A 13-year patient journey of infant giant clival chordoma: case report and literature review.

Chordomas are rare malignant bone tumours that develop from the ectopic remnants of the embryonic notochord. In contrast to adults, the majority in children under 16 present intra-cranially (63%). In 2006, we reported the youngest case of a large clival chordoma, a 15-week old baby, the second case to present without skull base involvement and the fourth case of chordoma in a patient with tuberous sclerosis (TS) Kombogiorgas (Childs Nerv Syst 22(10):1369-1374, 2006). In this report, we provide an update on this patient's journey through a range of therapeutic options and summarize an update of the literature, since 2006, for this patient group.

African swine fever: a New Zealand perspective on epidemiological risk factors for its occurrence.

This article reviews key epidemiological and clinical features of African swine fever (ASF). We identify particular aspects of New Zealand's pig populations (commercial, non-commercial, and wild) that may affect the risk of disease entry or spread. Review of published literature is supplemented by analysis of demographic and spatial aspects of the New Zealand commercial, non-commercial, and feral pig populations to provide context around risk factors for the disease that are most relevant to New Zealand. The current Eurasian outbreak of ASF, including recent spread into Oceania, has increased the risk of an incursion of the disease into New Zealand. Large volumes of fresh pork importation (including from countries affected by ASF), large non-commercial pig populations with substantial spatial overlap with the country's commercial industry, limited monitoring of compliance with waste food feeding regulations, and lack of mandatory premises identification for non-commercial pig holdings would likely contribute to the risk of spread of ASF in the event of an incursion. Awareness amongst veterinarians of these risk factors will contribute to national biosecurity and disease preparedness efforts in New Zealand.

Probiotic effect of Bifidobacterium longum 51A and Weissella paramesenteroides WpK4 on gerbils infected with Giardia lamblia.

AIMS: The objective of this study was to assess the probiotic potential of genuine strains of Bifidobacterium longum 51A and Weissella paramesenteroides WpK4, in experimental giardiasis. METHODS AND RESULTS: The bacteria were administered orally to gerbils (Meriones unguiculatus) 10 days before oral infection with trophozoites of Giardia lamblia. After 7 days of infection, the animals were euthanized and portions of the duodenum were processed for histopathologic, histochemical and morphometric assessment. The height of the intestinal crypts and crypt/villi ratio were higher in infected groups (P < 0·05) than in noninfected groups. The area of mucus production was higher (P < 0·05) in infected animals pretreated with B. longum 51A than in other groups. The parasitic load of the animals that received both bacteria decreased significantly (P < 0·05) compared to the ones of the control group. CONCLUSIONS: Our results suggest a probiotic function of B. longum 51A and W. paramesenteroides WpK4 and may result in their use as a prophylactic and therapeutic alternative for promoting human and animal health. SIGNIFICANCE AND IMPACT OF THE STUDY: Bifidobacterium longum 51A and W. paramesenteroides WpK4 may constitute prophylactic alternatives, reversing the emergence of side effects and resistance observed in the conventional treatment of giardiasis.

Visfatin alters the cytokine and matrix-degrading enzyme profile during osteogenic and adipogenic MSC differentiation.

