Long-term opioid therapy of non-cancer pain : Prevalence and predictors of hospitalization in the event of possible misuse.

BACKGROUND: One major concern of long-term opioid therapy (LTOT) for chronic noncancer pain (CNCP) is the risk of abuse of prescribed opioids. OBJECTIVE: To examine the prevalence and predictors of opioid use-related hospitalizations and potential abuse of prescribed opioids by persons with LTOT for CNCP in a sample representative of the German statutory health insurance companies. METHODS: Retrospective cross-sectional study in 2014. Anonymized German health claims database, including 4,028,618 insured individuals of 69 German statutory health insurances. Univariate logistic regression models to evaluate demographic and medical characteristics associated with hospital stays and a diagnosis of mental and behavioral disorders due to alcohol, opioids, tranquilizers, multiple substances and intoxications by narcotic agents in insured individuals with CNCP receiving LTOT. RESULTS: The prevalence of LTOT for CNCP was 0.8%; 9.9% of these insured individuals received high-dose LTOT (≥120 morphine equivalent mg/day). The 1­year prevalence of hospital stays with a diagnosis of mental and behavioral disorders due to alcohol, opioids, tranquilizers, multiple substances and intoxications by narcotic agents was 1.75% of persons with LTOT. These diagnoses were strongly associated with prescriptions of tranquilizers (odds ratio [OR] 3.63; 95% confidence interval [CI] 3.03; 4.36) and moderately associated with diagnosis of depression (OR 2.52; 95% CI 2.12; 3.00) and slightly associated with diagnosis of somatoform pain disorder (OR 1.89; 95% CI 1.56; 2.28) and high-dose LTOT (OR 1.81; 95% CI 1.44; 2.27). DISCUSSION: The study is in line with the recommendations of the German national guidelines on long-term opioid therapy of chronic non-cancer pain (LONTS) to avoid concomitant prescription of tranquilizers for CNCP and to carefully select and monitor patients with depression and somatoform pain disorder.

Regional Spatial-Temporal Spread of Citrus Huanglongbing Is Affected by Rain in Florida.

Citrus huanglongbing (HLB), associated with 'Candidatus Liberibacter asiaticus' (Las), disseminated by Asian citrus psyllid (ACP), has devastated citrus in Florida since 2005. Data on HLB occurrence were stored in databases (2005 to 2012). Cumulative HLB-positive citrus blocks were subjected to kernel density analysis and kriging. Relative disease incidence per county was calculated by dividing HLB numbers by relative tree numbers and maximum incidence. Spatiotemporal HLB distributions were correlated with weather. Relative HLB incidence correlated positively with rainfall. The focus expansion rate was 1626 m month-1, similar to that in Brazil. Relative HLB incidence in counties with primarily large groves increased at a lower rate (0.24 year-1) than in counties with smaller groves in hotspot areas (0.67 year-1), confirming reports that large-scale HLB management may slow epidemic progress.

[Long-term opioid therapy of non-cancer pain : Prevalence and predictors of hospitalization in the event of possible misuse]. / Langzeitopioidtherapie von nichttumorbedingten Schmerzen : Prävalenz und Prädiktoren von Krankenhausaufenthalten bei möglicher missbräuchlicher Verwendung.

BACKGROUND: One major concern of long-term opioid therapy (LTOT) for chronic noncancer pain (CNCP) is the risk of abuse of prescribed opioids. OBJECTIVE: To examine the prevalence and predictors of opioid use-related hospitalizations and potential abuse of prescribed opioids by persons with LTOT for CNCP in a sample representative of the German statutory health insurance companies. METHODS: Retrospective cross-sectional study in 2014. Anonymized German health claims database, including 4,028,618 insured individuals of 69 German statutory health insurances. Univariate logistic regression models to evaluate demographic and medical characteristics associated with hospital stays and a diagnosis of mental and behavioral disorders due to alcohol, opioids, tranquilizers, multiple substances and intoxications by narcotic agents in insured individuals with CNCP receiving LTOT. RESULTS: The prevalence of LTOT for CNCP was 0.8%; 9.9% of these insured individuals received high-dose LTOT (≥120 morphine equivalent mg/day). The 1­year prevalence of hospital stays with a diagnosis of mental and behavioral disorders due to alcohol, opioids, tranquilizers, multiple substances and intoxications by narcotic agents was 1.75% of persons with LTOT. These diagnoses were strongly associated with prescriptions of tranquilizers (odds ratio [OR] 3.63; 95% confidence interval [CI] 3.03; 4.36) and moderately associated with diagnosis of depression (OR 2.52; 95% CI 2.12; 3.00) and slightly associated with diagnosis of somatoform pain disorder (OR 1.89; 95% CI 1.56; 2.28) and high-dose LTOT (OR 1.81; 95% CI 1.44; 2.27). DISCUSSION: The study is in line with the recommendations of the German national guidelines on long-term opioid therapy of chronic non-cancer pain (LONTS) to avoid concomitant prescription of tranquilizers for CNCP and to carefully select and monitor patients with depression and somatoform pain disorder.

