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1.
PeerJ ; 12: e17252, 2024.
Article in English | MEDLINE | ID: mdl-38708345

ABSTRACT

Background: Periodontitis is a chronic infectious disease, characterized by an exacerbated inflammatory response and a progressive loss of the supporting tissues of the teeth. Porphyromonas gingivalis is a key etiologic agent in periodontitis. Cystatin C is an antimicrobial salivary peptide that inhibits the growth of P. gingivalis. This study aimed to evaluate the antimicrobial activity of this peptide and its effect on cytokine production, nitric oxide (NO) release, reactive oxygen species (ROS) production, and programmed cell death in human macrophages infected with P. gingivalis. Methods: Monocyte-derived macrophages generated from peripheral blood were infected with P. gingivalis (MOI 1:10) and stimulated with cystatin C (2.75 µg/ml) for 24 h. The intracellular localization of P. gingivalis and cystatin C was determined by immunofluorescence and transmission electron microscopy (TEM). The intracellular antimicrobial activity of cystatin C in macrophages was assessed by counting Colony Forming Units (CFU). ELISA assay was performed to assess inflammatory (TNFα, IL-1ß) and anti-inflammatory (IL-10) cytokines. The production of nitrites and ROS was analyzed by Griess reaction and incubation with 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA), respectively. Programmed cell death was assessed with the TUNEL assay, Annexin-V, and caspase activity was also determined. Results: Our results showed that cystatin C inhibits the extracellular growth of P. gingivalis. In addition, this peptide is internalized in the infected macrophage, decreases the intracellular bacterial load, and reduces the production of inflammatory cytokines and NO. Interestingly, peptide treatment increased ROS production and substantially decreased bacterial-induced macrophage apoptosis. Conclusions: Cystatin C has antimicrobial and immuno-regulatory activity in macrophages infected with P. gingivalis. These findings highlight the importance of understanding the properties of cystatin C for its possible therapeutic use against oral infections such as periodontitis.


Subject(s)
Cystatin C , Macrophages , Nitric Oxide , Porphyromonas gingivalis , Reactive Oxygen Species , Porphyromonas gingivalis/immunology , Humans , Macrophages/immunology , Macrophages/drug effects , Macrophages/metabolism , Macrophages/microbiology , Cystatin C/metabolism , Reactive Oxygen Species/metabolism , Nitric Oxide/metabolism , Cytokines/metabolism , Periodontitis/microbiology , Periodontitis/immunology , Periodontitis/drug therapy , Periodontitis/pathology , Apoptosis/drug effects
2.
FEMS Microbiol Ecol ; 99(11)2023 10 17.
Article in English | MEDLINE | ID: mdl-37827541

ABSTRACT

Important bacterial pathogens such as Pseudomonas aeruginosa produce several exoproducts such as siderophores, degradative enzymes, biosurfactants, and exopolysaccharides that are used extracellularly, benefiting all members of the population, hence being public goods. Since the production of public goods is a cooperative trait, it is in principle susceptible to cheating by individuals in the population who do not invest in their production, but use their benefits, hence increasing their fitness at the expense of the cooperators' fitness. Among the most studied virulence factors susceptible to cheating are siderophores and exoproteases, with several studies in vitro and some in animal infection models. In addition to these two well-known examples, cheating with other virulence factors such as exopolysaccharides, biosurfactants, eDNA production, secretion systems, and biofilm formation has also been studied. In this review, we discuss the evidence of the susceptibility of each of those virulence factors to cheating, as well as the mechanisms that counteract this behavior and the possible consequences for bacterial virulence.


Subject(s)
Siderophores , Virulence Factors , Humans , Virulence Factors/genetics , Pseudomonas aeruginosa/genetics , Biofilms , Quorum Sensing
3.
FEMS Microbiol Ecol ; 99(9)2023 08 22.
Article in English | MEDLINE | ID: mdl-37496200

