ULP-2 SUMO protease regulates UPRmt and mitochondrial homeostasis in Caenorhabditis elegans.

Mitochondria are the powerhouses of cells, responsible for energy production and regulation of cellular homeostasis. When mitochondrial function is impaired, a stress response termed mitochondrial unfolded protein response (UPRmt) is initiated to restore mitochondrial function. Since mitochondria and UPRmt are implicated in many diseases, it is important to understand UPRmt regulation. In this study, we show that the SUMO protease ULP-2 has a key role in regulating mitochondrial function and UPRmt. Specifically, down-regulation of ulp-2 suppresses UPRmt and reduces mitochondrial membrane potential without significantly affecting cellular ROS. Mitochondrial networks are expanded in ulp-2 null mutants with larger mitochondrial area and increased branching. Moreover, the amount of mitochondrial DNA is increased in ulp-2 mutants. Downregulation of ULP-2 also leads to alterations in expression levels of mitochondrial genes involved in protein import and mtDNA replication, however, mitophagy remains unaltered. In summary, this study demonstrates that ULP-2 is required for mitochondrial homeostasis and the UPRmt.

Dynein directs prophase centrosome migration to control the stem cell division axis in the developing Caenorhabditis elegans epidermis.

The microtubule motor dynein is critical for the assembly and positioning of mitotic spindles. In Caenorhabditis elegans, these dynein functions have been extensively studied in the early embryo but remain poorly explored in other developmental contexts. Here, we use a hypomorphic dynein mutant to investigate the motor's contribution to asymmetric stem cell-like divisions in the larval epidermis. Live imaging of seam cell divisions that precede formation of the seam syncytium shows that mutant cells properly assemble but frequently misorient their spindle. Misoriented divisions misplace daughter cells from the seam cell row, generate anucleate compartments due to aberrant cytokinesis, and disrupt asymmetric cell fate inheritance. Consequently, the seam becomes disorganized and populated with extra cells that have lost seam identity, leading to fatal epidermal rupture. We show that dynein orients the spindle through the cortical GOA-1Gα-LIN-5NuMA pathway by directing the migration of prophase centrosomes along the anterior-posterior axis. Spindle misorientation in the dynein mutant can be partially rescued by elongating cells, implying that dynein-dependent force generation and cell shape jointly promote correct asymmetric division of epithelial stem cells.

Measurement of Shear Wave Speed and Normalized Elastic Modulus of Human Skin with and without Dermal Striae Using Shear Wave Elastography.

Supersonic shear imaging is a non-invasive technique used for detecting physiologic and pathologic changes in biological tissues. In this study, supersonic shear imaging was used to measure and compare shear wave speed (cs) and normalized elastic modulus (EN) values of skin with and skin without dermal striae (DS) in vivo. The values were measured at angles of 0°, 45°, 90° and 315° to the skin tension lines. In the presence of DS, a statistically significant reduction in the elasticity dermis was observed (p value <0.05). The mean values of cs and EN for STLs were higher in normal skin at 45° (4.26 ± 1.05 m/s and 56.23 ± 25.31 kPa) and 90° (4.26 ± 0.55 m/s and 54.91 ± 14.22 kPa), and those for DS were also higher at 45° (3.59 ± 0.72 m/s and 42.71 ± 27.97 kPa) and 90° (3.52 ± 0.65 m/s and 42.34 ± 31.68 kPa) than at other angles. Supersonic shear imaging was found to be a promising technique in the study of skin with DS. The data obtained in this study are expected to be relevant for future studies using shear wave elastography for the aforementioned purpose.

Quality of life in chronic pain patients: Illness- and wellness-focused coping as moderators.

