COVID-19 Pandemic: What We Have Learned on Stress, Technology Use, and Psychological Health Among University Students.

Nowadays, the medical COVID-19 emergency is ending worldwide, whereas there is an increasing need to understand the marks and the lessons learned from over two years of unprecedented changes in all life domains. Recent research has highlighted growing rates of psychological suffering among the general population as well as among university students, who were yet well-recognized as a vulnerable population even before the pandemic. The present work drives from university students' experience, and it aims to provide an overview of the key dimensions to take into account, in terms of main risks, changes, challenges, and resources, to develop updated tailored research and interventions promoting psychological health conditions in the current time within and beyond the educational context/university students' population. Implications for research and interventions are highlighted and discussed.

First isolation, identification, and pathogenicity evaluation of an EV-G6 strain in China.

Enterovirus G (EV-G) belongs to the Picornaviridae family and infects porcine populations worldwide. A total of 20 EV-G genotypes (EV-G1 to EV-G20) have been identified. In this study, we isolated and characterized an EV-G strain, named EV-G/YN29/2022, from the feces of diarrheic pigs. This was the first EV-G6 strain isolated in China. Comparison of the whole genome nucleotide and corresponding amino acid sequences showed that the isolate was more closely related to those of the EV-G6 genotype than other genotypes, with the complete genome sequence similarity ranging from 83.7% (Iba46442) to 84.4% (PEV-B-KOR), and corresponding amino acid homology ranged from 96% (Iba46442) to 96.8% (PEV-B-KOR). Similarly, the VP1 gene and corresponding amino acid sequences of EV-G/YN29/2022 were highly similar to those of the EV-G6 genotype (>82.9% and >94.3%, respectively). Thus, the isolated strain was classified as EV-G6 genotype. This was the first EV-G6 strain isolated in China. Pathogenicity analyses revealed that EV-G/YN29/2022 infection caused mild diarrhea, typical skin lesions, and weight reduction. The strain was mainly distributed to the intestinal tissue but was also found in the brain, mesenteric lymph nodes, spleen, and liver. Our results can be used as a reference to further elucidate the epidemiology, evolution, and pathogenicity of EV-G.

No more government-imposed societal-level COVID-19 control measures but still significant self-experienced burden for severely immunocompromised individuals - A cross-sectional survey in the Netherlands.

Objectives: In March 2023, all societal-level COVID-19 control measures were lifted by the Dutch government. This study was performed to understand the self-experienced burden of this new phase of COVID-19 on the perspectives and behaviors of severely immunocompromised individuals. Methods: This is an observational, descriptive, cross-sectional study in The Netherlands. An online survey was completed by severely immunocompromised individuals, to capture their general well-being (score from 1 = worst to 10 = best), mental and physical health, and daily and social activities during survey conduct and retrospectively for before onset of COVID-19. The survey was open for completion from May 24th until August 7nd, 2023. Results: Of the 236 respondents, 96.6 % had been vaccinated against COVID-19 and 24.6 % were shielding to avoid COVID-19 during survey conduct. The general well-being score for all respondents was 7.5 (±1.2 SD) before onset of the COVID-19 pandemic and 6.9 (±1.6 SD) during survey conduct (P<0.001). For the shielding group (n = 58), these scores were 7.6 (±1.0 SD) and 5.7 (±1.6 SD), respectively (P<0.001). Generally, for all questions about mental and physical health and daily and social activities, there was a trend towards more negative answers during survey conduct, compared with before onset of the COVID-19 pandemic, which was more pronounced for the shielding group. Conclusions: Despite absence of government-imposed societal measures, COVID-19 avoidance still had a self-experienced burden on perspectives and behaviors of immunocompromised individuals in The Netherlands, with a significantly lower general well-being during survey conduct, compared with before onset of COVID-19.

[Internet-based self-help for loneliness-a systematic review]. / Internetbasierte Selbsthilfe bei Einsamkeit ­ Eine systematische Übersicht.

BACKGROUND: Recently, the phenomenon of loneliness has received increasing attention. Loneliness is widespread and can have adverse consequences for mental and physical health if prolonged. Internet-based interventions (IBIs) for self-help have proven to be effective for a variety of psychological disorders. Due to several specific aspects, IBIs are also a relevant option for loneliness. This systematic review aims to present the current research on self-help IBIs for reducing loneliness. METHODS: A systematic literature search was conducted in the databases Web of Science, PubMed, Scopus, PsycInfo, MedLine, PsycIndex, Cochrane Library and PsyArXiv between December 2023 and early January 2024. We included original German or English studies that addressed IBIs for self-help to reduce loneliness. RESULTS: In total, eight studies published between 2017 and 2024 were included in the qualitative analysis. All studies were conducted in high-income countries, included predominantly well-educated female adults and were mostly satisfactory regarding their internal validity. DISCUSSION: The results of this review suggest that self-help IBIs may be a promising option for alleviating loneliness. However, the work also points to the need for further research. Future studies should consider larger samples and people of different ages, genders and education levels in order to generalise the results of the present review.

