Comparison of various treatment modalities for the management of bone marrow edema syndrome/transient osteoporosis in men and non-pregnant women: a systematic review.

Transient osteoporosis (TO) or bone marrow edema syndrome (BMES) is a self-limited clinical condition, which affects middle-aged men and women. It can be treated with miscellaneous conservative and surgical measures, which are analyzed in this systematic review. INTRODUCTION: BMES/TO is a transient clinical entity, which can be treated with various therapeutic modalities. The aim of our study was to assess the efficacy of different therapeutic options for the alleviation of pain and reduction of bone marrow edema (BME) in patients with BMES/TO, as well as to propose a therapeutic algorithm. METHODS: PubMed, Scopus, Cochrane, and Google Scholar were searched. Eligibility and extraction of studies were conducted by two authors. Methodological quality assessment was carried out with the modified Delphi technique, Methodological Index for Non-Randomized Studies (MINORS) criteria, and Cochrane Collaboration's risk of bias tool. Outcomes that were compared were time of pain resolution, VAS pain scores, and BME regression on magnetic resonance imaging (MRI). RESULTS: A total of 36 articles (880 patients) were included. Bisphosphonates had higher efficiency in less than 1-month outcomes on pain resolution compared with core decompression (CD), while iloprost was more efficient at 1-3 months compared with bisphosphonates and CD. At 3-6 months, all three of the aforementioned showed equal results on pain resolution, and at a period of 6-12 months, CD and extracorporeal shockwave therapy (ESWT) showed excellent results followed by bisphosphonates and the conservative group (CG) consisting of non-steroidal anti-inflammatory drugs (NSAIDs) and/or analgesics and/or restricted weight bearing. On MRI at 1-3 months, bisphosphonates, iloprost, and CD had relatively the same outcomes on BME resolution, with the least promising being the CG. At 3-6 months, CD seemed to have achieved the best results on the resolution of BME, followed by ESWT, CG, and bisphosphonates group. At 6-12 months, ESWT had the best outcomes compared with the conservative, bisphosphonates, and iloprost groups. CONCLUSION: BMES/TO has been treated with many non-standardized measures due to the low number of highly reliable studies. Current literature shows promising results with regard to the reduction of the clinical course of BMES/TO, but further large multicenter randomized controlled trials, as well as standardized radiological and clinical scores, are warranted to acquire evidence-based recommendations on the therapeutic algorithm.

Potential use of gene drive modified insects against disease vectors, agricultural pests and invasive species poses new challenges for risk assessment.

Potential future application of engineered gene drives (GDs), which bias their own inheritance and can spread genetic modifications in wild target populations, has sparked both enthusiasm and concern. Engineered GDs in insects could potentially be used to address long-standing challenges in control of disease vectors, agricultural pests and invasive species, or help to rescue endangered species, and thus provide important public benefits. However, there are concerns that the deliberate environmental release of GD modified insects may pose different or new harms to animal and human health and the wider environment, and raise novel challenges for risk assessment. Risk assessors, risk managers, developers, potential applicants and other stakeholders at many levels are currently discussing whether there is a need to develop new or additional risk assessment guidance for the environmental release of GD modified organisms, including insects. Developing new or additional guidance that is useful and practical is a challenge, especially at an international level, as risk assessors, risk managers and many other stakeholders have different, often contrasting, opinions and perspectives toward the environmental release of GD modified organisms, and on the adequacy of current risk assessment frameworks for such organisms. Here, we offer recommendations to overcome some of the challenges associated with the potential future development of new or additional risk assessment guidance for GD modified insects and provide considerations on areas where further risk assessment guidance may be required.

Dss1 is a 26S proteasome ubiquitin receptor.

