Effect of gibberellic acid on the cyanobacterium <Emphasis Type="Italic">Nostoc linckia</Emphasis>

We investigated the influence of gibberellic acid (GA₃; 0, 1, 10, and 100 μM) on Nostoc linckia culture at 7, 14, and 21 days. The fresh and dry weight of N. linckia was increased considerably by the 10 and 100 μM GA₃ treatments. A reduction in heterocyst frequency was observed in cultures treated with 1 and 10 μM GA₃. Adding GA₃ to N. linckia culture had a little effect on cell size. The amount of chlorophyll a and carotenoids decreased at all concentrations of GA₃. The amount of phycocyanin increased up to twofold in 7-day-old culture treated with 1 μM GA₃, and similar changes were observed for allophycocyanin and phycoerythrin content after 7 days. The effect of GA₃ on reducing sugar content was different and was dependent on the growth period. A reduction in soluble sugar content was detected after GA₃ application in 7- and 14-day-old cyanobacteria. Cultures treated with GA₃ had a higher protein content after 14 days and a lower protein content after 7 and 21 days, and reduced nitrogenase activity after 7, 14, and 21 days. Our data show that GA₃ application can be a suitable and inexpensive way to increase N. linckia biomass and phycobiliprotein production.     

Rare case of osteomyelitis of tibial shaft caused by <Emphasis Type="Italic">Nocardia cyriacigeorgica</Emphasis>

Nocardiosis is a rare infection caused by the aerobic actinomycete of the Nocardia genus. In most cases, nocardiosis manifests as a lung infection or a bone lesion. Due to the nonspecific and mild clinical manifestations of nocardiosis, the establishment of definite diagnosis can be difficult. When antibiotic therapy is incorrectly targeted, only the symptoms of the disease are suppressed. The mainstay in the treatment of Nocardia osteomyelitis has so far been the combined surgical debridement with long-term, initially intravenous, antibiotic administration. We present the successful conservative treatment of a nocardiosis osteomyelitis of the tibia caused by the Nocardia cyriacigeorgica species in an 81-year-old female patient that manifested itself as a secondary affection on top of a primary nocardiosis infection of the lung. From microbiological examination, N. cyriacigeorgica was discovered; the identification was made using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) with an identification score of 1.9. The sensitivity was evaluated using E test. Sensitivity to trimethoprim/sulfamethoxazole, amikacin, imipenem, and linezolid was demonstrated. The bacteria were shown to be resistant to ciprofloxacin. For treatment, trimethoprim/sulfamethoxazole was used due to the value of minimum inhibitory concentration, which was 0.25 mg/L. The initial dose of 960 mg of trimethoprim/sulfamethoxazole every 8 h was reduced to 960 mg every 12 h after 3 months. The total duration of treatment was 7.5 months. Under the established treatment, the bone and lung lesions healed. Nocardiosis of the long bone is considered a rare disease and its precise diagnosis has not yet been standardized. We used the MALDI-TOF MS method for the identification of the causal organism which is a fast and reliable method according to current world literature even when compared with the rRNA genetic sequencing reference method. Our case study presents a rare case of osteomyelitis of tibial shaft caused by N. cyriacigeorgica and its successful conservative treatment.     

Influence of temperature on the effectiveness of a biogenic carbonate surface treatment for limestone conservation

