GIP receptor suppresses PAC1receptor-mediated neuronal differentiation via formation of a receptor heterocomplex.

Pituitary adenylate cyclase-activating peptide (PACAP) receptor (PAC1R) is a class B Gprotein-coupled receptor (GPCR) that is widely expressed in the human body and is involved in neuronal differentiation. As class B GPCRs are known to form heterocomplexes with family members, we hypothesized that PAC1R mediates neuronal differentiation through interaction with a class B GPCR. We used the BRET assay to identify potential interactions between PAC1R and 11 class B GPCRs. Gastric inhibitory polypeptide receptor (GIPR) and secretin receptor were identified as putative binding partners of PAC1R. The effect of heterocomplex formation by PAC1R on receptor activation was evaluated with the cyclic (c)AMP, luciferase reporter, and calcium signaling assays; and the effects on receptor internalization and subcellular localization were examined by confocal microscopy. The results suggested he PAC1R/GIPR heterocomplex suppressed signaling events downstream of PAC1R, including cAMP production, serum response element and calcium signaling, and ß-arrestin recruitment. Protein-protein interaction was analyzed in silico, and induction of neuronal differentiation by the PAC1R heterocomplex was assessed in SH-SY5Y neuronal cells by measure the morphological changes and marker genes expression by real-time quantitative PCR and western blot. Over-expression of GIPR suppressed PACAP/PAC1R-mediated neuronal differentiation and the differentiation markers expression in SH-SY5Y cells. GIPR regulates neuronal differentiation through heterocomplex formation with PAC1R.

Biosorptive interaction of alkaline modified Dialium guineense seed powders with ciprofloxacin in contaminated solution: central composite, kinetics, isotherm, thermodynamics, and desorption.

This work evaluated the use of Dialium guineense seed waste (DGS) and its sodium hydroxide modified form (NH-DGS) as biosorbent for ciprofloxacin (CPF) from synthetic solution as well as the desorption potentials. Central composite design (CCD) was applied for optimization of the alkaline treated biosorbent by response surface methodology using design expert. Both biosorbents were characterized by FTIR, SEM, EDX, and BET analysis. The CCD showed NaOH concentration of 0.46 M and temperature of 96 °C to be effective for optimized modification of NH-DGS. Optimum removal of CPF was obtained at pH 6.0, contact time 120 min, temperature 300 K, and dosage of 0.1 g. The Freundlich model gave the best fit compared to the other isotherms tested with R2 values >0.97951. NH-DGS exhibited a maximum uptake capacity of 120.34 mg/g higher than some reported adsorbents for CPF. The pseudo-second-order model was suitable in the fitting of the kinetic data. A non-spontaneous process was obtained for CPF biosorption on DGS which became spontaneous after alkaline treatment. Over 84% desorption of CPF was achieved on both biosorbents using 0.3 M HCl which envisaged the use of NH-DGS as an efficient material for treatment of waters contaminated with CPF.

Anthropogenic impact on water chemistry and benthic macroinvertebrate associated changes in a southern Nigeria stream.

The Ogba River in southern Nigeria is an important water resource for its riparian communities. This study evaluates impact of anthropogenic influences on the Ogba River using water chemistry and macroinvertebrate data sets obtained over a period of 6 months between January and June 2012. Four stations, stations 1-4, characterised by various human activities were chosen along the river. Organic wastes from domestic and industrial sources were the major point sources of pollutants. Station 2 where the municipal wastewater drains into the river had elevated values of flow velocity, BOD5, sulphate, phosphate, nitrate and sodium. Based on the canonical correspondence analysis (CCA), 5-day biochemical oxygen demand (BOD5), sulphate, nitrate and phosphate were the main factors that help to shape the macroinvertebrate assemblage structure of the Ogba River. Macroinvertebrates clustered strongly by stations than by seasons indicating that water quality differences between the stations were responsible for the observed differences in the biotic assemblage. The preponderance of naidid oligochaetes, baetid nymphs and certain tolerant dipteran taxa including chironomids and ceratopogonids at all four stations was an indication that the entire water body was stressed. The odonates were the single most abundant taxa; their dominance could be attributed to the vegetative nature of the stream, favouring odonate colonisation. Overall, the responses of macroinvertebrates to stress were reflected by the different assemblage structures recorded at the four study stations. Substrate and microhabitat obliteration and poor water quality appeared to be the factors responsible for the observed assemblage structure in the Ogba River.

