Sortilin Is Upregulated in Osteoarthritis-Dependent Cartilage Calcification and Associated with Cellular Senescence.

Osteoarthritis (OA) is a chronic joint disease characterized by articular cartilage calcification, loss of articular cartilage, bone changes, pain, and disability. Cartilage calcification is one hallmark of OA and is predominantly caused by basic calcium crystals formed due to an imbalance of the pyrophosphate pathway. Sortilin is a transmembrane protein that contributes to vascular calcification in atherosclerosis by externalizing alkaline phosphatase (ALP)-containing vesicles. Calcification in atherosclerosis and osteoarthritis has been associated with cellular senescence. The aim of this study was to investigate the potential role of sortilin and senescence in osteoarthritis-dependent cartilage calcification. Osteoarthritic cartilage from human knee joints was collected after joint replacement, and samples were analyzed by immunohistochemistry and quantitative RT-PCR analysis. Human chondrocytes were treated with osteogenic medium for up to 21 days to induce calcification. Western blots for sortilin and ALP, as well as an ALP activity assay, were performed. Human chondrocytes were treated with mitomycin C to induce senescence, and sortilin expression was quantified at the protein and gene levels. Sections of knee joints from a murine model of osteoarthritis were stained for sortilin and p16 and analyzed by immunohistochemistry. Treatment of wild-type chondrocytes using an osteogenic medium similar to human chondrocytes was performed. Osteoarthritic cartilage from mouse and human knee joints showed an increased number of sortilin and p16-positive chondrocytes compared to healthy cartilage. This observation was corroborated by increased gene expression of sortilin and p16 in mild and moderate osteoarthritic cartilage samples. To investigate the mechanism of sortilin regulation, human chondrocytes were treated with osteogenic medium to induce calcification. Sortilin protein levels and expression were increased after 7 days of stimulation, whereas ALP levels and activity were upregulated after 21 days of stimulation. Similar observations were made in a murine osteoarthritis model. Mechanistically, senescent chondrocytes induced by mitomycin C showed an upregulation of sortilin and ALP gene expression compared to non-senescent chondrocytes. Our data indicate that sortilin and ALP are upregulated during cartilage calcification, which is associated with chondrocyte senescence and thus might contribute to the pathogenesis of osteoarthritis. Cellular senescence seems to induce sortilin expression.

Control of a COVID-19 outbreak in a nursing home by general screening and cohort isolation in Germany, March to May 2020.

Elderly care facilities have become a major focus of coronavirus disease (COVID-19) control. Here, we describe an outbreak of COVID-19 in a nursing home in Germany from 8 March to 4 May 2020 (58 days), and the effect of an intervention of general screening and cohort isolation. COVID-19 cases among residents and staff were recorded on a daily basis from the first positive SARS-CoV-2 test from a resident on 8 March 2020, until 4 May 2020 when the last staff member was classified COVID-19 negative. Eighty of 160 residents (50%) and 37 of 135 staff members (27%) tested positive for SARS-CoV-2. Twenty-seven of the 80 residents were asymptomatic but tested positive during the first general screening. Cohort isolation of SARS-CoV-2 positive residents by reorganising the facility proved to be a major effort. After the intervention, four further asymptomatic residents tested positive in follow-up screenings within a period of 6 days, and were possibly infected prior to the intervention. Thereafter, no further infections were recorded among residents. The described outbreak was controlled by implementing general screening and rigorous cohort isolation, providing a blueprint for similar facilities.

Autologous fibroblast transplantation at the vesico-ureteral junction as potential reconstructive cell replacement in an animal model.

