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1.
Appl Opt ; 60(1): 1-9, 2021 Jan 01.
Article in English | MEDLINE | ID: mdl-33362066

ABSTRACT

We show an in-line digital holographic image reconstruction from subsampled holograms with resolution improvement and lensless magnification with high noise immunity by a compressive sensing approach. Our method treats the sensed field as subsampled, low-pass filtered and projected on a Fresnel-Bluestein base in an inverse problem approach to image reconstruction with controlled lensless magnification. So, we have demonstrated by simulation and experimental results that the approach can reconstruct images with quality even when used in holograms obtained from unusual subsampling schemes.

2.
Epidemiol Infect ; 148: e126, 2020 06 22.
Article in English | MEDLINE | ID: mdl-32624035

ABSTRACT

Spontaneous abortion is considered a public health problem having several causes, including infections. Among the infectious agents, bacteria of the vaginal microbiota and Ureaplasma parvum have been associated with abortion, but their participation needs to be further elucidated. This study aims to evaluate the influence of Mollicutes on the development of spontaneous abortion. Women who underwent spontaneous abortion and those with normal birth (control) were studied. Samples of cervical mucus (CM) and placental tissue were collected to identify Mollicutes using the quantitative polymerase chain reaction methodology. Eighty-nine women who had a miscarriage and 20 women with normal pregnancies were studied. The presence of Mollicutes in placental tissue increased the chance of developing miscarriage sevenfold. The prevalence of U. parvum in women who experienced spontaneous abortion was 66.3% in placental tissue. A positive association was observed between the detection of U. parvum in samples of placental tissue and abortion. There was a significant increase in microbial load in placental tissue for M. hominis, U. urealyticum and U. parvum compared to the control group. Detection of U. parvum in CM in pregnant women can ascend to the region of the placental tissue and trigger a spontaneous abortion.


Subject(s)
Abortion, Spontaneous/microbiology , Ureaplasma Infections/microbiology , Ureaplasma/isolation & purification , Adult , Case-Control Studies , Cross-Sectional Studies , Female , Humans , Pregnancy , Risk Factors , Young Adult
3.
Scand J Med Sci Sports ; 27(7): 762-769, 2017 Jul.
Article in English | MEDLINE | ID: mdl-27230405

ABSTRACT

This study compared the physiological strain induced by prolonged walking and running performed at the walk-run transition speed (WRTS) in healthy untrained men. Twenty volunteers (age: 28 ± 5.01 years; height: 174.0 ± 0.3 cm; body mass: 74.5 ± 0.6 kg) underwent the following: (a) ramp-incremental maximal cardiopulmonary exercise test (CPET); (b) specific protocol to detect the WRTS; and (c) two 30-min walking and running bouts at WRTS (mean ± SD: 6.9 ± 0.06 km/h). Expired gases were collected during exercise bouts via the metabolic cart. A significant effect of locomotion mode (F = 4.8, P < 0.001) was observed with running resulting in higher cardiorespiratory responses than walking at the WRTS (oxygen uptake: mean difference = 0.26 L/min; pulmonary ventilation: mean difference = 5.53 L/min; carbon dioxide output: mean difference = 0.32 L/min; heart rate: mean difference = 13 beats/min; total energy expenditure: mean difference = 59 kcal). The rating of perceived exertion was similar across locomotion modes (mean difference = 0.3; P = 0.490). In conclusion, running promoted greater cardiorespiratory responses than walking at the WRTS in untrained healthy men. These data might have practical impact on aerobic training performed at intensities corresponding to WRTS.


