A subcellular proteome atlas of the yeast Komagataella phaffii.

The compartmentalization of metabolic and regulatory pathways is a common pattern of living organisms. Eukaryotic cells are subdivided into several organelles enclosed by lipid membranes. Organelle proteomes define their functions. Yeasts, as simple eukaryotic single cell organisms, are valuable models for higher eukaryotes and frequently used for biotechnological applications. While the subcellular distribution of proteins is well studied in Saccharomyces cerevisiae, this is not the case for other yeasts like Komagataella phaffii (syn. Pichia pastoris). Different to most well-studied yeasts, K. phaffii can grow on methanol, which provides specific features for production of heterologous proteins and as a model for peroxisome biology. We isolated microsomes, very early Golgi, early Golgi, plasma membrane, vacuole, cytosol, peroxisomes and mitochondria of K. phaffii from glucose- and methanol-grown cultures, quantified their proteomes by liquid chromatography-electrospray ionization-mass spectrometry of either unlabeled or tandem mass tag-labeled samples. Classification of the proteins by their relative enrichment, allowed the separation of enriched proteins from potential contaminants in all cellular compartments except the peroxisomes. We discuss differences to S. cerevisiae, outline organelle specific findings and the major metabolic pathways and provide an interactive map of the subcellular localization of proteins in K. phaffii.

Benign uterine mass-discrimination from leiomyosarcoma by a preoperative risk score: a multicenter cohort study.

PURPOSE: Discrimination of uterine leiomyosarcoma (LMS) and leiomyoma (LM) prior to surgery by basic preoperative characteristics and development of a preoperative leiomyosarcoma score. METHODS: A predominantly prospective cohort of 826 patients with LM from a clinical institution and an outpatient center was included in the study. Further a predominantly retrospective cohort of 293 patients with LMS was included from the counseling database of the German Clinical Center of Excellence for Genital Sarcoma and Mixed Tumors (DKSM, University Medicine Greifswald, Germany). We analyzed and compared anamnestic, epidemiological and clinical findings between both cohorts. Tenfold cross-validated logistic regression and random forest was performed on the 80% training set. The preoperative LMS score (pLMS) was developed based on logistic regression and independently evaluated by analyzing the area under the receiver operating characteristic curve (AUC) with the 20% test set. RESULTS: In the LMS cohort, 63.1% had initially surgery for presumed LM and only 39.6% of endometrial biopsies revealed LMS. Key features for LMS discrimination were found to be bleeding symptoms: intermenstrual bleeding [RRc = 2.71, CI = (1.90-3.49), p < 0.001], hypermenorrhea [RRc = 0.28, CI = (0.15-0.50), p < 0.001], dysmenorrhea [RRc = 0.22, CI = (0.10-0.51), p < 0.001], postmenstrual bleeding [RRc = 2.08, CI = (1.30-2.75), p < 0.001], suspicious sonography [RRc = 1.21, CI = (1.19-1.22), p < 0.001] and the tumor diameter (each centimeter difference: ß = 0.24, SD = 0.04, p < 0.001). pLMS achieved a mean cross-validated AUC of 0.969 (SD = 0.019) in the training set and an AUC of 0.968 in the test set. CONCLUSIONS: The presented score is based on basic clinical characteristics and allows the prediction of LMS prior to a planned surgery of a uterine mass. In case pLMS is between - 3 and + 1, we suggest subsequent diagnostics, such as endometrial biopsy, color Doppler sonography, LDH measurement, MRI and transcervical biopsy.

Skin toxicities and unmet supportive care needs of patients with cancer undergoing EGFR-inhibitor therapy / Toxische Hautreaktionen und Unterstützungsbedarf von KrebspatientInnen im Zuge einer EGFR-Antikörpertherapie. Eine deskriptive Studie

Background: Targeted therapies, such as the EGFR (epidermal growth factor receptor) inhibitor therapy, are being used to treat patients with various solid and metastatic tumours. Skin toxicities are a common side effect of this therapy. Aim: The aim of this study was to assess the effects of skin toxicities on quality of life of patients with cancer undergoing EGFR inhibitor therapy, as well as their unmet supportive care needs. Method: Embedded design. A standardised quantitative survey was administered and analysed. In addition, memos and audiotaped material of insightful conversations with the patients after survey administration were included in the analyses. Results: Among the three domains of the effects of skin toxicities on quality of life, physical symptoms (e. g. itching skin, rash) were most frequently reported to impair quality of life, while associated emotional and functional impairments were less frequently reported. Patients don't consider the management of skin toxicities to be a priority during their treatment, skin toxicities are rather perceived in context of the total symptom burden. Yet, we observed significant correlations between the assessed quality of life and unmet supportive care need domains, especially concerning physical and psychological needs. Conclusions: Although no clinically significant impairment of quality of life of patients undergoing EGFR inhibitor therapy was found, skin changes should be addressed in supportive interventions embedded in routine symptom management.

Prevalence and effects of caregiving on children.

