Flow Cytometry Analysis of Neural Differentiation Markers Expression in Human Glioblastomas May Predict Their Response to Chemotherapy

Glioblastoma multiforme (GBM) represents an extremely chemoresistant tumour type. Here, authors analysed the immunophenotype of GBM tumours by flow cytometry and correlated the immunophenotypic characteristics with sensitivity to chemotherapy. The expression of selected neural and non-neural differentiation markers including A2B5, CD34, CD45, CD56, CD117, CD133, EGFR, GFAP, Her-2/neu, LIFR, nestin, NGFR, Pgp and vimentin was analysed by flow cytometry in eleven GBM (WHO gr.IV) patients. The sensitivity of tumour cells to a panel of chemotherapeutic agents was tested by the MTT assay. All tumours were positive for A2B5, CD56, nestin and vimentin. CD133, EGFR, LIFR, NGFR and Pgp were expressed only by minor tumour cell subpopulations. CD34, CD45, CD117, GFAP and Her-2/neu were constantly negative. Direct correlations were found between the immunophenotypic markers and chemosensitivity: A2B5 vs lomustine (r² = 0.642, P = 0.033), CD56 vs cisplatin (r² = 0.745, P = 0.013), %Pgp⁺ vs vincristine (r² = 0.846, P = 0.008), and %NGFR⁺ vs daunorubicine (r² = 0.672, P = 0.047) and topotecan (r² = 0.792, P = 0.011). In contrast, inverse correlations were observed between: EGFR vs paclitaxel (r² = ?0.676, P = 0.046), CD133 vs dacarbazine (r² = ?0.636, P = 0.048) and LIFR vs daunorubicine (r² = ?0.878, P = 0.004). Finally, significant associations were also found among sensitivities to different chemotherapeutic agents and among different immunophenotypic markers. In conclusion, histopathologically identical GBM tumours displayed a marked immunophenotypic heterogeneity. The expression of A2B5, CD56, NGFR and Pgp appeared to be associated with chemoresistance whereas CD133, EGFR and LIFR expression was characteristic of chemosensitive tumours. We suggest that flow cytometric imunophenotypic analysis of GBM may predict chemoresponsiveness and help to identify patients who could potentially benefit from chemotherapy.     

The influence of exogenous carbohydrate provision and pre-exercise alkalosis on the heat shock protein response to prolonged interval cycling

The aim of this study was to observe the intracellular heat shock protein 72 (HSP72) and heme oxygenase-1 (HSP32) response to prolonged interval cycling following the ingestion of carbohydrates (CHO) and sodium bicarbonate (NaHCO₃). Six recreationally active males (mean ± SD; age 23.2 ± 2.9 years, height 179.5 ± 5.5 cm, body mass 76.5 ± 6.8 kg, and peak power output 315 ± 36 W) volunteered to complete a 90 min interval cycling exercise on four occasions. The trials were completed in a random and blinded manner following ingestion of either: placebo and an artificial sweetener (P–P), NaHCO₃ and sweetener (B–P), placebo and CHO (P–CHO), and NaHCO₃ and CHO (B–CHO). Both HSP72 and HSP32 were significantly increased in monocytes and lymphocytes from 45 min post-exercise (p ≤ 0.039), with strong relationships between both cell types (HSP72, r = 0.83; HSP32, r = 0.89). Exogenous CHO had no influence on either HSP72 or HSP32, but the ingestion of NaHCO₃ significantly attenuated HSP32 in monocytes and lymphocytes (p ≤ 0.042). In conclusion, the intracellular stress protein response to 90 min interval exercise is closely related in monocytes and lymphocytes, and HSP32 appears to be attenuated with a pre-exercise alkalosis.     

