Incontinentia pigmenti: a case associated with cardiovascular anomalies

Various cutaneous and developmental defects of the eyes, teeth, skeleton, and central nervous system have been detected in infants with incontinentia pigmenti. We report an isolated case of incontinentia pigmenti in a 6-month-old girl in association with tricuspid insufficiency, an abnormal shunt of the right pulmonary vein into the superior vena cava, and pulmonary hypertension. We believe that our findings will help to confirm the association of cardiovascular anomalies in IP.     

The combined effect of interleukin (IL)-10 and IL-12 polymorphisms on induced cytokine production

Interleukin (IL)-12 and IL-10 are immunoregulatory cytokines with an antagonistic effect of the T-helper (Th)1/Th2 cytokine balance and provide a functional link between innate and adaptive immune responses. The aim of the study was to investigate the combined effect of -1082A*G in IL10 and +16974A*C in IL12B single nucleotide polymorphisms (SNPs) on induced cytokine production by stimulated peripheral blood mononuclear cells (PBMCs) isolated from healthy donors. The presence of the high-producer IL-12p40 genotype led to diminished production of IL-10 as determined by the -1082*G allele of SNP in IL10. Significantly decreased IL-10 production was detected in AA+AG/GG in comparison with the low-producer IL-12p40 (AC/CC+AG/GG) genotype combination after stimulation with C3bgp (2 +/- 4 vs. 29 +/- 14.2 pg/ml; p = 0.0003) and LPS (33.4 +/- 13.5 vs. 93.3 +/- 59.6 pg/ml; p = 0.019). IL-12p40 production was independent of IL10 genotype. The present results demonstrated that the production of IL-10 from PBMC depended on both -1082A*G in IL10 and +16974A*C in IL12B polymorphisms.     

Translational bypass of nonsense mutations in zebrafish rep1, pax2.1 and lamb1 highlights a viable therapeutic option for untreatable genetic eye disease

The extensive molecular genetic heterogeneity seen with inherited eye disease is a major barrier to the development of gene-based therapeutics. The underlying molecular pathology in a considerable proportion of these diseases however are nonsense mutations leading to premature termination codons. A therapeutic intervention targeted at this abnormality would therefore potentially be relevant to a wide range of inherited eye diseases. We have taken advantage of the ability of aminoglycoside drugs to suppress such nonsense mutations and partially restore full-length, functional protein in a zebrafish model of choroideraemia (chm(ru848); juvenile chorio-retinal degeneration) and in two models of ocular coloboma (noi(tu29a) and gup(m189); congenital optic fissure closure defects). In vitro cell-based assays showed significant readthrough with two drugs, gentamicin and paromomycin, which was confirmed by western blot and in vitro prenylation assays. The presence of either aminoglycoside during zebrafish development in vivo showed remarkable prevention of mutant ocular phenotypes in each model and a reduction in multisystemic defects leading to a 1.5-1.7-fold increase in survival. We also identified a significant reduction in abnormal cell death shown by TUNEL assay. To test the hypothesis that optic fissure closure was apoptosis-dependent, the anti-apoptotic agents, curcumin and zVAD-fmk, were tested in gup(m189) embryos. Both drugs were found to reduce the size of the coloboma, providing molecular evidence that cell death is required for optic fissure remodelling. These findings draw attention to the value of zebrafish models of eye disease as useful preclinical drug screening tools in studies to identify molecular mechanisms amenable to therapeutic intervention.     

Viscoelastic Characteristics of <Emphasis Type="Italic">In Vitro</Emphasis> Vital and Devitalized Rat Aorta and Human Arterial Prostheses

