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Unifying terminology used to describe morphologic features is a very important endeavour to assure comparability of work and papers on osteoarthritis animal models. In this editorial an attempt is presented to define and unify the terminology of the macroscopic and histological description of joint changes in OA for both human OA and the OA animal models. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented in response to the article about the histologically distinct picture of pancreatitis-type fat necrosis in a resection specimen from a female patient.

Abstract Epstein-Barr virus (EBV)-associated B-cell post-transplantation lymphoproliferative disorder (PTLD) is a severe complication following stem cell transplantation. This is believed to occur as a result of iatrogenic immunosuppression leading to a relaxation of T-cell control of EBV infection and thus allowing viral reactivation and proliferation of EBV-infected B-lymphocytes. In support of this notion, reduction of immunosuppressive therapy may lead to regression of PTLD. We present a case of an 18-year-old male developing a monomorphic B-cell PTLD 2 months after receiving an allogenic stem cell transplant for acute lymphoblastic leukemia. Reduction of immunosuppressive therapy led to regression of lymphadenopathy. Nevertheless, the patient died 3 months afterwards due to extensive graft-vs.-host-disease and sepsis. As a diagnostic lymph node biopsy was performed only after reduction of immunosuppressive therapy, we are able to study the histopathological changes characterizing PTLD regression. We observed extensive apoptosis of blast cells, accompanied by an abundant infiltrate comprising predominantly CD8-positive, Granzyme B-positive T-cells. This observation supports the idea that regression of PTLD is mediated by cytotoxic T-cells and is in keeping with the observation that T-cell depletion, represents a major risk factor for the development of PTLD.

Abstract Primary liver tumors in children are rare with malignant hepatoblastoma being the most common neoplasm. In this report, we describe the diagnosis and clinical management of a large liver tumor in a 3-year-old child that displayed the features of both, conventional hepatoblastoma and malignant teratoma. Pathological assessment on a pre-operative bioptical specimen showed an immature teratoid tumor with no area of hepatoblastic differentiation present. Histological and immunhistological examination of the resected tumor specimen additionally showed tumor areas of very different differentiation pattern intermixed with each other, namely areas of hepatoblastoma-typical and neuroblastoma-like morphology as well as areas of rhadomyosarcomatous differentiation. After chemotherapy the tumor size increased and an extended right hemihepatectomy was performed. Post-operatively, the general condition of the child improved and adjuvant chemotherapy was started two weeks later. 36 months after initial diagnosis the patient is healthy, in good general condition, and without any sign of residual tumor disease. Overall, we describe the diagnosis and clinical management of a large liver tumor in a 3-year-old child that displayed the features of both, conventional hepatoblastoma and malignant teratoma and was designated as mixed hepatoblastoma and teratoma. Though mesenchymal tumor portions can occur within hepatoblastomas, most commonly osteoid or chondroid, our case is different as it presents a large spectrum of mesenchymal and epithelial differentiation pattern in most of the lesion.

SUMMARY: Histological and histochemical methods are important tools in the evaluation of joint tissue samples for degenerative joint diseases, both in humans and in animal models. In this respect, standardized, simple, and reliable techniques are mandatory. This chapter describes five basic staining procedures appropriate for macroscopic (Indian ink) and histologic (HE/hematoxylin - eosin) visualization and scoring of cartilage proteoglycan and collagen content (toluidine blue/safranin O and picrosirius red/Goldner''s trichrome). [Copyright &y& Elsevier]

Summary: Animal model systems represent an important adjunct and surrogate for studies of osteoarthritis (OA) in humans. They provide a means to study OA pathophysiology as well as aid in the development of therapeutic agents and biological markers for diagnosing and prognosing the disease. Thus, it is of great importance for the OA scientific community, both in academic as well as industrial research, to standardize scoring systems for evaluating the OA disease process and to make results between different studies comparable. The task of the histopathology initiative of OARSI was to achieve a consensus of scoring systems for the most important species used in OA animal model research (dog, guinea pig, horse, mouse, rabbit, rat, and sheep/goat), which are presented in the various chapters in this special volume of Osteoarthritis & Cartilage together with extra chapters on basic methodology (histochemistry, statistics, morphometry), the specific terminology and a general discussion of animal models in OA research. Standardized definitions are suggested for basic but essential terms such as “grading” and “staging” in order to promote their consistent use and thereby promote improved understanding and data interpretation across all model systems. Thus, this introductory chapter presents an overview of the guiding principles for assessment of important OA animal model systems. Use of such systems, independently or in conjunction with other systems in parallel, should facilitate comparability of results across animal model studies. [ABSTRACT FROM AUTHOR]