Chronic pain is a health problem that significantly influences patients' lives, causing functional, social, socioeconomic, and emotional changes that impact quality of life (QoL). The aim of this study was to evaluate which variables (e.g., psychological morbidity, illness representations, pain, and coping) contribute to QoL and to analyse the moderating role of illness- and wellness-focused coping in the relationship between pain interference and Qol, in chronic pain patients. A sociodemographic and clinical questionnaire, the Brief Illness Perception Questionnaire, the Pain Catastrophizing Scale, the Hospital Anxiety and Depression Scale, the Chronic Pain Coping Inventory, the Short Form Health Survey, and the Brief Pain Inventory were completed by 103 patients with chronic pain. Greater use of wellness-focused coping and being professionally active were associated with better physical QoL. Cognitive representations and illness-focused coping contributed to physical QoL, and psychological morbidity contributed to mental QoL. Illness-focused coping and wellness-focused coping moderated the relationship between pain interference and physical QoL, but not with mental QoL. Since pain interference was positively related to psychological morbidity, and the latter was negatively related to QoL, it is extremely important to evaluate and promote patients' coping strategies that are focused on well-being to improve QoL. Results from this study underline the relevance of a multidisciplinary approach to chronic pain and the need to account for psychological morbidity and coping strategies in intervention programs to promote QoL in chronic pain patients.

A Path Toward Inclusive Social Cohesion: The Role of European and National Identity on Contesting vs. Accepting European Migration Policies in Portugal.

The Western hemisphere has witnessed recent increased immigration flows generating social and political debate across Europe. In one view, migration flows represent an opportunity to construct a diverse social cohesion. In another view, migration flows are perceived as a threat to existent national cultures. This view is held by political nationalisms and right-wing populist forces installed in the majority of EU countries' parliaments, accentuating discrimination against immigrants and residents in Europe. We theorize that European identity predicts positive attitudes toward immigrants (prosocial behavior and support for inclusive policies), whereas national identity's predictions of attitudes toward immigrants' inclusion depends on participants' political tendency. Moreover, we test the mediation effect of positive (humanitarian concerns and economic benefit) and negative (jobs scarcity, cultural deterioration, and invasion) arguments used in political discourses regarding immigrants' inclusion on the relation between national and European identities and attitudes toward immigrants' inclusion. Results (Portuguese sample, N = 176) show that national identity predicts negative attitudes toward immigrants' inclusion, but only among right-wing individuals. Among left-wing individuals, national identity predicts less contestation to immigrant's inclusion sustained by humanitarian concerns. Interestingly, European identification weakened right-wing individuals' adherence to discriminatory arguments and increased perceived economic contribution that immigrants bring to society, increasing agreement with prosocial behavior and immigrants' inclusion. We discuss that European identity, sustained in humanitarian values and economic benefit, may stimulate a stronger multicultural social cohesion, intergroup trust, and social well-being based on democratic values, social justice, and equality, and on the respect for human dignity.

Correction: Boronic acids as building blocks for the construction of therapeutically useful bioconjugates.

Correction for 'Boronic acids as building blocks for the construction of therapeutically useful bioconjugates' by João P. M. António et al., Chem. Soc. Rev., 2019, 48, 3513-3536, DOI: .

Cyanine-Like Boronic Acid-Derived Salicylidenehydrazone Complexes (Cy-BASHY) for Bioimaging Applications.

Boronic acid-derived salicylidenehydrazone complex (BASHY) dyes with a polymethine backbone were designed to yield efficient red-emitting and two-photon absorbing fluorophores that can be used as markers for astrocytes. The dyes are chemically stable in aqueous solution and do not undergo photodecomposition. Their photophysical properties can be electronically fine-tuned and thereby adapted to potentially different imaging situations and requirements.

Game of Tissues: How the Epidermis Thrones C. elegans Shape.