ABSTRACT

Social cheating is the exploitation of public goods that are costly metabolites, like exoproteases. Exoprotease exploitation in Pseudomonas aeruginosa has been studied in reference strains. Experimental evolution with reference strains during continuous growth in casein has demonstrated that nonexoprotease producers that are lasR mutants are selected while they behave as social cheaters. However, noncanonical quorum-sensing systems exist in P. aeruginosa strains, which are diverse. In this work, the exploitation of exoproteases in the environmental strain ID4365 was evaluated; ID4365 has a nonsense mutation that precludes expression of LasR. ID4365 produces exoproteases under the control of RhlR, and harbors an inducible prophage. As expected, rhlR mutants of ID4365 behave as social cheaters, and exoprotease-deficient individuals accumulate upon continuous growth in casein. Moreover, in all continuous cultures, population collapses occur. However, this also sometimes happens before cheaters dominate. Interestingly, during growth in casein, ID4565's native prophage is induced, suggesting that the metabolic costs imposed by social cheating may increase its induction, promoting population collapses. Accordingly, lysogenization of the PAO1 lasR mutant with this prophage accelerated its collapse. These findings highlight the influence of temperate phages in social cheating.


Subject(s)
Pseudomonas aeruginosa , Quorum Sensing , Humans , Quorum Sensing/genetics , Pseudomonas aeruginosa/genetics , Caseins/genetics , Caseins/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Lysogeny , Prophages/genetics
4.
Cell Transplant ; 32: 9636897231177357, 2023.
Article in English | MEDLINE | ID: mdl-37291807

ABSTRACT

Obesity has been linked to cognitive impairment through systemic low-grade inflammation. High fat and sugar diets (HFSDs) also induce systemic inflammation, either by induced Toll-like receptor 4 response, or by causing dysbiosis. This study aimed to evaluate the effect of symbiotics supplementation on spatial and working memory, butyrate concentration, neurogenesis, and electrophysiological recovery of HFSD-fed rats. In a first experiment, Sprague-Dawley male rats were given HFSD for 10 weeks, after which they were randomized into 2 groups (n = 10 per group): water (control), or Enterococcus faecium + inulin (symbiotic) administration, for 5 weeks. In the fifth week, spatial and working memory was analyzed through the Morris Water Maze (MWM) and Eight-Arm Radial Maze (RAM) tests, respectively, with 1 week apart between tests. At the end of the study, butyrate levels from feces and neurogenesis at hippocampus were determined. In a second experiment with similar characteristics, the hippocampus was extracted to perform electrophysiological studies. Symbiotic-supplemented rats showed a significantly better memory, butyrate concentrations, and neurogenesis. This group also presented an increased firing frequency in hippocampal neurons [and a larger N-methyl-d-aspartate (NMDA)/α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) current ratio] suggesting an increase in NMDA receptors, which in turn is associated with an enhancement in long-term potentiation and synaptic plasticity. Therefore, our results suggest that symbiotics could restore obesity-related memory impairment and promote synaptic plasticity.


Subject(s)
Agave , Spatial Memory , Rats , Animals , Male , Agave/metabolism , Inulin/pharmacology , Inulin/therapeutic use , Rats, Sprague-Dawley , Hippocampus/metabolism , Neuronal Plasticity/physiology , Receptors, N-Methyl-D-Aspartate/metabolism , Maze Learning/physiology , Obesity/therapy , Dietary Supplements , Inflammation
5.
PeerJ ; 11: e15272, 2023.
Article in English | MEDLINE | ID: mdl-37101788

ABSTRACT

Bacteriophages are the most abundant biological entity on the planet, having pivotal roles in bacterial ecology, animal and plant health, and in the biogeochemical cycles. Although, in principle, phages are simple entities that replicate at the expense of their bacterial hosts, due the importance of bacteria in all aspects of nature, they have the potential to influence and modify diverse processes, either in subtle or profound ways. Traditionally, the main application of bacteriophages is phage therapy, which is their utilization to combat and help to clear bacterial infections, from enteric diseases, to skin infections, chronic infections, sepsis, etc. Nevertheless, phages can also be potentially used for several other tasks, including food preservation, disinfection of surfaces, treatment of several dysbioses, and modulation of microbiomes. Phages may also be used as tools for the treatment of non-bacterial infections and pest control in agriculture; moreover, they can be used to decrease bacterial virulence and antibiotic resistance and even to combat global warming. In this review manuscript we discuss these possible applications and promote their implementation.