Subject(s)
Exercise Test , Physical Exertion/physiology , Running/physiology , Walking/physiology , Adult , Heart Rate , Humans , Male , Oxygen Consumption , Pulmonary Ventilation , Young Adult
4.
Int J Obes (Lond) ; 40(3): 479-86, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26443339

ABSTRACT

BACKGROUND/OBJECTIVES: The association between gluten and body weight is inconsistent. Previously, we showed that a gluten-free diet reduces weight gain without changing food intake in mice fed high-fat diets. In the present study, we investigated the effects of gluten intake on fat metabolism, thermogenesis and energy expenditure in mice fed a standard or high-fat diet. METHODS: Mice were fed four different experimental diets during 8 weeks: a control-standard diet (CD), a CD added with 4.5% of wheat gluten (CD-G), a high-fat diet (HFD) and a HFD added with 4.5% of wheat gluten (HFD-G). After 8 weeks, the mice received (99m)Tc-radiolabeled gluten orally to study gluten absorption and biodistribution or they underwent indirect calorimetry. After killing, subcutaneous and brown adipose tissues (SAT and BAT) were collected to assess thermogenesis-related protein expression. Lipid metabolism was studied in adipocyte cultures from the four groups. RESULTS: Despite having had the same energy intake, CD-G and HFD-G mice exhibited increased body weight and fat deposits compared with their respective controls. (99m)Tc-GLU or its peptides were detected in the blood, liver and visceral adipose tissue, suggesting that gluten can even reach extraintestinal organs. Uncoupling protein-1 expression was reduced in the BAT of HFD-G and in the SAT of CD-G and HFD-G mice. Indirect calorimetry showed lower oxygen volume consumption in CD-G and HFD-G groups compared with their controls. In HFD mice, daily energy expenditure was reduced with gluten intake. Gluten also reduced adiponectin, peroxisome proliferator-activated receptor (PPAR)-α and PPARγ and hormone-sensitive lipase in cultures of isolated adipocytes from HFD mice, whereas in the CD-G group, gluten intake increased interleukin-6 expression and tended to increase that of tumor necrosis factor. CONCLUSIONS: Wheat gluten promotes weight gain in animals on both HFD and CD, partly by reducing the thermogenic capacity of adipose tissues.


Subject(s)
Energy Metabolism/physiology , Glutens , Obesity/metabolism , Weight Gain/physiology , Adipogenesis , Adiposity , Animals , Disease Models, Animal , Energy Intake , Feeding Behavior , Gene Expression Regulation , Lipid Metabolism , Male , Mice , Mice, Inbred C57BL , Thermogenesis
5.
Heliyon ; 10(10): e30623, 2024 May 30.
Article in English | MEDLINE | ID: mdl-38770291

ABSTRACT

The Hertz-Sneddon elastic indentation model is widely adopted in the biomechanical investigation of living cells and other soft materials using atomic force microscopy despite the explicit viscoelastic nature of these materials. In this work, we demonstrate that an exact analytical viscoelastic force model for power-law materials, can be interpreted as a time-dependent Hertz-Sneddon-like model. Characterizing fibroblasts (L929) and osteoblasts (OFCOLII) demonstrates the model's accuracy. Our results show that the difference between Young's modulus EY obtained by fitting force curves with the Hertz-Sneddon model and the effective Young's modulus derived from the viscoelastic force model is less than 3%, even when cells are probed at large forces where nonlinear deformation effects become significant. We also propose a measurement protocol that involves probing samples at different indentation speeds and forces, enabling the construction of the average viscoelastic relaxation function of samples by conveniently fitting the force curves with the Hertz-Sneddon model.