Objective: Diabetic glomerulosclerosis is characterized by thickening of the glomerular basement membrane and diffuse or nodular expansion of the mesangial matrix. The microfibrillar collagen type VI is one of the extracellular matrix compounds that is suggested to account for mesangial matrix expansion although the current data is contradictory. Therefore, it was the aim of the present study to analyse the glomerular expression of collagen VI in human biopsies with diabetic and hypertensive nephropathy as well as in experimental models of type 1 and type 2 diabetic nephropathy. Methods: Expression and distribution of collagen VI and collagen IV was investigated in experimental models of type 1 (Streptozotocin, STZ) and 2 (SHR-Nc/p) diabetic nephropathy (n = 5 per group) and control groups (sham and subtotally nephrectomized rats, SNX, n = 6 per group) using immunohistochemistry and a semiquantitative scoring system. In addition, kidney biopsies from patients with diabetic nephropathy (DN) and hypertensive-diabetic nephropathy (HDN) were studied. Patient age ranged from 40-70 years (mean: 63). As controls renal tissue from 6 age and sex matched healthy subjects was included in the study. Results: In normal human and rat kidney, collagen VI staining was predominantly localized to the tubulointerstitium whereas no or only very weak expression was found in the glomerulus. In human DN and in HDN as well as in both models of experimental diabetes interstitial collagen VI expression was markedly and comparably upregulated. In contrast, glomerular collagen VI expression score was significantly higher in DN (3.2) compared to HDN (2.0) and controls (0.4) and in both models of diabetic nephropathy (STZ: 1.64 ± 0.75, SHR/N-cp: 1.04 ± 0.46) compared to controls (0.70 ± 0.30) and to SNX (0.74 ± 0.38). In DN with nodular glomerulosclerosis there was a specific expression pattern of collagen VI with increased staining in the basement membrane around the nodules. Collagen IV expression was absent or low in normal glomeruli and was upregulated in DN and HDN as well as in both experimental diabetic diseases and in SNX. Of note, glomerular collagen IV expression was highest in areas of segmental sclerosis. Conclusions: Renal interstitial and in particular glomerular collagen VI protein expression was significantly upregulated in experimental models of type 1 and 2 diabetes mellitus as well as in human diabetic nephropathy. The data and the expression pattern of collagen VI support a specific upregulation of the microfbrillar collagen type VI in diffuse diabetic glomerulosclerosis. [ABSTRACT FROM AUTHOR]

Objective: BMP-7 plays an important role during fetal kidney development and is expressed multifocally within the metanephric mesenchyme. BMP-7 knockout mice die due to renal failure perinatally as result of severe kidney dysplasia. In the adult, BMP-7 is most strongly expressed in the kidney. In addition, there is recent experimental evidence that BMP-7 protects tubular epithelial cells from ischemic damage and prevents or reverses interstitial fibrosis. However, the exact expression pattern as well as the function of BMP-7 in the adult kidney are unclear. Methods: Normal adult kidney samples were collected immediately after tumor surgery from non-affected kidney portions. Immunostainings were performed for BMP-7, Phospho-Smad 1,5,8 (as mediators of BMP-7 activities) as well as for calbindin (as a marker of distal nephron) and for uromucoid (i.e. Tamm Horsefall protein as a marker for the thick ascending limb) using standard immunofluorescence technologies. Results: BMP-7 expression was localized specifically to the epithelia of the distale tubulus as well as the collecting ducts as identified by co-immunostaining with calbindin. Of interest, prominent immunostaining was found in the luminal cell compartment. No expression was detected in the proximal tubulus and the glomeruli. Phospho-Smad1,5,8 expression colocalized with BMP-7 and was also restricted to the epithelia of the distale tubulus and the collecting ducts. Conclusions: Our data indicate that BMP-7 expression in the adult kidney is much more restricted than in the fetal situation and selectively found in the distale nephron. Also cellular reactivity to BMP-stimulation i.e. Phospho-Smad expression was found there. Altogether, the findings suggests that BMP-7 plays a physiological role only in this part of the kidney. Still, as reported previously, proximale tubular cells might be responsive to BMP-7, but not in an autocrine or paracrine mode, but maybe only if externally given. The luminal distribution of the immunostaining most likely implicates urinary excretion of BMP-7 in line with distal signalling activity. [ABSTRACT FROM AUTHOR]

Objective: There is increasing evidence that atherosclerosis is more common and more severe in patient with renal failure compared to the non renal population. Since atherosclerosis in renal failure is characterized by increased media and plaque calcification it was suggested that factors usually involved in enchondral ossification may also be expressed in the vasculature. Therefore, it was the aim of the present study to analyse the expression of some cartilage-specific markers in atherosclerotic lesions of uremic and non-uremic patients. Methods: Atherosclerotic lesions in autoptic coronary specimens of 24 patients with or without chronic renal failure were classified from type II to type VIII lesion according to Stary on HE and Kossa stains. Thereafter, using immunhistochemistry and a semiquantitative scoring system the specimens were analysed for in-situ protein expression of the following mineralizing regulating, cartilage specific proteins: aggregating chondroitin sulphate proteoglycan (aggrecan), S100, collagen type I and type X. Results: Heavy media calcification was seen in 12 out of the 24 specimens. Qualitative analysis of the coronary arteries showed significantly more calcified plaques of coronary arteries in patients with end-stage renal failure than in non-renal patients. Protein expression of aggrecan was significantly higher in the media beyond atherosclerotic plaques. Inside the plaques, aggrecan protein was also expressed by vascular smooth muscle cells. The expression pattern differed between the plaque subregions, and was particularly high in the regions of the plaque where calcification was seen (i.e. Stary VII lesion). Furthermore, the expression was enhanced in advanced atherosclerotic lesions from patients with chronic failure compared to non-renal controls. The non calcified human arterial samples did not stain positively for Aggrecan. The coronary specimen studied did not expressed S100 protein, collagen type I or collagen type X, respectively. Conclusions: The data provide no evidence for a major role of cartilage-specific mineralization regulating proteins, i.e. enchondral ossification, in advanced atherosclerosis in renal failure. However, the study provides circumstantial evidence for a role of aggrecan, a major water binding protein, for media degeneration and subsequent calcification. [ABSTRACT FROM AUTHOR]

