCT03446157).

Brachial plexus anatomy in the miniature swine as compared to human.

Brachial plexus injury (BPI) occurs when the brachial plexus is compressed, stretched, or avulsed. Although rodents are commonly used to study BPI, these models poorly mimic human BPI due to the discrepancy in size. The objective of this study was to compare the brachial plexus between human and Wisconsin Miniature SwineTM (WMSTM ), which are approximately the weight of an average human (68-91 kg), to determine if swine would be a suitable model for studying BPI mechanisms and treatments. To analyze the gross anatomy, WMS brachial plexuses were dissected both anteriorly and posteriorly. For histological analysis, sections from various nerves of human and WMS brachial plexuses were fixed in 2.5% glutaraldehyde, and postfixed with 2% osmium tetroxide. Subsequently paraffin sections were counter-stained with Masson's Trichrome. Gross anatomy revealed that the separation into three trunks and three cords is significantly less developed in the swine than in human. In swine, it takes the form of upper, middle, and lower systems with ventral and dorsal components. Histological evaluation of selected nerves revealed differences in nerve trunk diameters and the number of myelinated axons in the two species. The WMS had significantly fewer myelinated axons than humans in median (p = 0.0049), ulnar (p = 0.0002), and musculocutaneous nerves (p = 0.0454). The higher number of myelinated axons in these nerves for humans is expected because there is a high demand of fine motor and sensory functions in the human hand. Due to the stronger shoulder girdle muscles in WMS, the WMS suprascapular and axillary nerves were larger than in human. Overall, the WMS brachial plexus is similar in size and origin to human making them a very good model to study BPI. Future studies analyzing the effects of BPI in WMS should be conducted.

Fatalistic cancer beliefs and self-reported cancer screening behaviors among diverse urban residents.

Cancer fatalism-the belief that death is inevitable when cancer is present-has been identified as a barrier to cancer screening, detection, and treatment. Our study examined the relationship between self-reported cancer fatalism and adherence to cancer screening guidelines of the breasts, cervix, colon, and prostate among a diverse sample of urban-dwelling adults in Brooklyn, New York. Between May 2019 and August 2020, we conducted a cross-sectional survey of adults 40 + years of age (n = 2,341) residing in Brooklyn neighborhoods with high cancer mortality. Multivariable logistic regression models were used to assess the odds of reporting cancer screening completion across three fatalistic cancer belief categories (low, med, high). Participants' median age was 61 (IQR 51, 71) years, 61% were women, 49% self-identified as non-Hispanic black, 11% Hispanic, 4% Asian, and 6% more than one race. There were no statistically significant differences in the proportion of low, some, or high fatalistic beliefs identified among male respondents compared to women. Among women, we observed that high fatalistic cancer beliefs were associated with higher odds (OR 2.01; 95% CI 1.10-3.65) of completing breast but not cervical (1.04; CI 0.55-1.99) or colon (1.54; CI 0.88-2.69) cancer screening. Men with high fatalistic cancer beliefs had a trend towards lower odds of prostate screening (OR 0.53: 95% CI 0.18-1.57) compared to men with low fatalistic beliefs, but neither was statistically significant. Findings suggest that high fatalistic cancer beliefs may be an important factor in cancer screening utilization among women. Further examination in longitudinal cohorts with a larger sample of men may be needed in order to identify any significant effect.

Comparative retrieval analysis of a novel anatomic tibial tray backside: alterations in tibial component design and surface coating can increase cement adhesions and surface roughness.

BACKGROUND: With the Persona® knee system a novel anatomic total knee design was developed, which has no pre-coating, whereas the predecessor knee system is pre-coated with polymethylmethacrylate (PMMA). Joint registry data have shown no decrease in risk of aseptic revision of PMMA pre-coated tibial components compared with non-pre-coated implants. The aim of this retrieval study was to compare the amount of cement adhesions, geometry and surface features between the two knee designs and to correlate them with the underlying reason for revision surgery. METHODS: Retrieval analysis was performed of 15 NexGen® and 8 Persona® fixed-bearing knee implants from the same manufacturer retrieved from two knee revision centres. A photogrammetric method was used to grade the amount of cement attached to the tibial tray backside. The geometry and dimensions of the tibial trays, tray projections and peripheral lips were measured using digital callipers and compared between the two different designs. To measure the surface roughness on the backside of the tibial tray, a contact profilometer was used. To investigate differences between the two designs statistical analyses (t-test) were performed. RESULTS: All Persona® trays showed evidence of cement adhesion with a % area of 75.4%; half of the NexGen® trays had cement adhesions, with a mean value of 20%. There was a significant difference in the percentage of area covered by cement between the two designs (p < 0.001). Results from the contact profilometer revealed that Persona® and NexGen® tray backsides showed a similar lateral (1.36 µm and 1.10 µm) and medial (1.39 µm and 1.12 µm) mean surface roughness with significant differentiation (p < 0.05) of the lateral and medial roughness values between the two designs. Persona® stems showed a significantly higher mean surface roughness (1.26) compared to NexGen® stems (0.89; p < 0.05). CONCLUSION: The novel anatomic knee system showed significantly more cements adhesions and a higher surface roughness which was most likely attributed to the most obvious design and coating alteration of the tibial tray. This study provides first retrieval findings of a novel TKA design recently introduced to the market.

