No evidence of Borrelia in cutaneous infiltrates of B-cell lymphomas with a highly sensitive, semi-nested real-time polymerase chain reaction targeting the 5S-23S intergenic spacer region.

BACKGROUND: The role of Borrelia in the development of skin lymphomas has been under discussion for decades. A similar association has been shown for Helicobacter pylori and gastric lymphomas (MALT type). Nevertheless, few molecular studies investigated Borrelia in skin lymphomas and the results are controversial. METHODS: We analysed 46 formalin-fixed, paraffin-embedded skin specimens of clincopathologically confirmed B-cell lymphomas (15 marginal zone lymphomas; 20 follicular lymphomas; three diffuse large B-cell lymphomas; eight secondary cutaneous infiltrates) taken from 36 patients from Northern Germany, an endemic area for Borrelia. Fifteen pseudolymphomatous lesions of cutaneous Borreliosis served as the control. Both groups were examined with a real-time (rt) PCR and a semi-nested PCR targeting the 5S-23S intergenic spacer region (IGS). A multiplex PCR was used to investigate B-cell clonality in all lymphomatous infiltrates (Biomed Primers). RESULTS: With both assays no Borrelia burgdorferi-specific DNA was identified in any of the B-cell lymphomas, while all 15 Borreliosis specimens gave a positive PCR result in the semi-nested PCR protocol, 12 were also positive in the rt PCR (P < 0.01). All B-cell lymphomas showed monoclonal IgH-Rearrangement. Analysis of cutaneous B-cell lymphomas from available studies including ours (n = 334) reveals an odds ratio <1. CONCLUSION: While some previous studies suggested an association between B. burgdorferi and the development of cutaneous B-cell lymphomas in endemic areas, we were unable to confirm this in our patients, despite a highly sensitive Borrelia PCR assay. Our results including meta-analysis of previous studies question the need for antibiotic therapy in patients with cutaneous B-cell lymphomas.

Anaerobic digestion of dried/shredded food waste in a periodic anaerobic baffled reactor.

The objective of the current work is to study the impact of the operational parameters' variation (HRT, OLR and T) on biomethane productivity in a periodic anaerobic baffled reactor (PABR). The feedstock used was a biomass product named food residue biomass (FORBI), which is dried and shredded source-separated household food waste. The PABR is an innovative, high-rate bioreactor. Apart from the hydraulic retention time (HRT) and the organic loading rate (OLR), an important operational parameter is the switching period (T) of the feeding compartment: when T is high, the bioreactor operation is similar to an anaerobic baffled reactor (ABR), while when it is low, the operation approaches that of an upflow anaerobic sludge blanket reactor (UASBR). Nine distinct experimental phases were conducted, during which the operational parameters of the PABR were consecutively modified: the HRT varied from 9 to 2.5 days, T between 2 days and 1 and finally the OLR from 1.24 gCOD/Lbioreactor*d to 8.08 gCOD/Lbioreactor*d. The maximum biomethane yield was 384 LCH4/kgFORBI corresponding to the operation at HRT = 5 d, OLR = 2.14 gCOD/Lbioreactor*d and T = 2 days. Similar efficiency (333 LCH4/kg-FORBI) was achieved at higher OLR (4.53 gCOD/Lbioreactor*d).

Arbuscular mycorrhizal fungus inocula from coastal sand dunes arrest olive cutting growth under salinity stress.

