Premature Aging in Chronic Kidney Disease: The Outcome of Persistent Inflammation beyond the Bounds.

Over the last hundred years, life expectancy in developed countries has increased because of healthier living habits and the treatment of chronic pathologies causing premature aging. Aging is an inexorable, time-dependent, multifactorial process characterized by a series of progressive and irreversible physiological changes associated with loss of functional, psychological, and social capabilities. Numerous factors, such as oxidative stress, inflammation, and cellular senescence, and an irreversible geriatric syndrome known as frailty, contribute to human body deterioration in aging. The speed of aging may differ between individuals depending on the presence or absence of multiple factors (genetic and/or environment) and the subsequent misbalance of homeostasis, together with the increase of frailty, which also plays a key role in developing chronic diseases. In addition, pathological circumstances have been reported to precipitate or accelerate the aging process. This review investigated the mechanisms involved in the developing pathologies, particularly chronic kidney disease, associated with aging.

Enveloped and non-enveloped viral-like particles in Trypanosoma cruzi epimastigotes.

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.

Cryptic patterning of avian skin confers a developmental facility for loss of neck feathering.

Vertebrate skin is characterized by its patterned array of appendages, whether feathers, hairs, or scales. In avian skin the distribution of feathers occurs on two distinct spatial levels. Grouping of feathers within discrete tracts, with bare skin lying between the tracts, is termed the macropattern, while the smaller scale periodic spacing between individual feathers is referred to as the micropattern. The degree of integration between the patterning mechanisms that operate on these two scales during development and the mechanisms underlying the remarkable evolvability of skin macropatterns are unknown. A striking example of macropattern variation is the convergent loss of neck feathering in multiple species, a trait associated with heat tolerance in both wild and domestic birds. In chicken, a mutation called Naked neck is characterized by a reduction of body feathering and completely bare neck. Here we perform genetic fine mapping of the causative region and identify a large insertion associated with the Naked neck trait. A strong candidate gene in the critical interval, BMP12/GDF7, displays markedly elevated expression in Naked neck embryonic skin due to a cis-regulatory effect of the causative mutation. BMP family members inhibit embryonic feather formation by acting in a reaction-diffusion mechanism, and we find that selective production of retinoic acid by neck skin potentiates BMP signaling, making neck skin more sensitive than body skin to suppression of feather development. This selective production of retinoic acid by neck skin constitutes a cryptic pattern as its effects on feathering are not revealed until gross BMP levels are altered. This developmental modularity of neck and body skin allows simple quantitative changes in BMP levels to produce a sparsely feathered or bare neck while maintaining robust feather patterning on the body.

Taenia crassiceps: A secretion-substance of low molecular weight leads to disruption and apoptosis of seminiferous epithelium cells in male mice.

The present research was performed to isolate and study the effects of a low molecular weight (<1300Da) parasite-associated substance, obtained from peritoneal fluids of female mice infected with Taenia crassiceps cysticerci, on seminiferous epithelium cells of male mice testis. The results showed an intense disruption of Sertoli cells and germ cells within the seminiferous tubules of experimental mice, along with the destruction of their gap junction (GJ). Significant generalized apoptosis of germ cells within seminiferous tubules was determined by TUNEL staining (P=0.0159). In addition, a significant number of infiltrating macrophages were found in the luminal space of these seminiferous tubules (P<0.0001). Finally, electron microscopy studies revealed structural and morphological abnormalities in the somatic cells (Sertoli and Leydig cells) and in the germ cells, primarily in the round and elongate spermatids.

Taenia crassiceps: infections of male mice lead to severe disruption of seminiferous tubule cells and increased apoptosis.

