ABSTRACT
In the aftermath of the COVID-19 pandemic, the evolution of the SARS-CoV-2 into a seasonal pathogen along with the emergence of new variants, underscores the need for dynamic and adaptable responses, emphasizing the importance of sustained vaccination strategies. This observer-blind, double-dummy, randomized immunobridging phase 2 study (NCT05175742) aimed to compare the immunogenicity induced by two doses of 40 µg PTX-COVID19-B vaccine candidate administered 28 days apart, with the response induced by two doses of 30 µg Pfizer-BioNTech COVID-19 vaccine (BNT162b2), administered 21 days apart, in Nucleocapsid-protein seronegative adults 18-64 years of age. Both vaccines were administrated via intramuscular injection in the deltoid muscle. Two weeks after the second dose, the neutralizing antibody (NAb) geometric mean titer ratio and seroconversion rate met the non-inferiority criteria, successfully achieving the primary immunogenicity endpoints of the study. PTX-COVID19-B demonstrated similar safety and tolerability profile to BNT162b2 vaccine. The lowest NAb response was observed in subjects with low-to-undetectable NAb at baseline or no reported breakthrough infection. Conversely, participants who experienced breakthrough infections during the study exhibited higher NAb titers. This study also shows induction of cell-mediated immune (CMI) responses by PTX-COVID19-B. In conclusion, the vaccine candidate PTX-COVID19-B demonstrated favourable safety profile along with immunogenicity similar to the active comparator BNT162b2 vaccine.
Subject(s)
COVID-19 Vaccines , COVID-19 , Adult , Humans , Antibodies, Neutralizing , BNT162 Vaccine , CD59 Antigens , COVID-19/prevention & control , COVID-19 Vaccines/adverse effects , mRNA Vaccines , Pandemics , SARS-CoV-2ABSTRACT
J-domain proteins tune the specificity of Hsp70s, engaging them in precise functions. Despite their essential role, the structure and function of many J-domain proteins remain largely unknown. We explore human DNAJA2, finding that it reversibly forms highly-ordered, tubular structures that can be dissociated by Hsc70, the constitutively expressed Hsp70 isoform. Cryoelectron microscopy and mutational studies reveal that different domains are involved in self-association. Oligomer dissociation into dimers potentiates its interaction with unfolded client proteins. The J-domains are accessible to Hsc70 within the tubular structure. They allow binding of closely spaced Hsc70 molecules that could be transferred to the unfolded substrate for its cooperative remodelling, explaining the efficient recovery of DNAJA2-bound clients. The disordered C-terminal domain, comprising the last 52 residues, regulates its holding activity and productive interaction with Hsc70. These in vitro findings suggest that the association equilibrium of DNAJA2 could regulate its interaction with client proteins and Hsc70.
Subject(s)
HSP70 Heat-Shock Proteins , Polymers , Humans , Cryoelectron Microscopy , HSP40 Heat-Shock Proteins , MutationABSTRACT
Access to vaccines against SARS-CoV-2 virus was limited in poor countries during the COVID-19 pandemic. Therefore, a low-cost mRNA vaccine, PTX-COVID19-B, was produced and evaluated in a Phase 1 trial. PTX-COVID19-B encodes Spike protein D614G variant without the proline-proline (986-987) mutation present in other COVID-19 vaccines. The aim of the study was to evaluate safety, tolerability, and immunogenicity of PTX-COVID19-B vaccine in healthy seronegative adults 18-64 years old. The trial design was observer-blinded, randomized, placebo-controlled, and tested ascending doses of 16-µg, 40-µg, or 100-µg in a total of 60 subjects who received two intramuscular doses, 4 weeks apart. Participants were monitored for solicited and unsolicited adverse events after vaccination and were provided with a Diary Card and thermometer to report any reactogenicity during the trial. Blood samples were collected on baseline, days 8, 28, 42, 90, and 180 for serum analysis of total IgG anti-receptor binding domain (RBD)/Spike titers by ELISA, and neutralizing antibody titers by pseudovirus assay. Titers in BAU/mL were reported as geometric mean and 95% CI per cohort. After vaccination, few solicited adverse events were observed and were mild to moderate and self-resolved within 48 h. The most common solicited local and systemic adverse event was pain at the injection site, and headache, respectively. Seroconversion was observed in all vaccinated participants, who showed high antibody titers against RBD, Spike, and neutralizing activity against the Wuhan strain. Neutralizing antibody titers were also detected against Alpha, Beta, and Delta variants of concerns in a dose dependent manner. All tested doses of PTX-COVID19-B were safe, well-tolerated, and provided a strong immunogenicity response. The 40-µg dose showed fewer adverse reactions than the 100-µg dose, and therefore was selected for a Phase 2 trial, which is currently ongoing.Clinical Trial Registration number: NCT04765436 (21/02/2021). ( https://clinicaltrials.gov/ct2/show/NCT04765436 ).
