Bulk RNA-Seq analysis to dissect the regulation of stigma position in tomato.

KEY MESSAGE: Transcriptomic analysis of tomato genotypes contrasting for stigma position suggests that stigma insertion occurred by the disruption of a process that finds a parallel in Arabidopsis gynoecium development. Domestication of cultivated tomato (Solanum lycopersicum L.) included the transition from allogamy to autogamy that occurred through the loss of self-incompatibilty and the retraction of the stigma within the antheridial cone. Although the inserted stigma is an established phenotype in modern tomatoes, an exserted stigma is still present in several landraces or vintage varieties. Moreover, exsertion of the stigma is a frequent response to high temperature stress and, being a cause of reduced fertility, a trait of increasing importance. Few QTLs for stigma position have been described and only one of the underlying genes identified. To gain insights on genes involved in stigma position in tomato, a bulk RNA sequencing (RNA-Seq) approach was adopted, using two groups of contrasting genotypes. Phenotypic analysis confirmed the extent and the stability of stigma position in the selected genotypes, whereas they were highly heterogeneous for other reproductive and productive traits. The RNA-Seq analysis yielded 801 differentially expressed genes (DEGs), 566 up-regulated and 235 down-regulated in the genotypes with exserted stigma. Validation by quantitative PCR indicated a high reliability of the RNA-Seq data. Up-regulated DEGs were enriched for genes involved in the cell wall metabolism, lipid transport, auxin response and flavonoid biosynthesis. Down-regulated DEGs were enriched for genes involved in translation. Validation of selected genes on pistil tissue of the 26 single genotypes revealed that differences between bulks could both be due to a general trend of the bulk or to the behaviour of single genotypes. Novel candidate genes potentially involved in the control of stigma position in tomato are discussed.

Pemetrexed plus platinum with or without pembrolizumab in patients with previously untreated metastatic nonsquamous NSCLC: protocol-specified final analysis from KEYNOTE-189.

BACKGROUND: In the phase III KEYNOTE-189 study (NCT02578680), pembrolizumab plus pemetrexed and platinum-based chemotherapy (pemetrexed-platinum) significantly improved overall survival (OS) and progression-free survival (PFS) in patients with previously untreated metastatic nonsquamous non-small-cell lung cancer (NSCLC) versus placebo plus pemetrexed-platinum. We report updated efficacy outcomes from the protocol-specified final analysis, including outcomes in patients who crossed over to pembrolizumab from pemetrexed-platinum and in patients who completed 35 cycles (∼2 years) of pembrolizumab. PATIENTS AND METHODS: Eligible patients were randomized 2 : 1 to receive pembrolizumab 200 mg (n = 410) or placebo (n = 206) every 3 weeks (for up to 35 cycles, ∼2 years) plus four cycles of pemetrexed (500 mg/m2) and investigators' choice of cisplatin (75 mg/m2) or carboplatin (area under the curve 5 mg·min/ml) every 3 weeks, followed by pemetrexed until progression. Patients assigned to placebo plus pemetrexed-platinum could cross over to pembrolizumab upon progression if eligibility criteria were met. The primary endpoints were OS and PFS. RESULTS: After a median follow-up of 31.0 months, pembrolizumab plus pemetrexed-platinum continued to improve OS [hazard ratio (HR), 0.56; 95% confidence interval (CI), 0.46-0.69] and PFS (HR, 0.49; 95% CI, 0.41-0.59) over placebo plus pemetrexed-platinum regardless of programmed death-ligand 1 expression. Objective response rate (ORR) (48.3% versus 19.9%) and time to second/subsequent tumor progression on next-line treatment (PFS2; HR, 0.50; 95% CI, 0.41-0.61) were improved in patients who received pembrolizumab plus pemetrexed-platinum. Eighty-four patients (40.8%) from the placebo plus pemetrexed-platinum group crossed over to pembrolizumab on-study. Grade 3-5 adverse events occurred in 72.1% of patients receiving pembrolizumab plus pemetrexed-platinum and 66.8% of patients receiving placebo plus pemetrexed-platinum. Fifty-six patients completed 35 cycles (∼2 years) of pembrolizumab; ORR was 85.7% and 53 (94.6%) were alive at data cut-off. CONCLUSIONS: Pembrolizumab plus pemetrexed-platinum continued to show improved efficacy outcomes compared with placebo plus pemetrexed-platinum, with manageable toxicity. These findings support first-line pembrolizumab plus pemetrexed-platinum in patients with previously untreated metastatic nonsquamous NSCLC.

