Psychiatric Hospital of Leros: a portrayal of the current situation.

The purpose of this study was to describe the demographic and clinical features of the inpatients currently residing at the Psychiatric Hospital of Leros. The present systematic documentation and presentation aimed to demonstrate the standard of living and healthcare conditions provided today, after the implementation of the State's "Psychargos" program; this is the main Greek Psychiatric reform program, adhering to the principles of deinstitutionalization and community psychiatry, in accordance with the current international guidelines. In addition, we discussed the current relationship between the psychiatric departments of the hospital and the other departments and clinics in terms of providing healthcare services to chronic psychiatric inpatients in full compliance with the biopsychosocial model and its application to the unique case of Leros. The implemented patient profiles incorporated both subjective and objective factors, such as compliance with rules and treatment, self-injury, and harm to others. Furthermore, we quantified and categorized the level of care required for each patient in terms of personnel-reported activities. This parameter was assessed through the Greek version of Katz's Index of Independence in Activities of Daily Living. Simultaneously, the fundamental actions provided to inpatients by the social care and support services of the hospital were also depicted and categorized, in terms of connection to State social services, communication with the patient's families, and cooperation between the families and the hospital for the patient's healthcare needs. Furthermore, we analyzed and presented all statistically significant correlations found in our patients' characteristics. Briefly, the main results of our study show that the mean age of the 212 patients was 62.4 years old (with a standard deviation of ±13.6 years and the longest hospitalization of 62 consecutive years) including patients from the institution's asylum period. Since 1989, the year when the psychiatric reform began in our hospital, 87 new patients were admitted, 85.1% of whom were from the southern Aegean, thus following the principle of naiveness. Intellectual disabilities and psychotic spectrum disorders were the most common disorders among the total number of hospitalized patients, accounting for 40% in each category. Regarding the 87 patients hospitalized after 1989, psychotic spectrum disorders were diagnosed in the vast majority (58 patients, 66.7%) followed by organic mental disorders (10 patients, 11.5%). The rest were diagnosed with other disorders. Somatic comorbidity and the need for care and services, especially for patients with intellectual disabilities, demonstrate how the Institution now mainly offers psychogeriatric healthcare services. In conclusion, the purpose of this study was to highlight the Psychiatric Hospital of Leros as it stands today, in stark contrast to the long-established, stereotypical depiction of asylums in the scientific and public communities.

The interactome of the UapA transporter reveals putative new players in anterograde membrane cargo trafficking.

Neosynthesized plasma membrane (PM) proteins co-translationally translocate to the ER, concentrate at regions called ER-exit sites (ERes) and pack into COPII secretory vesicles which are sorted to the early-Golgi through membrane fusion. Following Golgi maturation, membrane cargoes reach the late-Golgi, from where they exit in clathrin-coated vesicles destined to the PM, directly or through endosomes. Post-Golgi membrane cargo trafficking also involves the cytoskeleton and the exocyst. The Golgi-dependent secretory pathway is thought to be responsible for the trafficking of all major membrane proteins. However, our recent findings in Aspergillus nidulans showed that several plasma membrane cargoes, such as transporters and receptors, follow a sorting route that seems to bypass Golgi functioning. To gain insight on membrane trafficking and specifically Golgi-bypass, here we used proximity dependent biotinylation (PDB) coupled with data-independent acquisition mass spectrometry (DIA-MS) for identifying transient interactors of the UapA transporter. Our assays, which included proteomes of wild-type and mutant strains affecting ER-exit or endocytosis, identified both expected and novel interactions that might be physiologically relevant to UapA trafficking. Among those, we validated, using reverse genetics and fluorescence microscopy, that COPI coatomer is essential for ER-exit and anterograde trafficking of UapA and other membrane cargoes. We also showed that ArfAArf1 GTPase activating protein (GAP) Glo3 contributes to UapA trafficking at increased temperature. This is the first report addressing the identification of transient interactions during membrane cargo biogenesis using PDB and proteomics coupled with fungal genetics. Our work provides a basis for dissecting dynamic membrane cargo trafficking via PDB assays.

Phytochemical investigation of Pistacia lentiscus L. var. Chia leaves: A byproduct with antimicrobial potential.