The arterial wall viscoelasticity plays an essential role in the vascular responsiveness to vasoactive drugs or pathologies. The aim of this investigation was to derive and compare resonance curve (RC), natural frequency (f ₀), dynamic modulus of elasticity (E′), and coefficient of viscosity (β) of (i) vital and devitalized preparations of rat thoracic and abdominal aorta, (ii) human arterial prostheses, and to study the histomorphology of vital and devitalized rat aorta. The method of low frequency forced oscillations was employed. RC of vital preparations showed a hardening type of elasticity whereas in devitalized preparations it was of softening type. E′ increased nonlinearly, f ₀ decreased and β increased linearly with equivalent intraluminal pressure (p _eqi). Distensibility of abdominal aorta was lower than thoracic aorta. Distensibility decreased with increasing p _eqi. E′, f ₀, and β increased significantly after devitalization. It was suggested that postmortem viscoelastic characteristics should not be used directly to specify the vital arteries viscoelasticity. RC of human prostheses showed a softening type of elasticity. Arterial prostheses have low circumferential distensibility with E′-values higher than reported in the literature for human arteries. The method of forced oscillations could be employed for studying the arterial wall biomechanics and viscoelasticity of arterial prostheses. The experiments were carried out in Laboratory Integrative Neuropharmacology, Institute of Neurobiology, BAS, Sofia, Bulgaria and the Department of Anatomy and Histology, Medical University Varna, Varna, Bulgaria.     

Automatic Segmentation of MRI Images in Dynamic Programming Mode

Objective: Purpose of this work was to develop methods contour and volume of areas of interest definition intomographic images of the breast. Methods: The study included images of the breast of 13 patients obtained on an openelectronic resource The Breast-MRI-NACT-Pilot image collection. Statistical processing of the data was carried out, thereliability of the results of calculating the volumes of the breast areas was estimated, a visual evaluation of the obtainednumerical values was provided – a linear graph. Result: A program for automatic determination of breast volume andvolume of pathology has been developed and tested. For segmenting areas of the breast, a threshold segmentation anda managed watershed method programs were written in Matlab package. The developed programs allowed to obtainreliable data when processing MRI images of the breast of 13 patients. Results of using Hurst exponent show that inthe case of a pathology, the exponent is less than 0.4, and for the tissue without pathology the Hurst index is greaterthan 0.4. This method is implemented in dynamic programming mode, which allows to adjust the algorithm for a taskof examining images. Conclusion: The developed methods of definition of contours and calculating volumes allow anautomatic quantitative evaluation of the ratio of the volumes of different identified areas of interest in the postprocessingof MRI images. Also, the results have established that it is possible to use the Hurst exponent as an additional tool foridentifying pathologies in areas of interest.     

Staurosporine-induced differentiation of the RGC-5 cell line leads to apoptosis and cell death at the lowest differentiating concentration

Background

Supplementation of staurosporine is the method of choice for differentiating the solely existing retinal ganglion cell (RGC)-5 cell line. This differentiation was initially claimed to be in the absence of apoptosis, but some publications supposed the induction of apoptosis during staurosporine induced RGC-5 differentiation. In respect to these inconsistencies in the literature, we investigated in detail whether RGC-5 cell differentiation by staurosporine induces apoptosis or not.

Methods

Amounts of 50?nM, 200?nM, 300?nM, and 600?nM of staurosporine were supplemented on RGC-5 cells for 24 h. Cell morphology and cell death, via propidium iodide staining, were evaluated with phase contrast and fluorescence microscopy, respectively. Cell amount, cell proliferation, and cell viability were analyzed by crystal violet staining, CFSE flow cytometry, and MTS assay, respectively. Apoptosis was determined by analyzing caspase 3/7 activity, Annexin-V+/ 7AAD- cells and the quotient of Bax to Bcl-2 mRNA expression via caspase 3/7 activity assay, flow cytometry, and real-time PCR, respectively.

Results

RGC-5 cells started to change their morphology and their expression of neuronal markers at 50?nM of staurosporine. This was associated with apoptosis and cell death, as indicated by a 2.1-fold (p?<?0.0005) increase in caspase 3/7 activity, a 1.2–fold (p?<?0.05) induction of Annexin-V+/ 7AAD- cells, and a 12–fold (p?<?0.0005) increase in propidium iodide positive cells, respectively. Furthermore, staurosporine led to a dose-dependent increase in apoptosis and reduction in cell viability, cell density, and cell proliferation.

Conclusions

The lowest staurosporine concentration inducing RGC-5 cell differentiation is accompanied by apoptosis and cell death.     