Subject(s)
COVID-19 Vaccines , COVID-19 , Adult , Humans , Adolescent , Young Adult , Middle Aged , COVID-19 Vaccines/adverse effects , SARS-CoV-2/genetics , COVID-19/prevention & control , Pandemics/prevention & control , mRNA Vaccines , Antibodies, Neutralizing , Immunogenicity, Vaccine , Antibodies, Viral , Double-Blind MethodABSTRACT
Toxoplasma gondii is a significant threat to endangered Hawaiian wildlife including birds and marine mammals. To estimate the prevalence of T. gondii in stranded cetaceans from 1997 to 2021 in Hawai'i, we tested tissues from 37 stranded spinner dolphins Stenella longirostris and 51 stranded individuals that represented 18 other cetacean species. DNA from cetacean tissue extracts were screened using a nested polymerase chain reaction (PCR) assay targeting the Toxoplasmatinae internal transcribed spacer 1 of the nuclear ribosomal DNA. A positive result was obtained in 9 tissues examined for each of 2 spinner dolphins out of 525 tissue samples analyzed by PCR. The PCR-positive spinner dolphins had disseminated acute toxoplasmosis with necrosis, inflammation, and intralesional protozoal cysts and tachyzoites in multiple organs. Discrete positive immunostaining for T. gondii was observed in all tissues tested including the adrenal gland, brain, liver, and lung. Both positive spinner dolphins were negative for cetacean morbillivirus. The T. gondii genotyping was performed by restriction fragment length polymorphism (PCR-RFLP) based on 10 genetic markers. The PCR-RFLP analysis revealed the T. gondii belonged to PCR-RFLP-ToxoDB genotype #24, previously detected in wild pig Sus scrofa in O'ahu, bobcats Lynx rufus from Mississippi, USA, and chickens Gallus gallus from Costa Rica and Brazil. These cases represent the first report of this genotype in aquatic mammals and the second and third reports of fatal disseminated T. gondii infection in stranded spinner dolphins from Hawai'i. Nearshore species, like spinner dolphins, may be at increased risk of mortality from this parasite in marine coastal waterways via sewage systems, storm water drainage, and freshwater runoff.
Subject(s)
Stenella , Toxoplasma , Toxoplasmosis, Animal , Animals , Toxoplasma/genetics , Toxoplasmosis, Animal/epidemiology , Toxoplasmosis, Animal/parasitology , Hawaii/epidemiology , Prevalence , Chickens , Genotype , CetaceaABSTRACT
Consumption of undercooked meat is one of the main transmission routes for Toxoplasma gondii worldwide. In the South American Andes, the guinea pig (Cavia porcellus) is a domestic rodent representing one of the main sources of animal proteins for indigenous communities. Although T. gondii infects a wide range of rodents worldwide, the natural impact of the infection on guinea pig populations is still unknown. Our study conducted in guinea pigs that were bred in traditional systems located in the village of José María Hernández (Nariño, Colombia) revealed the presence of T. gondii antibodies in 33.3% (23 out of 69) guinea pigs evaluated, with a cut-off point of 25 for the modified direct agglutination test. Conventional PCR detection of the T. gondii-specific RE fragment (529 bp) in 207 collected tissues demonstrated the presence of T. gondii DNA in several organs, including the brain (16/69), muscle (12/69), and heart (4/69), with an overall molecular detection frequency of 27.5% (19 out of 69 guinea pigs). This is the first report of natural infection of guinea pigs with T. gondii, demonstrating their potential epidemiological role in transmitting the infection to autochthonous populations.