Bone morphogenetic protein-7 (BMP)-7 plays an important role during fetal kidney development. In the adult, BMP-7 is most strongly expressed in the kidney compared to other organs, but the exact expression pattern as well as the function of BMP-7 is unclear. The major aim of the present study was to define which parts of the human kidney do physiologically express BMP-7 and which cells appear to be targets of BMP activity by showing phosphorylated BMP-receptor-associated Smads 1, 5, or 8 and inhibitor of differentiation factor 1 (ID1) expression. BMP-7 expression was localized by immunohistology to the epithelia of the distal tubule as well as the collecting ducts (CDs). Phospho-Smads 1/5/8 and ID1 expression largely colocalized with BMP-7 and was also localized in the epithelia of the distal tubule and the CDs. This was confirmed by polymerase chain reaction-based mRNA expression analysis. In vitro, proximal tubular cells (PTCs) expressed BMP receptors and BMP-receptor-associated Smads and were reactive to BMP-7. Our data indicate that BMP-7 expression in the adult human kidney appears to be more restricted than in the fetal situation and predominantly found in the distal nephron. Also, evidence of in vivo BMP signalling (i.e. phospho-Smads and ID1 expression) was found there. These findings suggest that BMP-7 plays a physiological role mostly in this part of the kidney. Still, as reported previously, PTCs are responsive to BMP-7, but presumably not in an autocrine or paracrine mode in normal adult kidneys. [ABSTRACT FROM AUTHOR]

The extracellular matrix represents a complex alloy of variable members of diverse protein families defining structural integrity and various physiological functions. The most abundant family is the collagens with more than 20 different collagen types identified so far. Collagens are centrally involved in the formation of fibrillar and microfibrillar networks of the extracellular matrix, basement membranes as well as other structures of the extracellular matrix. This review focuses on the distribution and function of various collagen types in different tissues. It introduces their basic structural subunits and points out major steps in the biosynthesis and supramolecular processing of fibrillar collagens as prototypical members of this protein family. A final outlook indicates the importance of different collagen types not only for the understanding of collagen-related diseases, but also as a basis for the therapeutical use of members of this protein family discussed in other chapters of this issue. [Copyright &y& Elsevier]

Objective. To investigate the expression and target genes of pigment epithelium-derived factor (PEDF) in cartilage and chondrocytes, respectively. Methods. We analyzed the expression pattern of PEDF in different human cartilaginous tissues including articular cartilage, osteophytic cartilage, and fetal epiphyseal and growth plate cartilage, by immunohistochemistry and quantitative real-time (qRT) PCR. Transcriptome analysis after stimulation of human articular chondrocytes with rhPEDF was performed by RNA sequencing (RNA-Seq) and confirmed by qRT-PCR. Results. Immunohistochemically, PEDF could be detected in transient cartilaginous tissue that is prone to undergo endochondral ossification, including epiphyseal cartilage, growth plate cartilage, and osteophytic cartilage. In contrast, PEDF was hardly detected in healthy articular cartilage and in the superficial zone of epiphyses, regions that are characterized by a permanent stable chondrocyte phenotype. RNA-Seq analysis and qRT-PCR demonstrated that rhPEDF significantly induced the expression of a number of matrix-degrading factors including SAA1, MMP1, MMP3, and MMP13. Simultaneously, a number of cartilage-specific genes including COL2A1, COL9A2, COMP, and LECT were among the most significantly downregulated genes. Conclusions. PEDF represents a marker for transient cartilage during all neonatal and postnatal developmental stages and promotes the termination of cartilage tissue by upregulation of matrix-degrading factors and downregulation of cartilage-specific genes. These data provide the basis for novel strategies to stabilize the phenotype of articular cartilage and prevent its degradation. [ABSTRACT FROM AUTHOR]

Abstract: Anabolic and catabolic cytokines and growth factors such as BMP-7 and IL-1β play a central role in controlling the balance between degradation and repair of normal and (osteo)arthritic articular cartilage matrix. In this report, we investigated the response of articular chondrocytes to these factors IL-1β and BMP-7 in terms of changes in gene expression levels. Large scale analysis was performed on primary human adult articular chondrocytes isolated from two human, independent donors cultured in alginate beads (non-stimulated and stimulated with IL-1β and BMP-7 for 48h) using Affymetrix gene chips (oligo-arrays). Biostatistical and bioinformatic evaluation of gene expression pattern was performed using the Resolver software (Rosetta). Part of the results were confirmed using real-time PCR. IL-1β modulated significantly 909 out of 3459 genes detectable, whereas BMP-7 influenced only 36 out of 3440. BMP-7 induced mainly anabolic activation of chondrocytes including classical target genes such as collagen type II and aggrecan, while IL-1β, both, significantly modulated the gene expression levels of numerous genes; namely, IL-1β down-regulated the expression of anabolic genes and induced catabolic genes and mediators. Our data indicate that BMP-7 has only a limited effect on differentiated cells, whereas IL-1β causes a dramatic change in gene expression pattern, i.e. induced or repressed much more genes. This presumably reflects the fact that BMP-7 signaling is effected via one pathway only (i.e. Smad-pathway) whereas IL-1β is able to signal via a broad variety of intracellular signaling cascades involving the JNK, p38, NFkB and Erk pathways and even influencing BMP signaling. [Copyright &y& Elsevier]

Abstract: Sequence stratigraphy in deeper water, epicontinental carbonates such as in the Upper Jurassic of southern Germany is difficult because the recognition of parasequences, sequences and sequence boundaries is impeded by the paucity in diagnostic sedimentological criteria or stratal surfaces. Using the “genetic stratigraphic” approach, and integrating facies, stable isotope (C, O) and palynofacies analysis two types of genetic depositional sequences can be discriminated: small-scale sequences are stacked into medium-scale sequences which may record a 400 kyr Milankovitch signal. The medium-scale sequences were correlated regionally using both gamma-ray logs and stable isotope records. Regional correlations show that the depocentres are controlled by underlying palaeotectonic elements (Late Palaeozoic troughs). The rise/fall turnarounds of medium-scale sequences are marked by negative δ18O peaks (temperature maxima) and reduced absolute palynoclast contents. The fall/rise turnarounds are marked by positive δ18O peaks (temperature minima) and high absolute palynoclast contents. The initiation and termination of sponge/microbial mounds show characteristic patterns: thrombolitic microbialites form during intervals of (1) reduced input of terrestrial palynomorphs interpreted as an increase in distality, (2) decreasing δ13C trends interpreted to be related to decreasing nutrient supply and (3) decreasing δ18O values interpreted as phases of warming and rising relative sea-level. In contrast, thrombolitic/stromatolitic microbialites were found to occur during phases of (1) increasing input of terrestrial palynomorphs interpreted as an increase in proximality, (2) increasing δ13C values interpreted to reflect increasing terrestrial input and nutrient supply as well as increasing δ18O values (interpreted as phases of cooling and relative sea-level falls). Isotopic and palynofacies evidence suggests that bioherms were terminated by sudden input of nutrients during relative sea-level falls. Sedimentological criteria were clearly not sufficient to delineate a robust sequence stratigraphic framework. [Copyright &y& Elsevier]

