Directional asymmetry and direction-giving factors: Lessons from flowers with complex symmetry.

Directional asymmetry is a systematic difference between the left and right sides for structures with bilateral symmetry or a systematic differentiation among repeated parts for complex symmetry. This study explores factors that produce directional asymmetry in the flower of Iris pumila, a structure with complex symmetry that makes it possible to investigate multiple such factors simultaneously. The shapes and sizes of three types of floral organs, the falls, standards, and style branches, were quantified using the methods of geometric morphometrics. For each flower, this study recorded the compass orientations of floral organs as well as their anatomical orientations relative to the two spathes subtending each flower. To characterize directional asymmetry at the whole-flower level, differences in the average sizes and shapes according to compass orientation and relative orientation were computed, and the left-right asymmetry was also evaluated for each individual organ. No size or shape differences within flowers were found in relation to anatomical position; this may relate to the terminal position of flowers in Iris pumila, suggesting that there may be no adaxial-abaxial polarity, which is very prominent in many other taxa. There was clear directional asymmetry of shape in relation to compass orientation, presumably driven by a consistent environmental gradient such as solar irradiance. There was also clear directional asymmetry between left and right halves of every floral organ, most likely related to the arrangement of organs in the bud. These findings indicate that different factors are acting to produce directional asymmetry at different levels. In conventional analyses not recording flower orientations, these effects would be impossible to disentangle from each other and would probably be included as part of fluctuating asymmetry.

Correction to: Trastuzumab-induced upregulation of a protein set in extracellular vesicles emitted by ErbB2-positive breast cancer cells correlates with their trastuzumab sensitivity.

An amendment to this paper has been published and can be accessed via the original article.

Trastuzumab-induced upregulation of a protein set in extracellular vesicles emitted by ErbB2-positive breast cancer cells correlates with their trastuzumab sensitivity.

BACKGROUND: ErbB2/HER2 oncoprotein often drives breast cancers (BCs) which are treated with the anti-ErbB2 antibody trastuzumab. The efficacy of trastuzumab-based metastatic BC therapies is routinely assessed by imaging studies. Trastuzumab typically becomes ineffective in the case of this disease and is then replaced by other drugs. Biomarkers of BC trastuzumab response could allow imaging studies and the switch to other drugs to occur earlier than is now possible. Moreover, bone-only BC metastases can be hard to measure, and biomarkers of their trastuzumab response could facilitate further treatment decisions. Such biomarkers are presently unavailable. In this study, we searched for proteins whose levels in BC cell-emitted extracellular vesicles (EVs) potentially correlate with BC trastuzumab sensitivity. METHODS: We isolated EVs from cultured trastuzumab-sensitive and trastuzumab-resistant human BC cells before and after trastuzumab treatment and characterized these EVs by nanoparticle tracking analysis and electron microscopy. We found previously that ErbB2 drives BC by downregulating a pro-apoptotic protein PERP. We now tested whether trastuzumab-induced PERP upregulation in EVs emitted by cultured human BC cells correlates with their trastuzumab sensitivity. We also used mass spectrometry to search for additional proteins whose levels in such EVs reflect BC cell trastuzumab sensitivity. Once we identified proteins whose EV levels correlate with this sensitivity in culture, we explored the feasibility of testing whether their levels in the blood EVs of trastuzumab-treated metastatic BC patients correlate with patients' response to trastuzumab-based treatments. RESULTS: We found that neither trastuzumab nor acquisition of trastuzumab resistance by BC cells affects the size or morphology of EVs emitted by cultured BC cells. We established that EV levels of proteins PERP, GNAS2, GNA13, ITB1, and RAB10 correlate with BC cell trastuzumab response. Moreover, these proteins were upregulated during trastuzumab-based therapies in the blood EVs of a pilot cohort of metastatic BC patients that benefited from these therapies but not in those derived from patients that failed such treatments. CONCLUSIONS: Upregulation of a protein set in EVs derived from cultured breast tumor cells correlates with tumor cell trastuzumab sensitivity. It is feasible to further evaluate these proteins as biomarkers of metastatic BC trastuzumab response.

Value of Multidetector Computed Tomography in the Assessment of Achalasia Subtypes and Detection of Pulmonary and Thoracic Complications.

