The SnRK2.10 kinase mitigates the adverse effects of salinity by protecting photosynthetic machinery.

SNF1-Related protein kinases Type 2 (SnRK2) are plant-specific enzymes widely distributed across the plant kingdom. They are key players controlling abscisic acid (ABA)-dependent and ABA-independent signaling pathways in the plant response to osmotic stress. Here we established that SnRK2.4 and SnRK2.10, ABA-nonactivated kinases, are activated in Arabidopsis thaliana rosettes during the early response to salt stress and contribute to leaf growth retardation under prolonged salinity but act by maintaining different salt-triggered mechanisms. Under salinity, snrk2.10 insertion mutants were impaired in the reconstruction and rearrangement of damaged core and antenna protein complexes in photosystem II (PSII), which led to stronger non-photochemical quenching, lower maximal quantum yield of PSII, and lower adaptation of the photosynthetic apparatus to high light intensity. The observed effects were likely caused by disturbed accumulation and phosphorylation status of the main PSII core and antenna proteins. Finally, we found a higher accumulation of reactive oxygen species (ROS) in the snrk2.10 mutant leaves under a few-day-long exposure to salinity which also could contribute to the stronger damage of the photosynthetic apparatus and cause other deleterious effects affecting plant growth. We found that the snrk2.4 mutant plants did not display substantial changes in photosynthesis. Overall, our results indicate that SnRK2.10 is activated in leaves shortly after plant exposure to salinity and contributes to salt stress tolerance by maintaining efficient photosynthesis and preventing oxidative damage.

Radiotherapy-Induced Fatigue in Breast Cancer Patients.

BACKGROUND: A large proportion of breast cancer patients who undergo adjuvant radiotherapy suffer from radiotherapy-induced fatigue. The possible causative factors of this specific side effect are diverse. SUMMARY: Prevalence, duration, and severity of radiotherapy-induced fatigue are dependent on the type of radiotherapy, as well as on the irradiated volume, dose scheme, on the number of radiation fields, the combination with other treatments, diurnal rhythm, smoking, and time-to-hospitalization. Recommended treatments include non-pharmacologic interventions, such as physical and psychosocial interventions. Pharmacologic therapies include treatment with methylphenidate and modafinil. In addition to its early detection with standardized instruments, adequate education to breast cancer patients about risks and predisposing factors of radiotherapy-induced fatigue is essential. Multidimensional strategies help to maintain the patients' quality of life and therefore guarantee treatment adherence and efficacy. KEY MESSAGES: Radiotherapy-induced fatigue is an underreported, underdiagnosed, and undertreated side effect. This review provides an overview of radiotherapy-induced fatigue in breast cancer patients receiving adjuvant radiotherapy.

Expression of maize calcium-dependent protein kinase (ZmCPK11) improves salt tolerance in transgenic Arabidopsis plants by regulating sodium and potassium homeostasis and stabilizing photosystem II.

In plants, CALCIUM-DEPENDENT PROTEIN KINASES (CDPKs/CPKs) are involved in calcium signaling in response to endogenous and environmental stimuli. Here, we report that ZmCPK11, one of maize CDPKs, participates in salt stress response and tolerance. Salt stress induced expression and upregulated the activity of ZmCPK11 in maize roots and leaves. Activation of ZmCPK11 upon salt stress was also observed in roots and leaves of transgenic Arabidopsis plants expressing ZmCPK11. The transgenic plants showed a long-root phenotype under control conditions and a short-root phenotype under NaCl, abscisic acid (ABA) or jasmonic acid (JA) treatment. Analysis of ABA and JA content in roots indicated that ZmCPK11 can mediate root growth by regulating the levels of these phytohormones. Moreover, 4-week-old transgenic plants were more tolerant to salinity than the wild-type plants. Their leaves were less chlorotic and showed weaker symptoms of senescence accompanied by higher chlorophyll content and higher quantum efficiency of photosystem II. The expression of Na+ /K+ transporters (HKT1, SOS1 and NHX1) and transcription factors (CBF1, CBF2, CBF3, ZAT6 and ZAT10) with known links to salinity tolerance was upregulated in roots of the transgenic plants upon salt stress. Furthermore, the transgenic plants accumulated less Na+ in roots and leaves under salinity, and showed a higher K+ /Na+ ratio in leaves. These results show that the improved salt tolerance in ZmCPK11-transgenic plants could be due to an upregulation of genes involved in the maintenance of intracellular Na+ and K+ homeostasis and a protection of photosystem II against damage.

