Gene expression profile following an oral unsaturated fat load in abdominal obese subjects.

PURPOSE: Our aim was to evaluate the postprandial effect of an oral fat load test (OFLT) rich in unsaturated fatty acids on gene expression profile in peripheral blood mononuclear cells (PBMC) from subjects with abdominal obesity as an insulin resistance model and controls. METHODS: A total of 20 controls and 20 abdominal obese patients were studied. Metabolic parameters and oxidative stress markers were measured with standardized protocols. The whole gene expression at fasting state and after the OFLT (0, 4 and 8 h) was analysed using human HT-12-v4 expression beadchips, from Illumina. RESULTS: We found a significant decrease in plasma glucose, insulin and oxidative stress markers in abdominal obese patients and controls. We found beneficial metabolic postprandial gene expression in three genes: FKBP5, DDIT4 and DHRS9. Following an OFLT, the postprandial mRNA expression of FKBP5, and DDIT4 was downregulated while that of DHRS9 was overexpressed, both in nondiabetic normolipidemic subjects and in insulin-resistant subjects with abdominal obesity. CONCLUSIONS: Our results suggest that an OFLT rich in unsaturated fatty acids downregulates the expression of FKBP5, coding for the glucocorticoid receptor pathway, and that of DDIT4, involved in the oxidative stress response. These changes could favourably influence the insulin resistance and oxidative stress status in the postprandial state.

Immunohistochemical analysis of T-type calcium channels in acquired melanocytic naevi and melanoma.

BACKGROUND: Cutaneous malignant melanoma arises from transformed melanocytes de novo or from congenital or acquired melanocytic naevi. We have recently reported that T-type Ca2+ channels (TT-Cs) are upregulated in human melanoma and play an important role in cell proliferation. OBJECTIVES: To describe for the first time in formalin-fixed paraffin-embedded tissue the immunoexpression of TT-Cs in biopsies of normal skin, acquired melanocytic naevi and melanoma, in order to evaluate their role in melanomagenesis and/or tumour progression, their utility as prognostic markers and their possible use in targeted therapies. METHODS: Tissue samples from normal skin, melanocytic naevi and melanoma were subjected to immunohistochemistry for two TT-Cs (Cav3.1, Cav3.2); markers of proliferation (Ki67), the cell cycle (cyclin D1), hypoxia (Glut1), vascularization (CD31) and autophagy (LC3); BRAF V600E mutation (VE1) and phosphatase and tensin homologue (PTEN). Immunostaining was evaluated by histoscore. In silico analysis was used to assess the prognostic value of TT-C overexpression. RESULTS: TT-C immunoexpression increased gradually from normal skin to common naevi, dysplastic naevi and melanoma samples, but with differences in the distribution of both isoforms. Particularly, Cav3.2 expression was significantly higher in metastatic melanoma than in primary melanoma. Statistical correlation showed a linear interaction between PTEN loss/BRAF V600E/Cav3.1/LC3/ Ki67/cyclin D1/Cav3.2/Glut1. Disease-free survival (DFS) and overall survival correlated inversely with overexpression of Cav3.2. DFS also correlated inversely with overexpression of Cav3.1. CONCLUSIONS: TT-C immunoexpression on melanocytic neoplasms is consistent with our previous in vitro studies and appears to be related to tumour progression. TT-C upregulation can be considered as a prognostic marker using The Cancer Genome Atlas database. The high expression of Cav3.2 in metastatic melanoma encourages the investigation of the use of TT-C blockers in targeted therapies.

Morphological, cytogenetic, and molecular characterization of Arachis kuhlmannii Krapov. & W.C. Greg. (Leguminosae).

Arachis kuhlmannii occurs in Mato Grosso and Mato Grosso do Sul States, Brazil. Its area of occurrence partially overlaps with that of other species in the Arachis section. Because of their morphological similarities, these species are often mistaken one for another. This study aimed the correct classification of available accessions as Arachis kuhlmannii, or other species, and the characterization of similarities among accessions and Arachis hypogaea by morphological, cytogenetic, and molecular marker analyses. Thirty-eight accessions were used. Principal component analysis was used for morphological characterization, root tips for mitotic metaphase analysis, and RAPD markers for molecular characterization. Cluster analysis discriminated accessions with the A genome from the B genome. Cluster analysis based on molecular markers discriminated natural populations in a manner that correlated with geographical areas of the collection. Arachis cardenasii and A. hypogaea were isolated from other A-genome accessions. Cytogenetic analyses confirmed the existence of diagnostic characteristics that distinguish species with the A genome from those with the B genome. Results suggest the need for a taxonomic review of some species in the Arachis section, as we could not discriminate as distinct species all of the accessions identified as A. kuhlmannii, A. helodes, and A. simpsonii by using morphological, molecular, and cytogenetic markers.