Alterations in serum selenium levels and their relation to troponin I in acute myocardial infarction

Selenium (Se) is an essential trace element with antioxidant function. The aim of the present study was to estimate the alterations of Se serum level during the acute phase of myocardial infarction and its relation to biomarkers of myocardial necrosis. Serum Se levels were measured at admission and after 24 h in 60 consecutive patients with acute coronary syndrome (both with and without ST elevation). Troponin I (TnI) was assessed at admission and then twice daily for 3 days; patients with normal levels were excluded. Fifty-five patients with acute MI (positive TnI) were included into the analysis. During the first day of hospitalization, patients received standard therapy, including acetylsalicylic acid, clopidogrel, and heparin or enoxaparin; all underwent urgent coronary angiography and percutaneous intervention, when appropriate. Mean Se levels at baseline and 24 h later were comparable (67.1 ± 2.1 vs. 67.2 ± 1.8 μg/L, ns). Linear regression has shown significant correlation between baseline Se levels and peak TnI (y = 3.4x ? 116, r ² = 0.13, P = 0.008). Positive correlation was found also between the peak TnI and the difference from baseline to 24 h (y = 2.2x + 115, r ² = 0.08, P = 0.04). Moreover, close negative correlation was observed between baseline Se levels and the difference from baseline to 24 h (y = ?0.9x + 62.7, r ² = 0.55, P<0.001). Our results have shown marked individual changes in Se levels during the acute phase of MI as well as correlation between Se levels and peak TnI. These results suggest that alterations in serum Se may be related to the extent of myocardial infarction.     

Juvenile idiopathic arthritis patients and their skeletal status: possible role of vitamin D receptor gene polymorphism

We evaluated bone mineralization and metabolism changes related to vitamin D receptor (VDR) polymorphic genotypes in children with juvenile idiopathic arthritis. One hundred and ninety eight children (82 boys and 116 girls) were included in our study. Bone mineral density (BMD) was measured by lumbar spine DXA. Osteocalcin, CTX, parathyroid hormone, total and ionized calcium, inorganic phosphate, total alkaline phosphatase activity was utilized for assessment of bone metabolism. Molecular testing: TaqI (rs731236) and Cdx2 (rs11568820) polymorphisms of VDR were detected by RFLP. No differences in TaqI and Cdx2 haplotypes, genotypes and alleles distribution related with normal and low BMD (Zscore <?2SD) were found. Children with low linear growth (<10th percentile) had more allele T-contained genotypes of TagI VDR (p = 0.037), compare with medium or high linear growth children. Children with high linear growth (>90th percentile) had the highest frequency of allele A-contained genotypes (GA+AA) of Cdx2 VDR (p = 0.009). Girls with TT TaqI VDR, who never been treated by glucocorticoides had lower BMD-Zscore than C allele carriers (TT = ?0.94SD [IQR: ?2.1;?0.5], TC+CC = ?0.62SD [IQR: ?1.26;0.39], p = 0.03). Girls with Tanner I with TT had higher total and ionized Ca level than carriers of C allele (Ca: TT = 2.43 ± 0.15 mmol/l, TC+CC = 2.28 ± 0.2 mmol/l, p = 0.024; Ca²⁺: TT = 1.15 ± 0.08 mmol/l, TC+CC = 1.06 ± 0.13 mmol/l, p = 0.026). Presence of TT genotype negatively correlated with BMD-Zscore (r = ?0.28, p = 0.04), and positively with frequency of LBMD (r = 0.3, p = 0.037). Boy with GG Cdx2 genotype had lower total Ca (GG = 2.3 ± 0.17 mmol/l, GA+AA = 2.43 ± 0.17 mmol/l, p = 0.004) compare with carriers of A allele. Pubertal boys (Tanner IV–V) with GG had higher CTX (GG = 1.75 ± 0.11 ng/ml, GA+AA = 1.06 ± 0.07 ng/ml, p = 0.04. TT genotype of TaqI and GG genotype of Cdx2 VDR is a negative factor impact bone mineralization metabolism and linear growth.     

Alterations in erythrocyte membrane fluidity and Na+/K+-ATPase activity in chronic alcoholics