This article presents the rare case of a 54-year-old gentleman with primary glioblastoma developing multiple extracranial metastases 7 months after diagnosis. Initially, the patient complained of progressive headaches, confusion, and weakness of the left arm. Magnetic resonance imaging of the brain showed a right temporoparietal tumor with substantial surrounding subcortical edema and midline shift to the left. Two consecutive craniotomies resulted in complete microsurgical resection of the lesion. Histology was consistent with a World Health Organization grade IV, IDH-wildtype glioblastoma. Further treatment was standard chemoradiation including intensity-modulated radiotherapy with oral temozolomide chemotherapy. Seven months after diagnosis, the cranial lesion progressed, and the patient developed painful metastases in multiple bones and suspicious right-sided cervical lymph nodes. Immunohistochemistry and molecular signature supported the case of a metastatic glioblastoma. Further treatment was palliative radiotherapy of the spinal lesions along with symptomatic pain management. Extracranial metastasis of glioblastoma is a rare complication of which only a few cases have been reported in the literature. Little is known about the precise mechanisms of tumor dissemination and the appropriate treatment., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany.)

Objective: To evaluate clinical results and long-term patient-reported outcome measures (PROMs) on quality of life in cervical cancer patients following radiochemotherapy (RCT) and brachytherapy (BT) as definitive treatment., Materials and Methods: Between 2003 and 2023, a total of 132 patients with advanced cervical cancer were evaluated for possible treatment. Patients treated by postoperative RCT, palliative radiotherapy, and those treated for recurrent disease were excluded. Thus, 46 patients receiving standard RCT and BT as their curative treatment were included in this study. PROMs were assessed prospectively by patients' self-completion of the EORTC-QLQ-C30 and EORTC-QLQ-CX24 questionnaires., Results: Five-year overall survival (OS), distant metastases-free survival (DMFS), and pelvic tumor-free survival rates (PTFS) were 53%, 54%, and 83%, respectively. A significant impact on OS was seen for FIGO (International Federation of Gynecologic Oncology) stage (IIB-IIIA: 79% vs. IIIB-IVA: 33%, p = 0.015), for overall treatment time (OTT; 50-65 d: 64% vs. > 65 d: 38%, p = 0.004), and for rectal D 2cc (≤ 73 Gy: 50% vs. > 73 Gy: 38%, p = 0.046). The identical parameters were significantly associated with DMFS (FIGO stage: p = 0.012, OTT: p = 0.008, D 2cc : p = 0.024). No parameters with a significant influence on PTFS were seen. In multivariate analysis, an impact of FIGO stage on OS (p = 0.05) and DMFS (p = 0.014) was detected, and of rectal D 2cc on DMFS (p = 0.031). The overall QoL score was 63/100. Cognitive function was the least impaired (84/100), while role functioning was the worst (67/100). On the symptom scale, insomnia (46/100), fatigue (41/100), dyspnea (32/100), pain (26/100), and financial difficulties (25/100) were scored the worst. According to EORTC-QLQ-CX24, peripheral neuropathy (36/100) and lymphedema (32/100) occurred most frequently. Impaired sexual/vaginal functioning (32/100) and body image (22/100) were also frequently recorded., Conclusion: In patients with advanced cervical cancer, a combination of RCT and BT remains an excellent treatment option. In terms of patient-reported long-term quality of life, specific support is needed to alleviate symptoms including lymphedema, peripheral neuropathy, and impaired sexual activity., (© 2024. The Author(s).)

This study aimed to evaluate the benefit of additional administration of oxaliplatin during fluorouracil-based neoadjuvant radiochemotherapy (nRCT) in terms of pathologic complete remission (pCR), disease-free survival (DFS), and overall survival (OS) in patients with advanced rectal cancer. Between 2006 and 2021, 669 patients (pts) were diagnosed with locally advanced rectal cancer, of whom a total of 414 pts with nRCT were identified and included in the study. A total of 283 pts were treated by nRCT using concurrent chemotherapy with fluorouracil or capecitabine; 131 pts were treated using a combination of fluorouracil or capecitabine and oxaliplatin. Propensity score matching analyses (PSM) with 114 pts in each group were used to balance the patients' characteristics. OS, DFS, pCR-rate, and potential prognostic factors were compared between the two groups. The median follow-up time was 59.5 weeks in the fluorouracil-group and 43 weeks in the fluorouracil/oxaliplatin group ( p = 0.003). After PSM, the pCR-rate (including sustained clinical complete remission) was 27% (31/114 pts) in the fluorouracil/oxaliplatin group and 16% (18/114 pts) in the fluorouracil-group ( p = 0.033). There was no difference between these two groups for both 10-year OS and DFS neither before nor after PSM, respectively (OS: 72.6% vs. 55.4%, p = 0.066, and 67.8% vs. 55.1%, p = 0.703, and DFS: 44.8% vs. 46.8%, p = 0.134, and 44.7% vs. 42.3%, p = 0.184). Multivariate analysis identified regression grading according to Dworak grade 4 (HR: 0.659; CI: 0.471-0.921; p = 0.015) and age over 60 years (HR: 2.231; CI: 1.245-4.001; p = 0.007) as independent predictors for OS. In conclusion, the addition of oxaliplatin to fluorouracil during nRCT significantly improved pCR-rate without having an impact on survival.