Plasmonic Moon: A Fano-Like Approach for Squeezing the Magnetic Field in the Infrared.

Outstanding results have been achieved in the localization of optical electric fields via ultrasmall plasmonic cavities, paving the way to the subdiffractive confinement of local electromagnetic fields. However, due to the intrinsic constraints related to conventional architectures, no comparable squeezing factors have been managed yet for the magnetic counterpart of radiation, practically hindering the detection and manipulation of magneto-optical effects at the nanoscale. Here, we observe a strong magnetic field nanofocusing in the infrared, promoted by the induction of a coil-type Fano resonance. By triggering the coil current via a quadrupole-like plasmonic mode, we straightforwardly boost the enhancement of the infrared magnetic field and perform its efficient squeezing in localized nanovolumes.

Bleeding phenotype and correlation with factor XI (FXI) activity in congenital FXI deficiency: results of a retrospective study from a single centre.

Bleeding phenotype in factor XI (FXI)-deficient patients is variable, and not related to baseline FXI:Act. Aims of our study were to describe the characteristics and the management of surgery and deliveries in FXI-deficient patients, and to investigate the relationship between the haemorrhagic phenotype and the baseline FXI:Act. Ninety-five patients were diagnosed and followed in our centre for a median follow-up of 0.9 years (0.1-36.2); median FXI:Act of all patients: 38% (0.5-69%). Fifty-six patients (59%) experienced bleeding episodes not surgery-related. Prior to diagnosis, 64 patients underwent 132 surgeries, and after diagnosis, 23 patients underwent 36 surgeries. Globally 26 of 168 surgeries were prophylactically treated, whereas 142 of 168 were not. As regard as surgeries performed without prophylaxis, 30 bleeding events (21%) occurred in 21 patients. At diagnosis, the median FXI:Act of bleeding and non-bleeding patients was 28% and 37%, respectively, without statistically significant difference between the two groups (P = 0.26). As regard as surgeries performed under prophylactic treatment just 1 bleeding event occurred. Prior to diagnosis, 31 spontaneous deliveries (SD) and eight caesarian sections (CS) were performed without prophylaxis: 4 postpartum haemorrhages (10.5%) occurred (patients FXI:Act: 2%, 6%, 27%, 52.3% respectively). After diagnosis, four SD and five CS were performed with prophylaxis: no postpartum haemorrhages occurred. We confirm the wide bleeding phenotype variability in FXI-deficient patients, not related to the baseline FXI:Act levels. We highlight the importance of performing a correct diagnosis and follow-up, because a good management of prophylactic treatment, dramatically reduces the bleeding rate in case of surgery or deliveries.

Relationship between Prolactin Plasma Levels and White Matter Volume in Women with Multiple Sclerosis.

BACKGROUND: The role of prolactin (PRL) on tissue injury and repair mechanisms in multiple sclerosis (MS) remains unclear. The aim of this work was to investigate the relationship between PRL plasma levels and brain damage as measured by magnetic resonance imaging (MRI). METHODS: We employed a chemiluminescence immunoassay for measuring plasma levels of PRL. We used a 1.5 T scanner to acquire images and Jim 4.0 and SIENAX software to analyse them. RESULTS: We included 106 women with relapsing remitting (RR) MS and stable disease in the last two months. There was no difference in PRL plasma levels between patients with and without gadolinium enhancement on MRI. PRL plasma levels correlated with white matter volume (WMV) (rho = 0.284, p = 0.014) but not with grey matter volume (GMV). Moreover, PRL levels predicted changes in WMV (Beta: 984, p = 0.034). CONCLUSIONS: Our data of a positive association between PRL serum levels and WMV support the role of PRL in promoting myelin repair as documented in animal models of demyelination. The lack of an increase of PRL in the presence of gadolinium enhancement, contrasts with the view considering this hormone as an immune-stimulating and detrimental factor in the inflammatory process associated with MS.