Correlations of typical pain patterns with SPECT/CT findings in unhappy patients after total knee arthroplasty.

PURPOSE: The diagnostic process in patients after painful total knee arthroplasty (TKA) is challenging. The more clinical and radiological information about a patient with pain after TKA is included in the assessment, the more reliable and sustainable the advice regarding TKA revision can be. The primary aim was to investigate the position of TKA components and evaluate bone tracer uptake (BTU) using pre-revision SPECT/CT and correlate these findings with previously published pain patterns in painful patients after TKA. METHODS: A prospectively collected cohort of 83 painful primary TKA patients was retrospectively evaluated. All patients followed a standardized diagnostic algorithm including 99m-Tc-HDP-SPECT/CT, which led to a diagnosis indicating revision surgery. Pain character, location, dynamics and radiation were systematically assessed as well as TKA component position in 3D-CT. BTU was anatomically localized and quantified using a validated localization scheme. Component positioning and BTU were correlated with pain characteristics using non-parametric Spearman correlations (p < 0.05). RESULTS: Based on Spearman's rho, significant correlations were found between pain and patients characteristics and SPECT/CT findings resulting in nine specific patterns. The most outstanding ones include: Pattern 1: More flexion in the femoral component correlated with tender/splitting pain and patella-related pathologies. Pattern 3: More varus in the femoral component correlated with dull/heavy and tingling/stinging pain during descending stairs, unloading and long sitting in patients with high BMI and unresurfaced patella. Pattern 6: More posterior slope in the tibial component correlated with constant pain. CONCLUSION: The results of this study help to place component positioning in the overall context of the "painful knee arthroplasty" including specific pain patterns. The findings further differentiate the clinical picture of a painful TKA. Knowing these patterns enables a prediction of the cause of the pain to be made as early as possible in the diagnostic process before the state of pain becomes chronic. LEVEL OF EVIDENCE: Level III.

Phase 1 trial of adavosertib (AZD1775) in combination with concurrent radiation and cisplatin for intermediate-risk and high-risk head and neck squamous cell carcinoma.

BACKGROUND: Adavosertib (AZD1775) is an inhibitor of the Wee1 kinase. The authors conducted a phase 1b trial to evaluate the safety of adavosertib in combination with definitive chemoradiotherapy for patients with newly diagnosed, intermediate-risk/high-risk, locally advanced head and neck squamous cell carcinoma (HNSCC). METHODS: Twelve patients with intermediate-risk/high-risk HNSCC were enrolled, including those with p16-negative tumors of the oropharynx, p16-positive tumors of the oropharynx with ≥10 tobacco pack-years, and tumors of the larynx/hypopharynx regardless of p16 status. All patients were treated with an 8-week course of concurrent intensity-modulated radiotherapy at 70 grays (Gy) (2 Gy daily in weeks 1-7), cisplatin 30 mg/m2 weekly (in weeks 1-7), and adavosertib (twice daily on Monday, Tuesday, and Wednesday of weeks 1, 2, 4, 5, 7, and 8). The primary objective was to determine the maximum tolerated dose and the recommended phase 2 dose of adavosertib given concurrently with radiation and cisplatin. Secondary objectives were to determine the 12-week objective response rate and progression-free and overall survival. RESULTS: Three patients (25%) experienced a dose-limiting toxicity, including febrile neutropenia (n = 2) and grade 4 thromboembolism (n = 1). Two dose-limiting toxicities occurred with adavosertib at 150 mg. The median follow-up was 14.7 months. The 12-week posttreatment objective response rate determined by positron emission tomography/computed tomography was 100%. The 1-year progression-free and overall survival rates were both 90%. The maximum tolerated dose of adavosertib was 100 mg. CONCLUSIONS: Adavosertib 100 mg (twice daily on Monday, Tuesday, and Wednesday of weeks 1, 2, 4, 5, 7, and 8), in combination with 70 Gy of intensity-modulated radiotherapy and cisplatin 30 mg/m2 , is the recommended phase 2 dose for patients with HNSCC.