Genetic diversity of the forage peanut in the Jequitinhonha, São Francisco, and Paranã River valleys of Brazil.

Arachis pintoi and A. repens are legumes with a high forage value that are used to feed ruminants in consortium systems. Not only do they increase the persistence and quality of pastures, they are also used for ornamental and green cover. The objective of this study was to analyze microsatellite markers in order to access the genetic diversity of 65 forage peanut germplasm accessions in the section Caulorrhizae of the genus Arachis in the Jequitinhonha, São Francisco and Paranã River valleys of Brazil. Fifty-seven accessions of A. pintoi and eight of A. repens were analyzed using 17 microsatellites, and the observed heterozygosity (HO), expected heterozygosity (HE), number of alleles per locus, discriminatory power, and polymorphism information content were all estimated. Ten loci (58.8%) were polymorphic, and 125 alleles were found in total. The HE ranged from 0.30 to 0.94, and HO values ranged from 0.03 to 0.88. By using Bayesian analysis, the accessions were genetically differentiated into three gene pools. Neither the unweighted pair group method with arithmetic mean nor a neighbor-joining analysis clustered samples into species, origin, or collection area. These results reveal a very weak genetic structure that does not form defined clusters, and that there is a high degree of similarity between the two species.

Role of primary care in the follow-up of patients with obstructive sleep apnoea undergoing CPAP treatment: a randomised controlled trial.

OBJECTIVE: To evaluate whether follow-up of patients with obstructive sleep apnoea (OSA) undergoing CPAP treatment could be performed in primary care (PC) settings. DESIGN: Non-inferiority, randomised, prospective controlled study. SETTINGS: Sleep unit (SU) at the University Hospital and in 8 PC units in Lleida, Spain. PARTICIPANTS: Patients with OSA were randomised to be followed up at the SU or PC units over a 6-month period. MAIN OUTCOMES MEASURED: The primary outcome was CPAP compliance at 6 months. The secondary outcomes were Epworth Sleep Scale (ESS) score, EuroQoL, patient satisfaction, body mass index (BMI), blood pressure and cost-effectiveness. RESULTS: We included 101 patients in PC ((mean±SD) apnoea-hypopnoea index (AHI) 50.8±22.9/h, age 56.2±11 years, 74% male) and 109 in the SU (AHI 51.4±24.4/h, age 55.8±11 years, 77% male)). The CPAP compliance was (mean (95% CI) 4.94 (4.47 to 5.5) vs 5.23 (4.79 to 5.66) h, p=0.18) in PC and SU groups, respectively. In the SU group, there were greater improvements in ESS scores (mean change 1.79, 95% CI +0.05 to +3.53, p=0.04) and patient satisfaction (-1.49, 95% CI -2.22 to -0.76); there was a significant mean difference in BMI between the groups (0.57, 95% CI +0.01 to +1.13, p=0.04). In the PC setting, there was a cost saving of 60%, with similar effectiveness, as well as a decrease in systolic blood pressure (-5.32; 95% CI -10.91 to +0.28, p=0.06). CONCLUSIONS: For patients with OSA, treatment provided in a PC setting did not result in worse CPAP compliance compared with a specialist model and was shown to be a cost-effective alternative. TRIAL REGISTRATION NUMBER: Clinical Trials NCT01918449.

Similar worldwide patterns in the sex pheromone signal and response in the oriental fruit moth, Grapholita molesta (Lepidoptera: Tortricidae).