Background: Extracorporeal life support (ECLS) is a salvage treatment for acute circulatory failure. Our high-volume tertiary centre performs more than 100 implants annually and provides ECLS-transports. With this study, we aimed to analyze the incidence and risk factors of limb ischemia depending on the vascular access., Methods: Between January 1, 2007, and December 31, 2018, 937 patients received an ECLS. Preoperative, intraoperative, in-hospital and up to 5 years follow-up data were collected. Outcome measures were limb ischemia and survival., Results: In total, 402 femoro-femoral veno-arterial ECLS patients were identified. Mean age was 56 ± 16.7 years, 26.9% were female, 7.9% had a history of peripheral vascular disease. Cannulation was performed percutaneously in 82.1% (n = 330), surgically in 5.7% (n = 23) and combined in 12.2% (n = 49). Mortality was not significantly different between the groups (51.1% percutaneous, 43.5% surgical, 44.9% combined [p = 0.89]). There was no significant difference in limb ischemia either, but a trend toward an increased frequency in the percutaneous group (p = 0.0501). No amputation was necessary. Limb ischemia slightly increased in-hospital mortality (54.6%) but did not affect long-term survival beyond 30 days. Univariate analysis adjusted for cannulation methods revealed younger age and female gender as risk factors of limb ischemia and younger age for limb ischemia after percutaneous cannulation., Conclusions: Our study shows that percutaneous, surgical, and combined vascular access techniques for ECLS implantation are associated with comparable and low incidence of limb ischemia which slightly increases in-hospital mortality. Special precaution has to be taken in young and female patients., (© 2022 International Center for Artificial Organ and Transplantation (ICAOT) and Wiley Periodicals LLC.)

Background and Purpose: PD-1 and PD-L1 are involved in anticancer immunosurveillance, and their expression may be predictive for therapeutic effectiveness of specific antibodies. Their influence on response to neoadjuvant radiochemotherapy (RCT) and prognosis in patients with oesophageal adenocarcinoma (OAC) remains to be defined., Materials and Methods: Between 10/2004 and 06/2018, complete pre-RCT biopsy-specimens were available from 76 patients with locally advanced, non-metastatic OAC scheduled for trimodality therapy. We evaluated intra- and peritumoural expression of CD8, PD-1 and PD-L1 in pre-treatment specimens to determine their influence on tumour regression grade and survival. PD-1 and PD-L1 expression were considered positive (+) if ≥1% of all cells were stained positive, otherwise negative (-); densities of CD8+ cells were categorized as being high (Hi) or low (Lo) according to the median., Results: A negative PD-L1 expression in peritumoural cells predicted a poor tumour regression (RD 0.24 [95% CI 0.03-0.44], p = 0.023). A positive PD-1 expression in intra- as well as peritumoural cells was identified as an unfavourable prognostic factor (HR 0.52 [95% CI 0.29-0.93], p = 0.028; HR 0.50 [0.25-0.99], p = 0.047, respectively). With respect to CD8+ infiltration, positive PD-1 and PD-L1 expressions attenuated its favourable prognostic effect in intratumoural area (LoCD8/PD1 + vs. HiCD8/PD1-: HR 0.25 [0.09-0.69], p = 0.007; LoCD8/PDL1+ vs. HiCD8/PDL1-: HR 0.32 [0.12-0.89], p = 0.028) and were associated with negative outcome when seen in peritumoural area (HiCD8/PD1+ vs. LoCD8/PD1-: HR 0.29 [0.11-0.74], p = 0.010); HiCD8/PDL1+ vs. LoCD8/PDL1-: HR 0.33 [0.12-0.90], p = 0.031)., Conclusions: PD-1 and PD-L1 expression were identified to be of predictive and prognostic value in patients with OAC, particularly when considering CD8+ infiltration. Further validation by a large size dataset is required., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