PURPOSE: To evaluate the cellular survival of donor fibroblasts after transplantation at the vesico-ureteral junction (VUJ) and to analyse their potential for reconstructive cell replacement in an animal model as autologous fibroblasts have been used as soft tissue augmentation material for scared and damaged tissue. METHODS: Muscles biopsies were procured from the lower limb muscles of 4 pigs; cytoplasm of fibroblasts was labelled with nano-sized iron oxide particles. Six weeks after taking of the muscle biopsies, fibroblast transplantation was performed, 3 × 10(6) cells suspended in transplantation medium (in 1-ml syringes) were injected at the VUJ using the modified STING technique. Animals were killed 8 weeks later; seeded fibroblasts were identified using prussian blue staining protocol; histological evaluation and morphological analysis were performed by light microscopy (Mayer's haematoxylin-eosin staining); and bladders were scanned by MRI for visualization and localization of the iron-labelled donor cells. RESULTS: Donor fibroblast cell colonization and cellular viability at the VUJ was demonstrated by MRI and histochemically indicating cellular uptake of iron particles at the VUJ. It was also evident that transplanted fibroblasts integrate into the extracellular matrix of the distal ureter augmenting ureteral host tissue. CONCLUSIONS: Labelled implanted autologous fibroblasts were visualized by staining procedure as well as MRI scan demonstrating persistence at the VUJ, suggesting that in vitro expanded fibroblasts survived in vivo after transplantation.

Molecular characteristics and alterations during early development of the human vagina.

Unresolved questions remain concerning the derivation of the vagina with respect to the relative contributions from the Müllerian ducts, the urogenital sinus, and the Wolffian ducts. Recent molecular and cellular studies in rodents have opened up a large gap between the level of understanding of vaginal development in mice and understanding of human vaginal development, which is based on histology. To compare the findings in mice with human vaginal development and to address this gap, we analysed molecular characteristics of the urogenital sinus, Wolffian ducts, and Müllerian ducts in 8-14-week-old human specimens using immunohistochemical methods. The monoclonal antibodies used were directed against cytokeratin (CK) 14, CK19, vimentin, laminin, p63, E-cadherin, caspase-3, Ki67, HOX A13, and BMP-4. The immunohistochemical analysis revealed that, during weeks 8-9, the epithelium of the Müllerian ducts became positive for p63 as p63-positive cells that originated from the sinus epithelium reached the caudal tip of the fused Müllerian ducts via the Wolffian ducts. The lumen of the fused Müllerian ducts was closed by an epithelial plug that contained both vimentin-positive and vimentin-negative cells. Subsequently, the resulting epithelial tube enlarged by proliferation of basal p63-positive cells. The first signs of squamous differentiation were detected during week 14, with the appearance of CK14-positive cells. According to our results, all three components, namely, the urogenital sinus, Wolffian ducts, and Müllerian ducts, interacted during the formation of the human vagina. The sinus epithelium provided p63-positive cells, the Wollfian ducts acted as a 'transporter', and the Müllerian ducts contributed the guiding structure for the vaginal anlagen. Epithelial differentiation began at the end of the period studied and extended in a caudo-cranial direction. The present study is one of the first to provide up-to-date molecular correlates for human vaginal development that can be compared with the results of cell biological studies in rodents.

Phytotoxicity of wastewater-born micropollutants--Characterisation of three antimycotics and a cationic surfactant.

Sewage sludge applied to soil may be a valuable fertiliser but can also introduce poorly degradable and highly adsorptive wastewater-born residues of pharmaceuticals and personal care products (PPCPs) to the soil, posing a potential risk to the receiving environment. Three azole antimycotics (climbazole, ketoconazole and fluconazole), and one quaternary ammonium compound (benzyldimethyldodecylammonium chloride, BDDA) that are frequently detected in municipal sewage sludge and/or treated wastewater were therefore characterised in their toxicity toward terrestrial (Brassica napus) and aquatic (Lemna minor) plants. Fluconazole and climbazole showed the greatest toxicity to B. napus, while toxicity of ketoconazole and BDDA was by one to two orders of magnitude lower. Sludge amendment to soil at an agriculturally realistic rate of 5 t/ha significantly reduced the bioconcentration of BDDA in B. napus shoots compared to tests without sludge amendment, although not significantly reducing phytotoxicity. Ketoconazole, fluconazole and BDDA proved to be very toxic to L. minor with median effective concentrations ranging from 55.7 µg/L to 969 µg/L. In aquatic as well as terrestrial plants, the investigated azoles exhibited growth-retarding symptoms presumably related to an interference with phytohormone synthesis as known for structurally similar fungicides used in agriculture. While all four substances exhibited considerable phytotoxicity, the effective concentrations were at least one order of magnitude higher than concentrations measured in sewage sludge and effluent. Based on preliminary hazard quotients, BDDA and climbazole appeared to be of greater environmental concern than the two pharmaceuticals fluconazole and ketoconazole.

