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The knowledge about the biological events that regulate lipid metabolism in oocytes and embryos in buffalo is scarce. Lipogenesis, lipolysis, transport and oxidation of fatty acids (FAs) occur in gametes and embryonic cells of all mammalian species, as an intrinsic component of energy metabolism. In oocytes and cumulus cells, degradation of lipids is responsible for the production of ATP that is essential for the metabolic processes that lead to oocyte maturation in in vivo and in vitro culture conditions. Similarly, throughout embryo development, blastomeres have the capacity to use exogenous and/or endogenous lipid reserves to serve as an energy source necessary for early embryonic development. In addition, supplementation of culture media with L-carnitine to promote lipid metabolism during in vitro oocyte maturation and early embryonic development leads to an improved embryo quality. The limited scientific evidence available in buffalo indicates there is relatively greater oocyte lipid content as compared with many other species that undergoes a dynamic distribution during folliculogenesis and follicle maturation and that has a positive effect on oocyte maturation and embryo development when there is L-carnitine supplementation of the media. Advances in the understanding of the biological peculiarities of lipid metabolism, and the consequences of its alteration on the quality of buffalo gametes and embryos, therefore, are necessary to design specific culture media and laboratory procedures as a strategy to increase in vitro-derived embryo production rates., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Transvaginal follicular aspiration technique together with in vitro embryo production are the biotechnological alternatives currently available to support genetic improvement breeding programs in buffalo species. However, aspects related to animal management, lack of knowledge of the metabolic needs and biochemical peculiarities of gametes and embryos, as well as the reproductive physiology characteristics have hampered progress in the results. Despite the low availability of good quality oocytes collected after OPU in donors as a physiological characteristic of buffalo species, high rates of oocyte maturation, modest embryo cleavage, blastocyst production and pregnancy rates after transvaginal embryo transfer in recipients could be obtained in buffalo in vitro embryo production programs. The results of implementing an in vitro embryo production program in buffaloes in the northern region of Pará state, Brazil, and results published by other groups demonstrate the feasibility of implementing this biotechnology in the routine of breeding programs. Nevertheless, in order to achieve better and consistent results, it is necessary to deepen the knowledge on the peculiarities of reproductive biology in this specie. Selection of donor animals based on ovarian size and ovarian follicular reserve and on the rate of blastocyst production is presented as an effective alternative to increase the efficiency of the in vitro embryo production technique applied to the buffalo species., (Copyright © The Author(s). Published by CBRA.)

This study aimed to determinate the chemical composition and evaluate the antimicrobial and antioxidant activity of the essential oil obtained from leaves of V. gardneriana. The Vitex gardneriana leaves's were hydrodistilled to obtain the essential oil and the chemical composition determined by GC/MS analysis. The antimicrobial activities were determined by microdilution method. The activity of essential oil on biofilm was evaluated by quantification of total biomass and enumeration of biofilm-entrapped viable cells. The antioxidant activity was assessed by DPPH free radical assay, ferrous ion chelating assay, ferric-reducing antioxidant power and β-carotene bleaching assay. Furthermore, the essential oil was tested on viability of health human, animal cells and the microcrustacean Artemia sp. The essential oil showed high content of sesquiterpenes and very low content of monoterpenes. Regarding activity on planktonic cells, the essential oil reduced the growth of the all species tested but showed MIC values only to S. aureus (0.31%). In general, the essential oil reduced significantly the biofilm biomass and the number of viable cells of bacteria and yeasts, mainly on biofilm formation. The essential oil showed a potential antioxidant activity, mainly on β-carotene oxidation. Moreover, the essential oil reduced the cell viability of murine fibroblasts but not show viability reduction of human keratinocytes. Furthermore, the oil not show toxicity against the microcrustacean. Thus, the essential oil from V. gardneriana leaves may be considered as an important alternative against biofilms formed by bacteria and yeasts related to infections, as well as a natural antioxidant and non-toxic substance on human cells., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The dynamics of dengue transmission are multifactorial and involve socioeconomic, ecological, and environmental aspects, the latter being closely related to local climatic conditions that affect the vector's reproductive cycle. Climate depends in turn on tropical oceanic mechanisms such as phases of El Niño/La Niña over the Pacific. The study contributes to this discussion and reports on the correlations between the Multivariate ENSO Index (MEI) in the Pacific and the number of reported dengue cases in seven state capitals in the Brazilian Amazon from 2001 to 2012. The study also analyzes the seasonality pattern (quarterly mean values) in dengue cases throughout the region. Evidence that El Niño/La Niña causes a decrease versus increase in the local rainfall pattern is consistent with the lower versus higher number of reported dengue cases in most of the state capitals in the Amazon, a result proven by the statistically significant negative correlations seen in Manaus (Amazonas), São Luís (Maranhão), Belém (Pará) and Palmas (Tocantins). The 12-years means (2001-2012) revealed the presence of pronounced seasonality in dengue incidence in the majority of the state capitals, with sharp peaks from January to March [Rio Branco (Acre), Manaus, Belém and Palmas] and from April to June (São Luís), corresponding to 50-70% of the annual total. State capitals farther north [Boa Vista (Roraima) and Macapá (Amapá)] showed dengue reporting in all quarters of the year, with no pronounced seasonality.

Purpose: To assess functional outcomes in patients undergoing total knee arthroplasty (TKA) without previous corrective osteotomy for treatment of knee osteoarthritis associated with extra-articular deformity., Methods: From January to December 2016, patients with knee osteoarthritis with extra-articular deformities who presented for preoperative assessment before TKA were evaluated prospectively. Physical and radiological characteristics were documented pre- and postoperatively., Results: TKA was performed in 33 knees; 25 were considered for analysis. The mean age was 65.2 years (range, 48-79 years). Sixteen deformities were secondary to fractures and nine to failed osteotomies. The mean Knee Society Score (KSS) improved from 27.1 pre-operatively to 68.7 post-operatively (p = 0.000). Pre-operative mechanical axis ranged from 32° varus (negative) to 26° valgus. After correction, 20 knees were within 3° (varus or valgus) of mechanical alignment., Conclusion: In patients with extra-articular deformities, TKA with asymmetric intra-articular resection and ligament balancing can relieve pain and realign the mechanical axis of the lower limb.

We present new constraints on the dark matter-induced annual modulation signal using 1.7 years of COSINE-100 data with a total exposure of 97.7 kg yr. The COSINE-100 experiment, consisting of 106 kg of NaI(Tl) target material, is designed to carry out a model-independent test of DAMA/LIBRA's claim of WIMP discovery by searching for the same annual modulation signal using the same NaI(Tl) target. The crystal data show a 2.7  cpd/kg/keV background rate on average in the 2-6 keV energy region of interest. Using a χ-squared minimization method we observe best fit values for modulation amplitude and phase of 0.0092±0.0067  cpd/kg/keV and 127.2±45.9  d, respectively.