Subject(s)
Rodent Diseases , Swine Diseases , Toxoplasma , Toxoplasmosis, Animal , Swine , Animals , Guinea Pigs , Humans , Toxoplasma/genetics , Toxoplasmosis, Animal/diagnosis , Toxoplasmosis, Animal/epidemiology , Colombia/epidemiology , Swine Diseases/diagnosis , South America , RodentiaABSTRACT
Apg2, one of the three cytosolic Hsp110 chaperones in humans, supports reactivation of unordered and ordered protein aggregates by Hsc70 (HspA8). Together with DnaJB1, Apg2 serves to nucleate Hsc70 molecules into sites where productive entropic pulling forces can be developed. During aggregate reactivation, Apg2 performs as a specialized nucleotide exchange factor, but the origin of its specialization is poorly defined. Here we report on the role of the distinctive C-terminal extension present in Apg2 and other metazoan homologs. We found that the first part of this Apg2 subdomain, with propensity to adopt α-helical structure, interacts with the nucleotide binding domain of Hsc70 in a nucleotide-dependent manner, contributing significantly to the stability of the Hsc70:Apg2 complex. Moreover, the second intrinsically disordered segment of Apg2 C-terminal extension plays an important role as a downregulator of nucleotide exchange. An NMR analysis showed that the interaction with Hsc70 nucleotide binding domain modifies the chemical environment of residues located in important functional sites such as the interface between lobe I and II and the nucleotide binding site. Our data indicate that Apg2 C-terminal extension is a fine-tuner of human Hsc70 activity that optimizes the substrate remodeling ability of the chaperone system.
Subject(s)
HSC70 Heat-Shock Proteins , HSP110 Heat-Shock Proteins , Humans , HSC70 Heat-Shock Proteins/chemistry , HSC70 Heat-Shock Proteins/metabolism , HSP110 Heat-Shock Proteins/chemistry , HSP110 Heat-Shock Proteins/metabolism , HSP40 Heat-Shock Proteins/metabolism , HSP70 Heat-Shock Proteins/metabolism , Molecular Chaperones/metabolism , Nucleotides/metabolism , Protein Aggregates , Protein BindingABSTRACT
Costimulation pathways play an essential role in T cell activation, differentiation, and regulation. CD155 expressed on antigen-presenting cells (APCs) interacts with TIGIT, an inhibitory costimulatory molecule, and CD226, an activating costimulatory molecule, on T cells. TIGIT and CD226 are expressed at varying levels depending on the T cell subset and activation state. T follicular helper cells in germinal centers (GC-Tfh) in human tonsils express high TIGIT and low CD226. However, the biological role of the CD155/TIGIT/CD226 axis in human Tfh cell biology has not been elucidated. To address this, we analyzed tonsillar CD4+ T cell subsets cultured with artificial APCs constitutively expressing CD155. Here we show that CD226 signals promote the early phase of Tfh cell differentiation in humans. CD155 signals promoted the proliferation of naïve CD4+ T cells and Tfh precursors (pre-Tfh) isolated from human tonsils and upregulated multiple Tfh molecules and decreased IL-2, a cytokine detrimental for Tfh cell differentiation. Blocking CD226 potently inhibited their proliferation and expression of Tfh markers. By contrast, while CD155 signals promoted the proliferation of tonsillar GC-Tfh cells, their proliferation required only weak CD226 signals. Furthermore, attenuating CD226 signals rather increased the expression of CXCR5, ICOS, and IL-21 by CD155-stimulated GC-Tfh cells. Thus, the importance of CD226 signals changes according to the differentiation stage of human Tfh cells and wanes in mature GC-Tfh cells. High TIGIT expression on GC-Tfh may play a role in attenuating the detrimental CD226 signals post GC-Tfh cell maturation.
