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Aim: To delineate clinical correlates of COVID-19 infection severity in hospitalized patients with malignancy. Methods: The authors conducted a retrospective review of all hospitalized patients with a hematologic and/or solid tumor malignancy presenting to the authors' institution between 1 March 2020 and 5 January 2021, with a laboratory confirmed diagnosis of COVID-19. Univariate and multivariate logistic regression analyses were used to determine associations between specific severity outcomes and clinical characteristics. Results: Among 2771 hospitalized patients with COVID-19, 246 (8.88%) met inclusion criteria. Patients who were actively receiving treatment had an increased rate of death following admission (odds ratio [OR]: 2.7). After adjusting for significant covariates, the odds ratio increased to 4.4. Patients with cancer involvement of the lungs had a trend toward increased odds of death after adjusting for covariates (OR: 2.3). Conclusions: Among COVID-19 positive hospitalized cancer patients, systemic anti-cancer therapy was associated with significantly increased odds of mortality.

Volcanic activity at Mt. Etna in the last decade has mostly been manifested by sequences of short paroxysmal episodes characterised by powerful lava fountains and high eruption columns. On the 23 February 2013, an exceptionally intense episode occurred at the New South-East Crater, producing a fountain >800 m high (among the highest ever recorded at Etna) and a ~9 km eruption column that dispersed ash >400 km from the vent. Textural and petrographic analyses of lapilli revealed that magma erupted during the high-intensity phase is characterised by low microlite contents (

Background Grapevine reproductive development has direct implications on yield. It also impacts on berry and wine quality by affecting traits like seedlessness, berry and bunch size, cluster compactness and berry skin to pulp ratio. Seasonal fluctuations in yield, fruit composition and wine attributes, which are largely driven by climatic factors, are major challenges for worldwide table grape and wine industry. Accordingly, a better understanding of reproductive processes such as gamete development, fertilization, seed and fruit set is of paramount relevance for managing yield and quality. With the aim of providing new insights into this field, we searched for clones with contrasting seed content in two germplasm collections. Results We identified eight variant pairs that seemingly differ only in seed-related characteristics while showing identical genotype when tested with the GrapeReSeq_Illumina_20K_SNP_chip and several microsatellites. We performed multi-year observations on seed and fruit set deriving from different pollination treatments, with special emphasis on the pair composed by Sangiovese and its seedless variant locally named Corinto Nero. The pollen of Corinto Nero failed to germinate in vitro and gave poor berry set when used to pollinate other varieties. Most berries from both open- and cross-pollinated Corinto Nero inflorescences did not contain seeds. The genetic analysis of seedlings derived from occasional Corinto Nero normal seeds revealed that the few Corinto Nero functional gametes are mostly unreduced. Moreover, three genotypes, including Sangiovese and Corinto Nero, were unexpectedly found to develop fruits without pollen contribution and occasionally showed normal-like seeds. Five missense single nucleotide polymorphisms were identified between Corinto Nero and Sangiovese from transcriptomic data. Conclusions Our observations allowed us to attribute a seedlessness type to some variants for which it was not documented in the literature. Interestingly, the VvAGL11 mutation responsible for Sultanina stenospermocarpy was also discovered in a seedless mutant of Gouais Blanc. We suggest that Corinto Nero parthenocarpy is driven by pollen and/or embryo sac defects, and both events likely arise from meiotic anomalies. The single nucleotide polymorphisms identified between Sangiovese and Corinto Nero are suitable for testing as traceability markers for propagated material and as functional candidates for the seedless phenotype.

Background This retrospective single center study aimed to describe circulating tumor DNA (ctDNA) comprehensive genomic profiling (CGP) utilization in a community practice for patients with advanced solid tumors. Methods All patients were included who were seen at the Scripps Hillcrest Oncology Clinic (San Diego, CA, USA) between September 2016 to March 2018 who had ctDNA assay testing performed. In this cohort, all ctDNA testing was performed to aid therapeutic decision making with wide variety in both the type of advanced solid tumor, as well as the line of therapy. Results Of the assays performed in the 41 patients included in this review, 42% of therapeutic actions following ctDNA assay results were influenced by the ctDNA result, including initiation of the corresponding Federal Drug Administration (FDA) approved therapy, placement on clinical trial, and initiation of off label-targeted options. In addition, CGP results guided clinicians away from futile or harmful treatments, such as EGFR inhibition in colorectal cancer patients with discovered KRAS mutations. No additional prognostic or therapeutic information was gathered in one quarter of patients for which ctDNA was drawn. Furthermore, discovered genomic alterations by ctDNA testing did not influence therapeutic action in 58% of cases. Conclusions These results highlight the conundrum that having additional information regarding an individual’s tumor biology does not yet translate into meaningful targeted therapy in the majority of cases. Further studies are needed regarding ctDNA utilization to help guide community oncologists who will continue to face the choice between targeted therapy, immunotherapy, and cytotoxic chemotherapy as science advances.