So far, most studies on microbiologically induced carbonate precipitation for limestone conservation have been performed at temperatures optimal for the activity of the calcinogenic bacteria (i.e., 20–28 °C). Successful application in practice, however, requires adequate performance in a wide range of environmental conditions. Therefore, the aim of this study was to select microorganisms that are most suited for biodeposition at temperatures relevant for practice. In a first step, ureolytic microorganisms were screened for their growth and ureolytic activity at different temperatures (10, 20, 28, and 37 °C). Large differences in calcinogenic activity could be observed between experiments performed on agar plates and those performed in solution and in limestone. In a second step, the influence of temperature on the performance of the biodeposition treatment with different ureolytic microorganisms was evaluated, both on the consolidative and protective effect of the treatment. In contrast with the experiments on agar plates, the Sporosarcina psychrophila strains failed to produce significant amounts of calcium carbonate on limestone in conditions relevant for practice, even at 10 °C. This resulted in a poor performance of the treatment. From experiments performed on limestone prisms, it appeared that the mesophilic Bacillus sphaericus produced the highest amount of carbonate in the shortest amount of time at all temperatures tested. As a result, compared to the untreated specimens, the highest consolidative (64 % lower weight loss upon sonication) and protective effect (46 % decreased sorptivity) were observed for the treatments with this species. From this study, it appears that among all ureolytic strains tested, B. sphaericus is most suited for biodeposition applications in practice.     

Xylem anatomy and calculated hydraulic conductance of four <Emphasis Type="Italic">Nothofagus</Emphasis> species with contrasting distribution in South-Central Chile

Nothofagus obliqua, N. dombeyi, N. alpina and N. antarctica are characteristic tree species of the temperate forests on the western slopes of the Andes with centres of distribution that differ in their temperature and moisture regimes. We tested branch wood from co-occurring specimens of these species for the inherent differences in xylem anatomy and theoretical hydraulic conductance to evaluate their resistance to drought or frost. The hydraulic conductivity of the xylem was calculated using a modified Hagen–Poiseuille equation and related to wood density. Conduit dimensions were used to predict the water potential that would cause 50 % loss of hydraulic conductivity (Ψ ₅₀). Nothofagus alpina, which mainly grows at sites with low frost frequency, exhibited the largest conduits and the highest mean values for conduit area, fraction of conduit area in the cross-section and hydraulic conductivity, but the lowest wood density. Opposite relationships were found in the plastic N. antarctica, whose xylem seems to be least vulnerable to freezing-induced, but also to drought-induced embolism. Calculated Ψ ₅₀ was highest (least negative) in N. alpina, indicating a relatively high susceptibility to cavitation. The xylem of the thermophilic N. obliqua and of N. dombeyi, which mainly occurs under oceanic climate, but can also survive at sporadically dry and warm sites, is not particularly adapted to periods of drought stress. Across all species, wood density was negatively correlated with the calculated hydraulic conductance. The xylem traits of N. alpina might contribute to its relatively high growth rate and facilitate its spread into forest gaps.     

A multitrophic interaction: consequences of grazing on decomposition of needles of <Emphasis Type="Italic">Pinus pinea</Emphasis> L. by <Emphasis Type="Italic">Ommatoiulus sabulosus</Emphasis> L.

The aim of this study was to assess how the physical (needle fragmentation by trampling) and chemical (nutritional enrichment from faeces) changes brought about by grazing influence the consumption of needles of Pinus pinea L. by Ommatoiulus sabulosus L. Adult individuals of O. sabulosus were introduced into trays with pine needles subjected to four treatments: (i) intact needles, (ii) trampled needles, (iii) intact needles fertilized with liquid manure, and (iv) trampled needles fertilized with liquid manure. After 30 days, litter decomposition was determined as a function of mass loss over time. Biometric and chemical characteristics of the needles were also measured. The pine needle decomposition rate was significantly higher in the presence of O. sabulosus (20–40%) than in its absence, presenting in addition a significantly higher rate in the treatments enriched with nitrogen. Average needle length was significantly shorter in the trampled treatments (6.89 ± 0.50 cm) than in those with intact needles (11.95 ± 0.69 cm). With regard to leaf mass per area and resistance to breakage, no significant differences were found. The fertilized treatments presented significantly higher N content (50% higher) than those in the non-fertilized ones with a significantly lower C:N ratio. The results of the present study indicate that goat grazing (which produces both trampling and fertilization) favoured the consumption of pine needles by O. sabulosus, which thus influences, along with the microbial community, the decomposition rate of these needles and thereby promotes incorporation of N into the system.     

