Extracapsular extension and tumor implants in lymph nodes with canine mammary carcinoma metastasis: Study of the impact on survival.

Regional lymph node status impacts survival in dogs with malignant mammary tumors. However, few studies have evaluated extracapsular extension and tumor implants in regional lymph node metastases in dogs with mammary carcinoma. Therefore, 84 cases of mammary carcinomas with metastases in inguinal and/or axillary lymph nodes from female dogs of different breeds and a total of 139 metastatic lymph nodes were evaluated by routine histological staining. Clinical and pathological characteristics of primary tumors as well as the presence of extracapsular extension and tumor implants in the lymph nodes were analyzed, in addition to survival. One to 5 lymph nodes were evaluated in each case. Extracapsular extension and tumor implants were present in 17.9% and 39.3% of cases, respectively. The simultaneous presence of extracapsular extension and tumor implants were associated with an increased risk of death (hazard ratio 10.46). In addition, "special type carcinomas", high histological grade (grade III), and presence of extracapsular extension associated with tumor implants were related to a worse prognosis and shorter survival times (p < 0.05). Based on these results, we highlight the importance of identifying extracapsular extension and tumor implants in dogs with metastatic mammary carcinomas, as they are associated with a higher risk of death and shorter survival.

Oregano Oil and Its Principal Component, Carvacrol, Inhibit HIV-1 Fusion into Target Cells.

Oregano essential oil has long been known for its health-promoting benefits. Here, we report its activity against viral replication. Oregano oil was found to specifically inhibit lentiviruses, such as human and simian immunodeficiency viruses (HIV and SIV), irrespective of virus tropism, but not hepatitis C virus, adenovirus 5 (ADV5), Zika virus, and influenza (H1N1) virus. Oregano oil's most abundant components, carvacrol and its isomer, thymol, were shown to block virus-target cell fusion while not perturbing other stages of the virus life cycle. We detected changes in virus particle density, suggesting that cholesterol depletion from the HIV-1 envelope membrane reduces virus entry. Furthermore, infection was rescued by adding exogenous cholesterol. The evolution of viral resistance to carvacrol supported this mechanism of action with the identification of mutations in the viral gp41 fusion protein that counteracted cholesterol depletion. In addition, resistance to carvacrol emerged later than typically observed for other clinically used drugs, strengthening its antiviral potential. Structure-activity relationship studies revealed key motifs of carvacrol and thymol required for HIV neutralization and identified previously unknown active analogs. Carvacrol was also shown to additively cooperate with antiretroviral therapy. In sum, oregano oil and improved carvacrol and thymol analogs could be considered to supplement current HIV therapeutics.IMPORTANCE Oregano essential oil has multiple benefits in traditional medicine, cosmetics, and food industries. Carvacrol and its analog, thymol, are well-described components of oregano oil. Here, we show that these compounds inhibit HIV-target cell fusion independently of viral tropism. Our results suggest that carvacrol and thymol alter the cholesterol content of the viral membrane, blocking HIV-1 entry into the target cell. Resistance to carvacrol has selected for viruses with mutations in the viral envelope glycoprotein, gp41. This protein is known for its interaction with cholesterol present in membrane lipid rafts. Together, these results demonstrate the potential of therapies targeting the viral envelope membrane, and oregano oil is a safe supplement to antiretrovirals, potentially delaying disease progression and resistance development.

The effect of omega-3 in temporomandibular joint synovial tissues of rats with induced arthritis: pilot study.