Cultivation of olive trees covers large coastal areas of land in Mediterranean regions, many of them characterized by low soil fertility and exposed to salinity and seasonal drought. In this frame, we developed mixed community inocula of arbuscular mycorrhizal fungi (AMF) derived from the extreme, seasonally arid environments of six Mediterranean sand dunes and evaluated their effects, in the form of community inocula, on rooted semi-woody olive tree cuttings (Olea europaea cv. Koroneiki). The plantlets were grown in the greenhouse for 10 months under 50 mM and 100 mM concentrations of NaCl, successively applied to induce osmotic stress. Inoculation had a positive effect on plant growth and nutrient uptake. However, the three best-performing inocula in early colonization and in plant growth enhancement also resulted in high plant sensitivity to high salinity, which was not observed for the other three inocula. This was expressed by decreased nutrient uptake and drastically lower plant growth, plant photosynthesis, and stomatal conductance (generally an over 50% reduction compared to no salinity application). Amplicon sequencing analysis of the olive plants under salinity stress showed that the AMF communities in the roots were clearly differentiated by inoculation treatment. We could not, however, consistently associate the plant responses observed under high salinity with specific shared AMF community membership or assembly attributes. The observed physiological overreaction to osmotic stress may be an adaptation trait, potentially brought about by host selection coupled to abiotic environmental filtering, in the harsh conditions from which the AMF inocula were derived. The overreaction may, however, be undesirable if conveyed to allochthonous plants at an agronomic level.

Bioelectricity production from fermentable household waste in a dual-chamber microbial fuel cell.

In this study, the use of a dual-chamber microbial fuel cell for the production of bioelectricity from a food residue biomass (FORBI) product was investigated. Food residue biomass was produced by drying and shredding the pre-sorted fermentable fraction of household food waste collected door-to-door in the Municipality of Halandri, Athens, Greece. Different organic loads of food residue biomass expressed as chemical oxygen demand (COD) were examined (0.7, 0.9, 1.4, 2.8, 6 and 14 g COD L-1, respectively). It was observed that an increase of the initial concentration of the final extract resulted in a corresponding increase in the operating time. The microbial fuel cell potential increased from 33.3 mV to 46 mV as the concentration was increased from 0.7 to 14 g COD L-1. The best performance in terms of maximum power density (29.6 mW m-2) corresponding to a current density of 88 mA m-2 was observed for 6 g COD L-1. Setting the external resistance at its optimal value (Rext = 2 kΩ) as determined by polarisation experiments, Pyield drastically increased to 13.7 and 17.3 Joule (g FORBI)-1 in two consecutive cycles. The results demonstrate that readily biodegradable substrates, such as food residue biomass, can be effectively used for enhanced bioelectricity harvesting in a microbial fuel cell.

Vascular endothelial growth factor polymorphisms and clinical outcome in patients with metastatic breast cancer treated with weekly docetaxel.

The aim of the study was to evaluate the association of vascular endothelial growth factor (VEGF) genotypes with treatment efficacy in a phase II trial. This study evaluated weekly docetaxel, as first-line treatment for metastatic breast cancer. Existing data from in vitro and animal model experiments suggest that docetaxel at low doses has anti-angiogenic activity. DNA was extracted from blood samples of 86 patients participating in the trial. Genotyping was performed for selected single-nucleotide polymorphisms (SNPs; VEGF-2578, -1498, -1154, and +936). Moreover, due to the highly polymorphic nature of the studied areas, we were able to analyze additional registered SNPs. All candidate genotypes were evaluated for associations with overall survival (OS), progression-free survival (PFS) and response rate. The VEGF-1154 GG genotype was more frequent in patients not responding to treatment compared with responders (42.9% vs 0.0%, P=0.048). Moreover, the VEGF-2578 AA genotype was associated with longer PFS compared with CC (hazard ratio (HR)=0.40; 95% confidence interval (CI) 0.17-0.98; pairwise P=0.0457). Patients with the VEGF-1190 GG genotype demonstrated shorter PFS compared with those with the alternative genotypes (GA and AA) combined (HR=3.85; 95% CI: 1.20-12.50; P=0.0224). In addition, the VEGF-2551/-2534 homozygous del18bp and VEGF-2430/-2425 homozygous ins1bp genotypes were associated with worse PFS compared with no deletion and no insertion, respectively (HR=2.49; 95% CI: 1.02-6.07; pairwise P=0.0442 and HR=2.57; 95% CI: 1.05-6.27; pairwise P=0.0385, respectively). Furthermore, patients with the VEGF-1498 CC genotype exhibited longer median OS compared with those with the alternatives genotypes (CT and TT) combined (HR=0.27; 95% CI: 0.08-0.89; P=0.0311). In multivariate analysis, the VEGF-2578 AA genotype retained its significance (P=0.0220) for PFS. Our results support the association of specific VEGF genotypes with clinical outcome in patients with metastatic breast cancer treated with a potentially anti-angiogenic regimen, such as weekly docetaxel. However, current results should be validated prospectively in larger cohorts.

