118 research outputs found
Only complete rejoining of DNA strand breaks after UVC allows K562 cell proliferation and DMSO induction of erythropoiesis
DNA strand breaks are early intermediates of the repair of UVC-induced DNA damage, however, since they severely impair cellular activities, their presence should be limited in time. In this study, the effects of incomplete repair of UVC-induced DNA strand breaks are investigated on K562 cell growth and the induction of erythroid differentiation by addition of DMSO to the cell culture medium. The kinetics were followed after UV irradiation by single cell gel electrophoresis, and in total cell population by alkaline or neutral agarose gel electrophoresis. Shortly after exposure, an extensive fragmentation occurred in DNA; DNA double strand breaks were negatively correlated with recovery time for DNA integrity. DNA damage induced by UVC 9 J/m2 rapidly triggered necrosis in a large fraction of irradiated K562 cells, and only 40% of treated cells resumed growth at a very low rate within 24 h of culture. The addition of DMSO to the culture medium of cells 15 min after UVC, when DNA strand break repair was not yet complete, produced apoptosis in >70% of surviving cells, as determined by TUNEL assay. Conversely, if DMSO was added when the resealing of DNA strand breaks was complete, surviving K562 cells retained full growth capacity, and their progeny underwent erythroid differentiation with normal levels of erythroid proteins, d-aminolevulinic acid dehydrase and hemoglobin. This study shows that the extent of DNA strand break repair influences cell proliferation and the DMSO induced erythroid program,and the same UVC dose can have opposite effects depending on cellular status
Apoptosis during chick inner ear development: some observations by TEM and TUNEL techniques.
In order to clarify the occurrence, distribution and possible role of apoptosis during inner ear development, the ultrastructural aspects (by TEM) (at 9-19 incubation day and 1 day after hatching) and the distribution of the apoptotic phenomenon (by the TdTmediated dUTP nick end-labeling technique), were studied in the crista ampullaris of chick embryo at 5-19 days of incubation to hatching and of postnatal 1-day old chick. We found, in the sensorial epithelium, dark supporting cells in chick embryos and mainly dark hair cells in postnatal chicks, both with ultrastructural features consistent with those of apoptosis. The presence of apoptotic phenomena was confirmed by the TUNEL technique. According to our findings, it is hypothesized that apoptosis in the inner ear may be involved: 1) at first, in macroscopic remodelling of the membranous labyrinth in early developmental stages, 2) later, in the correct differentiation of the hair and of the supporting cells, leading to characteristic cellular pattern formation and 3) finally, in physiological cell turnover of the postnatal chicken sensorial epithelium of the crista
Ofd1 Controls Dorso-Ventral Patterning and Axoneme Elongation during Embryonic Brain Development.
Oral-facial-digital type I syndrome (OFDI) is a human X-linked dominant-male-lethal developmental disorder caused by mutations in the OFD1 gene. Similar to other inherited disorders associated to ciliary dysfunction OFD type I patients display neurological abnormalities. We characterized the neuronal phenotype that results from Ofd1 inactivation in early phases of mouse embryonic development and at post-natal stages. We determined that Ofd1 plays a crucial role in forebrain development, and in particular, in the control of dorso-ventral patterning and early corticogenesis. We observed abnormal activation of Sonic hedgehog (Shh), a major pathway modulating brain development. Ultrastructural studies demonstrated that early Ofd1 inactivation results in the absence of ciliary axonemes despite the presence of mature basal bodies that are correctly orientated and docked. Ofd1 inducible-mediated inactivation at birth does not affect ciliogenesis in the cortex, suggesting a developmental stage-dependent role for a basal body protein in ciliogenesis. Moreover, we showed defects in cytoskeletal organization and apical-basal polarity in Ofd1 mutant embryos, most likely due to lack of ciliary axonemes. Thus, the present study identifies Ofd1 as a developmental disease gene that is critical for forebrain development and ciliogenesis in embryonic life, and indicates that Ofd1 functions after docking and before elaboration of the axoneme in vivo
Environmental concentrations of a delorazepam-based drug impact on embryonic development of non-target Xenopus laevis
Benzodiazepines, psychotropics drugs used for treating sleep disorders, anxiety and epilepsy, represent a major class of emerging water pollutants. As occurs for other pharmaceutical residues, they are not efficiently degraded during sewage treatment and persist in effluent waters. Bioaccumulation is already reported in fish and small crustaceans, but the impact and consequences on other “non-target” aquatic species are still unclear and nowadays of great interest. In this study, we investigated the effects of a pharmaceutical preparation containing the benzodiazepine delorazepam on the embryogenesis of Xenopus laevis, amphibian model species, taxa at high risk of exposure to water contaminants. Environmental (1 μg/L) and two higher (5 and 10 μg/L) concentrations were tested on tadpoles up to stage 45/46. Results demonstrate that delorazepam interferes with embryo development and that the effects are prevalently dose-dependent. Delorazepam reduces vitality by decreasing heart rate and motility, induces marked cephalic and abdominal edema, as well as intestinal and retinal defects. At the molecular level, delorazepam increases ROS production, modifies the expression of some master developmental genes and pro-inflammatory cytokines. The resulting stress condition significantly affects embryos’ development and threatens their survival. Similar effects should be expected as well in embryos belonging to other aquatic species that have not been yet considered targets for these pharmaceutical residues