OBJECTIVE: To evaluate multidetector computed tomography (MDCT) findings in patients with achalasia, to assess its role in differentiating subtypes in detecting lung involvement and extra-esophageal thoracic complications. SUBJECTS AND METHODS: This clinical retrospective study included 51 patients with manometrically confirmed achalasia who underwent chest X-ray and MDCT in diagnostic work-up. Esophageal wall thickness and morphology, luminal dilatation, lung changes, and extra-esophageal manifestations were analyzed on MDCT by 2 readers. Wilcoxon, Kruskal-Wallis and Mann-Whitney test were used for assessing the differences among the achalasia subtypes, and intra-class correlation coefficients (ICC) assessing the inter-observer agreement between the measurements of 2 readers. RESULTS: Fourteen (27.5%) patients had achalasia subtype I, 21 (60.8%) had subtype II while 6 (11.8%) had subtype III. Esophageal wall thickness of the esophageal body (EB) and distal esophageal segment (DES) as well as nodular/lobulated appearance of DES were found significantly more often in subtype III (p = 0.024, p < 0.001, p = 0.009, respectively). Esophageal dilatation gradually decreased from subtype I to III (p = 0.006). Chest X-ray revealed lung changes in 9 (17%) and MDCT in 21 (41%) patients (p = 0.001), most frequently in subtype I, with predominance of ground-glass opacities. Tracheal/carinal compression was detected in 27 (52.9%) and left atrial compression in 17 (33.3%) patients. Excellent inter-observer agreement was observed in measuring the EB and DES wall thickness, and diameter of EB (ICC 0.829, 0.901, and 0.922). CONCLUSION: MDCT is a useful tool for detecting lung and extra-esophageal thoracic complications in patients with achalasia, and could be a valuable additional imaging modality in the differentiation of achalasia subtypes.

Phenotypic plasticity in response to environmental heterogeneity contributes to fluctuating asymmetry in plants: first empirical evidence.

Fluctuating asymmetry (FA) is widely used to quantify developmental instability (DI) in ecological and evolutionary studies. It has long been recognized that FA may not exclusively originate from DI for sessile organisms such as plants, because phenotypic plasticity in response to heterogeneities in the environment might also produce FA. This study provides the first empirical evidence for this hypothesis. We reasoned that solar irradiance, which is greater on the southern side than on the northern side of plants growing in the temperate zone of the Northern Hemisphere, would cause systematic morphological differences and asymmetry associated with the orientation of plant parts. We used geometric morphometrics to characterize the size and shape of flower parts in Iris pumila grown in a common garden. The size of floral organs was not significantly affected by orientation. Shape and particularly its asymmetric component differed significantly according to orientation for three different floral parts. Orientation accounted for 10.4% of the total shape asymmetry within flowers in the falls, for 11.4% in the standards and for 2.2% in the style branches. This indicates that phenotypic plasticity in response to a directed environmental factor, most likely solar irradiance, contributes to FA of flowers under natural conditions. That FA partly results from phenotypic plasticity and not just from DI needs to be considered by studies of FA in plants and other sessile organisms.

Insights Into Roles for Health-Care Professionals in Meeting the Needs of Older Adults and Unpaid Caregivers During Health-Care Transitions.

We provided insights from older adults, their unpaid caregivers, and health-care professionals into specific roles for professionals within the health system to better meet the needs of community-dwelling older adults and their unpaid caregivers experiencing transitions between health services. We used a qualitative approach to collect data within one Canadian province from older adults and unpaid caregivers of older adults who participated in focus groups ( n = 98) and professionals working in the health system who participated in an online survey ( n = 52). Questions included experiences with health service transitions, strengths, challenges, and suggestions to improve transitions. Thematic analysis resulted in identifying seven specific roles for professionals in supporting health-care transitions: information and education, planning for future health needs, supporting the acceptance of necessary care, facilitating access to the right services at the right time, facilitating communication between services, facilitating the discharge planning process and advocacy for older adults and unpaid caregivers. Our results based on evidence from older adults, unpaid caregivers, and health-care professionals will inform future research and further development of the instrumental and relational roles for professionals supporting older adults and their caregivers experiencing health-care transitions.