A systematic review and meta-analysis of experimental clinical evidence on initial aligning archwires and archwire sequences.

The aim of the study was to assess treatment effects and potential side effects of different archwires used on patients receiving orthodontic therapy. Electronic and manual unrestricted searches were conducted in 19 databases including MEDLINE, Cochrane Library, and Google Scholar until April 2012 to identify randomized controlled trials (RCTs) and quasi-RCTs. After duplicate study selection, data extraction, risk of bias assessment with the Cochrane risk of bias tool, and narrative analysis, mean differences (MDs) with confidence intervals (CIs) of similar studies were pooled using a random-effects model and evaluated with GRADE. A total of 16 RCTs were included assessing different archwire characteristics on 1108 patients. Regarding initial archwires, meta-analysis of two trials found slightly greater irregularity correction with an austenitic-active nickel-titanium (NiTi) compared with an martensitic-stabilized NiTi archwire (corresponding to MD: 1.11 mm, 95% CI: -0.38 to 2.61). Regarding archwire sequences, meta-analysis of two trials found it took patient treated with a sequence of martensitic-active copper-nickel-titanium (CuNiTi) slightly longer to reach the working archwire (MD: 0.54 months, 95% CI: -0.87 to 1.95) compared with a martensitic-stabilized NiTi sequence. However, patients treated with a sequence of martensitic-active CuNiTi archwires reported general greater pain intensity on the Likert scale 4 h and 1 day after placement of each archwire, compared with a martensitic-stabilized NiTi sequence. Although confidence in effect estimates ranged from moderate to high, meta-analyses could be performed only for limited comparisons, while inconsistency might pose a threat to some of them. At this point, there is insufficient data to make recommendations about the majority of initial archwires or for a specific archwire sequence.

Application of the Huang-Hilbert transform and natural time to the analysis of seismic electric signal activities.

The Huang method is applied to Seismic Electric Signal (SES) activities in order to decompose them into their components, named Intrinsic Mode Functions (IMFs). We study which of these components contribute to the basic characteristics of the signal. The Hilbert transform is then applied to the IMFs in order to determine their instantaneous amplitudes. The results are compared with those obtained from the analysis in a new time domain termed natural time, after having subtracted the magnetotelluric background from the original signal. It is shown that these instantaneous amplitudes, when combined with the natural time analysis, can be used for the distinction of SES from artificial noises.

Diet and feeding strategy of blackmouth catshark Galeus melastomus.

Diet and feeding strategy of the blackmouth catshark Galeus melastomus in the deep waters of the eastern Ionian Sea were investigated. Sampling was carried out using experimental bottom longline fishing at depths ranging from 300 to 855 m in summer and autumn 2010. Diet variability with fish size, season, area, sex and depth zone was tested and only season was found to significantly affect the diet of the species. Of the 870 stomachs examined, only 1·4% were empty. Cumulative prey curves showed that the sample sizes were adequate to describe the main prey items of the diet for both seasons. Prey identified belonged primarily to three major groups: fishes, cephalopods and dendrobranchiatan and caridean shrimps. In autumn, the above three major groups were found as principal prey, whereas in summer cephalopods followed by fishes were the principal prey and shrimps were found as secondary prey. SIMPER analysis indicated high dissimilarity between seasons and highlighted that Sepiolidae, Myctophidae, fishes, cephalopods, shrimps and other crustaceans contributed to seasonal differences in the diet of G. melastomus. The prey diversity index was higher in autumn than in summer. A high dietary overlap was observed between the two seasons. Galeus melastomus behaved as an opportunistic predator with a variety of unimportant prey in its diet. Feeding strategy indicated that G. melastomus holds a generalist niche with a considerable specialization at the individual level. This strategy seems to be an adaptation to a food-scarce environment, as typified in deep-water habitats.