Ethanol disorders biological membranes causing perturbations in the bilayer and also by altering the physicochemical properties of membrane lipids. But, chronic alcohol consumption also increases nitric oxide (NO) production. There was no systemic study was done related to alcohol-induced production of NO and consequent formation of peroxynitrite mediated changes in biophysical and biochemical properties, structure, composition, integrity and function of erythrocyte membranes in chronic alcoholics. Hence, keeping all these conditions in mind the present study was undertaken to investigate the role of over produced nitric oxide on red cell membrane physicochemical properties in chronic alcoholics. Human male volunteers aged 44 ± 6 years with similar dietary habits were divided into two groups, namely nonalcoholic controls and chronic alcoholics (~125 g of alcohol at least five times per week for the past 10–12 years). Elevated nitrite and nitrate levels in plasma and lysate, changes in erythrocyte membrane individual phospholipid composition, increased lipid peroxidation, protein carbonyls, cholesterol and phospholipids ratio (C/P ratio) and anisotropic value (γ) with decreased sulfhydryl groups and Na⁺/K⁺-ATPase activity in alcoholics was evident from this study. RBC lysate NO was positively correlated with C/P ratio (r = 0.547) and anisotropic (γ) value (r = 0.428), Na⁺/K⁺-ATPase activity was negatively correlated with RBC lysate NO (r = ?0.372) and anisotropic (γ) value (r = ?0.624) in alcoholics. Alcohol-induced overproduction of nitric oxide reacts with superoxide radicals to produce peroxynitrite, which appears to be responsible for changes in erythrocyte membrane lipids and the activity of Na⁺/K⁺-ATPase.     

Seasonal variations in larval biomass and biochemical composition of brown shrimp, Crangon crangon (Decapoda, Caridea), at hatching

The “brown shrimp”, Crangon crangon (Linnaeus 1758), is a benthic key species in the North Sea ecosystem, supporting an intense commercial fishery. Its reproductive pattern is characterized by a continuous spawning season from mid-winter to early autumn. During this extended period, C. crangon shows significant seasonal variations in egg size and embryonic biomass, which may influence larval quality at hatching. In the present study, we quantified seasonal changes in dry weight (W) and chemical composition (CHN, protein and lipid) of newly hatched larvae of C. crangon. Our data revealed significant variations, with maximum biomass values at the beginning of the hatching season (February–March), a decrease throughout spring (April–May) and a minimum in summer (June–September). While all absolute values of biomass and biochemical constituents per larva showed highly significant differences between months (P < 0.001), CHN, protein and lipid concentrations (expressed as percentage values of dry weight) showed only marginally significant differences (P < 0.05). According to generalized additive models (GAM), key variables of embryonic development exerted significant effects on larval condition at hatching: The larval carbon content (C) was positively correlated with embryonic carbon content shortly after egg-laying (r ² = 0.60; P < 0.001) and negatively with the average incubation temperature during the period of embryonic development (r ² = 0.35; P < 0.001). Additionally, water temperature (r ² = 0.57; P < 0.001) and food availability (phytoplankton C; r ² = 0.39; P < 0.001) at the time of hatching were negatively correlated with larval C content at hatching. In conclusion, “winter larvae” hatching from larger “winter eggs” showed higher initial values of biomass compared to “summer larvae” originating from smaller “summer eggs”. This indicates carry-over effects persisting from the embryonic to the larval phase. Since “winter larvae” are more likely exposed to poor nutritional conditions, intraspecific variability in larval biomass at hatching is interpreted as part of an adaptive reproductive strategy compensating for strong seasonality in plankton production and transitory periods of larval food limitation.     

Advanced glycation end products (AGEs) are cross-sectionally associated with insulin secretion in healthy subjects

It has been postulated that chronic exposure to high levels of advanced glycation end products (AGEs), in particular from dietary sources, can impair insulin secretion. In the present study, we investigated the cross-sectional relationship between AGEs and acute insulin secretion during an intravenous glucose tolerance test (IVGTT) and following a 75 g oral glucose tolerance test (OGTT) in healthy humans. We report the cross-sectional association between circulating AGE concentrations and insulin secretory function in healthy humans (17 F: 27 M, aged 30 ± 10 years) with a wide range of BMI (24.6–31.0 kg/m²). Higher circulating concentrations of AGEs were related to increased first phase insulin secretion during IVGTT (r = 0.43; p < 0.05) and lower 2-h glucose concentrations during OGTT (r = ?0.31; p < 0.05). In addition, fasting (r = ?0.36; p < 0.05) and 2-h glucose concentrations were negatively related to circulating levels of soluble receptor for AGE (RAGE) isoforms (r = ?0.39; p < 0.01). In conclusion, in healthy humans, we show a cross-sectional association between advanced glycation end products and acute insulin secretion during glucose tolerance testing.     