6.
Adv Colloid Interface Sci ; 328: 103166, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38728773

ABSTRACT

Despite significant efforts by scientists in the development of advanced nanotechnology materials for smart diagnosis devices and drug delivery systems, the success of clinical trials remains largely elusive. In order to address this biomedical challenge, magnetic nanoparticles (MNPs) have gained attention as a promising candidate due to their theranostic properties, which allow the simultaneous treatment and diagnosis of a disease. Moreover, MNPs have advantageous characteristics such as a larger surface area, high surface-to-volume ratio, enhanced mobility, mass transference and, more notably, easy manipulation under external magnetic fields. Besides, certain magnetic particle types based on the magnetite (Fe3O4) phase have already been FDA-approved, demonstrating biocompatible and low toxicity. Typically, surface modification and/or functional group conjugation are required to prevent oxidation and particle aggregation. A wide range of inorganic and organic molecules have been utilized to coat the surface of MNPs, including surfactants, antibodies, synthetic and natural polymers, silica, metals, and various other substances. Furthermore, various strategies have been developed for the synthesis and surface functionalization of MNPs to enhance their colloidal stability, biocompatibility, good response to an external magnetic field, etc. Both uncoated MNPs and those coated with inorganic and organic compounds exhibit versatility, making them suitable for a range of applications such as drug delivery systems (DDS), magnetic hyperthermia, fluorescent biological labels, biodetection and magnetic resonance imaging (MRI). Thus, this review provides an update of recently published MNPs works, providing a current discussion regarding their strategies of synthesis and surface modifications, biomedical applications, and perspectives.


Subject(s)
Metal Nanoparticles , Surface Properties , Animals , Humans , Drug Delivery Systems , Magnetic Resonance Imaging , Magnetite Nanoparticles/chemistry , Metal Nanoparticles/chemistry
7.
Minerva Gastroenterol Dietol ; 57(4): 361-7, 2011 Dec.
Article in English | MEDLINE | ID: mdl-22105724

ABSTRACT

AIM: The occurrence of silent myocardial ischemia during endoscopic retrograde cholangiopancreatography (ERCP) has been documented, but its clinical significance remains unknown. The aims of this study were to investigate the incidence and risk factors of myocardial ischemia during ERCP, to determine the presence or absence of permanent myocardial injury and to evaluate if deep sedation with propofol had a positive effect on myocardial ischemia during ERCP. METHODS: Ambulatory ST-segment monitoring from 30 minutes prior to 4 hours after ERCP was obtained on 50 patients. A deep sedation was performed with intravenous propofol administered by anesthesiologist. Changes in vital signs during ERCP, pre and postprocedural 12-lead ECG examination and cardiac enzymes were evaluated. RESULTS: Silent cardiac ischemia occurred only in one patient (2%) during ERCP. This 64-year-old patient did not develop hypoxemia, tachycardia or hypotension periods during the exam. None of the patients developed cardiac enzymes or postprocedural electrocardiographic changes. Thirty seven (74%) patients suffered rhythm changes. CONCLUSION: Although rhythm disturbances were common, silent myocardial ischemia during ERCP was rare (2%) and without clinical relevance. In prolonged or complex therapeutic procedures, like ERCP, deep sedation with propofol performed by trained personnel is associated with reduced cardiac complications.


Subject(s)
Cholangiopancreatography, Endoscopic Retrograde/adverse effects , Intraoperative Complications/etiology , Myocardial Ischemia/etiology , Adult , Aged , Aged, 80 and over , Deep Sedation , Female , Humans , Incidence , Intraoperative Complications/epidemiology , Intraoperative Complications/prevention & control , Male , Middle Aged , Myocardial Ischemia/epidemiology , Myocardial Ischemia/prevention & control , Prospective Studies , Risk Factors , Young Adult
8.
J Infect Dis ; 202(11): 1708-12, 2010 Dec 01.
Article in English | MEDLINE | ID: mdl-20977342

ABSTRACT

The increased incidence and severity of Clostridium difficile infection (CDI) in older adults (age, ≥65 years) corresponds with the emergence of the BI/NAP1 strain, making elucidation of the host immune response extremely important. We therefore infected germ-free C57BL/6 mice aged 7-14 months with a BI/NAP1 strain and monitored the mice for response. Infected mice were moribund 48-72 h after infection and developed gross and histological cecitis and colitis and elevated concentrations of keratinocyte chemoattractant, interleukin 1ß, monocyte chemotactic protein 1, and granulocyte colony-stimulating factor and decreased levels of interferon γ, interleukin 12 p40, interleukin 12 p70, and interleukin 10 compared with controls. We conclude that aged, germ-free C57BL/6 mice are susceptible to fulminant CDI from a BI/NAP1 strain and represent a novel model to further elucidate the host immune response to acute CDI.