Strong host-specific selection and over-dominance characterize arbuscular mycorrhizal fungal root colonizers of coastal sand dune plants of the Mediterranean region.

Sand dunes of the Mediterranean region constitute drought-stressed, low-fertility ecosystems. Arbuscular mycorrhizal fungi (AMF) are regarded as key components of their biota, that contribute to plant host adaptation and fitness. However, AMF community assembly rules in the roots of the psammophilous plants of coastal sand dunes have not been investigated. We studied the root colonizing AMF communities of four characteristic native plants of eastern Mediterranean coastal foredunes, in nine locations in Greece. Host-specificity (plant identity) was the major driver of AMF community assembly in the plant roots, while geographical distance between locations was not related to differences in the AMF communities. Additionally, colonizer AMF communities were characterized by over-dominance of a single operational taxanomic unit (OTUs), which was remarkably host-specific among locations. Wider dissimilarity in AMF communities was observed in small and disturbed (SD) sites compared to large and undisturbed (LU) sites, a trait that may be attributed to relaxed environmental filtering and facilitated AMF dispersal/immigration in SD sites from surrounding habitats. Overall, our results indicate that the assembly of root-colonizing AMF communities in the eastern Mediterranean sand dunes is characterized by strong biotic filtering (host identity), suggesting that co-adaptation processes may be more pronounced than previously proposed, under extreme environmental conditions.

alpha1-Acid glycoprotein production in rat dorsal air pouch in response to inflammatory stimuli, dexamethasone and honey bee venom.

This study shows the rapid and differential production of the 40-43 kDa and the 70-90 kDa alpha1-acid glycoprotein (AGP) fucosylated glycoforms after treatment of the dorsal air pouch with bacterial lipopolysaccharide (LPS), HgCl(2) or Freund's complete adjuvant (FCA). The 40-43 kDa and the 70-90 kDa AGP production is peaked 1-3 h post-LPS treatment. We observed that the responses to LPS and FCA are similar in that both AGP isoforms are induced whereas they differ in that the FCA exhibits a 6 h lag period. The response to HgCl(2,) however, exhibits the specific biphasic induction only of the 40-43 kDa AGP. The serum 40-43 kDa AGP glycoform gradually increases in response to all of the above stimulants and peaks by 24 h post- treatment. The increase of the 70-90 kDa AGP levels in the air pouch occurs in association with the accumulation of polymorphonuclear (PMN) cells while dexamethasone (DEX) increases only the 40-43 kDa AGP production in the absence of PMN accumulation. Macrophage-monocyte lineage cells forming the air pouch lining tissue may potentially be the cells that secrete the 40-43 kDa AGP while polymorphonuclear cells that infiltrate the air pouch secrete the 70-90 kDa AGP. The 40-43 kDa and 70-90 kDa AGP production induced by LPS in the air pouch precedes that of interleukin-1 (IL-1) or interleukin-6 (IL-6) while the 40-43 kDa AGP glycoform potentially increases IL-6 production by air pouch PMN exudate cells. These significant differences suggest a local pro-inflammatory role of AGP. Honeybee venom suppressed arthritis development and exhibited differential local or systemic regulation of AGP in serum vs. air pouch exudate or synovial fluid. This study with the air pouch model of facsimile synovium tissue suggests that local alpha1-acid glycoprotein (AGP) production may contribute to pro-inflammatory and anti-inflammatory activities during the local acute phase response or during chronic inflammatory stress as in arthritis.

Left ventricular function in elite rowers in relation to training-induced structural myocardial adaptation.

