Items 1-20 of 4094

• Journal Article

#### A nanobody-based fluorescent reporter reveals human α-synuclein in the cell cytosol ﻿

Nature Communications 2020; 11(1) p.1-13: Art. 2729
Aggregation and spreading of α-Synuclein (αSyn) are hallmarks of several neurodegenerative diseases, thus monitoring human αSyn (hαSyn) in animal models or cell cultures is vital for the field. However, the detection of native hαSyn in such systems is challenging. We show that the nanobody NbSyn87, previously-described to bind hαSyn, also shows cross-reactivity for the proteasomal subunit Rpn10. As such, when the NbSyn87 is expressed in the absence of hαSyn, it is continuously degraded by the proteasome, while it is stabilized when it binds to hαSyn. Here, we exploit this feature to design a new Fluorescent Reporter for hαSyn (FluoReSyn) by fusing NbSyn87 to fluorescent proteins, which results in fluorescence signal fluctuations depending on the presence and amounts of intracellular hαSyn. We characterize this biosensor in cells and tissues to finally reveal the presence of transmittable αSyn in human cerebrospinal fluid, demonstrating the potential of FluoReSyn for clinical research and diagnostics.
• Journal Article

#### Longitudinal Case Study of Regression-Based Hand Prosthesis Control in Daily Life ﻿

Frontiers in Neuroscience 2020; 14 p.1-8: Art. 600
Hand prostheses are usually controlled by electromyographic (EMG) signals from the remnant muscles of the residual limb. Most prostheses used today are controlled with very simple techniques using only two EMG electrodes that allow to control a single prosthetic function at a time only. Recently, modern prosthesis controllers based on EMG classification, have become clinically available, which allow to directly access more functions, but still in a sequential manner only. We have recently shown in laboratory tests that a regression-based mapping from EMG signals into prosthetic control commands allows for a simultaneous activation of two functions and an independent control of their velocities with high reliability. Here we aimed to study how such regression-based control performs in daily life in a two-month case study. The performance is evaluated in functional tests and with a questionnaire at the beginning and the end of this phase and compared with the participant’s own prosthesis, controlled with a classical approach. Already 1 day after training of the regression model, the participant with transradial amputation outperformed the performance achieved with his own Michelangelo hand in two out of three functional metrics. No retraining of the model was required during the entire study duration. During the use of the system at home, the performance improved further and outperformed the conventional control in all three metrics. This study demonstrates that the high fidelity of linear regression-based prosthesis control is not restricted to a laboratory environment, but can be transferred to daily use.
• Journal Article

#### Compassionate Use of the ROCK Inhibitor Fasudil in Three Patients With Amyotrophic Lateral Sclerosis ﻿

Frontiers in Neurology 2020; 11 p.1-8: Art. 173
The Rho kinase (ROCK) inhibitor Fasudil is a promising drug for a disease-modifying therapy of amyotrophic lateral sclerosis (ALS). In preclinical models, Fasudil was shown to increase motor neuron survival, inhibit axonal degeneration, enhance axonal regeneration and modulate microglial function in vitro and in vivo. It prolonged survival and improved motor function of SOD1-G93A-mice. Recently, a phase IIa clinical trial has been commenced to investigate the safety, tolerability, and efficacy of Fasudil in ALS patients at an early stage of disease (ROCK-ALS trial, NCT03792490, Eudra-CT-Nr.: 2017-003676-31). Although Fasudil has been approved in Japan for many years for the treatment of vasospasms following subarachnoid hemorrhage and is known to have a favorable side effect profile in these patients, there is no data on its use in human patients with ALS or any other neurodegenerative conditions. Here, we report the first three cases of compassionate use of Fasudil in patients with ALS. Between May 2017 and February 2019, one male (66 years old) and two female (62 and 68 years old) subjects with probable or definite ALS according to the El Escorial criteria (one of the females having a pathogenic SOD1 mutation) were administered Fasudil 30 mg intravenously twice daily over 45 min on 20 consecutive working days. Blood pressure, heart rate and routine laboratory tests were constantly controlled. All three subjects tolerated the Fasudil infusions well without any obvious side effects. Interestingly, the slow vital capacity showed a significant increase in one of the patients. Taken together, we report here the first compassionate use of the ROCK inhibitor Fasudil in three ALS patients, which was well-tolerated.
• Journal Article