Subcellular Localization and Kinetic Characterization of a Gill (Na+, K+)-ATPase from the Giant Freshwater Prawn Macrobrachium rosenbergii

The stimulation by Mg²⁺, Na⁺, K⁺, NH₄ ⁺, and ATP of (Na⁺, K⁺)-ATPase activity in a gill microsomal fraction from the freshwater prawn Macrobrachium rosenbergii was examined. Immunofluorescence labeling revealed that the (Na⁺, K⁺)-ATPase α-subunit is distributed predominantly within the intralamellar septum, while Western blotting revealed a single α-subunit isoform of about 108 kDa M _r. Under saturating Mg²⁺, Na⁺, and K⁺ concentrations, the enzyme hydrolyzed ATP, obeying cooperative kinetics with V _M = 115.0 ± 2.3 U mg^?1, K _0.5 = 0.10 ± 0.01 mmol L^?1. Stimulation by Na⁺ (V _M = 110.0 ± 3.3 U mg^?1, K _0.5 = 1.30 ± 0.03 mmol L^?1), Mg²⁺ (V _M = 115.0 ± 4.6 U mg^?1, K _0.5 = 0.96 ± 0.03 mmol L^?1), NH₄ ⁺ (V _M = 141.0 ± 5.6 U mg^?1, K _0.5 = 1.90 ± 0.04 mmol L^?1), and K⁺ (V _M = 120.0 ± 2.4 U mg^?1, K _M = 2.74 ± 0.08 mmol L^?1) followed single saturation curves and, except for K⁺, exhibited site–site interaction kinetics. Ouabain inhibited ATPase activity by around 73 % with K _I = 12.4 ± 1.3 mol L^?1. Complementary inhibition studies suggest the presence of F₀F₁–, Na⁺-, or K⁺-ATPases, but not V(H⁺)- or Ca²⁺-ATPases, in the gill microsomal preparation. K⁺ and NH₄ ⁺ synergistically stimulated enzyme activity (≈25 %), suggesting that these ions bind to different sites on the molecule. We propose a mechanism for the stimulation by both NH₄ ⁺, and K⁺ of the gill enzyme.     

Ontogeny of hypoxic modulation of cardiac performance and its allometry in the African clawed frog Xenopus laevis

The ontogeny of cardiac hypoxic responses, and how such responses may be modified by rearing environment, are poorly understood in amphibians. In this study, cardiac performance was investigated in Xenopus laevis from 2 to 25 days post-fertilization (dpf). Larvae were reared under either normoxia or moderate hypoxia (PO₂ = 110 mmHg), and each population was assessed in both normoxia and acute hypoxia. Heart rate (f _h ) of normoxic-reared larvae exhibited an early increase from 77 ± 1 beats min^?1 at 2 dpf to 153 ± 1 beats min^?1 at 4 dpf, followed by gradual decreases to 123 ± 3 beats min^?1 at 25 dpf. Stroke volume (SV), 6 ± 1 nl, and cardiac output (CO), 0.8 ± 0.1 μl min^?1, at 5 dpf both increased by more than 40-fold to 25 dpf with rapid larval growth (~30-fold increase in body mass). When exposed to acute hypoxia, normoxic-reared larvae increased f _h and CO between 5 and 25 dpf. Increased SV in acute hypoxia, produced by increased end-diastolic volume (EDV), only occurred before 10 dpf. Hypoxic-reared larvae showed decreased acute hypoxic responses of EDV, SV and CO at 7 and 10 dpf. Over the period of 2–25 dpf, cardiac scaling with mass showed scaling coefficients of ?0.04 (f _h ), 1.23 (SV) and 1.19 (CO), contrary to the cardiac scaling relationships described in birds and mammals. In addition, f _h scaling in hypoxic-reared larvae was altered to a shallower slope of ?0.01. Collectively, these results indicate that acute cardiac hypoxic responses develop before 5 dpf. Chronic hypoxia at a moderate level can not only modulate this cardiac reflex, but also changes cardiac scaling relationship with mass.     

Scaling of standard metabolic rate in estuarine crocodiles Crocodylus porosus

Standard metabolic rate (SMR, ml O₂ min^?1) of captive Crocodylus porosus at 30 °C scales with body mass (kg) according to the equation, SMR = 1.01 M^0.829, in animals ranging in body mass of 3.3 orders of magnitude (0.19–389 kg). The exponent is significantly higher than 0.75, so does not conform to quarter-power scaling theory, but rather is likely an emergent property with no single explanation. SMR at 1 kg body mass is similar to the literature for C. porosus and for alligators. The high exponent is not related to feeding, growth, or obesity of captive animals. The log-transformed data appear slightly curved, mainly because SMR is somewhat low in many of the largest animals (291–389 kg). A 3-parameter model is scarcely different from the linear one, but reveals a declining exponent between 0.862 and 0.798. A non-linear model on arithmetic axes overestimates SMR in 70 % of the smallest animals and does not satisfactorily represent the data.     