SNF1-Related Protein Kinases SnRK2.4 and SnRK2.10 Modulate ROS Homeostasis in Plant Response to Salt Stress.

In response to salinity and various other environmental stresses, plants accumulate reactive oxygen species (ROS). The ROS produced at very early stages of the stress response act as signaling molecules activating defense mechanisms, whereas those produced at later stages in an uncontrolled way are detrimental to plant cells by damaging lipids, DNA, and proteins. Multiple systems are involved in ROS generation and also in ROS scavenging. Their level and activity are tightly controlled to ensure ROS homeostasis and protect the plant against the negative effects of the environment. The signaling pathways responsible for maintaining ROS homeostasis in abiotic stress conditions remain largely unknown. Here, we show that in Arabidopsis thaliana, two abscisic acid- (ABA)-non-activated SNF1-releted protein kinases 2 (SnRK2) kinases, SnRK2.4 and SnRK2.10, are involved in the regulation of ROS homeostasis in response to salinity. They regulate the expression of several genes responsible for ROS generation at early stages of the stress response as well as those responsible for their removal. Moreover, the SnRK2.4 regulate catalase levels and its activity and the level of ascorbate in seedlings exposed to salt stress.

Adverse Events of Trastuzumab Emtansine (T-DM1) in the Treatment of HER2-Positive Breast Cancer Patients.

The human epidermal growth factor receptor 2 (HER2) is commonly associated with poor prognosis and is overexpressed in approximately 15-20% of all breast cancers. The introduction of HER2-targeted therapies led to significant improvement in the prognosis of patients with HER2-positive breast cancer, for both early and advanced disease. These targeted therapies include the antibodies trastzumab and pertuzumab, the tyrosine kinase inhibitor lapatinib, and the antibody-drug conjugate trastuzumab emtansine (T-DM1). T-DM1 combines the anti-tumor activity of trastuzumab with that of DM1, a highly potent derivative of the anti-microtubule agent maytansine, resulting in increased anti-tumor activity. Notably, this agent has been demonstrated to be safe and is associated with low toxicity rates. However, maytansinoid, the cytotoxic component of T-DM1, does have the potential to induce various adverse events, particularly radiation necrosis, when used in combination with stereotactic radiosurgery. In this review, we aimed to summarize the current literature regarding T-DM1 safety and toxicity, with special emphasis on the existing landmark studies.

Enzymatically Modified Starch Ameliorates Postprandial Serum Triglycerides and Lipid Metabolome in Growing Pigs.

Developing host digestion-resistant starches to promote human health is of great research interest. Chemically modified starches (CMS) are widely used in processed foods and although the modification of the starch molecule allows specific reduction in digestibility, the metabolic effects of CMS have been less well described. This short-term study evaluated the impact of enzymatically modified starch (EMS) on fasting and postprandial profiles of blood glucose, insulin and lipids, and serum metabolome in growing pigs. Eight jugular-vein catheterized pigs (initial body weight, 37.4 kg; 4 months of age) were fed 2 diets containing 72% purified starch (EMS or waxy corn starch (control)) in a cross-over design for 7 days. On day 8, an 8-hour meal tolerance test (MTT) was performed with serial blood samplings. Besides biochemical analysis, serum was analysed for 201 metabolites through targeted mass spectrometry-based metabolomic approaches. Pigs fed the EMS diet showed increased (P<0.05) immediate serum insulin and plasma glucose response compared to pigs fed the control diet; however, area-under-the-curves for insulin and glucose were not different among diets. Results from MTT indicated reduced postprandial serum triglycerides with EMS versus control diet (P<0.05). Likewise, serum metabolome profiling identified characteristic changes in glycerophospholipid, lysophospholipids, sphingomyelins and amino acid metabolome profiles with EMS diet compared to control diet. Results showed rapid adaptations of blood metabolites to dietary starch shifts within 7 days. In conclusion, EMS ingestion showed potential to attenuate postprandial raise in serum lipids and suggested constant alteration in the synthesis or breakdown of sphingolipids and phospholipids which might be a health benefit of EMS consumption. Because serum insulin was not lowered, more research is warranted to reveal possible underlying mechanisms behind the observed changes in the profile of serum lipid metabolome in response to EMS consumption.