Shrub Expansion of <Emphasis Type="Italic">Alnus viridis</Emphasis> Drives Former Montane Grassland into Nitrogen Saturation

The N₂-fixing shrub Alnus viridis is currently encroaching on montane grasslands in the Alps as a result of reduced land management and complete abandonment. Alnus introduces large amounts of nitrogen (N) into these formerly N-poor grasslands and restricts the succession to montane forests. We studied pools and fluxes of N and the associated C pools in pastures (controls) and adjacent Alnus shrublands at two elevations (1650 versus 1950 m a.s.l.) in three valleys in the Swiss central Alps. The total N and C pools stored in 50-year-old Alnus shrubland did not exceed those in adjacent pastures with a total of approximately 610 g N m^?2 in phytomass plus soil (down to 30 cm) at both elevations. In Alnus stands, reduced soil N pools balanced the gain in phytomass N pools, a likely result of a faster turnover of soil N. The soil solution under Alnus was continuously enriched with nitrate, with a total N leaching of 0.79 g N m^?2 season^?1 (June–October) under 50-year-old stands at both elevations and the highest flux of 1.76 g N m^?2 season^?1 in 25-year-old shrubland at low elevation, clearly indicating an excess of available N in Alnus shrubland. In contrast, N leaching across all pastures was close to zero (0.08 g N m^?2) throughout the season. At the catchment scale, streamlet water showed increased nitrate concentrations with typical flushing peaks in spring and autumn, provided more than one fifth of the catchment area was covered by Alnus shrubs. We conclude that the expansion of Alnus rapidly converts centuries-old, N-poor grassland into N saturated shrubland, irrespective of elevation, and it reduces the C storage potential of the landscape because the Alnus dominance constrains re-establishment of a natural montane forest.     

Fate of ammonium 15N in a Norway spruce forest under long-term reduction in atmospheric N deposition

In the last decades, in particular forest ecosystems became increasingly N saturated due to elevated atmospheric N deposition, resulting from anthropogenic N emission. This led to serious consequences for the environment such as N leaching to the groundwater. Recent efforts to reduce N emissions raise the question if, and over what timescale, ecosystems recover to previous conditions. In order to study the effects on N distribution and N transformation processes under the lowered N deposition treatment, we investigated the fate of deposited NH₄ ⁺-¹⁵N in soil of a N-saturated Norway spruce forest (current N deposition: 34 kg ha^?1 year^?1; critical N load: 14 kg ha^?1 year^?1), where N deposition has been reduced to 11.5 kg ha^?1 year^?1 since 14.5 years. We traced the deposited ¹⁵N in needle litter, bulk soil, and amino acids, microbial biomass and inorganic N in soil. Under reduced N deposition, 123 ± 23% of the deposited N was retained in bulk soil, while this was only 72 ± 15% under ambient deposition. We presume that with reduced deposition the amount of deposited N was small enough to become completely immobilized in plant and soil and no leaching losses occurred. Trees receiving reduced N deposition showed a decline in N content as well as in ¹⁵N incorporation into needle litter, indicating reduced N plant uptake. In contrast, the distribution of ¹⁵N within the soil over active microbial biomass, microbial residues and inorganic N was not affected by the reduced N deposition. We conclude that the reduction in N deposition impacted only plant uptake and drainage losses, while microbial N transformation processes were not influenced. We assume changes in the biological N turnover to start with the onset of the decomposition of the new, N-depleted litter.     

Growth and nodulation in <Emphasis Type="Italic">Alnus sieboldiana</Emphasis> in response to <Emphasis Type="Italic">Frankia</Emphasis> inoculation and nitrogen treatments

Key message

We investigated a Frankia – Alnus sieboldiana symbiosis, including the minimum inoculum dose for constant nodulation, the period of time to nodulation after inoculation, and the effects of N on nodulation.