No Differences in Soil Carbon Stocks Across the Tree Line in the Peruvian Andes

Reliable soil organic carbon (SOC) stock measurements of all major ecosystems are essential for predicting the influence of global warming on global soil carbon pools, but hardly any detailed soil survey data are available for tropical montane cloud forests (TMCF) and adjacent high elevation grasslands above (puna). TMCF are among the most threatened of ecosystems under current predicted global warming scenarios. We conducted an intensive soil sampling campaign extending 40 km along the tree line in the Peruvian Andes between 2994 and 3860 m asl to quantify SOC stocks of TMCF, puna grassland, and shrubland sites in the transition zone between the two habitats. SOC stocks from the soil surface down to the bedrock averaged (±standard error SE) 11.8 (±1.5, N = 24) kg C/m² in TMCF, 14.7 (±1.4, N = 9) kg C/m² in the shrublands and 11.9 (±0.8, N = 35) kg C/m² in the grasslands and were not significantly different (P > 0.05 for all comparisons). However, soil profile analysis revealed distinct differences, with TMCF profiles showing a uniform SOC distribution with depth, shrublands a linear decrease, and puna sites an exponential decrease in SOC densities with soil depth. Organic soil layer thickness reached a maximum (~70 cm) at the upper limit of the TMCF and declined with increasing altitude toward puna sites. Within TMCF, no significant increase in SOC stocks with increasing altitude was observed, probably because of the large variations among SOC stocks at different sites, which in turn were correlated with spatial variation in soil depth.     

Evaluation of annual resolution coral geochemical records as climate proxies in the Great Barrier Reef of Australia

Sampling of annually banded massive coral skeletons at annual (or higher) resolutions is increasingly being used to obtain replicate long-term time series of changing seawater conditions. However, few of these studies have compared and calibrated the lower annual resolution records based on coral geochemical tracers with the corresponding instrumental climate records, although some studies have inferred the climatic significance of annual coral series derived from averages of monthly or sub-annual records. Here, we present annual resolution analysis of coral records of elemental and stable isotopic composition that are approximately 70 years long. These records were preserved in two coexisting colonies of Porites sp. from Arlington Reef, on the Great Barrier Reef in Australia, and are used to evaluate the climatic significance of annually resolved coral geochemical proxies. The geochemical records of coral sample “10AR2,” with its faster and relatively constant annual growth rate, appear to have been independent of skeletal growth rate and other vital effects. The annual resolution of Sr/Ca and Δδ¹⁸O time series was shown to be a good proxy for annual sea surface temperature (SST; r = ?0.67, n = 73, p < 0.0000001) and rainfall records (r = ?0.34, n = 67, p < 0.01). However, a slower growing coral sample, “10AR1” showed significantly lower correlations (r = ?0.20, n = 71, p = 0.05 for Sr/Ca and SST; r = ?0.19, n = 67, p = 0.06 for Δδ¹⁸O and rainfall), indicating its greater susceptibility to biological/metabolic effects. Our results suggest that while annually resolved coral records are potentially a valuable tool for determining, in particular, long timescale climate variability such as Pacific Decadal Oscillation, Interdecadal Pacific Oscillation, and other climatic factors, the selection of the coral sample is important, and replication is essential.     