Enhanced vascular biocompatibility of decellularized xeno-/allogeneic matrices in a rodent model.

Glutaraldehyde preservation is the gold standard for cardiovascular biological prosthesis. However, secondary calcifications and the absence of tissue growth remain major limitations. Our study assessed in vitro and in vivo the biocompatibility of human (fascia lata, pericardium) and porcine tissues (pericardium, peritoneum) treated with a physicochemical procedure for decellularization and non-conventional pathogens inactivation. Biopsies were performed before and after treatment to assess decellularization (HE/Dapi staining/DNA quantification/MHC I/alpha gal immunostaining) and mechanical integrity. Forty-five rats received an abdominal aortic patch of native cryopreserved tissues (n = 20), treated tissues (n = 20) or glutaraldehyde-preserved bovine pericardium (GBP, control, n = 5). Grafts were explanted at 4 weeks and processed for HE/von Kossa staining and immunohistochemistries for lymphocytes (CD3)/macrophages (CD68) histomorphometry. 95% of decellularization was obtained for all tissues except for fascia lata (75%). Mechanical properties were slightly altered. In the in vivo model, a significant increase of CD3 and CD68 infiltrations was found in native and control implants in comparison with decellularized tissues (p < 0.05). Calcifications were found in 3 controls. Decellularized tissues were recolonized. GBP showed the most inflammatory response. This physicochemical treatment improves the biocompatibility of selected xeno/allogeneic tissues in comparison with their respective native cryopreserved tissues and with GBP. Incomplete decellularization is associated with a significantly higher inflammatory response. Our treatment is a promising tool in the field of tissue decellularization and tissue banking.

Impact of perioperative chemotherapy on survival in patients with advanced primary urethral cancer: results of the international collaboration on primary urethral carcinoma.

BACKGROUND: To investigate the impact of perioperative chemo(radio)therapy in advanced primary urethral carcinoma (PUC). PATIENTS AND METHODS: A series of 124 patients (86 men, 38 women) were diagnosed with and underwent surgery for PUC in 10 referral centers between 1993 and 2012. Kaplan-Meier analysis with log-rank testing was used to investigate the impact of perioperative chemo(radio)therapy on overall survival (OS). The median follow-up was 21 months (mean: 32 months; interquartile range: 5-48). RESULTS: Neoadjuvant chemotherapy (NAC), neoadjuvant chemoradiotherapy (N-CRT) plus adjuvant chemotherapy (ACH), and ACH was delivered in 12 (31%), 6 (15%) and 21 (54%) of these patients, respectively. Receipt of NAC/N-CRT was associated with clinically node-positive disease (cN+; P = 0.033) and lower utilization of cystectomy at surgery (P = 0.015). The objective response rate to NAC and N-CRT was 25% and 33%, respectively. The 3-year OS for patients with objective response to neoadjuvant treatment (complete/partial response) was 100% and 58.3% for those with stable or progressive disease (P = 0.30). Of the 26 patients staged ≥cT3 and/or cN+ disease, 16 (62%) received perioperative chemo(radio)therapy and 10 upfront surgery without perioperative chemotherapy (38%). The 3-year OS for this locally advanced subset of patients (≥cT3 and/or cN+) who received NAC (N = 5), N-CRT (N = 3), surgery-only (N = 10) and surgery plus ACH (N = 8) was 100%, 100%, 50% and 20%, respectively (P = 0.016). Among these 26 patients, receipt of neoadjuvant treatment was significantly associated with improved 3-year relapse-free survival (RFS) (P = 0.022) and OS (P = 0.022). Proximal tumor location correlated with inferior 3-year RFS and OS (P = 0.056/0.005). CONCLUSION: In this series, patients who received NAC/N-CRT for cT3 and/or cN+ PUC appeared to demonstrate improved survival compared with those who underwent upfront surgery with or without ACH.

Molecular Detection of the Laurel Wilt Fungus, Raffaelea lauricola.