International audience; AbstractIntroduction : Celiac disease is estimated to affect between 1 in 100 and 1 in 300 Caucasian subjects. Many quantitative studies have explored the issues encountered by these subjects; however, few studies have addressed the subjective and experiential dimensions of celiac disease.Objective : The objective of this qualitative exploratory study is to expand current knowledge concerning the subjective experience of celiac disease and of a gluten-free diet in subjects diagnosed with celiac disease in adulthood.Method : Data was collected through non-directive interviews with 14 subjects aged between 28 and 53 (M = 41.7, SD = 7.48).Results : An interpretive phenomenological analysis identified various themes related to the experience of illness: (1) history of the disease, (2) symptoms, (3) nutrition, (4) perception of medical experiences, (5) relationships with family and friends, (6) psychological implications, and (7) consequences for daily living.Conclusion : The results show the importance of developing a “mythology” (concerning the disease's origin) to enable the inclusion of the condition in the individual's on-going personal story.

We present a multidisciplinary study of a fossiliferous site located in the Vulsini Volcanic District, on the western side of the Tiber River Valley north of Rome, highlighting the peculiar geologic factors that contributed to the origin and preservation of an outstanding archaeological record testifying of the early human frequentation in this region. Mighty explosive eruptions since at least 500 ka affected the investigated area eventually culminating in the formation of the huge Bolsena caldera. Tectonic deformation accompanying volcanic activity caused large fault displacements, shaping the ground surface and contributing to route the path, and possibly to trigger, the catastrophic emplacement of volcaniclastic flows. A sedimentary trap originated by fault scarp cutting through a streambed was likely the cause for the large accumulation of bones and stone artifacts ripped up and carried by a volcaniclastic flow at 322 ka. The analysis of the fossil assemblage reveals both gnawing traces by carnivores and cut-marks from the percussion tools employed by humans to butch the carcasses. However, the occurrence of retouched and unretouched blanks within the lithic assemblage also testifies for provenance from a wider area of human activity, which included hunting and scavenging, probably at a nearby butchering site.

BackgroundReproductive success in seed plants depends on a healthy fruit and seed set. Normal seed development in the angiosperms requires the production of functional female gametophytes. This is particularly evident in seedless cultivars where defects during megagametophyte’s developmental processes have been observed through cytohistological analysis. Several protocols for embryo sac histological analyses in grapevine are reported in literature, mainly based on resin- or paraffin-embedding approaches. However their description is not always fully exhaustive and sometimes they consist of long and laborious steps. The use of different stains is also documented, some of them, such as hematoxylin, requiring long oxidation periods of the dye-solution before using it (from 2 to 6 months) and/or with a differentiation step not easy to handle. Paraffin-embedding associated to examination with light microscope is the simplest methodology, and with less requirements in terms of expertise and costs, achieving a satisfactory resolution for basic histological observations. Safranin O and fast green FCF is an easy staining combination that has been applied in embryological studies of several plant species.ResultsHere we describe in detail a paraffin-embedding method for the examination of grapevine ovules at different phenological stages. The histological sample preparation process takes 1 day and a half. Sections of 5 µm thickness can be obtained and good contrast is achieved with the safranin O and fast green FCF staining combination. The method allows the observation of megasporogenesis and megagametogenesis events in the different phenological stages examined.ConclusionsThe histological sample preparation process proposed here can be used as a routine procedure to obtain embedded ovaries or microscope slides that would require further steps for examination. We suggest the tested staining combination as a simple and viable technique for basic screenings about the presence in grapevine of a normally and fully developed ovule with embryo sac cells, which is therefore potentially functional.

Purpose: Prolonged weightlessness exposure generates cardiovascular deconditioning, with potential implications on ECG circadian rhythms. Head-down (- 6°) tilt (HDT) bed rest is a ground-based analogue model for simulating the effects of reduced motor activity and fluids redistribution occurring during spaceflight. Our aim was to evaluate the impact of 60-day HDT on the circadianity of RR and ventricular repolarization (QTend) intervals extracted from 24-h Holter ECG recordings, scheduled 9 days before HDT (BDC-9), the 5th (HDT5), 21st (HDT21) and 58th (HDT58) day of HDT, the 1st (R + 0) and 8th (R + 7) day after HDT. Also, the effectiveness of a nutritional countermeasure (CM) in mitigating the HDT-related changes was tested. Methods: RR and QTend circadian rhythms were evaluated by Cosinor analysis, resulting in maximum and minimum values, MESOR (a rhythm-adjusted mean), oscillation amplitude (OA, half variation within a night-day cycle), and acrophase (?, the time at which the fitting sinusoid's amplitude is maximal) values. Results: RR and QTend MESOR increased at HDT5, and the OA was reduced along the HDT period, mainly due to the increase of the minima. At R + 0, QTend OA increased, particularly in the control group. The ? slightly anticipated during HDT and was delayed at R + 0. Conclusion: 60-Day HDT affects the characteristics of cardiac circadian rhythm by altering the physiological daily cycle of RR and QTend intervals. Scheduled day-night cycle and feeding time were maintained during the experiment, thus inferring the role of changes in the gravitational stimulus to determine these variations. The applied nutritional countermeasure did not show effectiveness in preventing such changes.