This research was carried out to study the effects of infection with Taenia crassiceps cysticerci on the seminiferous epithelium histoarchitecture in the testes of male mice. Our results showed a severe disruption of the histoarchitecture of the testis epithelium in infected mice. In these animals, a significant infiltration of macrophages within seminiferous tubules was observed (P < 0.001). Generalized apoptosis of germ cells within the seminiferous tubules was observed, as assessed by TUNEL assay and apoptotic nuclei were quantified. The total number of fluorescent objects (DNA) (including clusters, singles, and objects in clusters) was significantly higher in the infected cells than in the control group (P = 0.0286). Observation of the interstitial tissue showed disorder and deterioration of many Leydig cells of infected mice, as well as intense vacuolization and destruction of their inter-cellular junctions. Several ultrastructural abnormalities were observed through electron microscopy as well. The observed pathology could lead to a state of infertility.

Decrease of peritoneal inflammatory CD4(+), CD8(+), CD19(+) lymphocytes and apoptosis of eosinophils in a murine Taenia crassiceps infection.

After an intraperitoneal infection of mice with Taenia crassiceps metacestodes, peritoneal inflammatory cells labeled with fluoresceinated MoAb anti-mouse were analyzed by flow cytometry. Apoptosis was studied by annexin A/PI, TUNEL assays, DNA laddering, caspase-3 activity, and electron microscopy. An important continuous decrease of CD4+, CD8+ and CD19+ lymphocytes, and an increase of eosinophils and macrophages throughout the observation time were found. Apoptosis of eosinophils was quantified during the observation period with a peak at 6 days post-infection (67.27%). In an additional experiment at 12 days post-infection using TUNEL staining, a high level of apoptosis of eosinophil (92.3%) and a significant decrease of CD4+, CD8+, and CD19+ lymphocytes were confirmed. Caspase-3 activity in peritoneal fluid, peritoneal cells' DNA fragmentation, and apoptosis of eosinophils and monocytes were found. The dramatic decrease of peritoneal inflammatory T and B cells and the high level of apoptosis of inflammatory eosinophils induced in mice by infection with T. crassiceps cysticerci may be important factors of the immunosuppression observed in cysticercosis.

Specific antibodies induce apoptosis in Trypanosoma cruzi epimastigotes.

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.

Lymphocyte apoptosis in the inflammatory reaction around Taenia solium metacestodes in porcine cysticercosis.

In the current research, we report apoptosis of lymphocytes in the inflammatory reaction around metacestodes in muscle tissue from cysticercotic pigs. Two events, high metacestode viability (100%) and high cysteine protease activity were found to be closely related to a high phosphatydilserine expression by inflammatory lymphocytes (56%). Testing the RPMI medium used for washing away inflammatory cells from metacestodes with 100% viability, with the fluorescent substrate Z-Phe-Ala-AFC for measuring cysteine protease activity, significant fluorescent values were found. In contrast, tests performed with RPMI medium used for washing away inflammatory cells from metacestodes with 90% viability or less, showed low fluorescence values. Flow cytometry analyses of inflammatory cells obtained from four naturally cysticercotic pigs, and stained with Annexin-V/PI, showed lymphocytes expressing phosphatidylserine with values of 0, 6, 41 and 56% on their outer surfaces. Electron microscopy studies of inflammatory cells from metacestodes with 100% viability, showed lymphocytes with strangled and fragmented nuclei, and heterochromatin displaced to the nuclear periphery. In addition, DNA from these cells showed fragmentation in electrophoresis assays. Apoptosis of lymphocytes in the inflammatory reaction around Taenia solium metacestodes, might have been induced by the parasite cysteine protease, and may be involved in impairing cell-mediated immune responses in human and porcine cysticercosis.

The implantation of Taenia solium metacestodes in mice induces down-modulation of T-cell proliferation and cytokine production.