The laurel wilt disease fungus, Raffaelea lauricola, is killing redbay trees, spreading rapidly in the U.S. southeastern coastal plain forest, and posing a serious threat to the avocado industry in Florida. A molecular tool is urgently required to facilitate detection of this pathogen. The 5' region of the large ribosomal RNA (28S) gene is highly variable among Raffaelea spp. and ideal for this purpose but amplification of this sequence from R. lauricola has been difficult. Different amplification conditions were tested and a high-fidelity polymerase chain reaction (PCR) procedure utilizing a dNTP mix containing 7-deaza-dGTP was found to reliably amplify 28S sequences from R. lauricola. Sequencing the amplified products or cloned inserts also turned out to be difficult and required using a custom-blended sequencing mix containing 1 M betaine, 5% dimethyl sulfoxide, and dGTP-BigDye v3.1. Three GC-rich stem and loop or cruciform secondary structures were discovered, which may have interfered with amplification. This improved protocol made it possible to partially characterize the internal transcribed spacers sequence from R. lauricola, which also has interfering secondary structures. A TaqMan real-time PCR assay was designed using the species-specific 28S sequences and this allowed detection of R. lauricola from wood tissues or cultures. Wood tissues from symptomatic redbay, avocado, and sassafras trees in Florida were screened using this TaqMan assay and several were found to test positive for R. lauricola. Results were further confirmed by performing Koch's postulates for avocado specimens collected from commercial grooves.

First Report of Rose rosette virus Associated with Rose Rosette Disease Infecting Knockout Roses in Florida.

Roses are one of the most popular flowering shrubs in the United States, with a total wholesale value of US$194 million. Among the major states, Florida is the fourth largest producer of roses with a total value exceeding US$20 million (4). In Florida, the roses have become especially popular in recent years with the introduction of Knock Out and other shrub roses. Virus-like symptoms including witches'-broom, excessive thorns, abnormal red discoloration of shoots and foliages, distorted leaves, and deformed buds and flowers were initially observed on Knock Out roses in a commercial nursery in Quincy, FL, in November 2013. Fifteen plants out of ~250,000 plants showed these characteristic symptoms. Total RNA extracts (RNeasy Plant Mini Kit, Qiagen, Valencia, CA) from eight symptomatic and two non-symptomatic rose samples were subjected to reverse-transcription (RT) assays using SuperScript III Reverse transcriptase (Invitrogen, Life Technologies, NY) and random hexamer primers. The cDNA synthesized was then subjected to PCR assay using Platinum Taq DNA polymerase (Invitrogen, Life Technologies) and using Rose rosette virus (RRV) specific primers RRV-F and RRV-R (1), targeting the core region of the RNA1 genome of the virus. The RT-PCR assays using the specific primers produced amplicons of 375 bp, only in the symptomatic leaf samples. The obtained amplicons were PCR purified and sequenced directly (GenBank Accession Nos. KF990370 to KF990377). BLAST analysis of these sequences revealed a higher identity of 99% with the RRV (HQ871942) in the NCBI database. Pairwise comparison of the eight RRV sequences exhibited 99 to 100% identity among themselves. These results revealed the association of RRV with the symptomatic rose plants. Eight symptomatic and two non-symptomatic rose plant samples were tested for RRV using blot hybridization assay, utilizing a digoxigenin-labeled DNA probe of 511 bp, targeting the RNA1 genome of the RRV. All eight symptomatic rose plants showed a positive reaction to the RRV-specific probes, confirming the presence of RRV in the samples, while the non-symptomatic and the buffer control did not produce any reactions. Even though the virus is reported to spread by an eriophyid mite Phyllocoptes fructiphilus, thorough examination of the infected samples showed absence of the vector. The samples were also tested using RT-PCR for the presence of Rose cryptic virus (RCV) and Blackberry chlorotic ringspot virus (BCRV) using specific primers (2,3). The samples tested negative for the RCV and BCRV. This is the first report of occurrence of RRV on rose in Florida. Considering the economic importance of the rose plants and the highly destructive nature of RRV, this report underscores the need for immediate effective quarantine and management of the virus for protecting the economically important rose industry in Florida. References: (1) A. G. Laney et al. J. Gen. Virol. 92:1727, 2011. (2) S. Sabanadzovic and N. Abou Ghanem-Sabanadzovic. J. Plant Pathol. 90:287, 2008. (3) I. E. Tzanetakis et al. Plant Pathol. 55:568, 2006. (4) USDA. 2007 Census of Agriculture 3:25, Washington, DC, 2010.

Vascular study of decellularized porcine long bones: Characterization of a tissue engineering model.