Variability of Silver Fir Needle (Abies alba Mill.) Anatomical Features in the Southeast Europe Natural Populations.

The survival of marginal/peripheral silver fir (Abies alba Mill.) populations in the broader region of Southeast Europe is endangered due to climate change and population decline. This study aimed to determine the level and pattern of variability for the anatomical traits of needles and the possibility of linking the pattern of phenotypic variability with environmental factors. In most of the analyzed needle traits, the statistically significant variability between populations was determined. According to the results of the multivariate principal component analysis, it is evident that the populations are distinct from each other, in three groups. The climatic factors Hargreaves reference evaporation, mean annual temperature, and growing degree-days, were statistically significantly correlated. The altitude and heating degree-days are statistically significantly correlated with the following three environmental factors: Hargreaves reference evaporation, mean annual temperature and growing degree-days, but negatively with others. The paper's findings indicate significant moderate and high correlations between the anatomical traits of the needles' central bundle diameter with the resin duct diameter, the distance between the vascular bundle and the resin duct and the epidermis thickness with cuticle, the resin duct diameter with the distance between the vascular bundle and the resin duct and the epidermis thickness with cuticle, as well as the distance between the vascular bundle and the resin duct with the hypodermis height and the epidermis thickness with cuticle. The results of agglomerative hierarchical clustering analysis, performed for anatomical and climatic traits, confirmed the existence of three groups of tested populations according to the altitude gradient. Research results provide knowledge on the diversity and structure of Abies alba populations of Southeast Europe, important for further research and guidelines for the species' conservation and genetic variability preservation in the southern marginal distribution area and keeping in line with climate change projections.

Predicting the Responses of Functional Leaf Traits to Global Warming: An In Situ Temperature Manipulation Design Using Iris pumila L.

Phenotypic plasticity is widely acknowledged as one of the most common solutions for coping with novel environmental conditions following climate change. However, it is less known whether the current amounts of trait plasticity, which is sufficient for matching with the contemporary climate, will be adequate when global temperatures exceed historical levels. We addressed this issue by exploring the responses of functional and structural leaf traits in Iris pumila clonal individuals to experimentally increased temperatures (~1.5 °C) using an open top chamber (OTC) design. We determined the phenotypic values of the specific leaf area, leaf dry matter content, specific leaf water content, and leaf thickness in the leaves sampled from the same clone inside and outside of the OTC deployed on it, over seasons and years within two natural populations. We analyzed the data using a repeated multivariate analysis of variance, which primarily focusses on the profiles (reaction norms (RNs)) of a variable gathered from the same individual at several different time points. We found that the mean RNs of all analyzed traits were parallel regardless of experienced temperatures, but differed in the level and the shape. The populations RNs were similar as well. As the amount of plasticity in the analyzed leaf trait was adequate for coping with elevated temperatures inside the OTCs, we predict that it will be also sufficient for responding to increased temperatures if they exceed the 1.5 °C target.

Is COVID-19 another case of the obesity paradox? Results from an international ecological study on behalf of the REPROGRAM Consortium Obesity study group.

Introduction: Obesity has emerged as one of the major risk factors of severe morbidity and cause-specific mortality among severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infected individuals. Patients with obesity also have overlapping cardiovascular diseases and diabetes, which make them increasingly vulnerable. This novel ecological study examines the impact of obesity and/or body mass index (BMI) on rates of population-adjusted cases and deaths due to coronavirus disease 2019 (COVID-19). Material and methods: Publicly available datasets were used to obtain relevant data on COVID-19, obesity and ecological variables. Group-wise comparisons and multivariate logistic regression analyses were performed. The receiver operating characteristic curve (ROC) was plotted to compute the area under the curve. Results: We found that male BMI is an independent predictor of cause-specific (COVID-19) mortality, and not of the caseload per million population. Countries with obesity rates of 20-30% had a significantly higher (approximately double) number of deaths per million population to both those in < 20% and > 30% slabs. We postulate that there may be a U-shaped paradoxical relationship between obesity and COVID-19 with the cause-specific mortality burden more pronounced in the countries with 20-30% obesity rates. These findings are novel along with the methodological approach of doing ecological analyses on country-wide data from publicly available sources. Conclusions: We anticipate, in light of our findings, that appropriate targeted public health approaches or campaigns could be developed to minimize the risk and cause-specific morbidity burden due to COVID-19 in countries with nationwide obesity rates of 20-30%.