Subject(s)
Clostridioides difficile/immunology , Clostridioides difficile/pathogenicity , Enterocolitis, Pseudomembranous/microbiology , Animals , Clostridioides difficile/classification , Colon/microbiology , Colon/pathology , Disease Models, Animal , Enterocolitis, Pseudomembranous/immunology , Enterocolitis, Pseudomembranous/pathology , Germ-Free Life , Granulocyte Colony-Stimulating Factor/analysis , Interferon-gamma/analysis , Interleukin-10/analysis , Interleukin-12/analysis , Interleukin-12 Receptor beta 1 Subunit/analysis , Interleukin-1beta/analysis , Mice , Mice, Inbred C57BL
9.
Sci Rep ; 10(1): 4749, 2020 03 16.
Article in English | MEDLINE | ID: mdl-32179816

ABSTRACT

Living cells are constantly exchanging momentum with their surroundings. So far, there is no consensus regarding how cells respond to such external stimuli, although it reveals much about their internal structures, motility as well as the emergence of disorders. Here, we report that twelve cell lines, ranging from healthy fibroblasts to cancer cells, hold a ubiquitous double power-law viscoelastic relaxation compatible with the fractional Kelvin-Voigt viscoelastic model. Atomic Force Microscopy measurements in time domain were employed to determine the mechanical parameters, namely, the fast and slow relaxation exponents, the crossover timescale between power law regimes, and the cell stiffness. These cell-dependent quantities show strong correlation with their collective migration and invasiveness properties. Beyond that, the crossover timescale sets the fastest timescale for cells to perform their biological functions.


Subject(s)
Cell Physiological Phenomena/physiology , Elasticity , Viscosity , Cell Line , Cell Line, Tumor , Cell Movement , Fibroblasts/physiology , Humans , Microscopy, Atomic Force , Models, Biological , Molecular Imaging , Neoplasm Invasiveness/pathology
10.
Mem Inst Oswaldo Cruz ; 104(2): 378-82, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19430668

ABSTRACT

The aim of this study was to evaluate associations between seropositivity for IgG and IgM anti-Toxoplasma gondii antibodies and socio-economic and environmental variables in pregnant women of Londrina, state of Paraná, Brazil. We interviewed 492 pregnant women, each of whom answered an epidemiological questionnaire, and collected blood samples for measurement of IgG and IgM anti-T. gondii antibodies by chemiluminescence. A confirmatory diagnosis of acute infection was made by an IgG avidity test. Titres of specific IgG anti-T. gondii were obtained by IFAT. Seropositivity for IgG anti-T. gondii antibodies was observed in 242 women (49.2%) and, of these, six pregnant women (1.2%) showed seropositivity for IgM. Age group, level of education, per capita income, presence of a cat in the house and a habit of eating green vegetables were all factors associated with a greater chance of infection with T. gondii. This study showed that 250 (50.8%) pregnant women were susceptible to T. gondii and considered to be at high risk for toxoplasmosis during pregnancy. Based on the results obtained, is critical to establish a program of health surveillance for toxoplasmosis, in order to contribute to diagnosis and early treatment during the prenatal period. It is also necessary to introduce measures to prevent the Toxoplasma infection in seronegative pregnant women.


Subject(s)
Antibodies, Protozoan/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Pregnancy Complications, Parasitic/epidemiology , Toxoplasmosis/epidemiology , Adolescent , Adult , Animals , Brazil/epidemiology , Cats , Female , Humans , Pregnancy , Pregnancy Complications, Parasitic/diagnosis , Pregnancy Trimester, First , Prevalence , Risk Factors , Socioeconomic Factors , Toxoplasmosis/diagnosis , Young Adult
11.
Equine Vet J ; 41(5): 487-92, 2009 May.
Article in English | MEDLINE | ID: mdl-19642410