INTRODUCTION: Massive bone allografts enable the reconstruction of critical bone defects in numerous conditions (e.g. tumoral, infection or trauma). Unfortunately, their biological integration remains insufficient and the reconstruction may suffer from several postoperative complications. Perfusion-decellularization emerges as a tissue engineering potential solution to enhance osseointegration. Therefore, an intrinsic vascular study of this novel tissue engineering tool becomes essential to understand its efficacy and applicability. MATERIAL AND METHODS: 32 porcine long bones (humeri and femurs) were used to assess the quality of their vascular network prior and after undergoing a perfusion-decellularization protocol. 12 paired bones were used to assess the vascular matrix prior (N = 6) and after our protocol (N = 6) by immunohistochemistry. Collagen IV, Von Willebrand factor and CD31 were targeted then quantified. The medullary macroscopic vascular network was evaluated with 12 bones: 6 were decellularized and the other 6 were, as control, not treated. All 12 underwent a contrast-agent injection through the nutrient artery prior an angio CT-scan acquisition. The images were processed and the length of medullary vessels filled with contrast agent were measured on angiographic cT images obtained in control and decellularized bones by 4 independent observers to evaluate the vascular network preservation. The microscopic cortical vascular network was evaluated on 8 bones: 4 control and 4 decellularized. After injection of gelatinous fluorochrome mixture (calcein green), non-decalcified fluoroscopic microscopy was performed in order to assess the perfusion quality of cortical vascular lacunae. RESULTS: The continuity of the microscopic vascular network was assessed with Collagen IV immunohistochemistry (p-value = 0.805) while the decellularization quality was observed through CD31 and Von Willebrand factor immunohistochemistry (p-values <0.001). The macroscopic vascular network was severely impaired after perfusion-decellularization; nutrient arteries were still patent but the amount of medullary vascular channels measured was significantly higher in the control group compared to the decellularized group (p-value <0.001). On average, the observers show good agreement on these results, except in the decellularized group where more inter-observer discrepancies were observed. The microscopic vascular network was observed with green fluoroscopic signal in almost every canals and lacunae of the bone cortices, in three different bone locations (proximal metaphysis, diaphysis and distal metaphysis). CONCLUSION: Despite the aggressiveness of the decellularization protocol on medullary vessels, total porcine long bones decellularized by perfusion retain an acellular cortical microvascular network. By injection through the intact nutrient arteries, this latter vascular network can still be used as a total bone infusion access for bone tissue engineering in order to enhance massive bone allografts prior implantation.

Blisters - an unusual effect during radiotherapy.

The skin reaction to radiation is regularly monitored in order to detect enhanced radiosensitivity of the patient, unexpected interactions (e.g. with drugs) or any inadvertent overdosage. It is important to distinguish secondary disease from radiation reaction to provide adequate treatment and to avoid unnecessary discontinuation of radiotherapy. A case of bullous eruption or blisters during radiotherapy of the breast is presented. Differential diagnoses bullous pemphigoid, pemphigus vulgaris, and bullous impetigo are discussed and treatment described.

[Bariatric surgery - significance, risks, long term consequences]. / Adipositaschirurgie- Stellenwert, Risiken und Folgen.

In recent years the number of bariatric surgery has markedly increased in industrial nations. Surgery provides a more rapid decrease of body weight than conservative approach. However a long term conservative follow up therapy is mandatory to stabilize reduced weight. Due to increasing knowledge from long term follow up of surgically treated obese patients there is a growing body of evidence that frequently there is necessity of reoperations and of substitution both of trace elementsand of minerals or vitamins due to their hampered enteral resorption. Additionally therapy of surgery induced endocrine alterations not seldom is necessary.These insights are of outstanding importance because meanwhile an enlargement of the indications for bariatric surgery as a therapeutic option for metabolic disorders is being discussed. This review refers to the recent internationally published papers concerning consequences of bariatric surgery.

First Report of Pseudomonas cichorii Causing Leaf Spot of Stevia Detected in Florida.