Subject(s)
Antigens, Differentiation, T-Lymphocyte , Receptors, Immunologic , T Follicular Helper Cells , Antigens, Differentiation, T-Lymphocyte/metabolism , Cell Differentiation , Humans , Lymphocyte Activation , Receptors, Immunologic/metabolism , T-Lymphocyte SubsetsABSTRACT
Safe and effective vaccines are needed to end the COVID-19 pandemic. Here, we report the preclinical development of a lipid nanoparticleformulated SARS-CoV-2 mRNA vaccine, PTX-COVID19-B. PTX-COVID19-B was chosen among three candidates after the initial mouse vaccination results showed that it elicited the strongest neutralizing antibody response against SARS-CoV-2. Further tests in mice and hamsters indicated that PTX-COVID19-B induced robust humoral and cellular immune responses and completely protected the vaccinated animals from SARS-CoV-2 infection in the lung. Studies in hamsters also showed that PTX-COVID19-B protected the upper respiratory tract from SARS-CoV-2 infection. Mouse immune sera elicited by PTX-COVID19-B vaccination were able to neutralize SARS-CoV-2 variants of concern, including the Alpha, Beta, Gamma, and Delta lineages. No adverse effects were induced by PTX-COVID19-B in either mice or hamsters. Based on these results, PTX-COVID19-B was authorized by Health Canada to enter clinical trials in December 2020 with a phase 2 clinical trial ongoing.
Subject(s)
COVID-19 Vaccines/immunology , COVID-19/prevention & control , SARS-CoV-2/immunology , Spike Glycoprotein, Coronavirus/immunology , Vaccines, Synthetic/immunology , mRNA Vaccines/immunology , Animals , Antibodies, Neutralizing/blood , Antibodies, Viral/blood , CD4 Lymphocyte Count , CD8-Positive T-Lymphocytes/immunology , COVID-19/immunology , COVID-19 Vaccines/adverse effects , Canada , Cell Line , Cricetinae , Drug Evaluation, Preclinical , Female , HEK293 Cells , Humans , Immunity, Cellular/immunology , Immunity, Humoral/immunology , Liposomes/pharmacology , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Nanoparticles , Spike Glycoprotein, Coronavirus/genetics , Th1 Cells/immunologyABSTRACT
The aggregation of α-synuclein is the hallmark of a collective of neurodegenerative disorders known as synucleinopathies. The tendency to aggregate of this protein, the toxicity of its aggregation intermediates and the ability of the cellular protein quality control system to clear these intermediates seems to be regulated, among other factors, by post-translational modifications (PTMs). Among these modifications, we consider herein proteolysis at both the N- and C-terminal regions of α-synuclein as a factor that could modulate disassembly of toxic amyloids by the human disaggregase, a combination of the chaperones Hsc70, DnaJB1 and Apg2. We find that, in contrast to aggregates of the protein lacking the N-terminus, which can be solubilized as efficiently as those of the WT protein, the deletion of the C-terminal domain, either in a recombinant context or as a consequence of calpain treatment, impaired Hsc70-mediated amyloid disassembly. Progressive removal of the negative charges at the C-terminal region induces lateral association of fibrils and type B* oligomers, precluding chaperone action. We propose that truncation-driven aggregate clumping impairs the mechanical action of chaperones, which includes fast protofilament unzipping coupled to depolymerization. Inhibition of the chaperone-mediated clearance of C-truncated species could explain their exacerbated toxicity and higher propensity to deposit found in vivo.