Subject(s)
Bacterial Infections , Bacteriophages , Phage Therapy , Animals , Bacteria , Bacterial Infections/therapy
6.
Odontology ; 111(4): 778-792, 2023 Oct.
Article in English | MEDLINE | ID: mdl-36897441

ABSTRACT

Porphyromonas gingivalis is a keystone pathogen associated with periodontitis development, a chronic inflammatory pathology characterized by the destruction of the supporting teeth structure. Macrophages are recruited cells in the inflammatory infiltrate from patients with periodontitis. They are activated by the P. gingivalis virulence factors arsenal, promoting an inflammatory microenvironment characterized by cytokine production (TNF-α, IL-1ß, IL-6), prostaglandins, and metalloproteinases (MMPs) that foster the tissular destruction characteristic of periodontitis. Furthermore, P. gingivalis suppresses the generation of nitric oxide, a potent antimicrobial molecule, through its degradation, and incorporating its byproducts as a source of energy. Oral antimicrobial peptides can contribute to controlling the disease due to their antimicrobial and immunoregulatory activity, which allows them to maintain homeostasis in the oral cavity. This study aimed to analyze the immunopathological role of macrophages activated by P. gingivalis in periodontitis and suggested using antimicrobial peptides as therapeutic agents to treat the disease.


Subject(s)
Periodontitis , Porphyromonas gingivalis , Humans , Antimicrobial Peptides , Macrophages/metabolism , Macrophages/pathology , Periodontitis/metabolism , Immunomodulation
7.
Front Cell Infect Microbiol ; 13: 1280265, 2023.
Article in English | MEDLINE | ID: mdl-38298921

ABSTRACT

Background: Bacteriophage therapy is becoming part of mainstream Western medicine since antibiotics of clinical use tend to fail. It involves applying lytic bacteriophages that self-replicate and induce cell lysis, thus killing their hosts. Nevertheless, bacterial killing promotes the selection of resistant clones which sometimes may exhibit a decrease in bacterial virulence or antibiotic resistance. Methods: In this work, we studied the Pseudomonas aeruginosa lytic phage φDCL-PA6 and its variant φDCL-PA6α. Additionally, we characterized and evaluated the production of virulence factors and the virulence in a Galleria mellonella model of resistant mutants against each phage for PA14 and two clinical strains. Results: Phage φDCL-PA6α differs from the original by only two amino acids: one in the baseplate wedge subunit and another in the tail fiber protein. According to genomic data and cross-resistance experiments, these changes may promote the change of the phage receptor from the O-antigen to the core lipopolysaccharide. Interestingly, the host range of the two phages differs as determined against the Pseudomonas aeruginosa reference strains PA14 and PAO1 and against nine multidrug-resistant isolates from ventilator associated pneumonia. Conclusions: We show as well that phage resistance impacts virulence factor production. Specifically, phage resistance led to decreased biofilm formation, swarming, and type III secretion; therefore, the virulence towards Galleria mellonella was dramatically attenuated. Furthermore, antibiotic resistance decreased for one clinical strain. Our study highlights important potential advantages of phage therapy's evolutionary impact that may be exploited to generate robust therapy schemes.


Subject(s)
Bacteriophages , Moths , Phage Therapy , Pseudomonas Phages , Animals , Virulence , Pseudomonas aeruginosa , Pseudomonas Phages/genetics , Virulence Factors/genetics , Drug Resistance, Microbial , Anti-Bacterial Agents/pharmacology
8.
PeerJ ; 10: e14232, 2022.
Article in English | MEDLINE | ID: mdl-36312752