Introduction Prolonged bed rest (BR) is an unnatural state, often related to hospitalization, chronic diseases and ageing, inducing reduced functional capacity in multiple body systems, possibly leading to cardiovascular deconditioning. We hypothesized that measuring this decline over time could represent the first step for the formulation of appropriate countermeasures or rehabilitation programs while in the hospital. Accordingly, our aim was to assess the effects of 10-day horizontal BR on cardiac electrical activity. Methods Ten healthy male volunteers (23±5 years) were enrolled in an hospital, after ethical approval and signed consent, to participate to a 10-day strict horizontal BR campaign, preceded and followed by 2 days in the facility, respectively as acclimatization and recovery. The 12-leads 24-hours Holter ECG (1000 Hz, H12+, Mortara Instrument Inc.) was acquired 1 day before BR (PRE), the 5th (BR5) and 10th day (BR10) of bedridden immobilization. From each recording, beat-to-beat RR and QTend interval series, as well as T wave amplitude (Tamp) and upslope (Tslope) were computed. Statistical analysis was applied to test changes induced by BR (ANOVA with Tukey test, p Results Daily RR and QTend duration increased during BR, with peak changes at BR5 compared to PRE (+13.3% and +3% respectively), and were still prolonged at BR10 (+12.6% and +2.6%). During the night, while RR increased (BR5:+5.3%; BR10:+1.3%), QTend was found progressively shortened (BR5: −1.6%; BR10: −2.9%). Also, day and night Tamp (BR10: −19.5%) and Tslope (BR10 day: −17.1%; night: −7.8%) were found progressively reduced with the duration of BR. Conclusion During BR, cardiac electrical activity is affected by 10-days bedridden immobilization. Noticeably, a mismatch in RR-QTend relation was visible at night, where vagal autonomic system activity is prevailing. Funding Acknowledgement Type of funding source: Other. Main funding source(s): Agenzia Spaziale Italiana (ASI)

Aims Prolonged immobilization generates cardiac deconditioning, a risk factor for cardiovascular disease, and efficient countermeasures (CM) are needed to prevent it. Our aim was to assess by Cine-MRI the effects of long-term strict head-down (−6 degrees) bed-rest (BR) deconditioning, and the effectiveness of high-intensity jump training CM, on left ventricular (LV) function and mass. Methods 23 male participants (29±6 years, 181±6 cm, 77±7 kg) were enrolled. The experiment was conducted at: envihab (Koln, DLR, Germany) as part of the European Space Agency BR studies. Volunteers were randomly allocated to the jump training group (JUMP, n=12) or the control group (CTRL, n=11). A typical training session consisted of 4x10 countermovement jumps and 2x10 hops in a sledge jump system, with 5–6 sessions per week. Steady-state free precession cine-MRI images were obtained (25 frames/cardiac cycle, 8mm thickness, no gap, no overlap) as stack of short-axis images covering the whole LV from base to apex, before (PRE) and after 58-days (HDT58) of BR. Endocardial and epicardial semi-automated contouring was performed using custom software. Results In CTRL group, at HDT58 a reduction in LV mass (9%), end-diastolic (21%), end-systolic (8%) and stroke volume (23%) were observed, while ejection fraction did not change. In JUMP group, the reduction in LV end-diastolic volume was only by 9%, followed by a decrease in end-systolic (10%) and stroke volume (10%), with a preservation of LV mass. In both groups, ejection fraction did not change. Left ventricular changes after 58d BR EDV (ml) ESV (ml) SV (ml) EF (%) Mass (g) CTRL PRE 171 (163; 191) 60 (58; 65) 113 (102; 124) 64 (63; 67) 133 (121; 160) HDT58 143 (131; 149)* 56 (50; 58)* 86 (83; 92)* 62 (60; 63)* 127 (106; 138)* JUMP PRE 156 (140; 204) 70 (61; 84) 91 (78; 117) 58 (56; 60) 118 (104; 134) HDT58 141 (135; 175)* 63 (54; 69)* 90 (78; 92)* 58 (55; 60) 113 (105; 128) Results expressed as median (25th; 75th percentiles). *p Conclusions Cardiac adaptation to deconditioning due to immobilization induced by BR resulted in a reduction of cardiac volumes and function, together with a decrease in LV mass. Interestingly, the applied JUMP countermeasure appeared able to partially reverse these effects, in particular by reducing the decrease in end-diastolic volume and preserving LV mass. This information could be useful for better understanding physiologic changes in patients undergoing long periods of immobilization, as well as to apply the studied countermeasure during space flight to reduce cardiac deconditioning. Acknowledgement/Funding Italian Space Agency (contract 2018-7-U.0),CNES/DAR 48ehz747.01950965,BELSPO, via the European Space Agency PRODEX program (PEA 4000110826)