The ubiquitin-proteasome system is the major pathway for protein degradation in eukaryotic cells. Proteins to be degraded are conjugated to ubiquitin chains that act as recognition signals for the 26S proteasome. The proteasome subunits Rpn10 and Rpn13 are known to bind ubiquitin, but genetic and biochemical data suggest the existence of at least one other substrate receptor. Here, we show that the phylogenetically conserved proteasome subunit Dss1 (Sem1) binds ubiquitin chains linked by K63 and K48. Atomic resolution data show that Dss1 is disordered and binds ubiquitin by binding sites characterized by acidic and hydrophobic residues. The complementary binding region in ubiquitin is composed of a hydrophobic patch formed by I13, I44, and L69 flanked by two basic regions. Mutations in the ubiquitin-binding site of Dss1 cause growth defects and accumulation of ubiquitylated proteins.

Diagnostic and therapeutic approach to meniscal ossification: a systematic review.

PURPOSE: The purpose of this study was to systematically review case reports and case series about meniscal ossicle, to summarize existing evidence. Specifically, to identify the etiology, demographic characteristics, localization, clinical features, diagnostic procedures and treatment options of this rare entity. Although, case reports/ series are of low level of evidence, a systematic review of such studies can provide and help us to gain a better understanding and awareness of meniscal ossicle. METHODS: Two authors searched three online databases (MEDLINE, SCOPUS and GOOGLE SCHOLAR) from inception until March 2020 for the literature on meniscal ossicle. Inclusion criteria included case series, case reports and case-based reviews, available in full-text version, in English and that concern humans. Reports published in languages other than English were excluded, as well as articles with no electronic full text availability. Case reports using the term "meniscal ossicle" to describe an acute avulsion fracture of the tibial root of the meniscus, were also excluded. RESULTS: Of 453 initial studies, 38 studies satisfied inclusion criteria. In total 169 patients were included of whom 107 (63%) were males and 62 (37%) were females. Mean age was 44 years (range 12-87). According to Magnetic resonance imaging findings, in 144 knees (86%) the ossicle was localized at the posterior root or horn of the medial meniscus. 60% of the patients had a history of trauma. The predominant symptom in 87% of patients was knee pain. In all patients was detected an intra-articular density structure in computed radiography. 76% had associated meniscal tear, 61% had intraarticular cartilage loss, 34% meniscal extrusion and 28% anterior cruciate ligament injury. Treatment modalities included conservative regimen in 40 patients, while 59 patients underwent surgical excision. CONCLUSION: The most possible etiology of meniscal ossicle is posttraumatic heterotopic ossification and small occult bony avulsion fracture. It is commonly observed in individuals complaining about knee pain with history of antecedent trauma. The presence of a meniscal ossicle should alert the physician to the high likelihood of the patient having an associated meniscal tear, articular cartilage loss, ACL injury or meniscal extrusion. Along with the meniscal ossicle, the associated meniscal tear should be treated as well.

A rare case of polymicrobial chronic maxillary sinusitis due to concurrent isolation of Parvimonas micra and Fusobacterium nucleatum.

Parvimonas micra and Fusobacterium nucleatum are commensal pathogens very rarely isolated simultaneously in clinical specimens. We report a case of chronic maxillary sinusitis caused by these two pathogens, presumably resulting from a co-existing dental infection.

Mps1Mph1 Kinase Phosphorylates Mad3 to Inhibit Cdc20Slp1-APC/C and Maintain Spindle Checkpoint Arrests.

The spindle checkpoint is a mitotic surveillance system which ensures equal segregation of sister chromatids. It delays anaphase onset by inhibiting the action of the E3 ubiquitin ligase known as the anaphase promoting complex or cyclosome (APC/C). Mad3/BubR1 is a key component of the mitotic checkpoint complex (MCC) which binds and inhibits the APC/C early in mitosis. Mps1(Mph1) kinase is critical for checkpoint signalling and MCC-APC/C inhibition, yet few substrates have been identified. Here we identify Mad3 as a substrate of fission yeast Mps1(Mph1) kinase. We map and mutate phosphorylation sites in Mad3, producing mutants that are targeted to kinetochores and assembled into MCC, yet display reduced APC/C binding and are unable to maintain checkpoint arrests. We show biochemically that Mad3 phospho-mimics are potent APC/C inhibitors in vitro, demonstrating that Mad3p modification can directly influence Cdc20(Slp1)-APC/C activity. This genetic dissection of APC/C inhibition demonstrates that Mps1(Mph1) kinase-dependent modifications of Mad3 and Mad2 act in a concerted manner to maintain spindle checkpoint arrests.