#### Prevention of Sexual Child Abuse: Preliminary Results From an Outpatient Therapy Program ﻿

Frontiers in Psychiatry 2020; 11 p.1-15: Art. 88
In Germany, access to outpatient treatment services devoted to the prevention of (further) sexual offenses against minors and child sexual exploitation material (CSEM) offenses is often limited. The therapy project “Prevention of Sexual Abuse” tries to fill this gap by providing treatment to patients with a self-reported sexual interest in children and adolescents, irrespective of whether or not they are pedophilic or prosecuted by the legal justice system. Within the project, a treatment manual was developed which specifically addresses dynamic risk-factors in child sexual abusers andCSEMoffenders. The treatment manualwas conceived to reduce recidivism risk and to contribute to the enhancement of the patients’ personal wellbeing. In this paper, results of the accompanying scientific research are presented: offensesupportive attitudes (N= 23), self-reportedCSEMuse (N= 10), emotional distress (N= 24), and participants’ subjective risk perception of committing (further) sexual offenses (N = 25) reduced during the course of treatment. A reduction of offense-supportive attitudes was further observed from pre-intervention to 1-year follow-up (N = 8). Changes with regard to selfefficacy, quality of life, participants’ self-perceived ability to control sexual impulses toward children and adolescents permanently, and several measures assessing different kinds of sexual recidivism did not, however, reach any level of significance. During an average observation period of 2.4 years, six patients confessed to have conducted new sexual exploitation material offenses, while no further sexual abuse cases were reported (N = 19). Due to the used research design and small sample sizes, treatment effects cannot be inferred and external validity is limited. This notwithstanding, results provide first evidence for a relationship between treatment participation and self-reported recidivism and psychological well-being.
• Journal Article

#### TraPS-VarI: Identifying genetic variants altering phosphotyrosine based signalling motifs ﻿

Scientific Reports 2020; 10(1) p.1-10: Art. 8453
Patient stratification and individualized therapeutic strategies rely on the established knowledge of genotype-specific molecular and cellular alterations of biological and therapeutic significance. Whilst almost all approved drugs have been developed based on the Reference Sequence protein database (RefSeq), the latest genome sequencing studies establish the substantial prevalence of non-synonymous genetic mutations in the general population, including stop-insertion and frame shift mutations within the coding regions of membrane proteins. While the availability of individual genotypes are becoming increasingly common, the biological and clinical interpretations of mutations among individual genomes is largely lagging behind. Lately, transmembrane proteins of haematopoietic (myeloid and lymphoid) derived immune cells have attracted much attention as important targets for cancer immunotherapies. As such, the signalling properties of haematological transmembrane receptors rely on the membrane-proximal phosphotyrosine based sequence motifs (TBSMs) such as ITAM (immunoreceptor tyrosine-based activation motif), ITIM (immunoreceptor tyrosine-based inhibition motif) and signal transducer and activator of transcription 3 (STAT3)-recruiting YxxQ motifs. However, mutations that alter the coding regions of transmembrane proteins, resulting in either insertion or deletion of crucial signal modulating TBSMs, remains unknown. To conveniently identify individual cell line-specific or patient-specific membrane protein altering mutations, we present the Transmembrane Protein Sequence Variant Identifier (TraPS-VarI). TraPS-VarI is an annotation tool for accurate mapping of the effect of an individual’s mutation in the transmembrane protein sequence, and to identify the prevalence of TBSMs. TraPS-VarI is a biologist and clinician-friendly algorithm with a web interface and an associated database browser (https://www.traps-vari.org/).
• Journal Article

