Unique chromoplast organisation and carotenoid gene expression in carotenoid-rich carrot callus

Main conclusion The new model orange callus line, similar to carrot root, was rich in carotenoids due to altered expression of some carotenogenesis-associated genes and possessed unique diversity of chromoplast ultrastructure. Callus induced from carrot root segments cultured in vitro is usually pale yellow (p-y) and poor in carotenoids. A unique, non-engineered callus line of dark orange (d-o) colour was developed in this work. The content of carotenoid pigments in d-o callus was at the same level as in an orange carrot storage root and nine-fold higher than in p-y callus. Carotenoids accumulated mainly in abundant crystalline chromoplasts that are also common in carrot root but not in p-y callus. Using transmission electron microscopy, other types of chromoplasts were also found in d-o callus, including membranous chromoplasts rarely identified in plants and not observed in carrot root until now. At the transcriptional level, most carotenogenesis-associated genes were upregulated in d-o callus in comparison to p-y callus, but their expression was downregulated or unchanged when compared to root tissue. Two pathway steps were critical and could explain the massive carotenoid accumulation in this tissue. The geranylgeranyl diphosphate synthase gene involved in the biosynthesis of carotenoid precursors was highly expressed, while the β-carotene hydroxylase gene involved in β-carotene conversion to downstream xanthophylls was highly repressed. Additionally, paralogues of these genes and phytoene synthase were differentially expressed, indicating their tissue-specific roles in carotenoid biosynthesis and metabolism. The established system may serve as a novel model for elucidating plastid biogenesis that coincides with carotenogenesis

Inhibition of carotenoid biosynthesis by CRISPR/Cas9triggers cell wall remodelling in carrot

Recent data indicate that modifications to carotenoid biosynthesis pathway in plants alter the expression of genes affecting chemical composition of the cell wall. Phytoene synthase (PSY) is a rate limiting factor of carotenoid biosynthesis and it may exhibit species-specific and organ-specific roles determined by the presence of psy paralogous genes, the importance of which often remains unrevealed. Thus, the aim of this work was to elaborate the roles of two psy paralogs in a model system and to reveal biochemical changes in the cell wall of psy knockout mutants. For this purpose, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR associated (Cas9) proteins (CRISPR/Cas9) vectors were introduced to carotenoid-rich carrot (Daucus carota) callus cells in order to induce mutations in the psy1 and psy2 genes. Gene sequencing, expression analysis, and carotenoid content analysis revealed that the psy2 gene is critical for carotenoid biosynthesis in this model and its knockout blocks carotenogenesis. The psy2 knockout also decreased the expression of the psy1 paralog. Immunohistochemical staining of the psy2 mutant cells showed altered composition of arabinogalactan proteins, pectins, and extensins in the mutant cell walls. In particular, low-methylesterified pectins were abundantly present in the cell walls of carotenoidrich callus in contrast to the carotenoid-free psy2 mutant. Transmission electron microscopy revealed altered plastid transition to amyloplasts instead of chromoplasts. The results demonstrate for the first time that the inhibited biosynthesis of carotenoids triggers the cell wall remodelling

sEMG and vibration system monitoring for differential diagnosis in temporomandibular joint disorders

The stomatognathic system represents an important element of human physiology, constituting a part of the digestive, respiratory, and sensory systems. One of the signs of temporomandibular joint disorders (TMD) can be the formation of vibroacoustic and electromyographic (sEMG) phenomena. The aim of the study was to evaluate the effectiveness of temporomandibular joint rehabilitation in patients suffering from locking of the temporomandibular joint (TMJ) articular disc by analysis of vibrations, sEMG registration of masseter muscles, and hypertension of masticatory muscles. In this paper, a new system for the diagnosis of TMD during rehabilitation is proposed, based on the use of vibration and sEMG signals. The operation of the system was illustrated in a case study, a 27-year-old woman with articular dysfunction of the TMJ. The first results of TMD diagnostics using the k-nearest neighbors method are also presented on a group of fifteen people (ten women and five men). Vibroacoustic registration of temporomandibular joints, sEMG registration of masseter muscles, and functional manual analysis of the TMJ were simultaneously assessed before employing splint therapy with stomatognathic physiotherapy. Analysis of vibrations with the monitoring of sEMG in dysfunctions of the TMJ can lead to improve differential diagnosis and can be an objective way of monitoring the rehabilitation process of TMD

Analiza zmian w zakresie zaspokojenia i frustracji podstawowych potrzeb psychologicznych oraz stresu i radzenia sobie z nim w pierwszych miesiącach pandemii COVID-19 w 2020 roku wśród kobiet

Globalna pandemia COVID-19 była wydarzeniem powodującym liczne deprywacje, zagrożenia oraz konieczność wprowadzenia drastycznych restrykcji. Celem badania było sprawdzenie, czy zmieniało się zaspokojenie i frustracja podstawowych potrzeb psychologicznych, odczuwanie stresu i radzenie sobie z nim wraz z przebiegiem pandemii i zmianami sytuacji społeczno-ekonomicznej. Badanie przeprowadzono na kobietach, które – według dostępnych danych – doświadczały większego stresu i frustracji związanej z sytuacją pandemii. W okresie od marca do maja 2020 roku dwukrotnie w odstępie dwóch miesięcy zbadano on-line 298 ochotniczek (wiek 16–73) za pomocą BPNS&FS Chen i in. (2015), PSS Cohena i in. (1983), COPE Carvera i in. (1989) oraz zestawem pytań związanych specyficznie z pandemią. Stwierdzono wzrost frustracji potrzeb kompetencji i więzi. Obniżył się poziom odczuwanego stresu i wzrosło poczucie bezpieczeństwa. Nasiliły się strategie tłumienia konkurencyjnych działań, pozytywnej reinterpretacji i rozwoju oraz zaprzeczania, natomiast osłabiła się strategia psychicznego uwalnianie się. Zmianom uległo także wiele specyficznych dla sytuacji pandemii strategii radzenia sobie. Pomimo utrzymywania się wysokiego stresu specyficznego, generalnie nastąpił spadek działań prewencyjnych i przestrzegania zasad ochrony przed zakażeniem. Wyniki wskazują na deprymujący i stresujący wpływ sytuacji pandemii na kobiety oraz na dynamiczne zmiany adaptacyjne i radzenia sobie z tą sytuacją

Basic Psychological Needs, Stress and Coping Strategies - A Comparative Analysis of Pre-Epidemic and Epidemic Experiences

Background: The global epidemic of SARS-CoV-2 caused various deprivations and threats and forced the implementation of drastic restrictions in the whole world, including Poland. Objectives: Recognizing the consequences of the epidemic in the context of basic psychological needs satisfaction and frustration, the level of experienced stress, and the use of coping strategies. Design: The results of the pre-epidemic group (N = 626; aged 18 - 40) were compared with the results of the epidemic group (N = 282; aged 17 - 44). The following tests were used; BPNS&FS (Chen et al., 2015), PSS (Cohen et al., 1983), and COPE (Carver et al., 1989). Results: Women from the epidemic group reported higher levels of stress, lower satisfaction and higher frustration of autonomy and competence than the control group. Men from the epidemic group differed from the control group only by a higher frustration of autonomy. We found significant differences in coping strategy preference. The epidemic group was characterized by the following strategies: acceptance, mental disengagement, restraint, positive reinterpretation and growth, use of emotional social support, and use of humor. Conclusions: The results are consistent with previously documented reactions to uncontrolled and critical stressors and indicate the adaptability of undertaken coping efforts