In the present paper, we analyze the regional interplay between glacio-eustasy and volcano-tectonic uplift on the Tyrrhenian Sea margin of central Italy. We reconstruct a succession of fluvial terraces in the Tiber River Valley east of the Vulsini Volcanic District and we provide geochronologic constraints allowing for correlation with the sea-level highstands of the Marine Isotope Stage timescale. Results of this study show that glacio-eustatic forcing affected the hydrographic network of the Tiber River, as far as 70 km inland with respect to the present Tyrrhenian coast, consistent with a regional uplift on the order of several tens of meters that affected this region over the last 250 ky. Using six new 40Ar/39Ar dates, we demonstrate the synchronicity between sea-level rise during glacial termination IV and the deposition of a fining-upward sedimentary succession of the Paleo-Tiber River in this area. A detailed reconstruction of the chronostratigraphic setting enabled us to assess local uplift and fault displacement due to volcano-tectonic processes associated with the activities of Bolsena-Orvieto and Latera volcanoes since 350 ka, and develop an uplift curve for the different sectors of the investigated area. Moreover, we estimate sedimentation rate during post-glacial sea-level rise. Strong differential uplift, with rates on the order of 0.6–1.2 mm/yr corresponded with the onset of major eruptive phases, whereas a homogeneous regional uplift of 0.24 mm/yr during the last 125 ky followed this climactic phase. These uplift rates are significantly smaller than sedimentation rate during the glacial termination, consistent with the observed independence of the glacio-eustatic signal.

The optimal balance between sugar and acidity is an essential criterion to elaborate equilibrated and stable wines. The aim of this study was to locate quantitative trait loci (QTLs) for these traits using an F1 population derived from Monastrell and Syrah wine cultivars. Several parameters related to acidity were evaluated during six consecutive years by measuring total soluble solids, total acidity, malic acid, and tartaric acid. Three genetic maps were developed using 104 SSR (simple sequence repeat) and 146 SNP (single-nucleotide polymorphism) markers. The consensus map covered 1174 cM with 238 markers assembled in 19 linkage groups (LGs). Significant QTLs at the genome-wide level were detected, and, although they exhibited a large degree of instability from year to year, QTLs for the ratio of soluble solids to acidity (LG2) and malic acid (LG8) and the ratio of tartaric to malic acid (LG8) were stable in at least 2 years. Several annotated genes involved in sugar and acidity pathways co-located with the confidence intervals of these QTLs and are proposed as putative candidate genes for future studies of these traits.

Highlight Novel candidate genes for the fine regulation of flavonol content in ripe berries are identified through integration of transcriptional profiling and metabolic QTL analyses of a segregating grapevine progeny., Flavonols are a ubiquitous class of flavonoids that accumulate preferentially in flowers and mature berries. Besides their photo-protective function, they play a fundamental role during winemaking, stabilizing the colour by co-pigmentation with anthocyanins and contributing to organoleptic characteristics. Although the general flavonol pathway has been genetically and biochemically elucidated, the genetic control of flavonol content and composition at harvest is still not clear. To this purpose, the grapes of 170 segregating F1 individuals from a ‘Syrah’×’Pinot Noir’ population were evaluated at the mature stage for the content of six flavonol aglycons in four seasons. Metabolic data in combination with genetic data enabled the identification of 16 mQTLs (metabolic quantitative trait loci). For the first time, major genetic control by the linkage group 2 (LG 2)/MYBA region on flavonol variation, in particular of tri-hydroxylated flavonols, is demonstrated. Moreover, seven regions specifically associated with the fine control of flavonol biosynthesis are identified. Gene expression profiling of two groups of individuals significantly divergent for their skin flavonol content identified a large set of differentially modulated transcripts. Among these, the transcripts coding for MYB and bZIP transcription factors, methyltranferases, and glucosyltranferases specific for flavonols, proteins, and factors belonging to the UV-B signalling pathway and co-localizing with the QTL regions are proposed as candidate genes for the fine regulation of flavonol content and composition in mature grapes.

Aims: Prolonged immobilization generates cardiac deconditioning, a cardiovascular disease risk factor, and efficient countermeasures (CM) are needed to prevent it. We aimed to assess by Phase-Contrast (PC) MRI the effects of long-term strict head-down (-6 degrees) bed-rest (BR) deconditioning and the effectiveness of high-intensity jump training CM on aortic and mitral flow. Methods: 23 males (29±6 years, 181±6 cm, 77±7 kg) were enrolled. The experiment was conducted at Envihab research facility as part of the European Space Agency BR studies. Participants were randomly allocated to the jump training group (JUMP, n=12) or to the control group (CTRL, n=11). A typical training session consisted of 4x10 countermovement jumps and 2x10 hops in a horizontal sledge jump system, with 5–6 sessions/week. PC-MRI images (3T Biograph mMR) with interleaved 3-directional velocity encoding (VENC: x and y: 80 cm/s; z: 150 cm/s) were obtained (spatial resolution 1.4x1.4 mm2) at the level of the aortic root, and of the mitral plane, before (PRE) and after 58-days (HDT58) of BR. The resulting planar magnitude data and 3-directional velocity images were semi-automatically analysed with previously validated custom software to compute the following parameters: cardiac output (CO), stroke volume (SV), flow rate (Qpeak), systolic duration (Sys) and heart beat duration (RR), rapid filling (Efill) and inflow rate (Epeak). Results: In CTRL, compared to baseline values, at HDT58 a significant (p

Con autorización de la revista para autores CSIC, [EN] The BRAIN (Botanical Records of Archaeobotany Italian Network) database and network was developed by the cooperation of archaeobotanists working on Italian archaeological sites. Examples of recent research including pollen or other plant remains in analytical and synthetic papers are reported as an exemplar reference list. This paper retraces the main steps of the creation of BRAIN, from the scientific need for the first research cooperation to the website which has a free online access since 2015.