OBJECTIVES: Age-related bone loss is associated with bone marrow adiposity. Adipokines (e.g., visfatin, resistin, leptin) are adipocyte-derived factors with immunomodulatory properties and might influence differentiation of bone marrow-derived mesenchymal stem cells (MSC) in osteoarthritis (OA) and osteoporosis (OP). Thus, the presence of adipokines and MMPs in bone marrow and their effects on MSC differentiation were analyzed. METHODS: MSC and ribonucleic acid (RNA) were isolated from femoral heads after hip replacement surgery of OA or osteoporotic femoral neck fracture (FF) patients. Bone structural parameters were evaluated by microcomputed tomography (µCT). MSC were differentiated towards adipocytes or osteoblasts with/without adipokines. Gene expression (adipokines, bone marker genes, MMPs, TIMPs) and cytokine production was evaluated by realtime-polymerase chain reaction (realtime-PCR) and enzyme-linked immunosorbent assay (ELISA). Matrix mineralization was quantified using Alizarin red S staining. RESULTS: µCT showed an osteoporotic phenotype of FF compared to OA bone (reduced trabecular thickness and increased ratio of bone surface vs volume of solid bone). Visfatin and leptin were increased in FF vs OA. Visfatin induced the secretion of IL-6, IL-8, and MCP-1 during osteogenic and adipogenic differentiation. In contrast to resistin and leptin, visfatin increased MMP2 and MMP13 during adipogenesis. In osteogenically differentiated cells, MMPs and TIMPs were reduced by visfatin. Visfatin significantly increased matrix mineralization during osteogenesis, whereas collagen type I expression was reduced. CONCLUSION: Visfatin-mediated increase of matrix mineralization and reduced collagen type I expression could contribute to bone fragility. Visfatin is involved in impaired bone remodeling at the adipose tissue/bone interface through induction of proinflammatory factors and dysregulated MMP/TIMP balance during MSC differentiation.

Ultrastructure aspects of Brycon gouldingi (Teleostei, Characidae) related to swimming ability and feeding during larval development.

The larval ultrastructure of Brycon gouldingi related to swimming and feeding from hatching to total yolk absorption is described from scanning electron micrographs. Newly hatched larvae (time zero) had no mouth opening, undefined optic vesicles, an olfactory plate visible as a shallow depression, rudimentary gill arches, neural groove, embryonic fin and a primary neuromast in the dorsal region of the head. At the time of yolk absorption, 55 h post hatching, the larvae presented an optic vesicle comprising an optic cup and crystalline lens; a mouth with tongue, tapered teeth and taste buds; a ciliated olfactory cavity; branched gill arches; filled neural groove signalling central nervous system development; caudal, pectoral, dorsal and anal fins; and neuromasts distributed throughout the head and body. These characters are related to prey capture and swimming ability, key aspects of survival during the larval stage. The results of this study provide important information for exploitation and aquaculture of B. gouldingi.

[Rheumatoid arthritis]. / Rheumatoide Arthritis.

Rheumatoid arthritis (RA) is a chronic and progressive systemic disease of the connective tissue, which is particularly manifested with destructive alterations to the joints. Inflammatory reactions in the synovium lead to the influx of peripheral inflammatory cells as well as the activation of local cells. Released growth factors, chemokines and especially cytokines play a key role in chronic inflammatory responses. In addition to the central lymphocytes, the T and B cells and their subpopulations, locally resident cells, such as neutrophils, macrophages and fibroblasts as well as cells of bone metabolism are activated by the inflammatory milieu and contribute to and drive inflammation and tissue damage. The destruction of cartilage and bone substance by local tissue cells, synovial fibroblasts and osteoclasts is characteristic for this disease. Untreated, the local inflammatory and destructive processes as well as systemic inflammatory factors lead to progressive and irreversible joint destruction. Cellular and immunological processes in RA are closely interwoven; therefore, besides the general inhibition of immunological processes, specific inhibition of central key molecules can reduce or completely stop the inflammatory destructive processes; however, a high heterogeneity can be observed among RA patients and disease progression. Therefore, an expansion of the therapeutic options is desirable as not all patients are able to equally benefit from the therapeutic treatment. It is important to characterize new molecular mechanisms, which could lead to the development of new therapeutic options. Some of the more recent insights are summarized in this overview.

Selection of a candidate probiotic strain of Pediococcus pentosaceus from the faecal microbiota of horses by in vitro testing and health claims in a mouse model of Salmonella infection.