Subject(s)
Amyloid/metabolism , HSP70 Heat-Shock Proteins/metabolism , Protein Aggregation, Pathological/pathology , Synucleinopathies/pathology , alpha-Synuclein/metabolism , Calpain/pharmacology , HSC70 Heat-Shock Proteins/metabolism , HSP40 Heat-Shock Proteins/metabolism , Humans , Molecular Chaperones/metabolism , Protein Aggregates/physiology , Protein Processing, Post-Translational/physiology , ProteolysisABSTRACT
Neurodegenerative diseases (NDs) are increasingly positioned as leading causes of global deaths. The accelerated aging of the population and its strong relationship with neurodegeneration forecast these pathologies as a huge global health problem in the upcoming years. In this scenario, there is an urgent need for understanding the basic molecular mechanisms associated with such diseases. A major molecular hallmark of most NDs is the accumulation of insoluble and toxic protein aggregates, known as amyloids, in extracellular or intracellular deposits. Here, we review the current knowledge on how molecular chaperones, and more specifically a ternary protein complex referred to as the human disaggregase, deals with amyloids. This machinery, composed of the constitutive Hsp70 (Hsc70), the class B J-protein DnaJB1 and the nucleotide exchange factor Apg2 (Hsp110), disassembles amyloids of α-synuclein implicated in Parkinson's disease as well as of other disease-associated proteins such as tau and huntingtin. We highlight recent studies that have led to the dissection of the mechanism used by this chaperone system to perform its disaggregase activity. We also discuss whether this chaperone-mediated disassembly mechanism could be used to solubilize other amyloidogenic substrates. Finally, we evaluate the implications of the chaperone system in amyloid clearance and associated toxicity, which could be critical for the development of new therapies.
Subject(s)
Amyloid/metabolism , Molecular Chaperones/metabolism , Protein Aggregates , Amyloid/toxicity , Humans , Models, Biological , Nerve Degeneration/metabolism , Nerve Degeneration/pathology , alpha-Synuclein/metabolismABSTRACT
α-synuclein aggregation is present in Parkinson's disease and other neuropathologies. Among the assemblies that populate the amyloid formation process, oligomers and short fibrils are the most cytotoxic. The human Hsc70-based disaggregase system can resolve α-synuclein fibrils, but its ability to target other toxic assemblies has not been studied. Here, we show that this chaperone system preferentially disaggregates toxic oligomers and short fibrils, while its activity against large, less toxic amyloids is severely impaired. Biochemical and kinetic characterization of the disassembly process reveals that this behavior is the result of an all-or-none abrupt solubilization of individual aggregates. High-speed atomic force microscopy explicitly shows that disassembly starts with the destabilization of the tips and rapidly progresses to completion through protofilament unzipping and depolymerization without accumulation of harmful oligomeric intermediates. Our data provide molecular insights into the selective processing of toxic amyloids, which is critical to identify potential therapeutic targets against increasingly prevalent neurodegenerative disorders.
