The chromosomal passenger complex and the spindle assembly checkpoint: kinetochore-microtubule error correction and beyond

During mitosis, correct bipolar chromosome attachment to the mitotic spindle is an essential prerequisite for the equal segregation of chromosomes. The spindle assembly checkpoint can prevent chromosome segregation as long as not all chromosome pairs have obtained bipolar attachment to the spindle. The chromosomal passenger complex plays a crucial role during chromosome alignment by correcting faulty chromosome-spindle interactions (e.g. attachments that do not generate tension). In the process of doing so, the chromosomal passenger complex generates unattached chromosomes, a specific situation that is known to promote checkpoint activity. However, several studies have implicated an additional, more direct role for the chromosomal passenger complex in enforcing the mitotic arrest imposed by the spindle assembly checkpoint. In this review, we discuss the different roles played by the chromosomal passenger complex in ensuring proper mitotic checkpoint function. Additionally, we discuss the possibility that besides monitoring the presence of unattached kinetochores, the spindle assembly checkpoint may also be capable of responding to chromosome-microtubule interactions that do not generate tension and we propose experimental set-ups to study this

Hyperoxia Causes Mitochondrial Fragmentation in Pulmonary Endothelial Cells by Increasing Expression of Pro-Fission Proteins

Objective—We explored mechanisms that alter mitochondrial structure and function in pulmonary endothelial cells (PEC) function after hyperoxia. Approach and Results—Mitochondrial structures of PECs exposed to hyperoxia or normoxia were visualized and mitochondrial fragmentation quantified. Expression of pro-fission or fusion proteins or autophagy-related proteins were assessed by Western blot. Mitochondrial oxidative state was determined using mito-roGFP. Tetramethylrhodamine methyl ester estimated mitochondrial polarization in treatment groups. The role of mitochondrially derived reactive oxygen species in mt-fragmentation was investigated with mito-TEMPOL and mitochondrial DNA (mtDNA) damage studied by using ENDO III (mt-tat-endonuclease III), a protein that repairs mDNA damage. Drp-1 (dynamin-related protein 1) was overexpressed or silenced to test the role of this protein in cell survival or transwell resistance. Hyperoxia increased fragmentation of PEC mitochondria in a time-dependent manner through 48 hours of exposure. Hyperoxic PECs exhibited increased phosphorylation of Drp-1 (serine 616), decreases in Mfn1 (mitofusion protein 1), but increases in OPA-1 (optic atrophy 1). Pro-autophagy proteins p62 (LC3 adapter–binding protein SQSTM1/p62), PINK-1 (PTEN-induced putative kinase 1), and LC3B (microtubule-associated protein 1A/1B-light chain 3) were increased. Returning cells to normoxia for 24 hours reversed the increased mt-fragmentation and changes in expression of pro-fission proteins. Hyperoxia-induced changes in mitochondrial structure or cell survival were mitigated by antioxidants mito-TEMPOL, Drp-1 silencing, or inhibition or protection by the mitochondrial endonuclease ENDO III. Hyperoxia induced oxidation and mitochondrial depolarization and impaired transwell resistance. Decrease in resistance was mitigated by mito-TEMPOL or ENDO III and reproduced by overexpression of Drp-1. Conclusions—Because hyperoxia evoked mt-fragmentation, cell survival and transwell resistance are prevented by ENDO III and mito-TEMPOL and Drp-1 silencing, and these data link hyperoxia-induced mt-DNA damage, Drp-1 expression, mt-fragmentation, and PEC dysfunction

Missense MED12 variants in 22 males with intellectual disability: From nonspecific symptoms to complete syndromes