ABSTRACT

REASONS FOR PERFORMING STUDY: It is unknown if different locomotor activities are equally effective at meeting the stabled horse's need for exercise and if they attenuate unwanted behaviour. HYPOTHESIS: Alternative forms of exercise influence the intensity of locomotor activities during a period of turn-out (the so-called rebound effect) and the occurrence of unwanted or undesirable activities during standard handling situations. METHOD: Twenty-four horses kept in stables were randomly assigned to one of 4 exercise regimes (walker, treadmill, turn-out and riding) for 4 consecutive days. Because these forms of exercise provide additional environmental stimulation, beyond that provided by exercise, each horse served as its own control in 4 corresponding (no exercise) control treatments presented in a balanced order. Unwanted behaviour was tested by taking horses to weighing scales and loading and unloading them onto a 4-horse float by an experienced handler and the rebound effect was tested by releasing them into a large arena for a period of 15 min at the end of the exercise and control treatments. RESULTS: Locomotor activities made up a large part of behaviour in the large arena following control treatments and all exercise regimes were sufficient to reduce the intensity of walking (P < 0.05), trotting (P < 0.01) and cantering (P < 0.001) on release into a large arena. Exercise regime reduced the number of bucks (P < 0.01) and rolling (P < 0.05) during rebound tests suggesting that turn-out was having a stronger effect than the other 3 exercise regimes. Exercise regimes significantly reduced the amount of unwanted behaviour and the number of commands given by the handler during weighing (P < 0.05) but had no effect on these behaviours during loading onto a float. CONCLUSION: Providing stabled horses with one hour/day of exercise on a walker, treadmill, turn-out or by being ridden are all effective at allowing expression of locomotor activities in stabled horses. POTENTIAL RELEVANCE: Providing stabled horses with regular exercise is likely to provide positive effects on horse welfare, training ability and handler safety.


Subject(s)
Behavior, Animal/physiology , Horses/physiology , Physical Conditioning, Animal/physiology , Animal Husbandry , Animals , Housing, Animal , Motor Activity/physiology
12.
Rev Gastroenterol Mex (Engl Ed) ; 84(3): 284-289, 2019.
Article in English, Spanish | MEDLINE | ID: mdl-30107945

ABSTRACT

INTRODUCTION AND AIMS: Interval colorectal cancer (iCRC) can occur due to missed lesions or to a newly developed lesion. The present study aimed to assess the iCRC rate and its characteristics in our population and find possible explanations. MATERIALS AND METHODS: A retrospective study was conducted on patients with colorectal cancer (CRC) diagnosed between January 2011 and January 2015 at our department. Demographics, endoscopic data, and tumor characteristics (location, histology, staging) were collected. We identified patients diagnosed with CCR who underwent colonoscopy at our department in the previous 10years and presented the disease (iCRC) before the date of their next recommended exam. The cases of iCRC were characterized and compared with other CRC cases. Possible explanations for the appearance of iCRC were analyzed. RESULTS: A total of 266 patients presented with CRC, 61.7% were men, and mean patient age was 70.7years. We identified 10 patients with iCRC: 6 were men, and mean patient age was 71.1years. Mean time for iCRC diagnosis after index colonoscopy was 3.5±1.84years. Tumor was located in the right colon in 50% of the patients with iCRC and in 24.5% of the patients without iCRC (P=.091). More patients with iCRC had a family history of CRC (50%) than the patients with reference CRC (3.1%) (P=.000). CONCLUSIONS: In our case series, 3.76% of all CRC were iCRC. There were no statistically significant differences between patients with or without iCRC, with the exception of family history of CRC.