Osteoarthritis is characterized by the loss of the articular cartilage, bone remodeling, pain, and disability. No pharmacological intervention can currently halt progression of osteoarthritis. Here, we show that blocking receptor tyrosine kinase-like orphan receptor 2 (ROR2) improves cartilage integrity and pain in osteoarthritis models by inhibiting yes-associated protein (YAP) signaling. ROR2 was up-regulated in the cartilage in response to inflammatory cytokines and mechanical stress. The main ligand for ROR2, WNT5A, and the targets YAP and connective tissue growth factor were up-regulated in osteoarthritis in humans. In vitro, ROR2 overexpression inhibited chondrocytic differentiation. Conversely, ROR2 blockade triggered chondrogenic differentiation of C3H10T 1 / 2 cells and suppressed the expression of the cartilage-degrading enzymes a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5. The chondrogenic effect of ROR2 blockade in the cartilage was independent of WNT signaling and was mediated by down-regulation of YAP signaling. ROR2 signaling induced G protein and Rho-dependent nuclear accumulation of YAP, and YAP inhibition was required but not sufficient for ROR2 blockade-induced chondrogenesis. ROR2 silencing protected mice from instability-induced osteoarthritis with improved structural outcomes, sustained pain relief, and without apparent side effects or organ toxicity. Last, ROR2 silencing in human articular chondrocytes transplanted in nude mice led to the formation of cartilage organoids with more and better differentiated extracellular matrix, suggesting that the anabolic effect of ROR2 blockade is conserved in humans. Thus, ROR2 blockade is efficacious and well tolerated in preclinical animal models of osteoarthritis., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Background and Purpose: Tumour infiltrating lymphocytes (TIL) and tumour associated macrophages (TAM) play a key role in anticancer immunosurveillance. We studied their influence on response to neoadjuvant radiochemotherapy (RCT) and prognosis in patients with oesophageal adenocarcinoma (OAC)., Materials and Methods: Between 10/2004 and 06/2018, pre-RCT biopsy-specimens were available from 76 patients with locally advanced, non-metastatic OAC scheduled for trimodality therapy. We evaluated intra- and peritumoural expression of FoxP3+-, CD8+-TIL and CD68+-, CD163+-TAM, contemplating cell density, cell ratios and cell-to-cell distances to determine a possible influence on tumour regression grade (TRG) and survival. Median follow-up time for all patients was 18 months (IQR 9-43), and 54 months (25-97) for surviving patients. Data were analysed using risk analysis, logrank test and Cox regression., Results: Poor tumour regression was detected for cN+ (RR 0.77 [95% CI 0.66-0.90], p = 0.001), low intratumoural FoxP3+/CD8+ ratio (RR 0.75 [0.60-0.96], p = 0.020), high peritumoural CD163+/CD68+ ratio (RR 0.77 [0.60-0.99], p = 0.045) and high intratumoural TAM density (RD -0.44 [-0.82 to -0.06], p = 0.023). Apart from poor resection quality and TRG, pretherapeutic high peritumoural CD8+ infiltration (HR 2.36 [1.21-4.61], p = 0.012) and short intratumoural FoxP3+ to CD8+ cell-to-cell distances in middle ranged CD8+ density (HR 2.55 [1.00-6.52], p = 0.050) were significant unfavourable prognostic factors in multivariate analysis., Conclusions: Immunologic parameters, such as CD8+-, FoxP3+-TIL and CD68+-, CD163+-TAM, were identified to be of independent predictive and prognostic value in patients with OAC. Further and independent validation of these biomarkers by a large size dataset may urgently be contemplated., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The current gold standard in peripheral nerve repair is nerve autografts for bridging gaps larger than a centimeter. However, autografts are associated with a low availability and the loss of function at the donor site. Nerve guidance conduits (NGCs) made of biocompatible and biodegradable materials reflect suitable alternatives. Clinically approved NGCs comprise either wraps that are rolled around the loose ends of the nerve or steady-state tubes; however, both lack internal guidance structures. Here, we established self-rolling NGCs to allow for gentle encapsulation of nerve cells together with supportive microenvironments, such as (1) an inner tube wall coating with a bioactive spider silk film, (2) an inner tube wall lining using an anisotropic spider silk non-woven mat, or (3) a luminal filler using an anisotropic collagen cryogel. Neuronal cells adhered and differentiated inside the modified tubes and formed neurites, which were oriented along the guidance structures provided by the spider silk non-woven mat or by the fibrillary structure of the collagen cryogel. Thus, our size-adaptable NGCs provide several features useful for peripheral nerve repair, and distinct combinations of the used elements might support and enhance the clinical outcome., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2020 The Authors.)

Growth and Differentiation Factor 5 (GDF5) is a key risk locus for osteoarthritis (OA). However, little is known regarding regulation of Gdf5 expression following joint tissue damage. Here, we employed Gdf5-LacZ reporter mouse lines to assess the spatiotemporal activity of Gdf5 regulatory sequences in experimental OA following destabilisation of the medial meniscus (DMM) and after acute cartilage injury and repair. Gdf5 expression was upregulated in articular cartilage post-DMM, and was increased in human OA cartilage as determined by immunohistochemistry and microarray analysis. Gdf5 expression was also upregulated during cartilage repair in mice and was switched on in injured synovium in prospective areas of cartilage formation, where it inversely correlated with expression of the transcriptional co-factor Yes-associated protein (Yap). Indeed, overexpression of Yap suppressed Gdf5 expression in chondroprogenitors in vitro. Gdf5 expression in both mouse injury models required regulatory sequence downstream of Gdf5 coding exons. Our findings suggest that Gdf5 upregulation in articular cartilage and synovium is a generic response to knee injury that is dependent on downstream regulatory sequence and in progenitors is associated with chondrogenic specification. We propose a role for Gdf5 in tissue remodelling and repair after injury, which may partly underpin its association with OA risk.

Encapsulation of enzymes is often necessary to stabilize them against environmental conditions or to protect them from other harmful enzymes such as proteases. Here, a refillable spatial confinement system was produced using a fully degradable self-rolling biopolymer bilayer. The enzyme containers comprise spider silk and chitosan and enable one-pot reactions in the micro- to milliliter regime by trapping the enzyme inside the semipermeable tube and allow the substrate and/or product either to diffuse freely or to be entrapped. The tubes are stable toward several organic and aqueous solvents. A one-tube system with esterase-2 was used to establish the system. Further, a two-tube system was applied to mimic enzymatic cascades, where the enzymes have to be separated, because they, for example, inhibit each other. The entrapment mode was also tested in the two-tube system, which is beneficial for toxic products or for obtaining high concentrations of the desired product.