To examine left ventricular (LV) function in elite young athletes in relation to structural adaptation to prolonged intense training. Conventional echocardiography and tissue Doppler imaging (TDI) were performed in 15 elite rowers and 12 sedentary matched controls. Rowers had increased LV mass index, septal (12 vs 10 mm, P<0.005) and posterior wall thicknesses (12 vs 9 mm, P<0.001) and increased relative wall thickness. Septal and lateral systolic velocities were enhanced in rowers (septal S(m)=8.5 vs 6.3 cm/s, P<0.001; lateral S(m)=11.4 vs 8.0 cm/s, P<0.005), representing a 35% and 42% increase, respectively. Similarly, septal and lateral early diastolic velocities were enhanced (septal E(m)=12.1 vs 9.5 cm/s, P<0.01; lateral E(m)=16.6 vs 11.6 cm/s, P<0.001), representing a 27% and 43% increase, respectively. Systolic and early diastolic TDI velocities of the lateral wall showed a positive correlation (r=0.65, P<0.01) in athletes indicating a parallel improvement of systolic and diastolic function, while LV stiffness was decreased [(E/E(m))/(LV end-diastolic diameter)=1.13 vs 1.57, P<0.005). Both systolic and diastolic LV function were improved in elite rowers, despite a pattern of concentric hypertrophy.

Programmes of measures of the marine strategy framework directive: Are they contributing to achieving good environmental status in the Mediterranean?

The Marine Strategy Framework Directive (MSFD) is the piece of legislation with which the EU aims to achieve Good Environmental Status (GES) of its seas. This Directive requires Member States (MSs) to set up and implement Programmes of Measures (PoMs) to achieve this goal. This paper presents a catalogue as well as the analysis of the 535 measures proposed by the eight Mediterranean EU MSs to combat pollution and non-indigenous species and contribute to the GES of the Mediterranean Sea. The results reiterate the need for better coherence and coordination between MSs, also with non-EU countries in the preparation of the next iteration of PoMs. Only 5% of the measures applied have an effect beyond MSs´ marine waters jurisdiction, and a fifth of them lack linkages with any ecosystem component. The results point to gaps, while desired goals and recommendations will help policy makers to implement the and conceptualise measures.

Long-term changes in the structure of benthic communities: Revisiting a sampling transect in Crete after 24 years.

A bathymetric transect in the north coast of Crete first studied in 1989, was revisited 24 years later. Identical sampling design, season, techniques and protocols were followed in both studies in order to minimize bias in the long-term comparisons. This comprehensive macrofaunal dataset (4 stations, 2 sampling seasons, 7 replicates in each study) revealed changes in benthic diversity and community composition between the sampling periods. The recorded changes were higher at the stations located close to the coastal zone. In addition, while benthic communities showed lower total abundance during the recent sampling period, species abundances were more evenly distributed indicating that some species dominated the historical communities. In spite of these changes, the ecological status remained above the threshold values for good ecological status. The results indicated that changes in the benthic community seem to have been driven by local anthropogenic factors and natural variability rather than by large-scale factors such as nutrients influxes in the entire Mediterranean Basin.

Bioremediation of a soil contaminated by lindane utilizing the fungus Ganoderma australe via response surface methodology.

Mixtures of a sandy soil and wheat straw were doped with the organochlorine insecticide lindane in glass tubes and were inoculated with the polypore fungus, Ganoderma australe. An evaluation of bioremediation process effectiveness was searched and five parameters identified for the solid-state system. Fungi growth is a function of temperature and requires moisture for a proper colonization. These microorganisms need inorganic nutrients such nitrogen and phosphorus to support cell growth and it is also appropriate to know the range of concentration and toxicity of the used insecticide. Thus, an orthogonal central composite design (CCD) of experiments was used to construct second order response surfaces. Five design factors, namely temperature, moisture, straw, lindane content and nitrogen content and seven optimization parameters (responses), namely lag time, propagation velocity, biomass growth rate, biodegradation rate, biodegradation/biomass, biomass/propagation and biomass content were analyzed. The optima of the responses of the adequate models were found to be the following: propagation velocity 4.25mm/day, biomass growth rate 408mg/day, biodegradation/biomass 56.9microg/g, biomass/propagation 250mg/mm and fungal biomass content in solid mixture 260mg/cm(3). The most important response for bioremediation purposes is biodegradation/biomass which is maximized at the factors levels: temperature 17.3 degrees C, moisture 58%, straw content 45%, lindane content 13ppm and nitrogen content 8.2ppm.

Biodegradation of lindane by Pleurotus ostreatus via central composite design.