The response of Grapholita molesta (Busck) males to three-component sex pheromone blends containing a 100% ratio of the major sex pheromone component, (Z)-8-dodecenyl acetate and a 10% ratio of (Z)-8-dodecenol, but with varying ratios of (E)-8-dodecenyl acetate (0.4, 5.4, 10.4, 30.4, and 100.1% E-blends) was tested with populations in eight stone and pome fruit orchards in Europe, Asia, and North and South America. Traps baited with the 5.4% E-blend caught significantly more males than traps with any other blend with all populations. Significantly more males were caught in traps baited with the 10.4% E-blend than in traps with the remaining blends, except with the 0.4% E-blend in Turkey. Significant differences in male moth catches occurred between the other blends with the 0.4>30.4% E-blend, and the 30.4>100.1% E-blend. Male moth catches with the 100.1% E-blend only differed from the hexane control in Chile. No apparent differences were noted to these blends in populations collected from pome or stone fruits. Flight tunnel assays to synthetic blends with a subset of populations were similar to the field results, but the breadth of the most attractive E-blends was wider. Flight tunnel assays also demonstrated a high level of male-female cross-attraction among field-collected populations. Female gland extracts from field-collected populations did not show any significant variation in their three-component blends. The only exceptions in these assays were that long-term laboratory populations were less responsive and attractive, and produced different blend ratios of the two minor components than recently collected field populations.

Crossability of Arachis valida and B genome Arachis species.

The peanut (Arachis hypogaea) is an important food crop in much of the tropical and semi-tropical parts of the world. The peanut is an allotetraploid with an AABB genome formula derived from diploids A. duranensis (A genome) and A. ipaënsis (B genome). The success of an introgression program that aims to improve cultivated varieties of the peanut depends on whether the chosen B genome species is homologous with the B genome of the peanut. While not directly involved in the origin of the peanut to the best of our knowledge, Arachis valida is a B genome species that could potentially be a bridge species or a source of new and different alleles, because of its resistance to diseases and pests. In this study, we investigated the crossability of A. valida with five other B genome species of section Arachis. Eight cross-combinations were made with A. valida and A. gregoryi, A. ipaënsis, A. magna, A. valida, and A. williamsii. Two hundred and forty pollinations were made yielding 61 fruit segments, 61 seeds, one abortion, and 24 hybrid plants. An analysis of the morphological characteristics and pollen viability confirmed that the plants were hybrids. Our results indicated that higher pollen viability of hybrid plants corresponded with higher affinity between parent plants used in crossings. This conclusion corroborates much of previous research carried out by many other authors in the past.

Complete asynapsis resulting in 2n pollen formation in Paspalum jesuiticum Parodi (Poaceae).

Cytological investigation revealed complete asynapsis during microsporogenesis in 2 wild accessions of Paspalum jesuiticum collected in distinct Brazilian regions. Both accessions were hexaploid (2n = 6x = 60) and 60 univalents could be counted at diakinesis. In this phase, the majority of meiocytes exhibited univalents with both chromatids. After alignment at the metaphase plate, the chromatids segregated to the poles. Only 1 meiotic division (equational) occurred, and after cytokinesis, a dyad with 2n microspores was formed. The genetic constitution of the 2n gametes was equivalent to that of first division restitution (FDR). Since recombination did not occur, 100% transmission of parental heterozygosity could be expected from the FDR 2n gametes. The meiotic behavior of both accessions suggested that they resulted from a recent natural hybridization event. The potential use of the 2n gametes in Paspalum breeding programs has been discussed.

Microsporogenesis in Paspalum conspersum Schrad. (Virgata group) with different ploidy levels.

Knowledge about the cytology and reproductive behavior of a species is indispensable for hybridization programs. This is especially true for species belonging to the genus Paspalum, among which apomixis and a wide range of ploidy levels are frequently found. Paspalum conspersum Schrad. is a robust and warm-season perennial bunchgrass native to South America. Previous studies have indicated that both tetraploid and hexaploid races exist in this species; however, only information related to tetraploids has been applied to another taxon. In this study, a cytological investigation in two Brazilian accessions collected in different regions revealed tetraploidy in the accession BRA-012823 (2n = 4x = 40), with chromosome pairing in bivalents and normal meiosis and tetrad formation, and pentaploidy (2n = 5x = 50) in the accession BRA-022748, which presented total asynapsis. In this latter accession, 50 univalents could be scored at diakinesis. After alignment at the metaphase plate, sister chromatids segregated to the poles. Only one meiotic division (equational) occurred, and after cytokinesis, 100% of the dyads that formed had 2n microspores. The meiotic behavior during microsporogenesis, which showed 10 delayed univalents to reach the metaphase plate, suggests that this accession is a recent natural hybrid constituted by a parental genome with 40 chromosomes and another with 10 chromosomes. The potential usage of these accessions in Paspalum breeding has been discussed.