Subject(s)
Adenocarcinoma/diagnosis , Colonoscopy/methods , Colorectal Neoplasms/diagnosis , Adenocarcinoma/epidemiology , Adenocarcinoma/surgery , Adult , Aged , Aged, 80 and over , Colorectal Neoplasms/epidemiology , Colorectal Neoplasms/surgery , Early Detection of Cancer , Female , Humans , Male , Middle Aged , Prevalence , Retrospective Studies , Treatment Outcome
13.
Oncogene ; 38(24): 4886, 2019 Jun.
Article in English | MEDLINE | ID: mdl-31068665

ABSTRACT

The final sentence of the Acknowledgements should be as follows: This work was supported by grants from Instituto de Salud Carlos III (BA15/00092), Spanish Ministry of Economy and Competitiveness/EU-ERDF (SAF2016-80626-R, SAF2013-49149-R, BFU2014-51672-REDC), Fundación CajaCanarias (AP2015/008) to RF, and the Australian National Health and Medical Research (NHMRC program grant to SRL and KKK (APP1017028).

14.
Cell Death Differ ; 14(8): 1433-42, 2007 Aug.
Article in English | MEDLINE | ID: mdl-17431426

ABSTRACT

Apoptosis plays a crucial role in development and tissue homeostasis. Some key survival pathways, such as DNA damage checkpoints and DNA repair, have been described to be inactivated during apoptosis. Here, we describe the processing of the human checkpoint protein Claspin during apoptosis. We observed cleavage of Claspin into multiple fragments in vivo. In vitro cleavage with caspases 3 and 7 of various fragments of the protein, revealed cut sites near the N- and C-termini of the protein. Using mass spectrometry, we identified a novel caspase cleavage site in Claspin at Asp25. Importantly, in addition to cleavage by caspases, we observed a proteasome-dependent degradation of Claspin under apoptotic conditions, resulting in a reduction of the levels of both full-length Claspin and its cleavage products. This degradation was not dependent upon the DSGxxS phosphodegron motif required for SCF(beta-TrCP)-mediated ubiquitination of Claspin. Finally, downregulation of Claspin protein levels by short interfering RNA resulted in an increase in apoptotic induction both in the presence and absence of DNA damage. We conclude that Claspin has antiapoptotic activity and is degraded by two different pathways during apoptosis. The resulting disappearance of Claspin from the cells further promotes apoptosis.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Apoptosis/physiology , Caspases/metabolism , Proteasome Endopeptidase Complex/metabolism , Adaptor Proteins, Signal Transducing/antagonists & inhibitors , Adaptor Proteins, Signal Transducing/chemistry , Adaptor Proteins, Signal Transducing/genetics , Amino Acid Motifs , Ataxia Telangiectasia Mutated Proteins , Base Sequence , Binding Sites , Caspase 3/metabolism , Caspase 7/metabolism , Cell Cycle Proteins/metabolism , Checkpoint Kinase 1 , DNA Damage , HL-60 Cells , HeLa Cells , Humans , In Vitro Techniques , Models, Biological , Peptide Fragments/chemistry , Peptide Fragments/genetics , Peptide Fragments/metabolism , Protein Kinases/metabolism , Protein Serine-Threonine Kinases/metabolism , RNA, Small Interfering/genetics , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism
15.
Vet Rec ; 163(14): 413-7, 2008 Oct 04.
Article in English | MEDLINE | ID: mdl-18836155

ABSTRACT

The effects of dietary supplements of virginiamycin on the behaviour and physiology of 17 thoroughbred geldings (five cribbers, six weavers and six control horses) were compared with the effects of a placebo over a period of 16 weeks. Virginiamycin had no effect on the horses' stereotypic behaviour, but it reduced their explorative behaviour, possibly owing to a reduction in feeding motivation. Virginiamycin increased the water intake of the cribbers and decreased the water intake of the control horses, but it was not possible to eliminate possible confounding factors for this effect. Virginiamycin had no other significant effects on the behaviour or physiology of the horses, and had no effect on the digestibility of their diets.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Horses , Stereotyped Behavior/drug effects , Virginiamycin/therapeutic use , Animals , Drinking , Male
16.
Ultrason Sonochem ; 48: 340-348, 2018 Nov.
Article in English | MEDLINE | ID: mdl-30080559