The degradation of lindane was studied in liquid-agitated cultures using a commercial strain of the fungus Pleurotus ostreatus as the biodegrading organism. The biodegradation was accomplished with the action of extracellular oxidative enzymes, produced by the fungus to decompose woody substrates. Enzyme activities of manganese peroxidase and laccase were measured in a liquid mineral medium. An orthogonal Central Composite Design of experiments was used to construct second-order response surfaces with the fungus growth, final pH and the lindane biodegradation as optimization parameters. The initial lindane concentration, the nitrogen content, the incubation time and the temperature were used as design factors. Optimal conditions found for all these parameters will be used for the continuation of this project aiming at the bioremediation of contaminated sites with persistent organic pollutants such as lindane.

An increase in creatine kinase secondary to acute pancreatitis: a case report.

Creatine kinase (CK) activity is found in high concentrations in skeletal muscle, cardiac muscle and brain. Here, we describe a 64-year-old woman with acute pancreatitis and elevated serum CK activity. This association is extraordinarily rare. In particular, laboratory findings which were found to be abnormal were serum CK 4.150 U/l (peaked 1 day after admission) with the CK-MB fraction being less than 5%, lactate dehydrogenase (LDH) 424 U/l, serum lipase 1.265 U/I and serum amylase 1.105 U/l. Some data regarding the phenomenon of acute pancreatitis and elevated serum CK activity are given.

Determination of synthetic food colorants in fish products by an HPLC-DAD method.

Reliable methods for quantification of synthetic water-soluble colors in complex food matrices are currently not available. The present work describes the development and validation of an improved protocol for the analysis of synthetic food colorants in complex food matrices presenting high protein and/or fat content. The method developed employs an extraction stage, followed by a subsequent sonification, centrifugation and concentration step. A final clean up via SPE on polyamide cartridges was also employed. The isolated colorants were separated and analyzed by an RP-HPLC/DAD system. High and consistent recoveries (min. 81%) and low RSDs (max. 6%) were achieved for all studied colorants. The issue of high fat content matrices was also addressed showing the need for an additional defatting step in the procedure. Overall, the protocol presented shows high precision and accuracy of detection and can provide the basis for future development of similar methods in other complex food matrices.

Detection of human papillomaviruses in squamous cell carcinomas of the lung.

The aim of this study was to evaluate the possible association of human papillomaviruses (HPV) with the development of squamous cell lung carcinomas (SqCLCs). Tissue material from 52 cases of SqCLCs were studied, and the data were evaluated according to the degree of differentiation, HPV presence and type. Analysis was performed by polymerase chain reaction (PCR) method using consensus primers, and the results were confirmed by subsequent Southern blot hybridization. Overall, the results showed 69% positivity (n=32). Forty-one cases were examined for the presence of specific HPV types (6/11 and 16/18) by hybridization of the PCR products with 32P-labelled probes. HPV 6/11 types were detected in 6 of the 29 positive cases (20.6%). HPV 16/18 types were the most prevalent types, and were detected in 11/29 cases (37.9%: 4/10 of well-differentiated cases, 6/25 of moderately and 1/6 of poorly differentiated carcinomas). Our results confirm the possibility that HPV might play a role in the development of SqCLCs and suggest a possible relation of high-risk HPV16/18 types to tumour differentiation.

Chemical composition of essential oils from needles and twigs of balkan pine (Pinus peuce grisebach) grown in Northern Greece.

The composition of essential oils from twigs and needles of Balkan pine (Pinus peuce Gris.) grown in northern Greece was investigated. The compounds were identified by using GC-MS analysis. The twig oil was rich in alpha-pinene (7.38%), beta-pinene (12.46%), beta-phellandrene (26.93%), beta-caryophyllene (4.48%), and citronellol (12.48%), and the needle oil was rich in alpha-pinene (23.07%), camphene (5.52%), beta-pinene (22.00%), beta-phellandrene (6.78%), bornyl acetate (9.76%), beta-caryophyllene (3.05%), and citronellol (13.42%). The mean oil yield was 2.85% for twigs and 0. 57% for needles.