Cuticular hydrocarbons discriminate cryptic Macrolophus species (Hemiptera: Miridae).

Macrolophus pygmaeus is commercially employed in the biological control of greenhouse and field vegetable pests. It is morphologically undistinguishable from the cryptic species M. melanotoma, and this interferes with the evaluation of the biological control activity of M. pygmaeus. We analysed the potential of cuticular hydrocarbon composition as a method to discriminate the two Macrolophus species. A third species, M. costalis, which is different from the other two species by having a dark spot at the tip of the scutellum, served as a control. Sex, diet and species, all had significant effects in the cuticular hydrocarbon profiles, but the variability associated to sex or diet was smaller than among species. Discriminant quadratic analysis of cuticular hydrocarbons confirmed the results of previous molecular genetic studies and showed, using cross-validation methods, that M. pygmaeus can be discriminated from M. costalis and M. melanotoma with prediction errors of 6.75% and 0%, respectively. Therefore, cuticular hydrocarbons can be used to separate M. pygmaeus from M. melanotoma reliably.

Post-pancreaticoduodenectomy hemorrhage. Incidence, diagnosis, and treatment.

BACKGROUND: Although mortality post-pancreaticoduodenectomy (PD) has decreased, morbidity rates continue to be high, ranging from 30% to 50%. Among complications, hemorrhage stands out; it is associated with high mortality and there is no standard management. The aim of the present study was to analyze the incidence, diagnosis, and treatment of hemorrhage post-cephalic PD at our center. METHODS: From January 2005 to December 2008, 107 PDs were performed. A retrospective review of characteristics of patients with postoperative hemorrhage was made from our prospective database. Demographic data, diagnosis, treatment (medical, laparotomy, interventional radiology), association with fistula (pancreatic or biliary), intra- or extraluminal hemorrhage, bleeding time (early or late), severity (moderate/severe), and mortality were analyzed. RESULTS: Eighteen patients (18/107; 16.82%) hemorrhaged after PD. Hemorrhage appeared early (< 24 h) in 4 of these 18 patients (22.2%), and it was severe in 13/18 (72%). Hemorrhage-related mortality was 11% (2/18) and hospital mortality was 22.2% (4/18). Arteriography was performed in 8/18 patients (44.4%) and was effective in 6/8 (75%); laparotomy was performed in 8/18 (44.4%). Re-bleeding occurred in 5 of these 18 patients after the first treatment (27.8%). An association between hemorrhage and fistula was observed. CONCLUSIONS: Hemorrhage after pancreatic resection must be considered a complication with relatively high mortality. Diagnosis should be established and treatment applied rapidly. Pancreatic and/or biliary fistulae were significantly associated with a higher risk of postoperative hemorrhage. Interventional radiology is a good therapeutic option.

Relationship between insulin resistance, inflammation and liver cell apoptosis in patients with severe obesity.

BACKGROUND: In obesity, insulin resistance appears frequently after activation of proinflammatory molecules. Caspase-generated cytokeratin-18 (CK-18) fragments are produced during the apoptosis of hepatic cells. The main objective in the present study is to investigate the relationship between insulin resistance and caspase-generated CK-18 fragments in patients with severe obesity. METHODS: Sixty-two patients selected for bariatric surgery were clinically studied (sex, age, weight, waist diameter, body mass index, arterial pressure and type 2 diabetes mellitus) and analytic parameters were measured in blood (glucose concentration, cholesterol, triglycerides, insulin, glycosylated hemoglobin, aspartate aminotransferase, alanine aminotransferase, high-sensitivity C-reactive protein, adiponectin, interleukin 6, interleukin 18 and CK-18 fragments). Patient group division was based on 70th percentile of insulin resistance as measured by homeostasis model assessment (HOMA) and also according to liver histology. RESULTS: Patients with greater insulin resistance (percentile > 70th) showed higher values of CK-18 fragments, interleukin 6 and transaminases. A positive correlation between the HOMA score, value of CK-18 fragments and triglyceride level was found. A correlation between CK-18 fragments with interleukin 6, triglycerides and transaminases was also observed. HOMA score and value of CK-18 fragments correlated with the degree of liver fibrosis. CONCLUSIONS: Greater degree of insulin resistance induces apoptosis of hepatic cells as measured by the serum levels of CK-18 fragments.

A probabilistic Poisson-based model to detect PRRSV recirculation using sow production records.