AIM: This paper reports the prevalence and its related sociodemographic factors of informal caregiving by underage children in Austria. The quantity and intensity of caregiving activities, the motivation for and effects of caregiving and how this differs from non-caregiving children were investigated. BACKGROUND: Young carers are a worldwide phenomenon. Due to methodological and sampling problems, little quantitative data are available. DESIGN: Cross-sectional, descriptive study. METHODS: Based on a random selection of 85 schools and 474 classes, a total of 7403 children aged 10-14 years completed a self-reporting questionnaire that asked for children's help in their families. Descriptive and inferential statistics were used to analyse the data. RESULTS: In the sample, 4·5% caregiving children were identified. The average age of young carers was 12·5 years. Most young carers were female (69·8% vs. 52·7% in the non-young carers group). Young carers assumed more responsibilities (household tasks, general care and sibling care) than their peers. They showed a higher level of physical (e.g. headache 38·2% vs. 24·4%) and mental (e.g. to worry about 68·1% vs. 41·8%) adverse effects than non-young carers. Extrapolation suggests a rate of 3·5% young carers in underage children of 5-18 years in Austria. CONCLUSION: Data on national level are essential preconditions to initiate support for young carers. Nurses can promote children's health and well-being through prevention of an inappropriate caregiving role.

Carbon isotope ratio determination and investigation of seized testosterone preparations.

In the present study, the content of a number of black market testosterone products collected in Austria has been analyzed. Additionally, (13) C/(12) C ratios were measured for testosterone in the products after cleavage of the testosterone ester. The aim was to determine whether some of these products had similar (13) C/(12) C ratios to those normally found for endogenous testosterone, which could prevent a positive isotopic ratio mass spectrometric (IRMS) finding in doping control. Moreover, it was investigated to what extent the preparations contained the masking agent epitestosterone, in order to lower the testosterone/epitestosterone (T/E) ratio in urinary steroid profiles. Out of 30 analyzed products, the declared ingredients differed from the actual content in 10 cases. Epitestosterone, however, could not be found in any of the products. The products displayed δ(13)C(VPDB) values between -23.6 and -29.4‰. For more than half of these products, the values were within a range reported for endogenous urinary steroids.

Etiology and site of temporal lobe epilepsy influence postictal cytokine release.

Inflammatory mechanisms are involved in the pathogenesis of epilepsy. Vice versa, immune functions are regulated by the brain. We measured postictal changes in serum levels of the immuno-modulating cytokines IL-1beta, IL-6 and TNFalpha in patients with well-defined temporal lobe epilepsy (TLE) and determined modifying factors. Serum levels of IL-1beta, IL-6 and TNFalpha were quantified by ELISA at baseline as well as immediately, 1h and 24h after a complex partial (CPS) or secondary generalized tonic-clonic seizure (GTCS) during video-EEG monitoring in 25 patients suffering from temporal epilepsy. IL-6 increased by 51% immediately after the seizure (p<0.01) and remained elevated for 24h. This increase lacked in patients with hippocampal sclerosis (HS; n=16, mean increase 28%, p>0.5, vs. 112%, p<0.01 in patients without HS). IL-6 levels were higher after right-sided seizures as compared to left-sided seizures 24h after the seizure (8.7pg/mL vs. 3.4pg/mL, p<0.05). In patients taking valproate (VPA, n=9), the levels of IL-1beta were higher as compared to patients not treated with VPA. The results suggest a relationship between the cytokine system and characteristics of TLE such as side and pathology.

NK and CD4+ T cell changes in blood after seizures in temporal lobe epilepsy.

UNLABELLED: Immunological phenomena may affect the course of focal epilepsy. We analyzed prospectively the pre- and postictal distribution of leukocyte subsets in epileptic patients. METHODS: Twenty-two patients (age 36.6+/-10.8 years, 50% men) with temporal lobe epilepsy were included. Distribution of leukocyte subsets and serum levels of epinephrine were measured in peripheral blood immediately and 24 h after seizures and compared to baseline values. RESULTS: In the immediate postictal state (10+/-6 min), we observed a significant relative increase of total leukocytes (42%, p=0.0004), neutrophil leukocytes (55%, p=0.0007), total lymphocytes (45%, p=0.0019), natural killer (NK) cells (104%, p=0.0017), and epinephrine (454%, p=0.0014). CD4(+) T cells decreased by 13% (p=0.0113). These postictal changes remained significant considering only complex partial seizures (n=17). The alterations were more pronounced in patients with hippocampal sclerosis. Treatment with valproic acid (VPA) was accompanied by a greater postictal decrease of CD4(+) T cells (25% compared to 5% in patients without VPA, p=0.041) while treatment with levetiracetam (LEV) correlated with a low postictal increase of NK-like T cells (4% versus 41%, p=0.016). Twenty-four hours after the seizures the alterations had resolved. CONCLUSION: Profound postictal changes in the immune cell composition of the peripheral blood may have been mediated by epinephrine release. The greater immune response in patients with mesial temporal lobe epilepsy due to hippocampal sclerosis may reflect a close relationship between mesial temporal structures and the sympathetic nerve system. VPA and LEV may have an impact on seizure induced immunological changes.