The external profile of seeds is a good phenotypic descriptor, indicative of the origin and diffusion of grape vines (Vitis vinifera L.) over the centuries. Archaeological seeds conserved in an anaerobic environment after discovery were compared with modern seeds, belonging to eleven autochthonous cultivars, sampled in Tuscan vineyards. To study seed profiles we used geometric morphometric analysis (elliptical Fourier analysis) combined with multivariate statistical methods. When the values obtained were entered in a database, they produced a cladogram based on Euclidean distances between varieties of archaeological and modern vines, showing that seeds belonging to the same group had phenotypic affinities.

Since the 1990s, oral chemotherapy has been gaining ground as cancer treatment. This therapy seems to have few toxic effects and offers patients good quality of life. However, in addition to the fears the therapy might generate in patients, oral treatment raises a new issue, which, until now, has been marginal in this field: therapeutic observance or adherence. We investigated the research into adherence to oral chemotherapy among cancer patients published between 1990 and July 2013. Studies showed considerable diversity in terms of both the definition and measurement of adherence. As well, adherence to antineoplastic therapy is affected by the patient's understanding of the treatment and ability to remember information provided by the physician, treatment length and psychological distress. Our review of the few studies on adherence to anticancer drug treatment raises some questions that could be pursued in future research. In light of our findings, patients should receive 'therapy education' to help them and their support groups better understand the disease and its treatment and to achieve optimal health management and improved treatment effectiveness.

The incidence of detecting focal chronic recurrent multifocal osteomyelitis (CRMO) lesions on dual-energy x-ray absorptiometry (DXA), and the effect of these lesions on DXA bone mineral density (BMD), bone mineral content (BMC), and their associated Z-scores were retrospectively reviewed. Materials and Methods. The study included 22 patients (14 females, 8 males; median age of 13 years) with CRMO and in whom a total body less head (TBLH) and lumbar spine DXA scan had been obtained. Whole-body bone scintigraphy and MRI were used as the reference standards. Sites involved with CRMO were subsequently detected and DXA measurements were re-measured after removing the sclerotic lesions from the analysis. Results. In total, sclerotic CRMO lesions were detected in 15 of the 22 patients (68%) by DXA, although the number of lesions detected (on a per-lesion analysis) was much less (i.e. 29 of 129 lesions; 19.4%) when compared to MRI and/or bone scintigraphy. Larger lesions had a greater impact on the derived BMD/BMC measurements, and changed the diagnosis in one patient from having normal to abnormal DXA results based on the final Z-score. Discussion. CRMO lesions detected on DXA examinations should be regarded as a potential source of error. Careful inspection and re-quantification of the BMD, BMC and associated Z-score after applying an appropriate correction should be considered in patients with large CRMO lesions identified on DXA examinations.

UNLABELLED: Pertuzumab is a HER2 dimerization inhibitor that binds to an epitope unique from that of trastuzumab. Our objective was to determine whether SPECT with (111)In-diethylenetriaminepentaacetic acid-pertuzumab ((111)In-DTPA-pertuzumab) could sensitively detect an early molecular response to trastuzumab manifested by HER2 downregulation and a later tumor response revealed by a decreased number of HER2-positive viable tumor cells. METHODS: Changes in HER2 density in SKBr-3 and MDA-MB-361 BC cells exposed to trastuzumab (14 microg/mL) in vitro were measured by saturation binding assays using (111)In-DTPA-pertuzumab and by confocal immunofluorescence microscopy and flow cytometry with fluorescein isothiocyanate-labeled HER2/neu antibodies. Imaging of HER2 downregulation was studied in vivo in athymic mice with subcutaneous MDA-MB-361 tumors treated for 3 d with trastuzumab (4 mg/kg) or nonspecific human IgG (hIgG) or phosphate-buffered saline (PBS). Imaging of tumor response to trastuzumab was studied in mice bearing subcutaneous MDA-MB-361 xenografts treated with trastuzumab (4 mg/kg), followed by weekly doses of nonspecific hIgG or rituximab or PBS (2 mg/kg). Mice were imaged on a micro-SPECT/CT system at 72 h after injection of (111)In-DTPA-pertuzumab. Tumor and normal-tissue biodistribution was determined. RESULTS: (111)In-DTPA-pertuzumab saturation binding to SKBr-3 and MDA-MB-361 cells was significantly decreased at 72 h after exposure in vitro to trastuzumab (14 microg/mL), compared with untreated controls (62% +/- 2%, P < 0.0001; 32% +/- 9%, P < 0.0002, respectively). After 3 d of trastuzumab, in vivo tumor uptake of (111)In-DTPA-pertuzumab decreased 2-fold in trastuzumab- versus PBS-treated mice (13.5 +/- 2.6 percentage injected dose per gram [%ID/g] vs. 28.5 +/- 9.1 %ID/g, respectively; P < 0.05). There was also a 2-fold decreased tumor uptake in trastuzumab- versus PBS-treated mice by image volume-of-interest analysis (P = 0.05), suggesting trastuzumab-mediated HER2 downregulation. After 3 wk of trastuzumab, tumor uptake of (111)In-DTPA-pertuzumab decreased 4.5-fold, compared with PBS-treated mice (7.6 +/- 0.4 vs. 34.6 +/- 9.9 %ID/g, respectively; P < 0.001); this decrease was associated with an almost-completed eradication of HER2-positive tumor cells determined immunohistochemically. CONCLUSION: (111)In-DTPA-pertuzumab sensitively imaged HER2 downregulation after 3 d of treatment with trastuzumab and detected a reduction in viable HER2-positive tumor cells after 3 wk of therapy in MDA-MB-361 human breast cancer xenografts.