Purpose: To retrospectively evaluate long-term treatment results following neoadjuvant chemoradiation (CRT) and radical surgery in patients with advanced adenocarcinoma (AC) of the oesophagus., Patients and Methods: Between 2005 and 2015, a total of 102 consecutive patients with a median age of 64 years (range, 44-86 years) and AC of the oesophagus were evaluated of whom 84 received a full CRT. A group of 51 patients was treated with neoadjuvant intent followed by radical surgery. A total dose of 50.4 Gy with mostly weekly paclitaxel/fluorouracil chemotherapy was administered. Six to eight weeks following CRT, a transthoracic subtotal oesophageal and proximal gastric resection was performed. Survival curves for overall survival and no evidence of disease (NED) survival (primary endpoints) were calculated according to Kaplan-Meier, and possible prognostic factors were evaluated by the log-rank test as well as by a Cox regression analysis., Results: Median follow-up time of the surviving patients was 48 months (range, 14-134 months). Overall and NED survival rates for patients of the study group (n = 51) were 40 and 32%, respectively, at 5 years. Age (p = 0.04), ypT category (p = 0.1) and the development of distant metastases (p = 0.05) were identified as (marginally) independent prognostic variables with impact on survival. Median survival time for patients of the study group (n = 51) was 45 ± 18 months (95%CI 9-81 months). Clear resection margins were achieved in 46/51 patients (92%). Regression rates with complete regression rare residual cancer and increased number of residual cells, but predominantly fibrosis were 33, 41, and 10%, respectively. Patterns of failure revealed local with distant recurrence in 2/51 (4%), regional recurrence alone in 2/51 (4%), and distant metastases in 27/51 (53%) patients., Conclusion: Neoadjuvant CRT in patients with AC of the oesophagus followed by thoracoabdominal surgery is a locally very effective concept. A significant tumour regression in almost 75% of the patients may stimulate prospective trials on the omission of radical surgery for some elderly patients. Due to a high rate of distant metastases further investigations in terms of effective systemic therapy may be warranted.

Objective and sensitive assessment of cartilage repair outcomes lacks suitable methods. This study investigated the feasibility of 3D ultrasound biomicroscopy (UBM) to quantify cartilage repair outcomes volumetrically and their correlation with established classification systems. 32 sheep underwent bilateral treatment of a focal cartilage defect. One or two years post-operatively the repair outcomes were assessed and scored macroscopically (Outerbridge, ICRS-CRA), by magnetic resonance imaging (MRI, MOCART), and histopathology (O'Driscoll, ICRS-I and ICRS-II). The UBM data were acquired after MRI and used to reconstruct the shape of the initial cartilage layer, enabling the estimation of the initial cartilage thickness and defect volume as well as volumetric parameters for defect filling, repair tissue, bone loss and bone overgrowth. The quantification of the repair outcomes revealed high variations in the initial thickness of the cartilage layer, indicating the need for cartilage thickness estimation before creating a defect. Furthermore, highly significant correlations were found for the defect filling estimated from UBM to the established classification systems. 3D visualisation of the repair regions showed highly variable morphology within single samples. This raises the question as to whether macroscopic, MRI and histopathological scoring provide sufficient reliability. The biases of the individual methods will be discussed within this context. UBM was shown to be a feasible tool to evaluate cartilage repair outcomes, whereby the most important objective parameter is the defect filling. Translation of UBM into arthroscopic or transcutaneous ultrasound examinations would allow non-destructive and objective follow-up of individual patients and better comparison between the results of clinical trials.

Tenosynovial giant cell tumour (TSGCT; synonym, pigmented villonodular synovitis (PVNS)) is a rare low-grade mesenchymal neoplasm of either intra-articular or extra-articular origin. The etiopathogenesis of TSGCT is still uncertain, but recent studies showed a translocation involving colony-stimulating factor 1 (CSF-1) gene in a subset of cases. Histological features mimicking TSGCT can sometimes be encountered in periprosthetic interface membranes. To investigate the frequency and morphologic spectrum of this phenomenon, we conducted a systematic analysis of 477 periprosthetic interface membranes and performed immunohistochemical analysis on a subset of lesions compared to genuine TSGCT. In 26 of 477 periprosthetic membrane samples (5 %), at least some TSGCT-like features were found and 18 cases (4 %) strongly resembled it. Wear particles were detected in 100 % of the TSGCT-like lesions but only in 63.3 % of the whole cohort of periprosthetic membranes (p value <0.001). Immunohistochemistry comparing true TSGCT and TSGCT-like membranes showed similar inflammatory infiltrates with slightly elevated CD3+/CD8+ T lymphocytes and a slightly higher proliferation index in TSGCT samples. In conclusion, TSGCT-like changes in periprosthetic membranes likely represent exuberant fibrohistiocytic inflammatory response induced by wear particles and should be distinguished from genuine (neoplastic) TSGCT. Although TSGCT and TSGCT-like periprosthetic membranes represent different entities, their comparable morphology might reflect analogous morphogenesis.

Introduction: Using hemoglobin concentration ([Hb]) to diagnose borderline iron deficiency and monitor the progress of its treatment is difficult because of the confounding effects of plasma volume. Because hemoglobin mass (Hbmass) is not affected by plasma volume, it may be a more sensitive parameter. The aim of this study was to monitor Hbmass, iron storage, and maximal oxygen consumption (V˙O2max) during and after oral iron therapy in subjects with severe and moderate iron deficiency., Methods: Three groups of female recreational athletes were monitored for at least 22 wk, as follows: 1) severe iron deficiency group (SID) (n = 8; ferritin, ≤12 ng·mL), 2) moderate iron deficiency group (MID) (n = 14; ferritin, ≤25 ng·mL), and 3) control group (n = 8; ferritin, >25 ng·mL). Hbmass and iron status were determined before, during, and up to 12 wk after at least 10 wk of oral iron supplementation. In total, five V˙O2max tests were performed before, during, and after the supplementation period., Results: Hbmass increased markedly in the SID group (15.6% ± 11.0%, P < 0.001) and slightly in the MID group (2.2% ± 3.7%, P < 0.05) by the end of the supplementation period and remained at this level for the following 12 wk. [Hb] and Hbmass were similarly affected, but Hbmass was more closely related to mean corpuscular volume and mean corpuscular hemoglobin than [Hb]. The SID group incorporated 534 ± 127 mg of iron into ferritin and hemoglobin, whereas the MID group incorporated 282 ± 68 mg of iron. V˙O2max increased only in the SID group by 0.20 ± 0.18 L·min (P < 0.05) and was closely related to Hbmass (P < 0.01)., Conclusions: Hbmass is a sensitive tool for monitoring recovery from iron deficiency anemia and assessing the effectiveness of iron supplementation in individuals with severe or moderate iron deficiency.