Results of extended plant tests using more realistic exposure scenarios for improving environmental risk assessment of veterinary pharmaceuticals.

BACKGROUND: Residues of veterinary medicinal products (VMPs) enter the environment via application of manure onto agricultural areas where in particular antibiotics can cause phytotoxicity. Terrestrial plant tests according to OECD guideline 208 are part of the environmental risk assessment of VMPs. However, this standard approach might not be appropriate for VMPs which form non-extractable residues or transformation products in manure and manure-amended soil. Therefore, a new test design with a more realistic exposure scenario via manure application is needed. This paper presents an extended plant test and its experimental verification with the veterinary antibiotics florfenicol and tylosin tartrate. With each substance, plant tests with four different types of application were conducted: standard tests according to OECD 208 and three tests with application of test substance via spiked manure either without storage, aerobically incubated, or anaerobically incubated for different time periods. RESULTS: In standard tests, the lowest NOEC was <0.06 mg/kg dry soil for florfenicol and 16.0 mg/kg dry soil for tylosin tartrate. Pre-tests showed that plant growth was not impaired at 22-g fresh manure/kg dry soil, which therefore was used for the final tests. The application of the test substances via freshly spiked as well as via aerobically incubated manure had no significant influence on the test results. Application of florfenicol via anaerobically incubated manure increased the EC10 by a factor up to 282 and 540 for half-maximum and for maximum incubation period, respectively. For tylosin tartrate, this factor amounted to 64 at half-maximum and 61 at maximum incubation period. The reduction of phytotoxicity was generally stronger when using cattle manure than pig manure and particularly in tests with cattle manure phytotoxicity decreased over the incubation period. CONCLUSIONS: The verification of the extended plant test showed that seedling emergence and growth are comparable to a standard OECD 208 test and reliable effect concentrations could be established. As demonstrated in the present study, phytotoxicity of veterinary antibiotics can be significantly reduced by application via incubated manure compared to the standard plant test. Overall, the presented test design proved suitable for inclusion into the plant test strategy for VMPs.

Assessing the ecological long-term impact of wastewater irrigation on soil and water based on bioassays and chemical analyses.

The reuse of treated wastewater for irrigation and groundwater recharge can counteract water scarcity and reduce pollution of surface waters, but assessing its environmental risk should likewise consider effects associated to the soil. The present study therefore aimed at determining the impact of wastewater irrigation on the habitat quality of water after soil passage and of soil after percolation by applying bioassays and chemical analysis. Lab-scale columns of four different soils encompassing standard European soil and three field soils of varying characteristics and pre-contamination were continuously percolated with treated wastewater to simulate long-term irrigation. Wastewater and its percolates were tested for immobilization of Daphnia magna and growth inhibition of green algae (Pseudokirchneriella subcapitata) and water lentils (Lemna minor). The observed phytotoxicity of the treated wastewater was mostly reduced by soil passage, but in some percolates also increased for green algae. Chemical analysis covering an extensive set of wastewater-born organic pollutants demonstrated that many of them were considerably reduced by soil passage, particularly through peaty soils. Taken together, these results indicated that wastewater-born phytotoxic substances may be removed by soil passage, while existing soil pollutants (e.g. metals) may leach and impair percolate quality. Soils with and without wastewater irrigation were tested for growth of plants (Avena sativa, Brassica napus) and soil bacteria (Arthrobacter globiformis) and reproduction of collembolans (Folsomia candida) and oligochaetes (Enchytraeus crypticus, Eisenia fetida). The habitat quality of the standard and two field soils appeared to be deteriorated by wastewater percolation for at least one organism (enchytraeids, plants or bacteria), while for two pre-contaminated field soils it also was improved (for plants and/or enchytraeids). Wastewater percolation did not seem to raise soil concentrations of classical organic pollutants and priority substances, while a significant retention was found for zinc and several organic micropollutants, particularly in the peaty soils, thus matching these soils' observed higher removal efficiency. Overall, our results demonstrate that benefits of wastewater irrigation can come with the cost of deteriorating soil habitat quality and depend on the respective soil and considered test organism. The approach employed here represents a feasible tool to assess these integrated effects at lab-scale while being predictive for scenarios at field-scale.

