How the COVID-19 Pandemic Affects the Provision of Psychotherapy: Results from Three Online Surveys on Austrian Psychotherapists.

This study aimed to assess patient numbers and the format in which psychotherapy was delivered by Austrian psychotherapists during different time points of the COVID-19 pandemic and to explore psychotherapists` experiences on pandemic-associated changes in their psychotherapeutic work as well as their wishes for support in their professional activities. Three cross-sectional online surveys were conducted between March 2020 and May 2022. The total number of participating psychotherapists was n = 1547 in 2020, n = 238 in 2021, and n = 510 in 2022. The number of patients treated was highest in 2022 and lowest at the beginning of the pandemic (p < 0.001). During the lockdown in 2020, only 25.0% of patients were treated in personal contact. This proportion increased in the following years, reaching 86.9% in 2022 (p < 0.001). After a substantial increase in the proportion of patients treated via the telephone and internet during the first lockdown, both proportions decreased during the pandemics' second and third year (p < 0.001). However, a larger proportion of patients were treated via the internet in 2022 compared to pre-pandemic times (p < 0.001). Psychotherapists reported that the pandemic affected mainly the setting in which psychotherapy was provided (29.6%), the working conditions and workload (27.1%), as well as the demand for psychotherapy (26.9%). About one-third of psychotherapists expressed support wishes for their psychotherapeutic work. Results suggest that the pandemic went along with a partial shift in the provision of psychotherapy towards psychotherapy via the internet but not the telephone. The increase in patient numbers and psychotherapists` reports of increased workload suggest a rise in the demand for mental health care during and in the aftermath of the pandemic.

On the formation of lipid droplets in human adipocytes: the organization of the perilipin-vimentin cortex.

We report on the heterogeneity and diversity of lipid droplets (LDs) in early stages of adipogenesis by elucidating the cell and molecular biology of amphiphilic and cytoskeletal proteins regulating and stabilizing the generation of LDs in human adipose cells. A plethora of distinct and differently sized LDs was detected by a brief application of adipocyte differentiation medium and additional short treatment with oleic acid. Using these cells and highly specific antibodies for LD-binding proteins of the perilipin (PLIN) family, we could distinguish between endogenously derived LDs (endogenous LDs) positive for perilipin from exogenously induced LDs (exogenous LDs) positive for adipophilin, TIP47 and S3-12. Having optimized these stimulation conditions, we used early adipogenic differentiation stages to investigate small-sized LDs and concentrated on LD-protein associations with the intermediate-sized filament (IF) vimentin. This IF protein was described earlier to surround lipid globules, showing spherical, cage-like structures. Consequently - by biochemical methods, by immunofluorescence microscopy and by electron- and immunoelectron microscopy - various stages of emerging lipid globules were revealed with perilipin as linking protein between LDs and vimentin. For this LD-PLIN-Vimentin connection, a model is now proposed, suggesting an interaction of proteins via opposed charged amino acid domains respectively. In addition, multiple sheaths of smooth endoplasmic reticulum cisternae surrounding concentrically nascent LDs are shown. Based on our comprehensive localization studies we present and discuss a novel pathway for the LD formation.

Lipid droplets, perilipins and cytokeratins--unravelled liaisons in epithelium-derived cells.

Lipid droplets (LDs) are spherical accumulations of apolar lipids and other hydrophobic substances and are generally surrounded by a thin cortical layer of specific amphiphilic proteins (APs). These APs segregate the LDs from the mostly polar components of the cytoplasm. We have studied LDs in epithelium-derived cell cultures and in particular characterized proteins from the perilipin (PLIN) gene family - in mammals consisting of the proteins Perilipin, Adipophilin, TIP47, S3-12 and MLDP/OXPAT (PLIN 1-5). Using a large number of newly generated and highly specific mono- and polyclonal antibodies specific for individual APs, and using improved LD isolation methods, we have enriched and characterized APs in greater detail and purity. The majority of lipid-AP complexes could be obtained in the top layer fractions of density gradient centrifugation separations of cultured cells, but APs could also be detected in other fractions within such separations. The differently sized LD complexes were analyzed using various biochemical methods and mass spectrometry as well as immunofluorescence and electron- in particular immunoelectron-microscopy. Moreover, by immunoprecipitation, protein-protein binding assays and by immunoelectron microscopy we identified a direct linkage between LD-binding proteins and the intermediate-sized filaments (IF) cytokeratins 8 and 18 (also designated as keratins K8 and K18). Specifically, in gradient fractions of higher density supposedly containing small LDs, we received as co-precipitations cytidylyl-, palmitoyl- and cholesterol transferases and other specific enzymes involved in lipid metabolism. So far, common proteomic studies have used LDs from top layer fractions only and did not report on these transferases and other enzymes. In addition to findings of short alternating hydrophobic/hydrophilic segments within the PLIN protein family, we propose and discuss a model for the interaction of LD-coating APs with IF proteins.

High post surgical opioid requirements in Crohn's disease are not due to a general change in pain sensitivity.

Crohn's disease (CD) is a painful inflammatory bowel disease with complex multigenic inheritance. Suggested on the basis of a few isolated reports CD patients require significantly higher post operative opioid doses than patients undergoing comparable severe abdominal surgery. Crohn's disease therefore may be a suitable model for the identification of novel pain susceptibility genes. In order to confirm this observation and to elucidate the underlying molecular mechanisms, we investigated if higher opioid needs of CD patients are due to a general change in pain sensitivity. Quantitative sensory testing (QST) was applied to a subgroup of patients and polymorphisms in the mu-opioid receptor (OPRM1) and catechol-O-methyltransferase (COMT) were investigated. Significantly increased post operative opioid requirements in CD patients were confirmed and QST assessment demonstrates that CD patients do not display increased pain sensitivity in terms of lowered thresholds to thermal and mechanical stimuli. The data also suggest that common variants in OPRM1 and specific 'high pain sensitivity'COMT haplotypes may not be the cause of high opioid needs. The results indicate that a more complex pathway is involved in the greater post operative opioid demand in CD. Therefore the presence of other, as yet unknown, genes could modulate opioid requirements in CD patients.

Novel actin-related proteins Arp-T1 and Arp-T2 as components of the cytoskeletal calyx of the mammalian sperm head.

The calyx is a large cytoskeletal component of the perinuclear theca of the mammalian sperm head, displaying remarkable morphological interspecies differences, which is biochemically characterized by resistance to high ionic strength and detergents and by a special protein composition, including the basic proteins calicin, cylicin I and II, and two major actin-capping proteins. In our calyx preparations from bull spermatozoa we have noted two major acidic components which upon partial amino acid sequencing have been identified as novel members of the subfamily of actin-related proteins (Arps). Antibodies raised against the corresponding human proteins, termed Arp-T1 and Arp-T2, have been used to detect the proteins by immunoblotting and immunofluorescence microscopy, demonstrating their specific synthesis in the testis, late in spermatid differentiation, and their localization in the calyx. The discovery of two novel Arps as major components in a cytoskeletal, nonmotile structure of mammalian spermatozoa suggests that certain members of this family of proteins may serve functions other than nucleation of actin filaments, and possible biological roles of such Arps in spermatozoa are discussed.