During a routine nursery inspection in Hernando County, Florida in July 2011, leaf spot symptoms were observed on the popular sugar substitute stevia (Stevia rebaudiana). Spots were roughly circular to irregular in shape, variable in size, and dark brown with a yellow halo. White to cream-colored, circular, convex, gram-negative bacterial colonies were isolated on nutrient agar and identified as Pseudomonas cichorii based on the LOPAT scheme (2). DNA from four individual colonies inoculated in nutrient broth was extracted using DNeasy columns (Qiagen Inc., Valencia, CA). The PCR-amplified product from four different genes, 16S rRNA (1.5 Kbp), gyrB (0.9 Kbp), rpoB (1.2 Kbp), and rpoD (0.7 Kbp), was sequenced (GenBank Accession Nos. JQ994483, JQ994484, JQ994485, and JQ994486). Nucleotide and translated amino acid sequences for each gene were compared to the nucleotide and protein databases, respectively. The best matches were always with P. cichorii with nucleotide identities ranging from 98 to 99% and amino acid identities from 99 to 100%. Four healthy stevia plants were spray inoculated with 20 ml of a 108 CFU ml-1 suspension prepared from a 24-hour-old culture, of which two were sprayed with carborundum (silicon carbide) immediately prior to inoculation. Two additional plants were sprayed with carborundum only or sterile tap water only and served as healthy controls. Symptoms began to develop 4 days after inoculation. Spots originated at the tips or edges of the leaves and enlarged over time. Enlarging lesions progressed to encompass the entire leaf, accompanied by water soaking, curling, and necrosis. Blighting typically spread down to the stem and caused dieback. The pathogen was successfully reisolated from the lesions and produced identical LOPAT scheme results. Based on the information collected, it is believed that this is the first confirmed report of bacterial leaf spot caused P. cichorii on stevia worldwide. This find is significant due to the rising popularity of stevia cultivation for its sweetness and medicinal properties (1). To our knowledge, stevia is not currently being grown commercially in Florida; however, there is commercial acreage being developed elsewhere in the United States. References: (1) D. Patil et al. Asian J. Pharm. Clin. Res. 5:1, 2011. (2) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd Edition. The American Phytopathological Society. St. Paul, MN, 2001.

First Report of Guignardia citricarpa Associated with Citrus Black Spot on Sweet Orange (Citrus sinensis) in North America.

In March 2010, citrus black spot symptoms were observed on sweet orange trees in a grove near Immokalee, FL. Symptoms observed on fruit included hard spot, cracked spot, and early virulent spot. Hard spot lesions were up to 5 mm, depressed with a chocolate margin and a necrotic, tan center, often with black pycnidia (140 to 200 µm) present. Cracked spot lesions were large (15 mm), dark brown, with diffuse margins and raised cracks. In some cases, hard spots formed in the center of lesions. Early virulent spot lesions were small (up to 7 mm long), bright red, irregular, indented, and often with many pycnidia. In addition, small (2 to 3 mm), elliptical, reddish brown leaf lesions with depressed tan centers were observed on some trees with symptomatic fruit. Chlorotic halos appeared as they aged. Most leaves had single lesions, occasionally up to four per leaf. Tissue pieces from hard spots and early virulent spots were placed aseptically on potato dextrose agar (PDA), oatmeal agar, or carrot agar and incubated with 12 h of light and dark at 24°C. Cultures that grew colonies within a week were discarded. Fourteen single-spore cultures were obtained from the isolates that grew slower than the Guignardia mangiferae reference cultures, although pycnidia formed more rapidly in the G. mangiferae cultures (1). No sexual structures were observed. Cultures on half-PDA were black and cordlike with irregular margins with numerous pycnidia, often bearing white cirrhi after 14 days. Conidia (7.1 to 7.8 × 10.3 to 11.8 µm) were hyaline, aseptate, multiguttulate, ovoid with a flattened base surrounded by a hyaline matrix (0.4 to 0.6 µm) and a hyaline appendage on the rounded apex, corresponding to published descriptions of G. citricarpa (anomorph Phyllosticta citricarpa) (1). A yellow pigment was seen in oatmeal agar surrounding G. citricarpa, but not G. mangiferae colonies as previously reported (1,2). DNA was extracted from lesions and cultures and amplified with species-specific primers (2). DNA was also extracted from G. mangiferae and healthy citrus fruit. The G. citricarpa-specific primers produced a 300-bp band from fruit lesions and pure cultures. G. mangiferae-specific primers produced 290-bp bands with DNA from G. mangiferae cultures. The internally transcribed spacer (ITS) of the rRNA gene, translation-elongation factor (TEF), and actin gene regions were sequenced from G. citricarpa isolates and deposited in GenBank. These sequences had 100% homology with G. citricarpa ITS sequences from South Africa and Brazil, 100% homology with TEF, and 99% homology with actin of a Brazilian isolate. Pathogenicity tests with G. citricarpa were not done because the organism infects immature fruit and has an incubation period of at least 6 months (3). In addition, quarantine restrictions limit work with the organism outside a contained facility. To our knowledge, this is the first report of black spot in North America. The initial infested area was ~57 km2. The disease is of great importance to the Florida citrus industry because it causes serious blemishes and significant yield reduction, especially on the most commonly grown 'Valencia' sweet orange. Also, the presence of the disease in Florida may affect market access because G. citricarpa is considered a quarantine pathogen by the United States and internationally. References: (1) R. P. Baayen et al. Phytopathology 92:464, 2002. (2) N. A. Peres et al. Plant Dis. 91:525, 2007 (3) R. F. Reis et al. Fitopath Bras. 31:29, 2006.