Genotype; Intellectual disability; PhenotypeGenotipo; Discapacidad intelectual; FenotipoGenotip; Discapacitat intel·lectual; FenotipWe describe the phenotype of 22 male patients (20 probands) carrying a hemizygous missense variant in MED12. The phenotypic spectrum is very broad ranging from nonspecific intellectual disability (ID) to the three well-known syndromes: Opitz–Kaveggia syndrome, Lujan–Fryns syndrome, or Ohdo syndrome. The identified variants were randomly distributed throughout the gene (p = 0.993, χ2 test), but mostly outside the functional domains (p = 0.004; χ2 test). Statistical analyses did not show a correlation between the MED12-related phenotypes and the locations of the variants (p = 0.295; Pearson correlation), nor the protein domain involved (p = 0.422; Pearson correlation). In conclusion, establishing a genotype–phenotype correlation in MED12-related diseases remains challenging. Therefore, we think that patients with a causative MED12 variant are currently underdiagnosed due to the broad patients' clinical presentations.Foundation for Science and Technology (FCT), Grant/Award Number: UIDB/00215/2020 UIDP/00215/2020 LA/P/0064/2020; Broad Institute; National Eye Institute; National Heart, Lung and Blood Institute, Grant/Award Number: UM1 HG008900; National Human Genome Research Institute, Grant/Award Number: R01 HG009141

Fluctuations, dissipation and the dynamical Casimir effect

Vacuum fluctuations provide a fundamental source of dissipation for systems coupled to quantum fields by radiation pressure. In the dynamical Casimir effect, accelerating neutral bodies in free space give rise to the emission of real photons while experiencing a damping force which plays the role of a radiation reaction force. Analog models where non-stationary conditions for the electromagnetic field simulate the presence of moving plates are currently under experimental investigation. A dissipative force might also appear in the case of uniform relative motion between two bodies, thus leading to a new kind of friction mechanism without mechanical contact. In this paper, we review recent advances on the dynamical Casimir and non-contact friction effects, highlighting their common physical origin.Comment: 39 pages, 4 figures. Review paper to appear in Lecture Notes in Physics, Volume on Casimir Physics, edited by Diego Dalvit, Peter Milonni, David Roberts, and Felipe da Rosa. Minor changes, a reference adde

A DNA Vaccine against Yellow Fever Virus: Development and Evaluation

Attenuated yellow fever (YF) virus 17D/17DD vaccines are the only available protection from YF infection, which remains a significant source of morbidity and mortality in the tropical areas of the world. The attenuated YF virus vaccine, which is used worldwide, generates both long-lasting neutralizing antibodies and strong T-cell responses. However, on rare occasions, this vaccine has toxic side effects that can be fatal. This study presents the design of two non-viral DNA-based antigen formulations and the characterization of their expression and immunological properties. The two antigen formulations consist of DNA encoding the full-length envelope protein (p/YFE) or the full-length envelope protein fused to the lysosomal-associated membrane protein signal, LAMP-1 (pL/YFE), aimed at diverting antigen processing/presentation through the major histocompatibility complex II precursor compartments. The immune responses triggered by these formulations were evaluated in H2b and H2d backgrounds, corresponding to the C57Bl/6 and BALB/c mice strains, respectively. Both DNA constructs were able to induce very strong T-cell responses of similar magnitude against almost all epitopes that are also generated by the YF 17DD vaccine. The pL/YFE formulation performed best overall. In addition to the T-cell response, it was also able to stimulate high titers of anti-YF neutralizing antibodies comparable to the levels elicited by the 17DD vaccine. More importantly, the pL/YFE vaccine conferred 100% protection against the YF virus in intracerebrally challenged mice. These results indicate that pL/YFE DNA is an excellent vaccine candidate and should be considered for further developmental studies

Doctor, how much weight will I lose? - a new individualized predictive model for weight loss

Bariatric surgery is an effective treatment for weight loss, but the patient’s ability to reach a sustained weight loss depends upon several technical and individual factors. Creating an easy model that adapts bariatric surgery’s weight loss goals for each patient is very important for pre-surgery and follow-up evaluations.info:eu-repo/semantics/publishedVersio

Eyespot resistance gene Pch-1 from Aegilops ventricosa is associated with a different chromosome in wheat line H-93-70 than the resistance factor in "Roazon" wheat