Abstract

Frankia is a nitrogen-fixing actinomycete that forms root nodules in some dicotyledonous plants, which are called actinorhizal. We studied nodule formation in Alnus sieboldiana, an actinorhizal plant, after inoculation with a Frankia isolate to establish techniques for Frankia inoculation and the cultivation of inoculated plants. Root nodules formed on seedlings of A. sieboldiana by 2 weeks after inoculation, and N₂ fixation measured by acetylene reduction activity started 3 weeks after inoculation. Nodulation was observed after inoculation with a Frankia isolate at 0.001 μL packed cell volume (pcv). The number of nodules formed on the seedlings inoculated with Frankia at more than 0.05 μL pcv was not significantly different. Nodule development and N₂ fixation were reduced when inoculated seedlings were treated weekly with 15 mM NH₄NO₃-N. In contrast, treatment with 3.75 or 0.9375 mM NH₄NO₃-N did not inhibit nodule development or N₂ fixation of inoculated seedlings by 15 weeks of N treatment.

     

Significant Improvement of Insulin Resistance of GK Rats by Treatment with Sodium Selenate

We studied the effect of sodium selenate on insulin resistance of Goto-Kakizaki (GK) rats. Rats were kept on standard laboratory chow with and without i.p. injections of sodium selenate (0.173 mg/kg body weight) for 14 days, and then subjected to the glucose clamp. The glucose clamp studies confirmed an improvement in insulin-stimulated glucose disposal in GK rats treated with sodium selenate, with respect to both insulin sensitivity and responsiveness. This amelioration of insulin resistance may be partly due to a direct effect of the sodium selenate on peripheral tissues. 2-Deoxyglucose uptake in sodium selenate-treated adipocytes was increased and the insulin findings suggest that sodium selenate increases not only insulin sensitivity but also insulin responsiveness. Sodium selenate also accelerated glucose incorporation into adipocytes of rats, suggesting that the action of sodium selenate is peripheral. Interestingly, sodium selenate at a high concentration (about 1 mmol/L) was more effective than insulin in enhancing glucose uptake. The present study suggested that sodium selenate treatment led to substantial improvement in peripheral insulin resistance.     

Forced hot-air treatment against <Emphasis Type="Italic">Bactrocera papayae</Emphasis> (Diptera: Tephritidae) in papaya

The Asian papaya fruit fly, Bactrocera papayae Drew and Hancock, was treated with hot-water immersion and forced hot air to develop a phytosanitary heat treatment schedule. Hot-water immersion tests were conducted with 12- and 24-h-old eggs and with first and third instar larvae to compare the relative thermotolerances of this fruit fly among these life stages. The 24-h-old eggs, the most thermotolerant among the four life stages tested, were subjected to time and temperature tests using cage-infested papaya fruits in a forced hot-air chamber. Heating the papayas to a minimum core temperature of 47.7 °C (95% confidence interval 47.2–48.3 °C) was estimated to induce probit-nine mortality based on a probit analysis of the data. Confirmatory tests in which papayas infested with 24-h-old eggs were heated to a minimum fruit core temperature of 47.2 °C that was maintained for 0–30 min followed by hydrocooling to a fruit core temperature of ≈25 °C resulted in the complete mortality of an estimated treated population of 43,425 eggs aged 24 h (99.9931% mortality at the 95% confidence level). Therefore, heating the papaya fruits to a core temperature of 47.2 °C for a minimum dwell time of 30 min, which was the longest dwell time in the confirmatory tests, may serve as a phytosanitary heat treatment schedule for the control of B. papayae in papaya fruits.     

Assessment of antibiotic resistance in <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> exposed to sequential in vitro antibiotic treatments

Background

Bacteria treated with different classes of antibiotics exhibit changes in susceptibility to successive antibiotic treatments. This study was designed to evaluate the influence of sequential antibiotic treatments on the development of antibiotic resistance in Klebsiella pneumoniae associated with β-lactamase and efflux pump activities.