First Report of a 16SrIV-D Phytoplasma Associated with Texas Phoenix Palm Decline on Pigmy Date Palm (Phoenix roebelenii) in Florida.

The pigmy date palm (Phoenix roebelenii O'Brien) is used as an ornamental in Florida and is popular and lucrative within the nursery trade. Severe decline of several pigmy date palms was observed at a residence in Hillsborough County, FL. Several other palm species, including P. canariensis (Canary Island date palm), P. sylvestris (wild date palm), P. dactylifera (date palm), Syagrus romanzoffiana (queen palm), and Sabal palmetto (cabbage palm), in Florida are known to be affected by Texas Phoenix palm decline (TPPD), a disease associated with a phytoplasma subgroup strain 16SrIV-D (2,3). Moreover, the location of the affected pigmy date palms was in the proximity of many other diseased cabbage palms that were identified in previous surveys and subsequently rogued. Genomic DNA was extracted from 100 mg of ground-up palm trunk tissues containing phloem cells with a DNeasy Plant Mini kit column (QIAGEN Inc., Valencia, CA) from four specimens. A high-fidelity PCR (Hf-PCR) procedure was used in preference to standard PCR because it was 10,000 to 100,000 times more sensitive (1,4). The Hf-PCR (50 µl) utilized two DNA polymerases; Taq (five units) and ACCUZYME (one unit), 350 µM dNTP, a buffer (50 mM Tris pH 9.2, 16 mM ammonium sulfate, and 1.75 mM magnesium chloride), a higher concentration of primers (200 pM) (2,3), and palm DNA templates (>10 ng) or no DNA negative control. Hf-PCR was performed using three linked profiles: (i) 94°C (2 min) (1 cycle); (ii) 94°C (10 s), 50°C (30 s) for P1m/P7 or 57°C for LY16Sf/LY16Sr and 68°C (2 min) (10 cycles); and (iii) 94°C (10 s), 50°C (30 s) for P1m/P7 or 57°C for LY16Sf/LY16Sr and 68°C (2 min plus 20 s added for every consecutive cycle) (20 cycles) (1). The genomic DNA extracted from P. roebelenii specimens was used as template for amplification by Hf-PCR. Expected 1.8- and 1.4-kb DNA bands for each primer combination were readily amplified. The Hf-PCR products were sequenced (GenBank Accession No. JF791816) and a BLAST search revealed a 100% similarity with a phytoplasma subgroup strain 16SrIV-D (EF042899 and AF434989), which is known to cause severe palm decline (TPPD) in other hosts (2,3). To our knowledge, this is the first report of TPPD from P. roebelenii, and therefore, expands the host range of this pathogen. In areas where TPPD is present, the landscape industry may need to identify alternative nonhost palm species or resistant varieties for disease management. References: (1) W. M. Barnes. Proc. Natl. Acad. Sci. USA 91:2216, 1994. (2) N. A. Harrison et al. Plant Dis. 86:676, 2002. (3) N. A. Harrison et al. Ann. Appl. Biol. 153:85, 2008. (4) A. Jeyaprakash and M. A. Hoy. Insect Mol. Biol. 9:393, 2000.

Laurel Wilt, Caused by Raffaelea lauricola, is Confirmed in Miami-Dade County, Center of Florida's Commercial Avocado Production.