Wound healing can be delayed following colonization and infection with the common bacterium Pseudomonas aeruginosa . While multiple therapies are used for their treatment, these are ineffective, expensive, and labour-intensive. Thus, there is an enormous unmet need for the treatment of infected wounds. Cinnamaldehyde, the major component of cinnamon oil, is well known for its antimicrobial properties. Herein, we investigated the effects of sub-inhibitory concentrations of cinnamaldehyde in the virulence of P. aeruginosa . We also assessed its healing potential in P. aeruginosa -infected mouse skin wounds and the mechanisms involved in this response. Sub-inhibitory concentrations of cinnamaldehyde reduced P. aeruginosa metabolic rate and its ability to form biofilm and to cause haemolysis. Daily topical application of cinnamaldehyde on P. aeruginosa -infected skin wounds reduced tissue bacterial load and promoted faster healing. Lower interleukin-17 (IL-17), vascular endothelial growth factor (VEGF) and nitric oxide levels were detected in cinnamaldehyde-treated wound samples. Blockage of transient receptor potential ankyrin 1, the pharmacological target of cinnamaldehyde, abrogated its healing activity and partially reversed the inhibitory actions of this compound on VEGF and IL-17 generation. We suggest that topical application of sub-inhibitory concentrations of cinnamaldehyde may represent an interesting approach to improve the healing of P. aeruginosa -infected skin wounds.

The pathovar viticola of Xanthomonas citri causes bacterial canker of grapevine. This disease was first recorded in India in 1972, and later in Brazil in 1998, where its distribution is currently restricted to the northeastern region. A multilocus sequence analysis (MLSA) based on seven housekeeping genes and a multilocus variable number of tandem repeat analysis (MLVA) with eight loci were performed in order to assess the genetic relatedness among strains from India and Brazil. Strains isolated in India from three related pathovars affecting Vitaceae species and pathogenic strains isolated from Amaranthus sp. found in bacterial canker-infected vineyards in Brazil were also included. MLSA revealed lack of diversity in all seven genes and grouped grapevine and Amaranthus strains in a monophyletic group in X. citri . The VNTR (variable number of tandem repeat) typing scheme conducted on 107 strains detected 101 haplotypes. The total number of alleles per locus ranged from 5 to 12. A minimum spanning tree (MST) showed that Brazilian strains were clearly separated from Indian strains, which showed unique alleles at three loci. The two strains isolated from symptomatic Amaranthus sp. presented unique alleles at two loci. STRUCTURE analyses revealed three groups congruent with MST and a fourth group with strains from India and Brazil. Admixture among populations were observed in all groups. MST, STRUCTURE and e-BURST analyses showed that the strains collected in 1998 belong to two distinct groups, with predicted founder genotypes from two different vineyards in the same region. This suggest that one introduction of grape planting materials contaminated with genetically distinct strains took place, which was followed by pathogen adaptation. Genome sequencing of one Brazilian strain confirmed typical attributes of pathogenic xanthomonads and allowed the design of a complementary VNTR typing scheme dedicated to X. citri pv. viticola that will allow further epidemiological survey of this genetically monomorphic pathovar.

Introduction: Staphylococcus aureus represents the most common etiologic agent of purulent infections, affecting humans and animals. Escherichia coli is one of the principal causes of infectious diseases, mainly diarrheal diseases due to enterotoxin action. There are many reports indicating that these bacteria are multidrug-resistant (MDR) pathogens., Objective: In this study, we investigated the antimicrobial and modulatory activities of 5-hydroxy-3,7,4'-trimethoxyflavone (VG.EF.CLII) against E. coli and S. aureus strains., Methods: 5-Hydroxy-3,7,4'-trimethoxyflavone was isolated from Vitex gardneriana Schauer leaves and structurally characterized using nuclear magnetic resonance. The antibacterial effect of VG.EF.CLII and modulation of antibiotic activity, both determined by minimum inhibitory concentration, were assessed using microtiter plates., Results: VG.EF.CLII showed bacterial growth inhibition at concentrations ≤512 μg/mL, and synergistic effects were observed for the modulation of two distinct antibiotic classes (the fluoroquinolone norfloxacin and the aminoglycoside gentamicin)., Conclusion: 5-Hydroxy-3,7,4'-trimethoxyflavone isolated from V. gardneriana showed promising antimicrobial activity against MDR bacterial strains S. aureus 358 and E. coli 27 when associated with the antibiotics norfloxacin and gentamicin. Therefore, this natural product can contribute to the control of bacterial resistance.

Grapevine bacterial canker, which is caused by Xanthomonas campestris pv. viticola, is one of the most important grapevine diseases in the northeastern region of Brazil. This disease causes severe damage and represents a high potential risk to the development of Brazilian viticulture. In turn, pigmented isolates pathogenic to cashew plant, making cashew fruit unfit for sale, also have been detected in Northeastern Brazil. Given that the taxonomic position of these bacteria is unclear, the multilocus sequence analysis (MLSA) technique, average nucleotide identity (ANI) values and tetranucleotide frequency correlation coefficients (TETRA) were used to analyze their phylogenetic relationship in relation to other Xanthomonas species. X. campestris pv. viticola was closely related to X. citri pv. mangiferaeindicae (repetitive-polymerase chain reaction [rep-PCR], MLSA, and ANI) and X. citri subsp. citri (MLSA and ANI). Pigmented isolates pathogenic to cashew plant were closely related to X. citri pv. anacardii (rep-PCR, MLSA, ANI, and TETRA). The results obtained in this study support the emendation of the description of X. citri pv. anacardii to include pigmented isolates of Xanthomonas pathogenic to cashew plant. In addition, the reclassification of X. campestris pv. viticola as X. citri pv. viticola comb. nov. is suggested.

Despite the novel diagnostic methods and therapies implemented in oncology, the number of patients that succumb by the cancer remains high globally. Currently studies point out that 20-25% of all human malignancies are related to micro-organism infections. Among these cancer-related pathogens, the human papillomavirus (HPV) has a prominent position, since the virus is responsible for about 30% of all infectious agent-related cancers. Thus, an amount of cancers could be avoided by means prophylactic and/or therapeutic measures. However, these measures required a holistic comprehension about HPV-related cancer biology. Based on this, this review aims to summarize the last evidences of HPV on cancer biology (from initiation to metastasis), focus on molecular and biochemical deregulations associated with viral infection, and discuss the viral etiology in different malignancies., (Copyright © 2018. Published by Elsevier Masson SAS.)

This study describes the first antibiofilm and antibacterial screening for plants from Caatinga against Ralstonia solanacearum, a causal agent of bacterial wilt that presents serious difficulties in control. There were prepared 22 aqueous extracts of plants collected in the Vale do Catimbau-PE, Brazil. The potential antibacterial activity was evaluated by absorbance in OD 600 and the antibiofilm activity through the crystal violet method, both of them performed in microplate against isolates of R. solanacearum biofilm formers. The results of the screening showed that Jacaranda rugosa presented antimicrobial activity higher than 90%, while Harpochilus neesianus and Myroxylon peruiferum presented antibiofilm activity higher than 50% for all tested isolates. However, Croton heliotropiifolius showed both the activities, being thus very promising for application in the control of this phytopathogen. The search for viable alternatives to the development of new bioactive compounds safe for the environment, humans, and animals from an adverse and scarce environment such as the Caatinga and encouraged us to find plants that produce effective metabolites against phytopathogenic microorganisms. This in vitro screening is important to guide the development of new products in addition to guide research studies of bioactive compounds.