Methods

The antibiotic susceptibility, β-lactamase activity, and efflux activity were determined in K. pneumoniae grown at 37 °C by adding initial (0 h) and second antibiotics (8 or 12 h). Treatments include control (CON; no first and second antibiotic addition), no initial antibiotic addition followed by 1 MIC ciprofloxacin addition (CON-CIP), no initial antibiotic addition followed by 1 MIC meropenem addition (CON-MER), initial 1/4 MIC ciprofloxacin addition followed by no antibiotic addition (1/4CIP-CON), initial 1/4 MIC ciprofloxacin addition followed by 1 MIC ciprofloxacin addition (1/4CIP-CIP), and initial 1/4 MIC ciprofloxacin addition followed by 1 MIC meropenem addition (1/4CIP-MER).

Results

Compared to the CON, the initial addition of 1/4 MIC ciprofloxacin inhibited the growth of K. pneumoniae throughout the incubation period. The ciprofloxacin treatments (CON-CIP and 1/4CIP-CIP) showed significant reduction in the number of K. pneumoniae cells compared to meropenem (CON-MER and 1/4CIP-MER). The 1/4CIP-CIP achieved a further 1 log reduction of K. pneumoniae, when compared to the 1/4CIP-CON and 1/CIP-MER. The increase in sensitivity of K. pneumoniae to cefotaxime, kanamycin, levofloxacin, nalidixic acid was observed for CON-CIP. Noticeable cross-resistance pattern was observed at the 1/4CIP-CIP, showing the increased resistance of K. pneumoniae to chloramphenicol, ciprofloxacin, kanamycin, levofloxacin, nalidixic acid norfloxacin, sulphamethoxazole/trimethoprim, and tetracycline. The levels of β-lactamase activities were estimated to be 8.4 μmol/min/ml for CON, 7.7 μmol/min/ml for 1/4CIP-CON and as low as 2.9 μmol/min/ml for CON-CIP. Compared to the absence of phenylalanine-arginine-β-naphthylamide (PAβN), the fluorescence intensity of EtBr was increased in K. pneumoniae cells treated at the CON, CON-CIP, and CON-MER in the presence of PAβN. However, the efflux pump activity remained in K. pneumoniae cells treated at the 1/CIP, 1/CIP–CIP, and 1/CIP-MER in the presence of PAβN.

Conclusion

The results suggest that the pre-exposed antibiotic history, treatment order, and concentrations influenced the development of multiple antibiotic resistant associated with β-lactamase and efflux pump activities. This study highlights the importance of antibiotic treatment conditions, which would be taken into consideration when new antibiotic strategy is designed to prevent antibiotic resistance.

     

Testing latitudinal patterns of tidepool fish assemblages: local substrate characteristics affect regional-scale trends

We tested the regional-scale latitudinal patterns of occurrence and community structure of rocky tidepool fishes in relation to local environmental factors, particularly substrate characteristics. Data were derived from intensive field observations conducted on 36 shore sites spread across ca. 1,000 km north–south (24°03′N–32°45′N) in the south-western Japan. While numbers of families, genera, species and individuals per unit area decreased with latitude, these were dependent on substrate types: sites with non-limestone rock substrates tended to harbour larger numbers than limestone sites at the same latitude. Relative abundances of two dominant families (Blenniidae and Gobiidae) varied among sites with weak latitudinal gradients. Species-specific trends of latitudinal distribution were observed in most of the common intertidal fishes, with over half of the species demonstrating substrate-dependent variation. Species composition was clearly different between the Kyushu Island and the Ryukyus and also between limestone and non-limestone sites. Thus, our results clearly demonstrated that the regional-scale latitudinal trends of tidepool fish assemblages were partially dependent on local environmental characteristics (substrate types). Consideration was given to the influences of the Kuroshio Current and other factors including species interactions that might have helped modify observed latitudinal patterns.     