Longitudinal cortical split sign as a potential diagnostic feature for cortical osteitis.

Septic cortical osteitis is a rare but distinct type of bone infection that is characterized as a hematogenously seeded infection predominantly or exclusively limited to the cortex. Diagnosis is difficult and often delayed. Combination of clinical and laboratory findings together with the typical radiological findings consisting of vertically orientated cortical osteolysis, the 'cortical split sign' and the predominantly cortical disruption at the periosteal side of the cortex may lead to the correct diagnosis.

Radiation-induced sarcoma: analysis of 46 cases.

A retrospective analysis was performed of 46 cases of sarcoma treated in our institution between 1989 and 2007 that occurred in a previously irradiated area. Eight male and 38 female patients had received radiotherapy, mainly for breast cancer and genitourinary tumours. The interval between irradiation and the diagnosis of sarcoma ranged from 1 to 54 years (median 15 y). The most common clinical findings were a mass, pain and skin dislocation. Angiosarcoma and sarcoma non-otherwise-specified were the most common histological types. Surgical resection was performed in 34 patients (74%) and 5-year survival was 45% when a radical resection was obtained. No 5-year survival was noticed after non-radical resection and in the absence of surgery. Stage and location of the sarcoma were other prognostic factors. Overall 5-year survival was 27% for the whole group.

Ewing's sarcoma: imaging features.

AIM: To define an imaging prototype of Ewing's sarcoma (ES). MATERIALS AND METHODS: Sixty-four patients with a histopathologically and/or genetically proven diagnosis of ES were analyzed for clinical parameters (age, gender and location), radiographic and CT appearance (distribution, matrix, margins, periosteal reaction, articular extension, cortical reaction and the presence of a pathologic fracture). Size, local extension, signal intensity, degree and pattern of enhancement, and the presence of skip metastases were evaluated on MRI. Distant metastases were recorded on bone scintigraphy and chest CT scan. RESULTS: Patient's age ranged between 7-67 (mean 17.9). Male/female ratio was 2.4/1. Location in the pelvis was most frequent (31%), followed by the femur (20%) and tibia (11%). Most tumors were mixed lytic-sclerotic (75%), and purely lytic in 25%. Plain films and CT scan showed a spiculated periosteal reaction in 50%. A Codman's triangle was seen in 27%. Articular extension was difficult to assess on radiographs. Cortical permeation and destruction is seen in respectively 31 and 42%, whereas cortical thickening is seen in 20%. Pathologic fracture occurred in 7.8%. MRI showed a large mass, with a soft tissue component of more than 50% in 67%. Degree and pattern of enhancement pattern was variable. Signal intensity on T1- and T2-WI was non-specific. Joint involvement was seen in 23%. Isolated involvement of the soft tissue (extraskeletal ES) was seen in 1.5%. Skip metastases at initial presentation were present at initial presentation in 14% and distant metastases in 22%. CONCLUSIONS: ES occurs in young patients. On radiographs/CT, 37.5% are located in the axial skeleton and 62.5% in the peripheral skeleton. ES is mostly mixed sclerotic-lytic. A spiculated periosteal reaction is most frequent. The most characteristic finding on MRI is the presence of a large soft tissue mass.

Imaging of malignant bonetumors.

Diagnostic imaging plays a pivotal role in the initial detection, characterization, pre-operative assessment and long term follow-up of malignant bone tumors. The purpose of this brief review is to discuss the specific role of the different imaging modalities in the diagnostic work-up of malignant bone tumors. The imaging features, with emphasis on standard radiography, allowing differentiation, of malignant bone tumors, will be highlighted.

Expression of angiogenic VEGF-A (soluble isoforms 121, 165) and lymphangiogenic VEGF-C in colorectal cancers with micro-satellite instability.