Genomic characteristics and prognostic significance of co-mutated ASXL1/SRSF2 acute myeloid leukemia.

The ASXL1 and SRSF2 mutations in AML are frequently found in patients with preexisting myeloid malignancies and are individually associated with poor outcomes. In this multi-institutional retrospective analysis, we assessed the genetic features and clinical outcomes of 43 patients with ASXL1mut SRSF2mut AML and compared outcomes to patients with either ASXL1 (n = 57) or SRSF2 (n = 70) mutations. Twenty-six (60%) had secondary-AML (s-AML). Variant allele fractions suggested that SRSF2 mutations preceded ASXL1 mutational events. Median overall survival (OS) was 7.0 months (95% CI:3.8,15.3) and was significantly longer in patients with de novo vs s-AML (15.3 vs 6.4 months, respectively; P = .04 on adjusted analysis). Compared to ASXL1mut SRSF2wt and ASXL1wt SRSF2mut , co-mutated patients had a 1.4 and 1.6 times increase in the probability of death, respectively (P = .049), with a trend towards inferior OS (median OS = 7.0 vs 11.5 vs 10.9 months, respectively; P = .10). Multivariable analysis suggests this difference in OS is attributable to the high proportion of s-AML patients in the co-mutated cohort (60% vs 32% and 23%, respectively). Although this study is limited by the retrospective data collection and the relatively small sample size, these data suggest that ASXL1mut SRSF2mut AML is a distinct subgroup of AML frequently associated with s-AML and differs from ASXL1mut SRSF2wt /ASXL1wt SRSF2mut with respect to etiology and leukemogenesis.

Comparative retrieval analysis of antioxidant polyethylene: bonding of vitamin-E does not reduce in-vivo surface damage.

BACKGROUND: With the Persona® knee system a new polyethylene formulation incorporating vitamin-E which aims to reduce oxidation and maintain wear resistance was introduced. Although in-vitro studies have demonstrated positive effects of the vitamin-E antioxidants on UHMWPE, no retrieval study has looked at polyethylene damage of this system yet. It was the aim to investigate the in-vivo performance of this new design, by comparing it with its predecessor in retrieval analysis. METHODS: 15 NexGen® and 8 Persona® fixed-bearing implants from the same manufacturer (Zimmer Biomet) were retrieved from two knee revision centres. For retrieval analysis, a macroscopic analysis of polyethylene using a peer-reviewed damage grading method was used (Hood-score). The roughness of all articulating metal components was measured using a contact profilometer. The reason(s) for TKA revision were recorded. Statistical analyses (t-test) were performed to investigate differences between the two designs. RESULTS: The mean Hood score for Persona® inserts was 109.3 and for NexGen® 115.1 without significant differences between the two designs. Results from the profilometer revealed that Persona® and NexGen® femoral implants showed an identical mean surface roughness of 0.14 µm. The Persona® tibial tray showed a significantly smoother surface (0.06 µm) compared to the NexGen® (0.2 µm; p < 0.001). Both Hood score and surface roughness were influenced by the reasons for revision (p < 0.01). CONCLUSIONS: The bonding of the antioxidant vitamin-E to the PE chain used in the novel Persona® knee system does not reduce in-vivo surface damage compared to highly crosslinked PE without supplemented vitamin-E used in its predecessor knee system NexGen®. However, the Persona® titanium alloy tibial tray showed a significantly smoother surface in comparison to the NexGen® titanium alloy tibial tray. This study provides first retrieval findings of a novel TKA design and may help to understand how the new Persona® anatomic knee system performs in vivo.

A comparison of femoral component rotation after total knee arthroplasty in Kanekasu radiographs, axial CT slices and 3D reconstructed images.

OBJECTIVE: To compare the posterior condylar angle measured with Kanekasu radiograph and 2D-CT with the gold standard 3D-CT following primary total knee arthroplasty (TKA). METHODS: Eighty-two knees with pain following TKA were included in this retrospective study. Two independent raters measured the anatomical and surgical posterior condylar angles twice on each Kanekasu radiograph and 2D-CT. These measurements were compared against the 3D-CT measurement. The intra- and interrater reliability of the Kanekasu radiograph and 2D-CT and the correlation with 3D-CT were calculated. RESULTS: The intra- and interrater reliability for measurements of the anatomical posterior condyle angle for the Kanekasu radiograph and the 2D-CT were excellent for both raters (0.85-0.92). For the less experienced rater 1, the intrarater reliability was significantly better for 2D-CT than Kanekasu radiograph for measuring both the surgical (p < 0.01) and anatomical posterior condyle angles (p < 0.05). For the experienced rater 2, the intrarater reliability was significantly better for Kanekasu radiograph than 2D-CT for measurement of the surgical posterior condyle angle (p < 0.05). The correlation with 3D-CT is higher in 2D-CT than in Kanekasu radiograph (p < 0.01). While the Kanekasu radiograph predicts the 3D-CT angle with 65.9%, 2D-CT can measure the true angle with 82.9% certainty. CONCLUSION: Measurements using the anatomical transepicondylar axis are easier to replicate compared to the surgical transepicondylar axis. In comparison with the gold standard 3D-CT, 2D-CT showed a significantly higher correlation with 3D-CT than the Kanekasu measurements. If 3D-CT is available, it should be preferred over 2D-CT and Kanekasu view radiograph for femoral component rotation measurements.