This study evaluated the effect of systemic administration of omega-3 on the expression of interleukins IL-1ß and IL-10 and tumour necrosis factor alpha (TNF-α) and on the thickness of cartilage in the temporomandibular joint (TMJ) inflammatory model induced by complete Freund's adjuvant (CFA). Thirty-two adult rats were divided equally into four groups: control, CFA (induced arthritis), and induced arthritis animals treated with dexamethasone or omega-3. The TMJs were then removed and assigned to histomorphometric analysis or immunoassay. The Kruskal-Wallis test with Dunn post hoc test was applied to the data; the significance level was set at 5%. IL-1ß levels (median; interquartile range) were higher (P<0.0001) in the CFA group (46.4 ng/ml; 39.4-53.3) than in the control group (1.81 ng/ml; 1.5-5.4), but there were no differences between the control, omega-3, and dexamethasone groups. TNF-α levels were also higher (P<0.0001) in the CFA group (122.7 ng/ml; 92.9-284.7) than in the control group (29.1 ng/ml; 23.7-31.3). IL-10 levels were lowest (P<0.0001) in the CFA group (73.5 ng/ml; 52.8-90.5), and no differences were found amongst the other groups. In conclusion, omega-3 successfully reduced the damage in the TMJ of induced arthritis rats. Further investigations are warranted to confirm whether the administration of omega-3 has a comparable effect to glucocorticoids in rheumatoid arthritis patients.

Data analysis software package for radionuclide standardization with a digital coincidence counting system.

The Nuclear Metrology Laboratory (LMN) - IPEN, São Paulo, Brazil - developed a Digital Coincidence System (DCS), based on the Coincidence Counting Methodology, in order to improve its capabilities in radionuclide primary-standardization. Digital process is implemented in two steps: data-acquisition (a set of measurements) and offline software data-analysis and calculation. The present work shows the basics of the data-acquisition unit (Software Coincidence System - SCS), describes the DCS' data-analysis process and the initial approaches chosen for the implementation of the software package (Coincidence Analyzing Task - CAT). 152Eu standardization, performed for DCS testing, software expansion and validation, is briefly discussed.

Publicaciones sobre hidroxicloroquina y remdesivir: actualización septiembre 2020 / Hydroxicloroquine and remdesivir publications: update September 2020

El debate científico sobre las terapéuticas eficaces y seguras para tratar la Covid-19 desde su comienzo en enero 2020 ha dado lugar a miles de publicaciones con sus implicaciones sociales y económicas respecto a fabricantes farmacéuticos, investigadores clínicos y publicaciones científicas. En este nuevo comentario se resumen las publicaciones de junio a septiembre de 2020 sobre los dos medicamentos más contrapuestos en muchos ámbitos clínicos: hidroxicloroquina y remdesivir

The scientific debate about the effective and safe therapies to treat the COVID-19 from its beginning in January 2020 has led to thousands of publications with their social and economic implications related to pharmaceutical manufacturers, clinical researchers and scientific publications. In this new comment, there are summarized the publications from June to September, 2020 about the two more opposing medications in many clinical settings: hydroxychloroquine and remdesivir

Estado de situación en Europa del Pharmaceutical Care / State of Pharmaceutical Care in Europe

En 2019 la revista Research in Social and Administrative Pharmacy, editada por ELSEVIER en California, con un factor de impacto de 2,873 para los últimos 5 años, ha publicado los resultados de un estudio sobre implementación de servicios de Atención Farmacéutica en las farmacias comunitarias de Europa en los años 2016-2017. Se realizan dos estudios independientes, ambos mediante envío de cuestionarios a profesionales seleccionados y relacionados con la farmacia comunitaria. El primero intenta conocer qué tipo de servicios cognitivos se ofrecen en cada país y su remuneración. Se publica en Febrero con resultados muy variados. El segundo, publicado en Noviembre, profundiza en la implementación de solo uno de aquéllos servicios: la Revisión de Medicación. Aprovecha la definición y clasificación que PCNE hace en 2016 de ese servicio y con ello consigue establecer un mapa de la implantación real en 34 países de Europa así como sus características profesionales y de remuneración. Parece de interés para España conocer estos datos ahora que algunas leyes autonómicas programan la implementación y coordinación de servicios de AF de acuerdo con las corporaciones farmacéuticas

The scientific magazine Research in Social and Administrative Pharmacy, edited by ELSEVIER in California, with an impact factor of 2,873 for the last 5 years published in 2019 the outcomes of a study about the implementation of Pharmaceutical Care Services in the community pharmacies of Europe over the years 2016 - 2017. Two independent studies were carried out, both through questionnaires sent to selected professionals linked to the community pharmacy. The first one tried to find out which kind of cognitive services are offered in each country and their remuneration. It was published in February with very varied outcomes. The second one, published in November, deepened in the implementation of only one of those services: the Medication Review. It took advantage of the definition and classification done by PCNE of that service and so it achieved to establish a map of the actual implementation in 34 European countries together with its professional characteristics and its remuneration. It seems interesting for Spain to know these data now that some autonomous laws plan the implementation and coordination of pharmaceutical care services in accordance with the pharmaceutical corporations

In vivo antimutagenic activity of the medicinal plants Pfaffia glomerata (Brazilian ginseng) and Ginkgo biloba.