Comparison of respiratory function during TIVA (romifidine, ketamine, midazolam) and isoflurane anaesthesia in spontaneously breathing ponies Part I: blood gas analysis and cardiorespiratory variables.

OBJECTIVE: To compare pulmonary function and gas exchange in ponies during maintenance of anaesthesia with isoflurane or by a total intravenous anaesthesia (TIVA) technique. STUDY DESIGN: Experimental, cross-over study. ANIMALS: Six healthy ponies weighing mean 286 (range 233-388) ± SD 61 kg, age 13 (9-16) ± 3 years. METHODS: The ponies were anaesthetized twice, a minimum of two weeks apart. Following sedation with romifidine [80 µg kg(-1) intravenously (IV)], anaesthesia was induced IV with midazolam (0.06 mg kg(-1)) and ketamine (2.5 mg kg(-1), then maintained either with inhaled isoflurane (Fe'Iso = 1.1 vol%) (T-ISO) or an IV infusion of romifidine (120 µg kg(-1) hour(-1)), midazolam (0.09 mg kg(-1) hour(-1) IV) and ketamine (3.3 mg kg(-1) hour(-1)) (T-TIVA). Ponies were placed in lateral recumbency. Breathing was spontaneous and Fi'O(2) 60%. After an instrumentation/stabilisation period of 30 minutes, arterial and mixed venous blood samples were taken simultaneously every 10 minutes for 60 minutes and analysed immediately. Oxygen extraction ratio (O(2)ER) and venous admixture were calculated. Tidal volume (TV), minute volume (MV), respiratory rate (f(R)), packed cell volume (PCV), arterial blood pressure and heart rate (HR) were measured and recorded. Data were analysed with mixed model anova (α = 0.05). Treatments were compared overall and at two selected time points (T30 and T60) using Bonferroni correction. RESULTS: Arterial and mixed venous partial pressures of O(2) and CO(2), and TV were significantly lower and MV and f(R) were higher in T-TIVA compared to T-ISO. Venous admixture did not differ between treatments. O(2) R was significantly higher in T-TIVA. Mean arterial pressure was higher and HR was lower in T-TIVA compared to T-ISO. CONCLUSIONS AND CLINICAL RELEVANCE: Whilst arterial CO(2) was within an acceptable range during both protocols, the impairment of oxygenation was more pronounced with the T-TIVA evidenced by lower arterial and venous oxygen partial pressures.

Is DNA methylation modulated by wounding-induced oxidative burst in maize?

Plants respond to environmental changes by modifying gene expression. One of the mechanisms regulating gene expression is methylation of cytosine to 5-methylcytosine (m(5)C) which modulates gene expression by changing chromatin structure. Methylation/demethylation processes affect genes that are controlled upon environmental stresses. Here, on account of the regulatory role of m(5)C, we evaluate the content of m(5)C in DNA from normal and wound-damaged maize leaves. Wounding leads to a transient decrease of the global DNA methylation level ca 20-30% 1 h after the treatment followed by a return to the initial level within the next hours. Similar results were obtained using of radio-labeled nucleotides separated by Thin Layer Chromatography (TLC) or using m(5)C-specific Enzyme-Linked Immunosorbent Assay (ELISA). Wounding induced in maize leaves a two-step oxidative stress, an early one just after wounding and the second two hours later. It coincides with the transient changes of the cytosine methylation level. In the stress-inducible maize calcium-dependent protein kinase ZmCPK11 gene wounding transiently reduced methylation of cytosines 100 and 126 in the first exon.

Comparison of respiratory function during TIVA and isoflurane anaesthesia in ponies Part II: breathing patterns and transdiaphragmatic pressure.