AIMS: The aim of this study was to verify the suitable use of candidate 'probiotics' selected by in vitro tests and the importance of in vivo assays to nominate micro-organisms as probiotics and alternative prophylactic treatments for Salmonella Typhimurium infection. METHODS AND RESULTS: Thirty-three lactic acid bacteria (LAB) isolated from foal's faeces were assessed based on the main desirable functional in vitro criteria. Based on these results, Pediococcus pentosaceus strain 40 was chosen to evaluate its putative probiotic features in a mouse model of Salmonella infection. Daily intragastric doses of Ped. pentosaceus 40 for 10 days before and 10 days after Salmonella challenge (106 CFU of Salm. Typhimurium per mouse) led to a significant aggravation in mouse health by increasing weight loss, worsening clinical symptoms and anticipating the time and the number of deaths by Salmonella. Pediococcus pentosaceus modulated cell-mediated immune responses by up-regulation of the gene expression of the proinflammatory cytokines IFN-γ and TNF-α in the small intestine. CONCLUSION: The usual criteria were used for in vitro screening of a large number of LAB for desirable probiotic functional properties. However, the best candidate probiotic strain identified, Ped. pentosaceus #40, aggravated the experimental disease in mice. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings emphasize the need for prophylactic or therapeutic effectiveness to be demonstrated in in vivo models to make precise health claims.

Ultra-thin resin embedding method for scanning electron microscopy of individual cells on high and low aspect ratio 3D nanostructures.

The preparation of biological cells for either scanning or transmission electron microscopy requires a complex process of fixation, dehydration and drying. Critical point drying is commonly used for samples investigated with a scanning electron beam, whereas resin-infiltration is typically used for transmission electron microscopy. Critical point drying may cause cracks at the cellular surface and a sponge-like morphology of nondistinguishable intracellular compartments. Resin-infiltrated biological samples result in a solid block of resin, which can be further processed by mechanical sectioning, however that does not allow a top view examination of small cell-cell and cell-surface contacts. Here, we propose a method for removing resin excess on biological samples before effective polymerization. In this way the cells result to be embedded in an ultra-thin layer of epoxy resin. This novel method highlights in contrast to standard methods the imaging of individual cells not only on nanostructured planar surfaces but also on topologically challenging substrates with high aspect ratio three-dimensional features by scanning electron microscopy.

[Inflammation and bone : Osteoimmunological aspects]. / Entzündung und Knochen : Osteoimmunologische Aspekte.

Microscopic fractures (so-called microcracks) or traumatic macrofractures require bone, as the basic scaffold of the human body, to have a high regenerative capability. In order to be able to provide this regenerative capability, bone is in a constant process of remodeling. This finely tuned homeostasis of bone formation and degradation can become disrupted, which leads to osteoporosis or other bone disorders. It has been shown that the immune system is substantially involved in the regulation of bone homeostasis and that chronic inflammation in particular can disturb this balance; therefore, this article reviews the osteoimmunological aspects contributing to osteoporosis and other diseases associated with bone degradation.

Morphological and morphometric aspects of early life stages of piabanha Brycon gouldingi (Characidae).

Adult specimens of piabanha Brycon gouldingi were collected from Rio das Mortes (Mato Grosso, Brazil), adapted to captivity and induced to spawn at Buriti Fisheries (Nova Mutum, MT, Brazil). The early developmental stages of B. gouldingi were then characterized. Samples were collected at pre-determined times from oocyte extrusion to total yolk absorption. Oocyte diameter, total larval length (LT ) and yolk-sac volume were measured. The mean ± s.d. duration of embryo developmental of B. gouldingi was 13·90 ± 0·06 h at 26·40 ± 1·13° C. The mean ± s.d. oocyte diameter was 1·13 ± 0·06 mm with 54% of oocytes ranging from 1·11 to 1·20 mm. Seven stages characterized the early developmental phase of this species: zygote, cleavage, morula, blastula, gastrula, histogenesis-organogenesis and hatching, with unique features related to each stage. At hatching, the larvae measured 3·40 ± 0·07 mm, presented an elongated shape with yolk-sac volume of 0·46 ± 0·08 µl, non-pigmented eyes and exhibited swimming ability. When the yolk was completely absorbed at 55 h post-hatch, mean ± larval LT was 6·68 ± 0·65 mm, the eyes were highly pigmented and the teeth were visible. These are the first reported findings on the initial developmental stages of B. gouldingi and could be used to improve captive breeding management and conservation practices.