Influence of the irradiance on phenols content and rooting of Ilex paraguariensis cuttings collected from adult plants

The influence of irradiance on phenolics contents and rooting of Ilex paraguariensis cuttings was studied. Results of the first experiment with stock plants under controlled-irradiance conditions show that when the irradiance level increased from 1.5 to 100 % PPFD, the oxidation of cuttings raised from 19 ± 11 to 88 ± 4 % (r ² = 0.64). At the same time, a strong correlation was observed between total phenolics content and irradiance (r ² = 0.7). In consequence, adventitious rooting diminished from 67 ± 5 to 3 ± 3 % under full radiation (r ² = 0.7). In the second experiment with stock plants subjected to field conditions, the results showed that the rooting process is strongly affected by the genotype (P < 0.0001), while the statistical analysis did not show a correlation between rooting and age of the donor plant. Season had a variable effect and depends on genotype. Although we did not find correlations between the rooting ability and the canopy structure of the stock plants, the position of the branches in the mother plant affected rooting and depended on season in addition to genotype. Concomitantly, the levels of soluble phenolics compounds were higher from leaves subjected to high-irradiance conditions than samples collected from inner canopy; which was coincident with the pattern of cuttings oxidation. In conclusion, our results provide evidences which support the hypothesis that the physiological status of the stock plant at the time that cuttings are excised is of utmost importance for the subsequent rooting of I. paraguariensis cuttings. The influence on soluble phenolics content of different irradiances given to the stock plants negatively affect the rooting process since the product of its oxidation cause the browning and death of the cuttings.     

Calibration of the Normal Cutoff Values of Systolic Dyssynchrony of the Left Ventricular Synchronicity in Normal Subjects Using Real-Time 3-Dimensional Echocardiography and the Effects of Age and Heart Rate

The normal data of left ventricular (LV) synchronicity by real-time 3-dimensional echocardiography (RT3DE) are lacking. We assessed the normal range/cutoff values of LV dyssynchrony parameters by RT3DE. For this purpose, RT3DE was performed in 130 healthy subjects, aged 53 ± 12 years. Time to the point of minimal regional systolic volume (Tmsv) was measured from time–volume curves in each segment. Standard deviation (SD) and maximal difference (Dif) of Tmsv were calculated from 16 (6 basal/6 mid/4 apical), 12 (6 basal/6 mid), and 6 (basal) LV segments together with the corresponding parameters adjusted by R–R interval. The data show non-significant difference between Tmsv-16-SD (9.24 ± 3.54 ms) and Tmsv-12-SD (8.80 ± 3.82 ms); with a correlation between two by both unadjusted (ms; r = 0.87) and adjusted (%R–R; r = 0.84) methods (P < 0.001). Heart rate correlated negatively with Tmsv (r = ?0.13 to ?0.34, P < 0.05–0.001) but had no effect on parameters adjusted for %R–R. Age and gender did not affect any of these parameters. Inter-observer variability was 3.3–4.6 % for 16, 4.8–9.1 % for 12, and 14.4–19.7 % for 6 segments. Thus, RT3DE is a reliable technique for detecting LV systolic dyssynchrony whereas the heart rate, but not age and gender, affects Tmsv parameters. Dyssynchrony parameters by 16 or 12 segments are superior to 6 segments in yielding comprehensive information and lower variability.     

Gross primary productivity of phytoplankton and planktonic respiration in inland floodplain wetlands of southeast Australia: habitat-dependent patterns and regulating processes

Gross primary productivity (GPP) of phytoplankton and planktonic respiration (PR) (i.e., planktonic metabolism) are critical pathways for carbon transformation in many aquatic ecosystems. In inland floodplain wetlands with variable inundation regimes, quantitative measurements of GPP and PR are rare and their relationships with wetland environmental conditions are largely unknown. We measured PR and the GPP of phytoplankton using light and dark biological oxygen demand bottles in open waters of channel and non-channel floodplain habitats of inland floodplain wetlands of southeast Australia that had been inundated by environmental water. Overall, GPP varied from 3.7 to 405.5 mg C m^?3 h^?1 (mean ± standard error: 89.4 ± 9.2 mg C m^?3 h^?1, n = 81), PR from 1.5 to 251.6 mg C m^?3 h^?1 (43.2 ± 5.6 mg C m^?3 h^?1, n = 81), and GPP/PR from 0.2 to 15.6 (3.0 ± 0.3, n = 81). In terms of wetland environmental conditions, total nitrogen (TN) ranged from 682.0 to 14,700.0 mg m^?3 (mean ± standard error: 2,643.0 ± 241.6 mg m^?3, n = 81), total phosphorus (TP) from 48.0 to 1,405.0 mg m^?3 (316.8 ± 31.4 mg m^?3, n = 81), and dissolved organic carbon (DOC) from 1.9 to 46.3 g m^?3 (22.0 ± 1.6 g m^?3, n = 81). Using ordinary least-squares multiple regression analyses, the rates of GPP and PR, and their ratio (GPP/PR) were modeled as a function of TN, TP, and DOC that had been measured concomitantly. The “best” models predicted GPP and GPP/PR ratio in channel habitats as a function of DOC; and GPP, PR, and GPP/PR in non-channel floodplain habitats as a function of TN and/or TP. The models explained between 46 and 74 % of the variance in channel habitats and between 17 and 87 % of the variance in non-channel floodplain habitats. Net autotrophy (mean GPP/PR 3.0) of planktonic metabolism in our work supports the prevailing view that wetlands are a net sink for carbon dioxide. We propose a nutrient-DOC framework, combined with hydrological and geomorphological delineations, to better predict and understand the planktonic metabolism in inland floodplain wetlands.     