Percutaneous Computed Tomography-Guided Oxygen-Ozone (O2O3) Injection Therapy in Patients with Lower Back Pain-An Interventional Two-Year Follow-Up Study of 321 Patients.

OBJECTIVES: To assess the effect of oxygen-ozone therapy guided by percutaneous Computed Tomography (CT) compared to corticosteroids in individuals experiencing lower back pain (LBP) not attributed to underlying bone-related issues. METHODS: A total of 321 patients (192 males and 129 females, mean age: 51.5 ± 15.1 years) with LBP were assigned to three treatment groups: group A) oxygen-ozone only, group B) corticosteroids only, group C) oxygen-ozone and corticosteroids. Treatment was administered via CT-guided injections to the intervertebral disc (i.e., intradiscal location). Clinical improvement of pain and functionality was assessed via self-reported pain scales and magnetic resonance (MR) and CT imaging. RESULTS: At all follow-up times, the mean score of the numeric rating scale and the total global pain scale (GPS) of study groups receiving oxygen-ozone (groups A and C) were statistically significantly lower than the study group receiving corticosteroids only (group B), with p < 0.001. There was a statistically significant difference between groups A and C at 30 days for the numeric rating scale. CONCLUSIONS: The percutaneous application of oxygen-ozone in patients with LBP due to degeneration of the lumbosacral spine showed long-lasting significant pain reduction of up to two years post-treatment when compared to corticosteroids alone. Combination therapy of oxygen-ozone and corticosteroids can be useful as corticosteroids showed statistically significant improvement in LBP earlier than the oxygen-ozone-only treatment.

Assessment of Sex-Specific Toxicity and Physiological Responses to Thymol in a Common Bean Pest Acanthoscelides obtectus Say.

Acanthoscelides obtectus Say (Coleoptera: Chrysomelidae: Bruchinae), is one of the most important pests of the common bean Phaseolus vulgaris L. Without appropriate management it may cause significant seed loss in storages. In search for means of environmentally safe and effective protection of beans we assessed biological activity of thymol, an oxygenated monoterpene present in essential oils of many aromatic plants. We studied contact toxicity of thymol on bean seeds and its effects on adult longevity and emergence in F1 generation. Furthermore, we determined acetylcholinesterase (AChE), superoxide dismutase (SOD), catalase (CAT), mixed-function oxidase (MFO), carboxylesterases (CarE) and glutathione S-transferase (GST) activities in response to 24 h exposure of beetles to sublethal and lethal thymol concentrations. Our results showed that thymol decreased adult survival, longevity and percentage of adult emergence. Higher median lethal concentration (LC50) was recorded in females indicating their higher tolerance comparing to males. Overall, activities of SOD, CAT and CarE increased at sublethal and MFO increased at both sublethal and lethal thymol concentrations. On the other hand, GST and AChE activities decreased along with the increase in thymol concentrations from sublethal (1/5 of LC50, 1/2 of LC50) to lethal (LC50). Enzyme responses to the presence of thymol on bean seed were sex-specific. In the control group females had lower CarE and higher SOD, CAT and GST activity than males. In treatment groups, females had much higher CAT activity and much lower CarE activity than males. Our results contribute to deeper understanding of physiological mechanisms underlying thymol toxicity and tolerance which should be taken into account in future formulation of a thymol-based insecticide.

Validation of endogenous controls for gene expression studies in peripheral lymphocytes from war veterans with and without PTSD.