Complementary and alternative therapies, including the use of medicinal plants, have become almost standard among the world's population. Pfaffia glomerata (PG), popularly known as Brazilian ginseng, is widely used as a restorer of vital functions, increasing mental balance, and is used for the treatment of diabetes and rheumatism. Ginkgo biloba (GB) is one of the oldest known gymnosperms, whose leaves are widely used for its potentiating action on the nervous system. The biological activities of these plants were determined on bone marrow cells of Wistar rats treated in vivo. For cytotoxic and mutagenic acute analysis, plant extracts were administered by gavage at concentrations of 0.15, 1.5, and 15 mg PG/mL water and 1, 2, and 3 mg GB/mL water. For antimutagenic analysis, plant extracts aqueous solution (PG, 1.5 mg/mL or GB, 2 mg/mL) were administered by gavage before (pretreatment), simultaneous to (simultaneous treatment), or after (post-treatment) the administration of cyclophosphamide (1.5 mg/mL, intraperitoneally). Both plant extracts have no cytotoxic or mutagenic potential, and they significantly reduce the percentage of chromosomal aberrations induced by the cyclophosphamide given simultaneously (PG, 87%; GB, 75%), pretreatment (PG, 98%, GB, 78%) and post-treatment (PG, 99%, GB, 75%). This beneficial antimutagenic property of the medicinal plants P. glomerata and G. biloba presented here, with no cytotoxic or mutagenic activity, can efficiently contribute to improvements in quality of life and recovery for people undergoing chemotherapeutic treatment, or those looking for health and preventive habits.

Synthetic Organotellurium Compounds Sensitize Drug-Resistant Candida albicans Clinical Isolates to Fluconazole.

Invasive Candida albicans infections are a serious health threat for immunocompromised individuals. Fluconazole is most commonly used to treat these infections, but resistance due to the overexpression of multidrug efflux pumps is of grave concern. This study evaluated the ability of five synthetic organotellurium compounds to reverse the fluconazole resistance of C. albicans clinical isolates. Compounds 1 to 4, at <10 µg/ml, ameliorated the fluconazole resistance of Saccharomyces cerevisiae strains overexpressing the major C. albicans multidrug efflux pumps Cdr1p and Mdr1p, whereas compound 5 only sensitized Mdr1p-overexpressing strains to fluconazole. Compounds 1 to 4 also inhibited efflux of the fluorescent substrate rhodamine 6G and the ATPase activity of Cdr1p, whereas all five of compounds 1 to 5 inhibited Nile red efflux by Mdr1p. Interestingly, all five compounds demonstrated synergy with fluconazole against efflux pump-overexpressing fluconazole-resistant C. albicans clinical isolates, isolate 95-142 overexpressing CDR1 and CDR2, isolate 96-25 overexpressing MDR1 and ERG11, and isolate 12-99 overexpressing CDR1, CDR2, MDR1, and ERG11 Overall, organotellurium compounds 1 and 2 were the most promising fluconazole chemosensitizers of fluconazole-resistant C. albicans isolates. Our data suggest that these novel organotellurium compounds inhibit pump efflux by two very important and distinct families of fungal multidrug efflux pumps: the ATP-binding cassette transporter Cdr1p and the major facilitator superfamily transporter Mdr1p.

First-in-human phase I study of copanlisib (BAY 80-6946), an intravenous pan-class I phosphatidylinositol 3-kinase inhibitor, in patients with advanced solid tumors and non-Hodgkin's lymphomas.