Ecotoxicity of climbazole, a fungicide contained in antidandruff shampoo.

Emerging pollutants such as personal care products can reach the environment via effluents from wastewater treatment plants (WWTPs) and digested sludge. Only recently, the antidandruff agent and antimycotic climbazole was detected for the first time in a WWTP effluent with concentrations up to 0.5 µg/L. Climbazole acts as a C14-demethylase inhibitor (DMI) fungicide and thus has a high efficacy against fungi, but knowledge of its potential environmental impact is lacking. Therefore, the aim of the present study was to characterize climbazole's ecotoxicity by conducting standard biotests with organisms representing different trophic levels from the aquatic as well as the terrestrial ecosystems. It was found that the toxicity of climbazole is mostly similar to that of other DMI fungicides, whereas it proved to be particularly toxic to primary producers. The lowest median effective concentrations (EC50s) were determined for Lemna minor, at 0.013 mg/L (biomass yield), and Avena sativa, at 18.5 mg/kg soil dry weight (shoot biomass). Reduction of frond size in water lentils and shoot length in higher plants suggested an additional plant growth-retarding mode of action of climbazole. In addition, it was demonstrated here that for an ionizable compound such as climbazole, the soil pH can have a considerable influence on phytotoxicity.

[Forensic-psychiatric opinion at the beginning of the 19th century. Theory and practice in Johann Christian August Heinroth (1773-1843)]. / Forensisch-psychiatrische Begutachtung zu Beginn des 19. Jahrhunderts: Theorie und Praxis bei Johann Christian August Heinroth (1773-1843).

OBJECTIVE: In recent years, Leipzig psychiatrist Johann Christian August Heinroth has attracted more and more attention, yet his forensic-psychiatric works have largely been neglected. METHODS: Therefore this study takes a close look at his System der Psychisch-Gerichtlichen Medizin (1825) and his Gerichtsärztliche- und Privatgutachten (1847) to examine his theoretical and practical approach to examining offenders. RESULTS AND CONCLUSIONS: Heinroth wanted to give forensic-psychiatric opinion on the responsibility of specialist physicians and to implement training for them. He also set up firm criteria for reports, both as regarded content and form. In such reports, he made a full review of the culprit's medical history, present physical and mental state and capabilities, as well as of any influences that might have had an impact on the deed. He also ruled that a specialist's report was to help the judge, but not to replace the judge's work. Heinroth's own works reveal his ideas, as well as his rejection of the exploitation of (assumed) mental disorders to gain exculpation for offenders.

The role of cartilage canals in endochondral and perichondral bone formation: are there similarities between these two processes?