ABSTRACT

Background: Periodontal disease is considered one of the most prevalent chronic infectious diseases, often leading to the disruption of tooth-supporting tissues, including alveolar bone, causing tooth mobility and loss. Porphyromonas gingivalis is considered the major etiological agent of this disease, having a plethora of virulence factors, including, lipopolysaccharides (LPS), hemolysins, and proteinases. Antimicrobial peptides are one of the main components of the innate immune response that inhibit the growth of P. gingivalis. The aim of this study was to analyze the antimicrobial activity of cystatin C and to assess the effect on the inflammatory and anti-inflammatory cytokines, the production of reactive oxygen species, and in the release of nitric oxide by human gingival fibroblasts incubated with P. gingivalis in the presence and absence of cystatin C. Methods: P. gingivalis ATCC 33277 was exposed to cystatin C for 24h and co-cultured with human gingival fibroblasts (HGFs) ATCC CRL-2014. The effect of cystatin on growth of P. gingivalis and HGFs was evaluated. Pro-inflammatory (TNFα, IL-1ß) and anti-inflammatory (IL-10) cytokines were determined by ELISA in the supernatants of HGFs incubated with P. gingivalis exposed to cystatin C. Additionally, nitrites and reactive oxygen species (ROS) production were evaluated. Results: Cystatin Cinhibited the growth of P. gingivalis without affecting HGFs. Incubation of HGFs with P. gingivalis led to a significant increase of TNF-α and IL-1ß. In contrast, HGFs incubated with P. gingivalis exposed to cystatin C showed a decreased production of both cytokines, whereas IL-10 was enhanced. Incubation of HGFs with P. gingivalis led to an increase of nitric oxide (NO) and ROS production, which was reduced in the presence of the peptide. Conclusions: Cystatin C inhibits the growth of P. gingivalis and decreases the inflammatory cytokines, ROS, and NO production during infection of HGFs with P. gingivalis. Knowledge on the antimicrobial and immunomodulatory properties of cystatin C could aid in the design of new therapeutic approaches to facilitate the elimination of this bacterium to improve the treatment of periodontal disease.


Subject(s)
Anti-Infective Agents , Periodontal Diseases , Humans , Porphyromonas gingivalis , Interleukin-10/pharmacology , Reactive Oxygen Species/pharmacology , Cystatin C/pharmacology , Nitric Oxide/pharmacology , Cytokines/pharmacology , Anti-Inflammatory Agents/pharmacology , Anti-Infective Agents/pharmacology , Fibroblasts
9.
PeerJ ; 10: e12667, 2022.
Article in English | MEDLINE | ID: mdl-35116194

ABSTRACT

Antimicrobial peptides (AMPs) are versatile molecules with broad antimicrobial activity produced by representatives of the three domains of life. Also, there are derivatives of AMPs and artificial short peptides that can inhibit microbial growth. Beyond killing microbes, AMPs at grow sub-inhibitory concentrations also exhibit anti-virulence activity against critical pathogenic bacteria, including ESKAPE pathogens. Anti-virulence therapies are an alternative to antibiotics since they do not directly affect viability and growth, and they are considered less likely to generate resistance. Bacterial biofilms significantly increase antibiotic resistance and are linked to establishing chronic infections. Various AMPs can kill biofilm cells and eradicate infections in animal models. However, some can inhibit biofilm formation and promote dispersal at sub-growth inhibitory concentrations. These examples are discussed here, along with those of peptides that inhibit the expression of traits controlled by quorum sensing, such as the production of exoproteases, phenazines, surfactants, toxins, among others. In addition, specific targets that are determinants of virulence include secretion systems (type II, III, and VI) responsible for releasing effector proteins toxic to eukaryotic cells. This review summarizes the current knowledge on the anti-virulence properties of AMPs and the future directions of their research.


Subject(s)
Antimicrobial Cationic Peptides , Antimicrobial Peptides , Animals , Antimicrobial Cationic Peptides/pharmacology , Bacteria/metabolism , Anti-Bacterial Agents/pharmacology
10.
Ther Adv Infect Dis ; 8: 20499361211033715, 2021.
Article in English | MEDLINE | ID: mdl-34408874

ABSTRACT

INTRODUCTION: Chagas disease (CD) is caused by Trypanosoma cruzi. When acquired, the disease develops in stages. For diagnosis, laboratory confirmation is required, and an extensive assessment of the patient's health should be performed. Treatment consists of the administration of trypanocidal drugs, which may cause severe adverse effects. The objective of our systematic review was to analyze data contained in the CD published case reports to understand the challenges that patients and clinicians face worldwide. MATERIALS AND METHODS: We performed a systematic review following the PRISMA guidance. PubMed database was explored using the terms 'American trypanosomiasis' or 'Chagas disease'. Results were limited to human case reports written in English or Spanish. A total of 258 reports (322 patients) were included in the analysis. Metadata was obtained from each article. Following this, it was analyzed to obtain descriptive measures. RESULTS: From the sample, 56.2% were males and 43.8% were females. Most cases were from endemic countries (85.4%). The most common clinical manifestations were fever during the acute stage (70.0%), dyspnea during the chronic stage in its cardiac form (53.7%), and constipation during the chronic stage in its digestive form (73.7%). Most patients were diagnosed in the chronic stage (72.0%). Treatment was administered in 56.2% of cases. The mortality rate for the acute stage cases was 24.4%, while for the chronic stage this was 28.4%. DISCUSSION: CD is a parasitic disease endemic to Latin America, with increasing importance due to human and vector migration. In this review, we report reasons for delays in diagnosis and treatment, and trends in medical practices. Community awareness must be increased to improve CD's diagnoses; health professionals should be appropriately trained to detect and treat infected individuals. Furthermore, public health policies are needed to increase the availability of screening and diagnostic tools, trypanocidal drugs, and, eventually, vaccines.