BACKGROUND: To evaluate the safety, tolerability, pharmacokinetics, and maximum tolerated dose (MTD) of copanlisib, a phosphatidylinositol 3-kinase inhibitor, in patients with advanced solid tumors or non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Phase I dose-escalation study including patients with advanced solid tumors or NHL, and a cohort of patients with type 2 diabetes mellitus. Patients received three weekly intravenous infusions of copanlisib per 28-day cycle over the dose range 0.1-1.2 mg/kg. Plasma copanlisib levels were analyzed for pharmacokinetics. Biomarker analysis included PIK3CA, KRAS, BRAF, and PTEN mutational status and PTEN immunohistochemistry. Whole-body [(18)F]-fluorodeoxyglucose positron emission tomography ((18)FDG-PET) was carried out at baseline and following the first dose to assess early pharmacodynamic effects. Plasma glucose and insulin levels were evaluated serially. RESULTS: Fifty-seven patients received treatment. The MTD was 0.8 mg/kg copanlisib. The most frequent treatment-related adverse events were nausea and transient hyperglycemia. Copanlisib exposure was dose-proportional with no accumulation; peak exposure positively correlated with transient hyperglycemia post-infusion. Sixteen of 20 patients treated at the MTD had reduced (18)FDG-PET uptake; 7 (33%) had a reduction >25%. One patient achieved a complete response (CR; endometrial carcinoma exhibiting both PIK3CA and PTEN mutations and complete PTEN loss) and two had a partial response (PR; both metastatic breast cancer). Among the nine NHL patients, all six with follicular lymphoma (FL) responded (one CR and five PRs) and one patient with diffuse large B-cell lymphoma had a PR by investigator assessment; two patients with FL who achieved CR (per post hoc independent radiologic review) were on treatment >3 years. CONCLUSION: Copanlisib, dosed intermittently on days 1, 8, and 15 of a 28-day cycle, was well tolerated and the MTD was determined to be 0.8 mg/kg. Copanlisib exhibited dose-proportional pharmacokinetics and promising anti-tumor activity, particularly in patients with NHL. CLINICALTRIALSGOV: NCT00962611; https://clinicaltrials.gov/ct2/show/NCT00962611.

Amphibian-killing chytrid in Brazil comprises both locally endemic and globally expanding populations.

Chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis (Bd), is the emerging infectious disease implicated in recent population declines and extinctions of amphibian species worldwide. Bd strains from regions of disease-associated amphibian decline to date have all belonged to a single, hypervirulent clonal genotype (Bd-GPL). However, earlier studies in the Atlantic Forest of southeastern Brazil detected a novel, putatively enzootic lineage (Bd-Brazil), and indicated hybridization between Bd-GPL and Bd-Brazil. Here, we characterize the spatial distribution and population history of these sympatric lineages in the Brazilian Atlantic Forest. To investigate the genetic structure of Bd in this region, we collected and genotyped Bd strains along a 2400-km transect of the Atlantic Forest. Bd-Brazil genotypes were restricted to a narrow geographic range in the southern Atlantic Forest, while Bd-GPL strains were widespread and largely geographically unstructured. Bd population genetics in this region support the hypothesis that the recently discovered Brazilian lineage is enzootic in the Atlantic Forest of Brazil and that Bd-GPL is a more recently expanded invasive. We collected additional hybrid isolates that demonstrate the recurrence of hybridization between panzootic and enzootic lineages, thereby confirming the existence of a hybrid zone in the Serra da Graciosa mountain range of Paraná State. Our field observations suggest that Bd-GPL may be more infective towards native Brazilian amphibians, and potentially more effective at dispersing across a fragmented landscape. We also provide further evidence of pathogen translocations mediated by the Brazilian ranaculture industry with implications for regulations and policies on global amphibian trade.

Insulin receptor isoforms: an integrated view focused on gestational diabetes mellitus.

The human insulin receptor (IR) exists in two isoforms that differ by the absence (IR-A) or the presence (IR-B) of a 12-amino acid segment encoded by exon 11. Both isoforms are functionally distinct regarding their binding affinities and intracellular signalling. However, the underlying mechanisms related to their cellular functions in several tissues are only partially understood. In this review, we summarize the current knowledge in this field regarding the alternative splicing of IR isoform, tissue-specific distribution and signalling both in physiology and disease, with an emphasis on the human placenta in gestational diabetes mellitus (GDM). Furthermore, we discuss the clinical relevance of IR isoforms highlighted by findings that show altered insulin signalling due to differential IR-A and IR-B expression in human placental endothelium in GDM pregnancies. Future research and clinical studies focused on the role of IR isoform signalling might provide novel therapeutic targets for treating GDM to improve the adverse maternal and neonatal outcomes.