BACKGROUND AND OBJECTIVES: Colorectal cancers (CRC) with high-level micro-satellite instability (MSI-H) show reduced metastatic potential and better prognosis compared to stage-matched stable (MSS) cancers. Angiogenesis/lymphangiogenesis, central to tumour growth and spread, is mediated by vascular endothelial growth factor (VEGF) cytokines, but little is known of their relationship to MSI. METHODS: In this study, 67 sporadic CRC with identified MSI status, and 8 samples of normal colon were analysed for VEGF-A soluble isoforms (VEGF-121/VEGF-165) and VEGF-C gene transcription (by RT-PCR and scanning densitometry), and blood vessel density (BVD; measuring angiogenesis) and VEGF-C protein expression (measuring lymphangiogenesis). RESULTS: Compared to normal colon, VEGF-165 transcription was reduced (P < 0.05), but VEGF-121 transcription was higher in MSS (P < 0.06) and MSI-L (P < 0.01) cancers (but similar in MSI-H). VEGF-165 transcription was unrelated to MSI, but VEGF-121 transcription was elevated in MSI-L (P < 0.06). There was a weak inverse correlation with VEGF-121 transcription and Dukes stage (P < 0.09), and with BVD and MSI (P < 0.09). With a singular di-nucleotide loci mutation (MSI-L), VEGF-121 (P < 0.03) and VEGF-C (P < 0.04) transcription was elevated. CONCLUSIONS: MSI-H cancers have reduced angiogenic/lymphangiogenic potential, and transcription of VEGF-121 may be important in early growth and spread of CRC. Elevated VEGF-121 and VEGF-C transcription with singular di-nucleotide mutations may aid in the identification of distinct MSI-L cancers.

Multiple vertebral fluid-fluid levels.

We present a case of multiple vertebral metastases, with multiple fluid-fluid levels, from a moderately to poorly differentiated carcinoma of unknown origin. We suggest that fluid-fluid levels in multiple vertebral lesions are highly suggestive of bone metastases.

Non perforated septum supra-patellaris mimicking a soft tissue tumour.

Plicae synoviales of the knee are well known to present as anterior knee pain but can also cause blocking, clicking, and swelling of the knee mimicking pathology such as meniscal tears or other internal derangements, and even mono-arthritis. Embryologically, the knee is divided into a medial and lateral compartment and a supra-patellar pouch by thin membranes of synovium. Incomplete involution of these membranes gives rise to a plica. When there is no involution at all, the membrane is called a septum. There are three frequent types of plicae/septa: a suprapatellar, a medial-patellar, and an infra-patellar type. We discuss on a patient with a non perforated septum supra-patellaris which resulted in a swelling above the knee, mistaken for a soft tissue tumour.

Selective arterial embolization in the treatment of an aneurysmal bone cyst of the pelvis.

This case report deals with an aggressive aneurysmal bone cyst of the pelvis in a 20-year-old man causing considerable destruction of the left ilium and the anterior column of the acetabulum, with extension into the pelvis. The treatment of large pelvic aneurysmal bone cysts is challenging because of local destruction of adjacent structures and because of the risk of severe intraoperative bleeding. Therefore selective preoperative embolization may be a valuable tool in the management of aneurysmal bone cysts.

Medical image based, preformed titanium membranes for bone reconstructions: design study and first clinical.

The currently used intralesional or marginal surgical treatment of a bone tumour in the extremities shows some shortcomings in providing a restoration of the mechanical strength of the bone and the containment of the used filling materials. The use of a medical image based, preformed and custom-made titanium membrane screwed onto the periosteal side of the bone is introduced. This study looks in detail into the design process and the biomechanical evaluation of such a membrane. The buckling strength of the membrane, the strength at the perforation holes and the strength of the screw-bone fixation are tested experimentally. The two latter experiments are performed with different screw types. From the performed tests it appears that a titanium membrane without a wave pattern, of 0.3 mm thickness, fixed to the bone with seven trabecular bone screws (4 mm diameter and 28 mm length) is capable of carrying the anticipated mechanical loads on the reconstructed tibia. The medical image based design methodology and the first clinical application of such a preformed and custom-made titanium membrane are reported and discussed. The feasibility of preformed titanium membranes for bone reconstruction in tumour surgery is demonstrated.