Subject(s)
Amyloid/metabolism , Molecular Chaperones/metabolism , alpha-Synuclein/metabolism , Biopolymers/metabolism , Humans , Parkinson Disease/metabolism , Protein AggregatesABSTRACT
There are limited reports of the genetic characterization of Toxoplasma gondii infecting captive macropods in North America. A novel genotype, ToxoDB PCR-RFLP genotype 263, was reported from six wallabies at a zoological facility in Virginia, USA, prompting an investigation into the genotypes from T. gondii strains infecting macropods at a zoological park in Florida, USA. Cardiac muscle and/or lung samples from an agile wallaby (Macropus agilis, n = 1), red kangaroos (Macropus rufus, n = 8), red-necked wallaby (Macropus rufogriseus, n = 1), and a tammar wallaby (Macropus eugenii, n = 1) that died between 2014 and 2018 were collected. All 11 cases were confirmed to have died from systemic toxoplasmosis by histopathology and immunohistochemical staining. Multilocus PCR-RFLP genotyping of T. gondii was performed directly on tissue samples or on parasites isolated from myocardium by mouse bioassay. Two cases of toxoplasmosis were identified as the reported novel genotype, ToxoDB PCR-RFLP genotype 263, but no common source of exposure could be identified. Five cases were identified as genotype 2 (type III strain, haplogroup 3), and four cases were identified as genotype 216, which has been previously reported in North American wildlife. There were no overt differences in lesion severity or distribution related to genotype. These results suggest that the premise was contaminated with at least three genotypes of T. gondii causing systemic toxoplasmosis in macropods. The largest cluster of fatal toxoplasmosis in macropods in the study period occurred following severe rainfall flooding of the exhibit, suggesting the transmission of T. gondii by water and pointing out the importance of this transmission mechanism. In summary, our study revealed three T. gondii outbreaks that caused significant loss of macropods within 5 yr in a zoological facility in Florida. More studies are needed to understand transmission and prevention of toxoplasmosis in sensitive zoo animals.
Subject(s)
Genotype , Macropodidae , Toxoplasma/genetics , Toxoplasmosis, Animal/parasitology , Animals , Animals, Zoo , Florida/epidemiology , Rain , Toxoplasma/isolation & purification , Toxoplasmosis, Animal/mortality , Toxoplasmosis, Animal/transmissionABSTRACT
In the present work, cell lines of different origin were exposed to BPA levels from food intake reported elsewhere. Specifically, we used an in vitro assay to determine cytotoxicity of BPA in three cell lines: MCF7 (breast cancer), PC3 (prostate cancer) and 3T3-L1 (mouse fibroblast). Cytotoxic effects were observed at concentrations higher than 50 µg/mL which is above the involuntary exposure level of BPA described before in fresh, canned and frozen foods and beverages. Furthermore, medial inhibitory concentrations (IC50) of 85.17 µg/mL and 88.48 µg/mL were observed for PC3 and 3T3-L1, respectively, and a slightly lower IC50 of 64.67 µg/mL for MCF7. These results highlight BPA's toxicity potential at current levels from food intake. The cell line-dependent divergent response to BPA reported herein is discussed.
Subject(s)
Benzhydryl Compounds/adverse effects , Benzhydryl Compounds/toxicity , Cell Line/drug effects , Phenols/adverse effects , Phenols/toxicity , 3T3-L1 Cells/drug effects , Animals , Food Contamination , Humans , Inhibitory Concentration 50 , MCF-7 Cells/drug effects , Mice , PC-3 Cells/drug effectsABSTRACT
Objetivo: revisar los niveles del síndrome de burnout en parte del personal asistencialde una clínica oncológica. Materiales y métodos: la investigación fue de tipo descriptiva, la muestra estuvo compuesta por un total de 19 personas, de las cuales 7 son hombres (36.8%) y 12 personas fueron mujeres (63.2%), todos tienen un cargo dentro de la clínica oncológica, se aplicó el M.B.I. y una ficha de caracterización. Resultados: las subescalas del MBI no indican que existe burnout, ya que la mayoría de las personas han puntuando nivel bajo en las escalas de agotamiento emocional y despersonalización, no así para el de realización personal que puntuó alto. Hay un (69%) de personas que no presentan ninguno de los síntomas, en cambio hay un (31,6%) de personas que si manifiestan o puntúan alto o intermedio en algunas de las subescalas agotamiento emocional o despersonalización o baja en realización personal. Conclusiones: no existe evidencia de que las personas que hacen parte del personal asistencial de una clínica oncológica tengan el síndrome de burnout, tampoco hay diferencias significativas entre los grupos asistenciales..(AU)