Transient Anomalous Diffusion MRI in Excised Mouse Spinal Cord: Comparison Among Different Diffusion Metrics and Validation With Histology
Neural tissue is a hierarchical multiscale system with intracellular and extracellular
diffusion compartments at different length scales. The normal diffusion of bulk water
in tissues is not able to detect the specific features of a complex system, providing
nonlocal, diffusion measurement averaged on a 10-20 mm length scale. Being able to
probe tissues with sub-micrometric diffusion length and quantify new local parameters,
transient anomalous diffusion (tAD) would dramatically increase the diagnostic potential
of diffusion MRI (DMRI) in detecting collective and sub-micro architectural changes
of human tissues due to pathological damage. In DMRI, the use of tAD parameters
quantified using specific DMRI acquisition protocols and their interpretation has often
aroused skepticism. Although the derived formulas may accurately fit experimental
diffusion-weighted data, the relationships between the postulated dynamical feature
and the underlying geometrical structure remains elusive, or at most only suggestive.
This work aimed to elucidate and validate the image contrast and information
that can be obtained using the tAD model in white matter (WM) through a direct
comparison between different diffusion metrics and histology. Towards this goal,
we compared tAD metrics extracted from pure subdiffusion (a-imaging) and superpseudodiffusion (g-imaging) in excised mouse spinal cord WM, together with T2 and T2  relaxometry, conventional (normal diffusion-based) diffusion tensor imaging (DTI) and q-space imaging (QSI), with morphologic measures obtained by optical microscopy, to determine which structural and topological characteristics of myelinated axons influenced tAD contrast. Axon diameter (AxDiam), the standard deviation of diameters (SDax:diam), axonal density (AxDens) and effective local density (ELD) were extracted from optical images in several WM tracts. Among all the diffusion parameters obtained at 9.4 T, g-metrics confirmed a strong dependence on magnetic in-homogeneities quantified by R2  = 1/T2  and showed the strongest associations with AxDiam and ELD. On the other hand, a-metrics showed strong associations with SDax:diam and was significantly related to AxDens, suggesting its ability to quantify local heterogeneity degree in neural tissue. These results elucidate the biophysical mechanism underpinning tAD parameters and show the clinical potential of tAD-imaging, considering that both physiologic and pathologic neurodegeneration translate into alterations of WM morphometry and topology
Effects of Cadmium Exposure on Gut Villi in Danio rerio.
In aquatic organisms, cadmium exposure occurs from ovum to death and the route of
absorption is particularly wide, being represented by skin, gills and gastrointestinal tract, through
which contaminated water and/or preys are ingested. It is known that cadmium interferes with
the gut; however, less information is available on cadmium effects on an important component of
the gut, namely goblet cells, specialized in mucus synthesis. In the present work, we studied the
effects of two sublethal cadmium concentrations on the gut mucosa of Danio rerio. Particular attention
was paid to changes in the distribution of glycan residues, and in metallothionein expression in
intestinal cells. The results show that cadmium interferes with gut mucosa and goblet cells features.
The effects are dose- and site-dependent, the anterior gut being more markedly affected than the
midgut. Cadmium modifies the presence and/or distribution of glycans in the brush border and
cytoplasm of enterocytes and in the goblet cells’ cytoplasm and alters the metallothionein expression
and localization. The results suggest a significant interference of cadmium with mucosal efficiency,
representing a health risk for the organism in direct contact with contamination and indirectly for the
trophic chain
Seeing through the skin: dermal light sensitivity provides cryptism in moorish gecko
Concealment by means of colour change is a pre-eminent deceptive mechanism used by both predators and prey. The moorish gecko Tarentola mauritanica is able to blend into the background by either darkening or paling according to the substrate darkness. Here we examined the functioning of background perception in moorish gecko. We experimentally excluded the involvement of melanophore-stimulating hormone in camouflage. Blindfolded individuals change their colour consistently with the background. Surprisingly, individuals with covered flanks were not able to change colour, no matter whether they were allowed to see the substrate or not. Accordingly, we found high levels of opsin transcript and protein in the flank region of the gecko. These observations suggest that T.mauritanica skin melanophores are able to activate a process of colour change autonomously. This study yields the first evidence of crypsis mediated by dermal light sensitivity in amniote
Commercial Red Food Dyes Preparations Modulate the Oxidative State in Three Model Organisms (Cucumis sativus, Artemia salina, and Danio rerio)
The growing environmental spreading of food synthetic dyes and bio-colors have the
potential for altering organisms’ redox states. Here, three model species for aquatic pollution trials,
Cucumis sativus seeds, Artemia salina cysts, and Danio rerio embryos, were short-term exposed to
a fixed concentration of the artificial red E124, and two red bio-colors, cochineal E120, and vegan
red (VEGR). In the animal models, we evaluated the total reactive oxygen species (ROS) and the
susceptibility to in vitro oxidative stress, and in C. sativus, H2O2 production and antioxidant capacity.