The versatility of epithelial cell structure is universally exploited by organisms in multiple contexts. Epithelial cells can establish diverse polarized axes within their tridimensional structure which enables them to flexibly communicate with their neighbors in a 360° range. Hence, these cells are central to multicellularity, and participate in diverse biological processes such as organismal development, growth or immune response and their misfunction ultimately impacts disease. During the development of an organism, the first task epidermal cells must complete is the formation of a continuous sheet, which initiates its own morphogenic process. In this review, we will focus on the C. elegans embryonic epithelial morphogenesis. We will describe how its formation, maturation, and spatial arrangements set the final shape of the nematode C. elegans. Special importance will be given to the tissue-tissue interactions, regulatory tissue-tissue feedback mechanisms and the players orchestrating the process.

Using high-resolution ultrasound imaging to characterize dermal striae in human skin.

BACKGROUND AND OBJECTIVE: The improvement in the appearance of the skin with dermal striae (DS) is currently eval-uated by invasive methods, such as biopsy. This study evaluates whether high-resolution ultrasound (HRUS) could be used to identify skin lesions in vivo caused by DS, using 2D images and measuring the thickness of the dermal layer. METHODS: High-resolution ultrasound at frequencies of 20 and 30 MHz was used in this study in ten volunteers with DS. The thickness of the skin layers was estimated by tracing five vertical lines from epidermis (EP) to dermis (DE) and DE to hypodermis (H) surface. RESULTS: The dermal lesions caused by striae appeared in ultrasonic images as poor echo areas. The average normal DE thickness varied from 1.07 to 1.65 mm, while the DE thickness with DS varied between 0.35 and 1.33 mm. A statistically significant reduction in the DE thickness was found (P-value < .05) in the presence of DS. The mean values of the EP thickness without and with DS were 0.12 ± 0.03 mm and 0.11 ± 0.02 mm, respec-tively. A total of 90.00% of the EP-related groups did not present the normal distribu-tion (P-value < .05). CONCLUSIONS: High-resolution ultrasound permitted the visualization of the three skin layers and the dermal lesions caused by striae. The dermal layer thicknesses with striae were thinner than those without. Therefore, ultrasound 2D imaging has shown to be a promising and financially feasible tool to be used as a noninvasive diagnostic method for evaluating therapeutic protocols used in the treatment of these dermal conditions.

The contribution of purpose in life to psychological morbidity and quality of life in chronic pain patients.

Chronic pain is a cause of morbidity, interference with daily functioning, decreased health and quality of life. Purpose in life acts as a protective factor and mitigates these consequences. This cross-sectional study aimed to determine whether purpose in life contributed to psychological morbidity and quality of life in patients with chronic pain by controlling psychological variables related to health (pain severity and interference, pain perceptions, pain catastrophizing and coping). The sample included 103 patients diagnosed with chronic pain. Results showed that purpose in life independently contributed to psychological morbidity and to mental quality of life, but not to physical quality of life, after controlling for pain-related variables. Results showed the relevance of purpose in life to identify patients at risk of developing psychological morbidity and decreased quality of life, suggestting the need to intervene in chronic pain, specifically on purpose in life, to prevent psychological morbidity and promote quality of life, in this population.

Boronic acids as building blocks for the construction of therapeutically useful bioconjugates.

Bioconjugates are multifunctional constructs in which biomolecules like peptides, proteins, vitamins and nucleic acids are endowed with the properties of specific payloads. These constructs recently emerged as a new generation of high-precision therapeutics, with several representatives reaching the market. This success stimulated an intense search for new biocompatible synthetic methodologies to connect both components and to control the bioconjugate's function. Despite the remarkable advances made in this field, most of the technologies developed for the construction of bioconjugates were engineered to yield stable constructs that can endure complex physiological conditions. Because of this, the use of reversible covalent bonds in the synthesis of bioconjugates has been rather overlooked, notwithstanding the potential of this strategy to generate stimuli responsive constructs that may operate in areas like the selective delivery of drugs, live-cell imaging and new theranostic approaches. Boronic acids are a well-known class of reagents that have been widely used in modern synthesis for the formation of C-C and C-heteroatom bonds. Apart from this, boronic acids exhibit an exquisite reversible coordination profile that can be explored as a molecular construction tool featuring specific mechanisms to control the structure and biological properties of bioconjugates. In this review, the use of boronic acids in the construction of therapeutically useful bioconjugates will be discussed, focusing on the molecular mechanisms that allow the use of these reagents as bioconjugation warheads, as central pieces of linker structures and as functional payloads.