Tandem pleckstrin homology domain proteins (TAPPs) are recruited to the plasma membrane via binding to phosphoinositides produced by phosphoinositide 3-kinases (PI3Ks). Whereas PI3Ks are critical for B-cell activation, the functions of TAPP proteins in B cells are unknown. We have identified 40 potential interaction partners of TAPP2 in B cells, including proteins involved in cytoskeletal rearrangement, signal transduction and endocytic trafficking. The association of TAPP2 with the cytoskeletal proteins utrophin and syntrophin was confirmed by Western blotting. We found that TAPP2, syntrophin, and utrophin are coexpressed in normal human B cells and B-chronic lymphocytic leukemia (B-CLL) cells. TAPP2 and syntrophin expression in B-CLL was variable from patient to patient, with significantly higher expression in the more aggressive disease subset identified by zeta-chain–associated protein kinase of 70 kDa (ZAP70) expression and unmutated immunoglobulin heavy chain (IgH) genes. We examined whether TAPP can regulate cell adhesion, a known function of utrophin/syntrophin in other cell types. Expression of membrane-targeted TAPP2 enhanced B-cell adhesion to fibronectin and laminin, whereas PH domain–mutant TAPP2 inhibited adhesion. siRNA knockdown of TAPP2 or utrophin, or treatment with PI3K inhibitors, significantly inhibited adhesion. These findings identify TAPP2 as a novel link between PI3K signaling and the cytoskeleton with potential relevance for leukemia progression.

Signaling via phosphoinositide 3-kinases (PI3Ks) has emerged as a central component of lymphocyte activation via immunoreceptors, costimulatory receptors, cytokine receptors, and chemokine receptors. The discovery of phosphoinositide-binding pleckstrin homology (PH) domains has substantially increased understanding of how PI3Ks activate cellular responses. Accumulating evidence indicates that PH-domain containing adapter molecules provide important links between PI3K and lymphocyte function. Here, we review data on PI3K-regulated adapter proteins of the Grb-associated binder (GAB), Src kinase-associated phosphoprotein (SKAP), and B-lymphocyte adapter molecule of 32 kDa (Bam32)/ dual-adapter for phosphotyrosine and 3-phosphoinositides (DAPP)/TAPP families, with a focus on the latter group. Current data support the model that recruitment of these adapters to the plasma membrane of activated lymphocytes is driven by the phosphoinositides phosphatidylinositol-3,4,5-tris-phosphate and phosphatidylinositol-3,4-bisphosphate, generated through the action of PI3Ks and under the regulatory control of lipid phosphatases Src homology 2 domain-containing inositol phosphatase (SHIP), phosphatase and tensin homolog, and inositol polyphosphate 4-phosphatase. At the plasma membrane, these adapters serve to assemble distinct protein complexes. Bam32/DAPP1 and SKAPs function to promote activation of monomeric guanosine triphosphatases, including Rac and Rap, and promote integrin activation, lymphocyte adhesion to matrix proteins, and cell:cell interactions between B and T lymphocytes. GABs can provide feedforward amplification or feedback inhibition of PI3K signaling. Current work is further defining the molecular interactions driven by these molecules and identifying the functions of TAPP adapters, which also appear to be involved in lymphocyte adhesion and are specific effectors downstream of the SHIP product phosphatidylinositol-3,4-bisphosphate.

Conservation of grapevine biodiversity in the Caucasus and Northern Black Sea Region is particularly urgent because of: 1) the large number of traditional local varieties out of cultivation; 2) the relevance of these resources for the development of European modern cultivars; 3) the financial difficulties in the countries; 4) the occurrence of Vitis vinifera ssp. silvestris throughout the region; 5) wine production as a major potential source of income for the local population in the region. In 2004-2005, significant progress has been made within a collaborative project aimed at strengthening the capacity of the countries of the region (Armenia, Azerbaijan, Georgia, Moldova, Russia, and Ukraine) to ensure the long-term maintenance of Vitis genetic resources, including the cultivated traditional varieties and the wild resources. The activities include identifying, collecting, characterizing, and conserving the diversity of grapevine genetic resources as a basis to improve local viticulture and winemaking industry.