The effect of acute social isolation on neural molecular responses in components of the social decision-making network.

Prolonged or chronic social isolation has pronounced effects on animals, ranging from altered stress responses, increased anxiety and aggressive behaviour, and even increased mortality. The effects of shorter periods of isolation are much less well researched; however, short periods of isolation are used routinely for testing animal behaviour and physiology. Here, we studied how a 3 h period of isolation from a cagemate affected neural gene expression in three brain regions that contain important components of the social decision-making network, the hypothalamus, the nucleus taeniae of the amygdala, and the bed nucleus of the stria terminalis, using a gregarious bird as a model (zebra finches). We found evidence suggestive of altered neural activity, synaptic transmission, metabolism, and even potentially pain perception, all of which could create cofounding effects on experimental tests that involve isolating animals. We recommend that the effects of short-term social isolation need to be better understood and propose alternatives to isolating animals for testing.

Deep learning-based multimodal fusion of the surface ECG and clinical features in prediction of atrial fibrillation recurrence following catheter ablation.

BACKGROUND: Despite improvement in treatment strategies for atrial fibrillation (AF), a significant proportion of patients still experience recurrence after ablation. This study aims to propose a novel algorithm based on Transformer using surface electrocardiogram (ECG) signals and clinical features can predict AF recurrence. METHODS: Between October 2018 to December 2021, patients who underwent index radiofrequency ablation for AF with at least one standard 10-second surface ECG during sinus rhythm were enrolled. An end-to-end deep learning framework based on Transformer and a fusion module was used to predict AF recurrence using ECG and clinical features. Model performance was evaluated using areas under the receiver operating characteristic curve (AUROC), sensitivity, specificity, accuracy and F1-score. RESULTS: A total of 920 patients (median age 61 [IQR 14] years, 66.3% male) were included. After a median follow-up of 24 months, 253 patients (27.5%) experienced AF recurrence. A single deep learning enabled ECG signals identified AF recurrence with an AUROC of 0.769, sensitivity of 75.5%, specificity of 61.1%, F1 score of 55.6% and overall accuracy of 65.2%. Combining ECG signals and clinical features increased the AUROC to 0.899, sensitivity to 81.1%, specificity to 81.7%, F1 score to 71.7%, and overall accuracy to 81.5%. CONCLUSIONS: The Transformer algorithm demonstrated excellent performance in predicting AF recurrence. Integrating ECG and clinical features enhanced the models' performance and may help identify patients at low risk for AF recurrence after index ablation.

[Migration background and loneliness among middle-aged and older adults in Germany]. / Migrationshintergrund und Einsamkeit im mittleren und hohen Alter in Deutschland.

BACKGROUND: Currently, there is limited knowledge about the association between a migration background and loneliness among middle-aged and older individuals in Germany. The aim was therefore to examine the association between migration background and loneliness in this group. METHODS: Data were taken from the German Ageing Survey (Wave 7, November 2020 to March 2021), a representative sample of middle-aged and older individuals. The sample comprised 4145 individuals, and the mean age was 63.8 years. Of the respondents, 93.2% had no migration background, approximately 5.9% had a migration background with personal migration experience, and 0.9% had a migration background but no personal migration experience. The De Jong Gierveld tool was used to quantify loneliness. RESULTS: Multiple linear regressions showed that individuals with a migration background and their own migration experience have significantly higher levels of loneliness (ß = 0.15, 95% confidence interval (CI): 0.004 to 0.30, p < 0.05) compared to individuals without a migration background, whereas individuals with a migration background without their own migration experience have significantly lower levels of loneliness (ß = -0.27, 95% CI: -0.52 to -0.02 p < 0.05). CONCLUSIONS: Individuals with a migration background and their own migration experience appear to represent a risk group for high loneliness among middle-aged and older adults in Germany. In this respect, this group should be given special consideration in corresponding measures. Against the background of current (and potential future) migration movements, the results are of great importance as these groups in particular could be affected by loneliness.

Protocol to identify and isolate rare murine tumor-resident dendritic cell populations for low-input transcriptomic profiling.