Objective: to review levels of burnout syndrome in part of the nursing staff of an oncology clinic. Materials and methods: the research was descriptive, the sample was composed of 19 people, seven were men (36.8%) and twelve were women (63.2%), all have a position within the Oncology clinic, It was applied an MBI and a fact sheet of characterization. Results: the subscales of the MBI does not indicate that there is burnout, since most of the people have scoring low on the scales of emotional exhaustion and depersonalization, not for the personal fulfilment that scored high. It shows that (69%) of people do not have any of the symptoms; instead, (31.6%) of people manifested the condition or scored high or intermediate in some of the subscales of emotional exhaustion and depersonalization or low personal fulfilment. Conclusions: there is no evidence that people who are part of the nursing staff of the Oncology clinic have burnout syndrome, nor there are significant differences between the health care groups..(AU)
Subject(s)
Personnel, HospitalABSTRACT
The interaction between tumor and the immune system is still poorly understood. Significant clinical responses have been achieved in cancer patients treated with antibodies against the CTLA4 and PD-1/PD-L1 checkpoints; however, only a small portion of patients responded to the therapies, indicating a need to explore additional co-inhibitory molecules for cancer treatment. B7-H3, a member of the B7 superfamily, was previously shown by us to inhibit T-cell activation and autoimmunity. In this study, we have analyzed the function of B7-H3 in tumor immunity. Expression of B7-H3 was found in multiple tumor lines, tumor-infiltrating dendritic cells, and macrophages. B7-H3-deficient mice or mice treated with an antagonistic antibody to B7-H3 showed reduced growth of multiple tumors, which depended on NK and CD8+ T cells. With a putative receptor expressed by cytotoxic lymphocytes, B7-H3 inhibited their activation, and its deficiency resulted in increased cytotoxic lymphocyte function in tumor-bearing mice. Combining blockades of B7-H3 and PD-1 resulted in further enhanced therapeutic control of late-stage tumors. Taken together, our results indicate that the B7-H3 checkpoint may serve as a novel target for immunotherapy against cancer.
Subject(s)
B7 Antigens/immunology , CD8-Positive T-Lymphocytes/immunology , Immunity, Cellular , Killer Cells, Natural/immunology , Neoplasm Proteins/immunology , Neoplasms/immunology , Animals , B7 Antigens/genetics , CD8-Positive T-Lymphocytes/pathology , Cell Line, Tumor , Female , Humans , Killer Cells, Natural/pathology , Male , Mice , Mice, Inbred BALB C , Mice, Knockout , Neoplasm Proteins/genetics , Neoplasms/genetics , Neoplasms/pathology , Neoplasms/therapyABSTRACT
Environmental Epigenomics is a developing field to study the epigenetic effect on human health from exposure to environmental factors. Endocrine disrupting chemicals have been detected primarily in pharmaceutical drugs, personal care products, food additives, and food containers. Exposure to endocrine-disrupting chemicals (EDCs) has been associated with a high incidence and prevalence of many endocrine-related disorders in humans. Nevertheless, further evidence is needed to establish a correlation between exposure to EDC and human disorders. Conventional detection of EDCs is based on chemical structure and concentration sample analysis. However, substantial evidence has emerged, suggesting that cell exposure to EDCs leads to epigenetic changes, independently of its chemical structure with non-monotonic low-dose responses. Consequently, a paradigm shift in toxicology assessment of EDCs is proposed based on a comprehensive review of analytical techniques used to evaluate the epigenetic effects. Fundamental insights reported elsewhere are compared in order to establish DNA methylation analysis as a viable method for assessing endocrine disruptors beyond the conventional study approach of chemical structure and concentration analysis.