We investigated the development of cartilage canals to clarify their function in the process of bone formation. Cartilage canals are tubes containing vessels that are found in the hyaline cartilage prior to the formation of a secondary ossification centre (SOC). Their exact role is still controversial and it is unclear whether they contribute to endochondral bone formation when an SOC appears. We examined the cartilage canals of the chicken femur in different developmental stages (E20, D2, 5, 7, 8, 10 and 13). To obtain a detailed picture of the cellular and molecular events within and around the canals the femur was investigated by means of three-dimensional reconstruction, light microscopy, electron microscopy, histochemistry and immunohistochemistry [vascular endothelial growth factor (VEGF), type I and II collagen]. An SOC was visible for the first time on the last embryonic day (E20). Cartilage canals were an extension of the vascularized perichondrium and its mesenchymal stem cell layers into the hyaline cartilage. The canals formed a complex network within the epiphysis and some of them penetrated into the SOC were they ended blind. The growth of the canals into the SOC was promoted by VEGF. As the development progressed the SOC increased in size and adjacent canals were incorporated into it. The canals contained chondroclasts, which opened the lacunae of hypertrophic chondrocytes, and this was followed by invasion of mesenchymal cells into the empty lacunae and formation of an osteoid layer. In older stages this layer mineralized and increased in thickness by addition of further cells. Outside the SOC cartilage canals are surrounded by osteoid, which is formed by the process of perichondral bone formation. We conclude that cartilage canals contribute to both perichondral and endochondral bone formation and that osteoblasts have the same origin in both processes.

Impacto do serviço de patologia na prevalência do câncer de colo uterino em saúde pública

Objetivo: Avaliar a prevalência do carcinoma invasor e ?in situ? de colo uterino pré e pós a implantação do serviço de patologia cervical na saúde pública de Joinville. Métodos: Estudo transversal descritivo retrospectivo.Foram estudados todos os resultados histopatológicos positivos para câncer de colo de útero invasor e ?in situ?, obtidos através da busca nos dois laboratórios de análise patológica que servem à saúde pública de Joinville, SC. O período do estudo contemplou 22 anos, e foi diferenciadoem dois tempos. O primeiro foi de janeiro de 1985 a dezembro de 1995, e o segundo de janeiro de 1996 a dezembro de 2006, sendo o marco do segundo período a implantação do ambulatório de patologia do colo deútero em janeiro de 1996 no serviço público de Joinville, SC. Os cálculos estatísticos foram realizados no programa SPSS. Para se testar a homogeneidade dos grupos em relação às proporções foi utilizado o teste qui-quadrado. Resultados: Foram estudados 788 laudos de histopatológicos positivos para carcinoma ?in situ? e invasor de colo uterino. Observou-se que a prevalência média de carcinoma ?in situ? entre 1985 e 1995 foi de 42 casos por 100.00 habitantes e entre 1996 e 2006, 85 por 100.000 habitantes. Com relação ao carcinoma invasor, a prevalência média foi 192 casos por 100.000 habitantes no primeiro período e no segundo período, 182 casos por 100.000 habitantes.Conclusões: Conclui-se que houve diminuição das formas avançadas e aumento das formas precursoras de câncer de colo uterino após a implantação de serviço de referência.

Objective: To evaluate the prevalence of ?in situ? and invasive carcinoma before and after the deployment of cervical pathology department in the public health in Joinville. Methods: This was a retrospective descriptive crosssectional study. All histopathological positive results for cervix carcinoma, invasive and ?in situ? were studied obtained by two laboratories in search of pathological analysis that serve the public health in Joinville, SC. The period of the study included 22 years, and was differentiated into two phases. The first was from January 1985 to December 1995, and the second from January 1996 to December 2006, being the mark of the second period the deployment of departament?s cervical pathology in January 1996 in the public service of Joinville, SC . The statistical calculations were performed in SPSS. To test the homogeneity of the groups for proportions was used chi-square. Results: We studied 788 reports of histopathological positive for cervix carcinoma in situ and invasive. In examining the two periods, it was observed that theaverage prevalence of carcinoma ?in situ? between 1985 and 1995 was 42 cases per 100.00 inhabitants and between 1996 and 2006, 85 per 100,000 inhabitants. With respect to invasive carcinoma, the average prevalence was 192 cases per 100,000 inhabitants in the first periodand second period, 182 cases per 100,000 inhabitants. Conclusions: We conclude that there was a decrease in advanced forms and ways of increasing in precursor cervical cancer after the deployment of service reference.