Typical Pain Patterns in Unhappy Patients After Total Knee Arthroplasty.

BACKGROUND: The primary aim of this study is to assess characteristics of pain in patients with ongoing pain after total knee arthroplasty (TKA). The secondary aim of this study is to identify specific pain patterns and link these to underlying pathologies. METHODS: A prospectively collected cohort of 97 painful primary TKA patients was retrospectively evaluated. All patients followed a standardized diagnostic algorithm, which led to a diagnosis that set the indication for revision surgery. Character, location, dynamics, and radiation of pain were systematically assessed and correlated with the underlying pathologies. RESULTS: Most frequent pain characters were pricking/lancinating (45.7%), pinching/crushing, and dull/heavy (38.6%); 89.5% of all patients localized their knee pain anteriorly; 48.1% reported pain aggravations by descending stairs. Radiating pain was reported in 14% of the patients. Patella-related problems (56.7%) and instability (52.6%) were the most frequent pathologies. Based on correlations between the characteristics 6 specific pain patterns were identified. The most outstanding ones include the following: pattern 1, instability is associated with jumping/shooting, pricking/lancinating and tugging/wrenching pain, and aggravated by chair raising and starting; pattern 6, pain aggravation by descending stairs is associated with anterior and lateral jumping/shooting, tingling/stinging and sharp/lacerating pain character, and TKA positioning and patella baja. CONCLUSION: The assessment of painful TKA patients involving specific pain patterns help to further differentiate and define the clinical picture and ultimately the cause of a painful TKA. If the causes of the described complaints are known, a decision for a therapy can be made reliably and sustainably at an early stage before the state of pain becomes chronic.

Awareness and differential eyeblink conditioning: effects of manipulating auditory CS frequencies.

The role of awareness in differential delay eyeblink conditioning (EBC) has been a topic of much debate. We tested the idea that awareness is required for differential delay EBC when two cues are perceptually similar. The present study manipulated frequencies of auditory conditioned stimuli (CS) to vary CS similarity in three groups of participants. Our findings indicate that awareness was not necessary for differential delay EBC when two tones are easily discriminable, awareness was also not needed for relatively similar tones but may facilitate earlier conditioning, and awareness alone was not sufficient for differential delay EBC.

Changes in Hemodynamic Response Function Resulting From Chronic Alcohol Consumption.

BACKGROUND: Functional MRI (fMRI) task-related analyses rely on an estimate of the brain's hemodynamic response function (HRF) to model the brain's response to events. Although changes in the HRF have been found after acute alcohol administration, the effects of heavy chronic alcohol consumption on the HRF have not been explored, and the potential benefits or pitfalls of estimating each individual's HRF on fMRI analyses of chronic alcohol use disorder (AUD) are not known. METHODS: Participants with AUD and controls (CTL) received structural, functional, and vascular scans. During fMRI, participants were cued to tap their fingers, and averaged responses were extracted from the motor cortex. Curve fitting on these HRFs modeled them as a difference between 2 gamma distributions, and the temporal occurrence of the main peak and undershoot of the HRF was computed from the mean of the first and second gamma distributions, respectively. RESULTS: ANOVA and regression analyses found that the timing of the HRF undershoot increased significantly as a function of total lifetime drinking. Although gray matter volume in the motor cortex decreased with lifetime drinking, this was not sufficient to explain undershoot timing shifts, and vascular factors measured in the motor cortex did not differ among groups. Comparison of random-effects analyses using custom-fitted and canonical HRFs for CTL and AUD groups showed better results throughout the brain for custom-fitted versus canonical HRFs for CTL subjects. For AUD subjects, the same was true except for the basal ganglia. CONCLUSIONS: These findings suggest that excessive alcohol consumption is associated with changes in the HRF undershoot. HRF changes could provide a possible biomarker for the effects of lifetime drinking on brain function. Changes in HRF topography affect fMRI activation measures, and subject-specific HRFs generally improve fMRI activation results.