11.
CNS Neurosci Ther ; 26(6): 650-658, 2020 06.
Article in English | MEDLINE | ID: mdl-32352656

ABSTRACT

AIMS: Immunization with neural-derived peptides (INDP) has demonstrated to be a promising therapy to achieve a regenerative effect in the chronic phase of the spinal cord injury (SCI). Nevertheless, INDP-induced neurogenic effects in the chronic stage of SCI have not been explored. METHODS AND RESULTS: In this study, we analyzed the effect of INDP on both motor and sensitive function recovery; afterward, we assessed neurogenesis and determined the production of cytokines (IL-4, IL-10, and TNF alpha) and neurotrophic factors (BDNF and GAP-43). During the chronic stage of SCI, rats subjected to INDP showed a significant increase in both motor and sensitive recovery when compared to the control group. Moreover, we found a significant increase in neurogenesis, mainly at the central canal and at both the dorsal and ventral horns of INDP-treated animals. Finally, INDP induced significant production of antiinflammatory and regeneration-associated proteins in the chronic stages of SCI. CONCLUSIONS: These findings suggest that INDP has a neurogenic effect that could improve motor and sensitive recovery in the chronic stage of SCI. Moreover, our results also envision the use of INDP as a possible therapeutic strategy for other trauma-related disorders like traumatic brain injury.


Subject(s)
Immunization/methods , Neurogenesis/drug effects , Neuropeptides/administration & dosage , Spinal Cord Injuries/metabolism , Spinal Cord Injuries/therapy , Animals , Female , Inflammation Mediators/antagonists & inhibitors , Inflammation Mediators/metabolism , Neurogenesis/physiology , Pain Measurement/drug effects , Pain Measurement/methods , Rats , Rats, Sprague-Dawley , Spinal Cord Injuries/immunology
13.
Curr Opin Pharmacol ; 48: 48-56, 2019 10.
Article in English | MEDLINE | ID: mdl-31136908

ABSTRACT

As current levels of antimicrobial resistance are alarming, the World Health Organization urged the development of new antimicrobials to fight infections produced by multidrug resistant bacteria. Antibiotics impose severe selective pressure for the development of resistance, and currently bacteria resistant to all of them exist. In this review, we discuss the release and development of new antibacterial drugs and their properties as well as the current advances in the development of alternative approaches to combat bacterial infections, including the repurposing of drugs, anti-virulence therapies, the use of photosensitizers, phage therapy, and immunotherapies, with an emphasis on what is currently known about the possible development of bacterial resistance against them.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Bacterial Infections/drug therapy , Drug Resistance, Bacterial , Animals , Drug Utilization , Humans
14.
Front Aging Neurosci ; 10: 416, 2018.
Article in English | MEDLINE | ID: mdl-30618722