OBJECTIVE: To compare breathing patterns and transdiaphragmatic pressure during total intravenous (TIVA) and isoflurane anaesthesia in ponies. STUDY DESIGN: Experimental, cross-over study. ANIMALS: Six healthy ponies weighing 286 (233-388) ± 61 kg, age 13 (9-16) ± 3 years. METHODS: Following premedication with romifidine [80 µg kg(-1) intravenously (IV)], general anaesthesia was induced with midazolam (0.06 mg kg(-1) IV) and ketamine (2.5 mg kg(-1) IV) and maintained with either isoflurane (Fe'Iso = 1.1%) (T-ISO) or an IV combination of romifidine (120 µg kg(-1) per hour), midazolam (0.09 mg kg(-1) hour(-1)) and ketamine (3.3 mg kg(-1) hour(-1)) (T-TIVA), while breathing 60% oxygen (FIO(2)). The circumference changes of the rib cage (RC) and abdominal compartment (ABD) were recorded using respiratory ultrasonic plethysmography (RUP). Balloon tipped catheters were placed in the distal oesophagus and the stomach and maximal transdiaphragmatic pressure (Pdi max) was calculated during Mueller's manoeuvre. RESULTS: The breathing pattern T-ISO was more regular and respiratory rate significantly lower compared with T-TIVA. Ponies in T-TIVA showed regularly appearing sighs, which were never observed in T-ISO. Different contribution of the RC and ABD compartments to the breathing pattern was observed with a smaller participation of the RC to the total volume change during T-ISO. Transdiaphragmatic pressures (mean 13.7 ± SD 8.61 versus 23.4 ± 7.27 cmH(2) O, p < 0.0001) were lower in T-TIVA compared to T-ISO [corrected]. The sum of the RC and ABD circumferential changes was lower during T-TIVA compared to T-ISO (6.32 ± 4.42 versus 11.72 ± 4.38 units, p < 0.0001). CONCLUSION AND CLINICAL RELEVANCE: Marked differences in breathing pattern and transdiaphragmatic pressure exist during inhalation- and TIVA and these should be taken into account for clinical estimation of anaesthetic depth.

Variety of non-invasive continuous monitoring methodologies including electrical impedance tomography provides novel insights into the physiology of lung collapse and recruitment ­ case report of an anaesthetized horse.

INTRODUCTION: The use of alveolar recruitment maneuvers during general anaesthesia of horses is a potentially useful therapeutic option for the ventilatory management. While the routine application of recruitments would benefit from the availability of dedicated large animal ventilators their impact on ventilation and perfusion in the horse is not yet well documented nor completely understood. CASE HISTORY: A healthy 533 kg experimental horse underwent general anaesthesia in lateral recumbency. During intermittent positive pressure ventilation a stepwise alveolar recruitment maneuver was performed. MANAGEMENT: Anaesthesia was induced with ketamine and midazolam and maintained with isoflurane in oxygen using a large animal circle system. Mechanical ventilation was applied in pressure ventilation mode and an alveolar recruitment maneuver performed employing a sequence of ascending and descending positive end expiratory pressures. Next to the standard monitoring, which included spirometry, additionally three non-invasive monitoring techniques were used: electrical impedance tomography (EIT), volumetric capnography and respiratory ultrasonic plethysmography. The functional images continuously delivered by EIT initially showed markedly reduced ventilation in the dependent lung and allowed on-line monitoring of the dynamic changes in the distribution of ventilation during the recruitment maneuver. Furthermore, continuous monitoring of compliance, dead space fraction, tidal volumes and changes in end expiratory lung volume were possible without technical difficulties. FOLLOW: up The horse made an unremarkable recovery. CONCLUSION: The novel non-invasive monitoring technologies used in this study provided unprecedented insights into the physiology of lung collapse and recruitment. The synergic information of these techniques holds promise to be useful when developing and evaluating new ventilatory strategies in horses.

Interspecific somatic hybrids Solanum villosum (+) S. tuberosum, resistant to Phytophthora infestans.

