Chronic impacts of invasive herbivores on a foundational forest species: a whole-tree perspective.

Forests make up a large portion of terrestrial plant biomass, and the long-lived woody plants that dominate them possess an array of traits that deter consumption by forest pests. Although often extremely effective against native consumers, invasive species that avoid or overcome these defenses can wreak havoc on trees and surrounding ecosystems. This is especially true when multiple invasive species co-occur, since interactions between invasive herbivores may yield non-additive effects on the host. While the threat posed by invasive forest pests is well known, long-term field experiments are necessary to explore these consumer-host interactions at appropriate spatial and temporal scales. Moreover, it is important to measure multiple variables to get a "whole-plant" picture of their combined impact. We report the results of a 4-yr field experiment addressing the individual and combined impacts of two invasive herbivores, the hemlock woolly adelgid (Adelges tsugae) and elongate hemlock scale (Fiorinia externa), on native eastern hemlock (Tsuga canadensis) in southern New England. In 2011, we planted 200 hemlock saplings into a temperate forest understory and experimentally manipulated the presence/absence of both herbivore species; in 2015, we harvested the 88 remaining saplings and assessed plant physiology, growth, and resource allocation. Adelgids strongly affected hemlock growth: infested saplings had lower above/belowground biomass ratios, more needle loss, and produced fewer new needles than control saplings. Hemlock scale did not alter plant biomass allocation or growth, and its co-occurrence did not alter the impact of adelgid. While both adelgid and scale impacted the concentrations of primary metabolites, adelgid effects were more pronounced. Adelgid feeding simultaneously increased free amino acids local to feeding sites and a ~30% reduction in starch. The cumulative impact of adelgid-induced needle loss, manipulation of nitrogen pools, and the loss of stored resources likely accelerates host decline through disruption of homeostatic source-sink dynamics occurring at the whole-plant level. Our research stresses the importance of considering long-term impacts to predict how plants will cope with contemporary pressures experienced in disturbed forests.

Directional K+ channel insertion in a single phospholipid bilayer: Neutron reflectometry and electrophysiology in the joint exploration of a model membrane functional platform.

We investigated the insertion of small potassium (K+) channel proteins (KcvMA-1D and KcvNTS) into model membranes and the lipid-protein structural interference, combining neutron reflectometry and electrophysiology. Neutron reflectometry experiments showed how the transverse structure and mechanical properties of the bilayer were modified, upon insertion of the proteins in single model-membranes, either supported on solid substrate or floating. Parallel electrophysiology experiments were performed on the same channels reconstituted in free-standing planar lipid bilayers, of both typical composition and matched to the neutron reflectometry experiment, assessing their electrical features. Functional and structural results converge in detecting that the proteins, conical in shape, insert with a directionality, cytosolic side first. Our work addresses the powerful combination of the two experimental approaches. We show here that membrane structure spectroscopy and ion channel electrophysiology can become synergistic tools in the analysis of structural-functional properties of biomimetic complex environment.

Defense response in non-genomic model species: methyl jasmonate exposure reveals the passion fruit leaves' ability to assemble a cocktail of functionally diversified Kunitz-type trypsin inhibitors and recruit two of them against papain.

MAIN CONCLUSION: Multiplicity of protease inhibitors induced by predators may increase the understanding of a plant's intelligent behavior toward environmental challenges. Information about defense mechanisms of non-genomic model plant passion fruit (Passiflora edulis Sims) in response to predator attack is still limited. Here, via biochemical approaches, we showed its flexibility to build-up a broad repertoire of potent Kunitz-type trypsin inhibitors (KTIs) in response to methyl jasmonate. Seven inhibitors (20-25 kDa) were purified from exposed leaves by chromatographic techniques. Interestingly, the KTIs possessed truncated Kunitz motif in their N-terminus and some of them also presented non-consensus residues. Gelatin-Native-PAGE established multiple isoforms for each inhibitor. Significant differences regarding inhibitors' activity toward trypsin and chymotrypsin were observed, indicating functional polymorphism. Despite its rarity, two of them also inhibited papain, and such bifunctionality suggests a recruiting process onto another mechanistic class of target protease (cysteine-type). All inhibitors acted strongly on midgut proteases from sugarcane borer, Diatraea saccharalis (a lepidopteran insect) while in vivo assays supported their insecticide properties. Moreover, the bifunctional inhibitors displayed activity toward midgut proteases from cowpea weevil, Callosobruchus maculatus (a coleopteran insect). Unexpectedly, all inhibitors were highly effective against midgut proteases from Aedes aegypti a dipteran insect (vector of neglected tropical diseases) opening new avenues for plant-derived PIs for vector control-oriented research. Our results reflect the KTIs' complexities in passion fruit which could be wisely exploited by influencing plant defense conditions. Therefore, the potential of passion fruit as source of bioactive compounds with diversified biotechnological application was strengthened.