#### Clinical Efficacy of Routinely Administered Belimumab on Proteinuria and Neuropsychiatric Lupus ﻿

Frontiers in Medicine 2020; 7 p.1-7: Art. 222
• Journal Article

#### Constructing temporal regulatory cascades in the context of development and cell differentiation ﻿

PLOS ONE 2020; 15(4) p.1-17: Art. e0231326
Cell differentiation is a complex process orchestrated by sets of regulators precisely appearing at certain time points, resulting in regulatory cascades that affect the expression of broader sets of genes, ending up in the formation of different tissues and organ parts. The identification of stage-specific master regulators and the mechanism by which they activate each other is a key to understanding and controlling differentiation, particularly in the fields of tissue regeneration and organoid engineering. Here we present a workflow that combines a comprehensive general regulatory network based on binding site predictions with user-provided temporal gene expression data, to generate a a temporally connected series of stage-specific regulatory networks, which we call a temporal regulatory cascade (TRC). A TRC identifies those regulators that are unique for each time point, resulting in a cascade that shows the emergence of these regulators and regulatory interactions across time. The model was implemented in the form of a user-friendly, visual web-tool, that requires no expert knowledge in programming or statistics, making it directly usable for life scientists. In addition to generating TRCs the tool links multiple interactive visual workflows, in which a user can track and investigate further different regulators, target genes, and interactions, directing the tool along the way into biologically sensible results based on the given dataset. We applied the TRC model on two different expression datasets, one based on experiments conducted on human induced pluripotent stem cells (hiPSCs) undergoing differentiation into mature cardiomyocytes and the other based on the differentiation of H1-derived human neuronal precursor cells. The model was successful in identifying previously known and new potential key regulators, in addition to the particular time points with which these regulators are associated, in cardiac and neural development.
• Journal Article

#### COVID-19: Possible Impact of the Genetic Background in $IFNL$ Genes on Disease Outcomes ﻿

Journal of Innate Immunity 2020; 12(3) p.273-274
• Journal Article

#### Loss of Hippocampal Calretinin and Parvalbumin Interneurons in the 5XFAD Mouse Model of Alzheimer’s Disease ﻿

ASN Neuro 2020; 12 p.1-12: Art. 1759091420925356
The deposition of amyloid-b peptides in the form of extracellular plaques and neuronal degeneration belong to the hallmark features of Alzheimer’s disease (AD). In addition, impaired calcium homeostasis and altered levels in calcium-binding proteins seem to be associated with the disease process. In this study, calretinin- (CR) and parvalbumin- (PV) positive gammaaminobutyric acid-producing (GABAergic) interneurons were quantified in different hippocampal subfields of 12-month-old wild-type mice, as well as in the transgenic AD mouse models 5XFAD and Tg4-42. While, in comparison with wild-type mice, CR-positive interneurons were mainly reduced in the CA1 and CA2/3 regions in plaque-bearing 5XFAD mice, PV-positive interneurons were reduced in all analyzed subfields including the dentate gyrus. No reduction in CR- and PV-positive interneuron numbers was detected in the non-plaque-forming Tg4-42 mouse, although this model has been previously demonstrated to harbor a massive loss of CA1 pyramidal neurons. These results provide information about hippocampal interneuron numbers in two relevant AD mouse models, suggesting that interneuron loss in this brain region may be related to extracellular amyloid burden.
• Journal Article

#### Urine E-cadherin: A Marker for Early Detection of Kidney Injury in Diabetic Patients ﻿

Journal of Clinical Medicine 2020; 9(3) p.1-21: Art. 639
Diabetic nephropathy (DN) is themain reason for end-stage renal disease. Microalbuminuria as the non-invasive available diagnosismarker lacks specificity and gives high false positive rates. To identify and validate biomarkers for DN, we used in the present study urine samples from four patient groups: diabetes without nephropathy, diabetes withmicroalbuminuria, diabetes withmacroalbuminuria and proteinuria without diabetes. For the longitudinal validation, we recruited 563 diabetic patients and collected 1363 urine samples with the clinical data during a follow-up of 6 years. Comparative urinary proteomics identified four proteins Apolipoprotein A-I (APOA1), Beta-2-microglobulin (B2M), E-cadherin (CDH1) and Lithostathine-1-alpha (REG1A), which differentiated with high statistical strength ($p$ < 0.05) between DNpatients and the other groups. Label-freemass spectrometric quantification of the candidates confirmed the discriminatory value of E-cadherin and Lithostathine-1-alpha ($p$ < 0.05). Immunological validation highlighted E-cadherin as the only marker able to differentiate significantly between the different DN stages with an area under the curve (AUC) of 0.85 (95%-CI: [0.72, 0.97]). The analysis of the samples from the longitudinal study confirmed the prognostic value of E-cadherin, the critical increase in urinary E-cadherin level was measured 20 $\pm$ 12.5 months before the onset of microalbuminuria and correlated significantly ($p$ < 0.05) with the glomerular filtration rate measured by estimated glomerular filtration rate (eGFR).
• Journal Article