Hot thyroid nodules (HTNs) in children are rare. Their reported malignancy rate is higher than in adults. However molecular data are rare. We present clinical and molecular data for 33 consecutive (29 benign and 4 malignant) HTNs. 17/29 Benign HTNs (59%) harbored somatic TSHR mutations. The most commonly observed mutation was M453T (in 8/29 samples). T632I and D633Y mutations were each detected twice. All other TSHR mutations were each found in one sample, including the new A538T mutation. One NRAS mutation was detected in a benign HTN with a M453T mutation. A PAX8/PPARG rearrangement was found in one malignant HTN. A T632I mutation was detected in one hot papillary thyroid carcinoma. The percentage of TSHR mutation positive HTNs in children and adolescents is within the range observed in adults. Contrary to adults, the M453T mutation is the predominant TSHR mutation in HTNs of children and adolescents. The increased malignancy rate of HTNs of children does not appear to be associated with RAS, BRAF, PAX8/PPARG and RET/PTC mutations., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Objective: The pathophysiology of the most common joint disease, osteoarthritis (OA), remains poorly understood. Since synovial fluid (SF) bathes joint cartilage and synovium, we reasoned that a comparative analysis of its protein constituents in health and OA could identify pathways involved in joint damage. We undertook this study to perform a proteomic analysis of knee SF from OA patients and control subjects and to compare the results to microarray expression data from cartilage and synovium., Methods: Age-matched knee SF samples from 10 control subjects, 10 patients with early-stage OA, and 10 patients with late-stage OA were compared using 2-dimensional difference-in-gel electrophoresis and mass spectrometry (MS). MS with a multiplexed peptide selected reaction monitoring assay was used to confirm differential expression of a subset of proteins in an independent OA patient cohort. Proteomic results were analyzed by Ingenuity Pathways Analysis and compared to published synovial tissue and cartilage messenger RNA profiles., Results: Sixty-six proteins were differentially present in healthy and OA SF. Three major pathways were identified among these proteins: the acute-phase response signaling pathway, the complement pathway, and the coagulation pathway. Differential expression of 5 proteins was confirmed by selected reaction monitoring assay. A focused analysis of transcripts corresponding to the differentially present proteins indicated that both synovial and cartilage tissues may contribute to the OA SF proteome., Conclusion: Proteins involved in the acute-phase response signaling pathway, the complement pathway, and the coagulation pathway are differentially regulated in SF from OA patients, suggesting that they contribute to joint damage. Validation of these pathways and their utility as biomarkers or therapeutic targets in OA is warranted., (Copyright © 2013 by the American College of Rheumatology.)

Background: The use of predifferentiated mesenchymal stem cells (MSC) leads to better histological results compared with undifferentiated MSC in sheep. This raises the need for a longer term follow-up study and comparison with a clinically established method., Hypothesis: We hypothesized that chondrogenic in vitro predifferentiation of autologous MSC embedded in a collagen I hydrogel leads to better structural repair of a chronic osteochondral defect in an ovine stifle joint after 1 year. We further hypothesized that resulting histological results would be comparable with those of chondrocyte-seeded matrix-associated autologous chondrocyte transplantation (MACT)., Study Design: Controlled laboratory study., Methods: Predifferentiation period of ovine MSC within collagen gel in vitro was defined by assessment of several cellular and molecular biological parameters. For the animal study, 2 osteochondral lesions (7-mm diameter) were created at the medial femoral condyles of the hind legs in 9 sheep. Implantation of MSC gels was performed 6 weeks after defect creation. Thirty-six defects were divided into 4 treatment groups: (1) chondrogenically predifferentiated MSC gels (pre-MSC gels), (2) undifferentiated MSC gels (un-MSC gels), (3) MACT gels, and (4) untreated controls (UC). Histological, immunohistochemical, and radiological evaluations followed after 12 months., Results: After 12 months in vivo, pre-MSC gels showed significantly better histological outcome compared with un-MSC gels and UC. Compared with MACT gels, the overall scores were higher for O'Driscoll and International Cartilage Repair Society (ICRS). The repair tissue of the pre-MSC group showed immunohistochemical detection of interzonal collagen type II staining. Radiological evaluation supported superior bonding of pre-MSC gels to perilesional native cartilage. Compared with previous work by our group, no degradation of the repair tissue between 6 and 12 months in vivo, particularly in pre-MSC gels, was observed., Conclusion: Repair of chronic osteochondral defects with collagen hydrogels composed of chondrogenically predifferentiated MSC shows no signs of degradation after 1 year in vivo. In addition, pre-MSC gels lead to partially superior histological results compared with articular chondrocytes., Clinical Relevance: The results suggest an encouraging method for future treatment of focal osteochondral defects without donor site morbidity by harvesting articular chondrocytes.

Degenerative disorders of the musculoskeletal system, in particular osteoarthritis, are among the most common diseases of the elderly and their importance in an aging society is continuously increasing. Correspondingly, many surgical interventions are undertaken and pathological specimens submitted for histopathologic workup. The pathophysiology of osteoarthritis, which ultimately leads to joint destruction, is still poorly understood. The question remains as to whether the cause lies (mainly) within the chondrocytes themselves (e.g. cellular aging/senescence) or whether the synovial membrane or the subchondral bone are equally or even more important factors. The process of joint destruction can be evaluated in terms of pathogenesis (typing), extent (staging) and degree of the most extensive focal damage (grading). Because of the heterogeneity of the disease and substantial individual differences in progression, classification and grading of cartilage degeneration represents a complex task. Any pathology report should be concise and delineate only the essential features. Differentiating between primary osteoarthritis and secondary degenerative changes, e.g., due to previously unknown rheumatoid disease, bone necrosis or an infection of the joint, is of particular clinical interest.