Porcine reproductive and respiratory syndrome (PRRS) is a viral disease associated with a decrease in the number of born alive piglets (NBA) and an increase in the number of lost piglets (NLP) per farrowing. Under practical conditions, it is critical to assess whether a farm is suffering PRRSV recirculation in the sow herd as soon as possible. The aim of this research work was to develop a new method to detect potential PRRSV recirculation in sow production farms. Sow reproductive performance records from one farm (farm T) were used to set up the method and records from ten additional farms (farms V1 to V10) were used for validation. A conditional Poisson model of NLP on NBA was proposed to fit the data. A three-step procedure was implemented to detect potential PRRSV recirculation: (i) computation of the maximum-likelihood estimates of the expected values of NBA and NLP in a PRRSV non-recirculating scenario; (ii) calculation, for each farrowing, of the p-value associated with the probability of jointly observing deviations towards decreased NBA and increased NLP. The detection of a potential PRRSV recirculation was based on (iii) the combined p-value resulting from weighing the p-values of the last N farrowings by the chi-square-inverse method. In order to gain specificity, a displacement on the expected non-recirculating NBA and NLP values was used for tuning purposes. With this approach, two PRRSV circulating periods were detected in farm T, which were confirmed with standard laboratorial diagnostic techniques. The method was subsequently validated in farms V1 to V10, where ten PRRSV-recirculating time episodes had been diagnosed. The method proposed here was able to detect the ten PRRSV recirculations using a relatively small set of contiguous farrowings, with only two mismatched weeks, one as a false negative, in farm V1, and one as a false positive, in farm V4. It is concluded that a conditional Poisson-based model of NLP on NBA can be a useful tool for routinely detecting PRRSV recirculation in sow herds.

Life expectancy and age-period-cohort effects: analysis and projections of mortality in Spain between 1977 and 2016.

OBJECTIVES: This study aimed to: (1) assess Spanish mortality trends between 1977 and 2001 and their impact on life expectancy; and (2) assess the differences in life expectancy between men and women for the period 2002-2016. STUDY DESIGN: Time trends study using age-period-cohort (APC) analysis. METHODS: A Bayesian APC model was fitted to describe Spanish mortality rates for the period 1977-2001 and to project Spanish mortality rates for 2002-2016. Life expectancy was predicted through Chiang's method using projected mortality rates. RESULTS: There was a significant cohort effect for Spanish mortality, showing a slight increase in mortality among men aged 20-39 years between 1986 and 1997 (birth cohorts 1940-1970). Life expectancy is expected to increase by approximately 0.5% in men and women between 1977 and 2016 (1 year per 5-year period). Life expectancy for males born between 2012 and 2016 will be 77.15 years, compared with 84.95 years for females born during the same period. CONCLUSIONS: The rising trend in mortality among the 1940-1970 cohorts may be due to the increased risk of avoidable causes of death related to acquired immunodeficiency syndrome, traffic accidents, and drug and alcohol abuse during the mid 1980s. The decline in mortality rates in recent years could lead to a mean increase in life expectancy of 1 year per 5-year period in both genders between 2002 and 2016. An increase in life expectancy for women and a levelling off for men is expected for age groups older than 79 years.

Unusual chromosome numbers in Paspalum L. (Poaceae: Paniceae) from Brazil.

Somatic chromosome numbers were determined for 20 new germplasm accessions of Paspalum, belonging to 17 species collected in Brazil. Chromosome number is reported for the first time for P. reduncum (2n = 18), P. cinerascens (2n = 20), P. cordatum (2n = 20), P. filgueirasii (2n = 24), P. ammodes (2n = 36), P. bicilium (2n = 40), P. heterotrichon (2n = 40), and P. burmanii (2n = 48). New cytotypes were confirmed for two germplasm accessions of P. carinatum (2n = 30) and P. trachycoleon (2n = 36), one of P. clavuliferum (2n = 40) and one of P. lanciflorum (2n = 40), indicating variability in these species. The remaining chromosome numbers reported here confirm previous counts. The unexpected chromosome numbers 2n = 18, 24, 36, and 48 in Paspalum species, which are usually shown to be multiples of 10, suggest that much more collection and cytogenetic characterization are necessary to assess the whole chromosomal and genomic multiplicity present in the genus, which seems to be much more diverse than currently thought to be.