The purpose of this study was to determine the effect of the implantation of Taenia solium metacestodes and the treatment with suppressive metacestode factor (F1) on the ability of spleen cells from Balb/c mice to produce cytokines. Cytokine production was estimated 12 days following the implantation or 4 days after the last dose of F1 (five doses) by RT-PCR and flow cytometry analyses. Spleen cells were obtained from metacestode-implanted, F1-treated and control mice. They were stimulated with concanavalin A (ConA) ex vivo and used for RT-PCR studies and for CD25 expression and intracellular cytokine production estimations using specific monoclonal antibodies labeled with phycoerithrin or fluorescein. Results of the RT-PCR showed that all cells expressed IFN-gamma, IL-2 and IL-4 mRNAs. IL-10 mRNA was not expressed in any case. Flow cytometry analyses showed that both spleen CD4+ and CD8+ cells from metacestode-implanted or treated-F1 mice expressed significantly diminished percentages of CD25 when compared with control cells (P<0.05). The estimation of intracellular cytokines showed that the production of IL-2 and IL-4 in CD8+ cells, and of IFN-gamma in CD4+ cells from mice implanted with metacestodes was significantly impaired when compared with the values from control cells (P<0.05).

Taenia solium metacestode antigens which are protective for pigs induce Th1/Th2 mixed responses in mice.

The purpose of this study was to determine the Th1 and Th2 cytokine responses induced by Taenia solium metacestode antigens in mice and correlate them with the immune responses elicited in vivo. To assess this aim, mice were inoculated with metacestode antigens. RNA was obtained from spleen cells of immunized or control mice incubated with metacestode antigens and used to determine the cytokine profile. Peripheral blood eosinophilia was measured daily in each mouse and specific serum antibody levels were determined. Results showed that metacestode antigens induce the synthesis of IL-4, IL-5 and IFN-gamma mRNAs in spleen cells. They also induced peripheral blood eosinophilia and elicited specific IgE and IgG antibodies, especially IgG1. Three antigens were recognized by all IgG subclasses and by IgE (104, 88 and 7 kDa), and a 57-kDa protein was recognized by IgG1, IgG2a, IgG2b, and IgE. IgG1 and IgG2b recognized 52, 30 and 20 kDa antigens. Immune responses elicited in vivo and the cytokine profile showed good correlation.

Comparison of the immune responses against Salmonella enterica serovar Gallinarum infection between naked neck chickens and a commercial chicken line.

The immune responses of indigenous naked neck (NaNa and Nana) and normally feathered (nana) chickens against a Salmonella Gallinarum (SG) infection were evaluated and compared with those of a commercial line (B-380). Groups of 28-day-old chickens (NaNa, Nana, nana, and B-380) were immunized orally and subcutaneously with 50 microg of SG antigens. Control non-immunized animals were inoculated with sterile saline solution. All chickens were challenged with 1 LD(50) of SG and mortality was recorded daily for 20 days. Antibodies to SG were measured in sera before immunization, before the challenge, 10 days after the challenge, and at sacrifice. Peripheral blood lymphocyte proliferation assays were performed using concanavalin A and SG antigens. Results showed that non-immunized Nana chickens exhibited the best natural resistance to Salmonella infection, since only 30% of them died. In contrast, all control B-380 chickens died by the 13th day. Immunization with SG induced immunity in chickens of all genotypes. Indigenous naked neck and normally feathered chickens showed a higher survival rate when compared with B-380 chickens. Immunized Nana chickens showed the highest antibody titres (P<0.05) as well as the highest thymidine incorporation in peripheral blood lymphocytes stimulated with con A or SG antigens (P<0.05). The results show that Nana chickens are the most resistant to SG infection and the best responders to vaccination with SG antigens.

Induction of DNA damage in human lymphocytes treated with a soluble factor secreted by Taenia solium metacestodes.

We have previously reported that a factor secreted by the metacestode of Taenia solium (MF) is able to transform Syrian hamster embryo cells. The aim of this study was to analyze the genotoxicity of MF in cultured human lymphocytes using the micronucleus assay. Results show a significantly high frequency of micronucleated cells in lymphocyte cultures treated with MF. Although further experiments are needed to determine whether this factor is also secreted by T. solium metacestodes in humans, analysis of the frequency of micronucleus induced in cultured human lymphocytes indicates that DNA instability induced by MF could represent a risk for malignant transformation.