The interspecific somatic hybrids 4x S. villosum (+) 2x S. tuberosum clone DG 81-68 (VT hybrids) were obtained and characterized molecularly and cytogenetically. The morphology of fusion-derived plants was intermediate in relation to the parental species. The expected ploidy level of the regenerants was 6x for the VT hybrids, but the real ploidy of the hybrids varied, with some of them being euploids, and others - aneuploids. The hybridity of the regenerants was verified by random amplified polymorphic DNA (RAPD) analysis. Despite the variation in ploidy, the RAPD patterns of the hybrids were mostly uniform, suggesting similarity of the genotypes of the VT clones. Genomic in situ hybridization (GISH) analysis discriminated between the chromosomes of both parental genomes in VT somatic hybrids and also confirmed their hybridity. The resistance of VT somatic hybrids to Phytophthora infestans was evaluated and all of the hybrids proved to be highly resistant. In search of the mechanisms involved in resistance of the Solanum species to P. infestans, the biochemical reactions occurring early after elicitor treatment were studied. The production of reactive oxygen species (ROS), as one of the earliest reactions induced by pathogens or their elicitors, was examined in the resistant wild species S. villosum, susceptible S. tuberosum clone DG 81-68 and in the VT hybrid, resistant to P. infestans. After treatment of the leaves with elicitor, the relative increase in ROS production was higher in leaves of the susceptible potato clone than in the resistant plants of S. villosum and the somatic hybrid.

Allometric scaling: analysis of LD50 data.

The need to identify toxicologically equivalent doses across different species is a major issue in toxicology and risk assessment. In this article, we investigate interspecies scaling based on the allometric equation applied to the single, oral LD (50) data previously analyzed by Rhomberg and Wolff. We focus on the statistical approach, namely, regression analysis of the mentioned data. In contrast to Rhomberg and Wolff's analysis of species pairs, we perform an overall analysis based on the whole data set. From our study it follows that if one assumes one single scaling rule for all species and substances in the data set, then ß = 1 is the most natural choice among a set of candidates known in the literature. In fact, we obtain quite narrow confidence intervals for this parameter. However, the estimate of the variance in the model is relatively high, resulting in rather wide prediction intervals.

Changes in the initial phase of lipid peroxidation induced by elicitor from Phytophthora infestans in Solanum species.

The initial phase of the lipid peroxidation process in leaves of Solanum nigrum var. gigantea, Solanum tuberosum cv Bzura and clone H-8105, which represent non-host resistance, field resistance and susceptibility, respectively, against Phytophthora infestans, was investigated. Based on quantitative and qualitative high-performance liquid chromatography (HPLC) analyses of free and esterified fatty acid hydroperoxides (FAHs), we characterized the lipid peroxidation process induced by the pathogen-derived elicitor, culture filtrate (CF), in leaves of the studied genotypes. In all plants, FAHs generated due to 13-lipoxygenase (LOX) action dominated over those from the non-enzymatic pathway. The FAHs derived from 9-LOX activity were found only in CF-treated leaves of the non-host resistant S. nigrum. However, experiments in vitro and in planta with exogenous linoleic acid (LA) as a substrate for LOX revealed high constitutive activity of 9-LOX in all genotypes, which increased in response to CF treatment. The time course changes in polyunsaturated fatty acid (PUFA) pools in the total lipid fractions as well as the degree of their oxidation suggested that CF-induced PUFA peroxidation was enhanced mostly in S. nigrum, less so in Bzura and least in the susceptible clone H-8105. The obtained results are discussed in light of the overall biochemical cell status of plants in the studied interactions.

Changes in the antioxidant status in leaves of Solanum species in response to elicitor from Phytophthora infestans.