The hexaploid wheat line H-93-70 carries a gene (Pch-1) that has been transferred from the wild grass Aegilops ventricosa and confers a high degree of resistance to eyespot diesease, caused by the fungus Pseudocercosporella herpotrichoides. Crosses of the resistant line H-93-70 with the susceptible wheat Pané 247 and with a 7D/7Ag wheat/Agropyron substitution line were carried out and F2 kernels were obtained. The kernels were cut transversally and the halves carrying the embryos were used for the resistance test, while the distal halves were used for genetic typing. Biochemical markers were used to discriminate whether the transferred Pch-1 gene was located in chromosome 7D, as is the case for a resistance factor present in Roazon wheat. In the crosses involving Pané 247, resistance was not associated with the 7D locus Pln, which determines sterol ester pattern (dominant allele in H-93-70). In the crosses with the 7D/7Ag substitution line, resistance was neither associated with protein NGE-11 (7D marker), nor alternatively inherited with respect to protein C-7 (7Ag marker). It is concluded that gene Pch-1 represents a different locus and is not an allele of the resistance factor in Roazon whea

Cytological analysis of hybrids among triticales and trigopiros

We studied three different tricepiros: (Don Santiago x Don Noé), (Cumé x Horovitz) and (Cumé x Don Noé). The tricepiro (Don Santiago x Don Noé) was obtained by crossing the triticale Don Santiago INTA (AABBRR, 2n = 6x = 42) with the trigopiro Don Noé INTA (AABBDDJJ, 2n = 8x = 56). The number of chromosomes for the F1 was 2n = 49, the most frequent meiotic configuration being 14 bivalents and 21 univalents. The univalents were situated in the periphery of the equatorial plane, whereas the bivalents were located in the central zone. The chromatids in some of the univalents split when bivalents underwent reductional division in anaphase I. There were few laggard chromosomes or chromatids at this phase. The number of chromosomes (2n = 48-58) was high and variable, and the number of bivalents per cell (18-23) also high in F 3 individuals. In all F 8 tricepiros (Don Santiago x Don Noé), F 12 tricepiros (Cumé x Horovitz) and F 12 tricepiros (Cumé x Don Noé), the number of chromosomes (2n = 42) was the same, these retaining the rye genome, as demonstrated by GISH and FISH. These new synthesized allopolyploids constitute interesting models for investigating the evolutionary changes responsible for diploidization, and the chromosomal and genomic re-ordering that cannot be revealed in natural allopolyploids

Transcription of toll-like receptors 2, 3, 4 and 9, FoxP3 and Th17 cytokines in a susceptible experimental model of canine Leishmania infantum infection

Canine leishmaniosis (CanL) due to Leishmania infantum is a chronic zoonotic systemic disease resulting from complex interactions between protozoa and the canine immune system. Toll-like receptors (TLRs) are essential components of the innate immune system and facilitate the early detection of many infections. However, the role of TLRs in CanL remains unknown and information describing TLR transcription during infection is extremely scarce. The aim of this research project was to investigate the impact of L. infantum infection on canine TLR transcription using a susceptible model. The objectives of this study were to evaluate transcription of TLRs 2, 3, 4 and 9 by means of quantitative reverse transcription polymerase chain reaction (qRT-PCR) in skin, spleen, lymph node and liver in the presence or absence of experimental L. infantum infection in Beagle dogs. These findings were compared with clinical and serological data, parasite densities in infected tissues and transcription of IL-17, IL-22 and FoxP3 in different tissues in non-infected dogs (n = 10), and at six months (n = 24) and 15 months (n = 7) post infection. Results revealed significant down regulation of transcription with disease progression in lymph node samples for TLR3, TLR4, TLR9, IL-17, IL-22 and FoxP3. In spleen samples, significant down regulation of transcription was seen in TLR4 and IL-22 when both infected groups were compared with controls. In liver samples, down regulation of transcription was evident with disease progression for IL-22. In the skin, upregulation was seen only for TLR9 and FoxP3 in the early stages of infection. Subtle changes or down regulation in TLR transcription, Th17 cytokines and FoxP3 are indicative of the silent establishment of infection that Leishmania is renowned for. These observations provide new insights about TLR transcription, Th17 cytokines and Foxp3 in the liver, spleen, lymph node and skin in CanL and highlight possible markers of disease susceptibility in this model