Conventional type 1 dendritic cells (cDC1s) are critical for innate sensing of cancer, yet they are scarce in the tumor microenvironment (TME). Here, we present a protocol to identify and isolate cDC1 subsets from murine implantable tumors for subsequent transcriptomic profiling using a flow sorting-based strategy. We describe steps for cell culture of mouse tumors, tumoral growth, dissociation and isolation of tumoral cells, extracellular staining, and cell sorting. We then detail procedures for RNA isolation, mRNA library preparation, and sequencing. For complete details on the use and execution of this protocol, please refer to Papadas et al.1.

Loneliness and brain rhythmic activity in resting state: An exploratory report.

Recent studies using resting-state functional magnetic resonance imaging have shown that loneliness is associated with altered blood oxygenation in several brain regions. However, the relationship between loneliness and changes in neuronal rhythm activity in the brain remains unclear. To evaluate brain rhythm, we conducted an exploratory resting-state electroencephalogram (EEG) study of loneliness. We recorded resting-state EEG signals from 139 participants (94 women; mean age = 19.96 years) and analyzed power spectrum density (PSD) and functional connectivity (FC) in both the electrode and source spaces. The PSD analysis revealed significant correlations between loneliness scores and decreased beta-band powers, which may indicate negative emotion, attention, reward, and/or sensorimotor processing. The FC analysis revealed a trend of alpha-band FC associated with individuals' loneliness scores. These findings provide new insights into the neural basis of loneliness, which will facilitate the development of neurobiologically informed interventions for loneliness.

Cross-sectional association of food insecurity with loneliness in older adults: The role of sex, age, and psychosomatic factors.

OBJECTIVE: Food insecurity (FI) is a critical social determinant of poor psychosocial health. While data on the specific roles of sex and age in the FI-loneliness link among older adults are limited, the underlying mechanisms are largely unknown. This study examines the age-sex-specific associations of FI with loneliness among older adults in Ghana and quantifies the extent to which psychosomatic factors mediate the association. METHODS: We analyzed cross-sectional data from the Aging, Health, Psychological, and Health-seeking Behavior Study in Ghana. The past 30-day FI was assessed using items on hunger and breakfast skipping frequency due to a lack of resources. We assessed loneliness severity with the University of California, Los Angeles 3-item Loneliness Scale. Multivariable OLS regressions and bootstrapping mediation analysis using the Hayes PROCESS macro plug-in were used to evaluate the associations. RESULTS: We included 1,201 individuals aged ≥50 years (mean = 62.9 [SD = 11.9]; women = 63.3%). The prevalence of loneliness was 17.7%. The prevalence of moderate and severe FI was 44.0% and 8.5%, respectively. In the adjusted model, greater FI was significantly associated with loneliness severity (B = .22, SE = .029, p < .001). We found significant interactive effects of FI × age (B = -.17, SE = .023, p < .01) and FI × sex (B = -.28, SE = .036, p < .001) on loneliness. Thus, the FI-loneliness link was respectively more marked among women (B = .25, SE = .035, p < .001) and ≥65 age groups (B = .34, SE = .041, p < .001) than men (B = .16, SE = .051, p < .01) and those aged 50-64 (B = .22; SE = .040, p < .001). Finally, comorbid depression/anxiety (41.07%), hopelessness (48.6%), worthlessness (42.1%), functional limitations (8.2%), and pain severity (6.4%) mediated the FI-loneliness association. CONCLUSIONS: Age- and sex-specific associations between FI and loneliness exist among older Ghanaians. Addressing FI in concert with psychosomatic problems in older adults may contribute meaningfully to reducing loneliness in later life.

Optimization of pre-enrichment strategies for mouse hematopoietic stem cell isolation and metabolomic analysis.

Blood cell production arises from the activity of hematopoietic stem cells (HSCs), defined by their self-renewal capacity and ability to give rise to all mature blood cell types. The mouse remains one of the most studied species in hematological research, and markers to define and isolate mouse HSCs are well-established. Given the very low frequency of HSCs in the bone marrow, stem cell pre-enrichment by red blood cell lysis and magnetic cell separation is often performed as part of the isolation process to reduce sorting times. Several pre-enrichment strategies are available, differing in their speed, degree of enrichment, final cell yield and cost. In the current study, we performed a side-by-side comparison and provide a decision tree to help researchers select a pre-enrichment strategy for mouse HSC isolation depending on their downstream application. We then compared different pre-enrichment techniques in combination with metabolomics analysis of HSCs, where speed, yield and temperature during pre-enrichment are crucial factors, and found that the choice of pre-enrichment strategy significantly impacts the number of metabolites detected and levels of individual metabolites in HSCs.