Three Solanum genotypes with various polygenic resistance levels to the oomycete pathogen Phytophthora infestans (Mont.) De Bary were studied for their antioxidant response to the pathogen culture filtrate (CF). Detached plant leaves were treated with CF for 6, 18 and 30 h, and assayed for changes in hydrogen peroxide content, total ascorbate and glutathione pools and redox ratios (reduced form to total pool), as well as for changes in activities of ascorbate peroxidase, glutathione reductase and glutathione-S-transferase. In CF treated leaves of non-host resistant S. nigrum var. gigantea and field resistant S. tuberosum cv Bzura, the H(2)O(2) content did not change in comparison to water treated control leaves, whereas in the susceptible S. tuberosum clone H-8105 it decreased below the control level. In CF treated leaves of all genotypes, the total ascorbate pools were relatively unaltered and their redox ratio changed only transiently. In Bzura leaves the total glutathione content increased earlier than in the two other genotypes. The glutathione redox ratio remained rather stable, except for the susceptible clone H-8105, where it decreased transiently by about 42%. The relative increases in activity of all the studied enzymes were the highest in the susceptible clone H-8105. The results are discussed in the light of oxidative processes occurring in CF treated leaves. We conclude that stringent control of pro- and anti-oxidant reactions bringing the H(2)O(2) and/or cellular redox state to the threshold level is decisive for deployment of an effective defense strategy.

The upstream oxylipin profile of Arabidopsis thaliana: a tool to scan for oxidative stresses.

Various physiological imbalances lead to reactive oxygen species (ROS) overproduction and/or increases in lipoxygenase (LOX) activities, both events ending in lipid peroxidation of polyunsaturated fatty acids (PUFAs). Besides the quantification of such a process, the development of tools is necessary in order to allow the identification of the primary cause of its development and localization. A biochemical method assessing 9 LOX, 13 LOX and ROS-mediated peroxidation of membrane-bound and free PUFAs has been improved. The assay is based on the analysis of hydroxy fatty acids derived from PUFA hydroperoxides by both the straight and chiral phase high-performance liquid chromatography. Besides the upstream products of peroxidation of the 18:2 and 18:3 PUFAs, products coming from the 16:3 were characterized and their steady-state level quantified. Moreover, the observation that the relative amounts of the ROS-mediated peroxidation isomers of 18:3 were constant in leaves allowed us to circumvent the chiral analyses for the discrimination and quantification of 9 LOX, 13 LOX and ROS-mediated processes in routine experiments. The methodology has been successfully applied to decipher lipid peroxidation in Arabidopsis leaves submitted to biotic and abiotic stresses. We provide evidence of the relative timing of enzymatic and non-enzymatic lipid peroxidation processes. The 13 LOX pathway is activated early whatever the nature of the stress, leading to the peroxidation of chloroplast lipids. Under cadmium stress, the 9 LOX pathway added to the 13 LOX one. ROS-mediated peroxidation was mainly driven by light and always appeared as a late process.

The elicitor-induced oxidative processes in leaves of Solanum species with differential polygenic resistance to Phytophthora infestans.

Previous studies have shown that suspension-cultured cells of Solanum genotypes with various polygenic resistances to Phytophthora infestans differed in activities of early oxidative processes in response to culture filtrate (CF) from this pathogen. These studies have now been extended by analysing production of reactive oxygen species (ROS), lipid peroxidation and Lipoxygenase (LOX, E.C.1.13.11.12) activity induced by CF in detached leaves of S. tuberosum cv Bzura and clone H-8105, polygenically resistant and susceptible, respectively, as well as S. nigrum, nonhost, completely resistant. The relative increase in the ROS production was higher in the susceptible clone H-8105 than in both resistant genotypes. Lipid peroxidation increased only in the nonhost S. nigrum. An increase in lipid peroxidation in S. nigrum leaves coincided with enhanced LOX activity. In both S. tuberosum genotypes, significant increases in LOX activity were delayed and unaccompanied by changes in the level of lipid peroxidation. LOX activity attained a higher level in both of the resistant genotypes than in the susceptible one. The present results suggest that the involvement of both ROS production and LOX activity in the defense strategy in Solanum species/P. infestans interactions.

Plastoquinone: possible involvement in plant disease resistance.

The plant Solanum nigrum treated with the pathogen Phytophthora infestans-derived elicitor responded by elevated reactive oxygen species (ROS) production, lipid peroxidation and lipoxygenase (EC 1.13.11.12) activity in comparison with control plants indicating that oxidative stress took place. We demonstrate that these events are accompanied by a significant increase in plastoquinone (PQ) level. It is postulated that PQ may be associated with mechanisms maintaining a tightly controlled balance between the accumulation of ROS and antioxidant activity that determines the full expression of effective defence.