Transcranial magnetic stimulation and aging: Effects on spatial learning and memory after sleep deprivation in Octodon degus.

The benefits of neuromodulatory procedures as a possible therapeutic application for cognitive rehabilitation have increased with the progress made in non-invasive modes of brain stimulation in aged-related disorders. Transcranial magnetic stimulation (TMS) is a non-invasive method used to examine multiple facets of the human brain and to ameliorate the impairment in cognition caused by Alzheimer's disease (AD). The present study was designed to evaluate how a chronic TMS treatment could improve learning and memory functions after sleep deprivation (SD) in old Octodon degus. SD was executed by gently handling to keep the animals awake throughout the night. Thirty young and twenty-four old O. degus females were divided in six groups (control, acute and chronic TMS treatment). Behavioral tests included; Radial Arm Maze (RAM), Barnes Maze (BM) and Novel Object Recognition (NOR). Although learning and memory functions improved in young animals with only one session of TMS treatment, a significant improvement in cognitive performance was seen in old animals after 4 and 7days of TMS, depending on the task that was performed. No side effects were observed following, which showed therapeutic potential for improving age-related cognitive performance.

Cognitive Impairment After Sleep Deprivation Rescued by Transcranial Magnetic Stimulation Application in Octodon degus.

Sleep is indispensable for maintaining regular daily life activities and is of fundamental physiological importance for cognitive performance. Sleep deprivation (SD) may affect learning capacity and the ability to form new memories, particularly with regard to hippocampus-dependent tasks. Transcranial magnetic stimulation (TMS) is a non-invasive procedure of electromagnetic induction that generates electric currents, activating nearby nerve cells in the stimulated cortical area. Several studies have looked into the potential therapeutic use of TMS. The present study was designed to evaluate how TMS could improve learning and memory functions following SD in Octodon degus. Thirty juvenile (18 months old) females were divided into three groups (control, acute, and chronic TMS treatment-with and without SD). TMS-treated groups were placed in plastic cylindrical cages designed to keep them immobile, while receiving head magnetic stimulation. SD was achieved by gently handling the animals to keep them awake during the night. Behavioral tests included radial arm maze (RAM), Barnes maze (BM), and novel object recognition. When TMS treatment was applied over several days, there was significant improvement of cognitive performance after SD, with no side effects. A single TMS session reduced the number of errors for the RAM test and improved latency and reduced errors for the BM test, which both evaluate spatial memory. Moreover, chronic TMS treatment brings about a significant improvement in both spatial and working memories.

Ribosome biogenesis dysfunction leads to p53-mediated apoptosis and goblet cell differentiation of mouse intestinal stem/progenitor cells.

Ribosome biogenesis is an essential cellular process. Its impairment is associated with developmental defects and increased risk of cancer. The in vivo cellular responses to defective ribosome biogenesis and the underlying molecular mechanisms are still incompletely understood. In particular, the consequences of impaired ribosome biogenesis within the intestinal epithelium in mammals have not been investigated so far. Here we adopted a genetic approach to investigate the role of Notchless (NLE), an essential actor of ribosome biogenesis, in the adult mouse intestinal lineage. Nle deficiency led to defects in the synthesis of large ribosomal subunit in crypts cells and resulted in the rapid elimination of intestinal stem cells and progenitors through distinct types of cellular responses, including apoptosis, cell cycle arrest and biased differentiation toward the goblet cell lineage. Similar observations were made using the rRNA transcription inhibitor CX-5461 on intestinal organoids culture. Importantly, we found that p53 activation was responsible for most of the cellular responses observed, including differentiation toward the goblet cell lineage. Moreover, we identify the goblet cell-specific marker Muc2 as a direct transcriptional target of p53. Nle-deficient ISCs and progenitors disappearance persisted in the absence of p53, underlying the existence of p53-independent cellular responses following defective ribosome biogenesis. Our data indicate that NLE is a crucial factor for intestinal homeostasis and provide new insights into how perturbations of ribosome biogenesis impact on cell fate decisions within the intestinal epithelium.