ABSTRACT

TiO2 is a common inorganic filter used in sunscreens due to its photoprotective effect on the skin against UV radiation. However, the use of this kind of material in cosmetics is limited by its inherent photocatalytic activity. It is known that coating on TiO2 surface can improve some features. Although, many of the methodologies used for this purpose are still laborious and time-consuming. Thus, this work reports a novel, easy, cheap and fast strategy to coat TiO2 particles by using a sonochemistry approach, aiming to decrease photocatalytic activity and to enhance colloidal stability. For this proposal, SiO2, Al2O3, ZrO2 and sodium polyacrylate (PAANa) were used to tune the surface of commercial TiO2 particles and they were applied in a sunscreen formulation. The samples were characterized by XRPD, FT-IR, DLS, EDS, SEM and TEM. The photocatalytic activity and UV-shielding ability were also evaluated. The sunscreen formulations were prepared and characterized by zeta potential, DLS, and Sun Protection Factor (SPF). FT-IR, EDS, and charge surface of the particles confirmed the success of the sonochemistry coating. Additionally, TiO2@Al2O3, TiO2@SiO2 and TiO2@PAANa show a lower photocatalytic activity than original TiO2 with similar UV-shielding ability. The sunscreens produced with the coated TiO2 have similar SPF to the one with commercial TiO2. Specifically, the sunscreen with TiO2@PAANa shows an increase in colloidal stability. Herein, the incorporation of the sonochemical-coated TiO2 particles in sunscreen formulations may produce sunscreens with better aesthetic appearance and a greater health security due to its lower free radicals production.

17.
Res Vet Sci ; 82(2): 202-7, 2007 Apr.
Article in English | MEDLINE | ID: mdl-17266999

ABSTRACT

Sheep and dog blood samples were collected from nine farms in the county of Guarapuava, Paraná, Brazil. The indirect fluorescent antibody test (IFAT) was used to detect Neospora caninum and Toxoplasma gondii antibodies. Herein, serum samples from 305 sheep were evaluated, being 29 (9.5%) and 157 (51.5%) seropositives to N. caninum and T. gondii, respectively. Seven (29.1%) and five (20.8%) out of 24 dogs were seropositives to N. caninum and T. gondii, respectively. There were no differences among the sheep serology for N. caninum and reproductive problems, management and animal feeding variables, neurological problems and presence of other animals species on the farm (P>or=0.05). The simultaneous frequency of antibodies between N. caninum and T. gondii was 5.2% in the herds. Age, breed, farm size, semi-intensive activity, mineral salt supplementation, water origin, stage of the pregnancy when reproduction problems occurred, neurological problems in lambs, presence of rodents in the food room and pasture cat access were identified as associated factors for the occurrence of toxoplasmosis in sheep (P<0.05). There were no differences among the seropositivity in dogs for N. caninum and T. gondii and breed, age and sex (P>or=0.05). The present work is the first report on serum prevalence of N. caninum in sheep from the state of Paraná, Brazil.


Subject(s)
Coccidiosis/epidemiology , Coccidiosis/veterinary , Dog Diseases/parasitology , Neospora/isolation & purification , Sheep Diseases/parasitology , Toxoplasma/isolation & purification , Toxoplasmosis, Animal/epidemiology , Animal Husbandry , Animals , Antibodies, Protozoan/blood , Brazil/epidemiology , Coccidiosis/parasitology , Coccidiosis/transmission , Dog Diseases/epidemiology , Dog Diseases/transmission , Dogs , Female , Fluorescent Antibody Technique, Indirect/veterinary , Male , Pregnancy , Seroepidemiologic Studies , Sheep , Sheep Diseases/epidemiology , Sheep Diseases/transmission , Toxoplasmosis, Animal/parasitology , Toxoplasmosis, Animal/transmission
18.
Oncogene ; 36(33): 4802-4809, 2017 08 17.
Article in English | MEDLINE | ID: mdl-28288134