Effects of Wolbachia elimination and B-vitamin supplementation on bed bug development and reproduction.

Obligate blood feeders, such as Cimex lectularius (common bed bug), have symbiotic associations with nutritional endosymbionts that produce B-vitamins. To quantify the symbiont's contribution to host fitness in these obligate mutualisms, the symbiont must be eliminated and its absence rigorously confirmed. We developed and validated procedures for complete elimination of Wolbachia (Wb) in bed bugs and quantified development and reproduction in bed bugs with and without Wb and with and without B-vitamins supplementation. Aposymbiotic bed bugs had slower nymphal development, reduced adult survivorship, smaller adult size, fewer eggs per female, and lower hatch rate than bed bugs that harbored Wb. In aposymbiotic bed bugs that were fed B-vitamins-supplemented blood, nymph development time, adult survivorship and hatch rate recovered to control levels, but adult size and egg number only partially recovered. These results underscore the nutritional dependence of bed bugs on their Wb symbiont and suggest that Wb may provide additional nutritional benefits beyond the B-vitamin mix that we investigated.

Life History and Rearing of Anastatus orientalis (Hymenoptera: Eupelmidae), an Egg Parasitoid of the Spotted Lanternfly (Hemiptera: Fulgoridae).

To support efforts to manage and contain spotted lanternfly (SLF), Lycorma delicatula White (Hemiptera: Fulgoridae), research is being conducted to develop classical biological control methods. To date, two potential biocontrol agents from China have been identified: an egg parasitoid, Anastatus orientalis, and a nymphal parasitoid, Dryinus sinicus Olmi (Hymenoptera: Dryinidae). The research detailed here focuses on investigating the biology and rearing of A. orientalis to assess its potential efficacy in a biocontrol program and optimize its rearing. Female wasps lived significantly longer than male wasps (68 and 23 d, respectively) and females produced an average of 94 total progeny that successfully emerged as adults, with most progeny produced between weeks one and four of the females' lives. The sex ratio of the progeny, with no re-mating, was initially highly female-biased but became progressively more male-biased, likely due to sperm depletion. There was no evidence of additional mortality to SLF eggs from wasp host feeding, but the data were highly variable and the sample size was small. There was high parasitoid emergence when oviposition conditions mimicked mid-September Beijing temperature and photoperiod; however, there was little emergence under 25°C and long-day conditions because most progeny entered a diapause. Storage of parasitized eggs in 5°C chill lowered parasitoid emergence rates. Lastly, there was no evidence that storing field-collected SLF egg masses in 5°C for 10 mo prior to parasitization affected parasitism rates. These findings inform our rearing protocol for A. orientalis and facilitate our testing of this species as a potential biological control agent for SLF.

Cardiac Biomarkers Release in Preadolescent Athletes After an High Intensity Exercise.

INTRODUCTION: An elevation of cardiac troponins has been described in healthy athletes after endurance exercises. The clinical significance of this increase is unclear and the lack of awareness of this phenomenon may lead to inappropriate management of these subjects. AIM: We sought to determine wether an intensive cycling training could determine a biomarkers elevation. METHODS: We evaluated serum high sensitivity cardiac troponin T, NH(2)-terminal pro-brain natriuretic peptide, CK-MB and CK in 21 healthy male preadolescent athletes (age 9.2 ± 1.7 years) after an intensive cycling training prolonged until muscular exhaustion (mean duration 16'41″). During exercise heart rhythm and rate were monitored with Holter. RESULTS: 62% of the group had an elevation of cardiac biomarkers: specifically, 6 children had an increase in troponin levels; 3 of them had an elevation of pro-brain natriuretic peptides as well. Pro-brain natriuretic peptides resulted increased in 9 subjects. There was no relation between troponin elevation and heart rate, age or exercise duration; subjects with increased pro-brain natriuretic peptides had mean and maximal heart rates lower than children with normal natriuretic peptides levels. Other sports were performed in 50% of subjects with normal troponins and only in 17% of those with increased values. CONCLUSIONS: A short, high-intensity exercise caused an elevation of cardiac biomarkers in 62% of our subjects. The grade of training may influence the release of troponin and this increase is probably related to a temporary discrepancy between O2 delivery and consumption. Increases in natriuretic peptides levels are possibly expression of different adaptations to exercise.

Which variable of stenosis severity best describes the significance of an isolated left anterior descending coronary artery lesion? Correlation between quantitative coronary angiography, intracoronary Doppler measurements and high dose dipyridamole echocardiography.