[Fibroblasts as pathogenic cells in rheumatic inflammation]. / Fibroblasten als pathogene Zellen in der rheumatischen Entzündung.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by synovitis, synovial hyperplasia and progressive degeneration of affected joints. These processes are mediated by cells of the immune system as well as by synovial fibroblasts (RASF) originating from the lining layer of the synovium. In this scenario RASFs display an activated phenotype: they show an altered expression of adhesion molecules which allows attachment to articular cartilage and by synthesis of proteases they mediate progressive cartilage and bone destruction. Furthermore, they produce various cytokines and chemokines, which are essential for promoting the inflammatory response. In recent years it has become evident that RASFs not only passively respond to the proinflammatory milieu in the joints of RA patients but also actively contribute by the overproduction of several cytokines and chemokines. These proinflammatory cytokines trigger the transformation of RASFs into an aggressive and invasive phenotype. Additionally, the primarily altered genuine RASFs are actively involved in the recruitment and activation of immune cells. Taken together, they are key players in the development of the well-known chronic, destructive inflammatory response in joints affected by RA.

A pilot study on the effectiveness of a rose hip shell powder in patients suffering from chronic musculoskeletal pain.

We carried out a 3-month preliminary investigation on the effectiveness of a rose hip shell powder and its mechanism of action. Of 52 patients suffering from acute exacerbations of low back pain (n = 39) or knee pain (n = 13), 29 had participated earlier in the pilot study with the pseudofruit powder Litozin(®) . After assessing the baseline values, patients were offered up to 20 g of a rose hip shell powder per day. Patients were encouraged to adjust the daily dose upwards or downwards according to their symptoms for the period of 3 months. The examination for possible effectiveness was by intention-to-treat analysis with last observation carried forward. There was no difference in any generic or disease-specific outcome variables between the patients consuming the rose hip shell powder and those consuming the pseudofruit powder Litozin(®) in the previous surveillance study. A human protein array system and fractions from the rose powders were used to study their effect on cytokine expression in vitro. The data indicate that lipophilic rose hip fractions from the shell and the pseudofruit inhibit cytokine expression and that the shell powder may be the better starting material for a future rose hip extract prepared with a lipophilic solvent.

[Inflammation and bone metabolism]. / Entzündung und Knochenmetabolismus.

A finely balanced relationship between bone resorption and bone formation is characteristic for a healthy bone metabolism. Osteoblasts are responsible for bone formation and osteoclasts for bone resorption. In general inflammatory and in particular chronic inflammatory processes influence osteoblast and osteoclast function directly or via indirect mechanisms. Bone metabolism can be influenced by the interaction of cytokines, hormones and growth factors with bone cells. A central factor involved in bone metabolism is the receptor activator of nuclear factor-κB (RANK)/RANK ligand (RANKL)/osteoprotegerin (OPG) system, which is influenced by different inflammatory processes. Usually, (chronic) inflammation results in increased bone loss. The molecular mechanisms and pathophysiological pathways of bone metabolism under the influence of inflammation are summarized in this review.

Cryo-electron microscopy reconstructions of triatoma virus particles: a clue to unravel genome delivery and capsid disassembly.

Triatoma virus (TrV) is a member of the insect virus family Dicistroviridae and consists of a small, non-enveloped capsid that encloses its positive-sense ssRNA genome. Using cryo-transmission electron microscopy and three-dimensional reconstruction techniques combined with fitting of the available crystallographic models, this study analysed the capsids corresponding to mature and several RNA-empty TrV particles. After genome release, the resulting reconstruction of the empty capsids displayed no prominent conformational changes with respect to the full virion capsid. The results showed that RNA delivery led to empty capsids with an apparent overall intact protein shell and suggested that, in a subsequent step, empty capsids disassemble into small symmetrical particles. Contrary to what is observed upon genome release in mammalian picornaviruses, the empty TrV capsid maintained a protein shell thickness and size identical to that in full virions.