Fano coil-type resonance for magnetic hot-spot generation.

The possibility to develop nanosystems with appreciable magnetic response at optical frequencies has been a matter of intense study in the past few years. This aim was strongly hindered by the saturation of the magnetic response of "natural" materials beyond the THz regime. Recently, in order to overcome such limitation, it has been considered to enhance the magnetic fields through the induction of displacement currents triggered by plasmonic resonances. Here we investigate a nanoassembly supporting the hybridization of an electric and magnetic plasmonic mode in Fano resonance conditions. Taking advantage of the enhancement properties owned by such interferential resonance, we have been able to generate an intense and localized magnetic hot-spot in the near-infrared spectral region.

Mood and coping in clinically isolated syndrome and multiple sclerosis.

OBJECTIVE: Few studies have examined behavioural changes in the early phase of multiple sclerosis (MS). The aim of the study is to investigate mood alterations and to explore coping strategies regarding patients with clinically isolated syndrome (CIS) and relapsing-remitting MS (RRMS). MATERIALS AND METHODS: The communication of diagnosis was made by one neurologist using a standardized approach. Depression, anxiety and coping questionnaires were filled in within 1 month from the diagnosis and at 3, 6, 12, 18 and 24 months subsequently. RESULTS: Thirty-nine patients were examined (11 CIS, 28 RRMS), also 39 healthy controls. At entry, patients showed a lower degree of hostile behaviour and a higher level of depression than the controls. At follow-up, a reduction in depression, anxiety and a better coping adjustment was observed. A higher reliance on 'Accepting responsibilities' coping score was seen in patients with higher levels of depression and anxiety. No significant differences were revealed by group comparisons between CIS and RRMS. CONCLUSIONS: This study highlights transient mood alterations and an improving of adaptive coping over a period of time in patients with CIS and RRMS. Similar emotional reactions and coping in clinical subgroups suggest that these factors are independent from the type of information provided during the communication of the diagnosis.

HLA-DQA1 and HLA-DQB1 genes in Tsachilas Indians from Ecuador: new insights in population analysis by Human Leukocyte Antigens.

Human Leucocyte Antigen (HLA) loci are widely known for their role in the generation of immune responses and are often considered to be effective in reconstructing human relationships. This is due to the high degree of polymorphism and the rarity of recombination observed at HLA loci. In this study, we have made an attempt to support the potential of HLA class II loci by analysing DQA1 and DQB1 in 52 Ecuadorians with ties to the Tsachilas community. Little is known about this populations either ethnologically or historically: they are considered retaining much of the ancient Chibchan culture in spite of the lack of significant genetic characterization. A total of 21 alleles were observed, with very low heterozygosity. The obtained data were then assessed for relationship reconstruction. The compiled database of 63 populations was segregated and resolved in clusters corresponding to the ethnogeographic distribution of the populations. This analysis of Central and Southern Amerindians allowed us to support a historical hypothesis related to the origin and migration of Ecuadorian people. Indeed, the relationships with neighbour human groups, especially Cayapas and Colombians, could shed light on the genetic similarity within ancient Chibchan culture that was dispersed by tribes coming up the Barbacoas. This indicates that if an appropriate analysis was to be carried out on a set of populations representative of different geographic locations, and that analysis was properly interpreted, then there would be a high possibility that HLA class II loci could infer accurate assessments, as revealed by uniparental markers.

Energy-level alignment in organic dye-sensitized TiO2 from GW calculations.