OBJECTIVES: This study sought to investigate the angiographic or intracoronary Doppler variables of stenosis severity that best correlate with the results of dipyridamole echocardiography. BACKGROUND: Quantitative coronary angiography and intracoronary Doppler flow velocity assessments are the commonly used techniques for the objective identification of significant coronary artery stenosis. METHODS: Thirty patients with an isolated lesion of the left anterior descending coronary artery (LAD) were studied by means of on-line quantitative coronary arteriography, intracoronary Doppler flow velocity measurements and dipyridamole echocardiography 6 months after percutaneous transluminal coronary angioplasty. The quantitative arteriographic analyses were performed on-line; post-stenotic Doppler flow velocities were measured at baseline and after adenosine infusion. Angiographic and Doppler measurements were compared with the corresponding dipyridamole echocardiographic data and analyzed by discriminant analysis. RESULTS: The dipyridamole echocardiographic response was positive in 11 patients (37%). The best cutoff values for predicting an abnormal echocardiographic response were 1) stenotic flow reserve of 2.8 (p = 0.0001); 2) 59% diameter stenosis (p = 0.0001); 3) minimal lumen diameter of 1.35 mm (p = 0.001); 4) coronary flow reserve of 2.0 (p = 0.0002); and 5) maximal peak velocity of 60 cm/s during hyperemia (p = 0.04). Multivariate analysis identified stenotic flow reserve as the only independent predictor of ischemia during dipyridamole echocardiography. CONCLUSIONS: Stenotic flow reserve is the variable that best describes the functional significance of an isolated LAD lesion, and a value of 2.8 is the best predictor of a positive dipyridamole echocardiographic response. Furthermore, angiographic variables of stenosis severity relate to echocardiographic test results better than intracoronary Doppler variables.

Whole exome sequencing reveals a novel de novo FOXC1 mutation in a patient with unrecognized Axenfeld-Rieger syndrome and glaucoma.

We report the identification of a novel mutation in the fork-head box C1 (FOXC1) gene which occurred de novo in an Italian patient with unrecognized Axenfeld-Rieger syndrome. He was previously diagnosed as having late recognized primary congenital glaucoma at the age of 14 years and was subsequently subjected to multiple surgical interventions due to uncontrolled intraocular pressure and progressive visual field loss. After exclusion of mutations in CYP1B1 and MYOC, trio-whole-exome sequencing revealed de novo in frame deletion in the coding region of the FOXC1 gene (c.407_409delGTC, p.V137del) leading to a deletion of the evolutionary conserved amino acid Valine at position 137 of the protein. Molecular modeling predicted that Val137 deletion impairs FOXC1 DNA-binding capacity and transcriptional activation. Since loss-of-function mutations in FOXC1 are associated with Axenfeld-Rieger syndrome, the genetic findings in combination with re-evaluation of the patient's clinical data resulted in a corrected diagnosis of Axenfeld-Rieger syndrome with developmental glaucoma. We therefore suggest that in addition to CYP1B1 and MYOC, FOXC1 should be included in the genetic analysis of cases with unclear glaucomatous phenotypes to ensure proper diagnosis, adequate treatment and appropriate genetic counseling.

Stent implantation without predilation in patients with a single, noncalcified coronary artery lesion.

The purpose of this study was to assess the feasibility of stent implantation without predilation in patients with a single, noncalcified coronary artery lesion. A total of 122 patients were randomized to receive a stent with or without predilation; direct stent placement was possible in 59 of the 61 patients (97%) with an immediate and long-term clinical follow-up similar to that observed in the group of patients treated conventionally.

Evaluation of endocrine ophthalmopathy with saccadic eye movements.

Horizontal saccades were examined in 25 patients with hyperthyroidism and/or endocrine ophthalmopathy (EOP) using the infrared reflection method. With one exception none had restriction of horizontal eye movements. Conventional saccadic parameters were usually normal. A standardized test for muscle fatigue, however, gave pathological results in all but one of the patients with EOP, and also in three of seven patients with hyperthyroidism but no clinical EOP. In one of the latter the oculographic abnormality disappeared with treatment, while another developed clinical EOP. Performing a saccadic fatigue test seems to be useful for detecting early EOP and especially for monitoring its course.

Isolated congenital left ventricular diverticulum.

Two cases of isolated left ventricular diverticulum are presented. A 12-year-old boy and a 7-year-old girl, both asymptomatic, were admitted to our Department for evaluation of an abnormal electrocardiogram and an abnormal cardiac silhouette, respectively. Both patients had the diagnosis confirmed by cardiac catheterization and angiography and underwent successful surgical correction of the abnormality. We discuss the diagnosis, prognosis, and surgical treatment of this entity and we advocate early surgical treatment for all diagnosed ventricular diverticula, even if asymptomatic.

Age-related frequency of viral lesions and their association with uterine cervical intraepithelial neoplasia.