Brazilian Northeast is the world's largest producer of Agave sisalana Perrine for the supply of the sisal fiber. About 95% of plant biomass, which comprise the mucilage and sisal juice, is considered a waste residual is discarded in the soil. However, the sisal juice is rich in steroidal saponins, which exhibits different pharmacological properties. Despite this, natural products are not necessarily safe. Based on this, this study analyzed the antioxidant, cytotoxic and mutagenic potential of three extracts derived from acid hydrolysis (AHAS), dried precipitate (DPAS) and hexanic of A. sisalana (HAS). These analyses were performed by in vitro and in vivo methods, using Vero cells, human lymphocytes and mice. Results showed that AHAS 50 and 100 can be considered a useful antineoplastic candidate due to their antioxidant and cytotoxic activity, with no genotoxic/clastogenic potential in Vero cells and mice. Although the comet assay in human lymphocytes has showed that the AHAS 25, AHAS 50 and AHAS 100 can lead to DNA breaks, these extracts did not promote DNA damages in mice bone marrow. Considering the different mutagenic responses obtained with the different methods employed, this study suggest that the metabolizing pathways can produce by-products harmful to health. For this reason, it is mandatory to analyze the mutagenic potential by both in vitro and in vivo techniques, using cells derived from different species and origins., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

The bacterium Xanthomonas citri pv. anacardii is the agent of angular leaf spot of the cashew tree ( Anacardium occidentale L.). The complete genome sequencing of the strain IBSBF2579 was done on an Illumina HiSeq 2500 platform. The de novo assembly of the X. citri pv. anacardii strain IBSBF2579 genome yielded 133 contigs, with a size of 5,329,247 bp and a G+C content of 64.03%. The prediction was performed by GeneMarkS and the automatic annotation by Rapid Annotations using Subsystems Technology (RAST), with 4,406 identified genes., (Copyright © 2018 Silva Junior et al.)

The COSINE-100 dark matter search experiment is an array of NaI(Tl) crystal detectors located in the Yangyang Underground Laboratory (Y2L). To understand measured backgrounds in the NaI(Tl) crystals we have performed Monte Carlo simulations using the Geant4 toolkit and developed background models for each crystal that consider contributions from both internal and external sources, including cosmogenic nuclides. The background models are based on comparisons of measurement data with Monte Carlo simulations that are guided by a campaign of material assays and are used to evaluate backgrounds and identify their sources. The average background level for the six crystals (70 kg total mass) that are studied is 3.5 counts/day/keV/kg in the (2-6) keV energy interval. The dominant contributors in this energy region are found to be 210 Pb and 3 H.

Here, we report the complete 5.3-Mb genome sequence of Xanthomonas campestris pv. viticola (CCRMXCV 80), which causes grapevine ( Vitis vinifera L.) bacterial canker. Genome data will improve our understanding of the strain's comparative genomics and epidemiology, and help to further define plant protection and quarantine procedures., (Copyright © 2017 Lima et al.)

We report here the complete genome sequences of two Ralstonia pseudosolanacearum strains, isolated from the warm northeast region of Brazil. They display divergent (compatible versus incompatible) interactions with the resistant tomato line Hawaii 7996. Polymorphisms were detected in a subset of effector genes that might be associated with these contrasting phenotypes., (Copyright © 2017 Albuquerque et al.)

This study estimated the reference evapotranspiration rate (ETo) for the Itacaiúnas River Watershed (IRW), Eastern Amazonia, and measured the accuracy of eight empirical equations: Penman-Monteith (PM), Priestley-Taylor (PT), Hargreaves and Samani (HS), Camargo (CAM), Thornthwaite (TH), Hamon (HM), Kharrufa (KF) and Turc (TC) using monthly data from 1980 to 2013. In addition, it verifies the regional applicability to the IRW using a for the Marabá-PA station. The methods TC and PM (FAO56) presented the best results, which demonstrate that radiation and higher temperatures are the dominant drivers in the Evapotranspiration process, while relative humidity and wind speed have a much smaller impact. The temporal and spatial variability of ETo for IRW show has strong seasonality, increasing during the dry season and decreasing during the rainy season. The statistical analyses at 1% level of significance, indicates that there is no correlation of the residuals between the dry and rainy seasons, and test of the physical parameters such as mean temperature, solar radiation and relative air humidity explains the variations of ETo.

Bacterial resistance to the available marketed drugs has prompted the search of novel therapies; especially in regards of anti-virulence strategies that aim to make bacteria less pathogenic and/or decrease their probability to become resistant to therapy. Cinnamaldehyde is widely known for its antibacterial properties through mechanisms that include the interaction of this compound with bacterial cell walls. However, only a handful of studies have addressed its effects on bacterial virulence, especially when tested at sub-inhibitory concentrations. Herein, we show for the first time that cinnamaldehyde is bactericidal against Staphylococcus aureus and Enterococcus faecalis multidrug resistant strains and does not promote bacterial tolerance. Cinnamaldehyde actions were stronger on S. aureus as it was able to inhibit its hemolytic activity on human erythrocytes and reduce its adherence to latex. Furthermore, cinnamaldehyde enhanced the serum-dependent lysis of S. aureus . In vivo testing of cinnamaldehyde in Galleria mellonella larvae infected with S. aureus , showed this compound improves larvae survival whilst diminishing bacterial load in their hemolymph. We suggest that cinnamaldehyde may represent an alternative therapy to control S. aureus -induced bacterial infections as it presents the ability to reduce bacterial virulence/survival without promoting an adaptive phenotype.

This study evaluated the effects of a polymeric biosurfactant produced by Trichosporon montevideense CLOA72 in the adhesion of Candida albicans and Candida krusei cells to human buccal epithelial cells and its interference in biofilm formation by these strains. The biofilm inhibition by biosurfactant (25 mg/mL) in C. krusei and C. albicans in polystyrene was reduced up to 79.5 and 85 %, respectively. In addition, the zeta potential and hydrodynamic diameter of the yeasts altered as a function of the biosurfactant concentration added to the cell suspension. The changes in the cell surface characteristics and the interface modification can contribute to the inhibition of the initial adherence of yeasts cells to the surface. In addition, the analyses of the biofilm matrix and planktonic cell surfaces demonstrated differences in carbohydrate and protein concentrations for the two studied strains, which may contribute to the modulation of cell adhesion or consolidation of biofilms, especially in C. krusei. This study suggests a possible application of the of CLOA72 biosurfactant in inhibiting the adhesion and formation of biofilms on biological surfaces by yeasts of the Candida genus.