Oral cavity colon adenocarcinoma metastases: case report with surgical approach and review of more than 30 years literature.

INTRODUCTION: Metastatic oral tumours are rare, contributing to 1% of all malignant oral cavity tumours. CASE REPORT: We report the case of a 59-year-old man with colon cancer at an advanced disease stage, with progression to the peritoneum and maxillary gingiva. Palliative surgery was indicated to improve the patient's quality of life. DISCUSSION: In a review of the literature, we compiled a list of 27 cases (including the present case) reflecting some 30 years of literature on oral cavity metastatic disease originating in colon cancer. CONCLUSION: Oral cavity metastasis should be taken into account in the differential diagnosis of a synchronous or metachronous oral cavity lesion. The therapeutic goal should include palliative alternatives when necessary. Intraoral reconstruction using local flaps may be a simple and reliable palliative resection option aimed at improving the patient's quality of life.

Is transportation a risk factor for African swine fever transmission in Australia: a review.

African swine fever (ASF) is a viral disease of the pigs that was first described in Africa during the early part of the twentieth century. The disease has periodically occurred outside of Africa, including an ongoing epidemic in Europe and Asia that started in 2007; the disease has never occurred in Australia or New Zealand. Once introduced into a country, spread can occur through direct and indirect routes of transmission. Infected feral pig populations have the potential to act as a long-term reservoir for the virus, making eradication difficult. Just before and throughout the period of clinical signs, ASF virus is shed in oronasal fluids, urine, faeces and blood. This results in contamination of the pig's environment, including flooring, equipment and vehicles. Transportation-related risk factors therefore are likely to play an important role in ASF spread, though evidence thus far has been largely anecdotal. In addition to the existing AUSVETPLAN ASF plan, efforts should be made to improve transportation biosecurity, from the time a pig leaves the farm to its destination. Collection of data that could quantify the capabilities and capacity of Australia to clean and disinfect livestock trucks would help to determine if private and/or public sector investment should be made in this area of biosecurity. No peer-reviewed research was identified that described a specific process for cleaning and disinfecting a livestock truck known to be contaminated with ASF virus, though literature suggests that transportation is an important route of transmission for moving the virus between farms and countries.

The therapeutic use of osmotic minipumps in the severe combined immunodeficiency (SCID) mouse model for rheumatoid arthritis.

OBJECTIVES: The viral gene transfer of interleukin 1 receptor antagonist (IL1ra) and interleukin 10 (IL10) into rheumatoid arthritis (RA) synovial fibroblasts (RASFs) has shown protective effects on cartilage destruction in the severe combined immunodeficiency (SCID) mouse model of RA. Nevertheless, side effects of viral transduction are possible and a number of cytokines or cytokine inhibitors are not available encoded in viral vehicles. As the production of viruses coding for bioactive proteins is cost and time intensive, we established an in vivo long-term release model using osmotic minipumps in the SCID mouse model for RA. METHODS: Isolated RASFs were cultured for four passages and coimplanted together with human cartilage and an Alzet osmotic miniature pump model 2004, containing 200 microl of IL10 and IL1ra for 40 days in SCID mice. Implants were removed after 40 days and evaluated histologically. The actual rates of IL10 and IL1ra in murine serum were measured by ELISA. RESULTS: Release of IL10 and IL1ra by the pumps was effective as both could be measured in significant amounts in the serum of the mice. IL10 and IL1ra release showed protective effects towards the coimplanted cartilage, similar to the adenovirally IL10/IL1ra-transduced RASFs. The mean (SD) invasion scores for the implants with the osmotic pumps were: invasion 0.7 (0.5), degradation 0.5 (0.3) (all parameters significant vs controls, p<0.05). CONCLUSIONS: The results demonstrate that the combination of osmotic pumps with the SCID mouse model for RA can be used as approach for application and evaluation of cartilage-protective molecules. Furthermore, the effect of cartilage-protective cytokines is independent of the type of application.