BACKGROUND: Selection of appropriate endogenous control is a critical step in gene expression analysis. The aim of this study was to evaluate expression stability of four frequently used endogenous controls: beta-actin, glyceraldehyde-3-phosphate dehydrogenase, beta2-microglobulin and RNA polymerase II polypeptide A in peripheral blood mononuclear cells from war veterans with and without posttraumatic stress disorder (PTSD). The study was designed as to identify suitable reference gene(s) for normalization of gene expression in peripheral blood mononuclear cells in response to war trauma and/or PTSD. RESULTS: The variability in expression of the four endogenous controls was assessed by TaqMan Real-time RT-PCR in peripheral blood mononuclear cells from: war veterans with current PTSD, those with lifetime PTSD, trauma controls and healthy subjects. Expression stability was analyzed by GeNorm and NormFinder software packages, and by direct comparison of Ct values. Both, GeNorm and NormFinder identified beta-actin and glyceraldehyde-3-phosphate dehydrogenase as a pair of genes with the lowest stability value. CONCLUSIONS: The combination of beta-actin and glyceraldehyde-3-phosphate dehydrogenase appeared to be the most suitable reference for studying alterations in gene expression in peripheral blood mononuclear cells related to vulnerability and resilience to PTSD, as well as to trauma-provoked developing of this disorder and recovery from it. Using glyceraldehyde-3-phosphate dehydrogenase, beta-actin and beta2-microglobulin as individual endogenous controls would provide satisfactory data, while RNA polymerase II polypeptide A could not be recommended.

Toxic, Oviposition Deterrent and Oxidative Stress Effects of Thymus vulgaris Essential Oil against Acanthoscelides obtectus.

The bean weevil Acanthoscelides obtectus Say (Coleoptera: Chrysomelidae: Bruchinae) can cause significant losses in production of its primary host common bean Phaseolus vulgaris L. To avoid bean protection with environmentally risky chemical insecticides and provide sustainable and safe production of food, new pest management methods based on natural compounds are investigated. In the present study, we evaluated protective potential of the essential oil (EO) from the common thyme Thymus vulgaris L. applied on bean seeds. We assessed residual contact toxicity of thyme EO and its effects on A. obtectus longevity, oviposition and adult emergence. Furthermore, to elucidate the role of oxidative stress in thyme EO toxicity, we estimated the levels of oxidatively damaged proteins and lipids, as well as the level of thiols which have important role for antioxidant capacity. We found that thyme oil significantly reduced adult survival and longevity, induced oxidative damage to lipids and proteins and depleted protein and non-protein thiols in a concentration-dependent manner. Females appeared to be more tolerant to thyme oil treatment than males. Sublethal EO concentrations affected oxidative stress indices, deterred oviposition and strongly inhibited adult emergence. The results suggest that thyme oil has the potential to be used as an ecofriendly insecticide for A. obtectus control.

Influence of Syrgical Trauma on Nitric Oxide and Nitrotyrosine Serum Levels in Patients Undergoing Laparoscopic Or Conventional Cholecystectomy.

BACKGROUND: Oxidative stress represents tissue damage caused by reactive forms of oxygen and nitrogen due to the inability of antioxidant mechanisms to reduce reactive forms into more stable ones. The aim of the study was to evaluate the influence of surgical trauma on nitric oxide (NO) and nitrotyrosine (NT) values in patients undergoing conventional and laparoscopic cholecystectomy. METHODS: A prospective study included sixty patients from the Department of Emergency Surgery, Clinical Centre of Serbia who were operated for gallstone related chronic cholecystitis. All the patients enrolled in the study underwent cholecystectomy; the first group was operated conventionally (30 patients - control group), while the second group was operated laparoscopically (30 patients - treatment group). RESULTS: There were no statistically significant differences in the values of NO and its postoperative changes in both groups, the conventionally operated group (p=0.943) and the laparoscopically operated group (p=0.393). We found an increase in NT values 24 hours postoperatively (p=0.000) in the conventionally operated patients, while in the group operated laparoscopically we didn't find statistically significant changes in the values of NT (conventionally operated group (p=0.943) and laparoscopically operated group (p=0.393)). CONCLUSIONS: In our study, we found a significant increase in NT values 24 hours postoperatively in conventionally operated patients i.e. the control group, vs. the treatment group. Further randomized studies are needed for a better understanding of the impact of surgical trauma on oxidative stress response.

Adaptive flexibility of enzymatic antioxidants SOD, APX and CAT to high light stress: The clonal perennial monocot Iris pumila as a study case.