Discrimination between active and inactive neurocysticercosis by metacestode excretory/secretory antigens of Taenia solium in an enzyme-linked immunosorbent assay.

To detect IgG antibodies to Taenia solium, a controlled double-blind study was conducted using 91 coded cerebrospinal fluid samples from patients with neurocysticercosis (NCC) and other neurologic disorders. Samples were tested in an enzyme-linked immunosorbent assay (ELISA) using metacestode excretion/secretion antigens. The results were correlated with data from medical records on the diagnosis of NCC (based on computed tomography and magnetic resonance imaging criteria) and other neurologic disorders. The ELISA results were positive in 22 of the 24 cases with active NCC. In contrast, six cases with calcified cysts (inactive NCC), as well as one case in a transitional stage, were negative. One case with a calcified granuloma and another with a granuloma plus calcifications (classified as inactive NCC) had positive results. The remaining negative results corresponded to other neurologic disorders (58 cases). The results of the ELISA showed a significant difference between active and inactive NCC (P = 0.0034).

Immunodepresión de linfocitos T en cerdos, modulada pro Cysticercus cellulosae / Immunodepression of T-lymphocytes in hogs modulated by Cysticercus cellulosae

Por el uso de anticuerpos monoclonales fluoresceinados se descubrió que en el 50% de cerdos testigos no inmunizados que habian sido infectados con 20,000 huevos de Taenia solium, el número de linfocitos T y subpoblación T cooperadora (OKT 4) fue más bajo que el normal; mientras que en 83% de cerdos inmunizados y en resto de los testigos se observaron valores normales. En los cerdos disminuidos de linfocitos T, el número de cisticercos implantados fue no sólo 2.18 veces más alto que en los cerdos inmunes sino también 2.4 veces más alto que en el resto de los cerdos testigos. Esto sugiere que se requiere un determinado número de larvas implantadas para inducir inmunodepresión de linfocitos T, en particular células T cooperadoras. Más aún en el cerdo testigo más inmunodeprimido, los linfocitos B también estaban disminuidos. En contraste, en solo un cerdo de los inmunizados (16%) los linfocitos T totales y T cooperadores estaban ligeramente disminuidos, un gran número de cisticercos (244) fue encontrado en este animal, lo que apoya la idea de que una carga parasitaria importante podría infuenciar la regulación inmune. Una intensa reacción granulomatosa rodeando a los cisticercos se observó en cerdos inmunizados. Los eosinófilos fueron las células predominantes y las que se observaran en estrecho contacto e infiltrando las estructuras larvales, también se observaron gránulos de eosinófilos en los canales espirales y adheridos íntimamente a las superficies externas del tegumento larval. Los tegumentos de la vesícula y del gusano mostraron signos evidentes de destrucción

Efecto de la inmunización en cerdos inmunodeprimidos, naturalmente parasitados con Cysticercus cellulosae / Effect of the immunization on immunodepressed hogs, infected naturally by Cysticercus cellulosae

Se encontró inmunodepresión de linfocitos T y B en cerdos naturalmente parasitados por cisticercos de Taenia solium. La inmunodepresión fue proporcional a la mgnitud de la carga parasitaria. El regreso hacia niveles normales en 2 de 3 cerdos fue inducido por inmunización con antígenos de cisticerco, la inmunidad se estimó en función de la incapacidad de las larvas para evaginar. En tanto que el 93% de cisticercos obtenidos de carne parasitada confiscada en rastros de la ciudad de México evaginaron completamente mostrando una movilidad muy activa y prolongada, en un cerdo con 700 cisticercos por kilogramo de tejido muscular ninguna larva se encontró viable, mientras que en un segundo, que tenía 2.100 cisticercos por kilo de tejido parasitado, el 34% de las larvas evaginaron con mucha dificultad y solo parcialmente. En contraste, en el tercer cerdo la inmunodepresión fue progresiva e irreversible; este animal tenía 300 larvas/Kg, de las cuales el 93% evaginó completamente, lo que sugirió que no se indujo inmunidad en este caso