Effect of ethanol/TEOS ratios and amount of ammonia on the properties of copper-doped calcium silicate nanoceramics.

Calcium magnesium silicate glasses could be suggested for the synthesis of scaffolds for hard tissue regeneration, as they present a high residual glassy phase, high hardness values and hydroxyapatite-forming ability. The use of trace elements in the human body, such as Cu, could improve the biological performance of such glasses, as Cu is known to play a significant role in angiogenesis. Nano-bioceramics are preferable compared to their micro-scale counterparts, because of their increased surface area, which improves both mechanical properties and apatite-forming ability due to the increased nucleation sites provided, their high diffusion rates, reduced sintering time or temperature, and high mechanical properties. The aim of the present work was the evaluation of the effect of different ratios of Ethanol/TEOS and total amount of the inserted ammonia to the particle size, morphology and bioactive, hemolytic and antibacterial behavior of nanoparticles in the quaternary system SiO2-CaO-MgO-CuO. Different ratios of Ethanol/TEOS and ammonia amount affected the size and morphology of bioactive nanopowders. The optimum materials were synthesized with the highest ethanol/TEOS ratio and ammonia amount as verified by the enhanced apatite-forming ability and antibacterial and non-hemolytic properties.

Color stability of lithium disilicate ceramics after aging and immersion in common beverages.

STATEMENT OF PROBLEM: The color of an esthetic restoration and its color stability are important for long-term success. However, the impact of common beverages on lithium disilicate ceramic is not well known. PURPOSE: The purpose of this in vitro study was to investigate color variations of lithium disilicate ceramics after thermocycling (TC) and immersion in commonly consumed beverages. MATERIAL AND METHODS: A total of 288 specimens (1×10×10 mm) were fabricated from IPS e-max computer-aided design (CAD) (n=72), IPS e-max CERAM ([CER] n=72), IPS e-max Press with glazing ([PG] n=72), and IPS e-max Press without glazing ([PNG] n=72) according to the manufacturer's instructions. Each group was divided into 4 subgroups (n=18): TC, coffee, black tea, and red wine. Thermocycling was performed at 21 900 cycles at 5°C, 37°C, 55°C, and 37°C (3 years' clinical simulation), whereas the specimens were soaked in the staining solutions for up to 54 hours (3 years' clinical simulation). Color parameters L*, a*, and b* were assessed with an ultraviolet-visible spectroscopy recording spectrophotometer. Color difference (ΔE) was calculated using the equation [ΔΕ= [(ΔL*)2 + (Δa*)2 + (Δb*)2]½]. Intraexaminer reliability was assessed by using the intraclass correlation coefficient. Two-way analysis of variance was used for the analysis of ΔΕ, and the parameters L*, a*, and b* were analyzed with linear mixed models for repeated measurements and the Bonferroni pair-wise comparison test (α=.05). RESULTS: Parameters ΔΕ, L*, a*, and b* were significantly affected by the interaction between material and treatment (P<.001). A ΔΕ >1 was recorded for PG with tea, wine, and coffee, PG after TC and CER after TC. For L*, greater reduction was observed for PNG with tea and CER after TC, whereas for a* significant changes were positive (to red shades), except for PNG with TC, where PNG with wine showed the greatest positive change. For b*, significant changes were negative (to blue shades) except for PNG with tea and coffee and CAD with tea. CONCLUSIONS: All groups demonstrated color changes below the clinically perceptible level (ΔΕ<3.7), except PNG in tea which showed statistically significant color differences (ΔΕ>4). CAD presented higher color stability compared with the nonglazed Press specimens.