Analysis of the reproductive cycle of female wild marble trout <Emphasis Type="Italic">Salmo marmoratus</Emphasis> in a prealpine brook of the Soča River basin (Northeast Italy)

The reproductive cycle of female wild marble trout Salmo marmoratus was studied in a prealpine watercourse (Northeast Italy). Gonadosomatic index, oocyte growth and plasma levels of 17β-oestradiol and testosterone were bimonthly measured during one year to obtain a detailed dataset and to gain useful tools for the identification of the reproductive stage without impact for the species. Monitored features showed significant variations during the study period: gonadosomatic index and oocyte size increased slightly but significantly during the first part of the cycle (from February to June), while steroid levels remained quite constant; for all parameters, major increases were highlighted between August and the spawning season (which occurred from the middle of November to the middle of December). Ranges and trends observed for gonadosomatic index (from 0.87 ± 0.41% to 10.91 ± 3.37%) and oocyte diameter (from 0.692 ± 0.031 mm to 4.624 ± 0.208 mm) were generally in agreement with literature regarding salmonids, while plasma levels of 17β-oestradiol (from 0.214 ± 0.015 ng ml^?1 to 78.090 ± 23.882 ng ml^?1) and testosterone (from 0.327 ± 0.086 ng ml^?1 to 71.800 ± 29.406 ng ml^?1) showed wider ranges. A strong non-linear relationship was found between oocyte size and 17β-oestradiol plasma concentration (r ² = 0.890) and especially between oocyte size and testosterone plasma concentration (r ² = 0.947). This last relationship could likely be used to obtain information relative to the gonadal development, especially in the case of an endangered species like Salmo marmoratus, which needs non-invasive tools for management.     

Reproduction efficiency of the Horse Leech,Haemopis sanguisuga (Linnaeus, 1758)

The aquaculture sector has recently focused on alternative food sources due to increased costs and limited resources. Live food sources, such as invertebrates, have important roles in digestion processes of fish. It was aimed to determine the reproduction efficiency of the horse leech, Haemopis sanguisuga, which is expected to be used as an alternative live nutritional source in aquaculture in this study. The study was conducted at ambient temperature in the laboratory conditions for three months. A total of 25 gravid leeches with an average body mass of 6.91 ± 2.20 g were used. The leeches lost up to 75% of their body mass and laid 4.30 ± 1.49 cocoons at 12.0 ± 8.2 days interval during cocoon laying period. Leeches stopped laying cocoon when the temperature fell to 16 °C. The average of offspring hatched from the cocoons was 6.45 ± 2.86. The length–body mass relationship of the offspring was exponential (W = 0.012L^2.809, R² = 0.898). The results from the current study indicate that the basic biological parameter affecting the reproduction efficiency of H. sanguisuga is the size of the broodstock leeches, and reproduction activity weakens the broodstocks.     