Long-term human-induced impacts have significantly changed the Amazonian landscape. The most dramatic land cover and land use (LCLU) changes began in the early 1970s with the establishment of the Trans-Amazon Highway and large government projects associated with the expansion of agricultural settlement and cattle ranching, which cleared significant tropical forest cover in the areas of new and accelerated human development. Taking the changes in the LCLU over the past four decades as a basis, this study aims to determine the consequences of land cover (forest and savanna) and land use (pasturelands, mining and urban) changes on the hydroclimatology of the Itacaiúnas River watershed area of the located in the southeastern Amazon region. We analyzed a multi-decadal Landsat dataset from 1973, 1984, 1994, 2004 and 2013 and a 40-yr time series of water discharge from the Itacaiúnas River, as well as air temperature and relative humidity data over this drainage area for the same period. We employed standard Landsat image processing techniques in conjunction with a geographic object-based image analysis and multi-resolution classification approach. With the goal of detecting possible long-term trends, non-parametric Mann-Kendall test was applied, based on a Sen slope estimator on a 40-yr annual PREC, TMED and RH time series, considering the spatial average of the entire watershed. In the 1970s, the region was entirely covered by forest (99%) and savanna (∼0.3%). Four decades later, only ∼48% of the tropical forest remains, while pasturelands occupy approximately 50% of the watershed area. Moreover, in protected areas, nearly 97% of the tropical forest remains conserved, while the forest cover of non-protected areas is quite fragmented and, consequently, unevenly distributed, covering an area of only 30%. Based on observational data analysis, there is evidence that the conversion of forest cover to extensive and homogeneous pasturelands was accompanied by systematic modifications to the hydroclimatology cycle of the Itacaiúnas watershed, thus highlighting drier environmental conditions due to a rise in the region's air temperature, a decrease in the relative humidity, and an increase in river discharge., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Objective: to identify the reasons and motivations why family members search treatment for the drug addicted., Method: descriptive qualitative research, developed in 2012 and 2013, in a Drug Addicts Rehabilitation Unit of Parana State, Brazil. A total of 19 semi-structured interviews were conducted with the drug addicts' family members in treatment. The results were analyzed based on the Transtheoretical Model of Behavior Change and organized in thematic categories according with qualitative data analysis., Results: the search for treatment for drug addicts occurred: in the pre-contemplation stage influenced by external factors; in the contemplation stage both for ambivalence and behavioral changes needs; in the action stage by awareness of drug addiction and also professional help needs; and in the maintenance stage because of the non-conservation of behavioral changes., Conclusion: an evaluation of motivational stages in the beginning of treatment is required for expansion of success possibilities in the rehabilitation process.

Physical, chemical and biological agents can act in the DNA, resulting in mutation involved in cancer. Thus, genotoxic tests are required by regulatory agencies in order to evaluate potential risk of cancer. Among these tests, the comet assay (CA) and micronucleus assay (MNA) are the most commonly used. However, there are different protocols and recommendations already published. This is the first review, after the inclusion of CA in S2R1 guidance and OECD 489, which summarizes the main technical recommendations of both CA and MNA., (Copyright © 2015. Published by Elsevier Masson SAS.)

Background: Finding the best extraction method of proteins from lysed cells is the key step for detection and identification in all proteomics applications. These are important to complement the knowledge about the mechanisms of interaction between plants and phytopathogens causing major economic losses. To develop an optimized extraction protocol, strains of Acidovorax citrulli, Pectobacterium carotovorum subsp. carotovorum and Ralstonia solanacearum were used as representative cells in the study of phytopathogenic bacteria. This study aims to compare four different protein extraction methods, including: Trizol, Phenol, Centrifugation and Lysis in order to determine which are more suitable for proteomic studies using as parameters the quantity and quality of extracted proteins observed in two-dimensional gels., Results: The bacteria studied showed different results among the tested methods. The Lysis method was more efficient for P. carotovorum subsp. carotovorum and R. solanacearum phytobacteria, as well as simple and fast, while for A. citrulli, the Centrifugation method was the best. This evaluation is based on results obtained in polyacrylamide gels that presented a greater abundance of spots and clearer and more consistent strips as detected by two-dimensional gels., Conclusions: These results attest to the adequacy of these proteins extraction methods for proteomic studies.

Empirical orthogonal functions (EOF) and composites analysis were employed on pentad data in order to investigate the tropical atmospheric-ocean patterns over the Atlantic Ocean and the spatial-temporal characteristics of the rainfall in eastern Amazon during the peak of the rainy season (February to April). The EOF results evidenced that the Intertropical Convergence Zone (ITCZ) is the main rainfall-producing system in eastern Amazon during the rainy season. Conditions associated with the southward SST gradient in the intertropical Atlantic formed the dynamic patterns that favored the position of the ITCZ to south of the equator, thus explaining the predominance of positive precipitation anomalies in eastern Amazon, especially in the state of Maranhão and northeastern Pará during the February and April months.

Bovine papillomavirus (BPV) is considered a useful model to study HPV oncogenic process. BPV interacts with the host chromatin, resulting in DNA damage, which is attributed to E5, E6, and E7 viral oncoproteins activity. However, the oncogenic mechanisms of BPV E6 oncoprotein per se remain unknown. This study aimed to evaluate the mutagenic potential of Bos taurus papillomavirus type 1 (BPV-1) E6 recombinant oncoprotein by the cytokinesis-block micronucleus assay (CBMNA) and comet assay (CA). Peripheral blood samples of five calves were collected. Samples were subjected to molecular diagnosis, which did not reveal presence of BPV sequences. Samples were treated with 1 μg/mL of BPV-1 E6 oncoprotein and 50 μg/mL of cyclophosphamide (positive control). Negative controls were not submitted to any treatment. The samples were submitted to the CBMNA and CA. The results showed that BPV E6 oncoprotein induces clastogenesis per se, which is indicative of genomic instability. These results allowed better understanding the mechanism of cancer promotion associated with the BPV E6 oncoprotein and revealed that this oncoprotein can induce carcinogenesis per se. E6 recombinant oncoprotein has been suggested as a possible vaccine candidate. Results pointed out that BPV E6 recombinant oncoprotein modifications are required to use it as vaccine.

The epidemic situation of Moko disease-causing strains in Latin America and Brazil is unclear. Thirty-seven Ralstonia solanacearum strains from Brazil that cause the Moko disease on banana and heliconia plants were sampled and phylogenetically typed using the endoglucanase (egl) and DNA repair (mutS) genes according to the phylotype and sequevar classification. All of the strains belonged to phylotype II and a portion of the strains was typed as the Moko disease-related sequevars IIA-6 and IIA-24. Nevertheless, two unsuspected sequevars also harbored the Moko disease-causing strains IIA-41 and IIB-25, and a new sequevar was described and named IIA-53. All of the strains were pathogenic to banana and some of the strains of sequevars IIA-6, IIA-24, and IIA-41 were also pathogenic to tomato. The Moko disease-causing strains from sequevar IIB-25 were pathogenic to potato but not to tomato. These results highlight the high diversity of strains of Moko in Brazil, reinforce the efficiency of the egl gene to reveal relationships among these strains, and contribute to a better understanding of the diversity of paraphyletic Moko disease-causing strains of the R. solanacearum species complex, where the following seven distinct genetic clusters have been described: IIA-6, IIA-24, IIA-41, IIA-53, IIB-3, IIB-4, and IIB-25.