Odontogenic differentiation and biomineralization potential of dental pulp stem cells inside Mg-based bioceramic scaffolds under low-level laser treatment.

This study aimed to investigate the potential of low-level laser irradiation (LLLI) to promote odontogenic differentiation and biomineralization by dental pulp stem cells (DPSCs) seeded inside bioceramic scaffolds. Mg-based, Zn-doped bioceramic scaffolds, synthesized by the sol-gel technique, were spotted with DPSCs and exposed to LLLI at 660 nm with maximum output power of 140 mw at fluencies (a) 2 and 4 J/cm2 to evaluate cell viability/proliferation by the MTT assay and (b) 4 J/cm2 to evaluate cell differentiation, using real-time PCR (expression of odontogenic markers) and a p-nitrophenylphosphate (pNPP)-based assay for alkaline phosphatase (ALP) activity measurement. Scanning electron microscopy (SEM) and X-ray diffraction (XRD) analysis were used for structural/chemical characterization of the regenerated tissues. Exposure of the DPSCs/scaffold complexes to the proposed LLLI scheme was associated with statistically significant increase of odontogenesis-related markers (bone morphogenetic protein 2 (BMP-2): 22.4-fold, dentin sialophosphoprotein (DSPP): 28.4-fold, Osterix: 18.5-fold, and Runt-related transcription factor 2 (Runx2): 3.4-fold). ALP activity was significantly increased at 3 and 7 days inside the irradiated compared to that in the non-irradiated SC/DPSC complexes, but gradually decreased until 14 days. Newly formed Ca-P tissue was formed on the SC/DPSC complexes after 28 days of culture that attained the characteristics of bioapatite. Overall, LLLI treatment proved to be beneficial for odontogenic differentiation and biomineralization of DPSCs inside the bioceramic scaffolds, making this therapeutic modality promising for targeted dentin engineering.

Is There an Association between a Tonsillar Diffuse Large B-Cell Lymphoma Arising after a Neck Squamous Cell Carcinoma of Occult Primary? A Case Report and Extensive Literature Review.

OBJECTIVES: The aim of this review is to focus on the possibility of patients with squamous cell carcinoma to develop a second primary disease such as DLBCL, perhaps because of the irradiation of the head and neck area. MATERIALS AND METHODS: A case of an 89-year-old man is reported, who initially underwent surgical and complementary treatment for neck squamous cell carcinoma of occult primary and later for tonsillar diffuse large B-cell non-Hodgkin lymphoma. RESULTS: The second primary was considered a recurrence in the neck of the original cancer of unknown primary, so a new surgical management was decided. The final pathology report described a diffuse large B-cell non-Hodgkin lymphoma. CONCLUSIONS: The importance of maintaining follow-ups for patients with occult primary cancers who are at an elevated risk of developing a metastasis or a second primary carcinoma outbreak is highlighted.

The Effect of the COVID-19 Pandemic on Odontogenic Cervicofacial Infections in a Single Center in Greece.