The regulatory role of riboflavin in the drought tolerance of tobacco plants depends on ROS production

Riboflavin (vitamin B₂) is required for normal plant growth and development. Previous studies have shown that riboflavin application can enhance pathogen resistance in plants. Here, we investigated the role of riboflavin in increasing drought tolerance (10 % PEG6000 treatment) in plants. We treated 4 week-old tobacco plants with five different levels of riboflavin (0, 4, 20, 100 and 500 μM) for 5 days and examined their antioxidant responses and levels of drought tolerance. Compared with the controls, low and moderate levels of riboflavin treatment enhanced drought tolerance in the tobacco plants, whereas higher concentrations of riboflavin (500 μM) impaired drought tolerance. Further analysis revealed that plants treated with 500 μM riboflavin accumulated higher levels of ROS (O₂ ^? and H₂O₂) and lipid peroxide than the control plants or plants treated with low levels of riboflavin. Consistent with this observation, the activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) were higher in plants treated with low or moderate (4, 20 and 100 μM) levels of riboflavin compared with the control. We also found that chlorophyll degraded rapidly in control and 500 μM riboflavin-treated plants under drought stress conditions. In addition, the survival times of the riboflavin-treated plants were significantly modified by treatment with reduced glutathione, a well-known ROS scavenger, under drought stress conditions. Thus, riboflavin-mediated ROS production may determine the effects of riboflavin on drought tolerance in tobacco plants.     

Comparative transcriptome profiling of freezing stress responses in loquat (<Emphasis Type="Italic">Eriobotrya japonica</Emphasis>) fruitlets

Loquat (Eriobotrya japonica Lindl.) is an important subtropical, commercial fruit in China. It blossoms during autumn and winter in most areas of China and its fruitlets usually suffer from freezing stress. However, studies about the mechanisms underlying freezing stress in loquat are very limited. The gene expression profiles of loquat fruitlets subjected to freezing (G2 library) versus non-treated ones (G1 library) were investigated using Illumina sequencing technology to elucidate the molecular mechanisms and identify the genes that play vital roles in the freezing stress response. The results showed that approximately 157.63 million reads in total were obtained from freeze-treated and non-treated loquat fruitlets. These reads were assembled into 87,379 unigenes with an average length of 710 bp and an N50 of 1,200 bp. After comparing the profiles obtained from the G1 and G2 libraries, 2,892 differentially expressed genes were identified, of which 1,883 were up-regulated and 1,009 were down-regulated in the treated samples compared to non-treated ones. These unigenes showed significant differences in expression for carbohydrate transport and metabolism, amino acid metabolism, energy metabolism, and lipid metabolism, which are involved in defense against freezing stress. Glycolysis/gluconeogenesis was one of the most significantly regulated pathways. Freezing also significantly damaged the membrane system of loquat fruitlets, and several defense mechanisms were induced. Some selected genes related to low temperature resistance were validated by quantitative real-time PCR (qRT-PCR). The results revealed many genes and pathways that are part of freezing resistance processes and expand our understanding of the complex molecular events involved in freezing stress.     

Root decomposition of <Emphasis Type="Italic">Artemisia halodendron</Emphasis> and its effect on soil nitrogen and soil organic carbon in the Horqin Sandy Land,northeastern China

Root decomposition is a critical feedback from the plant to the soil, especially in sandy land where strong winds remove aboveground litter. As a pioneer shrub in semi-mobile dunes of the Horqin sandy land, Artemisia halodendron has multiple effects on nutrient capture and the microenvironment. However, its root decomposition has not been studied in terms of its influence on soil organic carbon (SOC) and nitrogen (N). In this study, we buried fine (≤2 mm) and coarse roots in litterbags at a depth of 15 cm below semi-mobile dunes. We measured the masses remaining and the C and N contents at intervals during 434 days of decomposition. The soils below the litterbags were then divided into layers and sampled to measure the SOC and N contents. After rapid initial decomposition, both coarse and fine roots decomposed slowly. After 53 days, 36.2 % of coarse roots and 39.8 % of fine roots had decomposed. In contrast, only 18.4 % of coarse roots and 30.5 % of fine roots decomposed in the following 381 days. Fine roots decomposed significantly faster, and their decomposition rate after the initial rapid decay was strongly related to climate (R ² = 0.716, P < 0.05). Root decomposition increased SOC and N contents below the litterbags, with larger effects for fine roots. The SOC content was more variable between soil layers than the N content. Thus, decomposition of A. halodendron roots cannot be ignored when studying SOC and N feedbacks from plants to the soil, particularly for fine roots.     