High solar radiation has been recognized as one of the main causes of the overproduction of reactive oxygen species (ROS) and oxidative stress in plants. To remove the excess of ROS, plants use different antioxidants and tune their activity and/or isoform number as required for given light conditions. In this study, the adaptiveness of light-induced variation in the activities and isoform patterns of key enzymatic antioxidants SOD, APX and CAT was tested in leaves of Iris pumila clonal plants from two natural populations inhabiting a sun exposed dune site and a forest understory, using a reciprocal-transplant experiment. At the exposed habitat, the mean enzymatic activity of total SODs was significantly greater than that in the shaded one, while the amount of the mitochondrial MnSOD was notably higher compared to the plastidic Cu/ZnSOD. However, the number of Cu/ZnSOD isoforms was greater in the forest understory relative to the exposed site (three vs. two, respectively). An inverse relationship recorded between the quantities of MnSOD and Cu/ZnSOD in alternative light habitats might indicate that the two enzymes compensate each other in maintaining intracellular ROS and redox balance. The adaptive population differentiation in APX activity was exclusively recorded in the open habitat, which indicated that the synergistic effect of high light and temperature stress could be the principal selective factor, rather than high light alone. The enzymatic activity of CAT was similar between the two populations, implicating APX as the primary H2O2 scavenger in the I. pumila leaves exposed to high light intensity.

Mineralocorticoid receptor and heat shock protein expression levels in peripheral lymphocytes from war trauma-exposed men with and without PTSD.

Alterations in the number and functional status of mineralocorticoid (MR) and glucocorticoid receptors (GR) may contribute to vulnerability to posttraumatic stress disorder (PTSD). Corticosteroid receptors are chaperoned by heat shock proteins Hsp90 and Hsp70. We examined relations between corticosteroid receptor and heat shock protein expression levels, and related them with war trauma exposure, PTSD and resilience to PTSD. Relative levels of MR, Hsp90 and Hsp70 were determined by immunoblotting in lymphocytes from war trauma-exposed men with current PTSD (current PTSD group, n=113), with life-time PTSD (life-time PTSD group, n=61) and without PTSD (trauma control group, n=88), and from non-traumatized healthy controls (healthy control group, n=85). Between-group differences in MR, Hsp90 and Hsp70 levels and in MR/GR ratio were not observed. The level of MR was correlated with both Hsp90 and Hsp70 levels in trauma control and healthy control groups. On the other hand, GR level was correlated only with Hsp90 level, and this correlation was evident in current PTSD and trauma control groups. In conclusion, PTSD and exposure to trauma are not related to changes in lymphocyte MR, Hsp90 or Hsp70 levels, but may be associated with disturbances in corticosteroid receptors interaction with heat shock proteins.

Lymphocyte glucocorticoid receptor expression level and hormone-binding properties differ between war trauma-exposed men with and without PTSD.

OBJECTIVE: Posttraumatic stress disorder (PTSD) has been shown to be associated with altered glucocorticoid receptor (GR) activity. We studied the expression and functional properties of the receptor in peripheral blood mononuclear cells (PBMCs) from non-traumatized healthy individuals (healthy controls; n=85), and war trauma-exposed individuals with current PTSD (n=113), with life-time PTSD (n=61) and without PTSD (trauma controls; n=88). The aim of the study was to distinguish the receptor alterations related to PTSD from those related to trauma itself or to resilience to PTSD. METHODS: Functional status of the receptor was assessed by radioligand binding and lysozyme synthesis inhibition assays. The level of GR gene expression was measured by quantitative PCR and immunoblotting. RESULTS: Current PTSD patients had the lowest, while trauma controls had the highest number of glucocorticoid binding sites (Bmax) in PBMCs. Hormone-binding potential (Bmax/KD ratio) of the receptor was diminished in the current PTSD group in comparison to all other study groups. Correlation between Bmax and KD that normally exists in healthy individuals was decreased in the current PTSD group. Contrasting Bmax data, GR protein level was lower in trauma controls than in participants with current or life-time PTSD. CONCLUSIONS: Current PTSD is characterized by reduced lymphocyte GR hormone-binding potential and by disturbed compensation between Bmax and hormone-binding affinity. Resilience to PTSD is associated with enlarged fraction of the receptor molecules capable of hormone binding, within the total receptor molecule population in PBMCs.