[Trends in cancer incidence in Catalonia, 1985-2002]. / Evolución de la incidencia del cáncer en Cataluña, 1985-2002.

OBJECTIVE: To describe the secular trend of cancer incidence in Catalonia and to compare it with that observed in European regions. MATERIAL AND METHOD: Using data from the Tarragona and Gerona cancer registries, the number of cases, adjusted rates, and the annual percentages of change in cancer incidence in Catalonia for the period 1985-2002 were estimated. Cancer incidence trends in Catalonia, Spain, and four European regions were compared using variations in the adjusted rates between the periods 1993-1997 and 1998-2002. RESULTS: The number of invasive cancers increased from 15,773 in 1985 to 30,755 in 2002. The adjusted incidence rate showed an annual increase of 2.64% in males and of 1.81% in females. Almost all tumoral types showed an increasing trend. The most frequent malignant tumors in males were prostate, lung, colon and rectum, and urinary bladder tumors, showing increases of 8.74%, 1.67%, 3.47% and 4.32% respectively. The most frequent tumors in females were breast, colon and rectum and corpus uterine tumors, showing increases of 2.45%, 1.67% and 0.78%, respectively. In males, Catalonia showed lower annual incidence rates than the remaining European regions in 1985 and higher rates in 2002. In females, rates remained lower than in other European regions. CONCLUSIONS: In Catalonia, the number of incident cancers increased because of population growth and aging, greater exposure to risk factors and, for some cancer types, higher detection rates. Overall cancer incidence trends followed a similar pattern to those of southern Europe, with higher increases than in the remaining European regions, especially in males.

[Predictions of cancer incidence and mortality in Catalonia to 2015 by means of Bayesian models]. / Proyección de la incidencia y la mortalidad del cáncer en Cataluña hasta el año 2015 mediante un modelo bayesiano.

BACKGROUND AND OBJECTIVE: To perform cancer incidence and mortality projections in Catalonia for the period 2005-2019. To assess the projected increase in the incidence in 2015 compared with that in 2005, and to determine whether this increase is attributable to changes in risk or in demographics. POPULATION AND METHOD: Bayesian age-period-cohort models were fitted to age-specific rates for 1985-2004 to obtain the expected number of cases for the 5-year periods 2005-2009, 2010-2014 and 2015- 2019. Annual cases were estimated through a polynomial interpolation model. Incidence and mortality data were obtained from the Tarrragona and Gerona cancer registries, while population pyramids for the period 1985-2019 were obtained from the Catalan Institute of Statistics. RESULTS: In Catalonia, 27,438 cancer cases will be diagnosed among men and 19,986 among women in 2015, representing an increase in the number of cases diagnosed of 31% and 34%, respectively, when compared with those diagnosed in 2005 (20,999 and 14,141, respectively). In men, the increases attributable to risk, aging and demographic changes are 10%, 14% and 7%, respectively, whereas in women these changes are 6%, 13% and 15%. In the next decade, cancer mortality is expected to stabilize in men and to continue to decrease in women. Major increases in cancer incidence and mortality are expected among old age groups. CONCLUSIONS: The present study highlights the need to reorganize the resources and infrastructures required for cancer control and care, taking into account the predicted burden of oncology patients.

Circulating tumor cells (CTCs) and cytokeratin 19 (CK19) mRNA as prognostic factors in heavily pretreated patients with metastatic breast cancer.