Laurel wilt, caused by Raffaelea lauricola, threatens native and nonnative species in the Lauraceae in the southeastern United States, including the important commercial crop, avocado, Persea americana (2,4). Although the pathogen's vector, Xyleborus glabratus, was detected in Miami-Dade County, FL in January 2010, laurel wilt had not been reported (4). In February 2011, symptoms of the disease were observed on native swampbay, P. palustris, in Miami-Dade County (25°72'N, 80°48'W). Externally, foliage was brown, necrotic, and did not abscise; internally, sapwood was streaked with dark gray-to-bluish discoloration; and, in dead trees, holes of natal galleries of the vector from which columns of frass were attached were evident. On a semiselective medium for R. lauricola, a fungus with the pathogen's phenotype was isolated from symptomatic sapwood. Colonies were slow growing, light cream in color, with dendritic, closely appressed mycelium and often a slimy surface. A representative strain of the fungus was further identified with PCR primers for diagnostic small subunit (SSU) rDNA (1) and its SSU sequence (100% match, GenBank Accession No. JN578863). In each of two experiments, plants of 'Simmonds' avocado, the most important cultivar in Florida, were inoculated with three strains of the fungus, as described previously (3). Symptoms of laurel wilt developed in all inoculated plants and the fungus was recovered from each. After aerial and further ground surveys, additional symptomatic swampbay trees, some of which had defoliated, were detected in the vicinity of the original site. Since swampbay defoliates only a year or more after symptoms develop (4), the 2010 detection of X. glabratus may have coincided with an undetected presence of the disease. As of July 2011, a 6-km-diameter disease focus was evident in the area, the southernmost edge of which is 5 km from the nearest commercial avocado orchard. In August 2011, a dooryard avocado tree immediately north of the above focus was affected by laurel wilt, and an SSU sequence confirmed the involvement of R. lauricola (GenBank Accession No. JN613280). The outbreak of laurel wilt in Miami-Dade County represents a 150 km southerly jump in the distribution of this disease in the United States ( http://www.fs.fed.us/r8/foresthealth/laurelwilt/dist_map.shtml ) and is the first time this disease has been found in close proximity to Florida's primary commercial avocado production area. Approximately 98% of the state's commercial avocados, worth nearly $54 million per year, are produced in Miami-Dade County. Since effective fungicidal and insecticidal measures have not been developed for large, fruit-bearing trees, mitigation efforts will focus on the rapid identification and destruction of infected trees (3,4). References: (1) T. J. Dreaden et al. Phytopathology 98:S48, 2008. (2) S. W. Fraedrich et al. Plant Dis. 92:215, 2008. (3) R. C. Ploetz et al. Plant Dis. 95:977, 2011. (4) R. C. Ploetz et al. Recovery Plan for Laurel Wilt of Avocado. National Plant Disease Recovery System, USDA, ARS, 2011.

Complication rates of percutaneous brachial artery puncture: effect of live ultrasound guidance.

PURPOSE: The current literature on the use of brachial artery access is controversial. Some studies found increased puncture site complications. Others found no higher complication rates than in patients with femoral or radial access. The purpose of this study was to determine the impact of ultrasound (US)-guidance on access site complications. MATERIALS AND METHODS: This is a single-center retrospective study of all consecutive patients with brachial arterial access for interventional procedures. Complications were classified into minor complications (conservative treatment only) and major complications (requiring surgical intervention). The brachial artery was cannulated in the antecubital fossa under US-guidance. After the intervention, manual compression or closure devices, both followed by a compression bandage for 3 h, either achieved hemostasis. RESULTS: Seventy-five procedures in seventy-one patients were performed in the study period using brachial access. Access was successful in all cases (100%). Procedures in different vascular territories were performed: neurovascular (10/13.5%), upper extremity (32/43.2%), visceral (20/27.0%), and lower extremity (12/16.3%). Sheath size ranged from 3.2F to 8F (mean: 5F). Closure devices were used in 17 cases (22.7%). In total, six complications were observed (8.0%), four minor complications (5.3%, mostly puncture site hematomas), and two major complications, that needed surgical treatment (2.7%). No brachial artery thrombosis or upper extremity ischemia occurred. CONCLUSION: Exclusive use of US-guidance resulted in a low risk of brachial artery access site complications in our study compared to the literature. US-guidance has been proven to reduce the risk of access site complications in several studies in femoral access. In addition, brachial artery access yields a high technical success rate and requires no additional injection of spasmolytic medication. Sheath size was the single significant predictor for complications.

Golden-Angle Radial Sparse Parallel (GRASP) MRI differentiates head & neck paragangliomas from schwannomas.

PURPOSE: The aim of this study was to demonstrate the value of DCE MRI with high spatiotemporal resolution (GRASP) for differentiating paragangliomas and schwannomas in the head and neck. METHODS: In a retrospective PACS search of in total 410 patients who had undergone head & neck GRASP-MRI, we identified 6 patients with biopsy proven cervical paragangliomas (n = 3) and schwannomas (n = 3). Conventional MRI features were evaluated, lesion size was determined. Postprocessing in 4D-GRASP datasets was performed (1) based on reconstructions with a temporal resolution (Tres) of 4.1 s, qualitative time-intensity curve classification and semiquantitative parameter (Tpeak, PH, ERmax and Slopemax) analysis, and (2) voxel-based mapping and qualitative and semiquantitative perfusion modeling based on reconstructions with a Tres of 1.6 s. Additionally, GRASP perfusion analysis was performed in another set of 5 patients with presumed cervical paragangliomas (n = 3) and schwannomas (n = 2) based on conventional imaging criteria and was correlated with conventional imaging findings. Due to the small sample size, both groups were compared qualitatively. RESULTS: In the time intensity curve classification of 4D GRASP reconstructions (Tres 4.1 s), biopsy proven paragangliomas were consistently characterized by a type-III rapid inflow wash-out pattern, compared to a type-I inflow pattern in the schwannoma group. In both temporal resolutions, semiquantitative analysis of time intensity curves demonstrated rapid wash-in, wash-out, and higher peak signal intensities in paragangliomas compared to schwannomas. In 5 presumed (non-biopsy-proven) paragangliomas and schwannomas, time intensity curves improved diagnostic certainty. CONCLUSIONS: Visual time intensity curve classification and semi-quantitative analysis of GRASP-MRI were, in this small retrospective series, sufficient to differentiate cervical paragangliomas from schwannomas. Utilization of this technique may further improve diagnostic confidence in lesions lacking conventional imaging features.