A transient helix in the disordered region of dynein light intermediate chain links the motor to structurally diverse adaptors for cargo transport.

All animal cells use the motor cytoplasmic dynein 1 (dynein) to transport diverse cargo toward microtubule minus ends and to organize and position microtubule arrays such as the mitotic spindle. Cargo-specific adaptors engage with dynein to recruit and activate the motor, but the molecular mechanisms remain incompletely understood. Here, we use structural and dynamic nuclear magnetic resonance (NMR) analysis to demonstrate that the C-terminal region of human dynein light intermediate chain 1 (LIC1) is intrinsically disordered and contains two short conserved segments with helical propensity. NMR titration experiments reveal that the first helical segment (helix 1) constitutes the main interaction site for the adaptors Spindly (SPDL1), bicaudal D homolog 2 (BICD2), and Hook homolog 3 (HOOK3). In vitro binding assays show that helix 1, but not helix 2, is essential in both LIC1 and LIC2 for binding to SPDL1, BICD2, HOOK3, RAB-interacting lysosomal protein (RILP), RAB11 family-interacting protein 3 (RAB11FIP3), ninein (NIN), and trafficking kinesin-binding protein 1 (TRAK1). Helix 1 is sufficient to bind RILP, whereas other adaptors require additional segments preceding helix 1 for efficient binding. Point mutations in the C-terminal helix 1 of Caenorhabditis elegans LIC, introduced by genome editing, severely affect development, locomotion, and life span of the animal and disrupt the distribution and transport kinetics of membrane cargo in axons of mechanosensory neurons, identical to what is observed when the entire LIC C-terminal region is deleted. Deletion of the C-terminal helix 2 delays dynein-dependent spindle positioning in the one-cell embryo but overall does not significantly perturb dynein function. We conclude that helix 1 in the intrinsically disordered region of LIC provides a conserved link between dynein and structurally diverse cargo adaptor families that is critical for dynein function in vivo.

Five-Component Self-Assembly of Cucurbituril-Based Hetero-pseudorotaxanes.

[5]Pseudorotaxanes can be obtained by self-sorting using heteroditopic guests and various cucurbituril homologues as hosts. The assembly and chemically induced disassembly of the pseudorotaxanes can be monitored by measuring the fluorescence of the anthracene guest in solution. Mass spectral evidence for the supramolecular assemblies is obtained in the gas phase. The disassembly in the gas phase can be achieved by collision-induced dissociation leading to the corresponding [2]- and [3]pseudorotaxanes.

Molecular mechanism of dynein recruitment to kinetochores by the Rod-Zw10-Zwilch complex and Spindly.

The molecular motor dynein concentrates at the kinetochore region of mitotic chromosomes in animals to accelerate spindle microtubule capture and to control spindle checkpoint signaling. In this study, we describe the molecular mechanism used by the Rod-Zw10-Zwilch complex and the adaptor Spindly to recruit dynein to kinetochores in Caenorhabditis elegans embryos and human cells. We show that Rod's N-terminal ß-propeller and the associated Zwilch subunit bind Spindly's C-terminal domain, and we identify a specific Zwilch mutant that abrogates Spindly and dynein recruitment in vivo and Spindly binding to a Rod ß-propeller-Zwilch complex in vitro. Spindly's N-terminal coiled-coil uses distinct motifs to bind dynein light intermediate chain and the pointed-end complex of dynactin. Mutations in these motifs inhibit assembly of a dynein-dynactin-Spindly complex, and a null mutant of the dynactin pointed-end subunit p27 prevents kinetochore recruitment of dynein-dynactin without affecting other mitotic functions of the motor. Conservation of Spindly-like motifs in adaptors involved in intracellular transport suggests a common mechanism for linking dynein to cargo.