Subject(s)
DNA Methylation/drug effects , Endocrine Disruptors/toxicity , Environmental Exposure , Environmental Pollutants/toxicity , Epigenesis, Genetic/drug effects , Epigenomics , Animals , Endocrine Disruptors/analysis , Environmental Exposure/adverse effects , Environmental Exposure/analysis , Environmental Pollutants/analysis , Epigenesis, Genetic/genetics , Female , Humans , Male , Reproduction/drug effects , Reproduction/geneticsABSTRACT
Wild felids are thought to share parasites with domestic cats. However, little is known of the coccidian parasites of wild felids. We investigated the presence of Sarcocystis spp. in tissues of 6 species of 90 Neotropical small felids killed in road accidents in the state of Rio Grande do Sul, Brazil by using microscopic and molecular techniques. Formalin-fixed tissues from 28 felids were examined, and Sarcocystis felis-like sarcocysts were detected in 4 wild cats (2 Puma yagouaroundi and 2 Leopardus guttulus). By transmission electron microscopy, sarcocysts from a P. yagouaroundi were identical to S. felis from domestic cats in the USA. Direct sequencing of PCR amplicons resulted the unambiguous sequences of the ITS-1 region from 18 of the 31 PCR positive wild cats; 5 sequences from each P. yagouaroundi, and Leopardus geoffroyi, 4 sequences from L. guttulus, and 2 sequences from each Leopardus wiedii, and Leopardus colocolo. Sequences analysis of ITS-1 region revealed the highest identiy (97-99%) with that of previously describe isolates of S. felis from domestic cats in the USA and identified them as S. felis. Tissues of 1 Leopardus pardalis tested by PCR and histology were negative. The phylogenetic relationship indicated that S. felis is quite different to species which employ opossums as their definitive host. This is the first report of S. felis infection in small wild felids from Brazil.
Subject(s)
Felidae/parasitology , Sarcocystis/genetics , Sarcocystis/ultrastructure , Sarcocystosis/parasitology , Animals , Animals, Wild/parasitology , Brazil , Cats , DNA, Ribosomal Spacer/genetics , Microscopy, Electron, Transmission , Phylogeny , Sarcocystis/classification , Sequence Homology, Nucleic Acid , Species Specificity , United StatesABSTRACT
Obesity and metabolic disorders such as insulin resistance and type 2 diabetes have become a major threat to public health globally. The mechanisms that lead to insulin resistance in type 2 diabetes have not been well understood. In this study, we show that mice deficient in MAPK phosphatase 5 (MKP5) develop insulin resistance spontaneously at an early stage of life and glucose intolerance at a later age. Increased macrophage infiltration in white adipose tissue of young MKP5-deficient mice correlates with the development of insulin resistance. Glucose intolerance in MKP5-deficient mice is accompanied by significantly increased visceral adipose weight, reduced AKT activation, enhanced p38 activity, and increased inflammation in visceral adipose tissue when compared with wild-type (WT) mice. Deficiency of MKP5 resulted in increased inflammatory activation in macrophages. These findings thus demonstrate that MKP5 critically controls inflammation in white adipose tissue and the development of metabolic disorders.
Subject(s)
Adipose Tissue/pathology , Inflammation/enzymology , Insulin Resistance , Mitogen-Activated Protein Kinase Phosphatases/metabolism , Adipose Tissue/enzymology , Animals , Glucose/metabolism , Macrophages/pathology , Mice , Mice, Inbred C57BL , Mice, Knockout , Mitogen-Activated Protein Kinase Phosphatases/geneticsABSTRACT
This paper summarizes prevalence of Toxoplasma gondii in humans and animals and associated correlates of infection, clinical spectrum of disease in humans, and genetic diversity of T. gondii isolates from Colombia. Recent studies, especially in the states of Antioquia, Quindío and Cundinamarca, indicate that toxoplasmosis is a major public health problem. Approximately half of the women of child bearing age have T. gondii antibodies, and the clinical disease in congenitally infected children is more severe than in Europe. Limited studies indicate that the strains of T. gondii from Colombia are genetically and phenotypically different than in Europe and North America. However, epidemiological factors, such as the involvement of domestic and/or wild animals in transmission, the distribution of strain diversity by natural geographic regions, and the variation in risk factors between regions that are associated with human infection in Colombia, remain unknown. Areas of research for the future are outlined. This review should be of interest to biologists, veterinarians, physicians, and parasitologists.