INTRODUCTION: Odontogenic cervicofacial infections are still an ongoing problem, requiring immediate hospital admittance and management. The aim of this study is to reflect the number of patients with cervicofacial infections who were admitted during the coronavirus disease 2019 (COVID-19) pandemic period in a single, point of reference center in Northern Greece as well as analyze the quantitative and qualitative parameters of patient characteristics and management data. METHODS: This was a retrospective cohort study that included all the patients with cervicofacial infections who were admitted to our unit during the COVID-19 pandemic, specifically between 2020 and 2021. For comparative reasons, patients admitted with cervicofacial infections between 2019 and 2020 (pre-COVID period) were analyzed. RESULTS: In total, 341 patients fulfilled the criteria for this study. Specifically, the number of admitted patients was 151 in the pre-COVID era instead of 190 patients in the pandemic. The mean age of the patients was 45.3 years, with a slight male predominance (54.7% males to 45.3%). The mean duration of hospitalization was 2.5 days in the pre-COVID period instead of 3.42 days in the pandemic. Interestingly, in the pandemic, eight times more patients were admitted to the ICU post-operatively, in contrast to the pre-COVID period (23 vs 3 patients). Also in the COVID period, almost 54.9% of the patients presented with fever and 49.6% with trismus. Moreover, the submandibular space involvement was the most common space of infection in both COVID and pre-COVID groups with (58.9% and 49.7%) respectively. In one-third of all cases, a post-extraction infection of a third molar was the main cause of abscess. CONCLUSION: Cervicofacial infections during the COVID-19 pandemic appeared with more severe symptoms and resulted in an increased number of patients who needed admittance to the intensive care unit, in contrast to the pre-COVID era. Also, the mean length of stay was increased for a day at the same period. This study could be used as an example for further research, in case of similar pandemic situations in the future.

Corneal changes after large (9mm) lateral rectus muscle recession measured with Pentacam®.

Introduction: The aim of this study was to evaluate alterations in corneal astigmatism, axial anterior corneal curvature, anterior chamber depth, and central corneal thickness (CCT) two months after the unilateral recession of lateral rectus muscle in children. Methods: This prospective study included 37 children with intermittent exotropia who would undergo unilateral lateral rectus muscle recession. All measurements were performed using Pentacam®. Comparisons were made between the operated and fellow unoperated eyes, pre- and post-operatively. The assessment was made for changes in the radius of axial curvature on major meridians at 3 and 3.5 mm from the optical corneal center in the mid-peripheral zone. Astigmatism changes of the anterior and posterior corneal surface were calculated using vector analysis software (astigMATIC®). The interaction between age or CCT and postoperative changes in anterior and posterior surface corneal astigmatism were examined with ANOVA model. Results: In the intervention group, changes in anterior and posterior corneal surface astigmatism were statistically significant, with a mean increase of 0.56Dx90 and 0.08Dx87, respectively. In the mid-peripheral corneal zone, an increase was observed in the radius of anterior corneal axial curvature, more evident temporal 3 and 3.5 mm from the corneal center on the horizontal meridian, with corresponding decrease superiorly and inferiorly at 3 and 3.5 mm from the corneal center on the vertical meridian. Discussion: The changes in total astigmatism of the operated eyes are mainly attributed to the anterior corneal surface. These changes are associated with flattening in the 180 meridian of the cornea, leading to a shift to "with-the-rule" astigmatism.

Graft-Versus-Host Disease: Can Biomarkers Assist in Differential Diagnosis, Prognosis, and Therapeutic Strategy?

A crucial complication after allogeneic hematopoietic cell transplantation (alloHCT), namely, acute graft-versus-host disease (aGVHD), occurs in about 50% of transplant recipients, leading to high morbidity and mortality. Thus far, the diagnosis of GVHD has been mainly established through clinical features and histologic or laboratory evidence of periductal lymphocyte infiltration, fibroplasia, and mixed lymphocytic and plasmocytic inflammation. Intensive research is focused on identifying biomarkers for the early diagnosis, prediction of disease, response to treatment, prognosis, and risk stratification of patients. The serum biomolecules that have been investigated are reported and summarized. Moreover, oral tissue involvement in GVHD is described, and other biomarkers that have been proposed, such as saliva, are analyzed. Future research is highlighted as a necessity in order for these biomarkers to be validated and quantified for use in clinical practice.

A Preliminary Study of the Role of Endothelial-Mesenchymal Transitory Factor SOX 2 and CD147 in the Microvascularization of Oral Squamous Cell Carcinoma.