INTRODUCTION: Circulating tumor cell (CTC) count and cytokeratin 19 (CK19) mRNA expression have a prognostic value for patients with metastatic breast cancer (MBC), but their clinical utility remains controversial. We studied CTC count and CK19 mRNA expression in the peripheral blood samples from heavily pretreated patients with MBC and their correlations with prognosis and response to the subsequent line of therapy. METHODS: This prospective observational study included 67 consecutive patients with MBC who were on progression to systemic therapy, and criteria for a new line of systemic treatment were proposed outside a clinical trial. CTC counts and CK19 mRNA expression were measured by the CellSearch® and RT-PCR methods, respectively, before and after the first cycle of treatment. Progression-free survival (PFS) was defined as the time elapsed between the initiation of the treatment and either the date of clinical or radiological tumor progression or death from any cause or the last follow-up. Cox proportional hazards regression model was used to assess the univariate prognostic value of CTC and CK19 mRNA expression on PFS and Kaplan-Meier estimates. A multivariate Cox model was also used to additionally account for phenotype and visceral disease. RESULTS: The mean age was 60 (range 35-86) years, and the average number of previous treatments was 3 (range 1-10); 42 patients (62.6%) were ER+ and 38 patients (56.7%) had visceral disease. The median PFS rate was 8 months (95% CI: 3.7-8.2). Univariate analyses showed a significant effect of the initial value of CK19 mRNA expression (HR = 2.00; 95% CI: 1.05-3.8; p = 0.03) and for the second value of CTC (HR = 2.18; 95% CI: 1.22-3.9; p = 0.009) but did not reach statistical significance for the initial value of CTC and the second value of CK19 mRNA expression. The estimated PFS rates at 6 and 12 months were 75% and 31% for patients with a low initial value of CK19 mRNA expression and 36% and 10% for those with a high initial value of CK19 mRNA expression, respectively (p: 0.022). Further, the estimated PFS rates at 6 and 12 months were 86% and 65% for patients with a low second value of CTC and 76% and 47% for those with a high second value of CTC, respectively (p: 0.004). In the multivariate analysis adjusted for phenotype, visceral disease, and the last treatment performed, only the effect of the second value of CTC remained significant (HR = 2.7, p = 0.004). CONCLUSIONS: CK19 mRNA expression and CTC count appeared clinically meaningful in pretreated patients with MBC, even when adjusted for phenotype and visceral disease involvement. These results support the use of CK19 and CTC as relevant biomarkers for predicting clinical response in MBC.

Topical Analgesia with Lidocaine Plus Diclofenac Decreases Pain in Benign Anorectal Surgery: Randomized, Double-blind, and Controlled Clinical Trial.

OBJECTIVE: The aim of this study is to evaluate the efficacy and safety of a topical formulation containing lidocaine plus diclofenac (CLIFE1) compared to lidocaine (CLIFE2), to decrease pain in benign anorectal surgery (BARS) to date not evaluated. More than 50% of patients undergoing BARS, especially hemorrhoidectomy, suffer from moderate and severe postoperative pain. This remains an unresolved problem that could be addressed with the new CLIFE1 topical treatment. METHODS: A multicenter, randomized double-blind, active-controlled parallel-group superiority trial, was conducted in two Spanish hospitals. Patients undergoing BARS (hemorrhoids, anal fistula and anal fissure) were randomized at the end of surgery at a 1:1 ratio to receive first dose either CLIFE1 (n = 60) or CLIFE2 (n = 60) anorectal topical treatment, and after every 12 h for the first three postoperative days and once a day from the fourth to sixth. The primary outcome was average of pain decrease after topical treatment, measured with visual analogue scale (VAS) by the patients themselves, the evening in the surgery day and four times daily for the first three postoperative days. RESULTS: The results of 120 patients included out of 150 selected undergoing BARS show a decrease in pain after CLIFE1 topical treatment (7.47 ± 13.09) greater than with CLIFE2 (4.38 ± 6.75), difference -3.21 95% CI (-5.75; -0.68), p = 0.008, decreasing significantly postoperative pain ( ≥ 9 mm, VAS) in 35% of patients undergoing benign anorectal surgery, compared to 18.33 % treated with lidocaine. CONCLUSIONS: The CLIFE1 topical treatment shows better analgesic efficacy than CLIFE2 in BARS.

Genetic variability of Brazilian Indian landraces of Arachis hypogaea L.

The Kayabi Indians who inhabit the Xingu Indigenous Park, located in West Central Brazil, have grown and managed peanuts for a long time. A great number of landraces are being maintained by these tribes and some of this germplasm has morphological traits that exceed the variation described in the taxonomic literature. Here, we analyzed the genetic variability of these landraces using a set of microsatellite markers. The analysis showed that, in general, the indigenous samples grouped according to the villages where they were collected. The microsatellite markers used in the present study detected high levels of genetic variation. Similarity groups, genetically distant from each other, were formed, allowing a more efficient use of the existing genetic variability. The present study also showed that these materials can extend the genetic variability available for peanut-breeding programs. Additionally, the microsatellite markers revealed a large dissimilarity among germplasm accessions representing Arachis hypogaea varieties so far included in the same subspecies fastigiata (aequatoriana + peruviana vs fastigiata + vulgaris), a subject that deserves further investigation. Finally, the Xingu Indigenous Park proved to be an important center of diversity for peanut.