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.

Role of the netrin system of repellent factors on synovial fibroblasts in rheumatoid arthritis and osteoarthritis.

Changes in the expression of repellent factors, i.e., Netrins and their receptors, may be responsible for the invasive behavior of the synovial tissue cells in patients with rheumatoid arthritis (RA) and osteoarthritis (OA). This study was carried out to analyze the expression of Netrins and their receptors in synovial cells of patients with RA, OA, and control subjects without synovial inflammation. Quantitative RT-PCR was performed to measure the expression of Netrin-1, -3, -4, Neogenin, DCC, UNC5A-D. The influence of Netrin-1 on synovial fibroblasts (SF) was analyzed by determining proliferation, migration, and their ability to organize collagen. SF expressed all repellent factors of the Netrin family. When comparing SF of healthy donors to patients with RA and OA, a stronger expression of UNC5B (4 fold) and UNC5C (769 fold) in RA and OA was found, whereas expression of the other molecules revealed no significant differences. Treating the SF-cells with recombinant Netrin-1 resulted in inhibition of migration of RA- and OA-SFs whereas control cells were not affected. The stronger expression of UNC5B and UNC5C receptors might contribute to the disordered phenotype of RA- and OA-SFs. Addition of Netrin-1 reduces the migratory ability of SFs, potentially by repulsion, as seen in neuronal cells in embryonic development. Due to its function, Netrin-1 may constitute a novel target in the treatment of OA and RA.

Gallium nitride electrodes for membrane-based electrochemical biosensors.

We report on the deposition of planar lipid bilayers (supported membranes) on gallium nitride (GaN) electrodes for potential applications as membrane-based biosensors. The kinetics of the lipid membrane formation upon vesicle fusion were monitored by simultaneous measurements of resistance and capacitance of the membrane using AC impedance spectroscopy in the frequency range between 50 mHz and 50 kHz. We could identify a two-step process of membrane spreading and self-healing. Despite its relatively low resistance, the membrane can be modeled by a parallel combination of an ideal resistor and capacitor, indicating that the membrane efficiently blocks the diffusion of ions.

Lower density of synovial nerve fibres positive for calcitonin gene-related peptide relative to substance P in rheumatoid arthritis but not in osteoarthritis.

OBJECTIVES: Sensory nerve fibres (NFs) contain two major neuropeptides, substance P (SP) and calcitonin gene-related peptide (CGRP). The pro-inflammatory role of SP is known, while CGRP has anti-inflammatory activities by inhibiting T helper type 1 cytokines, TNF secretion and leucocyte proliferation. We demonstrated the increase of SP-positive NFs in RA as compared with OA. This study investigated the density of CGRP-positive NFs relative to SP-positive NFs or sympathetic NFs in synovial tissue of patients with RA and OA. METHODS: By immunofluorescent staining of synovial tissue of 25 patients with RA and 35 patients with OA, NFs positive for CGRP, SP and tyrosine hydroxylase (sympathetic NFs) were quantified. RESULTS: Density of CGRP-positive NFs was higher in OA than in RA, and density of SP-positive NFs tended to be higher in RA. In RA patients, comparison of CGRP-positive and SP-positive NFs in the same synovial tissue demonstrated less CGRP-positive than SP-positive NFs. The ratio of CGRP-positive NFs to SP-positive NFs was lower in RA as compared with OA. In OA, but not in RA, density of CGRP-positive NFs positively correlated with density of sympathetic NFs, which is much lower in RA patients. CONCLUSION: The preponderance of SP-positive NFs over CGRP-positive NFs or sympathetic NFs most probably supports the pro-inflammatory process in patients with RA. The reasons for the loss of CGRP in sensory NFs are not known.