Pistacia lentiscus L. var. Chia belongs to the Anacardiaceae family, and it is cultivated only in the south part of Chios island, in Greece. Even though it is renowned for its unique resin, Chios mastic gum (CMG), the tree leaves have also been used in traditional medicine, while the annual pruning generates a large biomass of unused by-products. Thus, the aim of the present study was the detailed phytochemical investigation of P. lentiscus var. Chia leaves towards the search of antimicrobial agents. UPLC-HRMS & HRMS/MS based dereplication methods led to the detailed characterization of the aqueous leaf extract. In addition, twelve compounds were isolated and purified from the methanol extract and were identified using spectroscopic and spectrometric methods (NMR, HRMS) belonging to phenolic acids, tannins, flavonoids and terpenes, with the most interesting being 2-hydroxy-1,8-cineole ß-D-glucopyranoside which was isolated for the first time in the Anacardiaceae family. Remarkably, based on NMR data, methanol and aqueous extracts were found to be particularly rich in shikimic acid, a valuable building block for the pharmaceutical industry, for instance in the synthesis of the active ingredient of Tamiflu®, oseltamivir. Finally, extracts (EtOAc, MeOH, H2O) and major compounds i.e., shikimic acid, 2-hydroxy-1,8-cineole ß-D-glucopyranoside and myricitrin were evaluated for their antimicrobial properties. MeOH and H2O mastic leaf extracts as well as myricitrin and, particularly, 2-hydroxy-1,8-cineole ß-D-glucopyranoside showed significant selective activity against pathogenic Mucorales, but not against Aspergilli (Aspergillus nidulans, Aspergillus fumigatus), Candida albicans or bacteria (Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis).

Golgi-Bypass Is a Major Unconventional Route for Translocation to the Plasma Membrane of Non-Apical Membrane Cargoes in Aspergillus nidulans.

Nutrient transporters have been shown to translocate to the plasma membrane (PM) of the filamentous fungus Aspergillus nidulans via an unconventional trafficking route that bypasses the Golgi. This finding strongly suggests the existence of distinct COPII vesicle subpopulations, one following Golgi-dependent conventional secretion and the other directed towards the PM. Here, we address whether Golgi-bypass concerns cargoes other than nutrient transporters and whether Golgi-bypass is related to cargo structure, size, abundance, physiological function, or polar vs. non-polar distribution in the PM. To address these questions, we followed the dynamic subcellular localization of two selected membrane cargoes differing in several of the aforementioned aspects. These are the proton-pump ATPase PmaA and the PalI pH signaling component. Our results show that neosynthesized PmaA and PalI are translocated to the PM via Golgi-bypass, similar to nutrient transporters. In addition, we showed that the COPII-dependent exit of PmaA from the ER requires the alternative COPII coat subunit LstA, rather than Sec24, whereas PalI requires the ER cargo adaptor Erv14. These findings strengthen the evidence of distinct cargo-specific COPII subpopulations and extend the concept of Golgi-independent biogenesis to essential transmembrane proteins, other than nutrient transporters. Overall, our findings point to the idea that Golgi-bypass might not constitute a fungal-specific peculiarity, but rather a novel major and cargo-specific sorting route in eukaryotic cells that has been largely ignored.

On the Evidence Supporting That AN11127 Encodes an Aspergillus Nidulans Sec12 Orthologous Protein. Reply to Bravo-Plaza et al. Comment on "Dimou et al. Profile of Membrane Cargo Trafficking Proteins and Transporters Expressed under N Source Derepressing Conditions in Aspergillus nidulans. J. Fungi 2021, 7, 560".

Prof. Peñalva and co-workers provided evidence that AN11127 is related by sequence and function to Sec12 [...].

Profile of Membrane Cargo Trafficking Proteins and Transporters Expressed under N Source Derepressing Conditions in Aspergillus nidulans.

Solute and ion transporters are proteins essential for cell nutrition, detoxification, signaling, homeostasis and drug resistance. Being polytopic transmembrane proteins, they are co-translationally inserted and folded into the endoplasmic reticulum (ER) of eukaryotic cells and subsequently sorted to their final membrane destination via vesicular secretion. During their trafficking and in response to physiological/stress signals or prolonged activity, transporters undergo multiple quality control processes and regulated turnover. Consequently, transporters interact dynamically and transiently with multiple proteins. To further dissect the trafficking and turnover mechanisms underlying transporter subcellular biology, we herein describe a novel mass spectrometry-based proteomic protocol adapted to conditions allowing for maximal identification of proteins related to N source uptake in A. nidulans. Our analysis led to identification of 5690 proteins, which to our knowledge constitutes the largest protein dataset identified by omics-based approaches in Aspergilli. Importantly, we detected possibly all major proteins involved in basic cellular functions, giving particular emphasis to factors essential for membrane cargo trafficking and turnover. Our protocol is easily reproducible and highly efficient for unearthing the full A. nidulans proteome. The protein list delivered herein will form the basis for downstream systematic approaches and identification of protein-protein interactions in living fungal cells.

Life and Death of Fungal Transporters under the Challenge of Polarity.