Differential transcriptome analysis of intraarticular lesional vs intact cartilage reveals new candidate genes in osteoarthritis pathophysiology.

OBJECTIVE: To elucidate disease-specific molecular changes in osteoarthritis (OA) by analyzing the differential gene expression profile of damaged vs intact cartilage areas within the same joint of patients with OA of the knee using a combination of a novel RNA extraction technique and whole-genome oligonucleotide arrays. METHODS: The transcriptome of macroscopically affected vs intact articular cartilage as determined by visual assessment was analyzed using an optimized mill-based total RNA isolation directly from the tissue and high density synthetic oligonucleotide arrays. Articular cartilage samples were obtained from patients with OA of the knee. Expression of differentially regulated genes was validated by real-time quantitative polymerase chain reaction and immunohistochemistry. RESULTS: The amount of RNA obtained by the optimized extraction procedure was at least 1 microg per 500 mg of cartilage and fulfilled the common quality requirements. After hybridization onto HG-U133 Plus 2.0 GeneChips (Affymetrix), 28.6-51.7% of the probe sets on the microarray showed a detectable signal above the signal threshold in the individual samples. A subset of 411 transcripts, which appeared to be differentially expressed, was obtained when applying predefined filtering criteria. Of these, six genes were found to be up-regulated in the affected cartilage of all patients, including insulin-like growth factor binding protein 3 (IGFBP-3), wnt-1-inducible signaling protein 1 (WISP-1), aquaporin 1 (AQP-1), delta/notch-like EGF-repeat containing transmembrane (DNER), decay accelerating factor (DAF), complement factor I (IF). CONCLUSION: The optimized methodical approach reported here not only allows to determine area-specific gene expression profiles of intraindividually different low-RNA containing OA cartilage specimens. In addition, this study also revealed novel genes not yet reported to play a role in the pathophysiology of joint destruction in OA.

Long-term effects of chondrospheres on cartilage lesions in an autologous chondrocyte implantation model as investigated in the SCID mouse model.

Microtraumata often lead to articular cartilage lesions. Due to the bradytrophic character of hyaline cartilage, these lesions are hardly repaired by the organism. Autologous chondrocyte implantation (ACI) was established for restoring isolated structural cartilage defects in knee joints. However, results are not always convincing. Human chondrocytes from patients undergoing total knee arthroplasty were cultured in monolayer followed by condensing single chondrocytes to spheroids (chondrospheres). The integrative capacity of chondrospheres was examined by implanting them into lesions in human articular cartilage specimens and co-implanting them into SCID mice. Mice were sacrificed after 4, 12 and 24 weeks. HE and safranin O staining as well as immunohistochemistry using anti-S100, anti-collagen I and II antibodies were performed and analyzed using semiquantitative scores. Integration of the chondrospheres with the (native) cartilage matrix was analyzed by determining the percentage of adhering surface. With respect to long-term stability, the chondrocytes within chondrospheres showed a typical chondrocytic morphology. Immunohistochemically, a high collagen II production was detected. Over a time period of 24 weeks, an increasing content of collagen type II, glycosaminoglycans and collagenous fibers were found. Importantly, the newly synthesized cartilaginous matrix integrated continuously with the native cartilage lesion border. In conclusion, the presented data demonstrate that chondrospheres are able to restore and conserve their phenotype for at least 24 weeks under in vivo conditions. Moreover, chondrospheres adhere to full-thickness cartilage defects and appear to produce a cartilaginous extracellular matrix which fuses with native cartilage thus generating an autologous cartilage-like repair tissue.