Mice engineered for an obligatory Mdm4 exon skipping express higher levels of the Mdm4-S isoform but exhibit increased p53 activity.

Mdm4, a protein related to the ubiquitin-ligase Mdm2, is an essential inhibitor of tumor suppressor protein p53. In both human and mouse cells, the Mdm4 gene encodes two major transcripts: one encodes the full-length oncoprotein (designated below as Mdm4-FL), whereas the other, resulting from a variant splicing that skips exon 6, encodes the shorter isoform Mdm4-S. Importantly, increased Mdm4-S mRNA levels were observed in several human cancers, and correlated with poor survival. However, the role of Mdm4-S in cancer progression remains controversial, because the Mdm4-S protein appeared to be a potent p53 inhibitor when overexpressed, but the splice variant also leads to a decrease in Mdm4-FL expression. To unambiguously determine the physiological impact of the Mdm4-S splice variant, we generated a mouse model with a targeted deletion of the Mdm4 exon 6, thereby creating an obligatory exon skipping. The mutant allele (Mdm4(ΔE6)) prevented the expression of Mdm4-FL, but also led to increased Mdm4-S mRNA levels. Mice homozygous for this allele died during embryonic development, but were rescued by a concomitant p53 deficiency. Furthermore in a hypomorphic p53(ΔP/ΔP) context, the Mdm4(ΔE6) allele led to p53 activation and delayed the growth of oncogene-induced tumors. We next determined the effect of Mdm4(+/ΔE6) heterozygosity in a hypermorphic p53(+/Δ31) genetic background, recently shown to be extremely sensitive to Mdm4 activity. Mdm4(+/ΔE6) p53(+/Δ31) pups were born, but suffered from aplastic anemia and died before weaning, again indicating an increased p53 activity. Our results demonstrate that the main effect of a skipping of Mdm4 exon 6 is not the synthesis of the Mdm4-S protein, but rather a decrease in Mdm4-FL expression. These and other data suggest that increased Mdm4-S mRNA levels might correlate with more aggressive cancers without encoding significant amounts of a potential oncoprotein. Hypotheses that may account for this apparent paradox are discussed.

Experimental strategies in performing value for cultivation and use experiments for the tobacco crop II: dimension of the experimental network.

In this study, we aimed to establish strategies for value for cultivation and use (VCU) experiments for the tobacco crop in the southern region of Brazil with respect to the number of environments used to assess tobacco lines. Trials of the Virginia (18 sites) and Burley (17 sites) varietal groups were conducted in the three states of the southern region of Brazil in the 2009-2010 crop season. The experiment was conducted in a completely randomized block design with four replications of 10 tobacco lines in the final stage of evaluation; the plots had 6 rows of 7 plants each, or 42 plants per plot. The cured leaf weight per hectare (kg/ha) was obtained. To evaluate stability, the ecovalence and additive main effects and multiplicative interaction models were adopted. In addition, joint analyses of variance were carried out considering different site numbers by simulating resampling. The site number ranged from 2 to 17 or 2 to 16, depending on the varietal group, and sites were selected at random without replacement. The process was repeated 2000 times for each number of sites. All analyses were performed using the R software. The results are very similar for both varietal groups. There is no advantage of using a large number of sites for VCU experiments in the southern region of Brazil because many sites contributed little to the interaction or did not discriminate the tobacco lines. Furthermore, the classification of the best lines is very similar to that obtained in the total number of evaluated sites.

Disintegration rate, gamma-ray emission probabilities and metastable half-life measurements of 67Ga.

The procedure for determining the (67)Ga disintegration rate by a primary method is described. The proposed triple 4πß-γ coincidence system consists of a thin window gas-flow 4π proportional counter (PC) coupled to a NaI(Tl) scintillator and a HPGe crystal. Independent pulse height and occurrence time information is provided for the three detector outputs by means of a Software Coincidence System. Separate spectrometry measurements with a n-type reversed electrode coaxial Ge detector (REGe) were performed for obtaining gamma-ray emission probabilities per decay. Accurate values of disintegration rate, gamma-ray emission probabilities and the metastable half-life were achieved.