We also measured organismal performance indices (routine oxygen consumption in the animal
models, dark oxygen consumption, and photosynthetic efficiency in C. sativus). In C. sativus, only
E124 increased ROS and affected dark oxygen consumption and photosynthetic efficiency, while
all dyes enhanced the antioxidant defenses. In the A. salina nauplii, all dyes increased ROS, while
E120 and E124 reduced the susceptibility to oxidative stress. In D. rerio, treatments did not affect
ROS content, and reduced oxidative stress susceptibility. Our data show that red food dyes affect
the redox state of the developing organisms, in which ROS plays a significant role. We suggest a
potentially toxic role for red food dyes with environmentally relevant consequences
Novel Approach for Evaluation of Bacteroides fragilis Protective Role against Bartonella henselae Liver Damage in Immunocompromised Murine Model
Bartonella henselae is a gram-negative facultative intracellular bacterium and is the causative agent of cat-scratch disease. Our previous data have established that Bacteroides fragilis colonization is able to prevent B. henselae damages through the polysaccharide A (PSA) in an experimental murine model. In order to determine whether the PSA is essential for the protection against pathogenic effects of B. henselae in immunocompromised hosts, SCID mice were co-infected with B. fragilis wild type or its mutant B. fragilis 1PSA and the effects of infection on murine tissues have been observed by High-Frequency Ultrasound (HFUS), histopathological examination, and Transmission Electron Microscopy (TEM). For the first time, echostructure, hepatic lobes length, vascular alterations, and indirect signs of hepatic dysfunctions, routinely used as signs of disease in humans, have been analyzed in an immunocompromised murine model. Our findings showed echostructural alterations in all infected mice compared with the Phosphate Buffer Solution (PBS) control group; further, those infected with B. henselae and co-infected with B. henselae/B. fragilis 1PSA presented the major echostructural alterations. Half of the mice infected with B. henselae and all those co-infected with B. henselae/B. fragilis 1PSA have showed an altered hepatic echogenicity compared with the renal cortex. The echogenicity score of co-infected mice with B. henselae/B. fragilis 1PSA differed significantly compared with the PBS control group (p < 0.05). Moreover the inflammation score of the histopathological evaluation was fairly concordant with ultrasound findings. Ultrastructural analysis performed by TEM revealed no significant alterations in liver samples of SCID mice infected with B. fragilis wild type while those infected with B. fragilis 1PSA showed the presence of collagen around the main vessels compared with the PBS control group. The liver samples of mice infected with B. henselae showed macro-areas rich in collagen, stellate cells, and histiocytic cells. Interestingly, our data demonstrated that immunocompromised SCID mice infected with B. henselaeand co-infected with B. henselae/B. fragilis ΔPSA showed the most severe morpho-structural liver damage. In addition, these results suggests that the HFUS together with histopathological evaluation could be considered good imaging approach to evaluate hepatic alterations
The Hepatic Mitochondrial Alterations Exacerbate Meta-Inflammation in Autism Spectrum Disorders
The role of the liver in autism spectrum disorders (ASD), developmental disabilities characterized by impairments in social interactions and repetitive behavioral patterns, has been poorly investigated. In ASD, it has been shown a dysregulation of gut-brain crosstalk, a communication system able to influence metabolic homeostasis, as well as brain development, mood and cognitive functions. The liver, with its key role in inflammatory and metabolic states, represents the crucial metabolic organ in this crosstalk. Indeed, through the portal vein, the liver receives not only nutrients but also numerous factors derived from the gut and visceral adipose tissue, which modulate metabolism and hepatic mitochondrial functions. Here, we investigated, in an animal model of ASD (BTBR mice), the involvement of hepatic mitochondria in the regulation of inflammatory state and liver damage. We observed increased inflammation and oxidative stress linked to hepatic mitochondrial dysfunction, steatotic hepatocytes, and marked mitochondrial fission in BTBR mice. Our preliminary study provides a better understanding of the pathophysiology of ASD and could open the way to identifying hepatic mitochondria as targets for innovative therapeutic strategies for the disease
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