Leiomioma atípico de útero: relato de caso / Atypical uterus Leiomioma: case report

O leiomioma atípico, também conhecido como leiomioma bizarro ou simplástico, é uma neoplasia do músculo liso do útero que contém uma variedade de células musculares lisas gigantes com múltiplos núcleos pleomórficos e com cromatina grumosa. Seu potencial biológico é determinado pelo índice mitótico e necrose tumoral, estimando-se como malignos aqueles com áreasde necrose ou aqueles com 10 ou mais figuras de mitoses por 10 campos de grande aumento. Considerando um novo caso de leiomioma atípico de útero e revisão de literatura, os aspectos clínicos e patológicos dessa neoplasia são descritos...

Criterios de selección para el tratamiento del linfedema / Criteria in choosing treatment of lymphoedema

Objetivo: Fundamentar los criterios de selección del tratamiento médico y/o quirúrgico en base a 180 pacientes con linfedema. Material y métodos: La linfografía radioisotópica dinámica se realizó siempre en los linfedemas secundarios con menos de 2 años de evolución y cuando se sospechó megalinfáticos. El eco-doppler color venoso se realizó sistemáticamente. Se efectuaron: 1 operación de Thompson modificada acorde en linfidema primario; 19 anastomosis linfo-venosas en linfidema secundario; 8 resecciones e injertos libres de piel en linfidema genital; 1 operación de Servelle. Ciento cincuenta y un pacientes fueron seleccionados para terapia médica descongestiva compleja como único tratamiento. Resultados: No se repitieron las crisis linfangíticas. El linfadema fue de más fácil control con tratamiento médico en su evolución alejada. Controló en este período el linfedema, sin recidiva. Permitió la deambulación. Los volúmenes se redujeron significativamente, acorde con la cooperación del paciente. Conclusiones: El tratamiento médico es la base de la terapia en linfedema. La cirugía tiene indicación: 1) en linfedemas gigantes (exéresis) para permitir la deambulación y controlar las crisis linfangíticas; 2) las anastomosis linfo-venosas son patrimonio de los linfedemas secundarios con vías funcionantes; 3) en el linfedema genital masculino la cirugía es el mejor tratamiento (AU)

Criterios de selección para el tratamiento del linfedema / Criteria in choosing treatment of lymphoedema

Objetivo: Fundamentar los criterios de selección del tratamiento médico y/o quirúrgico en base a 180 pacientes con linfedema. Material y métodos: La linfografía radioisotópica dinámica se realizó siempre en los linfedemas secundarios con menos de 2 años de evolución y cuando se sospechó megalinfáticos. El eco-doppler color venoso se realizó sistemáticamente. Se efectuaron: 1 operación de Thompson modificada acorde en linfidema primario; 19 anastomosis linfo-venosas en linfidema secundario; 8 resecciones e injertos libres de piel en linfidema genital; 1 operación de Servelle. Ciento cincuenta y un pacientes fueron seleccionados para terapia médica descongestiva compleja como único tratamiento. Resultados: No se repitieron las crisis linfangíticas. El linfadema fue de más fácil control con tratamiento médico en su evolución alejada. Controló en este período el linfedema, sin recidiva. Permitió la deambulación. Los volúmenes se redujeron significativamente, acorde con la cooperación del paciente. Conclusiones: El tratamiento médico es la base de la terapia en linfedema. La cirugía tiene indicación: 1) en linfedemas gigantes (exéresis) para permitir la deambulación y controlar las crisis linfangíticas; 2) las anastomosis linfo-venosas son patrimonio de los linfedemas secundarios con vías funcionantes; 3) en el linfedema genital masculino la cirugía es el mejor tratamiento