The new facility for neutron tomography of IPEN-CNEN/SP and its potential to investigate hydrogenous substances.

A new facility for neutron tomography has been installed at the IEA-R1 nuclear research reactor of IPEN-CNEN/SP. A tomography can be obtained in 400 s and the spatial resolution in the image is 263 µm. The neutron dose per tomography, in the video camera used for image capture, is only 21 µSv, assures very few damages in its CCD sensor. Some selected objects were investigated and the obtained 3D images demonstrate the capability of the facility to investigate hydrogenous substances.

Mdm4 loss in mice expressing a p53 hypomorph alters tumor spectrum without improving survival.

The p53 pathway is inactivated in most human cancers, and its reactivation in tumors appears as a promising therapeutic strategy. Overexpression of Mdm4, a p53 negative regulator, occurs in a significant fraction of human cancers. Mouse models were used to evaluate the therapeutic potential of strategies against Mdm4, and encouraging results were obtained for tumor cells in which Mdm4 overexpression prevents wild-type p53 to exert its tumor suppressive functions. However, missense mutations in the p53 gene occur in about half of human cancers, and 15% of such mutations lead to the expression of a mutant protein that retains partial activity. In this report, we used mouse models to address the therapeutic potential of strategies against Mdm4 in tumors expressing an hypomorphic p53 mutant. We found that, in an Rb(+/-) background promoting pituitary and thyroid tumors, decreased Mdm4 levels improved the survival of mice expressing wild-type p53, but not that of mice expressing p53(ΔP), a p53 hypomorph lacking the proline-rich domain. Importantly, however, most Rb(+/-) p53(ΔP/ΔP) mice developped pituitary adenomas, but these tumors were rare in Rb(+/-) p53(ΔP/ΔP) Mdm4(-/-) animals, because Mdm4 loss led to increased p21 levels, a suppressor of pituitary tumor growth. On the contrary, Rb(+/-) p53(ΔP/ΔP) and Rb(+/-) p53(ΔP/ΔP) Mdm4(-/-) mice developped anaplastic thyroid carcinomas at equal frequencies. Importantly, wild-type p53 represses the Plk1 gene, which encodes a promising therapeutic target in anaplastic thyroid carcinomas, and this repression is improved when Mdm4 levels are decreased. On the opposite, p53(ΔP) is a mediocre transcriptional repressor that is not improved by Mdm4 loss. In sum, depending on the tumor type, strategies against Mdm4 that work in cells expressing wild-type p53 may not work in cells expressing an hypomorphic p53. Furthermore, p53-mediated transcriptional repression should be considered when evaluating strategies to reactivate p53 in tumors.

Experimental strategies in carrying out VCU for tobacco crop I: plot design and size.

We aimed to establish standards for tobacco Valor de Cultivo e Uso (VCU) in Brazil. We obtained information regarding the size and design of plots of two varietal groups of tobacco (Virginia and Burley). Ten inbred lines of each varietal group were evaluated in a randomized complete block design with four replications. The plot contained 42 plants with six rows of seven columns each. For each experiment plant, considering the position of the respective plant in the plot (row and column) as a reference, cured leaf weight (g/plant), total sugar content (%), and total alkaloid content (%) were determined. The maximum curvature of the variations in coefficients was estimated. Trials with the number of plants per plot ranging from 2 to 41 were simulated. The use of a border was not justified because the interactions between inbred lines x position in the plots were never significant, showing that the behavior of the inbred lines coincided with the different positions. The plant performance varied according to the column position in the plot. To lessen the effect of this factor, the use of plots with more than one row is recommended. Experimental precision, evaluated by the CV%, increased with an increase in plot size; nevertheless, the maximum curvature of the variation coefficient method showed no expressive increase in precision if the number of plants was greater than seven. The result in identification of the best inbred line, in terms of the size of each plot, coincided with the maximum curvature method.