UNLABELLED: Our goals in this study were to determine whether (111)In-trastuzumab coupled to peptides harboring nuclear localizing sequences (NLSs) could kill trastuzumab-resistant breast cancer cell lines through the emission of Auger electrons and whether the combination of radiosensitization with methotrexate (MTX) would augment the cytotoxicity of this radiopharmaceutical. METHODS: Trastuzumab was derivatized with sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate for reaction with NLS peptides and then conjugated with diethylenetriaminepentaacetic acid for labeling with (111)In. HER2 expression was determined by Western blot and by radioligand binding assay using (111)In-trastuzumab in a panel of breast cancer cell lines, including SK-BR-3, MDA-MB-231 and its HER2-transfected subclone (231-H2N), and 2 trastuzumab-resistant variants (TrR1 and TrR2). Nuclear importation of (111)In-NLS-trastuzumab and (111)In-trastuzumab in breast cancer cells was measured by subcellular fractionation, and the clonogenic survival of these cells was determined after incubation with (111)In-NLS-trastuzumab, (111)In-trastuzumab, or trastuzumab (combined with or without MTX). Survival curves were analyzed according to the dose-response model, and the radiation-enhancement ratio was calculated from the survival curve parameters. RESULTS: The expression of HER2 was highest in SK-BR-3 cells (12.6 x 10(5) receptors/cell), compared with 231-H2N and TrR1 cells (6.1 x 10(5) and 5.1 x 10(5) receptors/cell, respectively), and lowest in MDA-MB-231 and TrR2 cells (0.4 x 10(5) and 0.6 x 10(5) receptors/cell, respectively). NLS peptides increased the nuclear uptake of (111)In-trastuzumab in MDA-MB-231, 231-H2N, TrR1, and TrR2 cells from 0.1%+/-0.01%, 2.5%+/-0.2%, 2.8%+/-0.7%, and 0.5%+/-0.1% to 0.5%+/-0.1%, 4.6%+/-0.1%, 5.2%+/-0.6%, and 1.5%+/-0.2%, respectively. The cytotoxicity of (111)In-NLS-trastuzumab on breast cancer cells was directly correlated with the HER2 expression densities of the cells. On a molar concentration basis, the effective concentration required to kill 50% of 231-H2N and TrR1 cells for (111)In-NLS-trastuzumab was 9- to 12-fold lower than for (111)In-trastuzumab and 16- to 77-fold lower than for trastuzumab. MDA-MB-231 and TrR2 cells were less sensitive to (111)In-NLS-trastuzumab or (111)In-trastuzumab, and both cell lines were completely insensitive to trastuzumab. The radiation-enhancement ratio induced by MTX for 231-H2N and TrR1 cells after exposure to (111)In-NLS-trastuzumab was 1.42 and 1.68, respectively. CONCLUSION: Targeted Auger electron radioimmunotherapy with (111)In-NLS-trastuzumab can overcome resistance to trastuzumab, and MTX can potently enhance the sensitivity of HER2-overexpressing breast cancer cells to the lethal Auger electrons emitted by this radiopharmaceutical.

Intracellular compartments, in particular the cytoplasm or nucleus, have generally been poorly accessible or inaccessible to radiolabeled biomolecules (e.g., monoclonal antibodies [mAbs], peptides, or oligonucleotides [ODNs]). However, recently cell-penetrating peptides (CPPs) and nuclear localizing peptide sequences (NLSs) have been shown to have the capability of inserting biomolecules into cells and transporting them to the cell nucleus. This discovery now presents intriguing new opportunities to design radiopharmaceuticals that could potentially probe, through imaging, the expression of key intracellular or intranuclear regulatory proteins that define the tumor phenotype, predict outcome, or act as sensitive reporters of response or resistance to treatment. CPPs could also more efficiently internalize radiolabeled antisense ODNs or peptide nucleic acids (PNAs) into tumor cells to enhance the sensitivity of imaging gene expression at the mRNA level. Perhaps one of the most exciting new developments to emerge is the use of NLS to route mAbs and peptides conjugated to nanometer-micrometer range Auger-electron-emitting radionuclides (e.g., (111)In) to the nucleus of cancer cells following their receptor-mediated internalization. In the nucleus, these electrons are highly potent in causing lethal DNA strand breaks. In some cases, NLSs are present naturally in peptide growth factors or their receptors, where they function to deliver internalized ligands to the nucleus, or alternatively, they can be introduced synthetically. This update reviews the properties of CPPs and NLS and focuses on their use for inserting radiolabeled biomolecules into cancer cells for imaging or targeted Auger electron radiotherapy of malignancies.

During the tenth season of excavations at Shahr-i Sokhta excavations continued in the Graveyard, and Craftsman's areas. This work revealed new graves, including one containing an artificial eye. Su...