Size at maturity of Mediterranean marine fishes

In this review we collected data on the length at maturity (L_m) and maximum reported total length (L_max) of 565 Mediterranean marine fish stocks, representing 150 species, 68 families, 24 orders and 3 classes. Overall, L_m ranged from 2 cm, for the males of the toothcarp Aphanius fasciatus, to 350 cm, for the females of the bluntnose sixgill shark Hexanchus griseus. L_m was positively linearly related with L_max for Actinopterygii (logL_m = ?0.123 + 0.92 × logL_max; r ² = 0.87, n = 471, P < 0.001) and Elasmobranchii (logL_m = ?0.008 + 0.922 × logL_max; r ² = 0.90, n = 92, P < 0.001) with the two slopes being significantly different (ANCOVA: F = 2,904, P < 0.001). The reproductive load (L_m/L_max) ranged between 0.23 (sand steenbras Lithognathus mormyrus) and 0.94 (angular roughshark Oxynotus centrina and thornback ray Raja clavata). The mean L_m/L_max was significantly (ANOVA, F = 34.14, P < 0.001) lower for Actinopterygii (mean = 0.59, SD = 0.122, n = 471) compared to Elasmobranchii (mean = 0.70, SD = 0.132, n = 92) and Holocephali (mean = 0.77, SD = 0.077, n = 2). The L_m/L_max was significantly (ANOVA, F = 43.80, P < 0.001) higher for species providing some form of parental care, i.e. guarders, bearers, nesters (mean L_m/L_max ± SD = 0.68 ± 0.141, n = 111) compared to non-guarders (mean L_m/L_max ± SD = 0.59 ± 0.123, n = 454). The mean L_m/L_max displayed a remarkable constancy with longitude (northern and southern Mediterranean coastline: ANOVA, F = 0.01, P = 0.93), latitude (western, central and eastern regions: ANOVA, F = 1.25, P = 0.29) and habitat (ANOVA, F = 0.85, P = 0.51).     

The effectiveness of the high-LET radiations from the boron neutron capture [<Superscript>10</Superscript>B(n,α)<Superscript>7</Superscript>Li] reaction determined for induction of chromosome aberrations and apoptosis in lymphocytes of human blood samples

Provided that a selective accumulation of ¹⁰B-containing compounds is introduced in tumor cells, following irradiation by thermal neutrons produces high-LET alpha-particles (⁴He) and recoiling lithium-7 (⁷Li) nuclei emitted during the capture of thermalized neutrons (0.025 eV) from ¹⁰B. To estimate the biological effectiveness of this boron neutron capture [¹⁰B(n,α)⁷Li] reaction, the chromosome aberration assay and the flow cytometry apoptosis assay were applied. At the presence of the clinically used compounds BSH (sodium borocaptate) and BPA (p-boronophenylalanine), human lymphocytes were irradiated by sub-thermal neutrons. For analyzing chromosome aberrations, human lymphocytes were exposed to thermally equivalent neutron fluences of 1.82 × 10¹¹ cm^?2 or 7.30 × 10¹¹ cm^?2 (corresponding to thermal neutron doses of 0.062 and 0.248 Gy, respectively) in the presence of 0, 10, 20, and 30 ppm of BSH or BPA. Since the kerma coefficient of blood increased by 0.864 × 10^?12 Gy cm² per 10 ppm of ¹⁰B, the kerma coefficients in blood increase from 0.34 × 10^?12 cm² (blood without BSH or BPA) up to 2.93 × 10^?12 Gy cm² in the presence of 30 ppm of ¹⁰B. For the ¹⁰B(n, α)⁷Li reaction, linear dose–response relations for dicentrics with coefficients α = 0.0546 ± 0.0081 Gy^?1 for BSH and α = 0.0654 ± 0.0075 Gy^?1 for BPA were obtained at 0.062 Gy as well as α = 0.0985 ± 0.0284 Gy^?1 for BSH and α = 0.1293 ± 0.0419 Gy^?1 for BPA at 0.248 Gy. At both doses, the corresponding ¹⁰B(n, α)⁷Li reactions from BSH and BPA are not significantly different. A linear dose–response relation for dicentrics also was obtained for the induction of apoptosis by the ¹⁰B(n, α)⁷Li reaction at 0.248 Gy. The linear coefficients α = 0.0249 ± 0.0119 Gy^?1 for BSH and α = 0.0334 ± 0.0064 Gy^?1 for BPA are not significantly different. Independently of the applied thermal neutron doses of 0.062 Gy or 0.248 Gy, the ¹⁰B(n, α)⁷Li reaction from 30 ppm BSH or BPA induced an apparent RBE of about 2.2 for the production of dicentrics as compared to exposure to thermal neutrons alone. Since the apparent RBE value is defined as the product of the RBE of a thermal neutron dose alone times a boron localization factor which depends on the concentration of a ¹⁰B-containing compound, this localization factor determines the biological effectiveness of the ¹⁰B(n, α)⁷Li reaction.