ABSTRACT

Aging is associated with morphological, physiological and metabolic changes, leading to multiorgan degenerative pathologies, such as cognitive function decline. It has been suggested that memory loss also involves a decrease in neurotrophic factors, including brain-derived neurotrophic factor (BDNF). In recent years, microbiota has been proposed as an essential player in brain development, as it is believed to activate BDNF secretion through butyrate production. Thus, microbiota modulation by supplementation with probiotics and prebiotics may impact cognitive decline. This study aimed to evaluate the effects of probiotics and prebiotics supplementation on the memory of middle-aged rats. Sprague-Dawley male rats were randomized in four groups (n = 13 per group): control (water), probiotic (E. faecium), prebiotic (agave inulin), symbiotic (E. faecium + inulin), which were administered for 5 weeks by oral gavage. Spatial and associative memory was analyzed using the Morris Water Maze (MWM) and Pavlovian autoshaping tests, respectively. Hippocampus was obtained to analyze cytokines [interleukin (IL-1ß) and tumor necrosis factor (TNF-α)], BDNF and γ-aminobutyric acid (GABA) by enzyme-linked immunosorbent assay (ELISA). Butyrate concentrations were also evaluated in feces. The symbiotic group showed a significantly better performance in MWM (p < 0.01), but not in Pavlovian autoshaping test. It also showed significantly lower concentrations of pro-inflammatory cytokines (p < 0.01) and the reduction in IL-1ß correlated with a better performance of the symbiotic group in MWM (p < 0.05). Symbiotic group also showed the highest BDNF and butyrate levels (p < 0.0001). Finally, we compared the electrophysiological responses of control (n = 8) and symbiotic (n = 8) groups. Passive properties of CA1 pyramidal cells (PCs) exhibited changes in response to the symbiotic treatment. Likewise, this group showed an increase in the N-methyl-D-aspartate receptor (NMDA)/AMPA ratio and exhibited robust long-term potentiation (LTP; p < 0.01). Integrated results suggest that symbiotics could improve age-related impaired memory.

15.
Rev Inst Med Trop Sao Paulo ; 59: e46, 2017 Aug 07.
Article in English | MEDLINE | ID: mdl-28793017

ABSTRACT

Electron microscopy is routinely used to identify viral infections in protozoan parasites. These viruses have been described as non-enveloped and icosahedral structures with a diameter of 30-60 nm. Most of them are classified within the non-segmented dsRNA Totiviridae family. We observed virus-like particles (VLPs) through transmission electron microscopy in the cytoplasm of Trypanosoma cruzi epimastigotes grown in cultures. Clusters of electrodense enveloped VLPs having a diameter of 48 nm were also observed. These clusters appear to have been released from distended Golgi cisternae. Furthermore, a paracrystalline array of electrodense, non-enveloped VLPs (with a diameter of 32 nm) were found in distended Golgi cisternae or as smaller clusters at a distance from the RE or Golgi. We cannot rule out that the 48 nm enveloped VLPs belong to the ssRNA Flaviviridae family because they are within its size range. The localization of enveloped VLPs is consistent with the replication strategy of these viruses that transit through the Golgi to be released at the cell surface. Due to the size and shape of the 32 nm non-enveloped VLPs, we propose that they belong to the dsRNA Totiviridae family. This is the first description of cytoplasmic enveloped and non-enveloped VLPs in T. cruzi epimastigotes.


Subject(s)
Trypanosoma cruzi/virology , Virion , Animals , Mice , Microscopy, Electron, Transmission , Trypanosoma cruzi/ultrastructure
16.
BMC Neurosci ; 18(1): 7, 2017 01 05.
Article in English | MEDLINE | ID: mdl-28056790

ABSTRACT

BACKGROUND: Immunization with neural derived peptides (INDP) as well as scar removal-separately-have shown to induce morphological and functional improvement after spinal cord injury (SCI). In the present study, we compared the effect of INDP alone versus INDP with scar removal on motor recovery, regeneration-associated and cytokine gene expression, and axonal regeneration after chronic SCI. Scar removal was conducted through a single incision with a double-bladed scalpel along the stump, and scar renewal was halted by adding α,α'-dipyridyl. RESULTS: During the chronic injury stage, two experiments were undertaken. The first experiment was aimed at testing the therapeutic effect of INDP combined with scar removal. Sixty days after therapeutic intervention, the expression of genes encoding for TNFα, IFNγ, IL4, TGFß, BDNF, IGF1, and GAP43 was evaluated at the site of injury. Tyrosine hydroxylase and 5-hydroxytryptamine positive fibers were also studied. Locomotor evaluations showed a significant recovery in the group treated with scar removal + INDP. Moreover; this group presented a significant increase in IL4, TGFß, BDNF, IGF1, and GAP43 expression, but a decrease of TNFα and IFNγ. Also, the spinal cord of animals receiving both treatments presented a significant increase of serotonergic and catecholaminergic fibers as compared to other the groups. The second experiment compared the results of the combined approach versus INDP alone. Rats receiving INDP likewise showed improved motor recovery, although on a lesser scale than those who received the combined treatment. An increase in inflammation and regeneration-associated gene expression, as well as in the percentage of serotonergic and catecholaminergic fibers was observed in INDP-treated rats to a lesser degree than those in the combined therapy group. CONCLUSIONS: These findings suggest that INDP, both alone and in combination with scar removal, could modify the non-permissive microenvironment prevailing at the chronic phase of SCI, providing the opportunity of improving motor recovery.