Obesity is one of the most important nutritional disorders, and can be currently considered as an epidemic. Although there are few weight reduction drugs available on the market, some new drug candidates have been proposed, including Cordia ecalyculata, a Brazilian plant with anorectic properties, and Spirulina maxima, a cyanobacterium with antioxidant and anti-genotoxic activity. In this study, we evaluated the mutagenic potential of C. ecalyculata at doses of 150, 300, and 500 mg/kg alone and in association with S. maxima at doses of 75, 150, and 250 mg/kg, respectively, through an in vivo micronucleus test, using mice of both sexes, and an in vitro micronucleus test and comet assay, using human peripheral blood. For all tests, cyclophosphamide was used as a positive control. The results showed that treatment of 300 mg/kg C. ecalyculata and the combination treatment of 500 mg/kg C. ecalyculata with 250 mg/kg S. maxima resulted in anorectic effects. The mutagenic tests did not reveal any clastogenic or genotoxic activity for any treatment, indicating that these candidates could be marketed as weight-reduction drugs. Moreover, the drugs contain chemo-preventive substances that can protect against tumorigenesis, which has been associated with obesity.

In this study we evaluate the efficacy of five vaccine formulations containing different combinations of proteins (FimH; leukotoxin, LKT; and pyolysin, PLO) and/or inactivated whole cells (Escherichia coli, Fusobacterium necrophorum, and Trueperella pyogenes) in preventing postpartum uterine diseases. Inactivated whole cells were produced using two genetically distinct strains of each bacterial species (E. coli, F. necrophorum, and T. pyogenes). FimH and PLO subunits were produced using recombinant protein expression, and LKT was recovered from culturing a wild F. necrophorum strain. Three subcutaneous vaccines were formulated: Vaccine 1 was composed of inactivated bacterial whole cells and proteins; Vaccine 2 was composed of proteins only; and Vaccine 3 was composed of inactivated bacterial whole cells only. Two intravaginal vaccines were formulated: Vaccine 4 was composed of inactivated bacterial whole cells and proteins; and Vaccine 5 was composed of PLO and LKT. To evaluate vaccine efficacy, a randomized clinical trial was conducted at a commercial dairy farm; 371 spring heifers were allocated randomly into one of six different treatments groups: control, Vaccine 1, Vaccine 2, Vaccine 3, Vaccine 4 and Vaccine 5. Late pregnant heifers assigned to one of the vaccine groups were each vaccinated twice: at 230 and 260 days of pregnancy. When vaccines were evaluated grouped as subcutaneous and intravaginal, the subcutaneous ones were found to significantly reduce the incidence of puerperal metritis. Additionally, subcutaneous vaccination significantly reduced rectal temperature at 6±1 days in milk. Reproduction was improved for cows that received subcutaneous vaccines. In general, vaccination induced a significant increase in serum IgG titers against all antigens, with subcutaneous vaccination again being more effective. In conclusion, subcutaneous vaccination with inactivated bacterial components and/or protein subunits of E. coli, F. necrophorum and T. pyogenes can prevent puerperal metritis during the first lactation of dairy cows, leading to improved reproduction.

Objective: Kidney dysfunction is a major complication in the postoperative cardiac surgery setting. Operative risk factors for its development are cardiopulmonary bypass, anemia, antifibrinolytic drugs and blood transfusion. The objective of this study was to identify the risk factors for developing kidney dysfunction in patients undergoing cardiac surgery., Methods: Ninety-seven patients were studied and 84 were analyzed. The sample was stratified into two groups. A serum creatinine higher than 30% compared to the preoperative period was considered for the kidney dysfunction group (n=9; 10.71%). There also was a control group when the increase in serum creatinine remained lower than 30% (n=75; 89.28%)., Results: It was observed that intraoperative transfusion of fresh frozen plasma in the control group was 2.05 ± 0.78 units and 3.80 ± 2.16 units in the kidney dysfunction group with P= 0.032., Conclusion: It was possible to associate that fresh frozen plasma transfusion is a risk factor for postoperative kidney dysfunction after cardiovascular surgery.

The export of cashew (Anacardium occidentale) nuts generates millions of dollars for the Brazilian economy annually. However, production may be limited by the occurrence of diseases that affect cashew trees, such as Xanthomonas spot and angular leaf spot, which are caused by pigmented strains of Xanthomonas and Xanthomonas citri pv. anacardii, respectively. Thirty-one pigmented strains of Xanthomonas were characterized for phenotypic, pathogenic, and molecular attributes. These strains were similar to X. citri pv. anacardii in phenotypical characteristics, sensitivity to antibiotics and copper compounds used in agriculture, epidemiology, and repetitive sequence-based polymerase chain reaction (rep-PCR) profiles. When inoculated into Brazilian pepper, cashew, mango, and hog plum seedlings, the pigmented strains of Xanthomonas and X. citri pv. anacardii produced similar symptoms. However, the pigmented strains of Xanthomonas were more aggressive toward cashew plants than toward the other hosts tested, which confirms their specificity. We conclude that pigmented strains of Xanthomonas are very aggressive on cashew trees and should not be considered casual pathogens of these hosts. Moreover, based on our results from rep-PCR and IS1595-PCR amplification, we suggest that these strains constitute a variant of X. citri pv. anacardii.

Immunosuppression has been reported to occur during active visceral leishmaniasis and some factors such as the cytokine profile may be involved in this process. In the mouse model of cutaneous leishmaniasis using Leishmania (Leishmania) major, the Th1 response is related to protection while the Th2 response is related to disease progression. However, in hamsters, which are considered to be an excellent model for the study of visceral leishmaniasis, this dichotomy is not observed. Using outbred 45- to 60-day-old (140 to 150 g) male hamsters infected intraperitoneally with 2 x 10(7) L. (L.) chagasi amastigotes, we evaluated the immune response of spleen cells and the production of cytokines. We used 3 to 7 hamsters per group evaluated. We detected a preserved response to concanavalin A measured by index of proliferation during all periods of infection studied, while a proliferative response to Leishmania antigen was detected only at 48 and 72 h post-infection. Messenger RNA from cytokines type 1 (IL-2, TNF-α, IFN-γ) and type 2 (IL-4, IL-10 and TGF-β) detected by reverse transcriptase polymerase chain reaction and produced by spleen cells showed no qualitative difference between control non-infected hamsters and infected hamsters during any period of infection evaluated. Cytokines were measured by the DNA band intensity on agarose gel using the Image Lab 1D L340 software with no differences observed. In conclusion, the present results showed an antigen-dependent immunosuppression in hamsters with active visceral leishmaniasis that was not related to the cytokine profile.

Wild felids and canids are usually the main predators in the food chains where they dwell and are almost invisible to behavior and ecology researchers. Due to their grooming behavior, they tend to swallow shed hair, which shows up in the feces. DNA found in hair shafts can be used in molecular studies that can unravel, for instance, genetic variability, reproductive mode and family structure, and in some species, it is even possible to estimate migration and dispersion rates in given populations. First, however, DNA must be extracted from hair. We extracted successfully and dependably hair shaft DNA from eight wild Brazilian felids, ocelot, margay, oncilla, Geoffroy's cat, pampas cat, jaguarundi, puma, and jaguar, as well as the domestic cat and from three wild Brazilian canids, maned wolf, crab-eating fox, and hoary fox, as well as the domestic dog. Hair samples came mostly from feces collected at the São Paulo Zoo and were also gathered from non-sedated pet or from recently dead wild animals and were also collected from museum specimens. Fractions of hair samples were stained before DNA extraction, while most samples were not. Our extraction protocol is based on a feather DNA extraction technique, based in the phenol:chloroform:isoamyl alcohol general method, with proteinase K as digestive enzyme.