Blood clearance and occupational exposure for <Superscript>177</Superscript>Lu-DOTATATE compared to <Superscript>177</Superscript>Lu-PSMA radionuclide therapy

The main target of this work is to examine blood clearance and external exposure for ¹⁷⁷Lu-DOTATATE compared with new emerging ¹⁷⁷Lu-PSMA therapy. Blood clearance and radiation exposure of 31 patients treated with 5.5?±?1.1 GBq ¹⁷⁷Lu-DOTATATE were compared to those of 23 patients treated with 7.4 GBq ¹⁷⁷Lu-PSMA. Dose rates were measured at several distances and time points up to 120 h after treatment. Blood samples were collected conjunctively after infusion. Caregiver’s cumulative dose was measured by means of an OSL (optically stimulated luminescence) dosimeter for 4–5 days and medical staff’s dose was also estimated using electronic personal dosimeters. Finger dose was determined via ring TLD (Thermoluminescence Dosimeter) for radiopharmacists and nurses. Dose rates due to ¹⁷⁷Lu-DOTATATE at a distance of 1 m, 4 h and 6 h after infusion, were 3.0?±?2.8 and 2?±?1.9 µSv/(h GBq), respectively, while those due to ¹⁷⁷Lu-PSMA were 3.1?±?0.8 and 2.2?±?0.9 µSv/(h GBq). Total effective dose of 17 caregivers was 100–200 µSv for ¹⁷⁷Lu-DOTATATE therapy. Mean effective doses to nurses and radiopharmacists were 5 and 4 µSv per patient, respectively, while those for physicists and physicians were 2 µSv per patient. For ¹⁷⁷Lu-DOTATATE, effective half-life in blood and early elimination phase were 0.31?±?0.13 and 4.5?±?1 h, while they were found as 0.4?±?0.1 and 5?±?1 h, respectively, for ¹⁷⁷Lu-PSMA. The first micturition time following ¹⁷⁷Lu-DOTATATE infusion was noted after 36?±?14 min, while the second and third voiding times were after 74?±?9 and 128?±?41 min, respectively. It is concluded that blood clearance and radiation exposure for ¹⁷⁷Lu-DOTATATE are very similar to those for ¹⁷⁷Lu-PSMA, and both treatment modalities are reasonably reliable for outpatient treatment, since the mean dose rate [2.1 µSv/(h GBq)] decreased below the dose rate that allows release of the patient from the hospital (20 µSv/h) after 6 h at 1 m distance.     

Betaine Down Regulates Apelin Gene Expression in Cardiac and Adipose Tissues of Insulin Resistant Diabetic Rats Fed by High-Calorie Diet

Apelin is a newly discovered peptide that its serum level increases in diabetic patients with cardiovascular dysfunction. Recent studies indicate the beneficial actions of betaine in reducing the cardiovascular and metabolic complications, however data related to its effect on adipocytokine expression is limited. The aim of this study was to evaluate the effect of betaine supplementation on Apelin gene expression in cardiac muscle and adipose tissue of insulin resistance, diabetic rats fed by a high calorie diet. To induce insulin resistance rats were fed with high fat/high carbohydrate diet for five weeks and then 30 mg/kg STZ was injected intraperitoneally. After confirming of diabetes incidence (serum glucose above 7.5 mmol/l) the animals were treated with 1 % betaine in drinking water for 28 days. At days 14 and 28 after treatment, animals were euthanized and Apelin gene expression was evaluated by real time PCR and western blot in heart and adipose tissues. Serum levels of insulin, Apelin and glucose and HOMA–IR were also measured. Our results showed that feeding of rats by a high calorie diets caused insulin resistance, which was manifested by elevated plasma insulin, glucose and Apelin levels and also HOMA–IR. Apelin gene expression in heart and adipose tissues were significantly increased simultaneously with the progression of diabetes. Betaine supplementation decreased serum Apelin and down regulated Apelin expression in adipose tissue and cardiac muscle, particularly at day 28 of treatment. We concluded that betaine might improve metabolic and cardiovascular complications in diabetic patients by regulation of Apelin expression and secretion.     