Subject(s)
Cicatrix/metabolism , Locomotion/drug effects , Neuropeptides/administration & dosage , Spinal Cord Injuries/immunology , Spinal Cord Injuries/metabolism , Vaccination , Animals , Brain-Derived Neurotrophic Factor/metabolism , Chronic Disease , Cytokines/metabolism , Disease Models, Animal , Female , GAP-43 Protein/metabolism , Insulin-Like Growth Factor I/metabolism , Interferon-gamma/metabolism , Interleukin-4/metabolism , Neuropeptides/therapeutic use , Rats , Rats, Sprague-Dawley , Recovery of Function , Spinal Cord Injuries/drug therapy , Spinal Cord Regeneration/genetics , Transforming Growth Factor beta/metabolism , Tumor Necrosis Factor-alpha/metabolism
17.
Parasitol Res ; 106(6): 1327-37, 2010 May.
Article in English | MEDLINE | ID: mdl-20237802

ABSTRACT

The susceptibility of Trypanosoma cruzi epimastigotes to lysis by normal or immune sera in a complement-dependent reaction has been reported. Mouse immune sera depleted complement-induced damage in epimastigotes characterized by morphological changes and death. The purpose of this work was to study the mechanism of death in epimastigotes exposed to decomplemented mouse immune serum. Epimastigotes were maintained in RPMI medium. Immune sera were prepared in mice by immunization with whole crude epimastigote extracts. Viable epimastigotes were incubated with decomplemented normal or immune sera at 37 degrees C. By electron microscopy, agglutinated parasites showed characteristic patterns of membrane fusion between two or more parasites; this fusion also produced interdigitation of the subpellicular microtubules. Apoptosis was determined by flow cytometry using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and annexin V assays. Nuclear features were examined by 4'-,6-diamidino-2'-phenylindole diHCI cytochemistry that demonstrated apoptotic nuclear condensation. Caspase activity was also measured. TUNEL results showed that parasites incubated with decomplemented immune sera took up 26% of specific fluorescence as compared to 1.3% in parasites incubated with decomplemented normal sera. The Annexin-V-Fluos staining kit revealed that epimastigotes incubated with decomplemented immune sera exposed phosphatidylserine on the external leaflet of the plasma membrane. The incubation of parasites with immune sera showed caspase 3 activity. We conclude that specific antibodies are able to induce agglutination and apoptosis in epimastigotes, although the pathway is not elucidated.


Subject(s)
Antibodies, Protozoan/immunology , Apoptosis , Complement System Proteins/immunology , Trypanosoma cruzi/immunology , Animals , Annexin A5/analysis , Caspase 3/analysis , Female , In Situ Nick-End Labeling , Mice , Microbial Viability , Microscopy, Electron , Trypanosoma cruzi/chemistry , Trypanosoma cruzi/ultrastructure
18.
Parasitol Res ; 103(4): 847-52, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18626663

ABSTRACT

Larval and adult stages of Taenia solium and Taenia crassiceps WFU strain were analyzed by histochemical and biochemical methods to determine the existence of steroid pathways. The presence of the key enzyme 3beta-hydroxisteroid-dehydrogenase (3beta-HSD) was examined in frozen sections of cysticerci obtained from mice and segments of tapeworms obtained from the intestine of hamsters. 3beta-HSD activity was detected by nitroblue-tetrazolium products after incubation with dehydroepiandrosterone, androstendiol, or pregnenolone. Tapeworm tissues exhibited 3beta-HSD activity in the subtegumentary areas of the neck and immature proglottids following incubation with androstendiol, as well as surrounding the testes in mature proglottids. T. solium cysticerci exhibited 3beta-HSD activity in the subtegumentary tissues. The synthesis of steroid hormones involving the activity of 3beta-HSD was studied in cysticerci or tapeworms incubated in the presence of tritiated steroid precursors. The culture media were analyzed by thin layer chromatography and showed synthesis of androstendiol, testosterone, and 17beta-estradiol by cysticerci, androstendiol, and 17beta-estradiol by tapeworms. The results strongly suggest the activity of 3beta-HSD in taeniid parasites that have at least a part of the enzymatic chain required for androgen and estrogen synthesis and that the enzymes are present in the larval stage and from the early strobilar stages to the mature proglottids.