Two unusual monoterpene indole alkaloids, stachyoside (1) and nor-methyl-23-oxo-correantoside (2), have been isolated from the aerial parts of Psychotria stachyoides. The structural elucidation of both compounds was performed by the aid of HRESIMS, FT-IR, and 1D- and 2D-NMR techniques including COSY, HSQC, HMBC, and NOESY., (2010 John Wiley & Sons, Ltd.)

The effects of five extraction solvents and their mixtures on the yield of metabolites in crude and fractionated extracts of Annona muricata L. leaves were investigated by direct comparison. Extraction media were prepared using simplex centroid mixtures of ethanol, ethyl acetate, dichloromethane, acetone, and chloroform. The effects of the mobile phase solvent strength and the analysis wavelength on the chromatographic separation were also investigated. Solvent mixtures rather than pure solvents were found to be the most efficient extractors for the different fractions. The results indicated that the mobile phase composed of methanol/acetonitrile/water (26:27:47 v/v/v) was most suitable for the basic fraction analysis at 254 nm, whereas the mobile phase composed of methanol/acetonitrile/water (35:35:30 v/v/v) was the most adequate for the organic fraction analysis at 254 nm. The results indicated that the chromatographic profiles and number of peaks were affected by the mobile phase strength and analysis wavelength.

PDI, a redox chaperone, is involved in host cell uptake of bacteria/viruses, phagosome formation, and vascular NADPH oxidase regulation. PDI involvement in phagocyte infection by parasites has been poorly explored. Here, we investigated the role of PDI in in vitro infection of J774 macrophages by amastigote and promastigote forms of the protozoan Leishmania chagasi and assessed whether PDI associates with the macrophage NADPH oxidase complex. Promastigote but not amastigote phagocytosis was inhibited significantly by macrophage incubation with thiol/PDI inhibitors DTNB, bacitracin, phenylarsine oxide, and neutralizing PDI antibody in a parasite redox-dependent way. Binding assays indicate that PDI preferentially mediates parasite internalization. Bref-A, an ER-Golgi-disrupting agent, prevented PDI concentration in an enriched macrophage membrane fraction and promoted a significant decrease in infection. Promastigote phagocytosis was increased further by macrophage overexpression of wild-type PDI and decreased upon transfection with an antisense PDI plasmid or PDI siRNA. At later stages of infection, PDI physically interacted with L. chagasi, as revealed by immunoprecipitation data. Promastigote uptake was inhibited consistently by macrophage preincubation with catalase. Additionally, loss- or gain-of-function experiments indicated that PMA-driven NADPH oxidase activation correlated directly with PDI expression levels. Close association between PDI and the p22phox NADPH oxidase subunit was shown by confocal colocalization and coimmunoprecipitation. These results provide evidence that PDI not only associates with phagocyte NADPH oxidase but also that PDI is crucial for efficient macrophage infection by L. chagasi.

Small molecule drugs are relatively effective in working on 'drugable' targets such as GPCRs, ion channels, kinases, proteases, etc but ineffective at blocking protein-protein interactions that represent an emerging class of 'nondrugable' central nervous system (CNS) targets. This article provides an overview of novel therapeutic modalities such as biologics (in particular antibodies) and emerging oligonucleotide therapeutics such as antisense, small-interfering RNA, and aptamers. Their key properties, overall strengths and limitations, and their utility as tools for target validation are presented. In addition, issues with regard to CNS targets as it relates to the blood-brain barrier penetration are discussed. Finally, examples of their application as therapeutics for the treatment of pain and some neurological disorders such as Alzheimer's disease, multiple sclerosis, Huntington's disease, and Parkinson's disease are provided.

Congenital or developmental deformities of the chest rarely cause severe functional problems in the manner of larger anomalies of the limbs. Poland syndrome is known as a variety of associated anomalies centered on the trunk and upper limbs. The authors report the case of a 28-year-old female with absence of the pectoralis major muscle and depression of the left anterior thoracic wall (sunken chest). She presented with hypoplasia of the papillary-areolar complex; her upper left limb was normal. Seven years after undergoing placement of a left-side breast prosthesis, she requested replacement of the implant because she was dissatisfied with its shape. Chest radiographs showed a chest deformity and deviation of the mediastinum to the right. After surgically removing the prosthesis, an area of costal reabsorption on the left side was seen at its site, with exposure of the parietal pericardium. The prosthesis was replaced with another of greater volume and the patient was referred to the thoracic surgery service. No cases of regional bone resorption secondary to the presence of a breast prosthesis are reported in the literature. However, it is known that compression results in bone absorption and subsequent bone remodeling. The authors therefore believe that greater attention to this possibility is needed in cases of breast prosthesis replacement among patients with Poland syndrome, by means of specific imaging studies such as computed tomography.

Objective: The objective of this study was to evaluate the disinfection degree of dentine caused by the use of diode laser after biomechanical procedures., Study Design: Thirty teeth were sectioned and roots were autoclaved and incubated for 4 weeks with a suspension of Enterococcus faecalis. The specimens were randomly divided into 3 groups (n = 10): G1, instrumented with rotary files, irrigated with 0.5% sodium hypochlorite and 17% EDTA-T, and then irradiated by 830-nm diode laser at 3 W; G2, the same procedures as G1 but without laser irradiation; and G3, irrigation with saline solution (control). Dentin samples of each third were collected with carbide burs and aliquots were sowed to count viable cells., Results: The disinfection degree achieved was 100% in G1 and 98.39% in G2, when compared to the control group (G3)., Conclusion: Diode laser irradiation provided increased disinfection of the deep radicular dentin in the parameters and samples tested.

A systematic literature review was conducted to investigate the effects of physical activity during pregnancy on selected maternal-child health outcomes. The search included articles published from 1980 to 2005 in the MEDLINE and LILACS databases using key words such as physical activity, physical exercise, pregnancy, and gestation. The methodological quality of 37 selected articles was evaluated. It appears to be a consensus that some light-to-moderate physical activity is not a risk factor and may even be considered a protective factor for some outcomes. However, some studies found an association between specific activities (e.g., climbing stairs or standing for long periods) and inadequate birth weight, prematurity, and miscarriage. Few studies found an association between physical activity and maternal weight gain, mode of delivery, or fetal development. Further research is needed to fill these gaps and provide guidelines on the intensity, duration, and frequency of physical activity during pregnancy.