The electronic energy levels of some representative isolated and oxide-supported organic dyes, relevant for photovoltaic applications, are investigated using many-body perturbation theory within the GW approximation. We consider a set of all-organic dyes (denominated L0, L2, L3, and L4) featuring the same donor and anchor groups and differing for the linker moieties. We first calculate the energy levels of the isolated molecules, thus allowing us to address the effects of the different linker groups, and resulting in good agreement with photo-electron spectroscopic and electrochemical data. We then consider the L0 dye adsorbed on the (101) surface of anatase-TiO2. We find a density of occupied states in agreement with experimental photo-electron data. The HOMO-LUMO energy gap of the L0 dye is found to be reduced by ~1 eV upon adsorption. Our results validate the reliability of GW calculations for photovoltaic applications and point to their potential as a powerful tool for the screening and rational design of new components of electrochemical solar cells.

Bacterial ratchet motors.

Self-propelling bacteria are a nanotechnology dream. These unicellular organisms are not just capable of living and reproducing, but they can swim very efficiently, sense the environment, and look for food, all packaged in a body measuring a few microns. Before such perfect machines can be artificially assembled, researchers are beginning to explore new ways to harness bacteria as propelling units for microdevices. Proposed strategies require the careful task of aligning and binding bacterial cells on synthetic surfaces in order to have them work cooperatively. Here we show that asymmetric environments can produce a spontaneous and unidirectional rotation of nanofabricated objects immersed in an active bacterial bath. The propulsion mechanism is provided by the self-assembly of motile Escherichia coli cells along the rotor boundaries. Our results highlight the technological implications of active matter's ability to overcome the restrictions imposed by the second law of thermodynamics on equilibrium passive fluids.

Single-incision laparoscopic appendectomy is comparable to conventional laparoscopic and laparotomic appendectomy: our single center single surgeon experience.

Several studies have demonstrated the clinical and technical benefits of the laparoscopic surgery for complicated and uncomplicated appendicitis. Our retrospective study included 12 patient who underwent SILS appendectomy (SILS-A), 14 who received conventional laparoscopic surgery (VL-A), and 12 who received laparotomic appendectomy (OA); performed in all cases by the same surgeon (C.F.). The aim of this study was the comparison between this three different surgical techniques on same features: post operative leukocytosis, post operative pain, need abdominal drainage, esthetic viewpoint, incidence of complication, hospital stay. The results showed no significant differences between SILS-A and VLS-A, while an evident improvement shows versus O-A, even though not statistically significative. SILS was more effective in decreasing the risk of postoperative wound infection.

Genetic response to an environmental pathogenic agent: HLA-DQ and onchocerciasis in northwestern Ecuador.

The aim of this study is to explore human leukocyte antigen (HLA)-DQ variability in two populations (Cayapas Amerindians and Afro-Ecuadorians) who live near one another along the Cayapa River and who are exposed to the same environmental stresses, such as infection by Onchocerca volvulus. HLA-DQA1 and HLA-DQB1 of 149 unrelated individuals (74 Cayapas and 75 Afro-Ecuadorians) have been analyzed. HLA high-resolution molecular typing was performed by sequence-based typing, sequence-specific oligonucleotides hybridization and sequence-specific primer (SSP) amplification. The comparison between affected (cases) and unaffected people (controls) in both populations shows the key role of several HLA-DQA1 alleles in susceptibility and protection against onchocerciasis. In both populations, there is strong evidence related to the protective role of DQA1*0401 against onchocerciasis. Alleles HLA-DQA1*0102 and *0103 seem to represent risk factors in Afro-Ecuadorians, while HLA-DQA1*0301 is only a suggestive susceptibility allele in Cayapas. These findings represent new positive/negative associations with onchocerciasis in South America, whereas previous findings pertained only to African populations.

Electron-beam-driven collective-mode metamaterial light source.