Eukaryotic plasma membrane (PM) transporters face critical challenges that are not widely present in prokaryotes. The two most important issues are proper subcellular traffic and targeting to the PM, and regulated endocytosis in response to physiological, developmental, or stress signals. Sorting of transporters from their site of synthesis, the endoplasmic reticulum (ER), to the PM has been long thought, but not formally shown, to occur via the conventional Golgi-dependent vesicular secretory pathway. Endocytosis of specific eukaryotic transporters has been studied more systematically and shown to involve ubiquitination, internalization, and sorting to early endosomes, followed by turnover in the multivesicular bodies (MVB)/lysosomes/vacuole system. In specific cases, internalized transporters have been shown to recycle back to the PM. However, the mechanisms of transporter forward trafficking and turnover have been overturned recently through systematic work in the model fungus Aspergillus nidulans. In this review, we present evidence that shows that transporter traffic to the PM takes place through Golgi bypass and transporter endocytosis operates via a mechanism that is distinct from that of recycling membrane cargoes essential for fungal growth. We discuss these findings in relation to adaptation to challenges imposed by cell polarity in fungi as well as in other eukaryotes and provide a rationale of why transporters and possibly other housekeeping membrane proteins 'avoid' routes of polar trafficking.

Translocation of nutrient transporters to cell membrane via Golgi bypass in Aspergillus nidulans.

Nutrient transporters, being polytopic membrane proteins, are believed, but not formally shown, to traffic from their site of synthesis, the ER, to the plasma membrane through Golgi-dependent vesicular trafficking. Here, we develop a novel genetic system to investigate the trafficking of a neosynthesized model transporter, the well-studied UapA purine transporter of Aspergillus nidulans. We show that sorting of neosynthesized UapA to the plasma membrane (PM) bypasses the Golgi and does not necessitate key Rab GTPases, AP adaptors, microtubules or endosomes. UapA PM localization is found to be dependent on functional COPII vesicles, actin polymerization, clathrin heavy chain and the PM t-SNARE SsoA. Actin polymerization proved to primarily affect COPII vesicle formation, whereas the essential role of ClaH seems indirect and less clear. We provide evidence that other evolutionary and functionally distinct transporters of A. nidulans also follow the herein identified Golgi-independent trafficking route of UapA. Importantly, our findings suggest that specific membrane cargoes drive the formation of distinct COPII subpopulations that bypass the Golgi to be sorted non-polarly to the PM, and thus serving house-keeping cell functions.

The peroxisomal SspA protein is redundant for purine utilization but essential for peroxisome localization in septal pores in Aspergillus nidulans.

In an in silico search for correlated gene loss with fungal peroxisomal uric acid oxidase (UOX), we identified PMP22-like proteins, some of which function as promiscuous channels in organellar membranes. To investigate whether PMP22 channels have a role in peroxisomal uric acid transport and catabolism, we functionally analyzed the closest homologue in Aspergillus nidulans, named SspA. We confirmed that SspA is a peroxisomal membrane protein that co-localizes significantly with PTS1-tagged mRFP, UOX or HexA, the latter considered a protein of Woronin bodies (WB), organelles originating from peroxisomes that dynamically plug septal pores in ascomycetes. Our results suggest that in A. nidulans, unlike some other ascomycetes, there is no strict protein segregation of peroxisomal and WB-specific proteins. Importantly, genetic deletion of sspA, but not of hexA, led to lack of peroxisomal localization at septal pores, suggesting that SspA is a key factor for septal pore functioning. Additionally, ΔsspA resulted in increased sensitivity to oxidative stress, apparently as a consequence of not only the inability to plug septal pores, but also a recorded reduction in peroxisome biogenesis. However, deleting sspA had no effect on uric acid or purine utilization, as we hypothesized, a result also in line with the observation that expression of SspA was not affected by regulatory mutants and conditions known to control purine catabolic enzymes. Our results are discussed within the framework of previous studies of SspA homologues in other fungi, as well as, the observed gene losses of PMP22 and peroxisomal uric acid oxidase.

Evaluation of tubulointerstitial lesions' severity in patients with glomerulonephritides: an NMR-based metabonomic study.

An 1H NMR-based metabonomic approach was used to investigate the correlation of histopathologically assessed tubulointerstitial lesions with the urinary metabolite profile in 77 patients with glomerulonephritides submitted to renal biopsy. The presence of renal damage was predicted with a sensitivity of 96% and a specificity of 99%. Patients with mild, moderate, and severe tubulointerstitial lesions were progressively differentiated from the healthy individuals in the Orthogonal Signal Correction Partial Least-Squares-Discriminant Analysis (OSC/PLS-DA) models with a statistically significant separation between those with mild and with severe lesions. The onset of the tubulointerstitial lesions is characterized by decreased excretion of citrate, hippurate, glycine, and creatinine, whereas further deterioration is followed by glycosuria, selective aminoaciduria, total depletion of citrate and hippurate, and gradual increase in the excretion of lactate, acetate, and trimethylamine-N-oxide. NMR-based metabonomic urinalysis could contribute to the early evaluation of the severity of the renal damage and possibly to the monitoring of kidney function.

Zosteriform cutaneous metastases from breast adenocarcinoma.

Cutaneous metastases from breast adenocarcinoma are usually nodular, single or multiple. Their zosteriform distribution is very rare. We present a 54-year-old woman with cutaneous zosteriform nodular metastases on the right side of her thorax, and infiltration of the corresponding arm, 3 months after the excision of adenocarcinoma of her right breast.