Enhancing the Immunogenicity of Vaccinia Virus

The conventional live smallpox vaccine based on the vaccinia virus (VACV) cannot be widely used today because it is highly reactogenic. Therefore, there is a demand for designing VACV variants possessing enhanced immunogenicity, making it possible to reduce the vaccine dose and, therefore, significantly eliminate the pathogenic effect of the VACV on the body. In this study, we analyzed the development of the humoral and T cell-mediated immune responses elicited by immunizing mice with low-dose VACV variants carrying the mutant A34R gene (which increases production of extracellular virions) or the deleted A35R gene (whose protein product inhibits antigen presentation by the major histocompatibility complex class II). The VACV LIVP strain, which is used as a smallpox vaccine in Russia, and its recombinant variants LIVP-A34R*, LIVP-dA35R, and LIVP-A34R*-dA35R, were compared upon intradermal immunization of BALB/c mice at a dose of 10⁴ pfu/animal. The strongest T cell-mediated immunity was detected in mice infected with the LIVP-A34R*-dA35R virus. The parental LIVP strain induced a significantly lower antibody level compared to the strains carrying the modified A34R and A35R genes. Simultaneous modification of the A34R gene and deletion of the A35R gene in VACV LIVP synergistically enhanced the immunogenic properties of the LIVP-A34R*-dA35R virus.     

Female reproductive tract microbiome and early miscarriages

Miscarriage is one of the main causes of reproductive loss, which can lead to a number of physical and psychological complications and other long-term consequences. However, the role of vaginal and uterine microbiome in such complications is poorly understood. To review the published data on the function of the female reproductive tract microbiome in the pathogenesis of early miscarriages. The articles published over the past 20 years and deposited in PubMed, Google Academy, Scopus, Elibrary, ResearchGate, and EBSCO databases were analyzed. The review presents new data on the impact of the vaginal and uterine microbiome on the local immunity, including defense against sexually transmitted infections, and its association with other factors of miscarriages. The studies on the microbiome of non-pregnant women with recurrent miscarriages in the anamnesis, patients undergoing IVF, and pregnant women with miscarriages, as well as new directions in the microbiome research are discussed. The majority of studies have demonstrated that the dominant species of the vaginal and uterine microbiome in patients with early miscarriages are non-Lactobacillus bacteria. As many of these bacteria have not previously been detected by cultural studies and their role in obstetric complications is not well defined, further research on the female reproductive tract microbiome, including the microbiome of the cervix uteri, is needed to develop new approaches for the prognosis and prevention of miscarriages.     

Review: Digital experiences and their impact on the lives of adolescents with pre-existing anxiety,depression, eating and nonsuicidal self-injury conditions – a systematic review

Background

Published systematic reviews provide evidence linking positive and negative digital experiences to adolescent mental health. However, these reviews focus on the general public rather than the digital experiences of adolescents with different pre-existing mental health conditions and so may be limited in their clinical relevance. We review publications relating to anxiety, depression, eating disorders and nonsuicidal self-injury to identify common and condition-specific digital experiences and how these may be implicated in the origins and maintenance of these mental health conditions.

Methods

A systematic literature search using a combination of mental health, digital experience (including social media use), and age of the target population terms was conducted on four databases. Detailed findings from the included studies were summarised using a combination of thematic and narrative methods.

Results

Five qualitative and 21 quantitative studies met the eligibility criteria for inclusion (n = 5021). Nine studies included adolescents with depression, one with eating problems, two with nonsuicidal self-injury and 14 with multiple emotional health conditions. The review identified six themes related to the target populations' digital experiences: (a) social connectivity and peer support; (b) escape and/or distraction; (c) social validation and social comparison; (d) accessing/creation of potentially harmful content; (e) cyberbullying; and (f) difficulties with self-regulation during engagement with digital media.

Conclusions

Digital practices of adolescents with pre-existing clinical vulnerabilities are complex and encompass a range of positive and negative experiences, which appear to have common elements across different clinical populations. The literature is currently too limited to identify disorder-specific practices, with too few direct or indirect comparisons between conditions.