Clinical impact of molecular and cytogenetic findings in synovial sarcoma.

Synovial sarcoma is an aggressive soft-tissue tumor that accounts for up to 10% of soft-tissue sarcomas. Cytogenetically, synovial sarcoma is characterized by the t(X;18)(p11;q11), found in more than 95% of the tumors. This translocation results in rearrangements of the SYT gene in 18q11 and one of the SSX1, SSX2, or SSX4 genes in Xp11, creating a SYT/SSX1, SYT/SSX2, or SYT/SSX4 chimeric gene. It has been shown that patients with SYT/SSX1 fusion genes have a shorter metastasis-free survival than do patients with SYT/SSX2. Previous studies have also suggested that clonal evolution may be associated with disease progression. In the present study, RT-PCR analysis showed that all 64 examined synovial sarcomas from 54 patients had SYT-SSX chimeric genes. SYT/SSX1 was found in 40 tumors from 33 patients, SYT/SSX2 in 23 tumors from 20 patients, and SYT/SSX4 in one case. Two patients had variant SYT/SSX2 transcripts, with 57 bp and 141 bp inserts, respectively, between the known SYT and SSX2 sequences. Patients with tumors with SYT/SSX1 fusions had a higher risk of developing metastases compared to those with SYT/SSX2 fusions (P = 0.01). The reciprocal transcripts SSX1/SYT and SSX2/SYT were detected using nested PCR in 11 of the 40 samples with SYT/SSX1 and 5 of the 23 samples with SYT/SSX2, respectively. Among 20 blood samples, SYT/SSX1 and SYT/SSX2 were detected in one sample each. The t(X;18), or variants thereof, was found cytogenetically in all patients but three. Among 32 primary tumors, the t(X;18) or a variant translocation was the sole anomaly in 10. In contrast, of the seven metastatic lesions that were investigated prior to radiotherapy, only one had a t(X;18) as the sole anomaly; all other tumors displayed complex karyotypes. Cytogenetic complexity in primary tumors was, however, not associated with the development of metastases. Tumors with SYT/SSX2 less often (4/12 vs. 7/15) showed complex karyotypes than did tumors with SYT/SSX1, but the difference was not significant. Combining cytogenetic complexity and transcript data, we found that the subgroup of patients with tumors showing simple karyotypes and SYT/SSX2 fusion had the best clinical outcome (2/8 patients developed metastases), and those with tumors showing complex karyotypes together with SYT/SSX1 fusion the worst (6/7 patients developed metastases). This corresponded to 5-year metastasis-free survival rates of 0.58 and 0.0, respectively (P = 0.02).

Metastatic fractures of the tibia.

Pathologic fractures of the tibia due to a metastasis are rare. The treatment of an established fracture is sometimes conservative, but more often surgical. The purpose of the surgical procedure is to improve the quality of life and the ambulatory status, to relieve pain and to facilitate activities of daily living and nursing care. Four cases of operatively managed metastatic fractures of the tibia are presented with emphasis on the surgical technique. The scarce literature on metastatic tibial fractures in reviewed. The operative technique to be used does not only depend on the location of the tumor but also on the primary tumor, the response to adjuvant therapy and the life expectancy. For metastatic shaft fractures an intramedullary nail, sometimes augmented with bone cement, is preferred. For distal or proximal fractures a compound osteosynthesis with plates and screws offers a good solution. In the epiphyseal and metaphyseal region of the tibia an amputation or a tumor prosthesis is the procedure of choice.

Candidal vertebral osteomyelitis: report of 6 patients, and a review.