#### The role of cellular prion protein in lipid metabolism in the liver ﻿

Prion 2020; 14(1) p.95-108
Cellular prion protein (PrPC) is a plasma membrane glycophosphatidylinositol-anchored proteinand it is involved in multiple functions, including neuroprotection and oxidative stress. So far,most of the PrPC functional research is done in neuronal tissue or cell lines; the role of PrPC innon-neuronal tissues such as liver is only poorly understood. To characterize the role of PrPC inthe liver, a proteomics approach was applied in the liver tissue of PrPC knockout mice. Theproteome analysis and biochemical validations showed an excessive fat accumulation in the liverof PrPC knockout mice with a change in mRNA expression of genes linked to lipid metabolism. Inaddition, the higher Bax to Bcl2 ratio, up-regulation of tgfb1 mRNA expression in PrPC knockoutmice liver, further showed the evidences of metabolic disease. Over-expression of PrPC in fattyacid-treated AML12 hepatic cell line caused a reduction in excessive intracellular fat accumulation;shows association of PrPC levels and lipid metabolism. Therefore, based on observation ofexcessive fat globules in the liver of ageing PrPC knockout mice and the reduction of fataccumulation in AML12 cell line with PrPC over-expression, the role of PrPC in lipid metabolismis described.
• Journal Article

#### Zebrafish disease model of human RNASET2-deficient cystic leukoencephalopathy displays abnormalities in early microglia ﻿

Biology Open 2020; 9(5) p.1-10: Art. bio049239
Human infantile-onset RNASET2-deficient cystic leukoencephalopathy is a Mendelian mimic of in utero cytomegalovirus brain infection with prenatally developing inflammatory brain lesions. We used an RNASET2-deficient zebrafish model to elucidate the underlying disease mechanisms. Mutant and wild-type zebrafish larvae brain development between 2 and 5 days post fertilization (dpf) was examined by confocal live imaging in fluorescent reporter lines of the major types of brain cells. In contrast to wild-type brains, RNASET2-deficient larvae displayed increased numbers of microglia with altered morphology, often containing inclusions of neurons. Furthermore, lysosomes within distinct populations of the myeloid cell lineage including microglia showed increased lysosomal staining. Neurons and oligodendrocyte precursor cells remained unaffected. This study provides a first look into the prenatal onset pathomechanisms of human RNASET2-deficient leukoencephalopathy, linking this inborn lysosomal disease to the innate immune system and other immune-related childhood encephalopathies like Aicardi-Goutières syndrome (AGS).
• Journal Article

#### German and Italian Users of Web- Accessed Genetic Data: Attitudes on Personal Utility and Personal Sharing Preferences. Results of a Comparative Survey (n=192) ﻿

Frontiers in Genetics 2020; 11 p.1-13: Art. 102
Genetic information is increasingly provided outside of the traditional clinical setting, allowing users to access it directly via specialized online platforms. This development is possibly resulting in changing ethical and social challenges for users of predictive genetic tests. Little is known about the attitudes and experiences of users of web-accessed genetic information. This survey analyzes data from two European countries with regard to the utility of genetic information, the users’ ways of making use of and dealing with information, and their sharing behavior. Particular focus is given to ethical and social questions regarding the motivation to share personal genetic results with others. Social factors tested for are national background, gender, and marital, parental, and educational status. This study will contribute to public discourse and offer ethical recommendations. The study will also serve to validate the developed questionnaire for use in population representative surveys.
• Journal Article