INTRODUCTION: The aim of this study was to detect the possible endothelial expression of embryonic-type cancer stem cells (CSC) marker SOX2 and the stemness-type CSC marker CD147 in oral potential malignant disorders (OPMDs), oral leukoplakia (OL) in particular, and oral squamous cell carcinoma (OSCC). METHODS: This study focuses on the immunohistochemical pattern of expression of CSC protein-biomarkers SOX2 and CD147 in paraffin-embedded samples of 21 OSCCs of different grades of differentiation and 30 cases of OLs with different grades of dysplasia, compared to normal oral mucosa. RESULTS: The protein biomarker SOX2 was expressed in the endothelial cells, but without establishing any statistically significant correlation among OSCC, OL, and normal tissue specimens. However, SOX endothelial staining was noticed in 7/30 (23.3%) cases of OL (one non-dysplastic, one mildly dysplastic, one moderately dysplastic, and four severely dysplastic cases) and 5/21 (23.8%) cases of OSCC (two well-differentiated, one moderately differentiated, and two poorly differentiated cases). Although CD147 is expressed in normal oral epithelium, OL, and OSCC neoplastic cells, its vascular-endothelial expression was noticed in only 2/5 (40%) cases of normal oral epithelium, 1/30 (3.3%) cases of OL (one severely dysplastic case), and 4/21 (19%) cases of OSCC (two well-differentiated, one moderately differentiated, and one poorly differentiated case). Therefore, no statistically significant correlation among OSCC, OL, and normal tissue specimens was established. CONCLUSION: The endothelial presence of SOX2 both in oral potentially malignant and malignant lesions suggests that SOX2 may be implicated in the microvascularization process and associated with the degree of dysplasia in OL. The expression of CD147 may be attributed both to local inflammation and tumorigenesis. The implementation of CD147 in larger groups of tissue samples will shed some light on its role in cancer and inflammation. The evidence so far supports the need for more studies, which may support the clinical significance of these novel cancer stem cell biomarkers.

Corneal tomography changes following major (6 mm) medial rectus muscle recession: a prospective cohort study using Pentacam®.

INTRODUCTION: The aim of this study is to evaluate changes in corneal astigmatism, axial anterior corneal curvature, as well as changes in the anterior chamber depth and central corneal thickness, 2 months following the unilateral recession of medial rectus muscle in children. METHODS: Thirty-three children with esotropia were prospectively evaluated following unilateral medial rectus muscle recession, using Pentacam®. Comparisons were made between the operated and fellow unoperated eyes, pre, and postoperatively. The assessment was made for changes in the radius of axial curvature on major meridians at 3 and 3.5 mm from the optical corneal center in the mid-peripheral zone. Astigmatism changes of the anterior and posterior corneal surface were calculated using vector analysis software (astigMATIC®). ANOVA model was used to examine the interaction between age or central corneal thickness and postoperative changes in anterior and posterior surface corneal astigmatism. RESULTS: In the intervention group, changes in anterior and posterior corneal surface astigmatism were statistically significant, with a mean increase of 0.59Dx92 and 0.08Dx91, respectively. In the mid-peripheral corneal zone, there is an increase in the radius of anterior corneal axial curvature more evident nasally 3.5 mm from the corneal center on the horizontal meridian, with corresponding decrease superiorly and inferiorly at 3 and 3.5 mm from the corneal center on the vertical meridian. DISCUSSION: The changes in total astigmatism of the operated eyes are mainly attributed to the anterior corneal surface. These changes are associated with flattening in the 180 meridian of the cornea, leading to a shift to "with-the-rule" astigmatism.

Sentinel lymph node biopsy versus elective neck dissection in management of the clinically negative (cN0) neck in patients with oral squamous cell carcinoma: A systematic review and meta-analysis.