The microsatellite loci FCA045, FCA077, FCA008, and FCA096 are highly variable molecular markers which were used to determine the genetic diversity in 148 captive Leopardus sp. The PCR-amplified products of microsatellite loci were characterized in ABI Prism 310 Genetic Analyzer. Allele numbers, heterozygosity, polymorphism information content, exclusive allele number, and shared alleles were calculated. Sixty-five alleles were found and their sizes ranged from 116 to 216 bp in four microsatellite loci. The heterozygosity ranged from 0.36 to 0.81 in Leopardus pardalis, 0.57 to 0.67 in L. tigrinus and 0.80 to 0.92 in L. wiedii. The polymorphism information content was from 0.80 to 0.88 in L. pardalis, 0.76 to 0.88 in L. tigrinus and 0.77 to 0.90 in L. wiedii. The margay (L. wiedii) showed the highest index of polymorphism among the three species in this study. These results imply that microsatellite DNA markers can help in the study of the genetic diversity of Leopardus specimens.

Apical sealing is essential for the success of paraendodontic surgery, so any procedure that may favor an adequate sealing of the apical remainder should be performed. The purpose of this study was to evaluate the influence of diode laser irradiation on the apical sealing of root-end cavities with MTA retrofillings. Root canals in twenty extracted human teeth were shaped with K-files and filled with gutta-percha. The apexes were cut off and root-end preparations were performed. The roots were divided randomly in 2 groups. Group 1 (ten specimens) was retrofilled with MTA. Group 2 was irradiated with diode laser, with 1 W for 20 seconds, on the apical surface and root end cavity before retrofilling with MTA. The specimens had their external surfaces impermeabilized with cyanoacrylate, except for the apical surface, and were then immersed in 1% rhodamine B dye for 72 h and placed in plaster stone. After that, the specimens were submitted to longitudinal abrasion until half of the root remained. The linear dye leakage was observed in these mid-roots between the root canal wall and retrofilling. The linear dye leakage was measured with Image Lab software, and the results were statistically analyzed with Student's t test. There were no statistically significant differences between the two groups (p > 0.05). The diode laser irradiation did not improve the apical sealing of MTA retrofillings under the conditions of this in vitro study.

The volatile composition of the essential oils from leaves and roots of Eupatorium betonicaeforme (D.C.) Baker was analyzed by GC-MS. A total of 12 compounds were identified. beta-Caryophyllene (12.4-41.7%), alpha-humulene (11.7-14.6%), gamma-muurolene (10.4-19.0%), bicyclogermacrene (15.0-17.5%), 2,2-dimethyl-6-vinylchroman-4-one (10.3-25.5%), and 2-senecioyl-4-vinylphenol (8.5-41.0%) were the most prominent constituents. The former two compounds were isolated and characterized by spectroscopic data. The essential oils and the isolated compounds were tested against Aedes aegypti larvae survival. The results obtained show that the essential oil from roots and 2,2-dimethyl-6-vinylchroman-4-one (10.3-25.5%) could be considered as natural larvicidal agents.

Paracoccidioidomycosis is a systemic mycosis endemic in Latin America, with a high prevalence in Brazil, Argentina, Colombia and Venezuela. The aetiological agent of disease is the thermal dimorphic fungus, Paracoccidioides brasiliensis. A glycoprotein of 43 kD (gp43) is the major antigen of P. brasiliensis. Antibodies directed to this antigen are detected in the sera of all patients with paracoccidioidomycosis (PCM). Recently, it has been shown that mice immunized with anti-gp43 monoclonal antibodies (MAbs) (Ab1), induce the idiotypic cascade in the gp43 system, which produced both, anti-Id antibodies (Ab2) and anti-anti-Id antibodies (Ab3). To further characterize the idiotypic cascade modulation in mice immunized with anti-gp43 MAb 17c, hybridomas were produced. Ab2 MAbs named 7.B12 inhibited (>95%) the binding of gp43 to MAb 17c (Ab1), suggesting that this anti-Id MAb bind to the idiotope, thus fulfilling the internal image criteria. To elucidate whether Ab2 MAb could act as antigen in serological assays, instead of gp43, sera from PCM patients were tested. Using an ELISA test, it was observed that antibodies from patients and not normal serum bound to Ab2. However, the ELISA test using Ab2 bound to the solid phase made possible to serologically monitor the patients after antifungal therapy, showing an equivalent curve when compared with ELISA test employing purified gp43. Our results also showed that, when mice were immunized with Ab2beta and their cells were exposed to gp43 in vitro, a T cell proliferation response was observed.

Intracerebroventricular (i.c.v.) administration of corticotropin-releasing factor (CRF) biphasically affects performance in tests of learning and memory. In the present study, we used CRF, urocortin (Ucn), a recently cloned CRF homologue, and CRF receptor antagonists, to determine which CRF receptor subtype(s) mediate the memory modulating effects of CRF receptor agonists in male Wistar rats. Under difficult learning conditions (massed trials), i.c.v. pretreatment with CRF or Ucn facilitated the acquisition of spatial navigation in the Morris water maze in a non-dose-dependent fashion (optimal doses of 0.1 and 0.03 microg, respectively). Under less difficult learning conditions (spaced trials), both peptides impaired water maze performance. In addition, with i.c.v. posttraining treatment, the peptides were equipotent (1.0 microg) in facilitating the consolidation of passive avoidance learning. The performance-enhancing effects of Ucn in both water maze and passive avoidance paradigms were reversed by i.c.v. pretreatment with D-Phe CRF(12-41) (2.5, 5 microg), a broad CRF(1)/CRF(2) receptor antagonist, or antalarmin (10 microg), a potent, nonpeptide, CRF(1) selective receptor antagonist. Thus, Ucn shares CRF's memory-modulating effects, and these effects appear to be mediated via the CRF(1) receptor. These findings are consistent with the hypothesis that CRF receptor agonists affect performance in tests of learning and memory by increasing arousal.

The present studies were designed to evaluate the competitive binding properties and functional effects of a novel nonpeptide CRF1 receptor antagonist, R121919. R121919 administered in doses of 0.63 to 20 mg/kg p.o. 60 min pretest in Wistar rats dose dependently attenuated the swim stress-induced anxiogenic-like behavior in the elevated plus-maze model of anxiety. Moreover, receptor autoradiography revealed that R121919 dose-dependently occupied brain CRF1 receptors in subjects tested in the plus-maze experiment. Orally administered doses of up to 20 mg/kg R121919 also blunted basal and swim stress-induced pituitary-adrenocortical activation, produced additional anxiolytic-like behavioral actions in the defensive withdrawal and defensive burying paradigms, and functionally antagonized the locomotor stimulatory properties of exogenously administered CRF. Taken together, these results suggest that the anxiolytic-like efficacy of R121919 in attenuating the stress-, novelty-, shock-, and CRF-induced increases in behavioral arousal is correlated with competitive blockade of central CRF1 receptors.