#### Shedding New Light on Cancer Metabolism: A Metabolic Tightrope Between Life and Death ﻿

Frontiers in Oncology 2020; 10 p.1-18: Art. 409
Since the earliest findings of Otto Warburg, who discovered the first metabolic differences between lactate production of cancer cells and non-malignant tissues in the 1920s, much time has passed. He explained the increased lactate levels with dysfunctional mitochondria and aerobic glycolysis despite adequate oxygenation. Meanwhile, we came to know that mitochondria remain instead functional in cancer cells; hence, metabolic drift, rather than being linked to dysfunctional mitochondria, was found to be an active act of direct response of cancer cells to cell proliferation and survival signals. This metabolic drift begins with the use of sugars and the full oxidative phosphorylation via the mitochondrial respiratory chain to form CO$_2$, and it then leads to the formation of lactic acid via partial oxidation. In addition to oncogene-driven metabolic reprogramming, the oncometabolites themselves alter cell signaling and are responsible for differentiation and metastasis of cancer cells. The aberrant metabolism is now considered a major characteristic of cancer within the past 15 years. However, the proliferating anabolic growth of a tumor and its spread to distal sites of the body is not explainable by altered glucose metabolism alone. Since a tumor consists of malignant cells and its tumor microenvironment, it was important for us to understand the bilateral interactions between the primary tumor and its microenvironment and the processes underlying its successful metastasis. We here describe the main metabolic pathways and their implications in tumor progression and metastasis. We also portray that metabolic flexibility determines the fate of the cancer cell and ultimately the patient. This flexibility must be taken into account when deciding on a therapy, since singular cancer therapies only shift the metabolism to a different alternative path and create resistance to the medication used. As with Otto Warburg in his days, we primarily focused on the metabolism of mitochondria when dealing with this scientific question.
• Journal Article

#### UDP-glucose 6-dehydrogenase expression as a predictor of survival in patients with pulmonary adenocarcinoma ﻿

International Journal of Surgery Oncology 2020; 5(2) p.1-5: Art. e85
Background: UDP-glucose-6-dehydrogenase (UGDH) plays an important role in the production of hyaluronic acid, an extracellular matrix component that is responsible for the promotion of normal cellular growth and migration. Increased levels of UGDH have been linked to the progression of epithelial cancers, such as those of the breast, colon and prostate. Therefore we aimed to analyze if the expression level of UGDH does also influence patients survival of lung cancer patients. Methods: UGDH expression levels were analyzed by immunohistochemistry in 96 samples of pulmonary adenocarcinoma (AC), 84 cases of squamous cell lung carcinoma (SQCLC) and 33 samples of small cell lung cancer (SCLC) and correlated with clinicopathologic characteristics and patient outcome. Results: UGDH was expressed in 62.5% cases of AC, 70.2% cases of SQCLC, and 48.5% cases of SCLC. In AC, expression of UGDH was significantly associated with lymph node metastasis and worse overall survival of the affected patients. However, UGDH expression had no significant correlation to prognosis in SQCLC or SCLC patients. Conclusions: In our study, expression of UGDH was associated with worse prognosis of patients with pulmonary adenocarcinoma so that expression of UGDH might help to guide treatment decisions. Furthermore, UGDH might present a potential novel drug target in AC as it displays inhibitable catalytic activity.
• Journal Article

#### Silver linings on the horizon: highlights from the 10th Cachexia Conference ﻿

Journal of Cachexia, Sarcopenia and Muscle 2018; 9(1) p.176-182
This article highlights the updates from preclinical and clinical studies into the field of wasting disorders that were presented at the 10th Cachexia Conference held in Rome, Italy, in December 2017. This year’s conference saw some interesting results of larger-scale studies and clinical trials and new therapeutic targets. Herein, we summarize the biological and clinical significance of different markers and new diagnostic tools and cut-offs for the detection of skeletal muscle wasting, including micro RNAs, the ubiquitin-proteasome system, mTOR signalling, news in body composition analysis including the D3-creatine dilution method, and new biomarkers. Clinical studies investigated novel nutritional approaches, trials of elamipretide, enobosarm, and urolithin A. It remains a fact, however, that effective treatments of cachexia and wasting disorders are urgently needed in order to improve patients’ quality of life and their survival.
• Journal Article