Subject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Cysticercus/enzymology , Taenia/enzymology , Androstenediol/metabolism , Animals , Chromatography, Thin Layer , Cricetinae , Culture Media/chemistry , Cysticercus/growth & development , Dehydroepiandrosterone/metabolism , Estradiol/analysis , Intestines/parasitology , Mice , Mice, Inbred BALB C , Nitroblue Tetrazolium/metabolism , Pregnenolone/metabolism , Staining and Labeling , Taenia/growth & development , Testosterone/analysis
19.
Vet. Méx ; 32(2): 141-144, abr.-jun. 2001. tab, CD-ROM
Article in Spanish | LILACS | ID: lil-303177

ABSTRACT

Se realizó un estudio transversal de frecuencia con el objetivo de buscar larvas enquistadas viables de Trichinella spiralis en carne de cerdo que se comercializa en mercados públicos, carnicerías y tianguis (mercados ambulantes) de la ciudad de México. Se examinaron 500 muestras de carne de cerdo de l00 g cada una, 165 se obtuvieron en carnicerías establecidas, 208 en mercados públicos y 127 en tianguis ubicados en las 16 delegaciones políticas que conforman la ciudad de México. De cada muestra de carne se tomaron cinco fragmentos a diferentes niveles, todas las muestras fueron de 1 g cada una para su examen mediante triquinoscopía utilizando microscopios Carl Zeiss con objetivos seco-débil y seco-fuerte. Los 2 500 cortes de carne de cerdo examinados resultaron negativos a la presencia de larvas de Trichinella spiralis. Este resultado sugiere que es mínimo el riesgo de adquisición de triquinelosis para el consumidor de carne de cerdo que se expende en la ciudad de México.


Subject(s)
Swine , Trichinella spiralis , Meat , In Vitro Techniques , Zoonoses
20.
Rev. mex. patol. clín ; 47(3): 156-161, jul.-sept. 2000. tab, CD-ROM
Article in Spanish | LILACS | ID: lil-304252

ABSTRACT

La triquinelosis constituye una enfermedad zoonótica cosmopolita de origen alimentario, la causa el nemátodo Trichinella spiralis. En México se ignora su frecuencia a pesar de los numerosos brotes epidémicos que se han presentado. El presente trabajo tiene como objetivo determinar, mediante hemaglutinación indirecta (HAI), la prevalencia de anticuerpos específicos contra antígenos de Trichinella spiralis, que se encontraron en el suero de una muestra de escolares que viven en la ciudad de México. Se analizaron 211 sueros de escolares de seis a doce años de edad, 111 (52.6 por ciento) varones y 110 (47.4 por ciento) niñas. Se formaron seis grupos de estudio, uno por cada grado escolar. El análisis inmunológico de los sueros se realizó con la microtécnica de hemaglutinación indirecta (HAI).Siete (3.3 por ciento) de los 211 sueros analizados resultaron positivos con títulos de dilución de 1:32; cinco sueros correspondieron a alumnos de 10 años de edad que cursaban el quinto grado escolar: cuatro varones y una niña; los otros dos sueros pertenecieron a niñas de nueve años de edad, del cuarto grado escolar. No se observó diferencia de reactividad respecto del sexo.La seropositividad obtenida en este estudio (3.3 por ciento) constituye un indicador de que la transmisión de triquinelosis está afectando a los habitantes de la población estudiada. Se analizan las posibles causas que favorecen esa transmisión.


Subject(s)
Humans , Male , Female , Trichinellosis , Trichinella spiralis , Mexico , Food Contamination , Child , Zoonoses , Hemagglutination Tests/methods , Immunologic Tests/methods
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