Devising Isolation Forest-Based Method to Investigate the sRNAome of Mycobacterium tuberculosis Using sRNA-seq Data.

Small non-coding RNAs (sRNAs) regulate the synthesis of virulence factors and other pathogenic traits, which enables the bacteria to survive and proliferate after host infection. While high-throughput sequencing data have proved useful in identifying sRNAs from the intergenic regions (IGRs) of the genome, it remains a challenge to present a complete genome-wide map of the expression of the sRNAs. Moreover, existing methodologies necessitate multiple dependencies for executing their algorithm and also lack a targeted approach for the de novo sRNA identification. We developed an Isolation Forest algorithm-based method and the tool Prediction Of sRNAs using Isolation Forest for the de novo identification of sRNAs from available bacterial sRNA-seq data (http://posif.ibab.ac.in/). Using this framework, we predicted 1120 sRNAs and 46 small proteins in Mycobacterium tuberculosis. Besides, we highlight the context-dependent expression of novel sRNAs, their probable synthesis, and their potential relevance in stress response mechanisms manifested by M. tuberculosis.

Interventions for loneliness in older adults: a systematic review of reviews.

Loneliness in older persons is a major risk factor for adverse health outcomes. Before the COVID-19 pandemic led to unprecedented isolation and hampered programs aimed at preventing or reducing loneliness, many interventions were developed and evaluated. However, previous reviews provide limited or conflicting summaries of intervention effectiveness. This systematic review aimed to assess previous review quality and bias, as well as to summarize key findings into an overarching narrative on intervention efficacy. The authors searched nine electronic databases and indices to identify systematic reviews of interventions to reduce loneliness in older people prior to the COVID-19 pandemic; 6,925 records were found initially. Of these, 19 reviews met inclusion criteria; these encompassed 101 unique primary intervention studies that varied in research design, sample size, intervention setting, and measures of loneliness across 21 nations. While 42% of reviews had minimal risk of bias, only 8% of primary studies appraised similarly. Among the 101 unique articles reviewed, 63% of tested interventions were deemed by article author(s) as effective or partially effective. Generally, interventions that included animals, psychological therapies, and skill-building activities were more successful than interventions focused on social facilitation or health promotion. However, interventions that targeted multiple objectives aimed at reducing loneliness (e.g., improving social skills, enhancing social support, increasing social opportunities, and changing maladaptive social cognition) were more effective than single-objective interventions. Future programs should incorporate multiple approaches, and these interventions should be rigorously tested.

Protocol to isolate and characterize pulmonary-specific extracellular vesicles in mice.

Extracellular vesicles (EVs) are membranous nanoparticles classified based on their size and surface markers, which can be specific to various cell origins. Here, we present a protocol for the isolation of pulmonary-specific EVs in mice. We describe steps for differential centrifugation, density gradient centrifugation, and commercially available polyethylene glycol(PEG)-based precipitation, employing pulmonary-specific EV-bound chemicals and antibodies. We then detail procedures for the characterization of these EVs through nanoparticle tracking analysis, flow cytometry, scanning electron microscopy, and transmission electron microscopy. For complete details on the use and execution of this protocol, please refer to Lee et al.1,2,3,4.

A simplified laboratory approach for isolating M. oryzae spores from rice samples infected with multiple pathogens.

A method for separating M. oryzae from rice samples infected with multiple pathogens using basic laboratory equipment is described. We conducted a series of experiments to obtain a single spore of M. oryzae. This method can also be used to isolate spores from other fungal species.

Cancer Stem Cells: Detection and Characterization from Solid Tumors.

Cancer stem cells (CSCs) have emerged as an attractive research interest due to their prominent role in development of the tumors. CSCs are rare dormant cells that can self-renew and maintain tumor development and heterogeneity. A better understanding of CSCs can improve tumor classification and contribute toward the development of novel therapeutic approaches to fight cancer. Hence, it is of immense importance to comprehend the basic function of CSCs in tumor formation, which can only be possible by devising perfected methodologies to isolate, detect, and characterize them. In this chapter, we outline the key protocols to culture, identify, and isolate CSCs from solid tumors to further advance basic and clinical investigation related to CSCs and their role in tumor biology.

Generation of Bidimensional and Three-Dimensional Muscle Culture Systems.