#### Factorial Structure and Validity of Depression (PHQ-9) and Anxiety (GAD-7) Scales after Traumatic Brain Injury ﻿

Journal of Clinical Medicine 2020; 9(3) p.1-21: Art. 873
Background: The dimensionality of depression and anxiety instruments have recently been a source of controversy. Objectives and Design: in a European-wide sample of patients after Traumatic Brain Injury (TBI), we aim to examine the factorial structure, validity, and association of the Patient Health Questionnaire for depression (PHQ-9) and the Generalized Anxiety Disorder (GAD-7) instruments. This study is based on longitudinal observational data. We conducted analyses of factorial structure and discriminant validity of outcomes six-months after TBI. We also examined the prevalence, co-occurrence, and changes of scores on the PHQ-9 and GAD-7 at 3-, 6-, and 12-month post-TBI assessments. Participants: At six-months post-TBI assessment, 2137 (738 (34.5%) women) participants completed the PHQ-9 and GAD-7 questionnaires. For the longitudinal analysis, we had 1922 participants (672 (35.0%) women). Results: The results of exploratory factor analysis suggested a general latent construct underlying both PHQ-9 and GAD-7 measures. Confirmatory factor analyses showed a slight improvement in the fit indices for the bifactorial model. The Omega hierarchical test clearly di erentiated two subfactors of PHQ-9 and GAD-7 items over and above the underlying general factor; however, most of the variance (85.0%) was explained by the general factor and the explained variance of the subfactors was small. The PHQ-9 and GAD-7 performed similarly in detecting post-traumatic stress disorder (PTSD). As defined by conventional cut-o s, depression and anxiety have di erent prevalence rates in the sample. The scales also di ered in their relationships with the short form of health survey (SF-36v2) subscales. The longitudinal analysis showed high stability of depression and anxiety symptoms: 49–67% of the post-TBI patients with comorbid depression and anxiety reported the persistence of the symptoms over time. Discussion: The factorial structure analysis favors a general latent construct underlying both depression and anxiety scales among patients after TBI. We discuss the implications our findings and future research directions.
• Journal Article

#### Discriminating malignant and benign clinical T1 renal masses on computed tomography ﻿

Medicine 2020; 99(16) p.1-8: Art. e19725
The aim of this study was to discriminate malignant and benign clinical T1 renal masses on routinely acquired computed tomography (CT) images using radiomics and machine learning techniques. Adult patients undergoing surgical resection and histopathological analysis of clinical T1 renal masses were included. Preoperative CT studies in venous phase from multiple referring centers were included, without restriction to specific CT scanners, slice thickness, or degrees of artifacts. Renalmasseswere segmented and 120 standardized radiomic features extracted. Machine learning algorithms were used to predictmalignancy of renal masses using radiomics features and cross-validation. Diagnostic accuracy of machine learningmodels and assessment by independent blinded radiologists were compared based on the gold standard of histopathologic diagnosis. A total of 94 patients met inclusion criteria (benign renal masses: n=18; malignant: n=76). CT studies from 18 different scanners were assessed with median slice thickness of 2.5mm and artifacts in 15 cases (15.9%). Area under the receiver-operating-characteristics curve (AUC) of random forest (random forest [RF], AUC=0.83) was significantly higher compared to the radiologists (AUC=0.68, P=.047). Sensitivity was significantly higher for RF versus radiologists (0.88 vs 0.80, P=.045), whereas specificity was numerically higher for RF (0.67 vs 0.50, P=.083). Although limited by an overall small sample size and few benign renal tumors, a radiomic features and machine learning approach suggests a high diagnostic accuracy for discrimination of malignant and benign clinical T1 renal masses on venous phase CT. The presented algorithm robustly outperforms human readers in a real-life scenario with nonstandardized imaging studies from various referring centers.
• Journal Article

#### Significant effusion in the joints of the lower extremity after running an ultramarathon in extreme conditions ﻿

Journal of Human Sport and Exercise 2020; 15(4)
Introduction: The “Brocken challenge” ultramarathon takes place in the cold of February over 80 km with 1,900 m of elevation change. The purpose of this study was to evaluate the effects of an ultramarathon under extreme conditions on the lower extremities. Methods: Out of the 182 starters, 44 athletes were included into the study (n=44). We examined these athletes using a questionnaire, by measuring circumferences of their lower extremities and by standardized sonographic measurement of joint effusion of knee and ankle joints before and after the run. Results: After the run, the right leg and both feet significantly increased in circumference. Knee joints on both sides and the left ankle joint showed significantly more effusion after the run (right knee: 84%, right ankle: 43%; left knee: 80%, left ankle: 48%). Heavier and less trained athletes showed significantly more effusion in sonographic assessment of the knee and ankle. Neither swelling, nor effusion had a measurable influence on finishing time. Conclusions: Running an ultramarathon in the cold overloads the fluid draining capacity in the muscles and joints of the legs especially in heavier and less trained athletes without measurable immediate effect on performance.
• Journal Article

#### Judgments of effort exerted by others are influenced by received rewards ﻿

Scientific Reports 2020; 10(1) p.1-14: Art. 1868