An <Emphasis Type="Italic">in vitro</Emphasis> model for the development of acquired tamoxifen resistance

The development of resistance to tamoxifen (Tam) remains a challenging clinical problem for ER+ breast cancer patients. To understand the mechanisms underlying of resistance, previous studies have driven the acquisition of Tam resistance by exposing cells to varying concentration of drug for varying lengths of time. However, a detailed protocol for the establishment of Tam-resistant cells remains to be clarified. In the present study, we aimed to determine and compare the effect of different in vitro protocols on the degree of resistance to 4-hydroxytamoxifen (4-OH Tam) for MCF7 cells. For this purpose, MCF7-Tam resistance (MCF7-TamR) cells were developed by treated with different concentrations (100, 200, 400, 600, 800 and 1000 nM) of 4-OH Tam over 3 months. The relative resistance was measured by WST-1 analysis. Studies characterizing of the 4-OH Tam resistance of MCF7-TamR cells were performed by 17 β-oestradiol (E2) and Annexin V/PI analysis. In addition, the expression levels of ABCC1, ABCG2 and ABCG1 were detected by RT-PCR, any changes in morphological of each resistance group were observed at the end of each month and compared with parental MCF7 cells. Consequently, exposure time and concentration can affect the degree of resistance to 4-OH Tam; thus, dose and treatment duration should be chosen according to the desired degree of resistance. This work presents a novel procedure for the generation of MCF7-TamR cells, thus enabling the identification and characterization of MCF7-TamR cells.     

Contamination effects on soil density fractions from high N or C content sodium polytungstate

Sodium polytungstate (SPT) is currently the material of choice for soil density fractionation (DF). We recently detected high levels of N in several types of commercially available SPT (0.74–1.4 mg g^?1), raising a concern that undesirable chemical effects on soils may occur during the DF procedure. To address this concern, we conducted two experiments to examine effects of SPT on C and N in the resulting soil fractions. First we suspended A-horizon material from three soil types of greatly differing mineralogy for 24 h in solutions containing three types of commercially obtained SPT and commercial SPT that had been passed through cation exchange resin columns. We compared %C, %N, ¹⁵N and ¹³C values in treated and untreated soils. We also spiked SPT with tracer-level ¹⁵NH₄ ⁺ to measure potential NH₄ ⁺ absorption by the soil fractions. Results suggest that the N-rich commercially available SPT can have a considerable effect on δ¹⁵N values, likely due to the presence of ¹⁵N-enriched NH₄ ⁺ in the SPT. In one of our soils, ¹⁵N enrichment of 3‰ was observed associated with overnight soaking in N-rich SPT (0.74 mg g^?1). By contrast, when using SPT with low levels of N (0.05 mg g^?1), no significant changes in ¹⁵N were observed. The remaining soil (after suspension and rinsing) was similar in %C, %N, ¹⁵N and ¹³C to the untreated bulk soil, suggesting that suspension of soil in SPT with low N levels purchased from the manufacturer or else through treatment with cation exchange resins does not greatly alter these variables. Low-N SPT is available commercially although it must be specifically requested from the manufacturer and is currently more expensive to purchase. Our results confirm that SPT tested and known to be low in C and N (<0.06 mg g^?1) does not adversely contaminate soils during the soil density fractionation procedure. If using newly purchased or recycled SPT with higher N or C levels than this, we recommend thorough testing for possible contamination effects prior to use. However we caution against using SPT that contains N or C levels >0.5 mg g^?1.