Site-selective installation of BASHY fluorescent dyes to Annexin V for targeted detection of apoptotic cells.

Fluorophores are indispensable for imaging biological processes. We report the design and synthesis of azide-tagged boronic acid salicylidenehydrazone (BASHY) dyes and their use for site-selective labelling of Annexin V. The Annexin V-BASHY conjugate maintained function and fluorescence as demonstrated by the targeted detection of apoptotic cells.

A Three-Component Assembly Promoted by Boronic Acids Delivers a Modular Fluorophore Platform (BASHY Dyes).

The modular assembly of boronic acids with Schiff-base ligands enabled the construction of innovative fluorescent dyes [boronic acid salicylidenehydrazone (BASHY)] with suitable structural and photophysical properties for live cell bioimaging applications. This reaction enabled the straightforward synthesis (yields up to 99%) of structurally diverse and photostable dyes that exhibit a polarity-sensitive green-to-yellow emission with high quantum yields of up to 0.6 in nonpolar environments. These dyes displayed a high brightness (up to 54,000 M(-1) cm(-1)). The promising structural and fluorescence properties of BASHY dyes fostered the preparation of non-cytotoxic, stable, and highly fluorescent poly(lactide-co-glycolide) nanoparticles that were effectively internalized by dendritic cells. The dyes were also shown to selectively stain lipid droplets in HeLa cells, without inducing any appreciable cytotoxicity or competing plasma membrane labeling; this confirmed their potential as fluorescent stains.

Cucurbiturils as supramolecular inhibitors of DNA restriction by type II endonucleases.

Cucurbiturils (CB6 and CB7) were shown to inhibit the enzymatically catalyzed restriction of plasmids and linear DNA. This effect can be inverted by supramolecular masking of the macrocycles through competitive complexation with polyamines. These experiments provide supramolecular control of biocatalytic processes.

A supramolecular keypad lock.

The reversible photoswitching between an anthracene derivative and its [4+4] dimer, using the template effect of the CB8 macrocycle, was demonstrated. This example of supramolecular chemistry in water was harnessed to demonstrate the operation of a keypad lock device that is driven by means of light and chemicals as inputs.

Aurora A triggers Lgl cortical release during symmetric division to control planar spindle orientation.

Mitotic spindle orientation is essential to control cell-fate specification and epithelial architecture. The tumor suppressor Lgl localizes to the basolateral cortex of epithelial cells, where it acts together with Dlg and Scrib to organize apicobasal polarity. Dlg and Scrib also control planar spindle orientation, but how the organization of polarity complexes is adjusted to control symmetric division is largely unknown. Here, we show that the Dlg complex is remodeled during Drosophila follicular epithelium cell division, when Lgl is released to the cytoplasm. Lgl redistribution during epithelial mitosis is reminiscent of asymmetric cell division, where it is proposed that Aurora A promotes aPKC activation to control the localization of Lgl and cell-fate determinants. We show that Aurora A controls Lgl localization directly, triggering its cortical release at early prophase in both epithelial and S2 cells. This relies on double phosphorylation within the putative aPKC phosphorylation site, which is required and sufficient for Lgl cortical release during mitosis and can be achieved by a combination of aPKC and Aurora A activities. Cortical retention of Lgl disrupts planar spindle orientation, but only when Lgl mutants that can bind Dlg are expressed. Hence, our work reveals that Lgl mitotic cortical release is not specifically linked to the asymmetric segregation of fate determinants, and we propose that Aurora A activation breaks the Dlg/Lgl interaction to allow planar spindle orientation during symmetric division via the Pins (LGN)/Dlg pathway.