ABSTRACT

Correct control of DNA replication is crucial to maintain genomic stability in dividing cells. Inappropriate re-licensing of replicated origins is associated with chromosomal instability (CIN), a hallmark of cancer progression that at the same time provides potential opportunities for therapeutic intervention. Geminin is a critical inhibitor of the DNA replication licensing factor Cdt1. To properly achieve its functions, Geminin levels are tightly regulated through the cell cycle by ubiquitin-dependent proteasomal degradation, but the de-ubiquitinating enzymes (DUBs) involved had not been identified. Here we report that DUB3 and USP7 control human Geminin. Overexpression of either DUB3 or USP7 increases Geminin levels through reduced ubiquitination. Conversely, depletion of DUB3 or USP7 reduces Geminin levels, and DUB3 knockdown increases re-replication events, analogous to the effect of Geminin depletion. In exploring potential clinical implications, we found that USP7 and Geminin are strongly correlated in a cohort of invasive breast cancers (P<1.01E-08). As expected, Geminin expression is highly prognostic. Interestingly, we found a non-monotonic relationship between USP7 and breast cancer-specific survival, with both very low or high levels of USP7 associated with poor outcome, independent of estrogen receptor status. Altogether, our data identify DUB3 and USP7 as factors that regulate DNA replication by controlling Geminin protein stability, and suggest that USP7 may be involved in Geminin dysregulation during breast cancer progression.


Subject(s)
Breast Neoplasms/enzymology , Cell Cycle Proteins/antagonists & inhibitors , Endopeptidases/metabolism , Geminin/metabolism , Ubiquitin Thiolesterase/metabolism , Ubiquitin/metabolism , Breast Neoplasms/diagnosis , Breast Neoplasms/pathology , Cell Cycle , Cell Line, Tumor , Chromosomal Instability , DNA Replication/physiology , Disease Progression , Endopeptidases/genetics , HEK293 Cells , Humans , Kaplan-Meier Estimate , Neoplasm Invasiveness , Prognosis , Protein Stability , RNA, Small Interfering/genetics , Ubiquitin Thiolesterase/genetics , Ubiquitin-Specific Peptidase 7 , Ubiquitination
19.
Oncogene ; 36(5): 678-686, 2017 02 02.
Article in English | MEDLINE | ID: mdl-27375025

ABSTRACT

Stresses such as hypoxia, nutrient deprivation and acidification disturb protein folding in the endoplasmic reticulum (ER) and activate the Unfolded Protein Response (UPR) to trigger adaptive responses through the effectors, PERK, IRE1 and ATF6. Most of these responses relate to ER homoeostasis; however, here we show that the PERK branch of the UPR also controls DNA replication. Treatment of cells with the non-genotoxic UPR agonist thapsigargin led to a rapid inhibition of DNA synthesis that was attributable to a combination of DNA replication fork slowing and reduced replication origin firing. DNA synthesis inhibition was dependent on the UPR effector PERK and was associated with phosphorylation of the checkpoint adaptor protein Claspin and activation of the Chk1 effector kinase, both of which occurred in the absence of detectable DNA damage. Remarkably, thapsigargin did not inhibit bulk DNA synthesis or activate Chk1 in cells depleted of Claspin, or when Chk1 was depleted or subject to chemical inhibition. In each case thapsigargin-resistant DNA synthesis was due to an increase in replication origin firing that compensated for reduced fork progression. Taken together, our results unveil a new aspect of PERK function and previously unknown roles for Claspin and Chk1 as negative regulators of DNA replication in the absence of genotoxic stress. Because tumour cells proliferate in suboptimal environments, and frequently show evidence of UPR activation, this pathway could modulate the response to DNA replication-targeted chemotherapies.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Checkpoint Kinase 1/metabolism , DNA Replication/physiology , Unfolded Protein Response/physiology , eIF-2 Kinase/metabolism , Adaptor Proteins, Signal Transducing/genetics , Cell Line, Tumor , Checkpoint Kinase 1/genetics , Humans , Transfection , eIF-2 Kinase/genetics
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