Skeletal muscle is a postmitotic tissue composed of contractile myofibers that are oriented and connected to different layers of connective tissue. Nevertheless, adult muscle fibers retain the capacity to regenerate in response to damage, activating the classical muscle stem cell compartment, namely, satellite cells (SCs), which are mitotically quiescent cells until required for growth or repair and are localized between the basal lamina and sarcolemma of myofibers. The transition of SCs from the quiescent state toward activation, commitment, and differentiation involves the genetic and epigenetic adaptation to novel biological functions, entailing dynamic changes in the protein expression profile. Interestingly, some of the activities and signaling regulating proliferation, commitment, differentiation, and survival/apoptosis of satellite cells have been also partially recapitulated in vitro, taking advantage of robust markers, reliable techniques, and reproducible protocols. Over the years, different techniques of muscular cell culture have been designed including primary cultures from embryonic or postnatal muscle, myogenic cell line, and three-dimensional (3D) skeletal muscle construct. Typical two-dimensional (2D) muscle cell culture cannot fully recapitulate the complexity of living muscle tissues, restricting their usefulness for physiological studies. The development of functional 3D culture models represents a valid alternative to overcome the limitations of already available in vitro model, increasing our understanding of the roles played by the various cell types and how they interact. In this chapter, the development of bidimensional and three-dimensional cell cultures have been described, improving the technical aspect of satellite cell isolation, the best culture-based conditions for muscle cell growth and differentiation, and the procedures required to develop a three-dimensional skeletal muscle construct.

Development and characterization of an immortalized nasopharyngeal epithelial cell line to explore airway physiology and pathology in yak (Bos grunniens).

Airway epithelial cells play a crucial role in investigating the physiological and pathological mechanisms of the respiratory tract in yaks, a species whose unique respiratory system has garnered extensive interest. Despite this growing interest, there currently are no available airway epithelial cell lines from yaks, underscoring the crucial need to establish a yak respiratory epithelial cell line. Therefore, our objective was to isolate a population of primary yak nasopharyngeal epithelial cells (pYNE) and transform them into immortalized yak nasopharyngeal epithelial cells (iYNE), assessing their suitability as an in vitro model. Employing a combined method of physical elimination and differential adhesion, we successfully isolated a population of high-purity pYNE, and developed an iYNE line through pCI-neo-hTERT plasmid transfection. Karyotype and transmission electron microscopy analyses confirmed that pYNE and iYNE share identical morphologies and structures. Gel electrophoresis and real-time PCR analyses demonstrated that pYNE and iYNE expressed similar levels of KRT18 and CDH1 genes (p ≥ 0.541). Notably, iYNE expressed a significantly high level of TERT gene expression (p < 0.001). Immunofluorescence analysis demonstrated that both cell types expressed Pan-Cytokeratin, ZO-1, and E-cadherin proteins. Furthermore, immunoblotting analysis indicated significantly higher levels of hTERT and Ki67 proteins in iYNE (p < 0.001), and similar levels of Cluadin-3 and Occludin proteins (p ≥ 0.103). Proliferation curve analysis highlighted iYNE's serum-dependency and significantly enhanced proliferation capacities (p < 0.001). Additionally, pYNE and iYNE cells demonstrated comparable susceptibilities to infectious bovine rhinotracheitis virus (IBRV). These findings collectively suggest that the developed iYNE retains the evaluated physiological characteristics of pYNE, making it an appropriate in vitro model. This advancement will facilitate further investigation into the respiratory physiological and pathological mechanisms in yaks.

Recovery from social isolation requires dopamine in males, but not the autism-related gene nlg3 in either sex.

Social isolation causes profound changes in social behaviour in a variety of species. However, the genetic and molecular mechanisms modulating behavioural responses to social isolation and social recovery remain to be elucidated. Here, we quantified the behavioural response of vinegar flies to social isolation using two distinct protocols (social space preference and sociability, the spontaneous tendencies to form groups). We found that social isolation increased social space and reduced sociability. These effects of social isolation were reversible and could be reduced after 3 days of group housing. Flies with a loss of function of neuroligin3 (orthologue of autism-related neuroligin genes) with known increased social space in a socially enriched environment were still able to recover from social isolation. We also show that dopamine (DA) is needed for a response to social isolation and recovery in males but not in females. Furthermore, only in males, DA levels are reduced after isolation and are not recovered after group housing. Finally, in socially enriched flies mutant for neuroligin3, DA levels are reduced in males, but not in females. We propose a model to explain how DA and neuroligin3 are involved in the behavioural response to social isolation and its recovery in a dynamic and sex-specific manner.