Oral squamous cell carcinoma (OSCC) is the most prevalent type of head and neck cancer, and lymph node metastasis is a crucial prognostic factor that has a direct correlation with the survival rate. The standard procedure for managing clinically negative (cN0) neck in OSCC patients is elective neck dissection (END), but it can lead to various complications that affect the patient's quality of life. Therefore, medical professionals are exploring the use of the sentinel lymph node biopsy (SLNB) to manage cN0 neck in OSCC patients. The aim of this systematic review and meta-analysis was to compare the overall survival (OS) and disease-free survival (DFS) of patients undergoing SLNB and END in the surgical management of early cN0 neck in OSCC patients. To conduct this study, the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement was used to report the systematic reviews. The study included all comparative clinical studies, including randomized clinical trials (RCTs), clinical perspective studies, and retrospective studies that compared END to SLNB for early cN0 neck in OSCC patients. The hazard ratio (HR) and risk ratio (RR) with 95% confidence interval (CI) were calculated using comprehensive meta-analysis. The primary outcome variables were OS, DFS, and disease-specific survival (DSS). The secondary outcome variable was the nodal recurrence rate without local or distant metastasis. The GRADE system was used to assess the evidence's certainty. The meta-analysis included 12 clinical studies, comprising three RCTs and seven non-RCTs. The results showed no statistically significant difference between END and SLNB concerning OS (HR = 0.993, CI: 0.814 to 1.211, P = 0.947, low-quality evidence), DFS (HR = 0.705, CI: 0.408 to 1.29, P = 0.21, low-quality evidence), and nodal recurrence (RR = 1.028, CI: 0.969 to 1.090, P = 0.907, low-quality evidence). Additionally, SLNB was non-inferior to END regarding DSS (RR = 1.028, CI: 0.969 to 1.090, P = 0.907, low-quality evidence). In conclusion, this research affirms the safety and precision of SLNB as a means to stage cN0 OSCC patients, with outcomes that are comparable to END in terms of survival and nodal recurrence. Nevertheless, it's crucial to acknowledge that the quality of evidence in this study was relatively low. Therefore, additional RCTs comparing END to SLNB are warranted to validate these results and offer more robust guidance for managing cN0 OSCC patients.

The ubiquitin-associated (UBA) 1 domain of Schizosaccharomyces pombe Rhp23 is essential for the recognition of ubiquitin-proteasome system substrates both in vitro and in vivo.

The ubiquitin-proteasome system is essential for maintaining a functional cell. Not only does it remove incorrectly folded proteins, it also regulates protein levels to ensure their appropriate spatial and temporal distribution. Proteins marked for degradation by the addition of Lys(48)-linked ubiquitin (Ub) chains are recognized by shuttle factors and transported to the 26 S proteasome. One of these shuttle factors, Schizosaccharomyces pombe Rhp23, has an unusual domain architecture. It comprises an N-terminal ubiquitin-like domain that can recognize the proteasome followed by two ubiquitin-associated (UBA) domains, termed UBA1 and UBA2, which can bind Ub. This architecture is conserved up to humans, suggesting that both domains are important for Rhp23 function. Such an extent of conservation raises the question as to why, in contrast to all other shuttle proteins, does Rhp23 require two UBA domains? We performed in vitro Ub binding assays using domain swap chimeric proteins and mutated domains in isolation as well as in the context of the full-length protein to reveal that the Ub binding properties of the UBA domains are context-dependent. In vivo, the internal Rhp23 UBA1 domain provides sufficient Ub recognition for the protein to function without UBA2.

Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation.

The ubiquitin-proteasome system targets selected proteins for degradation by the 26S proteasome. Rpn12 is an essential component of the 19S regulatory particle and plays a role in recruiting the extrinsic ubiquitin receptor Rpn10. In the present paper we report the crystal structure of Rpn12, a proteasomal PCI-domain-containing protein. The structure helps to define a core structural motif for the PCI domain and identifies potential sites through which Rpn12 might form protein-protein interactions. We demonstrate that mutating residues at one of these sites impairs Rpn12 binding to Rpn10 in vitro and reduces Rpn10 incorporation into proteasomes in vivo.