We demonstrate experimentally that the energy from a highly localized free-electron-beam excitation can be converted via a planar plasmonic metamaterial to a low-divergence free-space light beam. This emission, which emanates from a collectively oscillating coupled metamolecule nanoantenna ensemble much larger in size than the initial excitation, is distinctly different from cathodoluminescence and bears some similarity with laser light. It offers a novel, flexible paradigm for the development of scalable, threshold-free light sources.

Histomorphologic alterations of human enamel after repeated applications of a bleaching agent.

The objective of the study was to analyse the histomorphology of enamel after repeated applications of a professional bleaching regimen. Enamel specimens were obtained from buccal surfaces of 20 extracted human incisors. Two specimens were obtained for each tooth. Half of each tooth was maintained in distilled water and served as control; the other part was treated with a 38 percent hydrogen peroxide professional bleaching agent. The treated specimens were divided in four groups: in group 1, the professional bleaching protocol suggested by the manufacturer was applied once; in group 2, the same protocol was repeated twice; in group 3, three times; in group 4, four times. Between bleaching applications and before SEM evaluation, enamel specimens were maintained in artificial saliva for 1 week. Enamel specimens for each group were submitted to a qualitative scanning electron microscopy (SEM) analysis (200X, 500X, 1000X, 3000X), comparing the treated specimens with the related control enamel. Results of the SEM analysis showed no relevant alteration on the enamel surfaces, when the bleaching protocol was applied once or twice. However, significant changes of enamel surface morphology were SEM observed in groups 3 and 4, suggesting a predominance of depressions when bleaching procedure was repeated three or four times. From the results of this in vitro study, it is possible to state that bleaching procedures should not be carried out indiscriminately. SEM analysis showed important alterations of the prismatic structure of the enamel when the bleaching protocol was applied three and four times.

Endodontic-orthodontic relationships: expression of no synthase in human dental pulp during orthodontic tooth movement.

Inflamed human pulp tissue presents an increase in the level of nitric oxide synthase (NOS). The aim of this study is to verify the presence of NOS in human pulp of teeth that are subject to orthodontic force. 20 healthy subjects, wearers of fixed braces on the upper arch, were selected. An open coil-spring in NiTi was applied on the upper premolar test tooth (TT); the controlateral control tooth (CCT) was subjected to orthodontic treatment but not to the further force of the open coil-spring; the antagonist control tooth (ACT) did not undergo any orthodontic treatment. Pulps were taken from test, contralateral control and antagonist control teeth immediately after the extractions which were done at 15 and 30 days from the start of application of the orthodontic force. The pulp tissue was analyzed through immunohistochemical and molecular biology examinations. The results showed tooth pulps subject to orthodontic treatment were very inflamed in the first 15 days with high levels of iNOS and low levels of eNOS; after 30 days a decrease of the inflammation and an increase of the pulp vascularization were observed together with a reduction of iNOS and an increase of eNOS respectively.

Long-term survival in cerebellar glioblastoma multiforme. Case report.

Primary cerebellar glioblastoma multiforme (GBM) is a rare tumor in adults that accounts for just 1% of all cases of GBM. Cerebellar GBMs are not yet completely understood also as far as the prognosis. We report a case of cerebellar GBM in a 27-year-old woman. Magnetic resonance imaging (MRI) showed a 3x3.6 cm-sized, ill-defined, heterogeneously enhancing mass in the left cerebellum. GBM was histologically confirmed following radical surgery. Postoperative radiotherapy with concomitant and adjuvant temozolomide chemotherapy was subsequently administrated. She has no evidence of recurrence and is in good clinical conditions up-to date, three years after surgery.

Overexpression of interleukin-6 and -8, cell growth inhibition and morphological changes in 2-hydroxyethyl methacrylate-treated human dental pulp mesenchymal stem cells.