Panels of monoclonal antibodies (mAbs) were raised against recombinant human leptin and the recombinant human soluble leptin receptor. Using these mAbs, we established a ligand-mediated immunofunctional assay (LIFA) to quantify concentrations of the soluble leptin receptor, which has been shown to be a major binding protein for leptin in human serum. In performing the assay, a monoclonal antibody (mAb 2H6) against the soluble leptin receptor, which binds an epitope outside the leptin-binding site and equally recognizes both, free and leptin-occupied soluble leptin receptor, is used to capture the soluble leptin receptor on a microtiter plate. Recombinant human leptin is added to saturate all binding sites, and a biotinylated anti-leptin mAb (4D3) detects the amount of leptin (endogenous and exogenous) bound to the soluble leptin receptor. The same procedure, but without adding exogenous leptin, allows for measurement of the circulating endogenous leptin/soluble leptin receptor complexes. The LIFA assay has a linear working range of 0.5-200 microg/liter, intra- and interassay coefficients of variation ranged from 3.2-6.3% and from 5.2-7.9%, respectively. The assay has a linearity of 102.2 +/- 5.2% (mean +/- SD) and a recovery of 100.7 +/- 6.9%. Size-exclusion chromatography revealed that the assay measures a protein with a main peak eluted at 340 kDa. The soluble leptin receptor concentration (63.3 +/- 22.8 microg/liter (mean +/- SD), range 17.9-129.2 microg/liter, n = 43) in normal subjects (body mass index = 22.3 +/- 2.3 kg/m(2)) was not different from the concentration (54.4 +/- 19.8 microg/liter, range 23.7-104.8 microg/liter, n = 34, P > 0.05) found in obese subjects (body mass index = 40.9 +/- 15.7 kg/m(2)). However, the percentage of the total soluble leptin receptor complexed with endogenous leptin was significantly higher in obese subjects, compared with normal subjects (74.9% +/- 23.5% vs. 33.1% +/- 19.5%, P < 0.001). Higher serum leptin levels in obese subjects (38.4 +/- 23.7 microg/liter vs. 7.8 +/- 5.5 microg/liter in normal subjects, P < 0.001) together with comparable soluble leptin receptor levels result in a lower proportion of leptin bound to the soluble leptin receptor in obese subjects (19.3% +/- 19.4%, range 4.9-97.2%) than in normal subjects (39.0% +/- 22.5%, range 15.3-96.5%, P < 0.001). The development of this LIFA for the rapid and accurate quantification of total soluble leptin receptor and circulating leptin/soluble leptin receptor complexes provides a valuable tool for the further understanding of the role of leptin and its soluble receptor in health and disease.

Previous studies demonstrated that ether-laparotomy significantly increased iodine-125-labeled interleukin-1alpha ([125I]IL-1alpha) binding in the mouse anterior pituitary at 2 h after the onset of stress. Corticotropin-releasing factor (CRF) receptor antagonist, D-Phe CRF (12-41), abolished ether-laparotomy-induced increase in [125I]IL-1alpha binding in the pituitary, showing that CRF plays a pivotal role in the regulation of IL-1 receptors under stress conditions. In an attempt to define the effect of CRA 1000 (2-(N-(2-methylthio-4-isopropylphenyl)-N-ethylamino-4-(4-(3-fluorophenyl)-1,2,3,6-tetrahydropyridin-1-yl)-6-methylpyrimidine), a non-peptide CRF receptor type 1 antagonist on the regulation of hypothalamic-pituitary-adrenal (HPA) axis and IL-1 receptors in the mouse, we measured plasma adrenocorticotropic hormone (ACTH) and corticosterone levels, [125I]IL-1alpha binding and the expression of transcripts for type 1 IL-1 receptor (IL-1R1 mRNA) in the pituitary at 2 h after endotoxin lipopolysaccharide (LPS) treatment or ether-laparotomy stress with or without CRA 1000 pretreatment. A single injection of LPS dramatically increased plasma ACTH and corticosterone levels compared with saline injection. In contrast, plasma ACTH levels were significantly attenuated in response to one LPS injection following oral CRA 1000 pretreatment. LPS-induced plasma corticosterone levels tended to be lower after CRA 1000 pretreatment but it did not reach statistical significance. Ether-laparotomy stress significantly increased plasma ACTH and corticosterone levels at 2 h after the onset of stress and CRA 1000 pretreatment did not affect the peak ACTH and corticosterone levels following stress. Ether-laparotomy stress resulted in a robust increase in [125I]IL-1alpha binding and IL-1R1 mRNA levels in the pituitary. CRA 1000 pretreatment significantly decreased ether-laparotomy stress-induced IL-1R1 mRNA levels but did not affect [125I]IL-1alpha binding. Pretreatment with CRA 1000 without stress significantly increased [125I]IL-1alpha binding and IL-1R1 mRNA levels compared with those in vehicle pretreatment. These data demonstrate differential effects of CRA 1000 in HPA axis following endotoxin and ether-laparotomy stress and complex interactions between CRF and IL-1 receptors during stress.

Intracerebroventricular (i.c.v.) administration of corticotropin-releasing factor (CRF) peptide fragments with low affinity for CRF receptors reportedly improves cognitive performance without producing anxiety. These compounds are hypothesized to act by displacing endogenous peptide from the CRF-binding protein (CRF-BP). To test this hypothesis, the present study determined whether the performance-enhancing potency of CRF fragments was related to their affinity for the CRF-BP. Rank ordering of the optimal doses of these compounds for facilitating spatial navigation corresponded to their affinity for the CRF-BP. i.c.v. pretreatment with performance-enhancing doses of r/h CRF(1-41)-OH (5 micrograms) or r/h CRF(6-33) (25 micrograms) did not increase emotionality. These findings replicate the dissociability of the cognition- and anxiety-related effects of CRF-related compounds and suggest that CRF fragments facilitate performance via the CRF-BP.

Objective: To compare the effect of orchidectomy (ODX) in 7- and 24-week-old C57BL/6 mice on the age-related responses of the cytokine interleukin (IL)-1beta and its receptor to intraperitoneal injection of the bacterial endotoxin, lipopolysaccharide (LPS)., Methods: We measured IL-1beta concentrations in the plasma, hippocampus, hypothalamus and adrenal gland using ELISA and iodine-125-labeled recombinant human IL-1alpha ([(125)I]IL-1alpha) binding in the hippocampus following the intraperitoneal administration of saline or LPS., Results: There were no significant differences in the concentrations of IL-1beta and its receptors in the brain and peripheral tissues between sham-operated and ODX mice in both age groups injected with saline. LPS induced significantly higher IL-1beta production in the plasma and hippocampus in sham-operated 24-week-old mice than in 7-week-old mice. Coincident with the heightened IL-1beta response to LPS, hippocampal [(125)I]IL-1alpha binding was lower in 24-week-old mice than in 7-week-old mice after LPS injection in the sham-operated group. The age-related differences in the IL-1beta concentrations in the plasma and hippocampus and [(125)I]IL-1alpha binding in the hippocampus in response to LPS administration were abolished by ODX. Although LPS dramatically increased IL-1beta levels in the hypothalamus, no significant age-related differences in IL-1beta concentrations were seen, and ODX did not affect IL-1beta levels. In contrast, there were no significant differences between saline- and LPS-injected 7-week-old mice in relation to concentrations in the adrenal gland. Moreover, although the adrenal IL-1beta concentrations in 24-week-old mice were significantly higher than those in 7-week-old mice, ODX did not abolish these age-related differences in concentrations in the adrenal gland., Conclusions: Our data suggest the involvement of testosterone (or gonadal product) in plasma and hippocampal IL-1beta regulation in relation to age, and demonstrate the importance of the gonadal development in mediating the effects of infectious challenge on the brain and immune function., (Copyright 2000 S. Karger AG, Basel)