The incidence of deep-seated candidal infection is increasing, but candidal vertebral osteomyelitis is still rare. We describe 6 patients recently treated in our hospital. Conservative treatment failed in all. We reviewed the literature and identified 59 additional cases of candidal vertebral osteomyelitis. Candidemia was documented in 61.5% of them. The interval between the diagnosis of candidemia and the onset of symptoms of vertebral osteomyelitis varied widely, from days to >1 year. In patients without documented candidemia, there was a similar interval between the occurrence of risk factors for candidemia (present in 72% of the patients) and the onset of symptoms of vertebral osteomyelitis. Clinical, laboratory, and radiological findings are not specific for candidal spondylodiskitis. Final diagnosis is determined by means of culture of a biopsy specimen from the infected vertebra or disk. Treatment consisted of prolonged antifungal treatment, and it often included surgery. On the basis of our experience (for all 6 patients, initial conservative treatment with only antifungals failed), we recommend consideration of early surgical debridement in combination with prolonged antifungal therapy.

Calcifying/ossifying synovial sarcoma shows t(X;18) with SSX2 involvement and mitochondrial calcifications.

AIMS: Synovial sarcoma with extensive calcification and ossification is a rare variant, the ultrastructural, cytogenetic and molecular analysis of which has not been reported previously. METHODS AND RESULTS: A large mass in the shoulder of a 20-year-old male patient led to a deformity of the chest wall, thus supporting the hypothesis that this is a slowly growing variant of synovial sarcoma. Nevertheless, the patient developed metastatic lung disease 7 months after resection. On histology, the monophasic spindle cell proliferation was in several areas obscured by the massive calcification and ossification. Immunohistochemistry showed keratin, epithelial membrane antigen, vimentin and CD99 expression. The cytogenetic analysis revealed a single t(X;18)(p11.2; q11.2), typical for synovial sarcoma. Additional fluorescence in-situ hybridization revealed SSX2 involvement. At the ultrastructural level, prominent needle-shaped intramitochondrial crystals were present, both in the cytoplasm and in the extracellular matrix. CONCLUSION: The presence of the t(X;18) with SSX2 involvement definitively characterizes this tumour as a variant of synovial sarcoma. In addition, the needle-like mitochondrial calcifications give a possible clue to the pathogenesis of the extensive metaplastic ossification and calcification.

Recurrent chromosome aberrations in fibrous dysplasia of the bone: a report of the CHAMP study group. CHromosomes And MorPhology.

The nosologic status of fibrous dysplasia (FD), a well-known and relatively common bone lesion, is controversial. Information collected by the CHromosomes And MorPhology (CHAMP) study group on published and unpublished cases of fibrous dysplasia shows the presence of clonal chromosome changes in at least a proportion of these lesions. The chromosome aberrations found in FD lesions have been quite variable and have included both structural and numerical changes. Two of the three cases investigated at the study group had trisomy 2 as the sole acquired anomaly. Combined with previously published data, +2 and rearrangements involving chromosome band 12p13 have each been detected in 3 of 8 cases with abnormal karyotype of 11 in which chromosomal analysis has been performed, suggesting that FD is a neoplastic lesion rather than a "dysplastic" process, as has been generally believed and as implied by its very name.

Cytogenetic-morphologic correlations in aneurysmal bone cyst, giant cell tumor of bone and combined lesions. A report from the CHAMP study group.

Aneurysmal bone cyst and giant cell tumor of bone are relatively rare bone tumors that sometimes coexist. We examined the karyotypes of 3 aneurysmal bone cysts, 12 giant cell tumors, and 3 combined lesions. All aneurysmal bone cysts showed involvement of chromosome segments 17p11-13 and/or 16q22. In addition, in 1 of the 3 giant cell tumors with secondary aneurysmal bone cyst, both chromosome bands were rearranged as well, albeit not in a balanced translocation. Seven out of 12 giant cell tumors were characterized by telomeric associations. One giant cell tumor showed a dup(16)(q13q22), suggesting the presence of a (minor) secondary aneurysmal bone cyst component, despite the absence of histological proof. Our results, combined with literature data further substantiate that segments 16q22 and 17p11-13 are nonrandomly involved in at least some aneurysmal bone cysts, irrespective of subtype (primary, secondary, intra/extraosseous, solid or classic). These findings strongly suggest that some aneurysmal bone cysts are true neoplasms. In addition, telomeric associations are the most frequent chromosomal aberrations in giant cell tumor of bone, the significance of which remains elusive. In combined giant cell tumor/aneurysmal bone cyst each component seems to retain its own karyotypic abnormality.