AIM: To evaluate morphological features, cell growth and interleukin-6 (IL-6) and interleukin-8 (IL-8) secretion in expanded ex vivo human dental pulp mesenchymal stem cells (DP-MSCs) after exposure to 2-hydroxyethyl methacrylate (HEMA). METHODOLOGY: Dental pulp mesenchymal stem cells were derived from the dental pulps of 10 young donors. After in vitro isolation, DP-MSCs were treated with 3 and 5 mmol L(-1) HEMA, and after 24, 48 and 72 h of incubation, their morphological features, cell growth, IL-6 and IL-8 secretion were analysed. Differences in the cell growth and in the interleukin secretion were analysed for statistical significance with two-way anova tests and the Holm-Sidak method for multiple comparisons. RESULTS: Dental pulp mesenchymal stem cells revealed a decrease in cell growth with both treatments (P < 0.05), more evident at 5 mmol L(-1) . Microscopic analysis displayed extensive cytotoxic effects in treated cells, which lost their fibroblastoid features and became retracted, even roundish, with a large number of granules. An up-regulation of IL-6 and IL-8 in treated cells cytokines was evident (P < 0.05). CONCLUSIONS: 2-Hydroxyethyl methacrylate exhibited cytotoxicity, inhibited cell growth and induced morphological changes in cultured DP-MSCs. Moreover, in treated samples, an up-regulation of soluble mediators of inflammation such as IL-6 and IL-8 cytokines was found. The direct application of HEMA potentially induces an inflammation process that could be the starting point for toxic response and cell damage in DP-MSCs.

Conservative restorations of endodontically compromised anterior teeth in paediatric patients: physical and mechanical considerations.

AIM: Objective of this in vitro study was to evaluate the influence of fiber post placement on fracture resistance of pulpless anterior teeth restored with standardised Class III and Class IV resin composite fillings. MATERIALS AND METHODS: One hundred and five human maxillary central incisors were selected and randomly divided into 7 (n=15) experimental groups (endodontic therapy/ endodontic therapy and one Class III resin composite filling/ endodontic therapy and one Class IV resin composite filling/ endodontic therapy and two Class III resin composite fillings/ endodontic therapy, fiber post and one Class III resin composite filling/ endodontic therapy, fiber post and one Class IV resin composite filling/ endodontic therapy, fiber post and two Class III resin composite fillings). Specimens underwent fracture strength test. Means (N) were calculated and data were analysed using 1-way ANOVA and Tukey multiple comparisons tests (p=0.05). RESULTS: Concerning teeth with two Class III, fiber post placement significantly increased fracture strength values from 603.59 to 864.24 N. Specimens restored with one Class III (795.21 N without post, 936.68 N with post) showed higher fracture strength values if compared with specimens with two Class III, with significant differences just concerning specimens without a fiber post. Fracture strength was not significantly influenced by fiber post placement in Class IV groups (720.71 N without post, 799.69 N with post). CONCLUSION: Data suggest that fiber post placement may significantly improve anterior teeth fracture strength when at least two Class III composite fillings are associated to the endodontic treatment. No significant effect of fiber post placement could be recorded when just one Class III or one Class IV composite filling were present.

Liver resection for hemoperitoneum caused by spontaneous rupture of unrecognized hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is an increasingly common form of cancer. Although its spontaneous rupture is rare in Western countries, it constitutes a surgical emergency and is associated with high mortality. There is a lack of consensus as to the best approach and what parameters to use in choosing it. The three main approaches are conservative, endovascular and resection - the treatment of choice for acute abdominal bleeding. We report a case of hemoperitoneum following the spontaneous rupture of an unrecognized HCV-related HCC in a patient with no history of liver disease. The patient was successfully treated by emergency surgery, with resection of two segments of the left liver.

Multi-scheme approach for efficient surface plasmon polariton generation in metallic conical tips on AFM-based cantilevers.

We report on the possibility of realizing adiabatic surface plasmon polaritons compression on metallic conical tips built-in on AFM cantilevers by means of different approaches. The problem is faced considering the role of the source, when linear and radial polarizations are assumed, associated to different fabrication schemes. Nano-patterned devices properly combined with metallic conical tips can affect the adiabatic characteristic of the surface electric field. The results are analyzed in terms of tradeoff between fabrication difficulties and device performances. Suggestions on the best possible scheme are provided.