We performed an observational pilot study of 18F-FLT PET/CT in pediatric lymphoma. Eight patients with equivocal 18F-FDG PET/CT underwent imaging with 18F-FLT PET/CT. No immediate adverse reactions to 18F-FLT were observed. Compared to 18F-FDG, 18F-FLT uptake was significantly higher in bone marrow and liver (18F-FLT SUV 8.6±0.6 and 5.0±0.3, versus 18F-FDG SUV 1.9±0.1 and 3.4±0.7, resp., p<0.05). In total, 15 lesions were evaluated with average 18F-FDG and 18F-FLT SUVs of 2.6±0.1 and 2.0±0.4, respectively. Nonspecific uptake in reactive lymph nodes and thymus was observed. Future studies to assess the clinical utility of 18F-FLT PET/CT in pediatric lymphoma are planned.

UNLABELLED: (111)In-DTPA-human epidermal growth factor ((111)In-DTPA-hEGF [DTPA is diethylenetriaminepentaacetic acid]) is an Auger electron-emitting radiopharmaceutical that targets EGF receptor (EGFR)-positive cancer. The purpose of this study was to determine the effect of EGFR inhibition by gefitinib on the internalization, nuclear translocation, and cytotoxicity of (111)In-DTPA-hEGF in EGFR-overexpressing MDA-MB-468 human breast cancer cells. METHODS: Western blot analysis was used to determine the optimum concentration of gefitinib to abolish EGFR activation. Internalization and nuclear translocation of fluorescein isothiocyanate-labeled hEGF were evaluated by confocal microscopy in MDA-MB-468 cells (1.3 x 10(6) EGFRs/cell) in the presence or absence of 1 microM gefitinib. The proportion of radioactivity partitioning into the cytoplasm and nucleus of MDA-MB-468 cells after incubation with (111)In-DTPA-hEGF for 24 h at 37 degrees C in the presence or absence of 1 microM gefitinib was measured by cell fractionation. DNA double-strand breaks caused by (111)In were quantified using the gamma-H2AX assay, and radiation-absorbed doses were estimated. Clonogenic survival assays were used to measure the cytotoxicity of (111)In-DTPA-hEGF alone or in combination with gefitinib. RESULTS: Gefitinib (1 microM) completely abolished EGFR phosphorylation in MDA-MB-468 cells. Internalization and nuclear translocation of fluorescein isothiocyanate-labeled EGF were not diminished in gefitinib-treated cells compared with controls. The proportion of internalized (111)In that localized in the nucleus was statistically significantly greater when (111)In-DTPA-hEGF was combined with gefitinib compared with (111)In-DTPA-hEGF alone (mean +/- SD: 26.0% +/- 5.5% vs. 14.6% +/- 4.0%, respectively; P < 0.05). Induction of gamma-H2AX foci was greater in MDA-MB-468 cells that were treated with (111)In-DTPA-hEGF (250 ng/mL, 1.5 MBq/mL) plus gefitinib (1 microM ) compared with those treated with (111)In-DTPA-hEGF alone (mean +/- SD: 35 +/- 4 vs. 24 +/- 5 foci per nucleus, respectively). In clonogenic assays, a significant reduction in the surviving fraction was observed when (111)In-DTPA-hEGF (5 ng/mL, 6 MBq/microg) was combined with gefitinib (1 microM ) compared with (111)In-DTPA-hEGF alone (42.9% +/- 5.7% vs. 22.9% +/- 3.6%, respectively; P < 0.01). CONCLUSION: The efficacy of (111)In-DTPA-hEGF depends on internalization and nuclear uptake of the radionuclide. Nuclear uptake, DNA damage, and cytotoxicity are enhanced when (111)In-DTPA-hEGF is combined with gefitinib. These results suggest a potential therapeutic role for peptide receptor radionuclide therapy in combination with tyrosine kinase inhibitors.

Highlight The integration of metabolic, molecular marker, and transcriptomic data from a segregating grapevine progeny provides novel insights into the genetic control of anthocyanin content and composition in ripe berries., In the last decade, great progress has been made in clarifying the main determinants of anthocyanin accumulation in grape berry skin. However, the molecular details of the fine variation among cultivars, which ultimately contributes to wine typicity, are still not completely understood. To shed light on this issue, the grapes of 170 F1 progeny from the cross ‘Syrah’×’Pinot Noir’ were characterized at the mature stage for the content of 15 anthocyanins during four growing seasons. This huge data set was used in combination with a dense genetic map to detect genomic regions controlling the anthocyanin pathway both at key enzymatic points and at particular branches. Genes putatively involved in fine tuning the global regulation of anthocyanin biosynthesis were identified by exploring the gene predictions in the QTL (quantitative trait locus) confidence intervals and their expression profile during berry development in offspring with contrasting anthocyanin accumulation. New information on some aspects which had scarcely been investigated so far, such as anthocyanin transport into the vacuole, or completely neglected, such as acylation, is provided. These genes represent a valuable resource in grapevine molecular-based breeding programmes to improve both fruit and wine quality and to tailor wine sensory properties according to consumer demand.