Data collection forms of 50,165 routine smears were examined, and of these 47,577 were considered for this study. Viral cytological lesions (VL) were observed in 101 cases: of these, 19 were associated with cervical intraepithelial neoplasia. The total frequency of VL was 0.21%, with the peak in the teenager group (0.49%). The frequency of CIN among VL showed an overall frequency of 18.8%: this was null under 21 years of age and highest (50%) between 51 and 60 years of age. The frequency of VL in relation to clinical provenance of patients and the season of the year did not attain significance.

Connective tissue regeneration after laser-CO2 therapy.

The ultrastructural pattern of fibroblasts involved in the healing process of laser-treated tissues has been investigated. This study has been performed on the uterine cervix, the epithelial abnormalities of which are frequently treated by laser. Observations carried out immediately and two and seven days after treatment, have demonstrated that: fibroblasts show an active and almost uniform pattern; myofibroblasts are not present; and macrophages are limited in number. This healing process is interpreted as a proliferative pattern of the connective tissue which differs from that of some benign tumors and of healing wounds.

[Affective disorders in subcortical ischemic cerebrovascular pathology. Prospective clinical study of 43 patients]. / Trastornos afectivos en la patología vascular cerebral isquémica subcortical. Estudio clínico prospectivo en 43 pacientes.

To evaluate the prevalence and clinical features of affective disorders (AD) associated with subcortical ischemic cerebrovascular disease (mainly localized in the deep areas depending from the anterior, middle and posterior cerebral arteries), a prospective clinical study was carried out in 43 patients with lacunar infarctions (LI). It was found that AD were uncommon in these patients (25%) (1/43). The prevalences of depression in the major lacunar syndromes of the series were the following, from more to less common: pure motor hemiparesis (35%) (7/20); dysarthria-clumsy hand (25%) (1/4); pure sensorial syndrome (16.5%) (2/12), and atypical syndromes (no patient). The capsular topography was associated with AD in 32% (7/22); this association was more uncommon in thalamic (15%) (2/13) or pontine (25%) (1/4) areas. AD had a significant predominance (p = 0.027) in the lesions involving the dominant cerebral hemisphere (63.5%), mainly when these resulted in clinically mild or moderate/severe disability. Past history of AD was also significantly associated with depression (p = 0.0018). Our results show that affective disorders in lacunar infarctions are usually associated with: 1) the type of clinical syndrome; 2) the topography of the lesion; 3) the involved cerebral hemisphere; 4) the associate neurological focality, and 5) the previous occurrence of affective disorders.

[Non-rachitic hypophosphatemic osteopathy]. / Osteopatia ipofosfatemica non rachitica.

We have identified a patient with a condition called Hypophosphatemic Bone Disease (HBD). This disorder of phosphate metabolism is unrecorded by Dent in his final classification of metabolic bone diseases and has been described by Scriver C.R. and coll. Although the condition is in same ways analogous to X-Linked Hyophosphatemia (XLH), there are important differences between the two diseases. For example, there is selective impairment in the tubular reabsorption of phosphate in HBD but the defect is less severe and it is clearly different from that described in XLH. Clinical manifestations of HBD appear in infancy, but the dwarfism and the bone changes are less severe than in XLH at comparable concentrations of plasmatic phosphorus in the two diseases. While in both conditions there is osteomalacia of endostal trabecular bone, only in XLH is florid rickets present, affecting the epiphyses and compromising linear growth. The phosphaturic response to PTH infusion is abnormal in qualitative aspects, but it is present in HBD, and this differs considerably from that described in XLH. The treatment with oral load of phosphates and 1,25(OH)2D3 in every patient with HBD, and so in our patient, is accompanied by increase in serum phosphorus, with improved tubular reabsorption of phosphate anion and a fall of hydrossiprolinuria with bone healing; this combination of responses is not present in XLH.

[Immediate and late arrhythmia in patients operated on for tetralogy of Fallot]. / Aritmie immediate e a lungo termine nei pazienti operati per tetralogia di Fallot.

UNLABELLED: Arrhythmias are a frequent complication after repair of tetralogy of Fallot (TOF). We present our experience with 97 patients with special consideration for early and late hyperkynetic arrhythmias. The most frequent, 4% of the patients, was in junctional tachycardia. Late arrhythmias can be atrial or ventricular. The incidence of ventricular arrhythmias in the literature range from 42 to 82%. In the Authors experience multiforme ventricular ectopy was present in 80% of the patients, 20 years after surgery. We suggest an exercise test and averaging ECG in all the patients. Thirty six percent of patients with supraventricular tachycardia were symptomatic. CONCLUSION: as the incidence of arrhythmias after correction of TOF is high, it is very important to periodically reassess these patients. Antiarrhythmic treatment is indicated in all symptomatic patients, especially in those with major arrhythmias (SVT, AF and VT).